A low-cost automated titration system for colorimetric endpoint detection.

An auto titrator system was developed to accurately and precisely detect colorimetric endpoints for spectrochemical titrations. This system was constructed using inexpensive components such as a Raspberry Pi® single-board computer, 3D-printed components, and a commercially available spectral sensor. The auto titrator was evaluated by performing a standard method for determination of water hardness. Regardless of analyst experience, the auto titrator performed better than the traditional titration approach that involves manual dosing of titrant and visual detection of the endpoint. Inter-day, intra-day, inter-instrumental, and intra-instrumental validation studies were performed to establish the accuracy and precision of endpoint detection. The auto titrator eliminates the subjective bias in color perception and produces accurate and precise endpoint results.

Acute outcomes for the full US cohort of the FLASH mechanical thrombectomy registry in pulmonary embolism.

BACKGROUND: Evidence supporting interventional pulmonary embolism (PE) treatment is needed. AIMS: We aimed to evaluate the acute safety and effectiveness of mechanical thrombectomy for intermediate- and high-risk PE in a large real-world population. METHODS: FLASH is a multicentre, prospective registry enrolling up to 1,000 US and European PE patients treated with mechanical thrombectomy using the FlowTriever System. The primary safety endpoint is a major adverse event composite including device-related death and major bleeding at 48 hours, and intraprocedural adverse events. Acute mortality and 48-hour outcomes are reported. Multivariate regression analysed characteristics associated with pulmonary artery pressure and dyspnoea improvement. RESULTS: Among 800 patients in the full US cohort, 76.7% had intermediate-high risk PE, 7.9% had high-risk PE, and 32.1% had thrombolytic contraindications. Major adverse events occurred in 1.8% of patients. All-cause mortality was 0.3% at 48-hour follow-up and 0.8% at 30-day follow-up, with no device-related deaths. Immediate haemodynamic improvements included a 7.6 mmHg mean drop in mean pulmonary artery pressure (-23.0%; p<0.0001) and a 0.3 L/min/m2 mean increase in cardiac index (18.9%; p<0.0001) in patients with depressed baseline values. Most patients (62.6%) had no overnight intensive care unit stay post-procedure. At 48 hours, the echocardiographic right ventricle/left ventricle ratio decreased from 1.23±0.36 to 0.98±0.31 (p<0.0001 for paired values) and patients with severe dyspnoea decreased from 66.5% to 15.6% (p<0.0001).  Conclusions: Mechanical thrombectomy with the FlowTriever System demonstrates a favourable safety profile, improvements in haemodynamics and functional outcomes, and low 30-day mortality for intermediate- and high-risk PE.

Percutaneous mechanical thrombectomy in a real-world pulmonary embolism population: Interim results of the FLASH registry.

OBJECTIVES: The FlowTriever All-Comer Registry for Patient Safety and Hemodynamics (FLASH) is a prospective multi-center registry evaluating the safety and effectiveness of percutaneous mechanical thrombectomy for treatment of pulmonary embolism (PE) in a real-world patient population (NCT03761173). This interim analysis reports outcomes for the first 250 patients enrolled in FLASH. BACKGROUND: High- and intermediate-risk PEs are characterized by high mortality rates, frequent readmissions, and long-term sequelae. Mechanical thrombectomy is emerging as a front-line therapy for PE that enables immediate thrombus reduction while avoiding the bleeding risks inherent with thrombolytics. METHODS: The primary endpoint is a composite of major adverse events (MAE) including device-related death, major bleeding, and intraprocedural device- or procedure-related adverse events at 48 h. Secondary endpoints include on-table changes in hemodynamics and longer-term measures including dyspnea, heart rate, and cardiac function. RESULTS: Patients were predominantly intermediate-risk per ESC guidelines (6.8% high-risk, 93.2% intermediate-risk). There were three MAEs (1.2%), all of which were major bleeds that resolved without sequelae, with no device-related injuries, clinical deteriorations, or deaths at 48 h. All-cause mortality was 0.4% at 30 days, with a single death that was unrelated to PE. Significant on-table improvements in hemodynamics were noted, including an average reduction in mean pulmonary artery pressure of 7.1 mmHg (22.2%, p < 0.001). Patient symptoms and cardiac function improved through follow-up. CONCLUSIONS: These interim results provide preliminary evidence of excellent safety in a real-world PE population. Reported outcomes suggest that mechanical thrombectomy can result in immediate hemodynamic improvements, symptom reduction, and cardiac function recovery.

