The effect of tetracyclines on quantitative measures of osteoclast morphology.
We report the effects of the tetracycline analogues 4-dedimethylaminotetracycline (CMT-1) and minocycline on osteoclast spreading and motility. Both agents influenced the morphometric descriptor of cell spread area, rho, producing cellular retraction or an R effect (half-times: 30 and 44 minutes for CMT-1 and minocycline, respectively). At the concentrations employed, the tetracycline-induced R effects were significantly slower than, but were qualitatively similar to, those resulting from Ca2+ "receptor" activation through the application of 15 mM-[Ca2+] (slopes: -1.25, -0.18, and -4.40/minute for 10 mg/l-[CMT-1], 10 mg/l-[minocycline] and 15 mM-[Ca2+], respectively). In contrast, the same tetracycline concentrations did not influence osteoclast margin ruffling activity as described by mu, a motility descriptor known to be influenced by elevations of cellular cyclic AMP. Thus, the tetracyclines exert morphometric effects comparable to changes selectively activated by occupancy of the osteoclast Ca2+ "receptor" which may act through an increase in cytosolic [Ca2+].


Adenine-coated magnetic multiwalled carbon nanotubes for the selective extraction of aristolochic acids based on multiple interactions.
A method is described for the functionalization of magnetic carbon nanotubes to recognize aristolochic acid Ⅰ and Ⅱ. 3-Glycidyloxypropyltrimethoxysilane was used as a coupling agent to immobilize adenine on a solid support. The morphology and structure of adenine-coated magnetic carbon nanotubes was investigated using transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD) and a vibrating sample magnetometer (VSM). The adsorption performance of the adenine-coated magnetic carbon nanotubes was evaluated via adsorption isotherms, the kinetics and selectivity tests. The adsorption capacity of the adenine-functionalized sorbent for aristolochic acid Ⅰ was determined to be 24.5 μg mg-1. By combining magnetic solid phase extraction with HPLC detection, a method was developed to enrich and detect aristolochic acids used in traditional Chinese medicine. A satisfactory recovery (92.7 - 97.5% for aristolochic acid Ⅰ and 92.6 - 99.4% for aristolochic acid Ⅱ) and an acceptable relative standard deviation (<4.0%) were obtained.


Clinical value of relative quantification ultrasound elastography in characterizing breast tumors.
To evaluate ultrasound elastography (USE) using strain ratio (SR), a relative quantification approach for breast lesions characterization. One hundred forty-seven consecutive patients with a total of 156 breast lesions underwent USE. Technical accuracy was assessed automatically. For SR evaluation a rounded ROI was depicted inside fat (F), glandular tissue (G) and inside the lesion (L), preferably at the same depth. R1, mean value of the G and F ratio, stands for in background tissue composition elasticity. R2; mean value of L/F stands for in lesion elasticity, both evaluated in arbitrary unit (au). Two-years follow-up and pathology results were standard of reference. Mann-Whitney test, ROC analysis and Chi-square with Yates correction were used. With the exception of 27 cysts, 17 malignant and 112 benign lesions were found. R1 values were 1.6±0.7au and 1.2±0.9au (p=0.015 NS); R2 values were 6.1±2.5au and 1.9±1.3au (p<0.001) for malignant and benign lesions, respectively. A threshold of 3.3au showed a sensitivity and specificity of 88% and 87%, respectively with an AUC of 93%. Fifteen false positive and two false negative were detected. Relative quantification of ultrasound elastography allows to find high levels of diagnostic accuracy in characterizing breast tumors above all in downgrading BI-RADS 3 and 4 lesions.


Author Correction: Skeletal muscle energy metabolism during exercise.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Feasibility of a hemodialysis safety checklist for nurses and patients: a quality improvement study.
Patients with end-stage renal disease are at high risk for medical errors given their comorbidities, polypharmacy and coordination of care with other hospital departments. We previously developed a hemodialysis safety checklist (Hemo Pause) to be jointly completed by nurses and patients. Our objective was to determine the feasibility of using this checklist during every hemodialysis session for 3 months. We conducted a single-center, prospective time series study. A convenience sample of 14 nurses and 22 prevalent in-center hemodialysis patients volunteered to participate. All participants were trained in the administration of the Hemo Pause checklist. The primary outcome was completion of the Hemo Pause checklist, which was assessed at weekly intervals. We also measured the acceptability of the Hemo Pause checklist using a local patient safety survey. There were 799 hemodialysis treatments pre-intervention (13 January-5 April 2014) and 757 post-intervention (5 May-26 July 2014). The checklist was completed for 556 of the 757 (73%) treatments. Among the hemodialysis nurses, 93% (13/14) agreed that the checklist was easy to use and 79% (11/14) agreed it should be expanded to other patients. Among the hemodialysis patients, 73% (16/22) agreed that the checklist made them feel safer and should be expanded to other patients. The Hemo Pause safety checklist was acceptable to both nurses and patients over 3 months. Our next step is to spread this checklist locally and conduct a mixed methods study to determine mechanisms by which its use may improve safety culture and reduce adverse events.


Targeting hypoxia to overcome radiation resistance in head & neck cancers: real challenge or clinical fairytale?
Tumor hypoxia is a major cause for failure of therapy in patients with inoperable head and neck cancers. Various anti-hypoxic treatment strategies (e.g. hyperbaric oxygenation, hypoxic cell sensitizers) have been tested in clinical trials in head and neck cancer over the past 30 years and have shown modest improvements in combination with radiotherapy in meta-analyses. Anemia worsens tumor hypoxia, but anemia correction had no significant effect. New approaches (e.g. anti-HIF-directed molecular therapies) have just entered early clinical studies and data are lacking. Expert commentary: A new attractive and promising approach derives from recent advances in imaging and radiotherapy delivery. Progress in imaging of hypoxia (e.g. by positron emission tomography) can select patients for specific therapies and may, in particular, facilitate anti-hypoxia-directed radiotherapy which has become feasible with advanced radiotherapy techniques (IMRT with 'dose-painting'). The combination of both methods may offer a powerful tool for effective targeting of hypoxia in the near future.


De novo molecular subtyping of salivary gland tissue in the context of Sjögren's syndrome heterogeneity.
Understanding the mechanistic features and molecular taxonomy of diseases holds promise for the development of more effective treatments, especially for complex heterogeneous diseases. Here, we analyzed transcriptomic datasets of salivary gland tissues from patients with Sjögren's syndrome (SjS) to identify shared and divergent cellular and molecular signatures. Three molecular subtypes of SjS salivary gland tissue were identified: oxidative phosphorylation (OxPhos)-dominant (C1), weak inflammatory with type I interferon signatures (C2), and B cell receptor (BCR) signaling pathway-dominant (C3). C3 had the highest focus score. Type I helper T cells and B cells were the dominant cell types in C1 and C3 tissues, respectively. Metformin and drugs targeting PI3K, BTK, and JAKs were predicted to be effective treatments for C1 and C3 subtypes, respectively. Three subtypes of SjS salivary gland with distinct molecular signatures were identified. The results could contribute to optimal stratification of patients for more effective treatment approaches.


Bright fluorogenic squaraines with tuned cell entry for selective imaging of plasma membrane vs. endoplasmic reticulum.
A rational design of squaraine dyes with lipophilic and zwitterionic groups tunes cell entry, allowing for selective far-red/near-infrared imaging of plasma membrane vs. endoplasmic reticulum. They exhibit up to 110-fold fluorescence enhancement in biomembranes and enable cellular imaging at 1 nM concentration, which make them the brightest membrane probes to date.


The association of jejunal atresia and cystic lymphangioma in the same location.
Cystic lymphangiomas, which are localized on the mesentery of the intestine, are rarely seen. The presence of intestinal atresia together with cystic lymphangioma in the same location has never been described before. In this study, a premature girl weighing 2,630 g who had jejunal atresia, cystic lymphangioma localized in the mesentery of the distal atretic jejunal segment, and multiple rib anomalies is presented. The association of jejunal atresia and cystic lymphangioma may be coincidental, or jejunal atresia may be caused by an intrauterine vascular compression of volvuled cystic lymphangioma. Such an association is being reported for the first time in the literature.


Influence of saliva contamination and abrasion on resin to tin-plated alloy bond strengths.
This investigation studied the effects of abrasion and salivary contamination on the shear bond strength of a composite resin bonded to tin-plated gold alloy. Experimental groups (n = 15) consisted of: group 1, no surface treatment; group 2, enamel abraded; and group 3, saliva-contaminated. After surface stressing and bonding, groups were thermocycled before shear testing. Fractured interfaces were analyzed by use of light microscopy and SEM. Specimens in group 3 (12.2 +/- 2.5 MPa) had significantly lower bond strengths than group 1 (17.9 +/- 6.8 MPa). The shear bond strengths of resin bonded to tin-plated metal were significantly affected by salivary contamination but not by abrasion.


Contrasting Surgical Management of CSF Leak from Lateral Recess of Sphenoid Sinus and Its Surgical Outcomes: Our Experience.
CSF leak from Lateral Recess of Sphenoid (LRS) sinus accounts for 35% of all CSF rhinorrhoea cases. There are various surgical techniques described for repair of LRS CSF leak. This study describes the experience of LRS leak repair in a tertiary care center with three different surgical techniques. Study comprises of 3 cases of LRS CSF leak that presented to J.S.S. Hospital, during the time period of July 2018-January 2019, who underwent endoscopic CSF leak repair. All three cases underwent endoscopic endonasal transpterygoid approach to the leak site. The closure technique opted for all three cases were different. For the first case free mucosal flap from ipsilateral middle turbinate was used, for the second ipsilateral nasoseptal flap (NSF) was used and contralateral NSF was used for the third. All the cases were followed up for a minimum of 3 months. In all the 3 cases the CSF leak site was located in the lateral recess of Sphenoid sinus. Encephalocele was noted in two cases, which were cauterised and closure was done as planned. Crusting was more in cases that underwent closure using free mucosal flap. Healing and take up was similar for both the ipsilateral NSF and contralateral NSF. The endoscope has revolutionized the management of CSF leak from the lateral recess of sphenoid sinus. These defects can be managed efficiently using multilayer closure of defect. For large defects, the Posterior nasoseptal flaps can be used. In addition, contralateral PNSF has lower chances of being devascularized due to injury to pedicle while drilling the pterygoid plates.


S-IgA cholera toxin and rotavirus antibody in human colostrum.
S-IgA antibodies against cholera toxin and rotavirus were assayed in 43 colostral samples by means of an enzyme-linked immunosorbent assay (ELISA). All specimens contained specific S-IgA antibodies against both antigens. Significant antibody titres to the antigens were demonstrated in almost all colostral samples.


