Endovascular stent-graft treatment of thoracic aortic disease.
Aneurysmal diseases of the thoracic aorta are life-threatening conditions. In such cases, stent-graft treatment has been proposed as an alternative to surgery. The morbidity and mortality associated with endovascular repair are significantly lower than those associated with open surgery. In the largest surgical series, the mortality ranged from 5% to 20%. In studies of endovascular repair, the 30-day mortality was 0%-20% and the periprocedural stroke rate was 0%-7%. Often, open surgery is prohibited in patients with these high-risk lesions; thus, in many cases endovascular treatment is the only alternative. Thoracic aortic diseases that can be treated with endovascular stent-graft placement include aneurysms, dissection, traumatic rupture, traumatic pseudoaneurysms, intramural hematoma, penetrating atherosclerotic ulcers, and aortic rupture. Thorough preprocedure imaging is essential for selecting patients, choosing the stent-graft devices, and planning the intervention. Prerequisites for endovascular stent-graft placement are an adequate neck for graft attachment and adequate vascular access. When the ascending aorta or aortic arch is involved, surgical and endovascular procedures can be combined and performed simultaneously, allowing treatment of a wider range of cases. An experienced interdisciplinary team is needed to manage such cases.


Effect of Serum Triglycerides on Clinical Outcomes in Acute Pancreatitis: Findings From a Regional Integrated Health Care System.
The aim of this study was to evaluate the effect of serum triglycerides on the development of multiple or persistent organ failure in patients with acute pancreatitis. A retrospective cohort study was conducted among patients hospitalized for acute pancreatitis between 2006 and 2013. Triglyceride levels measured before and within 72 hours of admission were compared. In addition, the effect of triglyceride levels on the development of multiple or persistent organ failure during hospitalization for acute pancreatitis was assessed. Among 2519 patients, 267 patients (10.6%) developed organ failure, of which 75 patients developed multiple system organ failure and 82 patients developed persistent organ failure. Triglyceride levels in patients who developed organ failure were initially much higher than in patients who did not develop organ failure, but by 72 hours into admission, approached levels of patients who did not develop organ failure. Approximately 8% of patients had triglyceride levels greater than 500 mg/dL, the majority of which had similarly high levels before admission. Increased triglyceride levels were associated with the development of multiple or persistent organ failure among patients hospitalized with acute pancreatitis. Patients with high triglyceride levels at the time of admission were likely to have high triglyceride levels before admission.


Lacunar syndrome associated with paradoxical brain embolism.
We report a patient who developed lacunar syndrome due to left upper pons infarction after performing leg exercises associated with paradoxical brain embolism. A 32-year-old man developed right arm weakness and moderate dysarthria following leg exercise. Brain MRI showed a paramedian pontine infarction of the left upper pons, and contrast transesophageal echocardiographic examination indicated that the patent foramen ovale was the embolic source. Simultaneous RI venography examination of the lower limbs identified deep venous thrombosis in the right leg as a paradoxical emboligenic source. We concluded that the presence of lacunar syndrome suggests that this mechanism was responsible for the paradoxical brain embolism.


The inaccuracy of total uterine volume measurements: sources of error and a proposed solution.
Inaccuracies in total intrauterine volumes calculated using the prolate ellipse equation have been reported. No previous study has examined all the sources of error. In this study, a comprehensive approach was undertaken. Measurements were obtained from scans of the pregnant uterus in the prone position using an automated water-path scanner (Octoson) and in the supine position using standard static B-mode scanners. Several conclusions could be drawn: 1) From the Octoson prone scans, uterine volumes obtained using the prolate ellipse formula were markedly different from the true uterine volumes obtained by the summation of stepped areas. This showed that the prolate ellipse formula was inaccurate. 2) From the static supine scans, many observer inconsistencies were found in uterine volumes obtained from the prolate ellipse formula. This made the prolate ellipse formula unreliable. 3) Previously published graphs calculated from the prolate ellipse equation, comparing fetal age with total intrauterine volume, were found to vary accuracy, presumably as a result of 1 and 2. A more accurate approach is proposed. Using the outer uterine wall as the boundary, the stepped area-to-volume values of transverse scans taken at 3-cm intervals were found to closely approximate true volumes, with an average error of only 3.5 per cent. Since these measurements encompass the intrauterine contents and the myometrium, it is suggested that the term "total uterine volume" be used instead of "total intrauterine volume."


Sequence analysis and in vitro maturation of five precursor 5S RNAs from Bacillus Q.
Bacillus Q, which is closely related to B. subtilis, contains at least six different precursors of 5S rRNA. The complete nucleotide sequences of four of these precursors, as well as the major part of the sequence of a fifth one, have been determined. They all contain the same 5'-terminal non-conserved segment which is to a large degree homologous with the corresponding segment of the B. subtilis p5S RNAs (Sogin, M.L., Pace, N.R., Rosenberg, M., Weissman, S.M. (1976) J. Biol. Chem. 251, 3480-3488). On the other hand the 3'-terminal non-conserved sequences of the various Bacillus Q precursors show considerable differences both in length and in nucleotide sequence, while there is also little or no homology with the 3'-terminal non-conserved sequence of the B. subtilis precursors. Bacillus Q p5S RNAs do not possess tetranucleotide repeats around the sites which are cleaved during maturation, as does B. subtilis p5S RNA. Like in B. subtilis, however, the cleavage sites are contained within a double-helical region of the precursor molecules. Crude RNAse M5 isolated from various Bacillus strains can maturate the Bacillus Q p5S RNAs with high efficiency. Despite considerable differences in primary structure between the precursors from the various strains, each RNAs M5 preparation can maturate all these precursors with about the same efficiency.


[Development of a method for the quantitative evaluation of the regional kinetics of the left ventricle].
Segmental wall motion abnormalities of the left ventricle frequently occur in ischemic heart disease. An objective, quantitative method is required to ensure the reproducibility of the assessment of left ventricular regional function, especially in evaluating the changes induced by diagnostic and therapeutic interventions. In 32 normal subjects we obtained 30 degrees right anterior oblique left ventriculograms and developed a method based on the following observations. The method should reflect the symmetric uniform motion of the left ventricular silhouette. Only actual contractile motion should be taken into account; therefore, rotatory and translational motion should be compensated for. Passive systolic movement of aortic and mitral valves accounts for the contraction of neighbouring myocardial segments. Left ventricular wall excursion is most often measured by a coordinate system: since the cavity of the left ventricle becomes relatively longer during systole, left ventricular walls contract neither toward a single central point nor toward the long axis; therefore the appropriate origin of the coordinate system will be a segment. Furthermore, as more elongated left ventricular end-diastolic silhouettes appear to show a greater extent of systolic lengthening (we show evidence of this), the length of the segment must be related to the end-diastolic shape. The basic steps of the method are: 1) identification of a symmetry line, from the aortic mid-point to the apex, by connecting the mid-point of 19 diameters perpendicular to the long axis; 2) roto-translation of the end-systolic silhouette so that the end-systolic apex and aortic mid-point lie on the symmetry line.(ABSTRACT TRUNCATED AT 250 WORDS)


Design, Synthesis, and Bioevaluation of Indole Core Containing 2-Arylidine Derivatives of Thiazolopyrimidine as Multitarget Inhibitors of Cholinesterases and Monoamine Oxidase A/B for the Treatment of Alzheimer Disease.
In continuation of our previous study to identify multitarget inhibitors of cholinesterases (ChEs) and monoamine oxidase (MAOs) isoforms, we synthesized and evaluated 2-arylidine derivatives of thiazolopyrimidine for the treatment of Alzheimer disease. Three series of compounds with different linker size and target-anchoring functional groups were synthesized. Compounds 34-37 showed excellent to good AChE and BChE inhibition potential at nanomolar to low micromolar concentration. While all the compounds showed excellent MAO-B inhibition and selectivity relative to MAO-A, compounds 25 and 36 emerged as the most potent MAO-B inhibitors of all the series of synthesized compounds with IC50 values of 0.13 μM and 0.10 μM, respectively. Furthermore, kinetic studies of inhibitor 35 showed mixed inhibition mode. Exploration of structure activity relationship (SAR) revealed the role of functionalities and length of linkers on potency. Acute toxicity evaluation showed the safety of tested compounds up to 2000 mg/kg dose. PAMPA-BBB evaluation showed BBB permeability of the tested compounds, while MTT assay performed on neuroblastoma SHSY5Y cells showed that all the tested compounds are non-neurotoxic in the tested concentrations. Docking studies showed a strong correlation with experimental in vitro results via binding orientations and interaction patterns of the synthesized compounds into the binding sites of target enzymes. We have successfully identified safe, non-neurotoxic, and blood brain barrier permeable multitarget lead compounds for the treatment of AD.


Differentiation in the protein synthesis-dependency of persistent synaptic plasticity in mossy fiber and associational/commissural CA3 synapses in vivo.
Long-term potentiation (LTP) and long-term depression (LTD) are two mechanisms involved in the long-term storage of information in hippocampal synapses. In the hippocampal CA1 region, the late phases of LTP and LTD are protein-synthesis dependent. In the dentate gyrus, late-LTP but not LTD requires protein synthesis. The protein synthesis-dependency of persistent plasticity at CA3 synapses has not yet been characterized. Here, the roles of protein transcription and translation at mossy fiber (mf) and associational/commissural (AC)- synapses were studied in freely behaving rats. In control animals, low-frequency stimulation (LFS) evoked robust LTD (>24 h), whereas high-frequency stimulation (HFS) elicited robust LTP (>24 h) at both mf-CA3 and AC-CA3 synapses. Translation inhibitors prevented early and late phases of LTP and LTD at mf-CA3 synapses. In contrast, at AC-CA3 synapses, translation inhibitors prevented intermediate/late-LTP and late-LTD only. Transcription effects were also synapse-specific: whereas transcription inhibitors inhibited late-LTP and late-LTD (>3 h) at mf-CA3 synapses, at AC-CA3 synapses, protein transcription affected early-LTP and late-LTD. These results show that the AC-CA3 and mf-CA3 synapses display different properties in terms of their protein synthesis dependency, suggesting different roles in the processing of short- and long term synaptic plasticity.