Feeding Sheep Cobalt and Oregano Essential Oil Alone or in Combination on Ruminal Nutrient Digestibility, Fermentation, and Fiber Digestion Combined With Scanning Electron Microscopy.

The feeding of Co lactate (Co), an essential oil blend (EO; oregano), or a combination of Co and EO (EOC) may improve nutrient digestion of corn silage-based rations. In four separate studies, Co, EO, or EOC was fed at 0, 4, and 7 g/days to nine rumen fistulated rams arranged in a replicated 3 × 3 Latin square design. The fourth study evaluated the carrier at 0, 4, and 7 g/day. In each ram, fresh ensiled corn silage, leaf, and husk were placed in individual nylon bags inserted through the ruminal cannula and removed after 48 h. Rams fed increasing carrier rates demonstrated similar (P > 0.10) nutrient digestibilities and ruminal pH and volatile fatty acid concentrations. Feeding Co at 4 and 7 g/day increased (P < 0.05) digestibility of DM (59.4, 63.9, and 62.4% for 0, 4, and 7 g/day, respectively), NDF (59.4, 63.9, and 62.4%), and hemicellulose (HC; 56.2, 63.6, and 65.9%) compared with rams fed 0 g/day, while CP digestibility (46.4, 49.9, and 57.8%) was improved (P < 0.05) in rams fed 7 g/day compared with those fed 0 and 4 g/day. Rams fed 4 g/day EO digested greater (P < 0.05) HC (64.1, 71.4, and 69.1%) than rams fed 0 g/day, while rams fed 7 g/day were intermediate and similar (P > 0.10). Rams fed the EOC combination at 4 and 7 g/day demonstrated greater (P < 0.05) digestibilities of DM (57.7, 60.0, and 60.0%), NDF (21.4, 28.8, and 27.7%), and ADF (24.3, 33.3, and 34.4%) than rams fed 0 g/day. The SEM and SM techniques visually demonstrated minor evidence of husk and leaf digestibility in rams across the three experiments when fed 0 g/day of Co, EO, or EOC; rams fed 4 g/day of Co, EO, or EOC exhibited varying visual signs of leaf digestion with some palisade tissue, spongy tissue, and whole vein structure remaining, while in rams fed 7 g/day, only the vein structure remained. Results demonstrated that feeding Co, EO, or EOC at 4 or 7 g/day enhanced ruminal nutrient digestion and fermentation parameters, which was visually confirmed via SEM and SM.

Effects of bale feeder design on hay waste, intake, and apparent diet digestibility in gestating beef cows.

Two experiments were conducted to determine the effects of feeder design on hay intake, apparent diet digestibility, and hay waste in gestating beef cows. Native tallgrass prairie hay and a protein supplement was fed throughout both experiments. In Exp. 1, 56 crossbred cows were used in a Latin square arrangement. Feeder design treatments included a conventional open bottom steel ring (OBSR), an open bottom polyethylene pipe ring (POLY); a sheeted bottom steel ring (RING), and a sheeted bottom steel ring with a basket (BASK). Cows were weighed and allotted based on BW to one of four previously grazed 2.0 ha paddocks equipped with a concrete feeding pad. Fourteen cows were assigned to each paddock and three round bales were fed consecutively within each treatment period. The cows acclimated to the feeders while the first bale was being consumed. Subsequently, hay waste data were collected while the second and third bale within each period were being consumed. Waste was measured for each bale at 24, 48, 72, and 96 h after each bale was introduced into the pen. Hay waste was significantly affected by hay feeder design with 19.7, 21.1, 12.4, and 5.5% of original bale weight wasted for OBSR, POLY, RING, and BASK, respectively (P < 0.01). There was a feeder design × day interaction (P < 0.01) with greater waste when the bale was first introduced into the pen in OBSR, POLY, and RING feeders and gradually declining thereafter, while waste from the BASK feeder was consistently low. There was a tendency (P = 0.06) for cows eating from OBSR feeders to consume less hay than cows eating from RING feeders. Feeder design did not influence apparent diet digestibility (P = 0.46). In Exp. 2, 64 crossbred cows (body weight = 590 ± 59 kg) were used to determine waste, forage intake, and apparent diet digestibility when hay was fed from a sheeted bottom steel ring (RING) or a RING feeder with a cone insert (CONE). More hay was wasted when cows were fed from RING feeders compared to CONE feeders (11.9% vs. 4.8%, P < 0.01). Feeder design had no effect on DMI or apparent digestibility (P > 0.45). Hay savings from adopting a more conservative feeder design can have a dramatic influence on hay utilization by beef cows and thus on cost of production.

