Evaluation of a Raman Chemometric Method for Detecting Protein Structural Conformational Changes in Solution.
Raman scattering shows promise as a powerful routine tool, to determine both secondary and the smaller tertiary structural changes that precede aggregation in both solutions and solids. A method was developed utilizing principal component analysis (PCA) of Raman spectra for detection of small, but meaningful, pH induced changes in tertiary protein structure linked to aggregate formation using α-lactalbumin solutions as a model. The sample preparation and spectral parameters, were optimized for a bulk Raman probe. Analysis of large regions (600-1850 cm-1) yielded principal component (PC) scores useful for semi-quantitative comparison of protein conformation between formulations. PC loadings corresponded to specific structural peaks known to change with solution pH. PCA of circular dichroism (CD) spectra of dilute solutions yielded similar results. Sucrose is a common formulation excipient with a Raman spectrum that overlaps many protein peaks. With sucrose in the protein solution, the ability of PCA to discern protein structural changes from the Raman spectra was somewhat reduced. Analysis of a more limited spectral region (1530-1780 cm-1) with negligible sucrose spectral contribution improved the discrimination of protein conformational states. The new Raman method accurately distinguished differences in protein structure in concentrated solutions. The long-term goal is to explore Raman characterization as a routine monitoring tool of protein stability in both solution and solid states.


Alteration of the conformation of proteins in red blood cell membranes and in solution by fixatives used in electron microscopy.
The effects of several commonly employed fixatives on the three-dimensional conformations of two soluble proteins and the protein of intact red blood cell membranes have been studied by means of circular dichroism measurements in the spectral region of the peptide absorption bands. The fixatives used produced significant and parallel conformational changes in all of the proteins, in the increasing order: glutaraldehyde; OsO(4); glutaraldehyde followed by OsO(4); and KMnO(4). The last two treatments obliterated most of the helical character of the proteins. The significance of these observations to the preparation of specimens for electron microscopy is discussed.


[Solitary intraspinal dorsal neurinoma. A case report].
Most spiral neurinomas are intradural extramedullary tumours. Intramedullary neurinomas are rare: only 20 cases have been published. The authors report a new case of solitary intramedullary dorsal neurinoma and review the main diagnostic and therapeutic problems raised by this type of tumour.


Primary transpupillary thermotherapy for choroidal melanoma in 391 cases: importance of risk factors in tumor control.
To report the long-term outcome of primary transpupillary thermotherapy (TTT) for choroidal melanoma. Retrospective review of medical records. We included 391 patients with choroidal melanoma treated between 1995 and 2012 at the Oncology Service, Wills Eye Hospital, Philadelphia. We delivered TTT with an infrared diode laser. Local tumor recurrence, Snellen visual acuity after TTT, and distant metastasis. Of 391 patients, 311 (80%) were treated from 1995 to 2000 and 80 (20%) from 2001 to 2012. Tumors in the 2001 to 2012 group were ultrasonographically thinner (2.2 vs. 2.7 mm), more distant from the optic disc (3.2 vs. 2.5 mm) and foveola (4.0 vs. 2.0 mm), were less often located in the macular area (14% vs. 40%), and had lower rates of acoustic hollowness on B-scan ultrasonography (63% vs. 84%), subretinal fluid (58% vs. 90%), and orange pigment (50% vs. 70%). Kaplan-Meier estimates for tumor recurrence in the 1995 to 2000 group were 29% at 5 years and 42% at 10 years, whereas estimates for tumor recurrence in the 2001-2012 group were 11% at 5 years and 15% at 10 years. Of 108 recurrent tumors 20 were controlled with additional TTT and 62 required plaque radiation (n=60) or proton beam radiation (n=2), with enucleation necessary in 26 patients. Tumor recurrence correlated with the number of high-risk tumor features: 10-year recurrence was 18% in those with 1 or 2 risk factors, 35% in those with 3 to 5 factors, and 55% in those with 6 or 7 factors. On multivariate analysis, features predictive of tumor recurrence were presence of symptoms (P<0.001), shorter distance between the tumor and the optic disc (P=0.026), subretinal fluid (P=0.035), thickness of residual tumor scar (P<0.001), and elevation of residual tumor scar (P<0.001). The only factor predictive of extraocular tumor extension was intraocular tumor recurrence after TTT treated with additional TTT (P=0.007). Presence of orange pigment before TTT (P=0.019), tumor recurrence (P=0.002), and extraocular tumor extension (P=0.017) were predictive of distant metastasis. This study shows a direct correlation between a larger number of high-risk tumor features and higher rates of tumor recurrence after primary TTT of (small) choroidal melanoma. We advise that, when possible, small choroidal melanomas with multiple risk factors be treated with methods other than TTT.