Accuracy and Efficiency of Deep-Learning-Based Automation of Dual Stain Cytology in Cervical Cancer Screening.
With the advent of primary human papillomavirus testing followed by cytology for cervical cancer screening, visual interpretation of cytology slides remains the last subjective analysis step and suffers from low sensitivity and reproducibility. We developed a cloud-based whole-slide imaging platform with a deep-learning classifier for p16/Ki-67 dual-stained (DS) slides trained on biopsy-based gold standards. We compared it with conventional Pap and manual DS in 3 epidemiological studies of cervical and anal precancers from Kaiser Permanente Northern California and the University of Oklahoma comprising 4253 patients. All statistical tests were 2-sided. In independent validation at Kaiser Permanente Northern California, artificial intelligence (AI)-based DS had lower positivity than cytology (P < .001) and manual DS (P < .001) with equal sensitivity and substantially higher specificity compared with both Pap (P < .001) and manual DS (P < .001), respectively. Compared with Pap, AI-based DS reduced referral to colposcopy by one-third (41.9% vs 60.1%, P < .001). At a higher cutoff, AI-based DS had similar performance to high-grade squamous intraepithelial lesions cytology, indicating a risk high enough to allow for immediate treatment. The classifier was robust, showing comparable performance in 2 cytology systems and in anal cytology. Automated DS evaluation removes the remaining subjective component from cervical cancer screening and delivers consistent quality for providers and patients. Moving from Pap to automated DS substantially reduces the number of colposcopies and also achieves excellent performance in a simulated fully vaccinated population. Through cloud-based implementation, this approach is globally accessible. Our results demonstrate that AI not only provides automation and objectivity but also delivers a substantial benefit for women by reduction of unnecessary colposcopies.


Commentary: Can Automated Blood Culture Systems Be Both New and Improved?
Automated continuous monitoring blood culture (CMBC) systems are a cornerstone of the clinical microbiology laboratory. Despite the critical role of these systems in diagnosing life-threatening bloodstream infections, their core technologies and performance characteristics have remained largely unchanged since their introduction in the 1990s. This stability and uniformity have enabled the development of quality benchmarks, such as percent positivity and contamination rate; downstream diagnostics, such as direct identification and susceptibility testing of microorganisms in positive cultures; and clinical guidelines based on time to positivity or duration of bacteriemia. In this issue of the Journal of Clinical Microbiology, Chavez et al. (J Clin Microbiol 60:e02261-21. 2021, https://doi.org/10.1128/JCM.02261-21) built on a prior study to examine clinical impacts following the introduction of a new blood culture system which boasts enhanced organism recovery and more rapid time to detection of positive blood cultures. While one might assume that these "improvements" would result in clinical benefits, the authors uncovered some unexpected consequences associated with altering long-accepted performance characteristics. Their central finding was that implementation of the new CMBC system did result in alterations to the management of patients with S. aureus bacteremia; however, this did not have any overall consequences for patient outcomes.


An ab initio multireference perturbation theory study on the manganese dimer.
The potential energy curves of the ground state and of some excited states of the manganese dimer have been calculated over a wide range of internuclear distances using the second order n-electron valence state perturbation theory applied to a complete active space self-consistent field reference wave function. The ground state of Mn(2), for which also the third order NEVPT has been used, is calculated to be a singlet belonging to the Sigma(g) (+) symmetry, characterized by a large equilibrium internuclear distance R(e) of 3.7-3.8 A, by a low dissociation energy D(e) of 0.07-0.08 eV, and by a small harmonic frequency omega(e) of 43 cm(-1). The experimental evidence that Mn(2) is a van der Waals molecule is thus confirmed. Among the excited states, (11)Pi(u), which is usually indicated as the ground state by density functional theory studies, appears as a low-lying state with R(e)=2.50 A, D(e)=1.35 eV, and omega(e)=246 cm(-1).


IL-17B: A new area of study in the IL-17 family.
The interleukin (IL)-17 superfamily, a relatively new family of cytokines, consists of six ligands (from IL-17A to IL-17F), which bind to five receptor subtypes (from IL-17RA to IL-17RE) and induce downstream signaling. IL-17A, a prototype member of this family, has been reported to be involved in the pathogenesis of allergies, autoimmune diseases, allograft transplantations, and malignancies. Unlike IL-17A, which is mainly produced by T helper 17 cells, IL-17B is widely expressed in various tissues. Recently, the biological function of IL-17B in diseases, particularly tumors, has attracted the attention of researchers. We previously reported that the expression of IL-17RB increased in gastric cancer tissues and demonstrated that IL-17B/IL-17RB signaling plays a critical role in gastric tumor progression. However, studies on IL-17B are scant. In this review, we detail the structural characteristics, expression patterns, and biological activities of IL-17B and its potential role in the pathogenesis of diseases.


Detection of Merkel Cell Polyomavirus in Seborrheic Keratosis.
Seborrheic keratosis (SK) is the most common benign cutaneous neoplasm. A subset shows increased p16 expression. Since SK shares several features with verruca vulgaris, e.g., increased p16 expression, human papillomaviruses (HPV) have been suggested as possible causal agents. However, a relevant association could not be established between HPV and SK. In the present study we aimed to investigate the presence of Merkel cell polyomavirus (MCPyV) in relation to p16 expression in SK. P16 expression was investigated using immunohistochemistry (IHC). Presence of MCPyV was assessed in 23 formalin-fixed paraffin-embedded tissue samples of SK by molecular techniques (i.e., PCR and FISH) and IHC. 16/23 SK showed strong to moderate p16 expression. 6/23 of SK were MCPyV positive by PCR which was confirmed by FISH. Of interest, two samples with strong FISH signals also showed MCPyV expression as tested by IHC. Samples with weaker signal intensity were negative in IHC. P16 expression was not associated with the presence of MCPyV. Concluding, the detection of MCPyV DNA by PCR and FISH in SK reflects the widespread prevalence of MCPyV in the skin. However, low detection rates exclude MCPyV as a major pathogenic factor in SK, most likely representing a coincidental infection. P16 IHC does not appear as useful adjunctive surrogate marker for the presence of MCPyV in SK.


Upper airway response to electrical stimulation of the genioglossus in obstructive sleep apnea.
Contraction of the genioglossus (GG) has been shown to improve upper airway patency. In the present study, we evaluated responses in upper airway pressure-flow relationships during sleep to electrical stimulation (ES) of the GG in patients with obstructive sleep apnea. Five patients with chronically implanted hypoglossal nerve (HG) electrodes and nine patients with fine-wire electrodes inserted into the GG were studied. Airflow was measured at multiple levels of nasal pressure, and upper airway collapsibility was defined by the nasal pressure below which airflow ceased ["critical" pressure (Pcrit)]. ES shifted the pressure-flow relationships toward higher flow levels in all patients over the entire range of nasal pressure applied. Pcrit decreased similarly during both HG-ES and GG-ES (deltaPcrit was 3.98 +/- 2.31 and 3.18 +/- 1.70 cmH2O, respectively) without a significant change in upstream resistance. The site of collapse (velo- vs. oropharynx) did not influence the response to GG-ES. Moreover, ES-induced reductions in the apnea-hypopnea index of the HG-ES patients were associated with substantial decreases in Pcrit. Our findings imply that responses in apnea severity to HG-ES can be predicted by characterizing the patient's baseline pressure-flow relationships and response to GG-ES.


A harsh environment wireless pressure sensing solution utilizing high temperature electronics.
Pressure measurement under harsh environments, especially at high temperatures, is of great interest to many industries. The applicability of current pressure sensing technologies in extreme environments is limited by the embedded electronics which cannot survive beyond 300 °C ambient temperature as of today. In this paper, a pressure signal processing and wireless transmission module based on the cutting-edge Silicon Carbide (SiC) devices is designed and developed, for a commercial piezoresistive MEMS pressure sensor from Kulite Semiconductor Products, Inc. Equipped with this advanced high-temperature SiC electronics, not only the sensor head, but the entire pressure sensor suite is capable of operating at 450 °C. The addition of wireless functionality also makes the pressure sensor more flexible in harsh environments by eliminating the costly and fragile cable connections. The proposed approach was verified through prototype fabrication and high temperature bench testing from room temperature up to 450 °C. This novel high-temperature pressure sensing technology can be applied in real-time health monitoring of many systems involving harsh environments, such as military and commercial turbine engines.


Transmission spectra of sausage-like microresonators.
We experimentally develop a sausage-like microresonator (SLM) by making two microtapers on a single-mode fiber, and study whispering-gallery modes (WGMs) in SLMs with different lengths. The transmission spectra from 1530 nm to 1550 nm of several SLMs are presented and SLMs with different lengths are shown to have different transmission features. The maximal Q factor observed in the SLMs is 3.8 * 10(7). For comparison, the transmission spectrum of a fiber cylinder microresonator is given and the maximal Q factor achieved in the fiber microcylinder resonator is 1.7 * 10(7). The strain tuning of the SLM is also demonstrated.


Roles of nuclear NPY and NPY receptors in the regulation of the endocardial endothelium and heart function.
It is now well accepted that the heart is a multifunctional organ in which endothelial cells, and more particularly endocardial endothelial cells (EECs), seem to play an important role in regulating and maintaining cardiac excitation-contraction coupling. Even if major differences exist between vascular endothelial cells (VECs) and EECs, all endothelial cells including EECs release a variety of auto- and paracrine factors such as nitric oxide, endothelin-1, angiotensin II, and neuropeptide Y. All these factors were reported to affect cardiomyocyte contractile performance and rhythmicity. In this review, findings on the morphology of EECs, differences between EECs and other types of endothelial cells, interactions between EECs and the adjacent cardiomyocytes, and effects of NPY on the heart will be presented. We will also show evidence on the presence and localization of NPY and the Y1 receptor in the endocardial endothelium and discuss their role in the regulation of cytosolic and nuclear free calcium.


Hybrid multi-chip assembly of optical communication engines by in situ 3D nano-lithography.
Three-dimensional (3D) nano-printing of freeform optical waveguides, also referred to as photonic wire bonding, allows for efficient coupling between photonic chips and can greatly simplify optical system assembly. As a key advantage, the shape and the trajectory of photonic wire bonds can be adapted to the mode-field profiles and the positions of the chips, thereby offering an attractive alternative to conventional optical assembly techniques that rely on technically complex and costly high-precision alignment. However, while the fundamental advantages of the photonic wire bonding concept have been shown in proof-of-concept experiments, it has so far been unclear whether the technique can also be leveraged for practically relevant use cases with stringent reproducibility and reliability requirements. In this paper, we demonstrate optical communication engines that rely on photonic wire bonding for connecting arrays of silicon photonic modulators to InP lasers and single-mode fibres. In a first experiment, we show an eight-channel transmitter offering an aggregate line rate of 448 Gbit/s by low-complexity intensity modulation. A second experiment is dedicated to a four-channel coherent transmitter, operating at a net data rate of 732.7 Gbit/s - a record for coherent silicon photonic transmitters with co-packaged lasers. Using dedicated test chips, we further demonstrate automated mass production of photonic wire bonds with insertion losses of (0.7 ± 0.15) dB, and we show their resilience in environmental-stability tests and at high optical power. These results might form the basis for simplified assembly of advanced photonic multi-chip systems that combine the distinct advantages of different integration platforms.