Effect of osmo priming on sucrose metabolism in spring maize, during the period of grain filling, under limited irrigation conditions.
The present study was undertaken to study the effect of osmo priming on sucrose metabolism of spring maize, under limited irrigation conditions. Osmo priming increased the activities of acid invertase, alkaline invertase and sucrose synthase (cleavage) and the contents of reducing sugars and starch in the grains of stressed plants. There was also an increase in sucrose phosphate synthase activity with a parallel increase in sucrose content in leaves of stressed plants in comparison with those of hydro priming treatment. It showed that osmo priming helped in improving sucrose phosphate synthase activity in leaves of plants, leading to higher sucrose content, under stress conditions.


Characterization of extracellular matrix in non-Hodgkin's lymphomas.
Basement membrane proteins laminin, type IV collagen and aminoterminal propeptide of type III collagen (type III pN-collagen) are important components of extracellular matrix of normal lymph nodes; they are present in reticular fibres and in the walls of blood vessels and sinuses. We have studied the distribution of these proteins in 45 cases of non-Hodgkin's lymphomas representing different types of Kiel classification. Light microscopic immunohistochemistry was used on sections from formalin fixed, paraffin embedded biopsy specimens. Reticular cells were identified by using antibodies against S-100 protein, a marker for dendritic and interdigitating reticulum cells, and against cystatin A for the demonstration of follicular dendritic reticulum cells. The findings of the immunohistochemical stainings were generally similar in the different lymphoma types. Thus the method for the demonstration of extracellular matrix proteins could not give obvious aid for lymphoma classification. BM stainings proved to be efficient in the demonstration of sinuses. Marginal and medullary sinuses of the nodal cases were either totally or partly spared and open also in the cases in which there was extracapsular growth. The fibre content was variable, but generally the distribution of all the three extracellular matrix proteins was comparable with each other and mainly corresponded with the pattern seen in conventional Gomori's reticulin stain. This also holds for the tumor tissue infiltrated through the capsule and for extranodal cases. Areas of sclerosis of some cases contained only type III pN-collagen. Reticular fibres were at places seen to be closely associated with cells of spindle or dendritic shape. A few reticulum cells could be found to show cytoplasmic staining for laminin, type IV collagen and type III pN collagen. This finding suggests that the cells are synthetizing these proteins. The amount of cells staining for cystatin A or S-100 protein did not generally correlate with the amount of fibres, and most of the stained cells were not closely associated with fibres. The results show that in non-Hodgkin's lymphomas basement membrane proteins and type III pN-collagen are important components of reticular fibres. Results also indirectly support the main role of fibroblastic reticulum cells in the production of the fibres.


Introductory lecture: from biomolecules to biodiagnostics: spectroscopy does it all.
Optical spectroscopy and imaging have been used in medicine since medicine was first practised. However, the more sophisticated instrumental methods now under development have made little impact on clinical medicine. In this paper a brief overview of the development of optical diagnostics is presented, highlighted by some successful pre-clinical applications. The reasons for the slow penetration of optical diagnostics into clinical practice are discussed.


Complement components (C3, C4) in childhood asthma.
To assess the involvement of complements (C3, C4) in the pathophysiology of bronchial asthma. Selection of patients (n = 64) were made according to the recommended international criteria for diagnosis and classification of asthma. Serum levels of complement components (C3, C4) were measured by radial immunodiffusion technique in 64 Libyan children (age: 1-12 years, sex: 39 males, 25 females) with mild to moderately severe asthma (Group A). Among these patients, 35 had active disease (AA) and 29 had inactive disease (NA). According to age range, 20, 21 and 23 patients were between 1-3 years (A1), > 3-5 years (A2) and > 5-12 years (A3) respectively. A1 had 9 and 11 patients with active (AA1) and inactive (NA1) disease; A2 had 10 and 11 patients with active (AA2) and inactive (NA2) disease; A3 had 16 and 7 patients with active (AA3) and inactive (NA3) disease respectively. Age matched comparisons were made with 57 healthy children (age: 1-12 years; sex: 30 males, 27 females) (Group B). Among the controls, 15, 19 and 23 children were between 1-3 years (B1), > 3-5 years (B2) and > 5-12 years (B3) respectively. Mean C3 level was significantly elevated in patients, while C4 level was normal (A vs B --> C3: P < 0.2, C4: P > 0.2). Serum C3 level was significantly higher in patients with active disease only, while it was normal in patients with inactive disease (AA, NA, B --> P = 0.045); AA vs NA --> P < 0.05, AA vs B --> P < 0.02, NA vs B --> P > 0.05) and C4 levels were normal in both the groups (AA, NA, B --> P = 0.354). Further, C3 levels were significantly elevated in all the age groups, but in patients with active disease only (AA1, NA1, B1 --> P = 0.0024; AA2, NA2, B2 --> P = 0.0411; AA3, NA3, B3 --> P = 0.0102). The elevated C3 level was possibly due to induction by pro-inflammatory cytokines such as tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1). The probable mechanisms of C3 involvement in the pathophysiology of bronchial asthma were discussed.


Internal sphincterotomy versus topical nitroglycerin ointment for chronic anal fissure.
Anal fissure is a common benign condition. An anorectal problem is defined as a split in the anal canal mucosa that extends from the dentate line to the anal verge. Chronic anal fissure is defined by a history of symptoms present for more than 2 months' duration and with a triad of external skin tags, namely, a hypertrophied anal papilla, an ulcer with rolled edges, and a base exposing the internal sphincter. Because complications such as incontinence are associated with surgical treatment, chemical sphincterotomy is currently favored. The objective of this study is to compare the difference in outcome between open partial lateral anal sphincterotomy and application of topical 0.2% nitroglycerin ointment for the treatment of chronic anal fissure. This was a quasi-experimental study carried out between January 16, 2007 and January 15, 2008 in the Surgical Department of Jinnah Hospital, Lahore, Pakistan. Sixty consecutive cases with a clinical diagnosis of chronic anal fissure were recruited in the study. All recruited patients met the study inclusion criteria and were randomly assigned to one of the two groups. Group A was managed conservatively using topical 0.2% nitroglycerin ointment, whereas Group B underwent open partial lateral anal sphincterotomy. Both groups were followed up at 1 week, 2 weeks, 4 weeks, and 6 weeks after the treatment. All the patients complained of pain. A total of 43 (71.7%) patients had pain with constipation, whereas 31 (51.7%) patients had bleeding per rectum. Upon clinically examining the anal area, tenderness was elicited in all 60 (100%) patients. Group A included 30 (11 females and 19 males) cases treated with topical 0.2% nitroglycerin ointment and Group B included 30 (11 females and 19 males) cases who underwent open partial lateral anal sphincterotomy. In Group A, only 15 patients with fissures were successfully treated (50%). By contrast, 28 (93%) patients with fissures in Group B were successfully treated, and only two (7%) remained uncured. These two patients (6.6%) in Group B suffered from incontinence due to flatus and feces as a complication of the procedure. This quasi-experimental study demonstrates that open partial lateral internal sphincterotomy is superior to topical 0.2% nitroglycerin application in the treatment of chronic anal fissure, with good symptomatic relief, high rate of healing, fewer side effects, and a very low rate of early continence disturbances.


Comparison of intraosseous and intravenous routes of anticonvulsant administration in a porcine model.
During status epilepticus, rapid IV access for the administration of anticonvulsant drugs can be a very difficult and time-consuming procedure. Our study evaluated whether therapeutic serum levels of phenobarbital and phenytoin could be obtained by the intraosseous route. Twenty domestic swine weighing 10 to 20 kg were divided into two groups (ten each). In one group, phenobarbital 20 mg/kg was administered either intravenously (five) or intraosseously (five). The second group received phenytoin 15 mg/kg either intravenously (five) or intraosseously (five). All animals had samples for anticonvulsant levels drawn from an indwelling arterial cannula at one, three, five, seven, ten, 15, and 30 minutes after dosing. Anticonvulsant levels were found to be statistically significantly higher with IV administration (P less than .01). However, phenobarbital levels were therapeutic by the intraosseous route, while phenytoin levels were below the therapeutic range after the ten-minute interval. Bone marrow levels 45 minutes after infusion were 13.5 micrograms/mL (phenobarbital) and 11.5 micrograms/mL (phenytoin). Our study demonstrates that current IV dosing of phenobarbital 20 mg/kg given intraosseously obtains and maintains therapeutic serum levels. Phenytoin 15 mg/kg does not maintain therapeutic levels and cannot be recommended for intraosseous administration.


Revealing the biexciton and trion-exciton complexes in BN encapsulated WSe2.
Strong Coulomb interactions in single-layer transition metal dichalcogenides (TMDs) result in the emergence of strongly bound excitons, trions, and biexcitons. These excitonic complexes possess the valley degree of freedom, which can be exploited for quantum optoelectronics. However, in contrast to the good understanding of the exciton and trion properties, the binding energy of the biexciton remains elusive, with theoretical calculations and experimental studies reporting discrepant results. In this work, we resolve the conflict by employing low-temperature photoluminescence spectroscopy to identify the biexciton state in BN-encapsulated single-layer WSe2. The biexciton state only exists in charge-neutral WSe2, which is realized through the control of efficient electrostatic gating. In the lightly electron-doped WSe2, one free electron binds to a biexciton and forms the trion-exciton complex. Improved understanding of the biexciton and trion-exciton complexes paves the way for exploiting the many-body physics in TMDs for novel optoelectronics applications.


Carcinoma of the fallopian tube.
Carcinoma of the fallopian tube is the least frequent tumor of the female genital tract. It may occur at any age but is more common in the sixth decade. The tumor involves both tubes in 26 per cent of cases. The spread is by direct extension to peritoneum. The symptoms include metrorrhagia, leukorrhea, abdominal pain, singly or in combination. The diagnosis is difficult but could be made more frequently if the causes of abnormal bleeding were thoroughly investigated by means of cytology and endometrial curettage. Treatment is by resection of the tumor, total hysterectomy, and bilateral salpingo-oophorectomy followed by chemotherapy. The 5-year survival rate is 38 per cent.


Structure investigation of intramolecular hydrogen bond in some substituted salicylaldehydes and 4-aminoantipyrine derivatives in solution and in the solid state.
Seven imine derivatives obtained by condensation of appropriate aldehydes and salicylaldehydes with 4-aminoantipyrine were investigated in terms of intramolecular hydrogen bond structure. On the base of (1)H, (13)C and (15)N NMR measurements in solution and in the solid state we found out that all compounds which can form such structure exist as OH forms with strong H-bonds to nitrogen atom. The structure conclusions taken from NMR study were confirmed by pKa measurements. Surpassingly, the positions of protons in H-bridges only very slightly depend on the substituents in aldehyde used for condensation and on the phase (solution vs. solid state). The influence of antipyrine moiety seems to be the major factor defining H-bond structure.


