Serum albumin as a primary non-covalent binding protein for nitro-oleic acid.
This work explores the interaction of 9/10-nitro-oleic acid (NO2-OA) with human serum albumin (HSA). The molecular mechanism of the biological action of NO2-OA is to our knowledge based on a reversible covalent reaction-Michael addition of nucleophilic amino acid residues of proteins. Since HSA is an important fatty acid transporter, a key question is whether NO2-OA can bind covalently or non-covalently to HSA, similarly to oleic acid (OA), which can interact with the FA1-FA7 binding sites of the HSA molecule. 1H NMR studies and competition analysis with OA and the drugs ibuprofen and warfarin were used to investigate a potential non-covalent binding mode. NO2-OA/HSA binding was confirmed to compete with warfarin for FA-7 with significantly higher affinity. NO2-OA competes with ibuprofen for FA-3 and FA-6, however, in contrast to the situation with warfarin, the binding affinities are not significantly different. The described interactions are based exclusively on non-covalent binding. No covalent binding of NO2-OA to HSA was detected by MS/MS. More detailed studies based on MALDI-TOF-MS and Ellman's assay indicated that HSA can be covalently modified in the presence of NO2-OA to a very limited extent. It was also shown that NO2-OA has a higher affinity to HSA than that of OA.


Serum and sputum antibiotic levels after ampicillin, amoxycillin and bacampicillin chronic bronchitis patients.
Serum and sputum concentrations of ampicillin or amoxycillin were measured in patients admitted to hospital for acute exacerbations of chronic bronchitis with purulent sputum. Mean peak serum levels of nearly 12 mg/l were found after 1600 mg bacampicillin (mean peak level in sputum 0.85 mg/l). The serum and sputum concentrations after 750 mg amoxycillin and 800 mg bacampicillin were comparable (mean peak serum levels approximately 9.5 mg/l, sputum concentrations 0.4 to 0.5 mg/l) although the drugs were not given in equimolar doses. Results after 1000 mg ampicillin by mouth were less satisfactory (mean peak serum level 7.8 mg/l) and only 0.25 mg/l was attained in the sputum. Minimum inhibitory concentrations of ampicillin and amoxycillin were measured for 177 Haemophilus influenzae strains. Most of the ampicillin MIC values were between 0.125 and 0.5 mg/l but more of the strains required 0.5 mg/l of amoxycillin. The amoxycillin MIC values were often one or two dilutions higher than those of ampicillin (p less than 0.001).


A randomized trial comparing bipolar transurethral vaporization of the prostate with GreenLight laser (xps-180watt) photoselective vaporization of the prostate for treatment of small to moderate benign prostatic obstruction: outcomes after 2 years.
To test the non-inferiority of bipolar transurethral vaporization of the prostate (TUVP) compared to GreenLight laser (GL) photoselective vaporization of the prostate (PVP) for reduction of benign prostatic hyperplasia-related lower urinary tract symptoms in a randomized trial. Eligible patients with prostate volumes of 30-80 mL were randomly allocated to GL-PVP (n = 58) or bipolar TUVP (n = 61). Non-inferiority of symptom score (International Prostate Symptom Score [IPSS]) at 24 months was evaluated. All peri-operative variables were recorded and compared. Urinary (IPSS, maximum urinary flow rate and post-void residual urine volume) and sexual (International Index of Erectile Function-15) outcome measures were evaluated at 1, 4, 12 and 24 months. Need for retreatment and complications, change in PSA level and health resources-related costs of both procedures were recorded and compared. Baseline and peri-operative variables were similar in the two groups. At 1, 4, 12 and 24 months, 117, 116, 99 and 96 patients, respectively, were evaluable. Regarding urinary outcome measures, there was no significant difference between the groups. The mean ± sd IPSS at 1 and 2 years was 7.1 ± 3 and 7.9 ± 2.9 (P = 0.8), respectively, after GL-PVP and 6.3 ± 3.1 and 7.2 ± 2.8, respectively, after bipolar TUVP (P = 0.31). At 24 months, the mean difference in IPSS was 0.7 (95% confidence interval -0.6 to 2.3; P = 0.6). The median (range) postoperative PSA reduction was 64.7 (25-99)% and 65.9 (50-99)% (P = 0.006) after GL-PVP, and 32.1 (28.6-89.7)% and 39.3 (68.8-90.5)% (P = 0.005) after bipolar TUVP, at 1 and 2 years, respectively. After 2 years, retreatment for recurrent bladder outlet obstruction was reported in eight (13.8%) and 10 (16.4%) patients in the GL-PVP and bipolar TUVP groups, respectively (P = 0.8). The mean estimated cost per bipolar TUVP procedure was significantly lower than per GL-PVP procedure after 24 months (P = 0.01). In terms of symptom control, bipolar TUVP was not inferior to GL-PVP at 2 years. Durability of the outcome needs to be tracked. The greater cost of GL-PVP compared with bipolar TUVP is an important concern.


A tiered approach to optimize pediatric laparoscopic appendectomy outcomes.
Surgical management of appendicitis accounts for ~30% of total expenditure in the practice of pediatric surgery and is associated with high cost variation. We hypothesize that incorporating single-incision laparoscopy (SILS) and the resultant by-product dual-incision laparoscopy (DILS) into a historically three-incision laparoscopic (TILS) appendectomy practice affords equal outcomes at lower cost. Appendectomies performed at a large-volume tertiary care children's hospital from 1/2015-12/2017 were retrospectively reviewed. Appendectomy technique and appendicitis severity were stratified against operative and admission direct variable (DV) costs. Secondary outcomes included perioperative time course and 30-day postoperative outcomes. A total of 970 appendectomies were analyzed during the study period (61% acute, 39% complex appendicitis). SILS and DILS had significantly lower mean DV costs and OR times compared to TILS for both acute and complex appendicitis while maintaining equivalent outcomes. SILS and DILS appendectomy techniques can be incorporated into pediatric surgical practice at lower cost than TILS appendectomy while maintaining equivalent outcomes. Further, the introduction of a tiered approach to laparoscopic appendectomy, in which all cases are started as SILS with additional incisions added based on operative difficulty, is estimated to save $74,580 annually in operative DV costs at a pediatric surgical center averaging 314 laparoscopic appendectomies per year. Treatment Study. Level III.


Can Money Buy Happiness for a Family? Family Income, Parenting Style, and Life Satisfaction in Parents and Adolescents.
This study investigated the associations among family income and life satisfaction in parents, and parenting styles, income, and overall happiness and depression in their adolescents. A total of 1,913 participants comprising adolescents (aged 14-16) and their parents were recruited for this study. Participants were assessed using self-report questionnaires. Monthly household income was recorded by an open-ended question item. The life satisfaction of parents and overall happiness of adolescents were assessed by one item each, rated on a 4-point Likert scale. Three subtypes of parenting style (affectionate, monitoring, over-control) were evaluated by adolescents using an inventory consisting of 21 items on a 4-point Likert scale. Adolescents' depression was assessed with a checklist rated by the adolescents, consisting of 10 items on a 4-point Likert scale. The odds ratio (OR) for life dissatisfaction in adults was significantly higher in the lower (1st and 2nd) quintile groups of household income, by 9.94 (p < .001) and 3.50 (p < .001) folds, respectively, compared to the highest (5th) group. In contrast, household income had no significant association with life unhappiness in adolescents. Further, total household income had no significant association with the adolescent's depression score in linear regression analysis, but parenting style did. While affectionate (β = -0.260; p < .001) and monitoring (β = -0.100; p < .001) parenting styles showed negative associations with adolescents' depression scores, over-control style (β = 0.120; p < .001) showed a positive association. The results indicate the differences in factors impacting the life satisfaction of adults and happiness of adolescents, and the importance of effective parenting for adolescents' mental health.


Self-efficacy in health among university students: the role of social support and place.
The aim of this study was to better understand the role of students' living arrangements (on campus, off campus, with and without their parents) on their health self-efficacy. A sample of undergraduate students (n = 216) were recruited by using word of mouth and visiting classrooms in a small Atlantic Canadian university. Participants completed a self-report questionnaire measuring health and sleep self-efficacy. Analyses of variance and covariance were used to compare the living arrangements of the students. Differences between living arrangements were found. Students living on campus had higher health self-efficacy, particularly on the psychological well-being subscale, followed by those living off campus with their parents. Students living off campus without their parents had the lowest scores. This study demonstrated that health self-efficacy varies according to students' living situations, thus illustrating the importance of keeping students' living arrangements in mind when designing health and well-being interventions.


Anxiolytic effects of the novel α2δ ligand mirogabalin in a rat model of chronic constriction injury, an experimental model of neuropathic pain.
Psychiatric disorders such as anxiety and depression are frequently observed in neuropathic pain patients, and negatively impact their quality of life. Mirogabalin is a novel ligand for the α2δ subunit of voltage-gated calcium channels and has unique binding characteristics to α2δ subunits and potent and long-lasting analgesic effects in neuropathic pain models. To provide further information on the pharmacological profile of mirogabalin and its utility for chronic pain therapy, we investigated its anxiolytic effects in an experimental animal model for neuropathic pain. In chronic constriction injury (CCI) model rats, mechanical hypersensitivity was determined by the von Frey test. Anxiety- and depression-related behaviours were evaluated using the elevated plus maze test and forced swimming test, respectively. CCI model rats showed sustained tactile allodynia followed by anxiety-related behaviours, not depression-related behaviours. The tactile allodynia (significant decreases in paw withdrawal threshold) developed within 2 weeks after model preparation, whereas the anxiety-related behaviours (significant decreases in the number of entries and time spent in open arms and significant increases in time spent in closed arms) were observed at 5 weeks but not 4 weeks after model preparation. Single oral administration of mirogabalin (3 or 10 mg/kg) dose-dependently alleviated the above-mentioned anxiety-related behaviours and tactile allodynia. CCI model rats showed anxiety-related behaviours in a time-dependent manner in the elevated plus maze test. Mirogabalin alleviated both the anxiety-related behaviours and tactile allodynia in CCI model rats. Mirogabalin may provide effective anxiety relief as well as pain relief in patients with neuropathic pain.


