3D Echocardiography Provides Highly Accurate 3D Printed Models in Congenital Heart Disease.

Cardiac 3D printing is mainly performed from magnetic resonance imaging (MRI) and computed tomography (CT) 3D datasets, though anatomic detail of atrioventricular (AV) valves may be limited. 3D echo provides excellent visualization of AV valves. Thus, we tested the feasibility and accuracy of 3D printing from 3D echo in this pilot series of subjects with congenital heart disease (CHD), with a focus on valve anatomy. Five subjects with CHD were identified. 3D echo data were converted to 3D printable files and printed in collaboration with 3D Systems Healthcare (Golden, Colorado). A novel technique for valve modeling was utilized using commercially available software. Two readers (KM, SA) independently measured valve structures from 3D models and compared to source echo images. 3D printing was feasible for all cases. Table 1 shows measurements comparing 2D echo to 3D models. Bland Altman analysis showed close agreement and no significant bias between 2D and digital 3D models (mean difference 0.0, 95% CI 1.1 to - 1.1) or 2D vs printed 3D models, though with wider limits of agreement (mean difference - 0.3, 95% CI 1.9 to - 2.6). Accuracy of 3D models compared to 2D was within < 0.5 mm. This pilot study shows 3D echo datasets can be used to reliably print AV and semilunar valve structures in CHD. The 3D models are highly accurate compared to the source echo images. This is a novel and value-added technique that adds incremental information on cardiac anatomy over current methods.

Striatal dopamine D1-type receptor availability: no difference from control but association with cortical thickness in methamphetamine users.

Chronic methamphetamine use poses potentially devastating consequences for directly affected individuals and for society. Lower dopamine D2-type receptor availability has been observed in striata of methamphetamine users as compared with controls, but an analogous comparison of D1-type receptors has been conducted only on post-mortem material, with no differences in methamphetamine users from controls in the caudate nucleus and putamen and higher D1-receptor density in the nucleus accumbens. Released from neurons when methamphetamine is self-administered, dopamine binds to both D1- and D2-type receptors in the striatum, with downstream effects on cortical activity. Thus, both receptor subtypes may contribute to methamphetamine-induced alterations in cortical morphology and behavior. In this study, 21 methamphetamine-dependent subjects and 23 healthy controls participated in positron emission tomography and structural magnetic resonance imaging for assessment of striatal D1- and D2-type receptor availability and cortical gray-matter thickness, respectively. Although D2-type receptor availability (BPnd) was lower in the methamphetamine group, as shown previously, the groups did not differ in D1-type BPnd. In the methamphetamine group, mean cortical gray-matter thickness was negatively associated with cumulative methamphetamine use and craving for the drug. Striatal D1-type but not D2-type BPnd was negatively associated with global mean cortical gray-matter thickness in the methamphetamine group, but no association was found between gray-matter thickness and BPnd for either dopamine receptor subtype in the control group. These results suggest a role of striatal D1-type receptors in cortical adaptation to chronic methamphetamine use.

Cerebrovascular reactivity after cessation of menopausal hormone treatment.

OBJECTIVE: Women who are currently using menopausal hormone therapy (MHT) have higher cerebrovascular reactivity when compared with postmenopausal women who are not taking MHT; however, the effect of cessation of MHT on cerebrovascular reactivity is not known. Given that MHT can have structural and activational effects on vascular function, this study was performed to characterize cerebrovascular reactivity following cessation of MHT in women at low risk for cerebrovascular disease. METHODS: Cerebrovascular reactivity was measured in a subset of women from the Kronos Early Estrogen Prevention Study (KEEPS) 3 years after cessation of the study drug (oral conjugated equine estrogen, transdermal 17ß-estradiol, or placebo [PLA]). RESULTS: Age, body mass index, and blood pressure were comparable among groups. At rest, the middle cerebral artery velocity (MCAv), cerebrovascular conductance index, mean arterial pressure, and cerebral pulsatility index did not differ among groups. Slope-based summary measures of cerebrovascular reactivity did not differ significantly among groups. However, utilizing repeated-measures modeling, there was a significant upward shift in MCAv responses (p = 0.029) in the combined MHT group compared with the PLA group. CONCLUSION: MHT has a marginal sustained effect on cerebrovascular reactivity when measured 3 years after cessation of hormone treatment.