Rh(III)-Catalyzed One-Step Synthesis of ortho-Alkynylated Perylene Imide Dyes: Optical and Electrochemical Properties of New Derivatives.
A one-step Rh-catalyzed site-selective ortho-C-H alkynylation of perylene as well as naphthalene mono- and diimides is reported. A single step regioselective access to ortho-C-H alkynylated derivatives of these ryleneimides not only increases the step economy of the ortho-functionalization on these dyes but also provides a quick access route towards highly functionalized dyes that have potential optoelectronic applications. Increased solubility of tetra(triisopropylsilyl)acetylenyl PDIs in organic solvents greatly enhances their utility for further derivatization.


Poor Patient Compliance Limits the Attainability of Patient-Reported Outcome Measure Completion Thresholds for the Comprehensive Care for Joint Arthroplasty Model.
The Centers for Medicare and Medicaid Services (CMS) links patient-reported outcome measures (PROMs) with hospital reimbursement in some value-based models for total joint arthroplasty (TJA). This study evaluates PROM reporting compliance and resource utilization using protocol-driven electronic collection of outcomes for commercial and CMS alternative payment models (APMs). We analyzed a consecutive series of patients undergoing total hip arthroplasty (THA) and total knee arthroplasty (TKA) from 2016 and 2019. Compliance rates were obtained for reporting hip disability and osteoarthritis outcome score for joint replacement (HOOS-JR.), knee disability and osteoarthritis outcome score for joint replacement (KOOS-JR.), and 12-item short form survey (SF-12) surveys preoperatively and postoperatively at 6-months, 1 year, and 2- years. Of 43,252 THA and TKA patients, 25,315 (58%) were Medicare-only. Direct supply and staff labor costs for PROM collection were obtained. Chi-square testing compared compliance rates between Medicare-only and all-arthroplasty groups. Time-driven activity-based costing (TDABC) estimated resource utilization for PROM collection. In the Medicare-only cohort, preoperative HOOS-JR./KOOS-JR. compliance was 66.6%. Postoperative HOOS-JR./KOOS-JR. compliance was 29.9%, 46.1%, and 27.8% at 6 months, 1 year, and 2 years, respectively. Preoperative SF-12 compliance was 70%. Postoperative SF-12 compliance was 35.9%, 49.6%, and 33.4% at 6 months, 1 year, and 2 years, respectively. Medicare patients had lower PROM compliance than the overall cohort (P < .05) at all time points except preoperative KOOS-JR., HOOS-JR., and SF-12 in TKA patients. The estimated annual cost for PROM collection was $273,682 and the total cost for the entire study period was $986,369. Despite extensive experience with APMs and a total expenditure near $1,000,000, our center demonstrated low preoperative and postoperative PROM compliance rates. In order for practices to achieve satisfactory compliance, Comprehensive Care for Joint Replacement (CJR) compensation should be adjusted to reflect the costs associated with collecting these PROMs and CJR target compliance rates should be adjusted to reflect more attainable levels consistent with currently published literature.


Versatile properties of an exopolysaccharide R-PS18 produced by Rhizobium sp. PRIM-18.
Exopolysaccharides (EPS) produced by bacteria have attracted scientific and industrial attention due to their multifunctional properties and relatively easier production. In this study, an EPS viz., R-PS18 produced by Rhizobium sp. PRIM-18 was characterized and its functional properties were assessed. Cell proliferative and in vitro wound healing activities of the EPS were established using human dermal fibroblast (HDF) cells. The isolate produced 2.1 g L(-1) purified EPS (molecular weight 9.33×10(6) Da) comprising of glucose, galactose, and mannose (6.1:1.8:1). Viscosity of 0.25% solution was 23.4 mPa s (shear rate 75 s(-1)) and it showed pseudoplastic and thixotropic behavior. High emulsification, iron chelation, and superoxide scavenging abilities were also observed. Significant increase in HDF cell proliferation and wound healing in vitro was achieved by R-PS18 treatment. Sulfation of R-PS18 significantly enhanced the cell proliferative and wound healing activities. In conclusion, these findings indicate potential applications of R-PS18.


Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos.
Human embryonic stem (hES) cells are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. Most hES cell lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in excess of clinical need. Embryos that are discarded during the IVF procedure because of poor morphology and a low likelihood for generating viable pregnancies or surviving the cryopreservation process are also a viable source of hES cells. In this protocol, we describe how to derive novel hES cells from discarded poor-quality embryos and how to maintain the hES cell lines.


Chromatin interaction-aware gene regulatory modeling with graph attention networks.
Linking distal enhancers to genes and modeling their impact on target gene expression are longstanding unresolved problems in regulatory genomics and critical for interpreting noncoding genetic variation. Here, we present a new deep learning approach called GraphReg that exploits 3D interactions from chromosome conformation capture assays to predict gene expression from 1D epigenomic data or genomic DNA sequence. By using graph attention networks to exploit the connectivity of distal elements up to 2 Mb away in the genome, GraphReg more faithfully models gene regulation and more accurately predicts gene expression levels than the state-of-the-art deep learning methods for this task. Feature attribution used with GraphReg accurately identifies functional enhancers of genes, as validated by CRISPRi-FlowFISH and TAP-seq assays, outperforming both convolutional neural networks (CNNs) and the recently proposed activity-by-contact model. Sequence-based GraphReg also accurately predicts direct transcription factor (TF) targets as validated by CRISPRi TF knockout experiments via in silico ablation of TF binding motifs. GraphReg therefore represents an important advance in modeling the regulatory impact of epigenomic and sequence elements.


Clinical trial of the efficacy and safety of ticarcillin and clavulanic acid.
Forty-three hospitalized patients were treated with a new antibiotic combination containing ticarcillin plus the beta-lactamase inhibitor, clavulanic acid, in a fixed combination for intravenous use. A variety of infections were treated, including pneumonia, bacteremia, urinary tract infection, and osteomyelitis. Of 50 episodes of infection in 43 patients, 44 clinical cures were obtained, with 5 patients improving and 1 patient failing to respond to treatment. In vitro susceptibility testing of 101 clinical isolates was notable for the rarity of resistance to the combination antibiotic. Of specific interest, all 14 isolates of Staphylococcus aureus were susceptible to ticarcillin plus clavulanic acid, whereas only 2 of the 14 isolates were susceptible to ticarcillin alone. Adverse reactions to the study drug were minimal; eosinophilia, unaccompanied by other allergic phenomena, and oral candidiasis were most frequent. Overall, the combination of ticarcillin with the beta-lactamase inhibitor, clavulanic acid, appears to be a safe and effective drug for the treatment of infections caused by susceptible organisms.


Upregulation of Dickkopf1 by oscillatory shear stress accelerates atherogenesis.
Numerous clinical studies have highlighted the pivotal role Dickkopf (DKK) 1 plays in atherosclerosis, but the underlying mechanisms remain unknown. The present study was designed to explore the contribution of DKK1 to the development of atherosclerosis under oscillatory shear stress. Oscillatory shear stress applied to endothelial cells induced DKK1 expression, which peaked at 6 h. siRNA knockdown or silencing DKK1 by lentiviral gene delivery counteracted the increased monocyte adhesion and impaired endothelial tight junction induced by oscillatory shear stress, thereby attenuating atherogenesis in ApoE-/- mice. As well, activation of endothelial proteinase-activated receptor 1 (PAR1) and its downstream transcription factor, cAMP response element-binding protein (CREB), was critical to the increased expression of DKK1 under oscillatory shear stress. We provide evidence that DKK1 contributes to the development of atherosclerosis under conditions of oscillatory shear stress. A better understanding of the role played by DKK1 in atherogenesis may provide clinicians with opportunities to prevent atherosclerosis. Disturbed oscillatory flow increases DKK1 expression. DKK1 knockdown attenuates OSS-induced monocyte adhesion and endothelial impairment. Genetic silencing of DKK1 limits atherogenesis in ApoE-/- mice. Activation of the PAR1/CREB pathway contributes to the upregulation of DKK1 via OSS. DKK1 is a promising candidate with respect to the treatment of atherosclerosis.


Topological localization in out-of-equilibrium dissipative systems.
In this paper, we report that notions of topological protection can be applied to stationary configurations that are driven far from equilibrium by active, dissipative processes. We consider two physically disparate systems: stochastic networks governed by microscopic single-particle dynamics, and collections of driven interacting particles described by coarse-grained hydrodynamic theory. We derive our results by mapping to well-known electronic models and exploiting the resulting correspondence between a bulk topological number and the spectrum of dissipative modes localized at the boundary. For the Markov networks, we report a general procedure to uncover the topological properties in terms of the transition rates. For the active fluid on a substrate, we introduce a topological interpretation of fluid dissipative modes at the edge. In both cases, the presence of dissipative couplings to the environment that break time-reversal symmetry are crucial to ensuring topological protection. These examples constitute proof of principle that notions of topological protection do indeed extend to dissipative processes operating out of equilibrium. Such topologically robust boundary modes have implications for both biological and synthetic systems.