Solid-state NMR and 2,3-dicyano-5,7-dimethyl-6H-1,4-diazepine.
Crystalline 2,3-dicyano-5,7-dimethyl-6H-1,4-diazepine (A) was investigated by solid-state NMR spectroscopy, X-ray diffraction, and spectral simulations. The solid-state 13C NMR spectra of A display peculiar splittings for the methyl and cyano resonances. The crystal structure of A indicates that the methyl doublet is a consequence of two crystallographically inequivalent environments. The methyl motions associated with each site was examined via spin-lattice relaxation time (T1) measurements, and the carbon relaxation times (T(1)C) were used to calculate energy barriers to methyl rotation. The energy barriers to rotation were then used to correlate each methyl 13C shift with a particular crystallographic environment. The complex cyano splittings, however, are a result of both crystallographic inequivalence and residual 13C-14N dipolar coupling. The multiplet patterns of the isotropic shifts (centerbands) are dependent upon the magic-angle spinning (MAS) rate. Spectral simulations, using the perturbation method, of the centerbands and first-order sidebands were used to demonstrate, and elucidate, the observed MAS rate-dependent multiplet patterns of the cyano signals.


Selection of fully processed HIV-1 nucleocapsid protein is required for optimal nucleic acid chaperone activity in reverse transcription.
The mature HIV-1 nucleocapsid protein (NCp7) is generated by sequential proteolytic cleavage of precursor proteins containing additional C-terminal peptides: NCp15 (NCp7-spacer peptide 2 (SP2)-p6); and NCp9 (NCp7-SP2). Here, we compare the nucleic acid chaperone activities of the three proteins, using reconstituted systems that model the annealing and elongation steps in tRNA(Lys3)-primed (-) strong-stop DNA synthesis and subsequent minus-strand transfer. The maximum levels of annealing are similar for all of the proteins, but there are important differences in their ability to facilitate reverse transcriptase (RT)-catalyzed DNA extension. Thus, at low concentrations, NCp9 has the greatest activity, but with increasing concentrations, DNA synthesis is significantly reduced. This finding reflects NCp9's strong nucleic acid binding affinity (associated with the highly basic SP2 domain) as well as its slow dissociation kinetics, which together limit the ability of RT to traverse the nucleic acid template. NCp15 has the poorest activity of the three proteins due to its acidic p6 domain. Indeed, mutants with alanine substitutions for the acidic residues in p6 have improved chaperone function. Collectively, these data can be correlated with the known biological properties of NCp9 and NCp15 mutant virions and help to explain why mature NC has evolved as the critical cofactor for efficient virus replication and long-term viral fitness.


[Treatment of voluminous keratocysts of the maxilla (author's transl)].
Recurrences of keratocysts are infrequently reported. Excision of these cysts should therefore always be conservative in nature, preferably by interrupting resection. Bimaxillary blocking is not essential, even in the most voluminous cysts.


Acute pancreatitis as an early indicator of pancreatic head carcinoma.
The purpose of this study was to determine if AP is an indicator of pancreatic head carcinoma, and if it is associated with disease stage. A retrospective review of the medical records of 154 patients with pancreatic head cancer who were treated from January 2006 to December 2011 was conducted. Patients were divided into 2 groups: those with AP and those without. Data extracted and compared including age, gender, the presence of AP, and laboratory results. This study included 103 males (67.8%) and 49 females (32.9%), with an age range of 38-83 years. The frequency of main pancreatic duct dilation (MPD) was higher in patients without pancreatitis than in patients with AP (P = 0.018). There were 18, 14, 4, and 2 patients in the AP group, and 34, 26, 31, and 23 patients in nonpancreatitis group with stage I, II, III, and IV disease, respectively, (P = 0.007). The median length of tumors in AP group was 3.41 cm, and that in the non-pancreatitis group was 4.15 cm (P = 0.028).No correlation was found between disease stage and the presence of MPD dilatation (P = 0.646). The area under the receiver operating characteristic curve (AUC) of 0.66 (P = 0.003; 95% confidence interval [CI] 0.568-0.753) indicated that AP itself was insufficient for predicting disease stage. AP is an early manifestation of pancreatic head carcinoma, although the presence of AP is insufficient to predict disease stage.


Influence of age and bladder dysfunction on the contractile properties of isolated human detrusor smooth muscle.
To test the hypothesis that the in vitro contractile properties of human detrusor smooth muscle are dependent on the age, gender and lower urinary tract pathology of the patient. Contractions were elicited in isolated human detrusor smooth muscle preparations by nerve-mediated electrical field stimulation, agonist application (carbachol, α,β-methylene ATP and high-K solutions) or direct muscle electrical stimulation. Biopsies (n = 227) were obtained from four groups of patients with: stable bladders (control), bladder outlet obstruction (BOO), idiopathic (IDO), or neurogenic (NDO) detrusor overactivity. The magnitude of nerve-mediated contractions declined as a function of patients' age in each of the BOO, IDO and NDO groups but not in the control group. Contractions elicited by direct muscle activation (10 µM carbachol or electrical stimulation with 20 ms pulses in the presence of 1 µM tetrodotoxin) did not vary with patient age. Carbachol contractions were significantly smaller in samples from NDO bladders. Atropine resistance was more prevalent in the pathology groups compared with the control group and was greatest in the IDO group. There was no influence of age in the prevalence or magnitude of atropine-resistant contractions in any group. Muscle excitability to direct electrical stimulation was similar in all groups. In the human bladder there is no evidence for a decline of detrusor smooth muscle contractility or excitability as a function of age, nor any gender difference or presence of pathology. In the pathology groups there was evidence for a decline of functional innervation with age.


Prognostic value of oncoprotein expressions in thyroid papillary carcinoma.
The aim of this study was to evaluate the expressions of oncoproteins and to correlate the results with clinicopathologic parameters in papillary thyroid carcinoma (PTC). Papillary thyroid cancer (PTC) is the most common form and accounts for about 80% of all thyroid cancers. Although PTC generally has a good prognosis, some patients suffer from local recurrence and/or distant metastasis. Oncogenes have reported to be related not only in carcinogenesis but also in tumor prognosis, tumor type, differentiation and site of tumor in epithelial malignant tumors such as thyroid, breast, ovarian, and stomach cancer. This study was planned retrospectively and was performed in 87 patients (47 PTC, 40 benign lesions). The data of clinicopathologic parameters and tissue samples were collected from the archives. Sections stained with H&E were evaluated for each case and after confirming the diagnosis of PTC, oncoprotein expressions were determined by immunohistochemical analysis. The differences of oncoprotein expressions in PTC compared with control group were statistically significant. Cyclin D1 and p53 expressions were significantly increased in PTC. The expressions of bcl-2 and c-erbB-2 in PTC were found as increased, but the correlation between these proteins and poor prognostic parameters were not significant. We suggest that increased expressions of cyclin D1 and p53 could be used as prognostic factors in patients with PTC.


Species delimitation of Margattea cockroaches from China, with seven new species (Blattodea, Ectobiidae, Pseudophyllodromiinae).
Nearly 450 Margattea specimens were collected from 27 locations in China and their morphology was examined. Then 68 Margattea COI sequences were obtained and used to carry out phylogenetic analyses as well as species delimitation analyses using General Mixed Yule Coalescent (GMYC), Automatic Barcode Gap Discovery (ABGD), and Poisson Tree Processes (bPTP). GMYC analysis resulted in 21 molecular operational taxonomic units (MOTUs) (confidence interval: 20-22), which was completely consistent with the result of the bPTP. There were 15 MOTUs using the ABGD method. The number of MOTUs was slightly different from the assigned morphospecies (16). As to the incongruence between molecular and morphological results, we checked the specimens again and made sure that most morphological differences were determined to be intraspecific differences (except the difference between M. angusta and M. mckittrickae), although a large genetic distance existed. Finally, 16 Margattea species from China were defined in this study, of which, seven new species are established, i.e. Margattea deltodonta J-J He & Z-Q Wang, sp. nov., Margattea cuspidata J-J He & Z-Q Wang, sp. nov., Margattea caudata J-J He & Z-Q Wang, sp. nov., Margattea paratransversa J-J He & Z-Q Wang, sp. nov., Margattea disparilis J-J He & Z-Q Wang, sp. nov., Margattea transversa J-J He & Z-Q Wang, sp. nov., and Margattea bicruris J-J He & Z-Q Wang, sp. nov.


Membrane time constant as a tool to assess cell degeneration.
Changes in neuronal surface area may be monitored by measuring the plasma membrane capacitance [8]. Membrane time constant (tao m) is given by the product of the membrane resistance (rm) and membrane capacitance (Cm), tao m = rm Cm. Thus, when membrane resistance is kept constant at a steady state (resting), membrane time constant can reflect the size of neuronal surface area. Membrane time constant is the time for the potential to fall from the resting to a fraction (1-l/e), or 63%, of its final value in the charging curve during the application of a small negative current pulse. Negative voltage shift from the resting potential hardly activates any voltage-dependent ion channel, resulting in nominal changes in cell membrane resistance. Although elaborated methods for mathematical models and simulations are available for the electrophysiological assessment of neuron geometry in order to estimate subthreshold potential attenuation during the propagation of synaptically mediated electrical signals, they involve a number of critical assumptions for the convenience to each model, and some of these assumptions are unlikely to be valid. With these restrictive assumptions, very little can be determined about the electronic structure of a neuron beyond the measurement of neuronal membrane resistance and membrane time constant. Alternatively, numerous tracers are available to visualize morphologies of neurons intracellularly and extracellularly. These anatomical methods provide direct and quantitative evidence for neuron geometry; however, they involve tissue processing and a series of chemical reactions, some of which are time- and effort-demanding. The purpose of the present paper is to show that membrane time constant can be effectively used as a tool to assess diminution in cell surface area without involving extensive mathematical theories and/or neuroanatomical techniques. This approach is particularly effective in electrotonically compact cells such as hippocampal neurons. Recent development in the technique of the whole-cell patch clamp recording in the slice preparation yielded longer time constant with better resolution due to the absence of the leak conductance associated with microelectrode impalement. Indeed, when membrane time constant was measured with the whole-cell patch clamp recording technique, it successfully detected the reduction in dendritic arbors (dendritic degeneration) in dentate granule cells in the pilocarpine model of chronic epilepsy, and this finding is supported by the neuroanatomical evidence that was obtained from the same specimen samples. Membrane time constant is an easy-to-measure "passive membrane property" and can be used as a reliable probe by itself for detecting dendritic degeneration or as a tool for decision-making in introducing neuroanatomical technique in combination with slice neurophysiology.


Genetic and epigenetic stability of stem cells: Epigenetic modifiers modulate the fate of mesenchymal stem cells.
Stem cell research has progressed widely and has been receiving a considerable attention for its advantages and drawbacks. Despite their extensive therapeutic potential in regenerative medicine, they are debatable for their genetic and epigenetic stability. In fact lineage specific differentiation is mediated via epigenetic changes in DNA methylation, acetylation, histone modifications etc. Thus epigenetics plays an important role in stem cell biology. For therapeutic interventions stem cells need to be genetically and epigenetically stable for their maximum paracrine secretions for bringing about expected tissue repair and regeneration. In this review we have focused on the current status of genetic and epigenetic stability in stem cells and their importance in regenerative medicine. We have also touched upon the possibility of considering tissue resident mesenchymal stem cells as epigenetic modifiers. This is likely to open a new era in stem cell therapeutic intervention by reversing disease inducing epigenetic changes.