Inhibiting c-Jun N-terminal kinase partially attenuates caffeine-dependent cell death without alleviating the caffeine-induced reduction in mitochondrial respiration in C2C12 skeletal myotubes.

Caffeine is a widely consumed stimulant that has previously been shown to promote cytotoxic stress and even cell death in numerous mammalian cell lines. Thus far there is little information available regarding the toxicity of caffeine in skeletal muscle cells. Our preliminary data revealed that treating C2C12 myotubes with 5 mM caffeine for 6 h increased nuclear fragmentation and reduced basal and maximal oxygen consumption rate (OCR) in skeletal myotubes. The purpose of this study was to further elucidate the pathways by which caffeine increased cell death and reduced mitochondrial respiration. We specifically examined the role of c-Jun N-terminal kinase (JNK), which has previously been shown to simultaneously increase caspase-dependent cell death and reduce mitochondrial respiration in other mammalian cell lines. We found that caffeine promoted a dose-dependent increase in cell death in multinucleated myotubes but did not in mononucleated myoblasts. The addition of 10 µM Z-DEVD-FMK, a specific inhibitor of executioner caspases, completely inhibited caffeine-dependent cell death. Further, the addition of 400 µM dantrolene, a specific ryanodine receptor (RYR) inhibitor, prevented the caffeine-dependent increase in cell death and the reduction in basal and maximal OCR. We also discovered that caffeine treatment significantly increased the phosphorylation of JNK and that the addition of 30 µM SP600125 (JNKi), a specific JNK inhibitor, partially attenuated caffeine-induced cell death without preventing the caffeine-dependent reduction in basal and maximal OCR. Our results suggest that JNK partially mediates the increase in caspase-dependent cell death but does not contribute to reduced mitochondrial respiration in caffeine-treated skeletal muscle cells. We conclude that caffeine increased cell death and reduced mitochondrial respiration in a calcium-dependent manner by activating the RYR and promoting reticular calcium release.

Two-dimensional crystallization conditions of human leukotriene C4 synthase requiring adjustment of a particularly large combination of specific parameters.

Human leukotriene C(4) synthase (LTC(4)S) forms highly ordered two-dimensional (2D) crystals under specific reconstitution conditions. It was found that control of a larger number of parameters than is usually observed for 2D crystallization of membrane proteins was necessary to induce crystal formation of LTC(4)S. Here, we describe the parameters that were optimized to yield large and well-ordered 2D crystals of LTC(4)S. Careful fractioning of eluates during the protein purification was essential for obtaining crystals. While the lipid-to-protein ratio was critical in obtaining order, four parameters were decisive in inducing growth of crystals that were up to several microns in size. To obtain a favorable diameter, salt, temperature, glycerol, and initial detergent concentration had to be controlled with great care. Interestingly, several crystal forms could be grown, namely the plane group symmetries of p2, p3, p312, and two different unit cell sizes of plane group symmetry p321.

Cloning and sequencing of a cDNA expressing a recombinant house dust mite protein that binds human IgE and corresponds to an important low molecular weight allergen.

A cDNA clone coding for a mite allergen of mol wt approximately 14,000 has been isolated and its DNA sequence determined. The native component from mite extracts encoded by this DNA was identified by immunoprobing blots of mite body extract with human IgE eluted from the electroblotted cloned fusion polypeptides derived from the expressed cDNA clone. The clone encodes a polypeptide with a deduced mol wt of 17,460. The deduced amino acid sequence was not homologous to any known protein sequences and it contains no cysteine or tryptophan. On blots, 21 of 38 sera from mite-allergic subjects recognized the cloned material, and this recognition strongly correlated with IgE-binding to the native component on protein blots of mite extract.