Association of Cost Savings and Surgical Quality With Single-Vendor Procurement for Spinal Implants.
Significant cost savings can be achieved from consolidating purchases of spinal implants with a single vendor. However, it is currently unknown whether sole-source contracting or vendor rationalization more broadly affects patient care. To describe the single-vendor procurement of spinal implants, characterize the economic benefits of sole-source contracting, and gauge whether vendor rationalization is associated with a diminished quality of care. This retrospective cohort study assessed adult patients receiving single-level lumbar interbody fusions at a single institution from January 1, 2009, to July 31, 2017. Exclusion criteria included multilevel fusions and prior spinal fusions. Patients were analyzed based on the number of vendors available to surgeons at the time of the patient's surgery. January 1, 2009, to December 31, 2010, was defined as the multivendor period (10 vendors); January 1, 2011, to December 31, 2014, was defined as the dual-vendor period; and January 1, 2015, to July 31, 2017, was defined as the single-vendor period. Rates of 12-month revision surgery, complications, 30-day readmissions, and postoperative patient-reported outcomes, as measured by 5-dimension European Quality of Life (EQ-5D) and Patient-Reported Outcomes Measurement Information System-Global Health (PROMIS-GH) utilities. Propensity score weighting was performed to control for confounding. The Holm method was used to correct for multiple testing. Annual cost savings associated with the dual-vendor and single-vendor periods were also reported. A total of 1373 patients (mean [SD] age, 59.2 [12.6] years; 763 [55.6%] female; 1161 [84.6%] white) were analyzed. Rates of revisions after adjusting for confounding were 3.2% (95% CI, 1.5%-6.7%) for the multivendor period, 4.5% (95% CI, 3.1%-6.5%) for the dual-vendor period, and 3.0% (95% CI, 1.7%-5.0%) for the single-vendor period. Complication rates were 5.3% (95% CI, 2.7%-10.1%) for the multivendor period, 7.2% (95% CI, 5.4%-9.6%) for the dual-vendor period, and 6.4% (95% CI, 4.6%-8.8%) for the single-vendor period. Readmission rates were 14.2% (95% CI, 9.7%-20.2%) for the multivendor period, 12.6% (95% CI, 10.1%-15.5%) for the dual-vendor period, and 9.7% (95% CI, 7.4%-12.7%) for the single-vendor period. Revisions, complications, and patient-reported outcomes were statistically equivalent across all periods. Readmissions were not statistically equivalent but not statistically different. The savings attributable to vendor rationalization were 24% for the dual-vendor and 21% for the single-vendor periods. The single-vendor procurement of spinal implants was associated with significant cost savings without evidence of an associated decline in the quality of care. Large hospital systems may consider sole-source purchasing as a viable cost reduction strategy.


Correction to: Nomenclature report 2019: major histocompatibility complex genes and alleles of great and small ape and old and new world monkey species.
The original version of this article contained a spelling error in the Acknowledgments regarding the name of the funding organisation supporting GM and JAH. UKRI-BBSCR should have been UKRI-BBSRC, as is now indicated correctly below.


With free health services, why does the Brazilian working class delay in seeing the doctor?
The length of delay in seeing a physician was related to perceived difficulty in access to services and to scepticism and doubt about the effectiveness of medical care. Attention is called to the importance of the doctor-patient relationship in fostering prompt patient action.


Validation of an instrument to determine oral health knowledge, attitudes, and practices during pregnancy.
Oral health of the mother-infant dyad is important to preserve general health. However, there are few instruments in Spanish for the evaluation of knowledge, attitudes and practices that determine this construct. Therefore, this research aimed to develop and evaluate the psychometric properties of the Maternal Oral Health Knowledge, Attitudes and Practices Questionnaire (CAPSOM in Spanish). In this instrument development study that carried out in 2018-2019, involving pregnant women between the ages of 18 and 45 in the city of Leon, Guanajuato, Mexico. The sample size was calculated based on 10 women per questionnaire item (n = 10 k). The study used Cronbach's alpha, the modified Lawshe test of validity criteria, factor analysis, and the level of difficulty and discrimination of the items. 207 women took part with their signed, informed consent (25 ± 6 years). The internal consistency of the instrument, both total and by dimension was α = 0.70, α = 0.66 knowledge, α = 0.74 attitudes, and α = 0.66 practices. Values of Content Validity Ratio' ≥ 0.60 were obtained for the final 10 items and Content Validity Index' = 0.90. The average difficulty index of items was 0.40, and there were significant differences (Kruskall-Wallis, p < 0.001) in the discrimination test. Factor analysis demonstrated three main components. A valid and reliable 10-item Spanish questionnaire was designed to measure pregnant women's oral health knowledge, attitudes, and practices.


Gold(I)-catalyzed divergence in the reactivity of 3-silyloxy 1,6-enynes: pinacol-terminated vs claisen-terminated cyclization cascades.
On activation with catalytic amounts of gold(I) complexes, 3-silyloxy 1,6-enynes can react through two alternative pathways. In one, a cascade reaction consisting of carbocyclization and subsequent pinacol rearrangement takes place. In the second pathway, a heterocyclization is followed by a Claisen rearrangement. The reaction outcome differs depending on the substitution pattern of the 3-silyloxy 1,6-enynes and, more importantly, the electronic properties of the gold-bound phosphane ligand.


Role of molecular biomarkers in glioma resection: a systematic review.
New discoveries based on genetic and epigenetic evidence have significantly expanded the understanding of diffuse gliomas. Molecular biomarkers detected in diffuse gliomas are not only potential targets for radiotherapy, chemotherapy, and immunotherapy, but are also able to guide surgical treatment. Previous studies have suggested that the optimal extent of resection of diffuse gliomas varies according to the expression of specific molecular biomarkers. However, the specific guiding role of these biomarkers in the resection of diffuse gliomas has not been systemically analyzed. This review summarizes several critical molecular biomarkers of tumorigenesis and progression in diffuse gliomas and discusses different strategies of tumor resection in the context of varying genetic expression. With ongoing study and advances in technology, molecular biomarkers will play a more important role in glioma resection and maximize the survival benefit from surgery for diffuse gliomas.


Injectable click-crosslinked hydrogel containing resveratrol to improve the therapeutic effect in triple negative breast cancer.
Triple-negative breast cancer (TNBC) patients are considered intractable, as this disease has few effective treatments and a very poor prognosis even in its early stages. Here, intratumoral therapy with resveratrol (Res), which has anticancer and metastasis inhibitory effects, was proposed for the effective treatment of TNBC. An injectable Res-loaded click-crosslinked hyaluronic acid (Res-Cx-HA) hydrogel was designed and intratumorally injected to generate a Res-Cx-HA depot inside the tumor. The Res-Cx-HA formulation exhibited good injectability into the tumor tissue, quick depot formation inside the tumor, and the depot remained inside the injected tumor for extended periods. In vivo formed Res-Cx-HA depots sustained Res inside the tumor for extended periods. More importantly, the bioavailability and therapeutic efficacy of Res remained almost exclusively within the tumor and not in other organs. Intratumoral injection of Res-Cx-HA in animal models resulted in significant negative tumor growth rates (i.e., the tumor volume decreased over time) coupled with large apoptotic cells and limited angiogenesis in tumors. Therefore, Res-Cx-HA intratumoral injection is a promising way to treat TNBC patients with high efficacy and minimal adverse effects.


Survival outcomes for patients with localised upper tract urothelial carcinoma managed with non-definitive treatment.
To investigate the outcomes of patients with upper tract urothelial carcinoma (UTUC) with non-definitive therapy, which currently remains unknown. We used the Surveillance, Epidemiology, and End Results (SEER) database to identify individuals with a localised, histologically confirmed kidney/renal pelvis and ureteric UC. Survival analysis using the Kaplan-Meier method was performed. A competing risk model evaluated the cumulative incidence and predictors of cancer-specific mortality (CSM). We identified 633 (7.6%) individuals who did not receive surgery. These individuals were significantly older (median age 81 vs 71 years, P < 0.001) than surgically managed patients. The median overall survival (OS) was significantly shorter compared to the surgical cohort (1.9 vs 7.8 years, P < 0.001). The 3-year disease-specific survival (DSS) for patients without surgery was significantly lower compared to those with surgery, at 73.7% vs 92.4%, respectively (P < 0.001). The 3-year DSS for patients with high-grade tumours was worse when compared to patients with low-grade tumours, at 65.1% vs 82.9%, respectively (P < 0.001). The 3-year cumulative CSM was 26.3%. On multivariable analysis, older age (hazard ratio [HR] 1.05, P < 0.001) and high tumour grade (HR 1.88, P < 0.001) were predictors of worse outcomes. In this population-based cohort, 7.6% of patients with UTUC were managed with a non-definitive approach. The median OS for the untreated cohort was significantly shorter compared to the surgical cohort (1.9 vs 7.8 years, respectively). These data may be helpful in counselling patients who are poor surgical candidates, as non-definitive therapy may provide reasonable oncological outcomes.


Differential proteome analysis of hagfish dental and somatic skeletal muscles.
Hagfish, the plesiomorphic sister group of all vertebrates, are deep-sea scavengers. The large musculus (m.) longitudinalis linguae (dental muscle) is a specialized element of the feeding apparatus that facilitates the efficient ingestion of food. In this article, we compare the protein expression in hagfish dental and somatic (the m. parietalis) skeletal muscles via two-dimensional gel electrophoresis and mass spectrometry in order to characterize the former muscle. Of the 500 proteins screened, 24 were identified with significant differential expression between these muscles. The proteins that were overexpressed in the dental muscle compared to the somatic muscle were troponin C (TnC), glycogen phosphorylase, beta-enolase, fructose-bisphosphate aldolase A (aldolase A), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In contrast, myosin light chain 1 (MLC 1) and creatine kinase (CK) were over-expressed in the somatic muscle relative to the dental muscle. These results suggest that these two muscles have different energy sources and contractile properties and provide an initial representative map for comparative studies of muscle-protein expression in low craniates.


Simplified quality assurance of floor model centrifuges.
Present methods for calibration of the rotor speed of large floor model centrifuges used for blood component preparation can be cumbersome and/or dangerous. This report describes a safe and inexpensive method of modifying a centrifuge lid so that calibration of the machine with a photoelectric tachometer is a simple and safe procedure.


Decoding the prognostic significance of integrator complex subunit 9 (INTS9) in glioma: links to TP53 mutations, E2F signaling, and inflammatory microenvironments.
Gliomas, a type of brain neoplasm, are prevalent and often fatal. Molecular diagnostics have improved understanding, but treatment options are limited. This study investigates the role of INTS9 in processing small nuclear RNA (snRNA), which is crucial to generating mature messenger RNA (mRNA). We aim to employ advanced bioinformatics analyses with large-scale databases and conduct functional experiments to elucidate its potential role in glioma therapeutics. We collected genomic, proteomic, and Whole-Exon-Sequencing data from The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) for bioinformatic analyses. Then, we validated INTS9 protein expression through immunohistochemistry and assessed its correlation with P53 and KI67 protein expression. Gene Set Enrichment Analysis (GSEA) was performed to identify altered signaling pathways, and functional experiments were conducted on three cell lines treated with siINTS9. Then, we also investigate the impacts of tumor heterogeneity on INTS9 expression by integrating single-cell sequencing, 12-cell state prediction, and CIBERSORT analyses. Finally, we also observed longitudinal changes in INTS9 using the Glioma Longitudinal Analysis (GLASS) dataset. Our findings showed increased INTS9 levels in tumor tissue compared to non-neoplastic components, correlating with high tumor grading and proliferation index. TP53 mutation was the most notable factor associated with upregulated INTS9, along with other potential contributors, such as combined chromosome 7 gain/10 loss, TERT promoter mutation, and increased Tumor Mutational Burden (TMB). In GSEA analyses, we also linked INTS9 with enhanced cell proliferation and inflammation signaling. Downregulating INTS9 impacted cellular proliferation and cell cycle regulation during the function validation. In the context of the 12 cell states, INTS9 correlated with tumor-stem and tumor-proliferative-stem cells. CIBERSORT analyses revealed increased INTS9 associated with increased macrophage M0 and M2 but depletion of monocytes. Longitudinally, we also noticed that the INTS9 expression declined during recurrence in IDH wildtype. This study assessed the role of INTS9 protein in glioma development and its potential as a therapeutic target. Results indicated elevated INTS9 levels were linked to increased proliferation capacity, higher tumor grading, and poorer prognosis, potentially resulting from TP53 mutations. This research highlights the potential of INTS9 as a promising target for glioma treatment.