Progression of bone lesions in a child with primary hyperoxaluria type 1: evaluation by roentgenology and MRI.
We report the radiological and MRI findings of bone oxalosis and their rapid and extensive progression in a child with primary hyperoxaluria (PH) type 1. We emphasize the spectrum of skeletal changes and their differences from renal osteodystrophy and the role played by imaging in the diagnosis and follow-up of oxalosis.


Effect of 50% small bowel resection on gastric prostaglandin E2 levels in rats.
The mechanism for the production of increased gastric secretion following massive intestinal resection is not clearly defined. The loss of an intestinal inhibitor has been most frequently suggested to explain this hypersecretion. The role of endogenous prostaglandins which can inhibit gastric secretion is not established. The present study was undertaken to determine the effect of 50% proximal small bowel resection on Prostaglandin E2 (PGE2) levels in rat gastric mucosa. This study was performed in 30 rats divided into three groups. The first group of rats served as unoperated controls, the second group was sham operated and the third group underwent 50% resection of proximal small intestine. The PGE2 levels in rat gastric mucosa was decreased significantly (p < 0.001) in the resected group (422.85 +/- 7.66 pg/gm) as compared with the sham group (478.77 +/- 7.25 pg/gm) and the control group (493.38 +/- 4.61 pg/gm). Total gastric acidity was increased significantly (p < 0.001) in the resection group (63.05 +/- 2.64 mEq/L) as compared with the sham group (15.21 +/- 0.99 mEq/L) and the control group (17.19 +/- 0.80 mEq/L). The PGE2 levels and total gastric acidity were not significantly changed in either the control or sham operation groups (p > 0.05). The results suggest that endogenous prostaglandin synthesis has a regulatory role in gastric hyperacidity after 50% proximal small bowel resection in rats.


Methyltransferase-like 3 modulates osteogenic differentiation of adipose-derived stem cells in osteoporotic rats.
Osteoporosis (OP) is a metabolic bone disease involving reduced bone mass. Adipose-derived stem cells (ASCs) play an important role in bone regeneration. Emerging evidence suggests that methyltransferase-like 3 (METTL3) plays a significant role in bone development and metabolism. Therefore, this study investigates changes to METTL3 in the osteogenic differentiation of adipose stem cells in osteoporotic rats (OP-ASCs) and explores ways to enhance their osteogenic ability. An animal model of osteoporosis was established by removing both ovaries in rats. Real-time PCR and western blotting were performed to detect the expression of METTL3 and bone-related molecules, including runt-related transcription factor 2 (Runx2) and osteopontin (Opn). Furthermore, alkaline phosphatase staining was used to confirm the osteogenic potential of stem cells. Mettl3 small interfering RNA and Mettl3 overexpression lentivirus were used to assess the role of METTL3 in osteogenic differentiation of ASCs and OP-ASCs. The osteogenic differentiation capacity and Mettl3 expression significantly decreased in OP-ASCs. Moreover, Mettl3 silencing down-regulated the osteogenic ability of ASCs, and overexpression of Mettl3 recovered the impaired osteogenic capacity in OP-ASCs in vitro. The Mettl3 expression levels and osteogenic potential of OP-ASCs decreased. However, overexpression of METTL3 rescued the osteogenic ability of OP-ASCs, providing a new target for treatment of osteoporotic bone defects.


Sensitive analysis of fluorine and chlorine elements in water solution using laser-induced breakdown spectroscopy assisted with molecular synthesis.
Fluorine and chlorine are key elements to affecting water quality, but they are hard to be determined by conventional laser-induced breakdown spectroscopy (LIBS). To achieve high sensitivity detection of them, the CaF and CaCl molecules were synthesized by combining calcium in calcite and F and Cl in sample. The temporal characteristics of CaF and CaCl molecular emissions were investigated. It shows that molecular emission of CaF and CaCl has a longer lifetime and high spectral intensity than that of their atomic emissions. Such unique feature of molecular emission inspired us to use it for high sensitivity analysis of Cl and F elements in water. The results show that these two elements can be sensitively and accurately detected using LIBS assisted with molecular emission. The limits of detections (LoDs) were 0.38 mg/L and 1.03 mg/L for F and Cl elements, respectively, and the limit of quantitation (LoQ) was 3.404 mg/L to 20.569 mg/L for fluorine elements and 9.986 mg/L to 39.757 mg/L for fluorine. These detection limits can meet the World Health Organization's detection requirements for F and Cl elements in water. The results show that LIBS assisted with molecular synthesis has a huge potential in water quality monitoring.


Emergency ventilation using the Combitube in cases of difficult intubation.
The esophageal-tracheal Combitube (Sheridan, Argyle, NY) is a new device for emergency intubation, which can be inserted blindly without the use of a laryngoscope. Ventilation is independent of the position of the Combitube in either the esophagus or the trachea, since ventilation is always provided by the tube's double channel. The "tracheal" channel acts as a conventional endotracheal airway and has an open distal end. The "esophageal" channel has a blocked distal end, so that together with the inflated distal cuff it acts as an esophageal obturator in cases of esophageal intubation. Perforations at the pharyngeal section direct the airflow to the trachea. At the oropharyngeal section a large elastic balloon is positioned in order to obturate the oral cavity and the nasopharynx. Two patients are described to exemplify the Combitube's clinical use. Both had rapidly enlarging cervical hematomas causing upper airway obstruction and thus requiring immediate intubation. Endotracheal intubation failed because the glottis could not be visualized with a laryngoscope. In both cases the Combitube was applied successfully and adequate ventilation was provided via the Combitube placed esophageally. To better secure each patient's airway, tracheotomy was performed during ventilation without any complications.


Doxorubicin and CCNU with or without vincristine in patients with advanced refractory breast cancer. A randomized trial.
Thirty-five patients with advanced breast cancer refractory to initial chemotherapy were randomized to receive two-drug doxorubicin-CCNU or three-drug doxorubicin-CCNU-vincristine (VCR) treatment. Doxorubicin (25-40 mg/m2) and VCR (1.4 mg/m2) were given intravenously every 21 days; CCNU (65-90 mg/m2) was given orally every 42 days. Patients with liver or bone marrow metastases initially received the lower range doses. All patients failed prior chemotherapy with cyclophosphamide and 5-FU with or without methotrexate and prednisone; 67% received prior hormonal therapy. Pretreatment patient characteristics were comparable in both groups. Objective responses (greater than 50% reduction in tumor size) were seen in 7 of 18 patients (39%) on the VCR containing arm and in 8 of 17 patients (46%) on the doxorubicin-CCNU arm. Survival was not improved by VCR addition with overall survival on the doxorubicin-CCNU arm, approximately 10% greater at all intervals (median 9.1 versus 7.0 months; not statistically significant). However, neurotoxicity was significantly greater in the VCR arm (36% versus 0%; P less than 0.05). In advanced breast cancer, no benefit to VCR addition was seen in prior randomized studies of first-line combinations where VCR was the treatment variable. Such results, combined with our experience with VCR in a salvage regimen, question the value of VCR addition to combination regimens in advanced breast cancer.


m6A demethylase FTO induces NELL2 expression by inhibiting E2F1 m6A modification leading to metastasis of non-small cell lung cancer.
Non-small cell lung cancer (NSCLC) represents one of the primary causes of cancer-related mortality all over the world. Following our initial finding of the upregulated expression of E2F transcription factor-1 (E2F1) in the NSCLC-related microarray, this study aimed to explore the regulatory role of E2F1 and underlying mechanism in NSCLC development. NSCLC cell viability, migration, and invasion were evaluated utilizing Cell Counting Kit 8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), wound-healing, and Transwell assays. Loss- and gain-function assays were performed to determine the effects of the fat mass and obesity-associated protein (FTO)/E2F1/neural epidermal growth factor-like 2 (NELL2) axis on NSCLC cell behaviors in vitro and NSCLC tumor growth in vivo. E2F1 was highly expressed in both NSCLC tissues and cells. E2F1 augmented the viability, migration, and invasion of NSCLC cells, which was attributable to E2F1 transcriptionally activating NELL2. FTO upregulated the expression of E2F1 by inhibiting the m6A modification of E2F1. The FTO/E2F1/NELL2 axis modulated NSCLC cell viability, migration, and invasion in vitro as well as affected NSCLC tumor growth and metastasis in vivo. The FTO/E2F1/NELL2 axis may impart pro-tumorigenic effects on the cell behavior of NSCLC cells and thus accelerate NSCLC progression.


Sodium MRI of T1 High Signal Intensity in the Dentate Nucleus due to Gadolinium Deposition in Multiple Sclerosis.
Recently, several studies reported increased signal intensity (SI) of the dentate nucleus (DN) on unenhanced T1-weighted magnetic resonance imaging (MRI) as a possible consequence of multiple applications of gadolinium-based contrast agents. The aim of this study was to investigate with sodium (23 Na) MRI possible tissue abnormalities of the DN in multiple sclerosis (MS) patients. Sodium and conventional MRI were performed on a clinical 3T scanner. Total sodium concentrations (TSCs) of the DN, as well as DN-to-pons and DN-to-cerebellum SI ratios on unenhanced T1-weighted MRI were calculated. A total of 18 subjects (6/18 normal controls, 12/18 MS patients [6/12 with T1 hyperintense DN]) were investigated. There was no significant difference of TSC values of the DN in MS patients with a T1-hyperintense DN (33.70 ± 2.14 mM) compared to MS patients without those signal abnormalities (33.29 ± 1.67 mM; P = .67) or to healthy controls (33.14 ± 1.12; P = .32). Normal sodium signal in the T1-hyperintense DN in MS patients may point to relative tissue integrity despite gadolinium deposition in this area.


Identifying sensitive sources and key control handles for the reduction of greenhouse gas emissions from wastewater treatment.
This research investigates the effects of adjusting control handle values on greenhouse gas emissions from wastewater treatment, and reveals critical control handles and sensitive emission sources for control through the combined use of local and global sensitivity analysis methods. The direction of change in emissions, effluent quality and operational cost resulting from variation of control handles individually is determined using one-factor-at-a-time sensitivity analysis, and corresponding trade-offs are identified. The contribution of each control handle to variance in model outputs, taking into account the effects of interactions, is then explored using a variance-based sensitivity analysis method, i.e., Sobol's method, and significant second order interactions are discovered. This knowledge will assist future control strategy development and aid an efficient design and optimisation process, as it provides a better understanding of the effects of control handles on key performance indicators and identifies those for which dynamic control has the greatest potential benefits. Sources with the greatest variance in emissions, and therefore the greatest need to monitor, are also identified. It is found that variance in total emissions is predominantly due to changes in direct N2O emissions and selection of suitable values for wastage flow rate and aeration intensity in the final activated sludge reactor is of key importance. To improve effluent quality, costs and/or emissions, it is necessary to consider the effects of adjusting multiple control handles simultaneously and determine the optimum trade-off.