Demographic, Chemical, and Helicobacter pylori Positivity Assessment in Different Types of Gallstones and the Bile in a Random Sample of Cholecystectomied Iranian Patients with Cholelithiasis.
The occurrence of stones in the gallbladder or common bile duct and the symptoms and complications they cause is called gallstone disease. The symptoms of gallstone disease range from mild, nonspecific symptoms to a severe right quadrant abdominal pain. Characteristics of gallstone types in an Iranian population have not been well studied before and there are very limited studies on the demographic pattern of stone types in our country, so this study is one of the first studies on its kind on the epidemiology of gallstone types in Iran. As information on chemical components of the stone will help in the management and prevention of gallstones, in this study, we aimed to do chemical component analysis of gallstones including cholesterol, bilirubin, and calcium. Given the conflicting reports about the relationship between H. pylori infections and gallstone formation, this study aimed to investigate the relationship between H. pylori positivity in the bile specimen of Iranian patients with cholelithiasis and formation and type of stone. This prospective study reviewed a total of 196 patients who underwent cholecystectomy for symptomatic cholelithiasis at Shahid Beheshti Training and Research Hospital affiliated to the Yasuj University of Medical Sciences between September 2015 and May 2018. Chemical analysis of gallstone components performed using the colorimetry method. Microbiological analysis for H. pylori was done using the OnSite H. pylori Ag Rapid Test on the bile sample. For the validation test of bile, the H. pylori Rapid Stool Ag Test on stool was used, and Cohen's Kappa statistical analysis was done next. There were significant associations between the stone types and age, chemical composition of the stones such as calcium, cholesterol, and bilirubin levels, and also H. pylori positivity and cholesterol and bilirubin levels; however, no significant association was found between the stone types and sex, H. pylori positivity and age, sex, stone types, and calcium level. The main bile and validity tests were matched to the substantial agreement according to Cohen's Kappa analysis. The most common drugs used were proton pump inhibitors, nonsteroidal anti-inflammatory drugs, antihypertensive drugs, and oral contraceptives. This study suggested that the chemical composition of the stones could predict the presence of bacteria, there is no correlation between H. pylori and gallstone formation, and some of the drugs could be predisposing factors for gallstones. This work provides an objective basis for further research into gallbladder stone formation; meanwhile, it has great significance in the treatment and prevention of gallbladder stones. Trial registration. The project was found to be in accordance to the ethical principles and the national norms and standards for conducting research in Iran with the approval ID IR.YUMS.REC.1399.147 and date 2020.09.23, and this project is the result of a residency dissertation to obtain the specialty in general surgery, which has been registered with the research project number 960159 in the Vice Chancellor for Research and Technology Development of the Yasuj University of Medical Sciences, Yasuj, Iran, URL: https://ethics.research.ac.ir/EthicsProposalViewEn.php?id=160634.


Rapid detection and identification of bacterial pathogens by using an ATP bioluminescence immunoassay.
Rapid identification of viable bacterial contaminants in food products is important because of their potential to cause disease. This study examined a method for microbial detection by using a combined ATP bioluminescence immunoassay. Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium were selected as target organisms because of their implication in foodborne illness. Various matrices containing the target cells were examined, including ground beef homogenate, apple juice, milk, and phosphate-buffered saline. Specific antibodies were immobilized on the surface of 96-well plates, and then the sample matrices containing target cells in the wells were incubated. Sample matrix (no cells) was used to establish background. The plates were washed, and the wells were incubated with BacTiter-Glo reagent in Mueller-Hinton II broth. Bioluminescent output was measured with the GloMax 96 luminometer. Signal-to-noise ratios were calculated, resulting in a limit of detection of 10(4) CFU/ml for both E. coli O157:H7 and Salmonella Typhimurium. The limit of detection for both species was not affected by the presence of nontarget cells. The various sample matrices did not affect signal-to-noise ratios when E. coli O157:H7 was the target. A weak matrix effect was observed when Salmonella Typhimurium was the target. A strong linear correlation was observed between the number of cells and luminescent output over 4 orders of magnitude for both species. This method provides a means of simultaneously detecting and identifying viable pathogens in complex matrices, and could have wider application in food microbiology.


[Nuclear medicine diagnostic localization in primary aldosteronism and Cushing syndrome].
The main indication for 131J-cholesterol scintigraphy is primary aldosteronism. The preoperative localization of aldosterone - producing adenomas may be obtained with the non-invasive NP-59 - scintigraphy in 75-85% of the patients. Idiopathic bilateral adrenal hyperplasia is diagnosed correctly in 90-100% of the cases. In Cushing's syndrome scintigraphy of the adrenal cortex is indicated only in specific cases such as very small adenomas, primary nodular adrenal hyperplasia and/or for the localization of ectopic adrenal cortex tissue. A false diagnosis may be caused by asymmetric uptake of the radioisotope in idiopathic bilateral hyperplasia or by unspecific absorption in non - adenomatous adrenal cortex tissue with symmetric uptake in patients with unilateral adenomas.


A three-dimensional ex vivo tri-culture model mimics cell-cell interactions between acute myeloid leukemia and the vascular niche.
Ex vivo studies of human disease, such as acute myeloid leukemia, are generally limited to the analysis of two-dimensional cultures which often misinterpret the effectiveness of chemotherapeutics and other treatments. Here we show that matrix metalloproteinase-sensitive hydrogels prepared from poly(ethylene glycol) and heparin functionalized with adhesion ligands and pro-angiogenic factors can be instrumental to produce robust three-dimensional culture models, allowing for the analysis of acute myeloid leukemia development and response to treatment. We evaluated the growth of four leukemia cell lines, KG1a, MOLM13, MV4-11 and OCI-AML3, as well as samples from patients with acute myeloid leukemia. Furthermore, endothelial cells and mesenchymal stromal cells were co-seeded to mimic the vascular niche for acute myeloid leukemia cells. Greater drug resistance to daunorubicin and cytarabine was demonstrated in three-dimensional cultures and in vascular co-cultures when compared with two-dimensional suspension cultures, opening the way for drug combination studies. Application of the C-X-C chemokine receptor type 4 (CXCR4) inhibitor, AMD3100, induced mobilization of the acute myeloid leukemia cells from the vascular networks. These findings indicate that the three-dimensional tri-culture model provides a specialized platform for the investigation of cell-cell interactions, addressing a key challenge of current testing models. This ex vivo system allows for personalized analysis of the responses of patients' cells, providing new insights into the development of acute myeloid leukemia and therapies for this disease.


Epizootic disseminated toxoplasmosis in captive slender-tailed meerkats (Suricata suricatta).
Fatal disseminated toxoplasmosis was diagnosed in seven captive slender-tailed meerkats (Suricata suricatta) according to clinicopathologic findings and immunohistochemistry. Five of nine meerkats died during an outbreak in late 1994. These included four kits (2.5 to 4.5 months old) and a 4-year-old meerkat. Two other meerkats, both adults, died in 1992 and 1995. Respiratory insufficiency (4/7) and incoordination (3/7) were the most consistent clinical signs. although two of seven meerkats died unexpectedly. At necropsy, the lungs were reddened and noncollapsed (6/7), and had multiple pale round foci (4/7). Yellow foci of necrosis in mesenteric lymph nodes (4/7), splenomegaly (3/7), and hydropericardium (3/7) were other common gross findings. Microscopically, interstitial pneumonia was present in all seven meerkats, being acute to subacute in six of them. Type 2 pneumocyte hyperplasia, aggregates of foamy macrophages, and giant cells were consistently seen. Multifocal to locally extensive necrosis of mesenteric lymph nodes (4/7), mild to severe multifocal necrotizing hepatitis (5/6), and mild nonsuppurative encephalitis (4/6) were also seen. Toxoplasma-like organisms were consistently associated with these lesions and were stained by the avidin biotin peroxidase procedure with an antiserum that does not cross-react with Neospora caninum. Meerkats were most likely infected after an oral, primary exposure to Toxoplasma. Several observations indicate that meerkats may be highly susceptible to toxoplasmosis.


Heteroscorpionate ligands based on bis(pyrazol-1-yl)methane: design and coordination chemistry.
Scorpionates represent one of the most versatile types of tridentate ligand that can coordinate to a wide variety of elements, e.g. from early to late transition metals, and the coordination chemistry of these systems has developed greatly in recent years. This Perspective gives an account of studies on the following aspects: (1) the preparative methods for a new class of heteroscorpionate [RR'C(pz)2] ligand derived from bis(pyrazol-1-yl)methane and (2) the description of metal complexes containing these ligands, examples of which incorporate a range of different metals from the Periodic Table.


Case report: adult-onset manifesting heterozygous glycogen storage disease type IV with dilated cardiomyopathy and absent late gadolinium enhancement on cardiac magnetic resonance imaging.
Glycogen storage disease type IV (GSD IV; Andersen's disease) is a rare autosomal recessive disease caused by mutation in the GBE1 gene. Presentation of GSD IV varies on a continuum of severity and symptomatology ranging from neonatal death to mild adult-onset disease with variable involvement of hepatic, muscular, neurologic, dermatologic, and cardiac systems. Cardiomyopathy seen in GSD IV is also heterogeneous and its appearance on cardiac magnetic resonance imaging (CMR) is rarely described. A 29-year-old man without previous medical history was admitted to our facility multiple times over 2 years for focal sensorimotor deficits, gout arthropathy, chronic hyperlactataemia and hyperuricaemia, and severe decompensated non-ischaemic cardiomyopathy complicated by episodes of thromboembolic organ infarction. Echocardiography and CMR showed severe biventricular failure with the presence of intraventricular thrombi with increased right ventricular trabeculation and absent late gadolinium enhancement. He underwent muscle biopsy which showed prominent glycogen in skeletal muscle followed by genetic testing showing a single heterozygous splicing mutation c.993-1G>T found at the junction of intron 7 and exon 8 of the GBE1 gene which had not previously been reported and was predicted to be pathologic. He was referred to a tertiary care centre with glycogen storage disease specialists but expired prior to establishing care at that facility. Discovery of GSD IV in our patient was unexpected due to a highly variant clinical presentation. Our case stresses the clinical heterogeneity of GSD IV and the importance of genetic sequencing studies in the evaluation of potential glycogen storage disease.