Long non-coding RNA DARS-AS1 promotes tumor progression by directly suppressing PACT-mediated cellular stress.
Cancer cells evolve various mechanisms to overcome cellular stresses and maintain progression. Protein kinase R (PKR) and its protein activator (PACT) are the initial responders in monitoring diverse stress signals and lead to inhibition of cell proliferation and cell apoptosis in consequence. However, the regulation of PACT-PKR pathway in cancer cells remains largely unknown. Herein, we identify that the long non-coding RNA (lncRNA) aspartyl-tRNA synthetase antisense RNA 1 (DARS-AS1) is directly involved in the inhibition of the PACT-PKR pathway and promotes the proliferation of cancer cells. Using large-scale CRISPRi functional screening of 971 cancer-associated lncRNAs, we find that DARS-AS1 is associated with significantly enhanced proliferation of cancer cells. Accordingly, knocking down DARS-AS1 inhibits cell proliferation of multiple cancer cell lines and promotes cancer cell apoptosis in vitro and significantly reduces tumor growth in vivo. Mechanistically, DARS-AS1 directly binds to the activator domain of PACT and prevents PACT-PKR interaction, thereby decreasing PKR activation, eIF2α phosphorylation and inhibiting apoptotic cell death. Clinically, DARS-AS1 is broadly expressed across multiple cancers and the increased expression of this lncRNA indicates poor prognosis. This study elucidates the lncRNA DARS-AS1 directed cancer-specific modulation of the PACT-PKR pathway and provides another target for cancer prognosis and therapeutic treatment.


Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM.
Light sheet fluorescence microscopy (LSFM) is rapidly becoming an essential technology for mesoscopic imaging of samples such as embryos and adult mouse organs. However, LSFM can suffer from optical artifacts for which there is no intrinsic solution. The attenuation of light due to absorbing material causes "shadow" artifacts along both the illumination and detection paths. Several approaches have been introduced to reduce this problem, including scanning illumination and multi-view imaging. However, neither of these approaches completely eliminates the problem. If the distribution of the absorbing material is complex, shadows cannot be avoided. We introduce a new approach that relies on multi-modal integration of two very different mesoscopic techniques. Unlike LSFM, optical projection tomography (OPT) can operate in transmission mode to create a voxel map of the 3D distribution of the sample's optical attenuation. Here, we demonstrate a hybrid instrument (OPTiSPIM) that can quantify this attenuation and use the information to correct the shadow artifacts of LSFM.


Defective interfering particles of VSVNJ (Ogden), generated by heat treatment, contain multiple internal genomic deletions.
Defective interfering (DI) particles have been isolated from a heat-resistant strain of the New Jersey (Ogden) serotype of vesicular stomatitis virus (VSV). Most of these DI particles contain various portions of all five cistrons of VSV. The two largest DI particles, NJ-121 and NJ-PG2, represent approximately 60% of the standard virus genome and contain both the positive and negative strand leader RNA templates. These two DI particles are transcriptionally active and synthesize both the positive and negative strand leader RNAs in vitro. Virion RNA probe-mRNA hybridizations and cDNA probe-virion RNA hybridizations have shown that NJ-121 contains mainly sequences from the L and G genes. In contrast, NJ-PG2 has portions of the sequences from all five genes of VSV. Smaller DI particles, NJ-121a, NJ-121b, NJ-PG1, and NJ-JM2 representing approximately 50, 38, 28, and 25% of the standard virus genome respectively, were also generated. These DI particles did not have sequences complementary to the positive strand leader RNA template. The mRNA hybridization patterns and results of the genomic RNAs hybridizing to cDNAs of N, NS, M, and G genes of these DI particles showed that they contain parts of information from all five cistrons. Most of the DI particles appear to be generated by multiple deletions throughout the standard virus genome. None of these DI particles interfered heterotypically with VSVIND-HR in BHK21, R(B77), or L2 cells. However, they interfered well with infection by VSVNJ (Hazelhurst).


Clinical trial readiness study of distal myopathy and dysphagia in nephropathic cystinosis.
Nephropathic cystinosis is a lysosomal storage disorder with late-onset systemic complications, such as myopathy and dysphagia. Currently employed outcome measures lack sensitivity and responsiveness for dysphagia and myopathy, a limitation to clinical trial readiness. We evaluated 20 patients with nephropathic cystinosis in two visits over the course of a year to identify outcomes sensitive to detect changes over time. Patients also underwent an expiratory muscle strength training program to assess any effects on aspiration and dysphagia. There were significant differences in the Timed Up and Go Test (TUG) and Timed 25-Foot Walk (25-FW) between baseline and 1-y follow-up (P < .05). Maximum expiratory pressure (MEP) and peak cough flow (PCF) significantly improved following respiratory training (P < .05). Improved respiratory outcomes may enhance patients ability to expel aspirated material from the airway, stave off pulmonary sequelae associated with chronic aspiration, and yield an overall improvement in physical health and well-being.