Use of methylene blue in vasoplegic syndrome that developed during non-cardiac surgery - A case report.
Vasoplegic syndrome is an increasingly recognized disease in perioperative medicine and is characterized by severe hypotension, normal or elevated cardiac output, and decreased systemic vascular resistance. It occurs commonly after cardiopulmonary bypass but may also occur after other types of surgery. Vasoplegic syndrome developed in our patient during posterior lumbar interbody fusion because of administering nicardipine after phenylephrine. However, the blood pressure did not increase as expected despite simultaneous use of norepinephrine and vasopressin to increase the reduced systemic vascular resistance. We present a case of vasoplegic syndrome that developed during posterior lumbar interbody fusion and was treated successfully with methylene blue.


RBM14 as a novel epigenetic-activated tumor oncogene is implicated in the reprogramming of glycolysis in lung cancer.
RNA-binding motif protein 14 (RBM14) is upregulated in a variety of tumors. However, the expression and biological role of RBM14 in lung cancer remain unclear. Chromatin immunoprecipitation and PCR were carried out to measure the levels of sedimentary YY1, EP300, H3K9ac, and H3K27ac in the RBM14 promoter. Co-immunoprecipitation was used to verify the interaction between YY1 and EP300. Glycolysis was investigated according to glucose consumption, lactate production, and the extracellular acidification rate (ECAR). RBM14 level is increased in lung adenocarcinoma (LUAD) cells. The increased RBM14 expression was correlated with TP53 mutation and individual cancer stages. A high level of RBM14 predicted a poorer overall survival of LUAD patients. The upregulated RBM14 in LUAD is induced by DNA methylation and histone acetylation. The transcription factor YY1 directly binds to EP300 and recruits EP300 to the promoter regions of RBM14, which further enhances H3K27 acetylation and promotes RBM14 expression. YY1-induced upregulation of RBM14 promoted cell growth and inhibited apoptosis by affecting the reprogramming of glycolysis. These results indicated that epigenetically activated RBM14 regulated growth and apoptosis by regulating the reprogramming of glycolysis and RBM14 may serve as a promising biomarker and therapeutic target for LUAD.


Epicutaneous testing.
The patch test and chamber test are the ordinary methods for detecting delayed type contact allergies. The method, test substances and criteria of positive reactions are already well standardized. Open testing is also used for problematic chemicals such as strong topical medicaments, emulsifiers and propylene glycol. The many problems of standardization are still unresolved in photocutaneous testing. Some chemicals are applied to the skin without occlusion, though the chamber test or patch test methods are used with most of them. The composition of the vehicle and occlusion time vary greatly from one photosensitizing substance to another. A xenon arc is recommended for screening any photosensitizers but fluorescent black light tubes are sufficient for routine clinical use. Type I contact allergies are far more common than was previously believed, especially among people allergic to birch pollen. This type of reaction and other forms of contact urticaria can be examined using the scratch chamber or scarification chamber techniques as well as with an open test on diseased or scratched (scarified) skin.


Sex differences and similarities in the neural circuit regulating song and other reproductive behaviors in songbirds.
In the 1970s, Nottebohm and Arnold reported marked male-biased sex differences in the volume of three song control nuclei in songbirds. Subsequently a series of studies on several songbird species suggested that there is a positive correlation between the degree to which there is a sex difference in the volume of these song control nuclei and in song behavior. This correlation has been questioned in recent years. Furthermore, it has become clear that the song circuit is fully integrated into a more comprehensive neural circuit that regulates multiple courtship and reproductive behaviors including song. Sex differences in songbirds should be evaluated in the context of the full complement of behaviors produced by both sexes in relation to reproduction and based on the entire circuit in order to understand the functional significance of variation between males and females in brain and behavior. Variation in brain and behavior exhibited among living songbird species provides an excellent opportunity to understand the functional significance of sex differences related to social behaviors.


Lipoprotein metabolism of pregnant women is associated with both their genetic polymorphisms and those of their newborn children.
To explore whether the placenta contributes to the lipoprotein metabolism of pregnant women, we took advantage of the fact that placental proteins are encoded from the fetal genome and examined the associations between lipids of 525 pregnant women and the presence, in their newborns, of genetic polymorphisms of LPL and apolipoprotein E (APOE), two genes expressed in placenta. After adjustment for maternal polymorphisms, newborn LPL*S447X was associated with lower triglycerides (-21 +/- 9 mg/dl), lower LDL-cholesterol (LDL-C; -12 +/- 5 mg/dl), lower apoB (-14 +/- 4 mg/dl), higher HDL-C (5 +/- 2 mg/dl), and higher apoA-I (9 +/- 4 mg/dl) in their mothers; newborn LPL*N291S was associated with higher maternal triglycerides (114 +/- 31 mg/dl); and newborn APOE*E2 (compared to E3E3) was associated with higher maternal LDL-C (14 +/- 6 mg/dl) and higher maternal apoB (14 +/- 5 mg/dl). These associations (all P < 0.05) were independent of polymorphisms carried by the mothers and of lipid concentrations in newborns and were similar in amplitude to the associations between maternal polymorphisms and maternal lipids. Such findings support the active role of placental LPL and APOE in the metabolism of maternal lipoproteins and suggest that fetal genes may modulate the risk for problems related to maternal dyslipidemia (preeclampsia, pancreatitis, and future cardiovascular disease).


Construction of a microfluidic platform integrating online protein fractionation, denaturation, digestion, and peptide enrichment.
Microfluidic system with multi-functional integration of high-throughput protein/peptide separation ability has great potential for improving the identification capacity of biological samples in proteomics. In this paper, a sample treatment platform was constructed by integrating reversed phase chromatography, immobilized enzyme reactor (IMER) and imprinted monolith through a microfluidic chip to achieve the online proteins fractionation, denaturation, digestion and peptides enrichment. We firstly synthesized a poly-allyl phenoxyacetate (AP) monolith and a lysine-glycine-glycine (KGG) imprinted monolith separately, and investigated in detail their performance in fractionating proteins and extracting KGG from the protein digests of MCF-7 cell. The removal percentage of 94.6% for MCF-7 cell protein and the recovery of 90.8% for KGG were obtained. The number of proteins and peptides identified on this microfluidic platform was 2,004 and 8,797, respectively, which was 2.8-fold and 3.0-fold higher than that of untreatment sample. The time consumed by this platform for a sample treatment was about 9.6 h, less than that of conventional method (approximate 13.3 h). In addition, this platform can enrich some peptide fragments containing KGG based on imprinted monolith, which can be served for the identification of ubiquitin-modified proteomics. The successful construction of this integrated microfluidic platform provides a considerable and efficient technical tool for simultaneous identification of proteomics and post-translational modification proteomics information.


[Changes of superoxide dismutase and lipid peroxide in lung tissue of rats after intratracheal injection of crocidolite and benzo(a) pyrene].
Wistar rats were divided into 4 groups: control group, Cro group, B(a) p group and Cro plus B(a) p group. Samples of lung tissue were collected 90, 180, 270, 360 and 540 days after the third time of intratracheal. The superoxide dismutase (SOD) activity, the level of lipid peroxide (LPO) and the ratio of SOD/LPO were observed. The results indicated that there was a synergistic action of Cro. and B(a) p.


Inhalation pharmacokinetics based on gas uptake studies. II. Pharmacokinetics of acetone in rats.
Inhalation pharmacokinetics of acetone in rats was studied using gas uptake techniques. Under conditions of negligible metabolism (saturation of metabolizing enzymes) the coefficient of distribution Keq between organism and gas phase was 220 which confirmed that acetone is mainly (but not exclusively) distributed within the body water compartment. The metabolic elimination of acetone from the animals followed strict Michaelis-Menten saturation kinetics; the km corresponded to 160 ppm in the atmosphere, Vmax was 230 mumol x h(-1) x kg(-1). A shift in the distribution pattern of acetone under conditions of prevalent metabolism was theoretically predicted and experimentally proven; as the partial process of metabolic elimination was dose-dependent, this "depleted" the animal compartment from acetone in a dose-dependent fashion causing a shift in the "steady-state-constant", Kst. Part of the acetone which was metabolically eliminated (4.7%) appeared in the urine as formate within 7 days after ending a 48-h exposure to acetone; this excretion was linear with time. The data suggested existence of a formate pool in the organism from which formate was released with delay. Hence, limitations may be inferred as to the applicability of urinary formate excretion as a quantitative indicator for changing conditions of acetone exposure.


Free-standing porous manganese dioxide/graphene composite films for high performance supercapacitors.
A simple hard template method and hydrothermal process have been employed to fabricate a self-standing hierarchical porous MnO2/graphene film. Thus-constructed electrode materials for binder-free supercapacitors exhibit a high specific capacitance of 266.3 F g(-1) at the density of 0.2 A g(-1). Moreover, the two-electrode device demonstrates an excellent rate capability and cycling stability with capacitance retention of 85.1% after 2000 charge-discharge cycles at a current density of 1 A g(-1). The porous nanostructured design can effectively improve the specific surface areas and account for the shorter relaxation time for the electrodes, resulting in a high electrochemical performance.


Oxygen pretreatment as protection against decompression sickness in rats: pressure and time necessary for hypothesized denucleation and renucleation.
Pretreatment with HBO at 300-500 kPa for 20 min reduced the incidence of decompression sickness (DCS) in a rat model. We investigated whether this procedure would be effective with lower oxygen pressures and shorter exposure, and tried to determine how long the pretreatment would remain effective. Rats were pretreated with oxygen at 101 or 203 kPa for 20 min and 304 kPa for 5 or 10 min. After pretreatment, the animals were exposed to air at 1,013 kPa for 33 min followed by fast decompression. Pretreatment at 101 or 203 kPa for 20 min and 304 kPa for 10 min significantly reduced the number of rats with DCS to 45%, compared with 65% in the control group. However, after pretreatment at 304 kPa for 5 min, 65% of rats suffered DCS. When pretreatment at 304 kPa for 20 min was followed by 2 h in normobaric air before compression and decompression, the outcome was worse, with 70-90% of the animals suffering DCS. This is probably due to the activation of "dormant" micronuclei. The risk of DCS remained lower (43%) when pretreatment with 100% O(2) at normobaric pressure for 20 min was followed by a 2 h interval in normobaric air (but not 6 or 24 h) before the hyperbaric exposure. The loss of effectiveness after a 6 or 24 h interval in normobaric air is related to micronuclei rejuvenation. Although pretreatment with hyperbaric O(2) may have an advantage over normobaric hyperoxia, decompression should not intervene between pretreatment and the dive.