Youth-Led Participatory Action Research: Developmental and Equity Perspectives.
Youth-led participatory action research (YPAR) is an approach to scientific inquiry and social change grounded in principles of equity that engages young people in identifying problems relevant to their own lives, conducting research to understand the problems, and advocating for changes based on research evidence. This chapter provides an introduction to YPAR followed by consideration of the (a) developmental relevance of YPAR for marginalized youth, (b) implications of YPAR for developmental science research on inequities experienced by youth, and (c) potential opportunities and impact of YPAR for improving key developmental settings such as schools and youth-serving organizations. Resources for conducting YPAR projects are discussed, as well as the need for potential integration of YPAR and other participatory approaches to engaging youth and their expertise-at a significant enough scale to have a meaningful impact on policies and practices that affect youth development.


A new hydrogel intraocular lens design.
A new hydrogel intraocular lens was designed to improve fixation and support. The lens, IOGEL model 2000S, has a 6.00 mm optic that merges via a crescentic flange into a terminal loop for an overall diameter of 12.00 mm. The haptics are more flexible than existing loops and the design of the lens is based on a concept of minimum loop rigidity; that is, a haptic that is rigid enough to support the weight of a lens in aqueous within the capsular bag is considered adequate for modern cataract surgery. I evaluated the lens design's safety and efficacy. Excluding patients with pre-existing macular degeneration, the corrected acuity of all patients at last follow-up was 20/40 or better in 100%, 20/30 or better in 95%, 20/25 to 20/20 or better in 85%, and 20/15 or better in 15%. The new haptic can accommodate compression of the capsular bag and allows early, reliable fixation. The centration of the lens was excellent early in the postoperative period and later after capsular fibrosis developed.


Genomic targets and selective inhibition of DNA methyltransferase isoforms.
DNA methylation in the human genome is established and maintained by DNA methyltransferases (DNMTs). DNMT isoforms show differential expression by cell lineage and during development, but much remains to be elucidated about their shared and unique genomic targets. We examined changes in the epigenome following overexpression of 13 DNMT isoforms in HEK293T cells. We observed increased methylation (Δβ > 0.2) at 43,405 CpG sites, with expression of DNMT3A2, DNMTΔ3B4 and DNMTΔ3B2 associated with the greatest impact. De novo methylation occurred primarily within open sea regions and at loci with intermediate methylation levels (β: 0.2-0.6). 53% of differentially methylated loci showed specificity towards a single DNMT subfamily, primarily DNMTΔ3B and DNMT3A and 39% towards a single isoform. These loci were significantly enriched for pathways related to neuronal development (DNMTΔ3B4), calcium homeostasis (DNMTΔ3B3) and ion transport (DNMT3L). Repetitive elements did not display differential sensitivity to overexpressed DNMTs, but hypermethylation of Alu elements was associated with their evolutionary age following overexpression of DNMT3A2, DNMT3B1, DNMT3B2 and DNMT3L. Differential methylation (Δβ > 0.1) was observed at 121 of the 353 loci associated with the Horvath 'epigenetic clock' model of ageing, with 51 showing isoform specificity, and was associated with reduction of epigenetic age by 5-15 years following overexpression of seven isoforms. Finally, we demonstrate the potential for dietary constituents to modify epigenetic marks through isoform-specific inhibition of methylation activity. Our results provide insight into regions of the genome methylated uniquely by specific DNMT isoforms and demonstrate the potential for dietary intervention to modify the epigenome.


Computer-aided diagnosis of pectus excavatum using CT images and deep learning methods.
Pectus excavatum (PE) is one of the most common chest wall defects. Accurate assessment of PE deformities is critical for effective surgical intervention. Index-based evaluations have become the standard for objectively estimating PE, however, these indexes cannot represent the whole information of chest CT images and may associated with significant error due to the individual differences. To overcome these limitations, this paper developed a computer-aided diagnosis (CAD) system based on the convolutional neural network (CNN) to automatically learn discriminative features and classify PE images. We also adopted block-wise fine-tuning methods based on the transfer learning strategy to reduce the potential risk of overfitting caused by limited data and experimentally explored the best fine-tuning degree. Our method achieved a high level of classification accuracy with 94.76% for PE diagnosis. Furthermore, we proposed a majority rule-based voting method to provide a comprehensively diagnostic results for each patient, which integrated the classification results of the whole thorax. The promising results support the feasibility of our proposed CNN-based CAD system for automatic PE diagnosis, which paves a way for comprehensive assessments of PE in clinics.


Bleomycin infusion followed by cyclophosphamide, methotrexate, and 5-fluorouracil in advanced squamous carcinoma of the head and neck.
Twenty-seven patients with squamous cell carcinoma of the head and neck were treated with bleomycin 7.5 U/m2/day continuous infusion X 3 days (days 1-4, 8-11), followed by cyclophosphamide 300 mg/m2, methotrexate 30 mg/m2 and 5-fluorouracil 300 mg/m2 on days 5 and 12 (bleo-CMF). Treatment was repeated every 28 days. All but two had a performance status of 0-2 and all but 3 patients had received prior surgery, radiotherapy, and/or chemotherapy. Of 24 evaluable patients, 1 had a complete remission (1.9 months) and 4 had partial remissions (3.7, 3.9, 3.9, 4.1 months, respectively) for an overall response rate of 21%. If one excludes 7 patients with prior chemotherapy, the response rate is 5 of 17 (29%). All responders had received both radiotherapy and surgery. The median survival was 4.4 months for the responders and 3.5 months for the nonresponders. Marked hematologic toxicity occurred in eight patients, and contributed to the death of two. Severe pulmonary toxicity occurred in two patients and caused the death of one. Bleo-CMF did not produce a higher response rate than historical single-agent trials and caused significant toxicity. Furthermore, there was no important difference in the survival of responders and nonresponders.


The effect of the Fermi resonance on the Raman scattering cross sections of the Fermi doublet ν1 and 2ν2 of liquid carbon disulfide in benzene.
The effect of the Fermi resonance (FR) on the Raman scattering cross sections (RSCSs) of the Fermi doublet ν1, 2ν2 of liquid CS2 in C6H6 using the method of changing the volume concentration of the solution is investigated. We have calculated the RSCSs of the Fermi doublet ν1, 2ν2 using Onsager's theory with the 992 cm(-1) Raman line of C6H6 as the internal standard. The result shows that the RSCS of the ν1 line decreases with decreasing the volume concentration of CS2, while that of the 2ν2 line unexpectedly increases. With decreasing the volume concentration of CS2, two main effects of the solvent effect (SE) and the FR in binary solution that can make the ν1, 2ν2 RSCSs change: the SE, as calculated, reduces both the ν1 and 2ν2 RSCSs; the FR plays a significant role in reducing the ν1 RSCS and enhancing the 2ν2 RSCS. In comparison with our previous investigation [J. Raman Spectrosc. 41 (2010) 776-779], it was found that the stronger the FR is, the more the RSCS of the ν1 decreases and the 2ν2 increases. Thus, we proposed that the result can be best explained by taking into account the effect of the FR on the RSCSs of the Fermi doublet. In addition, this paper also gives an explanation to the experimental results deviating from the theoretical results of the scattering coefficients of CS2 in solvent C6H6 as mentioned in Fini's paper.


Efflux-linked accelerated evolution of antibiotic resistance at a population edge.
Efflux is a common mechanism of resistance to antibiotics. We show that efflux itself promotes accumulation of antibiotic-resistance mutations (ARMs). This phenomenon was initially discovered in a bacterial swarm where the linked phenotypes of high efflux and high mutation frequencies spatially segregated to the edge, driven there by motility. We have uncovered and validated a global regulatory network connecting high efflux to downregulation of specific DNA-repair pathways even in non-swarming states. The efflux-DNA repair link was corroborated in a clinical "resistome" database: genomes with mutations that increase efflux exhibit a significant increase in ARMs. Accordingly, efflux inhibitors decreased evolvability to antibiotic resistance. Swarms also revealed how bacterial populations serve as a reservoir of ARMs even in the absence of antibiotic selection pressure. High efflux at the edge births mutants that, despite compromised fitness, survive there because of reduced competition. This finding is relevant to biofilms where efflux activity is high.


An Innovative Concept for a Multivariate Plausibility Assessment of Simultaneously Recorded Data.
The aim of this work was to develop an innovative multivariate plausibility assessment (MPA) algorithm in order to differentiate between 'physiologically normal', 'physiologically extreme' and 'implausible' observations in simultaneously recorded data. The underlying concept is based on the fact that different measurable parameters are often physiologically linked. If physiologically extreme observations occur due to disease, incident or hormonal cycles, usually more than one measurable trait is affected. In contrast, extreme values of a single trait are most likely implausible if all other traits show values in a normal range. For demonstration purposes, the MPA was applied on a time series data set which was collected on 100 cows in 10 commercial dairy farms. Continuous measurements comprised climate data, intra-reticular pH and temperature, jaw movement and locomotion behavior. Non-continuous measurements included milk yield, milk components, milk mid-infrared spectra and blood parameters. After the application of the MPA, in particular the pH data showed the most implausible observations with approximately 5% of the measured values. The other traits showed implausible values up to 2.5%. The MPA showed the ability to improve the data quality for downstream analyses by detecting implausible observations and to discover physiologically extreme conditions even within complex data structures. At this stage, the MPA is not a fully developed and validated management tool, but rather corresponds to a basic concept for future works, which can be extended and modified as required.