Bioinformatically-predicted varicella zoster virus small non-coding RNAs are expressed in lytically-infected epithelial cells and neurons.
Most herpesviruses use both host and viral small non-coding RNAs (sncRNA), especially microRNA, to modulate infection. Bioinformatic analyses of NGS data obtained from Varicella Zoster virus (VZV)-infected cells predicted 24 VZVsncRNA, seven of which were confirmed to be expressed in infected fibroblasts and neurons using stem-loop quantitative reverse transcription PCR (SL-PCR). We here assayed for the expression of all 24 of the bioinformatically predicted VZVsncRNA in cells productively infected by VZV using SL-PCR. 23 of the 24 predicted sequences were detected in VZV-infected ARPE19 cells and 19 of the 24 sequences in infected human neurons generated by two methods from embryonic stem cells. We also show that blocking one of two newly-tested VZV-encoded sncRNA using locked nucleotide antagonists significantly increased viral replication. These findings suggest that further study of VZV encoded sncRNA could elucidate an additional level of regulation of the life cycle of this pathogenic human herpesvirus.


Identification of two urinary metabolites of rats treated with acrylonitrile; influence of several inhibitors on the mutagenicity of those urines.
Urines collected from rats injected with acrylonitrile (ACN) were mutagenic towards Salmonella typhimurium TA1530; this activity was reduced when the animals were pretreated by pyrazole (inhibitor of alcohol dehydrogenase) and suppressed after pretreatment either by CoCl2 and SKF 525-A (inhibitors of the mixed-function oxidases system) or by trichloroacetonitrile (radical trapping agent). On the other hand, two urinary metabolites (cyanoethanol and cyanoacetic acid) have been detected by gas chromatography. One possible scheme for the in vivo metabolism of ACN is presented which postulates the intermediate formation of a radical species and of an epoxide.


Transdermal self-permeation enhancement of ibuprofen.
The objective of this study was to prepare saturated solutions of ibuprofen, of different concentrations, and to investigate their effect on permeation of ibuprofen across rat epidermis. Ibuprofen saturated solutions were prepared using 0.1, 0.2, 0.3 and 0.4 M disodium hydrogen phosphate solution (DHP). The solubility of ibuprofen in DHP increased as the molarity of DHP increased. Thus the four saturated solutions of ibuprofen (0.1M-DHP-IBU, 0.2M-DHP-IBU, 0.3M-DHP-IBU and 0.4M-DHP-IBU) have different concentrations of the same drug, and showed same pH (pH 7.0+/-1). The permeability study was also carried out using human epidermis and silastic membrane. Permeation rate of ibuprofen across rat epidermis and human epidermis from 0.4M-DHP-IBU was much greater than from 0.1M-DHP-IBU. The magnitudes of increase in the drug flux were 46.4-fold with rat epidermis and 9.4-fold with human epidermis. Such a great increase in drug flux was not observed with silastic membrane, only 1.4-fold. This suggests that the increased drug flux is likely due to drug-skin interaction and not the increased concentration of ibuprofen per se. Surface tension (ST) measurements of DHP versus ibuprofen concentration showed ST reduction of DHP, from 72 to 27.9 dyn/cm. This is an indication that ibuprofen acted as ionic surfactant and the observed skin permeability enhancement is attributed to disruption of stratum corneum barrier. Results of DSC study supported this assumption. DSC of untreated rat stratum corneum samples showed lipid transitions at 41.9+/-0.0 degrees C (T1), 55.1+/-1.6 degrees C (T(x)), 70.2+/-0.1 degrees C (T2) and 77.5+/-0.1 degrees C (T3), while those pretreated with 0.4M-DHP-IBU did not show the first three lipid transitions. Also, pretreatment of rat epidermis with 0.4M-DHP-IBU enhanced permeation of diclofenac sodium greater than 1250-fold. This corroborates that ibuprofen not only enhances its own permeation but also that of other drugs, such as diclofenac sodium.


Fat Grafting as Regenerative Surgery: A Current Review.
Autologous fat grafting has long been regarded as an ideal filler, lauded for its ability to restore soft-tissue contour. In recent times, fat grafting has exhibited regenerative capacity, largely secondary to the action of adipose-derived stem cells and preadipocytes in the stromal vascular fraction of adipose tissue. This has birthed a new field of regenerative surgery with benefits germane to scarring, fibrosis, atrophy, burns, neuropathic pain, and autoimmune disease. The broad clinical applications of regenerative fat grafting have the potential to improve quality of life through functional and aesthetic improvement. Fat grafting has the potential to serve as a regenerative option for difficult clinical problems that cannot be treated effectively at the present time. Fat grafting also exhibits angiogenic and immunomodulatory properties in the context of autoimmune disease. The broad clinical applications of regenerative fat grafting have the potential to improve quality of life both functionally and aesthetically. It may present a less invasive avenue for clinical issues that today necessitate conventional surgical techniques. However, regenerative fat grafting is still in its infancy; further research is required to ascertain evidence-based protocols for the various clinical indications and better understand the precise regenerative mechanisms after fat grafting.