Synchronization to metrical levels in music depends on low-frequency spectral components and tempo.
Previous studies have found relationships between music-induced movement and musical characteristics on more general levels, such as tempo or pulse clarity. This study focused on synchronization abilities to music of finely-varying tempi and varying degrees of low-frequency spectral change/flux. Excerpts from six classic Motown/R&B songs at three different tempos (105, 115, and 130 BPM) were used as stimuli in this experiment. Each was then time-stretched by a factor of 5% with regard to the original tempo, yielding a total of 12 stimuli that were presented to 30 participants. Participants were asked to move along with the stimuli while being recorded with an optical motion capture system. Synchronization analysis was performed relative to the beat and the bar level of the music and four body parts. Results suggest that participants synchronized different body parts to specific metrical levels; in particular, vertical movements of hip and feet were synchronized to the beat level when the music contained large amounts of low-frequency spectral flux and had a slower tempo, while synchronization of head and hands was more tightly coupled to the weak flux stimuli at the bar level. Synchronization was generally more tightly coupled to the slower versions of the same stimuli, while synchronization showed an inverted u-shape effect at the bar level as tempo increased. These results indicate complex relationships between musical characteristics, in particular regarding metrical and temporal structure, and our ability to synchronize and entrain to such musical stimuli.


Exercise training increases GAD65 expression, restores the depressed GABAA receptor function within the PVN and reduces sympathetic modulation in hypertension.
GABAergic inhibitory input within the paraventricular hypothalamic nucleus (PVN) plays a key role in restraining sympathetic outflow. Although experimental evidence has shown depressed GABAA receptor function plus sympathoexcitation in hypertension and augmented GABA levels with reduced sympathetic activity after exercise training (T), the mechanisms underlying T-induced effects remain unclear. Here we investigated in T and sedentary (S) SHR and WKY: (1) time-course changes of hemodynamic parameters and PVN glutamic acid decarboxylase (GAD) isoforms' expression; (2) arterial pressure (AP) and heart rate (HR) responses, sympathetic/parasympathetic modulation of heart and vessels and baroreflex sensitivity to GABAA receptor blockade within the PVN. SHR-S versus WKY-S exhibited higher AP and HR, increased sympathetic reduced parasympathetic modulation, smaller baroreflex sensitivity, and reduced PVN GAD65 immunoreactivity. SHR-T and WKY-T showed prompt maintained increase (2-8 weeks) in GAD65 expression (responsible for GABA vesicular pool synthesis), which occurred simultaneously with HR reduction in SHR-T and preceded MAP fall in SHR-T and resting bradycardia in WKY-T. There was no change in GAD67 expression (mainly involved with GABA metabolic pool). Resting HR in both groups and basal MAP in SHR were negatively correlated with PVN GAD65 expression. Normalized baroreflex sensitivity and autonomic control observed only in SHR-T were due to recovery of GABAA receptor function into the PVN since bicuculline administration abolished these effects. Data indicated that training augments in both groups the expression/activity of GABAergic neurotransmission within presympathetic PVN neurons and restores GABAA receptors' function specifically in the SHR, therefore strengthening GABAergic modulation of sympathetic outflow in hypertension.


Dimethyl sulfide removal from synthetic waste gas using a flat poly(dimethylsiloxane)-coated composite membrane bioreactor.
The reduction of volatile organic sulfur emissions should be completely as they cause odor nuisance, even when they are emitted in very small amounts. In general, biofilters are applied for odor reduction, but their operational control is limited. A new biotechnique for the treatment of complex emissions is the use of membranes integrated within bioreactors (MBRs). In this study, the reactor performance of MBRs for removal of dimethyl sulfide (DMS) as a model compound is presented. Composite membranes with a poly(dimethylsiloxane) (PDMS)-coating layer were used. The MBRs were inoculated with the sulfur-degrading culture Hyphomicrobium VS or a suspension of Hyphomicrobium VS, ammonium-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB) to colonize the PDMS-coating layer. Although inoculation with AOB and NOB might give rise to competition for space on the membrane, their presence in the MBRs appeared to be positive as they co-oxidize DMS. Dimethyl sulfide elimination depended on the inoculum type, DMS inlet concentration, gas residence time, and membrane polymer. For equal loading rates, the elimination capacity (EC) increased at larger gas residence times and inlet concentrations. The maximum EC obtained with the MBRs was 4.8 kg of DMS x m(-3) x d(-1). This value is higher than any reported figure for biofilters and biotrickling filters.


Red blood cell rheological alterations in a rat model of ischemia-reperfusion injury.
Red blood cell (RBC) deformability and aggregation characteristics were investigated in an experimental model of ischemia-reperfusion injury. Ischemia was produced in rat hind limb by occluding the femoral artery for 10 minutes, followed by reperfusion. Blood samples were obtained either following the ischemia or 15 minutes after reperfusion. RBC deformability measured by ektacytometry was found to be significantly impaired immediately after the end of ischemic period in the blood samples obtained from femoral vein of the ischemic limb, while there was no significant difference after 15 minutes of reperfusion. In contrast, RBC aggregability was found to be decreased only after the reperfusion period and this alteration was not only limited to the blood returning from the ischemic limb but was also observed in the samples obtained from non-ischemic, contralateral hind limb, indicating a systemic alteration. RBC electrophoresis studies suggested that the altered aggregability might be related to altered RBC surface properties including increased RBC surface charge density.


Major Depression Associated With Widowhood.
The authors look at the frequency of depressive symptoms and syndromes associated with spousal bereavement in late life and assess both the course and associated morbidity of those syndromes over a 2-year period. The prevalence of major depressive syndromes ranges from 24% 2 months after the spouse's death to 14% at 25 months. Major depression was associated with poor role functioning, interpersonal difficulties, poor medical health, and increased medical treatment. Risk factors for continuing major depression 25 months after the death of a spouse include, in descending order of importance, early post-bereavement depressive syndromes, intensity of early depressive symptoms, family history of depression, alcohol consumption, medical health, and suddenness of the death.


[Longitudinal evaluation of aerobic capacity determined by torque auto-controlled system with the feedback of photoelectric pulse].
We have previously developed a unique and simple procedure for assessing cardiorespiratory fitness. The present investigation was conducted to evaluate whether an aerobic index determined by torque auto-controlled system with the feedback of photoelectric pulse could sufficiently approximate the cardiorespiratory fitness represented by anaerobic threshold (AT) and maximal oxygen uptake (VO2max). Analysis of the cross-sectional data indicated that the aerobic score (AS: K (WR/Wt)/HR; where WR = mean work rate during 12-min cycling, Wt = weight, and HR = mean heart rate during 12-min cycling) determined by the torque auto-controlled system was significantly correlated with directly measured VO2/AT (r = 0.922, 76 males; r = 0.814, 34 females). Cross-validity of the predicted VO2max from the AS was significant and sufficiently high (r = 0.949, 31 males) for use in the general public. In addition, the effects of cycling or jogging training on the AS were longitudinally investigated on 17 females and 1 male. Major effects of the training were significant improvements in the AS, VO2max, and VO2/AT. Delta percentage change [(post-value - pre-value)/pre-value; delta %] in the AS was closely associated (r = 0.718, 8 females) with delta % in VO2/AT. It appears likely from the present investigation that information obtained through the use of our unique system (i.e., the AS) could provide considerably reliable estimate of cardiorespiratory fitness in both males and females.


"Some guys have all the luck": mate preference influences paced-mating behavior in female rats.
In the present study, mating behavior was observed in female rats that were given the opportunity to mate with two males simultaneously. Three groups of females were tested: 1) sexually naïve, naturally cycling rats in proestrous; 2) sexually naïve, hormone-primed, ovariectomized (OVX) rats; and 3) hormone-primed, OVX rats tested 1 week after sexual receptivity testing. One male rat was determined to be the preferred male for each female, if she spent more time with him during a mating test. Independent of sexual experience, female rats were less likely to leave their preferred male than their non-preferred male following intromissions. However, when they left their preferred male, they returned to him faster than to their non-preferred male. This effect of preference was slightly more robust in the OVX rats. When female rats from Group 2 were tested with the same pair of males for 3 additional tests, each female's preference for a particular male was stable. That is, a female rat preferred the same male in approximately 3 out of the 4 tests, which is more likely than would be expected by chance. In a final experiment, pairs of male rats were tested with different females once weekly to determine if different females would prefer particular males consistently. Although no male rat was preferred by all females, females consistently preferred the same male from each pair during approximately 70% of the tests. In conclusion, female mate preference may have adaptive significance for the reproductive success of rats.


Management of patients with asymptomatic moderate aortic stenosis undergoing coronary artery bypass grafting.
Manual decalcification of the aortic valve was performed systematically in a prospective series of patients with asymptomatic moderate aortic stenosis (AS) undergoing coronary artery bypass grafting (CABG). This study addressed two main issues: (i) whether aortic valve decalcification is a good option to relieve moderate AS; and (ii) whether the natural progression of AS may be delayed by manual valve debridement when surgery is indicated for coronary disease. Between October 1997 and March 2001, 14 adult patients with moderate AS underwent concomitant surgical repair of the aortic valve during CABG. Manual valve debridement with restoration of cusp mobility was attempted. Calcified deposits were removed by careful dissection. All patients underwent myocardial revascularization; a mean of 2.38 grafts was performed per patient. Immediately after surgery, mean aortic valve area index (AVAI) was improved, from 0.56+/-0.12 to 1.43+/-0.25 cm2/m2. Patients with the slowest recalcification rates were those with a postoperative/preoperative AVAI ratio of 1.6 to 2.4, those in whom the degree of postoperative aortic insufficiency was very similar to the degree of preoperative regurgitation, and those in whom the preoperative AVAI was >0.55 cm2/m2. Manual aortic valve debridement for moderate AS is a good option when surgery must be performed for coronary disease; the best results were obtained in patients with senile stenosis of a tricuspid aortic valve with an AVAI of 0.55-0.9 cm2/m2.