BOLD-fMRI of PTZ-induced seizures in rats.
To develop a non-invasive method for exploring seizure initiation and propagation in the brain of intact experimental animals. We have developed and applied a model-independent statistical method--Hierarchical Cluster Analysis (HCA)--for analyzing BOLD-fMRI data following administration of pentylenetetrazol (PTZ) to intact rats. HCA clusters voxels into groups that share similar time courses and magnitudes of signal change, without any assumptions about when and/or where the seizure begins. Epileptiform spiking activity was monitored by EEG (outside the magnet) following intravenous PTZ (IV-PTZ; n=4) or intraperitoneal PTZ administration (IP-PTZ; n=5). Onset of cortical spiking first occurred at 29+/-16 s (IV-PTZ) and 147+/-29 s (IP-PTZ) following drug delivery. HCA of fMRI data following IV-PTZ (n=4) demonstrated a single dominant cluster, involving the majority of the brain and first activating at 27+/-23s. In contrast, IP-PTZ produced multiple, relatively small, clusters with heterogeneous time courses that varied markedly across animals (n=5); activation of the first cluster (involving cortex) occurred at 130+/-59 s. With both routes of PTZ administration, the timing of the fMRI signal increase correlated with onset of EEG spiking. These experiments demonstrate that fMRI activity associated with seizure activity can be analyzed with a model-independent statistical method. HCA indicated that seizure initiation in the IV- and IP-PTZ models involves multiple regions of sensitivity that vary with route of drug administration and that show significant variability across animal subjects. Even given this heterogeneity, fMRI shows clear differences that are not apparent with typical EEG monitoring procedures, in the activation patterns between IV and IP-PTZ models. These results suggest that fMRI can be used to assess different models and patterns of seizure activation.


Heart failure as interstitial cancer: emergence of a malignant fibroblast phenotype.
A prolonged state of left ventricular pressure overload, commonly caused by hypertension and aortic valve disease, promotes remodelling of the myocardium that can progress to heart failure with preserved ejection fraction (HFpEF). In animal models, a major factor driving progression from pressure-overload hypertrophy (POH) to HFpEF is the activation and proliferation of an abnormal fibroblast phenotype that is resistant to apoptosis, degrades normal stromal matrix and is replaced with a fibrotic matrix structure. A similar fibroblast phenotype has been identified in the stroma of solid cancers. This cancer-associated fibroblast drives tumour growth and invasion. The proliferation and expansion of these abnormal fibroblast populations in both HFpEF and cancer contribute to progression of disease. In early-phase clinical trials, chemotherapeutic agents targeting cancer-associated fibroblasts had antitumour properties. In this Perspectives article, we postulate that, because the abnormal fibroblast populations in POH and cancer have identical characteristics, chemotherapeutic agents targeting the POH-related fibroblast might attenuate the development of myocardial fibrosis, a pathophysiological hallmark of HFpEF. These agents must be designed to target the abnormal fibroblasts with high specificity because many classes of chemotherapeutic drugs can themselves cause myocardial dysfunction and heart failure.


Clinical and prognostic features of spinal meningioma: a thorough analysis from a single neurosurgical center.
The aim of this study was to thoroughly analyze the clinical characteristics of a large cohort of spinal meningioma (SM) from a single neurological center and identify risk factors associated with worse progression free survival and neurological function outcome. Clinical information was retrieved from 483 SM and 9806 cranial meningioma cases who were operated in our center between 2003 and 2013. 194 SM patients who were followed at the main branch were used for prognostic analyses that included both recurrence free survival and neurological functions based on Modified McCormick scale (MMS). Females were predominant (P < 0.001). High grade tumors were not common (WHO grade II, 2.9%; grade III, 1.7%), while the clear cell subtype was frequent within grade II SMs (6/14, 42.9%). Macroscopic total resection was achieved in all SMs (Simpson grade I, 30.9%; grade II, 65.5%; grade III, 3.6%) with a low complications rate (4.6%) and provided neurological improvement in 80 patients (41.2%). Recurrence was seen in 9 cases (4.6%) and associated with high WHO grade, male, prior recurrence, and Simpson grade III. High WHO grade and high Ki-67 index were identified to be independent factors predictive of both neurological function deterioration and impaired post-operative neurological status. Our analysis of the largest SM cohort in scale from a single institution offers a comprehensive view of the clinical characteristics of surgically treated SM, revealing the distinct biology of SM in comparison to its cranial counterparts, and providing guidance to improve surgical management of SM.


Factors affecting length of stay in the pediatric emergency department.
A large volume of visits can cause an emergency department (ED) to become overcrowded, resulting in a longer length of stay (LOS). The objective of this study was to analyze factors affecting the LOS in the pediatric ED. Records of all visits to the pediatric ED of the study hospital, from July 1, 2006 to June 31, 2007, were retrospectively retrieved. Data were collected from the hospital's computerized records system. Eta-squared correlation ratio and Cramer's V test evaluated the associations between variables. Two-thirds of the database was randomized for the classification and regression tree (CART) model-building dataset, and one-third was used for the validation dataset. A total of 29,035 patients visited the pediatric ED during the evaluation period. Of the total visits, 61.1% were due to complaints of fever. The mean LOS was 2.6 ± 4.67 hours, and 74.3% of visits had an LOS of shorter than 2 hours. The CART analysis selected five factors (waiting time for hospitalization, laboratory tests, door-to-physician time, gastrointestinal symptoms, and patient outcome) to produce a total of nine subgroups of patients. The mean LOS of the model-building dataset closely correlated with that of the validation dataset (r(2) = 0.999). Patients who were waiting for hospitalization for less than 8 hours or were not admitted, those without any laboratory tests, those having door-to-physician time less than 60 minutes, and those without any gastrointestinal symptoms had the shortest LOS. Patients who waited for hospitalization for more than 16 hours had the longest LOS.


Method choice in nonfatal self-harm as a predictor of subsequent episodes of self-harm and suicide: implications for clinical practice.
We examined time-varying and time-invariant characteristics of nonfatal intentional self-harm episodes in relation to subsequent episodes of self-harm and suicide. We conducted a follow-up cohort study through 2007 of 3600 patients discharged from hospitals in New Jersey with a primary diagnosis of intentional self-harm in 2003. We determined repetition of self-harm from hospital records and suicide from state registers. Use of methods other than drug overdose and cutting in self-harm events, greater medical severity of nonfatal episodes, and a history of multiple self-harm episodes increased the risk of suicide. However, most suicides occurred without these risk factors. Most suicides took place without intervening episodes of self-harm, and most persons used a low-lethality method (drug overdose or cutting) in their index episode, but switched to a more lethal method in their fatal episode. Our findings suggest that preventing suicide among persons with a history of self-harm must account for the possibility that they will adopt methods with higher case-fatality ratios than they previously tried.


Qingre Huazhuo Jiangsuan Decoction promotes autophagy by inhibiting PI3K/AKT/mTOR signaling pathway to relieve acute gouty arthritis.
Gout belongs to the category of "arthralgia syndrome" in traditional Chinese medicine. It is believed that gout is caused by stagnation of blood stasis, heat, and turbid toxin. Qingre Huazhuo Jiangsuan Decoction (QHJD) is a traditional Chinese medicine prescription developed from the classic Chinese medicine prescription Simiao powder to clear heat, remove turbidity, reduce acid, and reduce inflammation. Now Traditional Chinese Medicine (TCM) physicians often apply it to treat acute gouty arthritis (AGA). However, the mechanism of QHJD in relieving acute gouty arthritis is still unclear, and further research is needed. Here, we aim to explore the potential mechanism of QHJD in relieving acute gouty arthritis. Acute gouty arthritis model was established by injecting monosodium urate (MSU) suspension into knee joint. The pathological state of synovial tissue in each group was evaluated by hematoxylin-eosin (HE) staining. The level of TNF-α, IL-6, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). qRT-PCR was used to detect the mRNA expression of NLRP3, ATG5, ATG7, PI3K, AKT, and mTOR. The protein expression of LC3II/I, p62, ULK1, P-ULK1, Beclin-1, PI3K, AKT, mTOR, P-PI3K, P-AKT, and P-mTOR were detected by Western blot. (1) The level of autophagy protein (mRNA) was significantly up-regulated in QHJD group and rapamycin, while the expression of autophagy protein (mRNA) was significantly downregulated in the 3-methyladenoenoic acid (3 MA) group; (2) QHJD and rapamycin significantly inhibited PI3K/AKT/mTOR pathway, while 3 MA group activated this pathway. (3) It was worth noting that after treatment with QHJD and rapamycin, the inflammatory pathological state of AGA synovial tissue was significantly reduced with the activation of the autophagy gene in knee synovial tissue, and the inhibition of PI3K/AKT/mTOR pathway. This research revealed that QHJD activates autophagy by inhibiting PI3K/AKT/mTOR pathway, thereby relieving acute gouty arthritis.


Abnormal T-cell costimulation by proteins of the extracellular matrix in human renal allografting.
T cells interact with extracellular matrix (ECM) proteins as they pass through the endothelium and move about in various microenvironments. Recent data indicate that such proteins provide strong costimulation to CD3-TCR-complex-mediated T-cell activation. Thus, ECM proteins can regulate T-cell responses via their integrin and non-integrin receptors. We have shown that the costimulating activity of interstitial ECM proteins (collagen type I, fibronectin) is abolished in renal allograft recipients treated with cyclosporin A (CsA) but not azathioprine. In contrast, cyclosporin treatment does not affect potent costimulation provided by the major component of basement membranes, collagen type IV. We suggest that these disturbances in T-cell interactions with ECM proteins caused by CsA therapy may contribute to cyclosporin-induced transplant arteriosclerosis.


A 25-kDa NH2-terminal fragment carries all the antibacterial activities of the human neutrophil 60-kDa bactericidal/permeability-increasing protein.
We have isolated, after limited proteolysis of the bactericidal/permeability-increasing protein (BPI) of human neutrophils, a 25-kDa fragment that possesses the bactericidal and envelope-altering activities of the 60-kDa parent protein. On a molar basis, the fragment is as potent as holo-human BPI against rough Escherichia coli, is more potent than holo-BPI against more resistant smooth E. coli, and retains the specificity of BPI toward Gram-negative bacteria. NH2-terminal amino acid sequence analysis shows that the fragment is derived from the NH2 terminus of the BPI molecule. These findings suggest that all of the molecular determinants of the antibacterial properties of BPI reside within the NH2-terminal 25-kDa segment, implying a novel structural/functional organization for a cytotoxic protein.