Dietary Protein and Gut Microbiota Composition and Function.

Dietary protein and its metabolites, amino acids, are essential nutrients for humans and animals. Accumulated research has revealed that the gut microbiota mediate the crosstalk between protein metabolism and host immune response. Gut microbes are involved in the digestion, absorption, metabolism and transformation process of dietary protein in the gastrointestinal tract. Amino acids can be metabolized into numerous microbial metabolites, and these metabolites participate in various physiological functions related to host health and diseases. The components of dietary protein impact the gut microbiota composition and microbial metabolites. The source, concentration, and amino acid balance of dietary protein are primary factors which contribute to the composition, structure and function of gut microbes. A suitable ratio between protein and carbohydrate or even a low protein diet is recommended over a diet with protein in excess of requirements. Greater levels and undigested protein lead to an increase of pathogenic microorganism with associated higher risk of metabolic diseases. Herein, the crosstalk between dietary protein and gut microbiota composition and function is summarized, which will help to reveal the potential mechanism of gut microbes on the gastrointestinal tract health.

Maturation of Breathing-Related Inhibitory Neurotransmission in the Medulla Oblongata of the Embryonic and Perinatal Zebra Finch (Taeniopygia guttata).

The medullary portion of the embryonic zebra finch hindbrain was isolated and superfused with physiologically relevant artificial cerebral spinal fluid. This in vitro preparation produced uninterrupted rhythmic episodes of neural activity via cranial nerve IX (glossopharyngeal) from embryonic day 4 (E4) through hatching on E14. Cranial nerve IX carries motor activity to the glottis during the inspiratory phase of breathing, and we focused on the role of synaptic inhibition during the embryonic and perinatal maturation of this branchiomotor outflow. We show that spontaneous neural activity (SNA) is first observed on E4 and temporally transforms as the embryo ages. To start, SNA is dependent on the excitatory actions of GABAA and glycine. As the embryo continues to develop, GABAergic and glycinergic neurotransmission take on a modulatory role, albeit an excitatory one, through E10. After that, data show that GABAergic and glycinergic neurotransmission switches to a phenotype consistent with inhibition, coincident with the onset of functional breathing. We also report that the inhibitory action of GABAergic and glycinergic receptor gating is not necessary for the spontaneous generation of branchiomotor motor rhythms in these birds near hatching. This is the first report focusing on the development of central breathing-related inhibitory neurotransmission in birds during the entire period of embryogenesis.

Citrate Content of Bone as a Measure of Postmortem Interval: An External Validation Study.

The postmortem interval (PMI) of skeletal remains is a crucial piece of information that can help establish the time dimension in criminal cases. Unfortunately, the accurate and reliable determination of PMI from bone continues to evade forensic investigators despite concerted efforts over the past decades to develop suitable qualitative and quantitative methods. A relatively new PMI method based on the analysis of citrate content of bone was developed by Schwarcz et al. The main objective of our research was to determine whether this work could be externally validated. Thirty-one bone samples were obtained from the Forensic Anthropology Center, University of Tennessee, Knoxville, and the Onondaga County Medical Examiner's Office. Results from analyzing samples with PMI greater than 2 years suggest that the hypothetical relationship between the citrate content of bone and PMI is much weaker than reported. It was also observed that the average absolute error between the PMI value estimated using the equation proposed by Schwarcz et al. and the actual ("true") PMI of the sample was negative indicating an underestimation in PMI. These findings are identical to those reported by Kanz et al. Despite these results this method may still serve as a technique to sort ancient from more recent skeletal cases, after further, similar validation studies have been conducted.