Increased leucocyte Na-K ATPase in obesity: reversal following weight loss.
Ouabain-sensitive 86Rb influx and [3H] ouabain binding capacity were investigated in the leucocytes of 17 obese patients and 15 control subjects. Both were significantly increased in the obese when compared with controls. Following dietary restriction and a 4% to 5% weight reduction in the obese over 2 weeks, [3H] ouabain binding and ouabain-sensitive 86Rb influx (a model for K+ influx) decreased to levels similar to those in controls. This shows that the number of Na-K ATPase sites on leucocyte membranes of the obese are significantly increased and that this is associated with accelerated 86Rb transport. Since both of these indices decreased following 4% to 5% reduction in body weight while the patients were still obese, increased Na-K ATPase is neither a marker of nor cardinal to the pathogenesis of obesity. We conclude that (1) increase in Na-K ATPase units and 86Rb influx are not characteristic of obesity itself and (2) dietary restriction over the short-term with limited weight reduction restores Na-K ATPase units and 86Rb influx to normal.


Galectin-3 in patients with heart failure with preserved ejection fraction: results from the Aldo-DHF trial.
Galectin-3 is a marker of myocardial fibrosis and mediates aldosterone-induced cardiovascular inflammation and fibrosis. Characteristics of galectin-3 and its response to spironolactone have not been evaluated in heart failure with preserved ejection fraction (HFpEF). The aim of this study was to determine the association between galectin-3 levels and patient characteristics in HFpEF; to evaluate the interaction between spironolactone and galectin-3 levels; and to assess the association between galectin-3 and clinical outcomes. Aldo-DHF investigated spironolactone 25 mg once daily vs. placebo for 12 months in patients with NYHA class II-III, LVEF ≥50%, grade ≥ I diastolic dysfunction, and peakVO2 ≤ 25 mL/kg/min. Galectin-3 levels were obtained at baseline, and at 6 and 12 months. The association between baseline galectin-3, change in galectin-3, and all-cause death or hospitalization was evaluated, and the interaction between galectin-3 and treatment was assessed. Median baseline galectin-3 was 12.1 ng/mL. After multivariable adjustment, baseline galectin-3 inversely correlated with peak VO2 (P = 0.021), 6 min walk distance (P = 0.002), and Short Form 36 (SF-36) physical functioning (P = 0.001), and directly correlated with NYHA class (P = 0.007). Baseline NT-proBNP correlated with E/e' velocity ratio (P ≤ 0.001), left atrial volume index (P < 0.001), and LV mass index (P = 0.009). Increasing galectin-3 at 6 or 12 months was associated with all-cause death or hospitalization independent of treatment arm [hazard ratio (HR) 3.319, 95% confidence interval (CI) 1.214-9.07, P = 0.019] and NT-proBNP (HR 3.127, 95% CI 1.144-8.549, P = 0.026). Spironolactone did not influence galectin-3 levels. Galectin-3 levels are modestly elevated in patients with stable HFpEF and relate to functional performance and quality of life. Increasing galectin-3 was associated with worse outcome, independent of treatment or NT-proBNP.


Shedding of APP limits its synaptogenic activity and cell adhesion properties.
The amyloid precursor protein (APP) plays a central role in Alzheimer's disease (AD) and has essential synapse promoting functions. Synaptogenic activity as well as cell adhesion properties of APP presumably depend on trans-cellular dimerization via its extracellular domain. Since neuronal APP is extensively processed by secretases, it raises the question if APP shedding affects its cell adhesion and synaptogenic properties. We show that inhibition of APP shedding using cleavage deficient forms of APP or a dominant negative α-secretase strongly enhanced its cell adhesion and synaptogenic activity suggesting that synapse promoting function of APP is tightly regulated by α-secretase mediated processing, similar to other trans-cellular synaptic adhesion molecules.


Short-term impact of a neighbourhood-based intervention on mental health and self-rated health in Hamilton, Ontario, Canada.
The Hamilton Neighbourhoods Study aimed to measure the short-term impact of a neighbourhood-based intervention known as the City of Hamilton's Neighbourhood Action Strategy on health and neighbourhood outcomes. A quasi-experimental study with 881 intervention participants across six targeted neighbourhoods and 173 control participants was conducted to investigate changes in self-rated health and mental health from baseline to follow-up. There was evidence of small improvements in mental health in two neighbourhoods, but there was no change in self-rated health. Place-based interventions aimed at high poverty neighbourhoods may have only modest impacts on health in the short-term.


Structural determinants at the M2 muscarinic receptor modulate the RGS4-GIRK response to pilocarpine by impairment of the receptor voltage sensitivity.
Membrane potential controls the response of the M2 muscarinic receptor to its ligands. Membrane hyperpolarization increases response to the full agonist acetylcholine (ACh) while decreasing response to the partial agonist pilocarpine. We previously have demonstrated that the regulator of G-protein signaling (RGS) 4 protein discriminates between the voltage-dependent responses of ACh and pilocarpine; however, the underlying mechanism remains unclear. Here we show that RGS4 is involved in the voltage-dependent behavior of the M2 muscarinic receptor-mediated signaling in response to pilocarpine. Additionally we revealed structural determinants on the M2 muscarinic receptor underlying the voltage-dependent response. By electrophysiological recording in Xenopus oocytes expressing M2 muscarinic receptor and G-protein-gated inwardly rectifying K+ channels, we quantified voltage-dependent desensitization of pilocarpine-induced current in the presence or absence of RGS4. Hyperpolarization-induced desensitization of the current required for RGS4, also depended on pilocarpine concentration. Mutations of charged residues in the aspartic acid-arginine-tyrosine motif of the M2 muscarinic receptor, but not intracellular loop 3, significantly impaired the voltage-dependence of RGS4 function. Thus, our results demonstrated that voltage-dependence of RGS4 modulation is derived from the M2 muscarinic receptor. These results provide novel insights into how membrane potential impacts G-protein signaling by modulating GPCR communication with downstream effectors.


Respiratory viruses in transplant recipients: more than just a cold. Clinical syndromes and infection prevention principles.
The aim of this review is to provide updated information on the clinical spectrum, treatment options, and infection prevention strategies for respiratory viral infections (RVIs) in both solid organ (SOT) and hematopoietic stem cell transplant (HSCT) patients. The MEDLINE and PubMed databases were searched for literature regarding the aforementioned aspects of RVIs, with focus on respiratory syncytial virus, adenovirus, influenza virus, parainfluenza virus, human metapneumovirus, and rhinovirus. Compared to immunocompetent hosts, SOT and HSCT patients are much more likely to experience a prolonged duration of illness, prolonged shedding, and progression of upper respiratory tract disease to pneumonia when infected with respiratory viruses. Adenovirus and respiratory syncytial virus tend to have the highest mortality and risk for disseminated disease, but all the RVIs are associated with higher morbidity and mortality in these patients than in the general population. These viruses are spread via direct contact and aerosolized droplets, and nosocomial spread has been reported. RVIs are associated with high morbidity and mortality among SOT and HSCT recipients. Management options are currently limited or lack strong clinical evidence. As community and nosocomial spread has been reported for all reviewed RVIs, strict adherence to infection control measures is key to preventing outbreaks.


Management of lacerated and swollen tongue after convulsive seizure with a mouth protector: interprofessional collaboration including dentists in intensive care.
We encountered a 74-year-old male patient with tongue laceration after convulsive seizures under intensive care. The tongue showed severe swelling, and the right ventral surface had been lacerated by his isolated and pointed right lower canine. Our university hospital has established a perioperative management center, and is promoting interprofessional collaboration, including dentists, in perioperative management. Dentists collaborating in the perioperative management center took dental impressions, with the support of anesthesiologists who opened the patient's jaw under propofol sedation, to produce a mouth protector. By raising the patient's bite, the completed mouth protector prevented the isolated tooth from contacting the tongue and protected the lacerated wound. Use of the mouth protector prevented the lacerated tongue from coming into contact with the pointed tooth, and the tongue healed gradually. These findings underscore that interprofessional collaboration including dentists can improve the quality of medical care.


Ultrasound-Guided Ethanol Ablation as a Primary Treatment for Thyroglossal Duct Cyst: Feasibility, Characteristics, and Outcomes.
To evaluate the feasibility, characteristics, and outcomes of ultrasound-guided ethanol ablation (US-EA) as a primary treatment for thyroglossal duct cysts (TGDCs). Prospective case series. Single center study. The inclusion criteria were as follows: (i) patients with TGDC aged ≥18 years, (ii) benign TGDC in imaging and cytological examinations, and (iii) patients' need for nonsurgical scarless treatment. US-EA was used as the primary treatment strategy. The primary outcome variables were the volume reduction rate (VRR) and cosmetic score at the last follow-up. We enrolled 28 patients with TGDC. The median TGDC volume at baseline was 6.7 mL. The median procedure time of the US-EA was 6.5 minutes. The median volumes of the cyst aspirate and injected ethanol were 4.0 and 2.0 mL, respectively. Overall, 18, 8, and 2 patients underwent 1, 2, and 3 treatment sessions, respectively. There were no complications. The median VRR was 96.2%, and the treatment success rate was 96.4%. The World Health Organization cosmetic score decreased from 4 (baseline) to 1 (after treatment) in all patients. The subjective grade for cosmetic satisfaction was satisfactory or highly satisfactory in all patients. The VRR, treatment success rate, and the number of treatment sessions did not differ as functions of the characteristics of the TGDC, including the initial volume, septation, debris, or viscosity of the cyst fluid. US-EA was feasible, safe, and effective in patients with TGDC. Therefore, US-EA can be used as a primary treatment for TGDC, evading general anesthesia and surgical scar.


Overcoming Fear, Uncertainty, and Doubt: Artificial Intelligence (AI) and the Value of Trust.
This is an editorial based on personal experience dealing with the fear, uncertainty, and doubt regarding artificial intelligence (AI) and radiology (my field of specialization). In the end, the most important tools to engage with these are education, research, and policy or regulation with the ultimate goal of forging trust, not just in the AI but also in the people that help make this technology possible.