ANN application for prediction of atmospheric nitrogen deposition to aquatic ecosystems.
The occurrences of increased atmospheric nitrogen deposition (ADN) in Southeast Asia during smoke haze episodes have undesired consequences on receiving aquatic ecosystems. A successful prediction of episodic ADN will allow a quantitative understanding of its possible impacts. In this study, an artificial neural network (ANN) model is used to estimate atmospheric deposition of total nitrogen (TN) and organic nitrogen (ON) concentrations to coastal aquatic ecosystems. The selected model input variables were nitrogen species from atmospheric deposition, Total Suspended Particulates, Pollutant Standards Index and meteorological parameters. ANN models predictions were also compared with multiple linear regression model having the same inputs and output. ANN model performance was found relatively more accurate in its predictions and adequate even for high-concentration events with acceptable minimum error. The developed ANN model can be used as a forecasting tool to complement the current TN and ON analysis within the atmospheric deposition-monitoring program in the region.


Federal demonstration funds at the interface of mental health and criminal justice.
A jail diversion program was implemented for offenders with co-occurring mental health and substance abuse disorders (COD) in Lane County, Oregon. This paper discusses whether the program resulted in any sustained community service systems change. Interviewees were asked questions about any systems-level changes that interviewees were aware of in corrections, criminal justice, or treatment where offenders with COD were involved. Results showed evidence of system-level changes in corrections, criminal justice, and treatment that were attributed to various aspects of our "boundary spanning" jail diversion approach with the COD population. The researchers also interpreted the results as evidence of the efficacy of using research to promote change.


Vesicle Adhesion and Fusion Studied by Small-Angle X-Ray Scattering.
We have studied the adhesion state (also denoted by docking state) of lipid vesicles as induced by the divalent ions Ca2+ or Mg2+ at well-controlled ion concentration, lipid composition, and charge density. The bilayer structure and the interbilayer distance in the docking state were analyzed by small-angle x-ray scattering. A strong adhesion state was observed for DOPC:DOPS vesicles, indicating like-charge attraction resulting from ion correlations. The observed interbilayer separations of ∼1.6 nm agree quantitatively with the predictions of electrostatics in the strong coupling regime. Although this phenomenon was observed when mixing anionic and zwitterionic (or neutral) lipids, pure anionic membranes (DOPS) with highest charge density σ resulted in a direct phase transition to a multilamellar state, which must be accompanied by rupture and fusion of vesicles. To extend the structural assay toward protein-controlled docking and fusion, we have characterized reconstituted N-ethylmaleimide-sensitive factor attachment protein receptors in controlled proteoliposome suspensions by small-angle x-ray scattering.


Parent-of-origin detection and chromosome-scale haplotyping using long-read DNA methylation sequencing and Strand-seq.
Hundreds of loci in human genomes have alleles that are methylated differentially according to their parent of origin. These imprinted loci generally show little variation across tissues, individuals, and populations. We show that such loci can be used to distinguish the maternal and paternal homologs for all human autosomes without the need for the parental DNA. We integrate methylation-detecting nanopore sequencing with the long-range phase information in Strand-seq data to determine the parent of origin of chromosome-length haplotypes for both DNA sequence and DNA methylation in five trios with diverse genetic backgrounds. The parent of origin was correctly inferred for all autosomes with an average mismatch error rate of 0.31% for SNVs and 1.89% for insertions or deletions (indels). Because our method can determine whether an inherited disease allele originated from the mother or the father, we predict that it will improve the diagnosis and management of many genetic diseases.


Assessment of Pain in Glaucoma Patients Undergoing Micropulse Transscleral Laser Therapy.
This study aimed to assess the pain experienced during micropulse transscleral laser therapy (MPTLT) and overnight thereafter and explore the factors associated with the pain. This prospective study included 100 eyes of 81 glaucoma patients undergoing MPTLT under retrobulbar anesthesia. All patients were asked to rate both types of pain using a numerical rating scale (NRS). The risk factors were explored using multivariable mixed-effects ordinal logistic regression. The mean (SD) NRS pain score during the procedure was 3.57 (3.41) (range 0-10), which included no, mild, moderate, and severe pain in 30 (30%), 33 (33%), 17 (17%), and 20 (20%) eyes, respectively. The mean (SD) NRS score of overnight pain was 2.99 (2.28) (range 0-9), which included no, mild, moderate, and severe pain in 17 (17%), 59 (59%), 17 (17%), and 7 (7%) eyes, respectively. Twenty-seven (27%) eyes reported worse pain overnight than during the procedure. Increased age, initial intraocular pressure, and pain during the procedure were significantly associated with increased overnight pain (p < 0.05). Up to a fourth of eyes had worse pain after discharge. Older age, initial intraocular pressure, and pain during the procedure were risk factors for higher levels of overnight pain.