Association between MMP1 and MMP9 activities and ICAM1 cleavage induced by tumor necrosis factor in stromal cell cultures from eutopic endometria of women with endometriosis.

Endometriosis is a benign gynecological pathology in which immune system deregulation may play a role in its initiation and progression. In endometriotic lesions, intercellular adhesion molecule-1 (ICAM1) is released from the cell membrane by proteolytic cleavage of its extracellular domain, a process that coincides with increased expression and proteolytic activity of metalloproteinases such as MMP1 and MMP9. The objective of our study was to investigate the association between MMP1 and MMP9 activities and ICAM1 cleavage mediated by tumor necrosis factor (TNF) in eutopic endometrial stromal cells from women with and without (control) endometriosis during culture. The RNA was evaluated by RT-PCR, and the protein was determined by western blot (ICAM1, MMP1), casein or gelatin zymographies (secreted active MMP1 or MMP9 respectively), ELISA (soluble ICAM1 (sICAM1)), and fluorescence assay (secreted active MMP1). Under basal conditions, proMMP9 dimer and MMP9 were higher in endometriosis cell cultures. In stromal cultures derived from control women and those with endometriosis, TNF augmented the intracellular proMMP1 (1.2-fold in control stromal cells) and ICAM1 (1.4- and 1.9-fold), greatly increased MMP1 and proMMP9 levels, and the sICAM1 concentration (2.3- and 4.3-fold) in their media compared with basal levels. The combination of TNF and MMP9 increased the sICAM1 concentration 14-fold in the endometriosis cell media, whereas GM6001 inhibited the stimulatory effect of TNF in both cell cultures. The deregulation of MMP9, and the TNF participation in the MMP1 and proMMP9 secretions, in the MMP9 expression and in the expression and cleavage of ICAM1 may contribute to the pathophysiology of this disease.

Extremely rapid visual search: the maximum rate of scanning letters for the presence of a numeral.

Subjects searched a rapid sequence of computer-produced letter arrays for the presence of a numeral in one of the arrays. The subjects' scanning rates were computed from their precentage of correct detections of the location of the numeral. Scanning rates were very high and approximately the same for a wide variety of conditions; the highest scanning rates (125 and 75 letters per second for two subjects) occurred when there were 9 or 16 letters in each of the arrays and when new arrays were presented every 40 to 50 milliseconds. Giving the subject advance knowledge of the numeral to be presented made little difference in the scores.

Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms.

Ionophores and supplemental fat are fed to lactating cows to improve feed efficiency. Their effect on rumen fermentation is similar, but less is known about their impact on rumen microbes. The objective of this study was to determine the effects of monensin (M), bacitracin (B), and soybean oil (O) on microbial populations. Mixed cultures of rumen microbes were incubated in 5 dual-flow continuous fermentors and fed 13.8 g of alfalfa hay pellets daily (DM basis) for 16 d. All fermentors were allowed to stabilize for 4 d. From d 5 to 10, two fermentors received O (5% of diet DM), one fermentor received M (22 mg/kg), and one received B (22 mg/kg). From d 11 to 16, the 2 fermentors receiving O also received either M (OM) or B (OB) and O was included in the fermentors receiving M (MO) and B (BO). One fermentor served as the control and received 100% alfalfa pellets throughout the experiment. Each run was replicated 3 times. Samples were taken at 2 h after the morning feeding on d 4, 10, and 16 and were analyzed for bacterial populations using terminal restriction fragment length polymorphism. Volatile fatty acid concentration, methane production, and pH in the control cultures were not affected by time and remained similar during the entire experiment. The M and O treatments reduced molar concentration of acetate, increased concentration of propionate, and decreased methane production. Bacitracin did not alter acetate or propionate concentration, but reduced methane production. All 3 treatments (M, B, and O) altered the fragment patterns of microbial profiles. In contrast, treatments MO, OM, BO, and OB had little effect on culture fermentation despite differences in the patterns of microbial fragments. The terminal restriction fragment length polymorphism data suggest that microbial adaptation to the in vitro system in the control fermentor occurred within 4 d.