Generation of Enterobacter sp. YSU auxotrophs using transposon mutagenesis.
Prototrophic bacteria grow on M-9 minimal salts medium supplemented with glucose (M-9 medium), which is used as a carbon and energy source. Auxotrophs can be generated using a transposome. The commercially available, Tn5-derived transposome used in this protocol consists of a linear segment of DNA containing an R6Kγ replication origin, a gene for kanamycin resistance and two mosaic sequence ends, which serve as transposase binding sites. The transposome, provided as a DNA/transposase protein complex, is introduced by electroporation into the prototrophic strain, Enterobacter sp. YSU, and randomly incorporates itself into this host's genome. Transformants are replica plated onto Luria-Bertani agar plates containing kanamycin, (LB-kan) and onto M-9 medium agar plates containing kanamycin (M-9-kan). The transformants that grow on LB-kan plates but not on M-9-kan plates are considered to be auxotrophs. Purified genomic DNA from an auxotroph is partially digested, ligated and transformed into a pir+ Escherichia coli (E. coli) strain. The R6Kγ replication origin allows the plasmid to replicate in pir+ E. coli strains, and the kanamycin resistance marker allows for plasmid selection. Each transformant possesses a new plasmid containing the transposon flanked by the interrupted chromosomal region. Sanger sequencing and the Basic Local Alignment Search Tool (BLAST) suggest a putative identity of the interrupted gene. There are three advantages to using this transposome mutagenesis strategy. First, it does not rely on the expression of a transposase gene by the host. Second, the transposome is introduced into the target host by electroporation, rather than by conjugation or by transduction and therefore is more efficient. Third, the R6Kγ replication origin makes it easy to identify the mutated gene which is partially recovered in a recombinant plasmid. This technique can be used to investigate the genes involved in other characteristics of Enterobacter sp. YSU or of a wider variety of bacterial strains.


Functional dissection of the conjugative coupling protein TrwB.
The conjugative coupling protein TrwB is responsible for connecting the relaxosome to the type IV secretion system during conjugative DNA transfer of plasmid R388. It is directly involved in transport of the relaxase TrwC, and it displays an ATPase activity probably involved in DNA pumping. We designed a conjugation assay in which the frequency of DNA transfer is directly proportional to the amount of TrwB. A collection of point mutants was constructed in the TrwB cytoplasmic domain on the basis of the crystal structure of TrwB Delta N70, targeting the nucleotide triphosphate (NTP)-binding region, the cytoplasmic surface, or the internal channel in the hexamer. An additional set of transfer-deficient mutants was obtained by random mutagenesis. Most mutants were impaired in both DNA and protein transport. We found that the integrity of the nucleotide binding domain is absolutely required for TrwB function, which is also involved in monomer-monomer interactions. Polar residues surrounding the entrance and inside the internal channel were important for TrwB function and may be involved in interactions with the relaxosomal components. Finally, the N-terminal transmembrane domain of TrwB was subjected to random mutagenesis followed by a two-hybrid screen for mutants showing enhanced protein-protein interactions with the related TrwE protein of Bartonella tribocorum. Several point mutants were obtained with mutations in the transmembranal helices: specifically, one proline from each protein may be the key residue involved in the interaction of the coupling protein with the type IV secretion apparatus.


Quantitative analogy between polymer-grafted nanoparticles and patchy particles.
We establish a quantitative analogy between polymer grafted nanoparticles (PGNPs) and patchy nanoparticles (NPs). Over much of the experimentally relevant parameter space, we show that PGNPs behave quantitatively like Janus NPs, with the patch size having a universal dependence on the number of grafts and the ratio of the size of the NPs to the grafted chain size. The widely observed anisotropic self-assembly of PGNP into superstructures can thus be understood through simple geometric considerations of single patch models, in the same spirit as the geometry-based surfactant models of Israelachvili.


Surface changes in the North Atlantic meridional overturning circulation during the last millennium.
Despite numerous investigations, the dynamical origins of the Medieval Climate Anomaly and the Little Ice Age remain uncertain. A major unresolved issue relating to internal climate dynamics is the mode and tempo of Atlantic meridional overturning circulation variability, and the significance of decadal-to-centennial scale changes in Atlantic meridional overturning circulation strength in regulating the climate of the last millennium. Here we use the time-constrained high-resolution local radiocarbon reservoir age offset derived from an absolutely dated annually resolved shell chronology spanning the past 1,350 years, to reconstruct changes in surface ocean circulation and climate. The water mass tracer data presented here from the North Icelandic shelf, combined with previously published data from the Arctic and subtropical Atlantic, show that surface Atlantic meridional overturning circulation dynamics likely amplified the relatively warm conditions during the Medieval Climate Anomaly and the relatively cool conditions during the Little Ice Age within the North Atlantic sector.


Current Perspectives on B Lymphocytes in the Immunobiology of Hepatocellular Carcinoma.
Immune cells infiltrating tumors are capable of significantly impacting carcinogenesis through cancer promotion and anticancer responses. There are many aspects of hepatocellular carcinoma (HCC) related T lymphocytes that are undergoing extensive studies, whereas the effect exerted by B lymphocytes remains a less researched area. In this study, the latest research on the effect of B lymphocytes as they infiltrate tumors in relation to HCC is presented. Their prognosis-related importance is analyzed, along with their function in the tumor microenvironment (TME), as well as the way that B cell biology can be employed to help create a B cell therapy strategy for HCC.


The optic nerve head in myocilin glaucoma.
Approximately 1 in 30 unselected patients with open-angle glaucoma (OAG) have a mutation in the myocilin gene. The purpose of this study was to describe the morphologic features of the optic nerve head (ONH) in myocilin glaucoma. A case-control design was adopted. Sixty-six patients heterozygous for a range of myocilin mutation (cases) were matched in disease severity to 105 patients with OAG known not to have a myocilin mutation (controls), using visual field findings. Quantifiable analysis of the ONH was undertaken of stereoscopic photographs, by using custom software with a z-screen. Subjective grading of the cup depth, lamina cribrosa pore shape and orientation, and the slope of the neuroretinal rim was performed by an examiner masked to the subject's mutation status. Mutation screening was conducted using either direct sequencing or single-stranded conformation polymorphism analysis. Patients with a myocilin mutation had glaucoma diagnosed earlier (P < 0.001) and had higher maximum recorded intraocular pressures (P < 0.001) than did the control OAG subjects. There was no significant (P > 0.05) difference in global disc area, global neuroretinal rim area, alpha-parapapillary atrophy, beta-parapapillary atrophy, slope of neuroretinal rim, or visible lamina cribrosa morphology between myocilin mutation carriers and patients with nonmyocilin glaucoma. Disc hemorrhages were identified more frequently in those without mutations (14/209 vs. 1/129), though this was not significant after correction for multiple hypothesis testing. No major structural or morphologic difference of the ONH was detected in pooled data from subjects who had myocilin mutations compared with data from individuals with nonmyocilin glaucoma.


Developing antiracist integrated health professionals.
As the Collaborative Family Healthcare Association (CFHA) has increased its focus on justice, diversity, and inclusion in integrated care delivery, it is increasingly evident that widespread upstream transformation is needed to ensure that the integrated care workforce is appropriately prepared to deliver equitable care. This column highlights the need for systemic change in admissions standards, integrated care curricula, student mentorship/sponsorship, and faculty development within higher education in order to support the success of Black, Indigenous, and People of Color (BIPOC) students and increase antiracist approaches among health care professionals. CFHA members working in, or in collaboration with, academia are uniquely poised to influence higher education systems to promote diversity, inclusion, and antiracism among future integrated health professionals. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


A microsatellite guided insight into the genetic status of adi, an isolated hunting-gathering tribe of northeast India.
Tibeto-Burman populations of India provide an insight into the peopling of India and aid in understanding their genetic relationship with populations of East, South and Southeast Asia. The study investigates the genetic status of one such Tibeto-Burman group, Adi of Arunachal Pradesh based on 15 autosomal microsatellite markers. Further the study examines, based on 9 common microsatellite loci, the genetic relationship of Adi with 16 other Tibeto-Burman speakers of India and 28 neighboring populations of East and Southeast Asia. Overall, the results support the recent formation of the Adi sub-tribes from a putative ancestral group and reveal that geographic contiguity is a major influencing factor of the genetic affinity among the Tibeto-Burman populations of India.


Scaling mimesis: Morphometric and ecomorphological similarities in three sympatric plant-mimetic fish of the family Carangidae (Teleostei).
The mimetic juveniles of a number of carangid fish species resemble plant parts floating near the water surface, such as leaves, seeds and other plant debris. The present study is the first to verify the morphological similarities and ecomorphological relationships between three carangids (Oligoplites saurus, Oligoplites palometa and Trachinotus falcatus) and their associated plant models. Behavioral observations were conducted in the estuary of Curuçá River, in northeastern Pará (Brazil) between August 2015 and July 2016. Individual fishes and associated floating objects (models) were sampled for comparative analysis using both geometric and morphometric approaches. While the mimetic fish and their models retain their own distinct, intrinsic morphological features, a high degree of morphological similarity was found between each fish species and its model. The morphometric analyses revealed a general tendency of isometric development in all three fish species, probably related to their pelagic habitats, during all ontogenetic stages.


Topical phenytoin for the treatment of neuropathic pain.
We developed and tested a new putative analgesic cream, based on the anticonvulsant phenytoin in patients suffering from treatment refractory neuropathic pain. The use of commercial topical analgesics is not widespread due to the facts that capsaicin creams or patches can give rise to side effects, such as burning, and analgesic patches (e.g., lidocaine 5% patches) have complex handling, especially for geriatric patients. Only in a few countries, compounded creams based on tricyclic antidepressants or other (co-)analgesics are available. Such topical analgesic creams, however, are easy to administer and have a low propensity for inducing side effects. We, therefore, developed a new topical cream based on 5% and 10% phenytoin and described three successfully treated patients suffering from neuropathic pain. All patients were refractory to a number of other analgesics. In all patients, phenytoin cream was effective in reducing pain completely, without any side effects, and the tolerability was excellent. The onset of action of the phenytoin creams was within 30 minutes. Phenytoin cream might become a new treatment modality of the treatment of neuropathic pain.