Increased NF-kB activity in osteoprogenitor-lineage cells impairs the balance of bone versus fat in the marrow of skeletally mature mice.
"Senile osteoporosis" is defined as significant aging-associated bone loss, and is accompanied by increased fat in the bone marrow. The proportion of adipocytes in bone marrow is inversely correlated with bone formation, and is associated with increased risk of fracture. NF-κB is a transcription factor that functions as a master regulator of inflammation and bone remodeling. NF-κB activity increases during aging; furthermore, constitutive activation of NF-κB significantly impairs skeletal development in neonatal mice. However, the effects of NF-κB activation using a skeletally mature animal model have not been examined. In the current study, an osteoprogenitor (OP)-specific, doxycycline-regulated NF-κB activated transgenic mouse model (iNF-κB/OP) was generated to investigate the role of NF-κB in bone remodeling in skeletally mature mice. Reduced osteogenesis in the OP-lineage cells isolated from iNF-κB/OP mice was only observed in the absence of doxycycline in vitro. Bone mineral density in the metaphyseal regions of femurs and tibias was reduced in iNF-κB/OP mice. No significant differences in bone volume fraction and cortical bone thickness were observed. Osmium-stained bone marrow fat was increased in epiphyseal and metaphyseal areas in the tibias of iNF-κB/OP mice. These findings suggest that targeting NF-κB activity as a therapeutic strategy may improve bone healing and prevent aging-associated bone loss in aged patients.


Strategies for managing the destruction of calcar femorale.
The calcar femorale was identified long ago. However, our current understanding of the calcar is insufficient, and its related concepts are sometimes confused. The calcar femoral is an important anatomical structure of the proximal femur, and its function can be overlooked. In trauma, tumors, or other diseases, the calcar femorale can be destroyed or changed pathologically. As a result, the mechanical structure of the proximal femur becomes destroyed, causing pathological fractures. How to address the destruction of the calcar femorale or the damage to the calcar femorale is discussed in this article. Destruction of the calcar femorale is accompanied by many conditions, including trauma, tumors, and other diseases. The types of hip fractures caused by trauma include femoral neck fractures and intertrochanteric fractures. Dynamic hip screws, proximal femoral nail anti-rotation, and multiple parallel cannulate pins can be used in different conditions. When metastatic and primary bone tumors involve the calcar femorale, endoprostheses are widely used. Other diseases, such as fibrous dysplasia and aneurysmal bone cyst are treated differently. The calcar femorale can redistribute stresses and the destruction of the calcar femorale can lead to an increase in posterior medial stress. Many factors need to be considered when deciding whether to reconstruct the calcar femorale. Effective treatment strategies for managing the destruction of calcar femorale will need first establishing the precise mechanism of the destruction of the calcar and then designing therapies towards these mechanisms. Further investigation to the calcar needs to be carried out.


N,N'-Diphenyl-but-2-enediamide.
In the title compound, C(16)H(14)N(2)O(2), the conformations of the N-H and C=O bonds in the C-NH-CO-CH =CH-CO-NH-C segment are anti to each other. The two C=O bonds are also anti to each other. The two phenyl rings make an inter-planar angle of 41.2 (1)°. An intra-molecular N-H⋯O hydrogen bond occurs. In the crystal, inter-molecular N-H⋯O hydrogen bonding links the mol-ecules into infinite chains along the a axis.


Modulation of central thalamic oscillations during emotional-cognitive processing in chronic disorder of consciousness.
We report on thalamic recordings in a patient with chronic disorder of consciousness (DOC). Implantation of central thalamic deep brain stimulation (CT-DBS) electrodes was chosen, as this treatment has been reported to display beneficial effects with respect to behavioural responsiveness in DOC. Local field potential (LFP) oscillations were recorded from central thalamic electrodes and their changes elicited by speech stimuli consisting either of familiar voices addressing the patient or unfamiliar non-addressing phrases were studied. In response to familiar-addressing speech we observed modulation of oscillatory activity in the beta and theta band within the central thalamus accompanied by an increase in thalamocortical coherence in the theta band. Furthermore, the theta phase was coupled to the amplitude of gamma locally in the thalamus. These findings indicate a local and long-range cross-frequency response which is not only indicative of the principle involvement of the central thalamus in processing emotional and cognitive information, but also point towards intact physiological functions that may serve as a marker in diagnosing DOC patients and determining novel targets and parameters concerning therapeutic efforts.