Synthetic routes to a coordinatively unsaturated ruthenium complex supported by a tripodal, protic bis(N-heterocyclic carbene) phosphine ligand.

A facile, one pot synthesis of a coordinatively unsaturated ruthenium complex supported by a tripodal, protic bis(N-heterocyclic carbene) phosphine ligand is presented. A number of coordination complexes were discovered en route during this synthesis, revealing some of the unique aspects of complexes ligated by this type of tridentate, protic bis(NHC) ligand. Through a combination of 1D and 2D NMR spectroscopic analysis and single crystal X-ray diffraction, we reveal the intermediacy of phosphine-ligated bisimidazole complexes and show that abstraction of inner-sphere halide ions facilitates conversion to the desired tridentate bis(NHC) coordination mode. Ultimately the use of N-methyl-2-pyrrolidone is shown to enable the use of the extreme temperatures needed to facilitate the direct, thermally activated tautomerization reaction that gives rise to the bis(NHC) motif.

Mutations in the cardiac transcription factor NKX2.5 affect diverse cardiac developmental pathways.

Heterozygous mutations in NKX2.5, a homeobox transcription factor, were reported to cause secundum atrial septal defects and result in atrioventricular (AV) conduction block during postnatal life. To further characterize the role of NKX2.5 in cardiac morphogenesis, we sought additional mutations in groups of probands with cardiac anomalies and first-degree AV block, idiopathic AV block, or tetralogy of Fallot. We identified 7 novel mutations by sequence analysis of the NKX2.5-coding region in 26 individuals. Associated phenotypes included AV block, which was the primary manifestation of cardiac disease in nearly a quarter of affected individuals, as well as atrial septal defect and ventricular septal defect. Ventricular septal defect was associated with tetralogy of Fallot or double-outlet right ventricle in 3 individuals. Ebstein's anomaly and other tricuspid valve abnormalities were also present. Mutations in human NKX2.5 cause a variety of cardiac anomalies and may account for a clinically significant portion of tetralogy of Fallot and idiopathic AV block. The coinheritance of NKX2.5 mutations with various congenital heart defects suggests that this transcription factor contributes to diverse cardiac developmental pathways.

Circadian distribution of serum cytokines in multiple sclerosis.

AIMS: The purpose of this study was to examine circadian distribution of selected cytokine levels (IL-2, IL-10, GM-CSF, TNF-alpha, and IFN-gamma) in serum of subjects with active Multiple Sclerosis (MS) and non-MS subjects. MATERIALS AND METHODS: Six females (36-56y) and five males (52-68y) with active MS volunteered and consented for the study conducted at Special Diagnostic Ward of this hospital. All subjects gave their medical history and were given complete physical examination. Low purine meals were served at 16:30, 07:30 and 13:00 h. Lights were "OFF' at 22:30 hr and "ON" at 06:30h. Blood collections were made at 3h intervals over a 24h period of time. Six healthy male subjects (53-76y) subjects' data were obtained from a study conducted 3 years previously using the same procedural protocol. Cytokine assays were assessed using commercial enzyme-linked immuno-absorbent procedure. Time series of average data and the range of change between the highest and lowest concentrations are presented for MS subjects along with data from non-MS subjects. RESULTS: IL-2, IL-10, and GM-CSF levels were significantly reduced in females with MS when compared with levels of healthy subjects while their IL-6 levels were increased. The IL-6, GM-CSF and TNF-alpha levels in males with MS were below detection limits. The TNF-alpha levels were essentially similar in MS females and healthy subjects. CONCLUSIONS: These preliminary studies, although with very small number of patients and healthy male controls appear to suggest that the circadian analysis of cytokines and other markers of immunity may have utility in understanding the pathogenesis of diseases like MS.

[Elbow dislocation]. / Luxación de codo.

Elbow dislocation is the most frequent dislocation in the upper limb after shoulder dislocation. Closed reduction is feasible in outpatient care when there is no associated fracture. A review is presented of the different reduction procedures.