Association of IFIH1 and pro-inflammatory mediators: Potential new clues in SLE-associated pathogenesis.

Antiviral defenses are inappropriately activated in systemic lupus erythematosus (SLE) and association between SLE and the antiviral helicase gene, IFIH1, is well established. We sought to extend the previously reported association of pathogenic soluble mediators and autoantibodies with mouse Mda5 to its human ortholog, IFIH1. To better understand the role this gene plays in human lupus, we assessed association of IFIH1 variants with soluble mediators and autoantibodies in 357 European-American SLE patients, first-degree relatives, and unrelated, unaffected healthy controls. Association between each of 135 genotyped SNPs in IFIH1 and four lupus-associated plasma mediators, IL-6, TNF-α, IFN-ß, and IP-10, were investigated via linear regression. No significant associations were found to SNPs orthologous to those identified in exon 13 of the mouse. However, outside of this region there were significant associations between IL-6 and rs76162067 (p = 0.008), as well as IP-10 and rs79711023 (p = 0.003), located in a region of IFIH1 previously shown to directly influence MDA-5 mediated IP-10 and IL-6 secretion. SLE patients and FDRs carrying the minor allele for rs79711023 demonstrated lower levels of IP-10, while only FDRs carrying the minor allele for rs76162067 demonstrated an increased level of IL-6. This would suggest that the change in IP-10 is genotypically driven, while the change in IL-6 may be reflective of SLE transition status. These data suggest that IFIH1 may contribute to SLE pathogenesis via altered inflammatory mechanisms.

Effects of variety and storage duration on the nutrient digestibility and the digestible and metabolisable energy content of maize fed to growing pigs.

The objective of this research was to determine the effects of variety and storage duration on the nutrient digestibility and the digestible (DE) and metabolisable (ME) energy content in maize when fed to growing pigs. Four maize varieties (LS1, LS2, LS3 and LS4) were hand-harvested from the same growing area in China in early October of 2012. The samples were sun dried to about 14% moisture content and then stored in the warehouse of the Fengning Pig Experiment Base at China Agricultural University for 0, 3 or 10 months. Twenty-four barrows of about 33 kg body weight were used and allotted to a completely randomised block design with four diets and six replicate pigs per diet. Pigs were individually housed in metabolic crates. The four experimental diets were formulated by mixing 96.8% of each variety of maize with 3.2% vitamins and minerals. A 5-day collection period followed a 7-day diet acclimation period. The results indicated that the DE and ME contents of maize and the apparent total tract digestibility (ATTD) of organic matter (OM), dry matter, gross energy (GE), neutral detergent fibre, acid detergent fibre (ADF), crude protein (CP) and ether extract (EE) were significantly (p < 0.05) influenced by maize variety and storage duration. With an extension of storage duration from 0 to 10 months, the DE and ME of maize and the ATTD of OM, GE, ADF, CP and EE changed in a quadratic manner (p < 0.05), and 3 months of storage exceeded 0 months of storage by 1.84%, 1.43%, 0.31%, 0.32%, 15.37%, 2.11% and 5.02%, respectively. The DE, ME of maize and the ATTD of OM, GE, ADF, CP and EE decreased by 3.67%, 6.00%, 0.97%, 1.40%, 30.54%, 3.92% and 20.93%, respectively, at 10 months of storage compared to 3 months of storage. No interaction was observed between maize variety and storage duration in DE and ME contents in maize. In conclusion, under the conditions of this study, most of the nutrient digestibility and the DE and ME contents of maize increased from 0 to 3 months and decreased from 3 to 10 months.

Dietary grape seed proanthocyanidins (GSPs) improve weaned intestinal microbiota and mucosal barrier using a piglet model.