Lewis basicity modulation of N-heterocycles: a key for successful cross-metathesis.
Cross-metathesis involving N-heteroaromatic olefinic derivatives is disclosed. The introduction of an appropriate substituent on the heteroaromatic ring decreases the Lewis basicity of the nitrogen atom, thus preventing the deactivation of the ruthenium-centered catalyst. The reaction is quite general in terms of both N-heterocycles and olefinic partners.


Storage stability studies on interesterified blend-based fast-frozen special fats for oxidative stability, crystallization characteristics and physical properties.
The storage stability of two kinds of interesterified blend-based fast-frozen special fats (PS:SO-IBSF, PS:RO-IBSF) with varied triacylglycerols (TAGs) compositions under different temperatures for 4 weeks was investigated. Rancimat and peroxide values experiments indicated that both IBSFs display good oxidation stability throughout a 4-week storage. As for the physical properties of both IBSFs, the solid fat content and hardness decreased with the increase of storage temperature, and IBSFs still exhibited a viscoelastic solid-like behavior. X-ray diffraction results showed that crystal transformation from β'- to β-form was more serious when stored at 25 °C. The more content of ECN 50-type TAGs in PS:RO-IBSF is helpful to reduce its crystal transformation from β'-to β-form compared to PS:SO-IBSF. On the other hand, storage at 4 °C was beneficial for both IBSFs to keep their crystal network integrity, and the PS:RO-IBSF maintained better quality under the same storage conditions.


Prognostic factors in idiopathic myelofibrosis: a simple scoring system with prognostic significance.
In a prognostic univariate analysis of a series of 80 patients with idiopathic myelofibrosis the Hb-concentration, the platelet count and osteomyelosclerosis emerged as factors with prognostic significance. A Hb-concentration less than 10 g/dl was associated with a significantly shorter survival than a Hb-concentration greater than or equal to 10 g/dl. A platelet count less than 100 x 10(9)/l also implied a significantly shorter survival. Patients with osteomyelosclerosis on X-ray of the skeleton had a significantly better prognosis as compared to those without osteomyelosclerosis. In a multivariate regression analysis the Hb-concentration consistently emerged as an important prognostic parameter, whereas the platelet count was only of prognostic significance within the first 6 months from diagnosis and the presence of osteomyelosclerosis emerged as a favourable parameter at 3 and 5 years. Based upon the above parameters and spleen size, a prognostic scoring system was designed which categorized the patients into three prognostic groups with highly different survival times (low risk group = 69 months; intermediate risk group = 33 months; high risk group = 4 months).


Monocyte chemoattractant protein-1-induced excitation and sensitization to mechanical stimulation of mechanosensitive C-fiber afferents in rat skin.
It has been previously demonstrated that chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2) increases the excitability of nociceptive neurons after peripheral nerve injury or inflammation. Moreover, decreased nocifensive mechanical threshold in behavioral tests and increased calcium influx in cultured dorsal root ganglion neurons by MCP-1 application have been reported. However, the effects of MCP-1 on peripheral afferent terminals have not been studied yet. The present study aimed to examine the effect of MCP-1 on the response of cutaneous unmyelinated afferents. For this purpose, single fiber recordings of mechanosensitive C-afferents were made in vitro from skin-saphenous nerve preparations excised from rats euthanized by CO2. Since IB4-positive neurons were previously implicated in MCP-1 induced mechanical hyperalgesia, sensitivity to α,β-methylene ATP (metATP), an indicator of IB4-positive neurons, was also studied. Application of MCP-1 100 ng/ml to the receptive field elicited excitation in one half of mechanosensitive unmyelinated afferents in the skin. MCP-1 also sensitized metATP insensitive fibers to mechanical stimulation, but not metATP sensitive fibers. The incidence of heat sensitive fibers was decreased in the MCP-1 treated group with a decrease in the response threshold. These results demonstrate MCP-1 is an effective stimulant of mechanosensitive unmyelinated peripheral afferents in the rat skin.


Chiral phosphoric acid catalyzed addition of dihydropyrans to N-acyl imines: stereocontrolled access to enantioenriched spirocyclic oxazoletetrahydropyrans with three contiguous stereocenters.
Dihydropyran derivatives readily undergo addition to N-acyl imines in the presence of chiral phosphoric acids. This addition process yields an attractive product that is capable of a tandem oxidative-cyclization via an epoxide intermediate.


Estimation of x-ray radiation related cancers in US dental offices: Is it worth the risk?
The objective of this study was to estimate the possible number of cancer cases produced during 2019 in US dental offices from radiography, estimate the possible reduction in those rates resulting from use of intraoral rectangular collimation and selection criteria, and determine the frequency and quality of website radiation risk information and informed consent forms. An analysis of dental radiation examinations in 2014 to 2015 US national survey data, Nationwide Evaluation of X-ray Trends, and National Council on Radiation Protection and Measurements surveys was performed, in addition to an analysis of 2008 to 2020 Journal of Clinical Orthodontics national orthodontic surveys for radiographic examination frequencies. Lifetime attributable cancer risk estimates from US and European studies were used to generate the total dental and orthodontic office cancer totals. In total, 150 offices were examined online for the quality and frequency of risk information in websites and consent forms. The 2019 estimate for all office cancers is 967. Collimation and selection criteria could reduce this to 237 cancer cases. Most cancers arise from intraoral and cone beam computed tomography examinations, with 135 orthodontic cancers over 21 months (average treatment time). Collimation and selection criteria could reduce this to 68. Only 1% of offices use collimators or informed consent for radiography. The website and consent information were of poor quality. Dentists are not following selection criteria or using collimators according to guidelines. Up to 75% of cancer cases could be avoided.


Pressure changes under the ischial tuberosities during gluteal neuromuscular stimulation in spinal cord injury: a comparison of sacral nerve root stimulation with surface functional electrical stimulation.
To compare the magnitude of interface pressure changes during gluteal maximus contraction by stimulating sacral nerve roots with surface electrical stimulations in patients with spinal cord injuries (SCIs). Pilot interventional study. Spinal injury research laboratory. Adults (N=18) with suprasacral complete SCI. Sacral nerve root stimulation (SNRS) via a functional magnetic stimulator (FMS) or a sacral anterior root stimulator (SARS) implant; and surface functional electrical stimulation (FES). Interface pressure under the ischial tuberosity (IT) defined as peak pressure, gradient at peak pressure, and average pressure. With optimal FMS, a 29% average reduction of IT peak pressure was achieved during FMS (mean ± SD: 160.1±24.3mmHg at rest vs 114.7±18.0mmHg during FMS, t5=6.3, P=.002). A 30% average reduction of peak pressure during stimulation via an SARS implant (143.2±31.7mmHg at rest vs 98.5±21.5mmHg during SARS, t5=4.4, P=.007) and a 22% average decrease of IT peak pressure during FES stimulation (153.7±34.8mmHg at rest vs 120.5±26.1mmHg during FES, t5=5.3, P=.003) were obtained. In 4 participants who completed both the FMS and FES studies, the percentage of peak pressure reduction with FMS was slightly greater than with FES (mean difference, 7.8%; 95% confidence interval, 1.6%-14.0; P=.04). SNRS or surface FES can induce sufficient gluteus maximus contraction and significantly reduce ischial pressure. SNRS via an SARS implant may be more convenient and efficient for frequently activating the gluteus maximus.


Association of maternal and umbilical cord blood leptin concentrations and abnormal color Doppler indices of umbilical artery with fetal growth restriction.
Fetal growth restriction (FGR) is a condition with heterogeneous pathophysiology which characterized by fetal weight less than the tenth percentile for gestational age. Several factors have impact on maternal, placental and fetal due to growth restriction. The aim of this study was to investigate the relationship between levels of leptin in the cord, and serum leptin of mothers also abnormal color Doppler indices of umbilical artery with fetal growth restriction. This is a cross sectional study conducted in Isfahan, Iran, 2015-2016. We recruited 40 women with singleton pregnancies complicated by fetal growth restriction (Group I) and 40 pregnant women with normal fetal growth (Group II) with matched age. Maternal serum and umbilical artery leptin levels were determined with Enzyme-Linked immunosorben method. Also, color Doppler ultrasound of umbilical artery was performed. Mean maternal and fetal leptin levels were lower in the FGR group compared to the normal group (36.58±(20.99) and 7.42 ±(4.08)vs. 47.32±(22.50) and 30.49±(14.50) respectively). Also, mean fetal leptin level was lower in the group with abnormal color Doppler sonographic indices compared to the normal group (7. 40 ±(4.10)vs 27.06±(15.80), respectively). This study indicated that maternal and fetal leptin levels are correlated with FGR originating from damaged placental function; also fetal leptin level can indicate changes in color Doppler sonographic indices.


Combining CUBIC Optical Clearing and Thy1-YFP-16 Mice to Observe Morphological Axon Changes During Wallerian Degeneration.
Wallerian degeneration is a pathological process closely related to peripheral nerve regeneration following injury, and includes the disintegration and phagocytosis of peripheral nervous system cells. Traditionally, morphological changes are observed by performing immunofluorescence staining after sectioning, which results in the loss of some histological information. The purpose of this study was to explore a new, nondestructive, and systematic method for observing axonal histological changes during Wallerian degeneration. Thirty male Thy1-YFP-16 mice (SPF grade, 6 weeks old, 20±5 g) were randomly selected and divided into clear, unobstructed brain imaging cocktails and computational analysis (CUBIC) optical clearing (n=15) and traditional method groups (n=15). Five mice in each group were sacrificed at 1st, 3rd, and 5th day following a crush operation. The histological axon changes were observed by CUBIC light optical clearing treatment, direct tissue section imaging, and HE staining. The results revealed that, compared with traditional imaging methods, there was no physical damage to the samples, which allowed for three-dimensional and deep-seated tissue imaging through CUBIC. Local image information could be nicely obtained by direct fluorescence imaging and HE staining, but it was difficult to obtain image information of the entire sample. At the same time, the image information obtained by fluorescence imaging and HE staining was partially lost. The combining of CUBIC and Thy1-YFP transgenic mice allowed for a clear and comprehensive observation of histological changes of axons in Wallerian degeneration.