Associations between irritable bowel syndrome and non-alcoholic fatty liver disease: A systematic review.
Irritable bowel syndrome (IBS) is associated with obesity and metabolic syndrome. IBS and non-alcoholic fatty liver disease (NAFLD) are highly prevalent entities worldwide and may share similar mechanisms including gut dysbiosis, impaired intestinal mucosal barrier and immune system activation. To systematically review their association according to the Preferred Reporting Items for Systemic Review and Meta-analyses guidelines. PubMed, EMBASE and Cochrane Database of Systematic Reviews were searched for relevant papers. Manual searches were also performed. Six studies were included. Both IBS and NAFLD subjects had significantly more metabolic risk factors like hypertension, obesity, dyslipidaemia and diabetes. Our review showed that 23.2% to 29.4% of NAFLD patients had IBS. IBS was significantly higher in NAFLD patients compared with patients without NAFLD (23.2% vs 12.5%, P < 0.01). A higher proportion of IBS patients had NAFLD (65.8% to 74.0%). IBS patients were three times more likely to have NAFLD compared with non-IBS patients (P < 0.001). Two studies showed a significant correlation between the severity of IBS and NAFLD. The proportion of NAFLD subjects with IBS increased with NAFLD severity. Further prospective studies are warranted to evaluate the relationship and shared pathways between IBS and NAFLD, potentially leading to the development of future therapeutics.


Fate and implication of acetylacetone in photochemical processes for water treatment.
Acetylacetone (AA), due to the peculiar enol-keto structures, has attracted wide scientific interests. In terms of photo-decolorization, it works much more efficiently than the well-known H2O2. However, there is very limited information on the photochemistry of AA in aqueous solutions. Herein, the photolysis kinetics, quantum yield, mass balance, decomposition pathway, and bioavailability of AA during UV irradiation were systematically investigated. It seems that photophysical processes predominated over photochemical ones when AA was irradiated with UV light. Although the quantum yield of AA (0.116) was much lower than that of H2O2 (1.0), the stronger light absorption of AA and the better overlap of the AA absorption spectrum with the solar emission spectrum, as well as the direct energy/electron transfer mechanisms, ensured its high efficiency in photochemical processes. The main degradation products of AA in photochemical processes were similar to the metabolic products in bio-fermentation. Besides, the irradiated AA solution showed a high bioavailability to the cells in activated sludge. Therefore, the UV/AA process might be a promising pre-treatment approach for bio-treatment. The results provide new insights into the photochemical fate and implication of β-diketones in aqueous solutions.


The Role of Pretreatment FDG-PET in Treating Cervical Cancer Patients With Enlarged Pelvic Lymph Node(s) Shown on MRI: A Phase 3 Randomized Trial With Long-Term Follow-Up.
This report is the second analysis of a prospective randomized trial to investigate the impact of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) on cervical cancer patients with enlarged pelvic lymph nodes identified by magnetic resonance imaging. Patients with newly diagnosed cervical cancer with enlarged pelvic lymph nodes but free of enlarged para-aortic lymph nodes (PALN) were eligible. Patients were randomized to receive either pretreatment FDG-PET (PET arm) or not (control arm). The whole pelvis was the standard irradiation field for all patients except those with FDG-avid extrapelvic findings. In all, 129 patients were enrolled. Pretreatment PET detected extrapelvic metastases in 7 patients. No new patient experienced treatment failure during the additional 4-year follow-up period. There were no significant differences between the PET arm and the control arm regarding overall survival, disease-free survival, and freedom from extrapelvic metastasis. In the control arm, 8 of 10 patients with PALN relapse had limited extrapelvic nodal failures; their 5-year disease-specific survival was 34.3%. By contrast, only 1 of 5 patients with PALN relapse in the PET arm experienced such limited failures; their 5-year survival rate was 0%. Although the pretreatment detection of PALN did not translate into survival benefit, it indeed decreased the need for extended-field concurrent chemoradiation therapy.


[A potential contribution of virtual slides and automatic image processing to the estimation of the vascularization of cranial sutures].
Determining age at the time of death is a difficult problem in forensic practice. The study of the vascularization of cranial sutures is an original approach, which may mark the process of synostosis associated with aging. Counting sections of blood vessels on a histological section of cranial suture, raises however a number of practical problems related to the quality of the preparation and to the method of quantification. The purpose of this study is to illustrate the potential contribution of an automatic analysis of virtual slides to overcome these problems. The proposed method of analysis is illustrated from three samples of frontosphenoidal suture whose vessels were immunostained after decalcification of bone tissue. A high resolution image (x 20 objective and microscopic resolution of 0.5 microm) of each microscopic preparation was acquired through a microscopic scanning device. The automatic image analysis protocol takes advantage of a processing of virtual slides at two resolutions. The chosen strategy ensures the identification of the specific area of interest, the enumeration of blood vessel sections on the whole preparation and the visual control of the detected structures. The quantitative estimate of the vascularization of a large structure, such as a cranial suture, can benefit from scanning and fully automatic processing of virtual slides. The automatic analysis requires however an optimal preparation of tissues; decalcification and immunohistochemical staining must be done with great care.


Treatment challenges of persistent ghost cell odontogenic carcinoma: a case report and literature review.
The aim of this case report is to review and compare the clinical, radiologic, histopathologic, and immunohistochemical features, along with the treatment of a case of ghost cell odontogenic carcinoma. In addition, a report of the existing published literature with an emphasis on treatment will be described to provide information on this rare but aggressive tumor. The family of odontogenic ghost cell tumors comprises a spectrum of lesions characterized by odontogenic epithelium with ghost cell keratinization and calcifications. It appears that early detection is vital in proper treatment due to the high possibility of malignant transformation.


Leaf surface specializations of birds and Arthropods in a Panamanian forest.
Most species of Panamanian lowland forest birds specialize on leaf undersurfaces when hunting foliage insects. The few species of leaf surface generalists and leaf upper surface specialists are omnivorous gleaners. We estimate that while over 90% of the avian understory insectivory is directed towards leaf undersurfaces, only 50% of canopy foliage insectivory is directed towards the undersides of leaves. In the low understory we found 70-80% of the arthropods on leaf undersides. The excess use of leaf-bottoms by understory birds may be a result of their greater visibility. It is hypothesized that less proficient insectivores are unable to take advantage of the greater effective density of underleaf insects because they can only efficiently attack the closest leaf surfaces; these closest surfaces will usually be the leaf tops from the branch on which the bird is perched. Alternatively, leaf-top specialists may have special foraging adaptations for overcoming the disadvantages of leaf-top foraging. These adaptations may involve attack behavior (Tachyphonus luctuosus) or searching behavior (Dacnis cayana). Dacnis often used leaf damage as a foraging cue; this may be the first report of a bird using leaf damage for searching for insects. The greater use of leaf upper surfaces by canopy birds may be influenced by four factors: greater seasonality of insects in the canopy favoring omnivores which may be less efficient insectivores; more insects on leaf tops; fewer planar leaf arrangements in canopy plants; or the greater visibility of leaf upper surfaces of the outer shell of foliage of massive trees. Based on the greater number of arthropods on leaf bottoms in the dry season, the higher abundance of smaller insects on leaf bottoms, as well as the greater proportion of insects on leaf tops at cooler higher elevations, we suggest that arthropods prefer leaf bottoms in tropical areas for physiological, not predator avoidance reasons.


Alkyl isocyanates as active site-directed inactivators of guinea pig liver transglutaminase.
Alkyl isocyanates are effective inactivators of guinea pig liver transglutaminase. Based on the specificity of the reaction the protection against inactivation by glutamine substrate, and the essential nature of calcium for the inactivation reaction, it is concluded that these reagents act as amide substrate analogs and, thus function in an active site-specific manner. Support for the contention that inactivation results from alkyl thiocarbamate ester formation through the single active site sulfhydryl group of the enzyme is (a) the loss of one free--SH group and the incorporation of 1 mol of reagent/mol of enzyme in the reaction, (b) similarity in chemical properties of the inactive enzyme derivative formed to those previously reported for another alkyl thiocarbamoylenzyme and an alkyl thiocarbamoylcysteine derivative, and (c) the finding that labeled peptides from digests of [methyl-14C]thiocarbamoyltransglutaminase and those from digests of iodoacetamide-inactivated enzyme occupy similar positions on peptide maps. Transglutaminase was found to be inactivated neither by urethan anlogs of its active ester substrates nor by urea analogs of its amide substrates. It is concluded on the basis of these findings that inactive carbamoylenzyme derivatives are formed only by direct addition of the transglutaminase active--SH group to the isocyanate C--N double bond, and not, like several serine active site enzymes, by nucleophilic displacement with urethan analogs of substrate, or by nucleophilic displacement with urea analogs of substrate.


Cadmium does not inhibit pulsatile prolactin secretion through TRH.
This work was undertaken to analyse the effects of acute cadmium administration on the pulsatile pattern of prolactin release, in adult male rats. For this purpose, animals were cannulated 40 h before the experiment to allow a continuous blood withdrawal. Two hours after the administration of one dose of cadmium chloride (4.5 mg kg1), the pulsatile pattern of prolactin, during three hours, was studied. The effects of two pulses of thyrotropin-releasing hormone (TRH) (1 microgram per rat), given 60 and 120 min after starting the period of blood sampling, were studied. The mean values of prolactin during the bleeding period and the absolute pulse amplitude were decreased by acute cadmium chloride administration. However, no changes in any other parameters of prolactin pulsatility were observed. TRH administration to control rats increased mean prolactin levels, and absolute and relative pulse amplitudes, but decreased the mean half-life of the hormone. In animals pretreated with cadmium, TRH increased the mean levels of prolactin, and absolute and relative amplitudes of the hormone pulses. No other parameter studied was changed by TRH in cadmium pretreated rats. These data suggest that acute administration of cadmium did not inhibit the pulsatile prolactin release through TRH.


Effect of increased intake of concentrates and sodium butyrate supplementation on ruminal epithelium structure and function in growing rams.
Increased ruminal butyrate production is considered to have a positive impact on rumen epithelium growth and function. However, excessive ruminal butyrate production may affect the rumen negatively, particularly when the rumen is already challenged with low pH. The aim of this study was to determine the effect of the inclusion of concentrates in the diet and sodium butyrate (SB) supplementation on ruminal epithelium growth and function in growing rams. Forty-two rams (27.8 ± 7.3 kg; 9-14 months of age) were allocated into six treatments and fed a diet with low (22.5% of diet DM; LOW) or high (60% of diet DM; HIGH) inclusion of concentrates in combination with no (SB0), 1.6% (SB1.6) or 3.2% (SB3.2) of diet DM inclusion of SB. There was no impact of the investigated factors on papilla dimensions and mucosa surface area, either in the atrium ruminis or ventral rumen (P ≥ 0.11). Stratum corneum thickness was higher for HIGH compared to LOW treatments (P ≤ 0.04), independently of the location in the rumen. In the atrium ruminis, the epithelium and living strata thickness quadratically increased due to SB supplementation for LOW treatments but quadratically decreased for HIGH treatments (concentrate inclusion × butyrate supplementation interaction; P ≤ 0.03); conversely, in the ventral sac of the rumen, a thicker epithelium was observed due to both increased concentrate inclusion in the diet and SB supplementation (P < 0.01) but living strata thickness was increased only by SB supplementation (linear effect; P < 0.01). The epithelium damage index in the ventral sac of the rumen was higher for LOW compared to HIGH treatments (P = 0.02). Increased inclusion of concentrates in the diet increased mRNA expression of monocarboxylate transporter 1 in both the epithelium of the atrium ruminis and ventral rumen, occludin in the epithelium of the atrium ruminis and downregulated in adenoma in the epithelium of the ventral rumen (P ≤ 0.02). Protein expression of claudin-4 in the epithelium of the ventral rumen was the highest for the HIGH/SB1.6 and HIGH/SB3.2 treatments (significant effect of interaction between main effects; P < 0.01). Under the conditions of the current study, increased intake of concentrates had mostly positive effects on ruminal epithelium in growing rams, and the same was observed for the effect of SB supplementation. However, the effect of SB supplementation was at least partially affected by the inclusion of concentrates in the diet.