Proanthocyanidins have been suggested as an effective antibiotic alternative, however their mechanisms are still unknown. The present study investigated the effects of grape seed proanthocyanidins on gut microbiota and mucosal barrier using a weaned piglet model in comparison with colistin. Piglets weaned at 28 day were randomly assigned to four groups treated with a control ration, or supplemented with 250 mg/kg proanthocyanidins, kitasamycin/colistin, or 250 mg/kg proanthocyanidins and half-dose antibiotics, respectively. On day 28, the gut chyme and tissue samples were collected to test intestinal microbiota and barrier function, respectively. Proanthocyanidins treated piglets had better growth performance and reduced diarrhea incidence (P < 0.05), accompanied with decreased intestinal permeability and improved mucosal morphology. Gene sequencing analysis of 16S rRNA revealed that dietary proanthocyanidins improved the microbial diversity in ileal and colonic digesta, and the most abundant OTUs belong to Firmicutes and Bacteroidetes spp.. Proanthocyanidins treatment decreased the abundance of Lactobacillaceae, and increased the abundance of Clostridiaceae in both ileal and colonic lumen, which suggests that proanthocyanidins treatment changed the bacterial composition and distribution. Administration of proanthocyanidins increased the concentration of propionic acid and butyric acid in the ileum and colon, which may activate the expression of GPR41. In addition, dietary proanthocyanidins improved the antioxidant indices in serum and intestinal mucosa, accompanied with increasing expression of barrier occludin. Our findings indicated that proanthocyanidins with half-dose colistin was equivalent to the antibiotic treatment and assisted weaned animals in resisting intestinal oxidative stress by increasing diversity and improving balance of gut microbes.

Development and validation of a simple lupus severity index using ACR criteria for classification of SLE.

OBJECTIVE: To develop a simple systemic lupus erythematosus (SLE) severity index that requires knowledge of only American College of Rheumatology (ACR) criteria and subcriteria. METHODS: This study used demographic, mortality and medical records data of 1915 patients with lupus from the Lupus Family Registry and Repository. The data were randomly split (2:1 ratio) into independent training and validation sets. A logistic regression with ridge penalty was used to model the probability of being prescribed major immunosuppressive drugs-a surrogate indicator of lupus severity. ACR criteria and subcriteria were used as predictor variables in this model, and the resulting regression coefficient estimates obtained from the training data were used as item weightings to construct the severity index. RESULTS: The resulting index was tested on the independent validation dataset and was found to have high predictive accuracy for immunosuppressive use and early mortality. The index was also found to be strongly correlated with a previously existing severity score for lupus. In addition, demographic factors known to influence lupus severity (eg, age of onset, gender and ethnicity) all showed robust associations with our severity index that were consistent with observed clinical trends. CONCLUSIONS: This new index can be easily computed using ACR criteria, which may be among the most readily available data elements from patient medical records. This tool may be useful in lupus research, especially large dataset analyses to stratify patients by disease severity, an important prognostic indicator in SLE.

Activity-dependent plasticity in the isolated embryonic avian brainstem following manipulations of rhythmic spontaneous neural activity.

When rhythmic spontaneous neural activity (rSNA) first appears in the embryonic chick brainstem and cranial nerve motor axons it is principally driven by nicotinic neurotransmission (NT). At this early age, the nicotinic acetylcholine receptor (nAChR) agonist nicotine is known to critically disrupt rSNA at low concentrations (0.1-0.5µM), which are levels that mimic the blood plasma levels of a fetus following maternal cigarette smoking. Thus, we quantified the effect of persistent exposure to exogenous nicotine on rSNA using an in vitro developmental model. We found that rSNA was eliminated by continuous bath application of exogenous nicotine, but rSNA recovered activity within 6-12h despite the persistent activation and desensitization of nAChRs. During the recovery period rSNA was critically driven by chloride-mediated membrane depolarization instead of nicotinic NT. To test whether this observed compensation was unique to the antagonism of nicotinic NT or whether the loss of spiking behavior also played a role, we eliminated rSNA by lowering overall excitatory drive with a low [K(+)]o superfusate. In this context, rSNA again recovered, although the recovery time was much quicker, and exhibited a lower frequency, higher duration, and an increase in the number of bursts per episode when compared to control embryos. Importantly, we show that the main compensatory response to lower overall excitatory drive, similar to nicotinergic block, is a result of potentiated chloride mediated membrane depolarization. These results support increasing evidence that early neural circuits sense spiking behavior to maintain primordial bioelectric rhythms. Understanding the nature of developmental plasticity in the nervous system, especially versions that preserve rhythmic behaviors following clinically meaningful environmental stimuli, both normal and pathological, will require similar studies to determine the consequences of feedback compensation at more mature chronological ages.