Prognosis, trend and prediction in patient management.
A survey was conducted to get to know the 'state of the art' in trend prediction as a basis for optimal therapy, with emphasis on research being done in the countries of the European Community. Special areas of interest are quantitative prognosis (prognostic indices), the detection and use of trends in the patient's state, the use of models in prediction and their possible use for deciding which therapy is optimal for a specific patient. An extensive list of centres working in these fields as well as an extensive reference list is included.


Chitosan coating of zein-carboxymethylated short-chain amylose nanocomposites improves oral bioavailability of insulin in vitro and in vivo.
Non-invasive means of insulin administration circumvent some of the inconveniences of injections. Oral administration in particular is convenient, pain-free, and allows favorable glucose homeostasis, but is subject to chemical instability, enzymatic degradation, and poor gastrointestinal absorption. Natural polymeric nanoparticles have emerged as a promising oral delivery system for peptide therapeutics due their safety, biocompatibility, and stability. In this study, self-assembled nanocomposites from chitosan (CS) and insulin-loaded, zein-carboxymethylated short-chain amylose (IN-Z-CSA) nanocomposites were synthesized to improve oral bioavailability of insulin. The optimized IN-Z-CSA/CS0.2% nanocomposites exhibited an average size of 311.32±6.98 nm, a low polydispersity index (0.227±0.01), a negative zeta potential (43.77±1.36 mV), an encapsulation efficiency of 89.6±0.9%, and a loading capacity of 6.8±0.4%. The IN-Z-CSA/CS0.2% nanocomposites were stable in storage conditions. The transepithelial permeability of the N-Z-CSA/CS0.2% nanocomposites was 12-fold higher than that of insulin. Cellular uptake studies revealed that the IN-Z-CSA/CS0.2% nanocomposites were internalized into Caco-2 cells by both endocytosis and a paracellular route. Additionally, in pharmacological studies, orally administered IN-Z-CSA/CS0.2% nanocomposites had a stronger hypoglycemic effect with a relative bioavailability of 15.19% compared with that of IN-Z-CSA1.0% nanocomposites. Furthermore, cell toxicity and in vivo tests revealed that the IN-Z-CSA/CS0.2% nanocomposites were biocompatible. Overall, these results indicate that the IN-Z-CSA/CS0.2% nanocomposites can improve oral bioavailability of insulin and are a promising delivery system for insulin or other peptide/protein drugs.


Long-term delivery of protein therapeutics.
Proteins are effective biotherapeutics with applications in diverse ailments. Despite being specific and potent, their full clinical potential has not yet been realized. This can be attributed to short half-lives, complex structures, poor in vivo stability, low permeability, frequent parenteral administrations and poor adherence to treatment in chronic diseases. A sustained release system, providing controlled release of proteins, may overcome many of these limitations. This review focuses on recent development in approaches, especially polymer-based formulations, which can provide therapeutic levels of proteins over extended periods. Advances in particulate, gel-based formulations and novel approaches for extended protein delivery are discussed. Emphasis is placed on dosage form, method of preparation, mechanism of release and stability of biotherapeutics. Substantial advancements have been made in the field of extended protein delivery via various polymer-based formulations over last decade despite the unique delivery-related challenges posed by protein biologics. A number of injectable sustained-release formulations have reached market. However, therapeutic application of proteins is still hampered by delivery-related issues. A large number of protein molecules are under clinical trials, and hence, there is an urgent need to develop new methods to deliver these highly potent biologics.


Isolation of human T and B lymphocytes by rosette formation with 2-aminoethylisothiquronium bromide (AET) -treated sheep red blood cells with monkey red blood cells.
Purified human B and T lymphocytes were obtained by rosetting HPL with AET-SRBC or MRBC and separating the non-rosetted from the rosetted cells on Ficoll-Hypaque gradient. 92 +/- 5% of the B cells were fluorescent positive for mBlg and 95 +/- 2% of the purified T cells rosetted with AET-SRBC. 69 +/- 5% of the B cells and 61 +/- 8% of the T cells present in the unifractionated HPL were recovered in the purified fractions.


Exosomes from M1-Polarized Macrophages Potentiate the Cancer Vaccine by Creating a Pro-inflammatory Microenvironment in the Lymph Node.
Exosomes are small membrane-bound vesicular particles generated by most cells for intercellular communication and regulation. During biogenesis, specific lipids, RNAs, proteins, and carbohydrates are enriched and packaged into the vesicles so that the exosomal contents reflect not only the source but also the physiological conditions of the parental cells. These exosomes transport materials or signals to the target cells for diverse physiological purposes. Our study focused on the exosomes derived from M1-polarized, proinflammatory macrophages for the possibility of using M1 exosomes as an immunopotentiator for a cancer vaccine. The M1 exosomes displayed a tropism toward lymph nodes after subcutaneous injection, primarily taken up by the local macrophages and dendritic cells, and they induced the release of a pool of Th1 cytokines. We found that M1, but not M2, exosomes enhanced activity of lipid calcium phosphate (LCP) nanoparticle-encapsulated Trp2 vaccine, and they induced a stronger antigen-specific cytotoxic T cell response. The M1 exosomes proved to be a more potent immunopotentiator than CpG oligonucleotide when used with LCP nanoparticle vaccine in a melanoma growth inhibition study. Thus, our study indicated that exosomes derived from M1-polarized macrophages could be used as a vaccine adjuvant.


Channelrhodopsins-Their potential in gene therapy for neurological disorders.
Recently, channelrhodopsins (ChRs) have begun to be used to manipulate the neuronal activity, since they can be targeted to specific neurons or neural circuits using genetic methods. To advance the potential applications in the investigation and treatment of neurological disorders, the following types of basic research should receive extensive financial support. The spectral and kinetic properties of ChRs should be optimized according to the application by generating variants of ChRs or exploring new rhodopsins from other species. These ChRs should be targeted to the specific types of neurons involved in the neurological disorders through a gene expression system using cell- or tissue-specific promoters/enhancers as well as gene delivery systems with modified virus vectors. The methods have to be developed to apply the genes of interest with safety and long-term effectiveness. Sophisticated opto-electrical devices should be developed. Appropriate primate animal model systems should be established to minimize the structural differences between small animals such as rodents and human beings. In this paper, we will review the current progress in the basic research concerned with the potential clinical application of ChRs and discuss the future directions of research on ChRs so that they could be applied for human welfare.


Phlegmasia Cerulea Dolens in the Upper Extremity: A Case Report and Literature Review.
Phlegmasia cerulea dolens (PCD) is a rare condition characterized by deep venous thrombosis with gangrene. It can result in critically severe edema that affects the blood supply to the limbs. PCD generally occurs in the lower rather than upper extremity. We herein present a case report of upper extremity PCD and discuss thrombophilia secondary to low protein S activity as the main cause. Catheter-directed thrombolysis via the occluded end of the artery may be one of the best treatment methods for PCD.


Effect of lactic acid on L-glutamate uptake in cultured astrocytes: mechanistic considerations.
Elevated levels of lactic acid can be deleterious to CNS tissue. Lactic acid is known to cause astroglial swelling and since glial swelling has been shown to inhibit L-glutamate (L-Glu) uptake, we examined whether one of the actions of lactic acid is to inhibit L-Glu uptake. Astrocyte cultures treated with lactic acid (25 mM; pH 6.1) showed an inhibition of L-Glu uptake by 65%. HCl (pH 6.1) also inhibited L-Glu uptake and this inhibition was potentiated by sodium lactate (25 mM). The inhibitory effect of lactic acid on L-Glu uptake was partially reversible and the reversibility was enhanced by hypothermia. Blocking glial swelling with D-mannitol, or treatment with antioxidants or hypothermia did not inhibit the effect of lactic acid on L-Glu uptake, indicating that swelling per se or free radicals, were not the factors in L-Glu uptake inhibition. Lactic acid induced a four-fold enhancement of L-Glu release and a seven-fold increase of K+ release. Our results suggest that lactic acid, by direct effect on pH, brings about a stimulation of K+ and L-Glu release which may be a factor in the inhibition of L-Glu uptake by lactic acid in astrocytes.


Anterior cingulate activation and error processing during interferon-alpha treatment.
There has been increasing interest in the role of immunologic processes, notably cytokines, in the development of behavioral alterations, especially in medically ill patients. Interferon (IFN)-alpha is notorious for causing behavioral symptoms, including depression, fatigue, and cognitive dysfunction, and has been used to investigate the effects of cytokines on the brain. In the present study we assessed the effects of low-dose IFN-alpha on brain activity, using functional magnetic resonance imaging during a task of visuospatial attention in patients infected with hepatitis C virus (HCV). Despite endorsing symptoms of impaired concentration and fatigue, IFN-alpha-treated patients (n = 10) exhibited task performance and activation of parietal and occipital brain regions similar to that seen in HCV-infected control subjects (n = 11). Interestingly, however, in contrast to control subjects, IFN-alpha-treated patients exhibited significant activation in the dorsal part of the anterior cingulate cortex (ACC), which highly correlated with the number of task-related errors. No such correlation was found in control subjects. Consistent with the role of the ACC in conflict monitoring, ACC activation during IFN-alpha administration suggests that cytokines might increase processing conflict or reduce the threshold for conflict detection, thereby signaling the need to exert greater mental effort to maintain performance. Such alterations in ACC activity might in turn contribute to cytokine-induced behavioral changes.


Temporal context and conditional associative learning.
We investigated how temporal context affects the learning of arbitrary visuo-motor associations. Human observers viewed highly distinguishable, fractal objects and learned to choose for each object the one motor response (of four) that was rewarded. Some objects were consistently preceded by specific other objects, while other objects lacked this task-irrelevant but predictive context. The results of five experiments showed that predictive context consistently and significantly accelerated associative learning. A simple model of reinforcement learning, in which three successive objects informed response selection, reproduced our behavioral results. Our results imply that not just the representation of a current event, but also the representations of past events, are reinforced during conditional associative learning. In addition, these findings are broadly consistent with the prediction of attractor network models of associative learning and their prophecy of a persistent representation of past objects.


A systematic review and meta-analysis of trials assessing PD-1/PD-L1 immune checkpoint inhibitors activity in pre-treated advanced stage malignant mesothelioma.
Advanced stage malignant mesothelioma (asMM) patients have poor prognosis. Several trials investigated the role of programmed cell death protein-1 (PD-1) and its ligand 1 (PD-L1) immune checkpoint inhibitors (ICIs) in pre-treated asMM. A systematic review of the literature of clinical trials testing single-agent anti PD-1/PD-L1 ICIs in pre-treated asMM was performed. Objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS) and overall survival (OS) data were extracted. The predictive role of PD-L1 was assessed. We selected 13 studies including 888 patients. ORR and DCR were 18.1% (95% confidence interval [CI] 13.9-22.8%) and 55.4% (95% CI: 48.1-62.5%), respectively. Median PFS and OS ranged from 2.1 to 5.9 and from 6.7 to 20.9 months, respectively. ORR according to PD-L1 was 27.0% (95% CI: 18.7-36.2%). Anti-PD-(L)1 ICIs might be considered a treatment option for chemotherapy-resistant asMM, even if reliable predictive factors are still lacking.