Reliability of Genetic Alterations in Predicting Ceftriaxone Resistance in Neisseria gonorrhoeae Globally.
Antimicrobial resistance in N. gonorrhoeae is increasing globally, and ceftriaxone is the recommended treatment for empirical therapy in most settings. Developing molecular assays to detect decreased ceftriaxone susceptibility is critical. Using PathogenWatch, a public database of N. gonorrhoeae genomes, antibiotic susceptibility data and DNA sequences of different genes associated with ceftriaxone resistance were extracted. That information was used to determine the sensitivity and specificity of different molecular markers and algorithms to predict decreased susceptibility to ceftriaxone. A total of 12,943 N. gonorrhoeae genomes were extracted from the PathogenWatch database, of which 9,540 genomes were used in the analysis. The sensitivity and specificity of specific molecular markers and algorithms were largely consistent with prior reports. Small variation (<10%) in either sensitivity or specificity occurred. Certain algorithms using different molecular markers at various prevalence of decreased ceftriaxone susceptibility identified a potentially clinically useful range of positive and negative predictive values. We validated previously described mutations and algorithms in a large public database containing a global collection of N. gonorrhoeae genomes. Certain mutations and algorithms resulted in sensitivity and specificity values consistent with those of prior studies. Further research is needed to integrate these markers and algorithms into the development of molecular assays to predict decreased ceftriaxone susceptibility. IMPORTANCE Antimicrobial resistance in Neisseria gonorrhoeae (N. gonorrhoeae), the causative agent of gonorrhea, is rising globally. Ceftriaxone is the last remaining antibiotic for empirical treatment of gonorrhea. Developing molecular tests to predict ceftriaxone resistance can help to improve detection and surveillance of ceftriaxone resistance. Here, we utilized PathogenWatch, a public global online database of N. gonorrhoeae genomes, to evaluate different genetic markers in predicting decreased susceptibility to ceftriaxone. We compiled MICs for ceftriaxone from the PathogenWatch database and used a computational approach to extract all the genetic markers from the genomic data. We determined the sensitivity and specificity for predicting decreased ceftriaxone susceptibility among several combinations of genetic markers. We identified several combinations of genetic markers with high predictive values for decreased susceptibility to ceftriaxone. These combinations of genetic markers might be promising candidates for future molecular tests to predict ceftriaxone resistance.


Inclusion of the fitness sharing technique in an evolutionary algorithm to analyze the fitness landscape of the genetic code adaptability.
The canonical code, although prevailing in complex genomes, is not universal. It was shown the canonical genetic code superior robustness compared to random codes, but it is not clearly determined how it evolved towards its current form. The error minimization theory considers the minimization of point mutation adverse effect as the main selection factor in the evolution of the code. We have used simulated evolution in a computer to search for optimized codes, which helps to obtain information about the optimization level of the canonical code in its evolution. A genetic algorithm searches for efficient codes in a fitness landscape that corresponds with the adaptability of possible hypothetical genetic codes. The lower the effects of errors or mutations in the codon bases of a hypothetical code, the more efficient or optimal is that code. The inclusion of the fitness sharing technique in the evolutionary algorithm allows the extent to which the canonical genetic code is in an area corresponding to a deep local minimum to be easily determined, even in the high dimensional spaces considered. The analyses show that the canonical code is not in a deep local minimum and that the fitness landscape is not a multimodal fitness landscape with deep and separated peaks. Moreover, the canonical code is clearly far away from the areas of higher fitness in the landscape. Given the non-presence of deep local minima in the landscape, although the code could evolve and different forces could shape its structure, the fitness landscape nature considered in the error minimization theory does not explain why the canonical code ended its evolution in a location which is not an area of a localized deep minimum of the huge fitness landscape.


Protein kinase G regulates the basal tension and plays a major role in nitrovasodilator-induced relaxation of porcine coronary veins.
Coronary venous activity is modulated by endogenous and exogenous nitrovasodilators. The present study was to determine the role of protein kinase G (PKG) in the regulation of the basal tension and nitrovasodilator-induced relaxation of coronary veins. Effects of a PKG inhibitor on the basal tension and responses induced by nitroglycerin, DETA NONOate, and 8-Br-cGMP in isolated porcine coronary veins were determined. Cyclic cGMP was measured with radioimmunoassay. PKG activity was determined by measuring the incorporation of 32P from gamma-32P-ATP into the specific substrate BPDEtide. Rp-8-Br-PET-cGMPS, a specific PKG inhibitor, increased the basal tension of porcine coronary veins and decreased PKG activity. The increase in tension was 38% of that caused by nitro-L-arginine. Relaxation of the veins induced by nitroglycerin and DETA NONOate was accompanied with increases in cGMP content and PKG activity. These effects were largely eliminated by inhibiting soluble guanylyl cyclase with ODQ. The increase in PKG activity induced by the nitrovasodilators was abolished by Rp-8-Br-PET-cGMPS. The relaxation caused by these dilators and by 8-Br-cGMP at their EC50 was attenuated by the PKG inhibitor by 51-66%. These results suggest that PKG is critically involved in nitric oxide-mediated regulation of the basal tension in porcine coronary veins and that it plays a primary role in relaxation induced by nitrovasodilators. Since nitric oxide plays a key role in modulating coronary venous activity, augmentation of PKG may be a therapeutic target for improving coronary blood flow.


Early Helicobacter pylori colonisation: the association with growth faltering in The Gambia.
Helicobacter pylori is one of the commonest causes of chronic infection of mankind, yet the natural history of acute infection is poorly understood. Some studies suggest that gastric colonisation with H pylori is associated with suboptimal nutrition and growth in childhood. To describe the clinical features of early H pylori colonisation and assess its role in the development of infant malnutrition and growth faltering. Two consecutive prospective longitudinal cohort studies were conducted at the Medical Research Council Laboratories in a rural community in The Gambia, West Africa. The first birth cohort of 125 infants was followed by a second of 65 children from the same community. H pylori colonisation was detected by sequential 13C urea breath tests, and infant growth was monitored by serial measurements. Children with early H pylori colonisation became significantly lighter, shorter, and thinner than their peers in late infancy. The association was found in both cohorts. No socioeconomic or demographic confounding variables were identified to explain this, and the weight deficit was no longer detectable when the children were aged 5-8 years. Results suggest that H pylori colonisation in early infancy predisposes to the development of malnutrition and growth faltering, although the effect did not persist into later childhood.


Seasonal Migration of Pantala flavescens Across the Bohai Strait in Northern China.
Pantala flavescens (Fabricius 1798) (Odonata: Libellulidae) is one of the most common species of migratory dragonflies. P. flavescens adults were captured by a searchlight trap on Beihuang Island (BH Island; 38°24'N, 120°55'E) from 2003 to 2016, where there is no freshwater. This inspired our research to analyze the pattern of seasonal migration and population dynamics. Stable hydrogen isotope measurement and the Hybrid Single Particle Lagrangian Integrated Trajectory (HYSPLIT) were used to simulate the migration pathway of P. flavescens between different breeding habitats. The results showed that there was no significant difference among population numbers of this overseas migration across years (F13, 2161 = 0.85, P = 0.604); however, the numbers were significantly different across months (F5, 2161 = 3.91, P = 0.003). Our geospatial natal assignment model suggested that P. flavescens trapped on BH were originated in different geographical regions and might have three movement strategies: wandering around northern China and north-bound (positive) and south-bound (negative) movements. Among them, the majority were engaged in wandering around northern China. Model simulations suggested that P. flavescens toured around BH. The results contribute to the knowledge of P. flavescens population ecology in a large-scale geographic region and will aid in the prediction and interpretation of insect migration patterns in response to climate change.


Suicidal ideation in university students and other groups.
A questionnaire was used to investigate the amount of suicidal ideation in a sample of University students and two control groups from general practice. A high proportion of students (55%) and of the General Practice group (45%) admitted to some suicidal ideation in their life time. The study confirmed the excess of females over males concerning all aspects of suicidal ideation and behaviour. The significance of these findings is discussed.


Multicentre survey of the in-vitro activity of seven antimicrobial agents, including ertapenem, against recently isolated Gram-negative anaerobic bacteria in Greece.
The in-vitro activities of penicillin, ticarcillin-clavulanic acid, cefoxitin, imipenem, ertapenem, metronidazole and clindamycin were evaluated against 138 Gram-negative anaerobic isolates (82 Bacteroides fragilis group, 17 non-fragilis Bacteroides spp., 31 Prevotella spp., four Fusobacterium spp., two Veillonella spp., one Porphyromonas sp. and one Tissierella praeacuta) collected from six general hospitals in Athens, Greece. Overall rates of non-susceptibility (both resistant and intermediately-resistant) to penicillin and ticarcillin-clavulanic acid were 81.8% and 2.3%, respectively. The rates of non-susceptibility to cefoxitin and clindamycin were 30.3% and 31.1%, respectively, and that for metronidazole was 4.3% (four Prevotella spp. isolates, one Porphyromonas sp. isolate and one B. fragilis isolate). Only the single B. fragilis isolate was nim-positive by PCR. Only one B. fragilis isolate was resistant to both carbapenems tested, while six more Bacteroides spp. isolates were imipenem-susceptible and ertapenem-non-susceptible. The MIC range, MIC(50) and MIC(90) values were comparable for imipenem and ertapenem, although ertapenem MIC(90)s were one or two two-fold dilutions higher.


Autoantibodies specific for the cardiac myosin isoform are found in mice susceptible to Coxsackievirus B3-induced myocarditis.
Several mouse strains are susceptible to immunopathic myocarditis after infection with Coxsackievirus B3 (CB3). This disease is associated with autoantibodies that are directed against myosin. In this study we characterized sera from CB3-infected mice for their reactivity with three different myosin isoforms (heart, skeletal muscle, and brain myosins) and for autoantibody isotype by using an ELISA. Competitive inhibition assays and absorption studies with various myosins demonstrated the presence of two autoantibody populations in sera of susceptible A.CA and A.SW mice. The first was specific for cardiac myosin and was mainly IgG. The second antibody population cross-reacted with heart, skeletal muscle, and brain myosin and was mainly IgM. B10.PL/SgSf and B10.A/SgSf mice, which do not develop immunopathic myocarditis, produced only the IgM autoantibody population cross-reactive with all three myosin isoforms. Because the heart-specific myosin autoantibodies were found exclusively in the mouse strains that developed immunopathic myocarditis, they can be considered a serologic marker for autoimmune heart disease.


Cartilage oligomeric matrix protein-deficient mice have normal skeletal development.
Cartilage oligomeric matrix protein (COMP) belongs to the thrombospondin family and is a homopentamer primarily expressed in cartilage. Mutations in the COMP gene result in the autosomal dominant chondrodysplasias pseudoachondroplasia (PSACH) and some types of multiple epiphyseal dysplasia (MED), which are characterized by mild to severe short-limb dwarfism and early-onset osteoarthritis. We have generated COMP-null mice to study the role of COMP in vivo. These mice show no anatomical, histological, or ultrastructural abnormalities and show none of the clinical signs of PSACH or MED. Northern blot analysis and immunohistochemical analysis of cartilage indicate that the lack of COMP is not compensated for by any other member of the thrombospondin family. The results also show that the phenotype in PSACH/MED cartilage disorders is not caused by the reduced amount of COMP.