Studying respiratory rhythm generation in a developing bird: Hatching a new experimental model using the classic in vitro brainstem-spinal cord preparation.

It has been more than thirty years since the in vitro brainstem-spinal cord preparation was first presented as a method to study automatic breathing behaviors in the neonatal rat. This straightforward preparation has led to an incredible burst of information about the location and coordination of several spontaneously active microcircuits that form the ventrolateral respiratory network of the brainstem. Despite these advances, our knowledge of the mechanisms that regulate central breathing behaviors is still incomplete. Investigations into the nature of spontaneous breathing rhythmicity have almost exclusively focused on mammals, and there is a need for comparative experimental models to evaluate several unresolved issues from a different perspective. With this in mind, we sought to develop a new avian in vitro model with the long term goal to better understand questions associated with the ontogeny of respiratory rhythm generation, neuroplasticity, and whether multiple, independent oscillators drive the major phases of breathing. The fact that birds develop in ovo provides unparalleled access to central neuronal networks throughout the prenatal period - from embryo to hatchling - that are free from confounding interactions with mother. Previous studies using in vitro avian models have been strictly limited to the early embryonic period. Consequently, the details and even the presence of brainstem derived breathing-related rhythmogenesis in birds have never been described. In the present study, we used the altricial zebra finch (Taeniopygia guttata) and show robust spontaneous motor outflow through cranial motor nerve IX, which is first detectable on embryonic day four and continues through prenatal and early postnatal development without interruption. We also show that brainstem oscillations change dramatically over the course of prenatal development, sometimes within hours, which suggests rapid maturational modifications in growth and connectivity. We propose that this experimental preparation will be useful for a variety of studies aimed at testing the biophysical and synaptic properties of neurons that participate in the unique spatiotemporal patterns of avian breathing behaviors, especially in the context of early development.

Evaluation of processing technology for Triarrhena sacchariflora (Maxim.) Nakai for ethanol production.

The effects of dilute H2SO4 concentration, forage:sulfuric acid ratio, digestion time, and digestion temperature were evaluated to determine effects on ethanol yield of Triarrhena sacchariflora (Maxim.) Nakai. Twenty single factor experiments were conducted to evaluate H2SO4 concentration (0.5, 1.0, 1.5, 2.0, and 2.5%, w/w), forage:sulfuric acid ratio (1:6, 1:8, 1:10, 1:12, and 1:14, g/ml), digestion time (15, 30, 45, 60, and 90, min), digestion temperature (80, 100, 110, 120, and 125 °C) for 3 replicates of the 5 levels of each factor. Based on results of the single factor experiments, an incomplete factorial was designed to evaluate ethanol yield from the best combinations of single factors. Finally, the best combination was tested by enzymatic hydrolysis and fermentation experiment in selected combinations according to pretreatment results. Percentage cellulose, hemicellulose, and lignin contents of forage residue after pretreatment, and glucose and xylose concentrations of the filtrate were analyzed prior to enzymatic hydrolysis, and percentage crystallinity was observed in untreated grass and pretreated residue. In addition, the solid residues were then hydrolysed and fermented by cellulase and yeast, the concentrations of glucose and ethanol being monitored for 96 h. Results showed that the order of the effect of main effect factors was as follows: digestion temperature > dilute H2SO4 concentration > digestion time > forage:sulfuric acid ratio. The best process parameters evaluated were sulfuric acid concentration of 1.5%, forage:sulfuric acid ratio of 1:6, digestion time of 15 min, and digestion temperature of 120°C. With this combination of factors, 80% of the cellulose was hydrolysed in 96 h, and 78% converted to ethanol. The findings identified that hemicelluloses were the key deconstruction barrier for pretreatment of Triarrhena sacchariflora (Maxim.) Nakai for ethanol production. The results of this research provide evidence of appropriate combinations of processing factors for production of ethanol from Triarrhena sacchariflora (Maxim.) Nakai.