Effect of the Piezoelectric Device on Intraoperative Hemorrhage Control and Quality of Life after Endodontic Microsurgery: A Randomized Clinical Study.
The purpose of this study was to evaluate the effect of the piezoelectric device on intraoperative hemorrhage control during surgery and the quality of life of patients after endodontic microsurgery. A total of 40 patients were randomly divided into the piezo group (n = 20) and the control group (n = 20). In the piezo group, after flap reflection, bone cutting, granulation tissue removal, and root-end resection were performed using the piezoelectric surgical device and surgical carbide burs, and curettes were used in the control group. The quality of life of patients was evaluated daily for 1 week postsurgery for limitations of oral and general functions, pain, and other symptoms. Limitation of functions and other symptoms were recorded by a modified version of the patient's perception questionnaire using a 5-point Likert scale for mouth opening, chewing, speaking, sleeping, daily routine, missed work, swelling, nausea, and bad taste/breath, and the visual analog scale was adopted for pain. Hemorrhage control during surgery was independently assessed by the surgeon and 2 blinded observers and recorded as 0 (no hemorrhage control), 1 (intermittent control), and 2 (complete control). The chi-square test was used to assess hemorrhage control. For variables related to function and symptoms other than pain and analgesics taken, the Fisher exact test was used. For the assessment of pain between the 2 groups, the Mann-Whitney U test was used. For parameters of quality of life, the piezo group showed significantly less swelling on the first, second, and third days and pain on the first and second days compared with the control group (P < .05). Analgesics taken were also significantly less in the piezo group (P < .05). In the piezo group, complete hemorrhage control was achieved in 10 patients, and in the control group, it was achieved only in 1 patient (P < .05). Piezoelectric surgery resulted in improved quality of life of patients in the first week postsurgery with lower levels of pain and swelling as well as the number of analgesics taken and better hemorrhage control during surgery.


Surface stiffness depended gingival mesenchymal stem cell sensitivity to oxidative stress.
Mesenchymal stem cells (MSCs) are widely used in the fields of cell therapy and tissue engineering, due to their wide spectrum of differentiation potential, immunomodulation function and ongoing oxidative stress (OS) reduction. Nevertheless, OS impact is often overlooked in these research fields. It is not only responsible for the induction and development of many ailments, e.g., diabetes, lung fibrosis, and cancer, moreover, OS causes stem cell death and senescence during cell therapy and tissue engineering practices. As MSCs are used to treat various tissues, they interact with different tissue-specific mechanical environments, thus it is important to understand how the mechanical environment impacts MSC sensitivity to OS. In this work, for the first time, as known to the authors, it was shown that gingival MSCs (GMSCs) sensitivity to OS depends on the stiffness of the surface, on which the cells are grown. Furthermore, the activity and expression of mitogen activated protein kinases ERK, JNK, and p38 were surface stiffness dependent. GMSCs isolated from intermediate/stiff gingiva tissue (~20 kPa) have shown the best proliferative and survival properties, then grown on the stiffest tissues mimicking polyacrylamide hydrogels (40 kPa). Therefore, MSC source might determine their sensitivity to OS in different stiffness environments and should be accounted when developing a treatment strategy.


Microbial transformation of pharmaceuticals naproxen, bisoprolol, and diclofenac in aerobic and anaerobic environments.
Although biotransformation is generally considered to be the main process by which to remove pharmaceuticals, both in sewage treatment plants and in aquatic environments, quantitative information on specific compounds is scarce. In this study, the transformations of diclofenac (DCF), naproxen (NPX), and bisoprolol (BSP) were studied under aerobic and anaerobic conditions using inocula taken from activated and digested sludge processes, respectively. Whereas concentration decays were monitored by LC-tandem mass spectrometry, oxygen consumption and methane production were used for the evaluation of the performance of overall conditions. DCF was recalcitrant against both aerobic and anaerobic biotransformation. More than one third of the BSP disappeared under aerobic conditions, whereas only 14% was anaerobically biotransformed in 161 days. Under aerobic conditions, complete removal of NPX was evident within 14 days, but anaerobic transformation was also efficient. Formation of 6-O-desmethylnaproxen, a previously reported aerobic metabolite, was also detected under anaerobic conditions and persisted for 161 days.


Varying the Directionality of Protein Catalysts for Aldol and Retro-Aldol Reactions.
Natural aldolase enzymes and created retro-aldolase protein catalysts often catalyze both aldol and retro-aldol reactions depending on the concentrations of the reactants and the products. Here, we report that the directionality of protein catalysts can be altered by replacing one amino acid. The protein catalyst derived from a scaffold of a previously reported retro-aldolase catalyst, catalyzed aldol reactions more efficiently than the previously reported retro-aldolase catalyst. The retro-aldolase catalyst efficiently catalyzed the retro-aldol reaction but was less efficient in catalyzing the aldol reaction. The results indicate that protein catalysts with varying levels of directionality in usually reversibly catalyzed aldol and retro-aldol reactions can be generated from the same protein scaffold.


Comparing intraoperative parathyroid identification based on surgeon experience versus near infrared autofluorescence detection - A surgeon-blinded multi-centric study.
Near infrared autofluorescence (NIRAF) detection has previously demonstrated significant potential for real-time parathyroid gland identification. However, the performance of a NIRAF detection device - PTeye® - remains to be evaluated relative to a surgeon's own ability to identify parathyroid glands. Patients eligible for thyroidectomy and/or parathyroidectomy were enrolled under 6 endocrine surgeons at 3 high-volume institutions. Participating surgeons were categorized based on years of experience. All surgeons were blinded to output of PTeye® when identifying tissues. The surgeon's performance for parathyroid discrimination was then compared with PTeye®. Histology served as gold standard for excised specimens, while expert surgeon's opinion was used to validate in-situ tissues. PTeye® achieved 92.7% accuracy across 167 patients recruited. Junior surgeons (<5 years of experience) were found to have lower confidence in parathyroid identification and higher tissue misclassification rate per specimen when compared to PTeye® and senior surgeons (>10 years of experience). NIRAF detection with PTeye® can be a valuable intraoperative adjunct technology to aid in parathyroid identification for surgeons.


Anatomical and functional outcomes after bilateral sacrospinous colposuspension (BSC) for the treatment of female genital prolapse.
Pelvic organ prolapse is a bothersome condition affecting many women at advanced age, but also frequently observed in young women with certain risk factors. Various surgical techniques have been developed with the aim of providing effective surgical treatment for apical prolapse. The vaginal bilateral sacrospinous colposuspension surgery (BSC) with ultralight mesh and utilization of the i- stich is a relatively new minimal invasive technique with very promising outcomes. The technique offers apical suspension, in the presence or absence of the uterus. The objective of this study is to evaluate the anatomical and functional outcomes of bilateral sacrospinous colposuspension with ultralight mesh in 30 Patients treated with the vaginal single incision standardized technique. In this retrospective study, 30 patients were treated by BSC for significant vaginal, uterovaginal or cervical prolapse. A simultaneous anterior colporrhaphy, posterior colporrhaphy or both were performed when indicated. Anatomical and functional outcomes were assessed 1 year postoperatively using the Pelvic Organ Prolapse Quantification system (POP-Q) and the standardised Prolapse Quality of Life (P-QOL) questionnair. The POP-Q parameters were significantly improved at twelve months after surgery compared to baseline. The total score and all four subdomains of the P-QOL-questionnaire showed positive trends and improvement at twelve months after surgery when compared to preoperative values. All patients were asymptomatic and expressed high satisfaction one year after surgery. No intraoperative adverse events were recorded for all patients. Only minimal postoperative complications were recorded and they all resolved completely with conservative management. This study highlights the functional and anatomical outcomes of the minimally invasive vaginal bilateral sacrospinal colposuspension with ultralight mesh for the management of apical prolapse. The one year postoperative results of the proposed procedure reflect excellent outcomes with minimal complications. The data published here are very promising and warrant further investigations and more studies to evaluate the long-term outcomes of BSC in the surgical management of apical defects. The study protocol was approved by the Ethics Committee at the University Hospital of Cologne, Germany (Date of registration: 08.02.2022) (Registration number: 21-1494-retro) (retrospectively registered).


A method for administration of prolonged intravenous infusion of prostacyclin (PGI2) to unanesthetized rats.
Prostacyclin is short acting and chemically unstable. To study sustained effects in an intact animal, prolonged intravenous infusion may be required. The compound has adequate stability for 24 hr in pH 10.0 carbonate buffer at 0 degrees. A "displacement syringe" is described wherein the prostacyclin solution is stored in a rubber bag inside the barrel of a 5 ml syringe. This device is placed in an ice bath. A syringe pump drives water into the barrel displacing an equal volume of solution out of the bag. Chronic venous cannules, saddles, and flow-through swivels are used as for drug self-administration studies. A simple, inexpensive rack for use with conventional individual hanging cages is also described.


Three-dimensional photon treatment planning of the intact breast.
Three-dimensional treatment planning for the intact breast was performed on two patients who had undergone CT scanning. A total of 38 treatment plans were evaluated. Multiple plans were evaluated for each patient including plans with and without inhomogeneity corrections, plans using varying photon energies of 60Co, 4 MV, 6 MV, 10 MV, and 15 MV, and three-dimensionally unconstrained plans. Increased hot spots were appreciated in the central axis plane when lung inhomogeneity corrections were used. Additional hot spots were appreciated in off-axis planes towards the cephalad and caudad aspects of the target volume because of lung inhomogeneity corrections and changes in the breast contour. The use of 60Co was associated with an increase in the magnitude and volume of hot spots, whereas the use of higher energy photons such as 10 MV and 15 MV was associated with an unacceptable target coverage at shallow depths. Therefore, for the two patients studied, the use of a medium energy photon beam (such as from a 6 MV linear accelerator) appeared to be the energy of choice for treatment of the intact breast. The three-dimensionally unconstrained plans were able to improve slightly upon the standard plans, particularly with relationship of dose to normal tissue structures. Areas for future research were identified, including the use of tissue compensators.