Biochemical evaluation of storage lesion in canine packed erythrocytes.

OBJECTIVES: To describe the biochemical changes - also known as the storage lesion - that occur in canine packed red blood cells during ex vivo storage. MATERIALS AND METHODS: Ten 125-mL units of non-leuco-reduced packed red blood cells in citrate phosphate dextrose adenine were obtained from a commercial blood bank within 24 hours of donation. Samples were aseptically collected on days 1, 4, 7, 14, 28, 35 and 42 for measurement of sodium, potassium, chloride, lactate, glucose, pH and ammonia concentrations. All units were cultured on day 42. Friedman's repeated measures test with Dunn's multiple comparison test was used for non-parametric data. A repeated-measures analysis of variance with Tukey's multiple comparison test was used for parametric data. Alpha was set to 0·05. RESULTS: All analytes changed significantly during storage. The mean ammonia on day 1 (58·14 g/dL) was significantly lower (P<0·05) than those on days 28 (1266 g/dL), 35 (1668 g/dL) and 42 (1860 g/dL). A significant increase in median lactate concentration over time was also observed, with day 1 (4·385 mmol/L) being significantly less (P<0·05) than days 14 (19·82 mmol/L), 21 (22·81 mmol/L), 35 (20·31 mmol/L) and 42 (20·81 mmol/L). Median pH was significantly decreased after day 7. All bacterial cultures were negative. CLINICAL SIGNIFICANCE: Many biochemical alterations occur in stored canine packed red blood cells, although further studies are required to determine their clinical importance.

Biochemical evaluation of the effects of storage on feline erythrocytes.

OBJECTIVE: To describe the biochemical changes that occur during storage of feline packed red blood cells. METHODS: Feline packed red blood cells were obtained from the manufacturer via overnight delivery immediately following collection. Bag spikes were placed using aseptic technique and samples were drawn on days 1, 4, 7, 14, 21, 28 and 35. Sodium, potassium, chloride, glucose, lactate, pH and ammonia were measured at each time point. Aerobic and anaerobic bacterial cultures were submitted following collection on day 35. RESULTS: There were statistically significant increases in the median concentrations of lactate and ammonia within the first 2 weeks of storage to a concentration of 12·38 mmol/L and 447·96 µmol/L, respectively. Glucose concentrations decreased significantly by day 28 to a mean of 1·86 mmol/L. Median sodium and chloride concentrations increased throughout the course of storage to a concentration of 158·20 and 131·00 mmol/L, respectively. Mean potassium concentrations decreased to a concentration of 2·40 mmol/L. CLINICAL SIGNIFICANCE: These results show that biochemical derangements within feline packed red blood cells are progressive, with some alterations, such as lactate and ammonia, occurring early within the storage periods, while others, including glucose and electrolytes, are slower to develop. Additional prospective research evaluating the clinical effects of these biochemical alterations is required.

Time is the enemy: Mortality in trauma patients with hemorrhage from torso injury occurs long before the "golden hour".