Combined metabolome and transcriptome analysis reveal the mechanism of selenate influence on the growth and quality of cabbage (Brassica oleracea var. capitata L.).
Selenium is an essential trace element for human and animal health, and an appropriate amount of Se can promote the growth and development of plants. Cabbage is a popular cruciferous vegetable with a good ability to accumulate Se, and Se-enriched cabbage can be used as an important Se source for humans. However, the effects of Se-enriched cultivation and the Se accumulation mechanism in cabbage are still unclear. In this study, the effects of different concentrations (0, 0.1, 0.2, 0.4, 0.8, and 1.6 mmol/L) of selenate on cabbage growth and quality were explored. A low concentration of selenate (0.1 mmol/L) promoted growth and nutritional quality. The contents of total Se, S, selenocystine, and selenomethionine significantly increased following selenate application. Important secondary metabolites, namely glucosinolates, phenolic acids, and flavonoids, participate in the response to selenate in cabbage. Comparative transcriptome and metabolomics analysis revealed that SULTR2.2, SULTR3.1, APS, APK2, HMT, MMT, and NTR2 played important roles in Se absorption and conversion. Additionally, the SUR1, UGT74B1, and ST5b genes and cytochrome P450 family genes CYP83A1, CYP79A2, and CYP79F1 may be the crucial genes in the glucosinolates biosynthesis and regulation pathway. The PAL, 4CL, CAD, CHS3, FLS, and CYP73A5 genes were involved in flavonoid and phenolic acid accumulation under selenate treatment. These results reveal the internal relationships in the regulatory network of Se metabolism and secondary metabolite biosynthesis in cabbage and help further the understanding of the physiological and molecular mechanism of how Se biofortification affects cabbage quality, thereby providing genetic resources and a technical basis for the breeding and cultivation of Se-enriched cabbage with excellent nutritional quality.


NO-sensitive guanylyl cyclase discriminates pericyte-derived interstitial from intra-alveolar myofibroblasts in murine pulmonary fibrosis.
The origin of αSMA-positive myofibroblasts, key players within organ fibrosis, is still not fully elucidated. Pericytes have been discussed as myofibroblast progenitors in several organs including the lung. Using tamoxifen-inducible PDGFRβ-tdTomato mice (PDGFRβ-CreERT2; R26tdTomato) lineage of lung pericytes was traced. To induce lung fibrosis, a single orotracheal dose of bleomycin was given. Lung tissue was investigated by immunofluorescence analyses, hydroxyproline collagen assay and RT-qPCR. Lineage tracing combined with immunofluorescence for nitric oxide-sensitive guanylyl cyclase (NO-GC) as marker for PDGFRβ-positive pericytes allows differentiating two types of αSMA-expressing myofibroblasts in murine pulmonary fibrosis: (1) interstitial myofibroblasts that localize in the alveolar wall, derive from PDGFRβ+ pericytes, express NO-GC and produce collagen 1. (2) intra-alveolar myofibroblasts which do not derive from pericytes (but express PDGFRβ de novo after injury), are negative for NO-GC, have a large multipolar shape and appear to spread over several alveoli within the injured areas. Moreover, NO-GC expression is reduced during fibrosis, i.e., after pericyte-to-myofibroblast transition. In summary, αSMA/PDGFRβ-positive myofibroblasts should not be addressed as a homogeneous target cell type within pulmonary fibrosis.


Determination of diosmin in pharmaceutical formulations using Fourier transform infrared spectrophotometry.
A Fourier transform infrared (FT-IR) spectrometric method was developed for the rapid, direct measurement of diosmin in different pharmaceutical drugs. Conventional KBr-spectra were compared for best determination of active substance in commercial preparations. The Beer-Lambert law and two chemometric approaches, partial least squares (PLS) and principal component regression (PCR+) methods, were tried in data processing.


Development of a novel PTD-mediated IVT-mRNA delivery platform for potential protein replacement therapy of metabolic/genetic disorders.
The potential clinical applications of the powerful in vitro-transcribed (IVT)-mRNAs, to restore defective protein functions, strongly depend on their successful intracellular delivery and transient translation through the development of safe and efficient delivery platforms. In this study, an innovative (international patent-pending) methodology was developed, combining the IVT-mRNAs with the protein transduction domain (PTD) technology, as an efficient delivery platform. Based on the PTD technology, which enables the intracellular delivery of various cargoes intracellularly, successful conjugation of a PTD to the IVT-mRNAs was achieved and evaluated by band-shift assay and NMR spectroscopy. In addition, the PTD-IVT-mRNAs were applied and evaluated in two protein-disease models, including the mitochondrial disorder fatal infantile cardioencephalomyopathy and cytochrome c oxidase (COX) deficiency (attributed to SCO2 gene mutations) and β-thalassemia. The PTD-IVT-mRNA of SCO2 was successfully transduced and translated to the corresponding Sco2 protein inside the primary fibroblasts of a SCO2/COX-deficient patient, whereas the PTD-IVT-mRNA of β-globin was transduced and translated in bone marrow cells, derived from three β-thalassemic patients. The transducibility and the structural stability of the PDT-IVT-mRNAs, in both cases, were confirmed at the RNA and protein levels. We propose that our novel delivery platform could be clinically applicable as a protein therapy for metabolic/genetic disorders.


G9a inhibition promotes neuronal differentiation of human bone marrow mesenchymal stem cells through the transcriptional induction of RE-1 containing neuronal specific genes.
Recent studies have shown that epigenomic modifications are significantly associated with neuronal differentiation. Many neuronal specific genes contain the repressor element-1 (RE-1), which recruits epigenetic modulators, such as the histone methyltransferase G9a and interrupts the expression of neuronal genes in non-neuronal cells. This study investigated the functional role of G9a during neuronal differentiation of human bone marrow mesenchymal stem cells (BM-MSCs). Human BM-MSCs treated with the G9a inhibitor BIX01294 showed an increased expression of various neuronal-lineage genes. Using genomic sequence analysis, we identified RE-1 consensus sequences in the proximal region of several neuronal-specific genes. Chromatin immunoprecipitation (ChIP) assay results have showed that H3K9me2 (dimethylation of lysine 9 on histone 3) occupancy at RE-1-containing sequences from neuronal-specific genes was significantly decreased in BIX01294-MSCs. When BIX01294-MSCs were differentiated with neuronal induction medium, cells differentiated more effectively into neuron-like cells, complete with a cell body and dendrites. Expression of neuronal-specific genes containing the RE-1 sequences was significantly increased in differentiated BIX01294-MSCs, as confirmed by immunocytochemical staining and immunoblotting. Thus, this study shows that BIX01294 pretreated human BM-MSCs can be effectively differentiated into neuron-like cells by induced expression of neuronal-specific genes containing RE-1 sequences.


High adherence to all-oral directly acting antiviral HCV therapy among an inner-city patient population in a phase 2a study.
As treatment for chronic hepatitis C (HCV) virus has evolved to all-oral, interferon-free directly acting antiviral (DAA) therapy, the impact of these improvements on patient adherence has not been described. Medication adherence was measured in 60 HCV, genotype-1, treatment-naïve participants enrolled in a phase 2a clinical trial at the National Institutes of Health and community clinics. Participants received either ledipasvir/sofosbuvir (LDV/SOF) (90 mg/400 mg) (one pill) daily for 12 weeks, LDV/SOF + GS-9451 (80 mg/day) (two pills) daily for 6 weeks, or LDV/SOF + GS-9669 (500 mg twice daily; three pills, two in the morning, one in the evening) for 6 weeks. Adherence was measured using medication event monitoring system (MEMS) caps, pill counts and patient report. Overall adherence to DAAs was high. Adherence declined over the course of the 12-week treatment (p = 0.04). While controlled psychiatric disease or symptoms of depression did not influence adherence, recent drug use was a risk factor for non-adherence to 12-week (p = 0.01), but not 6-week regimens. Adherence as measured by MEMS was lower than by patient report. Adherence to short courses of DAA therapy with 1-3 pills a day was excellent in an urban population with multiple risk factors for non-adherence.


Capacity-Achieving Input Distributions of Additive Vector Gaussian Noise Channels: Even-Moment Constraints and Unbounded or Compact Support.
We investigate the support of a capacity-achieving input to a vector-valued Gaussian noise channel. The input is subjected to a radial even-moment constraint and is either allowed to take any value in Rn or is restricted to a given compact subset of Rn. It is shown that the support of the capacity-achieving distribution is composed of a countable union of submanifolds, each with a dimension of n-1 or less. When the input is restricted to a compact subset of Rn, this union is finite. Finally, the support of the capacity-achieving distribution is shown to have Lebesgue measure 0 and to be nowhere dense in Rn.


Quality of life profile--adolescent version: assessing the relationship of covariates to scale scores using structural equation modeling.
This study investigates the effects of covariates on adolescent quality of life (QOL) using structural equation modeling (SEM). The Quality of Life Profile--Adolescent Version, a generic 54 item self-reported questionnaire, was administered to 363 adolescents aged between 10 and 18 years who were enrolled in 20 secondary schools within metropolitan Perth, the capital of Western Australia. Previous results based on a second-order SEM provided preliminary support for measuring adolescent QOL in terms of physical health, psychological, social, environment and opportunities for growth and development. The second-order model is further enhanced in this study by incorporating covariates age, presence of a chronic condition, amount of control and perception of health. The amount of control the adolescent perceived had a significant positive impact on QOL, whereas poorer reports of health had a negative impact. However, the presence of a chronic condition was found to have little effect on QOL, while the modest inverse relationship between age and QOL was not statistically significant.