Evaluating a Brief, Video-Based Sexual Risk Reduction Intervention and Assessment Reactivity with STI Clinic Patients: Results from a Randomized Controlled Trial.
We report results from a randomized controlled trial designed to evaluate the efficacy of a video-based sexual risk reduction intervention and to measure assessment reactivity. Patients (N = 1010; 56 % male; 69 % African American) receiving care at a sexually transmitted infection (STI) clinic were assigned to one of four conditions formed by crossing assessment condition (i.e., sexual health vs. general health) with intervention condition (i.e., sexual risk reduction intervention vs. general health promotion). After completing their assigned baseline assessment, participants received their assigned intervention, and subsequently returned for follow-up assessments at 3, 6, 9, and 12 months. Participants in all conditions reduced their self-reported sexual risk behavior, and the incidence of new STIs declined from baseline through the follow-ups; however, there was no effect of intervention or assessment condition. We conclude that further risk reduction will require more intensive interventions, especially in STI clinics that already provide excellent clinical care.


Novel integrated 3D multidetector computed tomography and fluoroscopy fusion for left atrial appendage occlusion procedures.
This report demonstrates the application and feasibility of novel 3D-MDCT real-time fusion technology with fluoroscopy, for left atrial appendage (LAA) occlusion procedures. A successful LAA occlusion procedure relies on multiple imaging modalities, including TEE or 3D-MDCT, and fluoroscopy. Effectively integrating these imaging modalities may improve implantation safety and success. To our knowledge this technique has not been previously described for LAA occlusions. This observational study compared clinical and procedural parameters for procedures performed with or without fusion integration. All patients had a pre-procedural 3D-MDCT for LAA measurements, along with 3D analyses of LAA morphology and surrounding structures. Using the image fusion software (Valve ASSIST 2, GE Healthcare, UK), landmarks were identified on fluoroscopy, and MDCT LAA anatomy outlines were then projected onto the real-time fluoroscopy image during the procedure, to guide all steps of the intervention. A total of 57 patients underwent LAA occlusion, with 16 performed using fusion software. In comparison to the pre-fusion group, reductions in contrast volume (21.0 ± 11.7 vs. 95.9 ± 80.5 ml, P < 0.001), procedure time (63.0 ± 22.0 vs. 87.3 ± 43.0 min, P = 0.01), and fluoroscopy time (6.2 vs. 8.3 min, P = 0.03) were observed. Incomplete sealing (0 vs. 14.6%, P = 0.16) and device deployment success (100 vs. 92.7%, P = 0.17) were not significantly different. The addition of this novel fusion technology is safe and feasible. To optimize LAA procedural success, fusion integration may offer a promising addition, or alternative, to current imaging modalities. © 2017 Wiley Periodicals, Inc.


Sequencing errors in Native American mitogenomes: Impact on clade definitions, haplogroup assignation, and beyond.
Available evidence allows the interpretation that some cases of absence of otherwise expected variation, based on phylogenetic expectations in mitogenomes of Native American origin, are due to artificial recombination rather than to homoplasy, while other more complex scenarios involving combination of original Cambridge Reference Sequence mistakes plus incomplete or incorrect scoring of variation are also showed. Several instances of mismatched control and coding regions as well as partially duplicated HV2 are observed in Peruvians, while intra-haplogroup chimaeras of different D1 subhaplogroups are referred to in Mexican Native Americans. A revised definition for haplogroup B2h is proposed, and preventive quality control measures are suggested.


In vitro discovery of novel prokaryotic ion channel candidates for antiarrhythmic gene therapy.
Sudden cardiac death continues to have a devastating impact on public health prompting the continued efforts to develop more effective therapies for cardiac arrhythmias. Among different approaches to normalize function of ion channels and prevent arrhythmogenic remodeling of tissue substrate, cardiac cell and gene therapies are emerging as promising strategies to restore and maintain normal heart rhythm. Specifically, the ability to genetically enhance electrical excitability of diseased hearts through voltage-gated sodium channel (VGSC) gene transfer could improve velocity of action potential conduction and act to stop reentrant circuits underlying sustained arrhythmias. For this purpose, prokaryotic VGSC genes are promising therapeutic candidates due to their small size (<1kb) and potential to be effectively packaged in adeno-associated viral (AAV) vectors and delivered to cardiomyocytes for stable, long-term expression. This article describes a versatile method to discover and characterize novel prokaryotic ion channels for use in gene and cell therapies for heart disease including cardiac arrhythmias. Detailed protocols are provided for: (1) identification of potential ion channel candidates from large genomic databases, (2) candidate screening and characterization using site-directed mutagenesis and engineered human excitable cell system and, (3) candidate validation using electrophysiological techniques and an in vitro model of impaired cardiac impulse conduction.


Changes in interstitial collagen metabolism during acute myocardial infarction treated with streptokinase or tissue plasminogen activator.
Collagen plays a specific role in the maintenance of vascular integrity and in the thrombosis and scar formation processes. Therefore we found it interesting to study the changes in interstitial collagen metabolism during acute myocardial infarction treated with thrombolytic agents. Changes in collagen synthesis were evaluated by obtaining assays of the serum concentrations of the carboxyterminal propeptide of type I procollagen. Except fibrin plasmin is capable of degrading extracellular matrix components including collagen, and this capability was evaluated by monitoring the serum concentrations of the aminoterminal propeptide of type III procollagen. Twenty-four patients with suspected acute myocardial infarction and indications for thrombolytic therapy were randomized to receive either streptokinase (n = 11) or tissue plasminogen activator (n = 13). The patient groups were identical in their clinical characteristics. Serum levels of the aminoterminal propeptide of type III collagen increased rapidly on infusion of the thrombolytic agents, with the maximal mean increases of 44% and 16% in the streptokinase and TPA-treated groups, respectively. Levels of the carboxyterminal propeptide of type I collagen did not change during the thrombolytic therapy. A transient decrease occurred in the type I propeptide concentration at postinfarction day 2, and this decrease was followed by a secondary increase at days 4 to 6 in both patient groups studied. We conclude that thrombolytic agents stimulate the breakdown of interstitial collagen and that the collagen-degrading activity of TPA is lower than that of streptokinase. This factor may contribute to the relatively higher rethrombosis rate seen after TPA, because exposed collagen in the affected vascular wall stimulates thrombosis formation. On the other hand, increased collagen degradation followed by inhibition of collagen synthesis in the infarcted myocardium might increase the risk for cardiac rupture, especially after streptokinase treatment.


Propagation of mouse mammary tumor cell lines and production of mouse mammary tumor virus in a serum-free medium.
Five different mouse mammary tumor cell lines were propagated in a serum free medium. Evaluation of growth characteristics, including logarithmic growth, cell population increase, protein production and days to confluency, showed serum-free medium comparable to serum-containing medium. Mouse mammary tumor virus expression and production, in C3H and GR tumor cell lines, as determined by virus particle counting and RNA dependent DNA polymerase assays, subsequent to dexamethasone stimulation revealed equivalent to higher levels of virus in serum-free medium as compared to serum-containing medium.


Genetics of Food Allergy.
The risk factors for food allergy (FA) include both genetic variants and environmental factors. Advances using both candidate-gene association studies and genome-wide approaches have led to the identification of FA-associated genes involved in immune responses and skin barrier functions. Epigenetic changes have also been associated with the risk of FA. In this chapter, we outline current understanding of the genetics, epigenetics and the interplay with environmental risk factors associated with FA. Future studies of gene-environment interactions, gene-gene interactions, and multi-omics integration may help shed light on the mechanisms of FA, and lead to improved diagnostic and treatment strategies.


Optimal schemes of radial network arch pedestrian bridges: An extensive dataset of solutions under different conditions.
This data article provides a series of 220 optimal parameter sets of steel radial network arch pedestrian bridge schemes. Each set includes 10 design parameters for a two-dimensional bridge frame: the arch rise, the number of hangers, their (variable) spread and central angles, and the dimensions of the arch, girder, and hangers. Additionally, solutions are provided for different initial conditions such as constant hanger angles, given ratio values between the arch rise and the bridge span, given hanger diameters, and for the condition of hangers' verticality. These data are related to the research article "Parametric study on mass minimization of radial network arch pedestrian bridges" (Belevičius et al., 2021). In this paper, the bridge scheme was optimized seeking for the minimal bridge mass under different loading cases according to recommendations of Eurocode EN 1991-2. Since the optimization problem is multimodal, and the employed stochastic global optimization algorithms provide different solutions for each run of the algorithm, we render the five best parameters' sets for every problem. In many cases, close objective function values correspond to fairly distinct parameter values. This dataset could be used by bridge designers as an initial design hint by choosing the appropriate parameters set.


Separation of the potential G-quadruplex ligands from the butanol extract of Zanthoxylum ailanthoides Sieb. & Zucc. by countercurrent chromatography and preparative high performance liquid chromatography.
G-quadruplex DNA structure is considered to be a very attractive target for antitumor drug design due to its unique role in maintaining telomerase activities. Therefore, discovering ligands with high stability of G-quadruplex structure is of great interest. In this paper, pH-zone refining counter current chromatography (CCC) and preparative high performance liquid chromatography (HPLC) were employed for the separation of potent G-quadruplex ligands from the n-butanol fraction of the crude extract of Zanthoxylum ailanthoides, which is a traditional Chinese medicine recently found to display high inhibitory activity against several human cancer cells. The 75% aqueous ethanol extract of the stem bark of Z. ailanthoides and its fractions with petroleum ether, ethyl acetate and n-butanol displayed almost the same G-quadruplex stabilization ability. Here, pH-zone refining CCC was used for the separation of the alkaloids from the n-butanol fraction by a seldom used solvent system composed of dichloromethane-methanol-water (4:1:2.5) with 10mM TEA in the organic stationary phase as retainer and 10mM HCl in the aqueous mobile phase as eluter. Compounds I, II and III were obtained, with purity greater than 95%, in the quantities of 31.2, 94.0, and 26.4mg respectively from 300mg of lipophilic fraction within 80min, which were identified as three tetrahydroprotoberberines isolated for the first time in this plant. In addition, a phenylpropanoid glycoside compound IV (Syringin), an isoquinoline (Magnoflorine, V), and two lignin isomers (+)-lyoniresiol-3α-O-β-d-glucopyranoside (VI) and (-)-lyoniresinol -3α-O-β-D -glucopyranoside (VII) were isolated by traditional CCC together with preparative HPLC. Compounds IV, V, VI and VII were obtained, with purity greater than 95%, in the quantities of 4.0, 13.2, 6.7, and 6.5mg respectively from 960mg of hydrophilic fraction. Among the seven isolated compounds, tetrahydroprotoberberine I, II and III were found to display remarkable stabilization effects on G-quadruplex by increasing G-quadruplex's Tm approximately 10°C, which may be the most potent G-quadruplex ligands in Z. ailanthoides.