Non-destructive and selective imaging of the functionally active, pro-invasive membrane type-1 matrix metalloproteinase (MT1-MMP) enzyme in cancer cells.

Proteolytic activity of cell surface-associated MT1-matrix metalloproteinase (MMP) (MMP-14) is directly related to cell migration, invasion, and metastasis. MT1-MMP is regulated as a proteinase by activation and conversion of the latent proenzyme into the active enzyme, and also via inhibition by tissue inhibitors of MMPs (TIMPs) and self-proteolysis. MT1-MMP is also regulated as a membrane protein through its internalization and recycling. Routine immunohistochemistry, flow cytometry, reverse transcription-PCR, and immunoblotting methodologies do not allow quantitative imaging and assessment of the cell-surface levels of the active, TIMP-free MT1-MMP enzyme. Here, we developed a fluorescent reporter prototype that targets the cellular active MT1-MMP enzyme alone. The reporter (MP-3653) represents a liposome tagged with a fluorochrome and functionalized with a PEG chain spacer linked to an inhibitory hydroxamate warhead. Our studies using the MP-3653 reporter and its inactive derivative demonstrated that MP-3653 can be efficiently used not only to visualize the trafficking of MT1-MMP through the cell compartment, but also to quantify the femtomolar range amounts of the cell surface-associated active MT1-MMP enzyme in multiple cancer cell types, including breast carcinoma, fibrosarcoma, and melanoma. Thus, the levels of the naturally expressed, fully functional, active cellular MT1-MMP enzyme are roughly equal to 1 × 10(5) molecules/cell, whereas these levels are in a 1 × 10(6) range in the cells with the enforced MT1-MMP expression. We suggest that the reporter we developed will contribute to the laboratory studies of MT1-MMP and then, ultimately, to the design of novel, more efficient prognostic approaches and personalized cancer therapies.

Evaluation of sterilization methods following contamination of hamstring autograft during anterior cruciate ligament reconstruction.

PURPOSE: Inadvertent contamination of the hamstring autograft during ACL reconstruction is infrequent, but can result in significant complications. The purpose of this study is to evaluate bacterial contamination of hamstring autografts dropped onto the operating room floor and methods of graft decontamination. METHODS: Hamstring tendons were harvested from patients. Excess tendon not used in the ACL procedure was divided into 6 segments. Segments were assigned to 6 groups (A through F, N = 30 in each group): group A: uncontaminated graft immediately postharvest (control), group B: graft dropped onto the floor (5 s), group C: graft dropped onto the floor (15 s). grafts in groups D to F were dropped onto floor for 15 s then rinsed with saline (group D), bacitracin solution (group E) or chlorhexidine 4 % solution (group F) for 3 min. All grafts were sent to the microbiology laboratory for anaerobic and aerobic cultures. RESULTS: Cultures were positive in 23 % of graft segments from group A (7/30), 33 % of grafts from group B (10/30), 23 % from group C (7/30), 30 % from group D (9/30) and 3 % from both group E (1/30) and group F (1/30). Sixteen unique organisms were identified, with Staphylococcus aureus as the most common isolate. Grafts rinsed in either bacitracin solution or 4 % chlorhexidine solutions were significantly less likely to be culture positive when compared to control graft segments (p < 0.05). However, there was no significant difference between uncontaminated grafts retrieved in <5 versus 15 s from the floor. CONCLUSION: This study supports the practice of decontaminating a dropped ACL hamstring autograft using either 4 % chlorhexidine or bacitracin solution. Specimens should be retrieved sterilely and washed for at least 3 min. This study also demonstrates no advantage in retrieval time of less than 5 s as compared to 15 s for uncontaminated graft. Hamstring harvest in ACL reconstruction may result in positive cultures, thus routine soaking of the hamstring autograft in either bacitracin or 4 % chlorhexidine solution is recommended. In addition, dropped hamstring autograft can be effectively sterilized with bacitracin or 4 % chlorhexidine solution. LEVEL OF EVIDENCE: II.