Stereoselective syntheses of (+/-)-komaroviquinone and (+/-)-faveline methyl ether through intramolecular Heck reaction.
[reaction: see text] An efficient, flexible, and stereoselective convergent route for constructing the trans-10-hydroxy-1,1-dimethyloctahydrodibenzo[a,d]cyclohepten-7-ones (5a-c) was achieved via intramolecular Heck reaction. This strategy has been successfully implemented for the syntheses of (+/-)-komaroviquinone (3) through (+/-)-coulterone dimethyl ether (5c) and (+/-)-faveline methyl ether (1a).


Is prehypertension more strongly associated with long-term ambient air pollution exposure than hypertension? Findings from the 33 Communities Chinese Health Study.
Numerous studies have evaluated the effects of long-term exposure to ambient air pollution on hypertension. However, little information exists regarding its effects on prehypertension, a very common, but understudied cardiovascular indicator. We evaluated data from 24,845 adults (ages 18-74 years) living in three Northeastern Chinese cities in 2009. Blood pressure (BP) was measured by trained observers using a standardized mercuric-column sphygmomanometer. Three-year (from 2006 to 2008) average concentrations of particles with an aerodynamic diameter ≤10 μm (PM10), sulfur dioxide (SO2), nitrogen dioxides (NO2), and ozone (O3) were calculated using data from monitoring stations. Effects were analyzed using generalized additive models and two-level regression analyses, controlling for covariates. We found positive associations of all pollutants with prehypertension (e.g. odds ratio (OR) was 1.17 (95% confidence interval (CI), 1.09-1.25) per interquartile range (IQR) of PM10) in a fully adjusted model, as compared to normotensive participants. These associations were stronger than associations with hypertension (e.g. OR was 1.03 (95% CI, 1.00, 1.07) per IQR of PM10). We have also found positive associations of all studied pollutants with systolic and diastolic BP: e.g., associations with PM10 per IQR were 1.24 mmHg (95% CI, 1.03-1.45) for systolic BP and 0.47 mmHg (95% CI, 0.33-0.61) for diastolic BP. Further, we observed that associations with BP were stronger in women and in older participants (systolic BP only). In conclusion, long-term exposure to ambient air pollution was more strongly associated with prehypertension than with hypertension, especially among females and the elderly. Thus, interventions to reduce air pollution are of great significance for preventing future cardiovascular events, particularly among individuals with prehypertension.


Using fluorescent lipids in live zebrafish larvae: From imaging whole animal physiology to subcellular lipid trafficking.
Lipids serve essential functions in cells as signaling molecules, membrane components, and sources of energy. Defects in lipid metabolism are implicated in a number of pandemic human diseases, including diabetes, obesity, and hypercholesterolemia. Many aspects of how fatty acids and cholesterol are absorbed and processed by intestinal cells remain unclear and present a hurdle to developing approaches for disease prevention and treatment. Numerous studies have shown that the zebrafish is an excellent model for vertebrate lipid metabolism. In this chapter, we review commercially available fluorescent lipids that can be deployed in live zebrafish to better understand lipid signaling and metabolism. In this chapter, we present criteria one should consider when selecting specific fluorescent lipids for the study of digestive physiology or lipid metabolism in larval zebrafish.


Motor vehicle restraint system use and risk of spine injury.
Motor vehicle collision (MVC)-related spinal injury is a severe and often permanently disabling injury. In addition, strain injuries have been reported as a common outcome of MVCs. Although advances in automobile crashworthiness have reduced both fatalities and severe injuries, the impact of varying occupant restraint systems (seatbelts and airbags) on thoracolumbar spine injuries is unknown. This study examined the relationship between the occurrence of mild to severe cervical and thoracolumbar spine injury and occupant restraint systems among front seat occupants involved in frontal MVCs. A retrospective cohort study was conducted among subjects obtained from the 1995-2004 National Automotive Sampling System. Cases were identified based on having sustained a spine injury of >/=1 on the Abbreviated Injury Scale (AIS), 1990 Revision. Risk risks (RRs) and 95% confidence intervals (CIs) were computed comparing occupant restraint systems with unrestrained occupants. We found an overall incidence of AIS1 cervical (11.8%) and thoracolumbar (3.7%) spinal injury. Seatbelt only restraints were associated with increased cervical AIS1 injury (RR = 1.40, 95% CI 1.04-1.88). However, seatbelt only restraints showed the greatest risk reduction for AIS2 spinal injuries. Airbag only restraints reduced thoracolumbar AIS1 injuries (RR = 0.29, 95% CI 0.08-1.04). Seatbelt combined with airbag use was protective for cervical AIS3+ injury overall (RR = 0.29, 95% CI 0.14-0.58), cervical neurological injury (RR = 0.19, 95% CI 0.05-0.81), and thoracolumbar AIS3+ injury overall (RR = 0.20, 95% CI 0.05-0.70). The results of this study suggest that seatbelts alone or in combination with an airbag increased the incidence of AIS1 spinal injuries, but provide protection against more severe injury to all regions of the spine. Airbag deployment without seatbelt use did not show increased protection relative to unrestrained occupants.


Vectors expressing chimeric Japanese encephalitis dengue 2 viruses.
Vectors based on self-replicating RNAs (replicons) of flaviviruses are becoming powerful tool for expression of heterologous genes in mammalian cells and development of novel antiviral and anticancer vaccines. We constructed two vectors expressing chimeric viruses consisting of attenuated SA14-14-2 strain of Japanese encephalitis virus (JEV) in which the PrM/M-E genes were replaced fully or partially with those of dengue 2 virus (DENV-2). These vectors, named pJED2 and pJED2-1770 were transfected to BHK-21 cells and produced chimeric viruses JED2V and JED2-1770V, respectively. The chimeric viruses could be passaged in C6/36 but not BHK-21 cells. The chimeric viruses produced in C6/36 cells CPE 4-5 days after infection and RT-PCR, sequencing, immunofluorescence assay (IFA) and Western blot analysis confirmed the chimeric nature of produced viruses. The immunogenicity of chimeric viruses in mice was proved by detecting DENV-2 E protein-specific serum IgG antibodies with neutralization titer of 10. Successful preparation of infectious clones of chimeric JEV-DENV-2 viruses showed that JEV-based expression vectors are fully functional.


Protein Supplementation Does Not Further Increase Latissimus Dorsi Muscle Fiber Hypertrophy after Eight Weeks of Resistance Training in Novice Subjects, but Partially Counteracts the Fast-to-Slow Muscle Fiber Transition.
The response to resistance training and protein supplementation in the latissimus dorsi muscle (LDM) has never been investigated. We investigated the effects of resistance training (RT) and protein supplementation on muscle mass, strength, and fiber characteristics of the LDM. Eighteen healthy young subjects were randomly assigned to a progressive eight-week RT program with a normal protein diet (NP) or high protein diet (HP) (NP 0.85 vs. HP 1.8 g of protein·kg(-1)·day(-1)). One repetition maximum tests, magnetic resonance imaging for cross-sectional muscle area (CSA), body composition, and single muscle fibers mechanical and phenotype characteristics were measured. RT induced a significant gain in strength (+17%, p < 0.0001), whole muscle CSA (p = 0.024), and single muscle fibers CSA (p < 0.05) of LDM in all subjects. Fiber isometric force increased in proportion to CSA (+22%, p < 0.005) and thus no change in specific tension occurred. A significant transition from 2X to 2A myosin expression was induced by training. The protein supplementation showed no significant effects on all measured outcomes except for a smaller reduction of 2X myosin expression. Our results suggest that in LDM protein supplementation does not further enhance RT-induced muscle fiber hypertrophy nor influence mechanic muscle fiber characteristics but partially counteracts the fast-to-slow fiber shift.


Co-culture of extracellular matrix suppresses the cell death of rat pancreatic islets.
The purpose of this study was to investigate the effect of culture microenvironment on the cell death of isolated rat pancreatic islets. After isolation by the conventional collagenase digestion technique, the islets were cultured in a hydrogel of collagen type I mixed with collagen type III, type IV, and laminin. Irrespective of the type of mixture, islet cell death was significantly suppressed by their co-culture with the collagen hydrogel mixtures, although no change in islet morphology was observed. Co-culture with the collagen mixtures had no influence on the expressed mRNA level of insulin, glucagon, and somatostatin of the islets, or the islet secretion of hepatocyte growth factor (HGF), interleukin (IL)-1alpha, and IL-1beta. These findings suggest that three-dimensional culture in the collagen hydrogel and the mixture of laminin is able to maintain the cell viability of islets.


Attending a Biopsychosocially Focused Buprenorphine Training Improves Clinician Attitudes.
Objective: Substance use disorders remain highly stigmatized. Access to medications for opioid use disorder is poor. There are many barriers to expanding access including stigma and lack of medical education about substance use disorders. We enriched the existing, federally required, training for clinicians to prescribe buprenorphine with a biopsychosocial focus in order to decrease stigma and expand access to medications for opioid use disorder. Methods: We trained a family medicine team to deliver an enriched version of the existing buprenorphine waiver curriculum. The waiver training was integrated into the curriculum for all University of Rochester physician and nurse practitioner family medicine residents and also offered to University of Rochester residents and faculty in other disciplines and regionally. We used the Brief Substance Abuse Attitudes Survey to collect baseline and post-training data. Outcomes: 140 training participants completed attitude surveys. The overall attitude score increased significantly from pre to post-training. Additionally, significant changes were observed in non-moralism from pre-training (M = 20.07) to post-training (M = 20.98, p < 0.001); treatment optimism from pre-training (M = 21.56) to post-training (M = 22.33, p < 0.001); and treatment interventions from pre-training (M = 31.03) to post-training (M = 32.10, p < 0.001). Conclusion: Increasing medical education around Opioid Use Disorder using a Family Medicine trained team with a biopsychosocial focus can improve provider attitudes around substance use disorders. Enriching training with cases may improve treatment optimism and may help overcome the documented barriers to prescribing medications for opioid use disorder and increase access for patients to lifesaving treatments.