Prevalence and impact of antimicrobial resistance in gastrointestinal infections: A review.
Antimicrobial resistance has become a worldwide problem due to its excessive increase in recent years. The aim of the present review was to bring together data from different articles describing the levels of antimicrobial resistance in the most common gastrointestinal infections reported across the globe. The literature search was carried out in Google Scholar, Medline, Embase, and Pubmed, with the terms "antimicrobial resistance", "resistance in gastrointestinal disorders", and "resistance in amoebiasis", in Spanish and English. Mexican treatment guidelines and consensuses from 2017 to the present were utilized. Publications from the last ten years were chosen to describe the level of resistance. They had adequate sample sizes, the Material and Methods sections were precise, and they included multicenter studies, national and international consensuses, meta-analyses, systematic reviews, and extensive texts. The final number of articles was 51. The microorganisms that demonstrated the highest percentage of resistance were Helicobacter pylori (metronidazole 50%-80%, clarithromycin 20%-40%, and levofloxacin 30%-35%), Clostridioides difficile (clindamycin 8.3%-100%, cephalosporines 51%), Campylobacter jejuni and Campylobacter coli (fluoroquinolones 85%), Escherichia coli (ampicillin 76.5%), Entamoeba histolytica (metronidazole 50%), and bacterial peritonitis (third-generation cephalosporines 40%, methicillin 85%). Antimicrobial resistance is reaching elevated percentages, making it necessary to evaluate the situation of each patient, to successfully treat gastrointestinal infections.


Insulin like growth factor binding protein 2 (IGFBP-2) for risk prediction in patients with severe aortic stenosis undergoing Transcatheter Aortic Valve Implantation (TAVI).
Severe aortic stenosis (AS) caused by degenerative calcification is the most frequent acquired valvular heart disease worldwide and mortality rates are considerably high. Transcatheter Aortic Valve Implantation (TAVI) is a well-established method for valve replacement in high risk patients with AS. However, there is a lack of reliable predictors for patients undergoing TAVI since commonly used scores were developed for surgical populations. 208 patients subjected to TAVI were included in this study. Plasma samples were obtained before TAVI and were evaluated for IGFBP-2 using commercially available ELISA kits. IGFBP-2 levels were analyzed for their ability for risk prediction after TAVI. IGFBP-2 levels measured before TAVI correlated significantly with left ventricular ejection fraction, EUROSCORE and other functional and prognostic parameters like the 6-minute walking test. When patients were retrospectively divided in two groups with a cut-off of serum IGFBP-2 levels of 275 ng/ml, IGFBP-2 was a strong predictor for 30-day and one-year mortality (3% vs. 11%, p = 0.05 and 18.2% vs. 46.2%; p < 0.001 respectively). Compared to an EUROSCORE above 20 or an STS score cut-off above 8, IGFBP-2 plasma levels above 275 ng/ml outperformed the established risk score for prediction of one-year mortality as assessed by NRI (0.65 95% CI 0.37-0.94; p < 0.001 and 0.54 95% CI 0.25-0.82; p < 0.001, respectively). Our results indicate that IGFBP-2 could serve as new outcome predictor for patients undergoing TAVI procedure. By providing additional information to the commonly used EUROSCORE, IGFPB-2 analysis could further assist Heart Team decision making.


microRNA-140-3p modulates invasiveness, motility, and extracellular matrix adhesion of breast cancer cells by targeting syndecan-4.
Syndecan-4, a predicted target of the microRNA miR-140-3p, plays an important role in multiple steps of tumor progression and is the second most abundant heparan sulfate proteoglycan produced by breast carcinoma cell lines. To investigate the potential functional relationship of miR-140-3p and syndecan-4, MDA-MB-231, SKBR3, and MCF-7 breast cancer (BC) cells were transiently transfected with pre-miR-140-3p, syndecan-4 small interfering RNAJ, or control reagents, respectively. Altered cell behavior was monitored by adhesion, migration, and invasion chamber assays. Moreover, the prognostic value of syndecan-4 was assessed by Kaplan-Maier Plotter analysis of gene expression data from tumor samples of 4929 patients. High expression of syndecan-4 was associated with better relapse-free survival in the whole collective of BC patients, but correlated with a worse survival in the subgroup of estrogen receptor negative and estrogen/progesterone-receptor negative patients. miR-140-3p expression was associated with improved survival irrespective of hormone receptor status. miR-140-3p overexpression induced posttranscriptional downregulation of syndecan-4, as demonstrated by quantitative real-time PCR (qPCR), flow cytometry, and luciferase assays, resulting in decreased BC cell migration and matrigel invasiveness. Furthermore, miR-140-3p overexpression and syndecan-4 silencing increased the adhesion of BC to fibronectin and laminin. qPCR analysis demonstrated that syndecan-4 silencing leads to altered gene expression of adhesion-related molecules, such as fibronectin and focal adhesion kinase, as well as in the gene expression of the proinvasive factors matrix metalloproteinase 2 and heparanase (also known as HPSE). We conclude that syndecan-4 is a novel target of miR-140-3p that regulates BC cell invasiveness and cell-matrix interactions in the tumor microenvironment.