Minimally invasive mitral valve replacement in a patient with previous omphalocele repair and severe scoliosis.

Mitral valve surgery in a patient with severe chest deformity and poor respiratory function can be associated with a high risk due to difficult access and postoperative respiratory failure. A 45-year-old man with scoliosis and respiratory dysfunction who had undergone previous omphalocele repairs presented with severe mitral regurgitation. Mitral valve replacement via right mini-thoracotomy was successfully performed. The minimally invasive approach was considered useful in this patient with anatomical difficulty and respiratory dysfunction.

Acceptance, panic, and partial recovery the pattern of usage of drug-eluting stents after introduction in the U.S. (a report from the American College of Cardiology/National Cardiovascular Data Registry).

OBJECTIVES: Review the use of drug-eluting stents (DES) to evaluate changes in use. BACKGROUND: The DES were approved after several small studies in carefully selected patients showed dramatic reduction in in-stent restenosis. The DES were then rapidly adopted into routine practice. In 2006, 3 years after introduction, serious concerns regarding long-term safety were raised. METHODS: We queried the American College of Cardiology/National Cardiovascular Data Registry (ACC/NCDR) CathPCI Registry. The percentage of DES used through mid-2009 was reviewed overall and in subgroups of patients categorized by lesion type, clinical factors, insurance, and hospital characteristics. Multivariable logistic models relating these covariates to DES usage were constructed for 3 relevant time intervals. RESULTS: A total of 2,247,647 coronary stent procedures were analyzed. By 2005 over 90% of first stents placed were DES. Safety concerns arising in 2006 reduced DES use to 64% of first stent placed. After publication of salutary outcomes data in 2008, usage increased to 76% by mid-2009. The logistic models demonstrated decreased likelihood of DES usage in patients with: 1) ST-segment elevation myocardial infarctions; and 2) no medical insurance. The DES usage increased for in-stent restenosis. Hospital characteristics were not associated with significant differences in DES usage. CONCLUSIONS: There was rapid adoption of DES into U.S. clinical practice. Concern for late stent thrombosis in 2006 significantly altered DES use with reductions seen in subgroups at risk for thrombosis and patients with no insurance. These rapid cyclic changes after DES introduction reinforce the need for continuous, timely reporting of outcomes data after the introduction of new technologies.

Assessment of atrial septal defects in adults comparing cardiovascular magnetic resonance with transoesophageal echocardiography.

BACKGROUND: Many adult patients with secundum-type atrial septal defects (ASDs) are able to have these defects fixed percutaneously. Traditionally, this has involved an assessment of ASD size, geometry and atrial septal margins by transoesophageal echocardiography (TOE) prior to percutaneous closure. This is a semi-invasive technique, and all of the information obtained could potentially be obtained by non-invasive cardiovascular magnetic resonance (CMR). We compared the assessment of ASDs in consecutive patients being considered for percutaneous ASD closure using CMR and TOE. METHODS: Consecutive patients with ASDs diagnosed on transthoracic echocardiography (TTE) were invited to undergo both CMR and TOE. Assessment of atrial septal margins, maximal and minimal defect dimensions was performed with both techniques. Analyses between CMR and TOE were made using simple linear regression and Bland Altman Analyses. RESULTS: Total CMR scan time was 20 minutes, and comparable to the TOE examination time. A total of 20 patients (M:F = 5:15, mean age 42.8 years +/- 15.7) were included in the analyses. There was an excellent agreement between CMR and TOE for estimation of maximum defect size (R = 0.87). The anterior inferior, anterior superior and posterior inferior margins could be assessed in all patients with CMR. The posterior superior margin could not be assessed in only one patient. Furthermore, in 1 patient in whom TOE was unable to be performed, CMR was used to successfully direct percutaneous ASD closure. CONCLUSIONS: CMR agrees with TOE assessment of ASDs in the work-up for percutaneous closure. Potentially CMR could be used instead of TOE for this purpose.