INTRODUCTION: The concept of the "Golden Hour" has been a time-honored tenet of prehospital trauma care, despite a paucity of data to substantiate its validity. Non-compressible torso hemorrhage has been demonstrated to be a significant cause of mortality in both military and civilian settings. We sought to characterize the impact of prehospital time and torso injury severity on survival. Furthermore, we hypothesized that time would be a significant determinant of mortality in patients with higher Abbreviated Injury Scale (AIS) grades of torso injury (AIS ≥ 4) and field hypotension (prehospital SBP ≤ 110 mmHg) as these injuries are commonly associated with hemorrhage. METHODS: Data for this analysis was generated from a registry of 2,523,394 injured patients entered into the National Trauma Data Bank Research Data Set from 2012 to 2014. Patients with torso injury were identified utilizing Abbreviated Injury Scale (AIS) for body regions 4 (Thorax) and 5 (Abdomen). Specific inclusion criteria for this study included pre-hospital time, prehospital SBP ≤110 mmHg, torso injury qualified by AIS and mortality. Patients with non-survivable torso injury (AIS = 6), severe head injuries (AIS ≥ 3), no signs of life in the field (SBP = 0), interfacility transfers, or those with any missing data elements were excluded. This classification methodology identified a composite cohort of 42,135 adult patients for analysis. RESULTS: The overall mortality rate of the study population was 7.9% (3326/42,135); Torso AIS and prehospital time were noted to be strong independent predictors of patient mortality in all population strata of the analysis (P < 0.05). The data demonstrated a profound incremental increase in mortality in the early time course after injury associated with torso AIS ≥4. CONCLUSION: In patients with high-grade torso injury, AIS grades ≥4, the degree anatomic disruption is associated with significant hemorrhage. In our study, a precipitous rise in patient mortality was exhibited in this high-grade injury group at prehospital times <30 min. Our data highlight the critical nature of prehospital time in patients with non-compressible torso hemorrhage. However, realizing that evacuation times ≤30 min may not be realistic, particularly in rural or austere environments, future efforts should be directed toward the development of therapies to increase the window of survival in the prehospital environment.

Resolution of right heart enlargement after closure of secundum atrial septal defect with transcatheter technique.

OBJECTIVES: The purpose of this study was to prospectively characterize the reduction in right atrial (RA) area and right ventricular (RV) volume after transcatheter closure of atrial septal defect (ASD) and to investigate factors that may predict magnitude of resolution in right heart enlargement. BACKGROUND: Secundum ASD can cause volume overload of the right side of the heart with the potential for development of late complications. Little is known about reduction in right heart size after closure of ASD. METHODS: Transthoracic echocardiography was performed in 38 patients undergoing transcatheter closure of ASD. The RA area and RV volume were measured prior (n = 38), within 24 hours (n = 37), at 3 to 6 months (n = 24), at 12 months (n = 20) and at 24 months (n = 10) after closure of ASD. Change over time within the study group was assessed and the study group was compared to a control group of 19 patients with structurally normal hearts. RESULTS: Indexed RA area decreased from baseline to 3- to 6-month follow-up (p = 0.004) as did indexed RV volume (p < 0.0001). Indexed RV volume was similar to that in the control group at 24 months (p = 0.3); however, indexed RA area remained greater than in the control group (p = 0.006). Decrease in indexed RA area over the first 12 months of follow-up was related to young age at time of closure by regression analysis (r = 0.55, p = 0.013). CONCLUSION: Closure of secundum ASD results in decreased indexed RV volume comparable to that in control subjects at 24 months following closure. Indexed RA area remains increased compared to that in control subjects but does decrease over time. Decrease in RA area is inversely proportional to age at time of ASD closure. Long-term follow-up is required to evaluate the clinical impact of persistently increased RA size.

Determination of Anthocyanins and Total Polyphenols in a Variety of Elderberry Juices by UPLC-MS/MS and Other Methods.

Elderberry (Sambucus spp.) juice contains a variety of polyphenols, mostly anthocyanins. In order to understand the variation of polyphenol levels by genotype, various elderberry juice samples were analyzed for total phenolics (TP), total monomeric anthocyanins (TMA) and individual anthocyanin content. The Folin-Ciocalteu total phenolic method and pH differential method were used to measure the TP and TMA content, respectively. The TP and TMA concentrations of elderberry were found to vary greatly among different genotypes. TMA content varied from 2.1% for 'Sperandio' to 60.6% for the 'Bob Gordon' cultivar. In addition, ultra-performance liquid chromatography with triple quadrupole mass spectrometry was used to separate and detect individual anthocyanins from samples prepared by solid phase extraction. Multiple-reaction-monitoring was used to process data for the reduction of false positives, maximizing selectivity, and reliable quantification. The quantitative performance of the method was validated, and a detection limit of 0.3 ng·ml-1 for cyanidin 3-O-glucoside was determined. This newly developed method may serve to characterize and profile various anthocyanins in elderberry juices for quality control, assessment of dietary intake, and anthocyanin-based biomedical studies.