Effect of Opuntia ficus-indica flowers maceration on quality and on heat stability of olive oil.
This study was focused on the evaluation of the quality and the oxidative stability of olive oil added with Opuntia ficus-indica flowers. Two different amounts of O. ficus-indica flowers were considered 5 and 15% (w/w). The olive oils were evaluated towards their quality, fatty acids profile, total phenol contents and thermal properties by differential scanning calorimetry. The oxidative stability was also monitored by employing the Rancimat and the oven test based on accelerating the oxidation process during storage. The addition of O. ficus-indica flowers induced an increase in free acidity values and a variation in fatty acids profile of olive oils but values remained under the limits required for an extra-virgin olive oil. The obtained olive oils were nutritionally enriched due to the increase in their phenols content. The oxidative stability was generally improved, mainly in olive oil enriched with 5% Opuntia ficus-indica flowers. These findings proved that this enriched olive oil could be considered as a product with a greater added value.


PROTOCOL: Interventions to improve the economic self-sufficiency of unemployed immigrants from non-Western countries.
This is a protocol for a Campbell Review. The objectives are as follows. This review systematically collects and synthesizes evidence from evaluations of causal effects of interventions designed to improve employment outcomes for non-Western immigrants. The review aims to answer the following questions: 1)Do interventions designed to improve the economic self-sufficiency for non-Western immigrants affect participants employment, use of cash assistance, income, or job retention?2)Do effects differ depending on programme content or populations served?


Doxycycline-induced gastrointestinal injury.
Doxycycline-induced gastric injury is a rarely recognized adverse effect of a common medication. Only 2 cases have previously described the distinctive capillary degeneration identified in gastric mucosa. We expanded on this by describing additional involved sites, endoscopic findings, and patient characteristics. Gastrointestinal biopsy materials for cases indexed with the word doxycycline were retrieved and the histology reviewed. The medical record was used to obtain clinical details. Three cases with biopsy materials were identified from the search, and doxycycline ingestion was confirmed. All patients' gastric biopsies had small vessel injury with fibrinoid material around the vessel, and 1 patient had similar changes in the duodenum. Endoscopic findings included fundic and pyloric erosions and ulcers. One patient had a normal endoscopy on follow-up after drug cessation. Confirmation and increased understanding of this drug-specific injury pattern are important for patient management, as cessation appears to result in symptom improvement and healing.


Polyamine transport systems in the LLC-PK1 renal epithelial established cell line.
LLC-PK1 cells were brought to a quiescent state by treatment with DL-2-difluoromethylornithine (DFMO), a specific inhibitor of L-ornithine decarboxylase (ODC). The inhibition of ODC, which is the key enzyme for polyamine synthesis, strongly reduced the cellular content of putrescine and spermidine. The cells resumed DNA-synthesis followed by mitosis when exogenous putrescine was added. DFMO treatment strongly stimulated the putrescine uptake capability. A kinetic analysis of the initial uptake rates revealed a saturable Na+-dependent and a saturable Na+-independent pathway on top of non-saturable diffusion. The stimulation by DFMO was exclusively due to an effect on the Vmax values of the saturable pathways. The Na+-dependent transporter had a higher affinity for putrescine (apparent Km = 4.7 +/- 0.7 microM) than the Na+-independent transporter (apparent Km = 29.8 +/- 3.5 microM). As a consequence, although the latter transporter had a higher Vmax, the Na+-dependent transport was more important at a physiological putrescine concentration. Putrescine uptake by both transporters was inhibited with similar relative affinities by spermidine, spermine as well as by the antileukemic agent, methylglyoxal bis(guanylhydrazone), but not by amino acids. The activity of the Na+-dependent transporter was very much dependent on SH-group reagents, whereas the Na+-independent transporter was not affected. Both transporters were inhibited by metabolic inhibitors and by ionophores but the Na+-dependent transporter was affected to a greater extent. For both transporters there was a down-regulation in response to exogenous putrescine. This suggests that the polyamine transporters in LLC-PK1 are adaptively regulated and may contribute to the regulation of the cellular polyamine level and cellular proliferation.


Biological variations, genetic polymorphisms and familial resemblance of TNF-alpha and IL-6 concentrations: STANISLAS cohort.
Cytokines are involved in the development of several inflammatory diseases and atherosclerosis. Their variations in healthy individuals are not well defined. The aims of this study were: firstly, to identify factors affecting biological variation of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha); secondly, to study their family resemblance; and thirdly, to evaluate the effect of two TNF-alpha (-308G/A and -238G/A) and two IL-6 polymorphisms (174G/C and -572G/C) on their corresponding circulating levels. A total of 171 healthy families selected from the STANISLAS cohort were studied. Age was negatively related to TNF-alpha concentrations in offspring only (both sons and daughters). Additionally, IL-6 and TNF-alpha levels were differently influenced by gender, white blood cells, tobacco consumption, and HDL-cholesterol level. A weak significant familial resemblance for TNF-alpha concentration was observed in siblings only. There was no significant familial resemblance for IL-6 levels. The TNF-alpha -308A allele was associated with decreased TNF-alpha concentrations in both offspring aged less than 18 and males without overweight (BMI<25 kg/m(2)). Fathers carrying the IL-6 -174CC genotype had higher IL-6 levels than those with the IL-6 -174G allele. Parents with the IL-6 -572GG genotype had higher IL-6 concentrations than the C allele carriers. In this sample of healthy families, plasma levels of IL-6 and TNF-alpha were differently affected by biological parameters including age, gender and smoking, and the impact of their respective polymorphisms was influenced by gender, age and BMI.


The choice of controls for providing validity and evidence in clinical research.
Selection of controls, a group of subjects who are identical to the treatment group in all aspects that affect the outcome except the intervention of interest, is a significant criterion for conducting a study in evidence-based medical research. Few studies emphasize the appropriate selection of control groups in the plastic surgery literature. The authors performed a literature search in Plastic and Reconstructive Surgery from January 1, 2010, through December 31, 2011, for studies in which controls were needed. The number of studies using a control group, control selection criteria, and the characteristics of the control populations were evaluated. Three hundred twenty-seven articles were obtained from our search using the keywords "case control studies" and "retrospective cohort studies." Among these studies, 121 articles were studies conducted in humans. All of these studies based on the study design required a control group, yet only 63 studies (52 percent) had a comparative control group. Of these studies, the authors found biases regarding the choice of controls, including selection bias, misclassification bias, and chronology bias. The authors' review shows that 48 percent of the studies published in Plastic and Reconstructive Surgery that were required to have a control group failed to incorporate a well-defined control group. Specific details pertaining to the methods used and the success obtained with those methods in recruiting controls need to be stated explicitly in the article to ensure uniformity and to support the validity of the research.


[Combination of propofol-sufentanil on somatosensory evoked potentials in surgery of the spine].
Most anaesthetics depress cortical somatosensory evoked potentials (CSEPs). However, the modification of CSEPs during total intravenous anaesthesia using propofol remaining still unknown, justified this trial. Open, prospective, clinical study. Nine consecutive patients requiring CSEPs monitoring for spine surgery, were studied. Anaesthesia was induced with propofol (2.5 mg.kg-1 then 10 mg.kg-1.h-1) and sufentanil (0.50 micrograms.kg-1 then 0.25 micrograms.kg-1.h-1). Maximum positive (P40) and negative (N50) waweform latencies, and the peak to peak amplitude of CSEPs (posterior tibial nerve stimulation, cortical recording), were recorded before induction, 30 min, 1 and 2 h after induction, and at the end of surgery. Data are means +/- SD. Duration of anaesthesia was 260 +/- 73 min. Propofol induced significant lengthening of CSEPS (P40: from 37 +/- 10 up to 41 +/- 11 ms; N50: from 45 +/- 11 up to 51 +/- 14 ms), and a significant decrease in amplitude (from 1.9 +/- 0.9 down to 0.8 +/- 0.4 microV), but these changes were stable from 30 min after the induction to the end of spine surgery. A motor response was obtained 29 +/- 14 min after the end of anaesthetic administration. Total intravenous anaesthesia with propofol and sufentanil induces a small but stable lengthening of CSEPs latency and a stable decrease of its amplitude, which enable an appropriate monitoring of CSEPs during spine surgery.


Research Note: Nondestructive detection of super grade chick embryos or hatchlings using near-infrared spectroscopy.
Some unresolved questions in poultry science were addressed: what determines the yield of chick embryos or hatchlings; what kind of influence does egg yolk content have on embryonic development; and how to detect eggs producing super grade chicks? Since the yolk acts as a vital energy and nutrient reservoir for embryos, we hypothesized that a higher yolk content of similar sizes eggs would play an important role in embryo or chick viability during incubation, as well as at hatch. As experimental sample, we used ROSS 308 (broiler line) and a nondestructive spectroscopic absorbance method. The influence of high yolk content to embryonic heartbeat and chick yield (i.e., chick weight/egg weight) were then investigated. Embryonic heartbeat signal was measured indirectly using a prototype near-infrared sensor during incubation period. A positive influence was found in both cases. Similar size eggs with higher yolk content were found to significantly (P-value < 0.05) promote higher chick yield at hatch. This methodology may have the potential to be used to precision poultry production system, ornithology, developmental, or evolutionary biology in the near future.


A review on l-ribose isomerases for the biocatalytic production of l-ribose and l-ribulose.
Presently, because of the extraordinary roles and potential applications, rare sugars turn into a focus point for countless researchers in the field of carbohydrates. l-ribose and l-ribulose are rare sugars and isomers of each other. This aldo and ketopentose are expensive but can be utilized as an antecedent for the manufacturing of various rare sugars and l-nucleoside analogue. The bioconversion approach turns into an excellent alternative method to l-ribulose and l-ribose production, as compared to the complex and lengthy chemical methods. The basic purpose of this research was to describe the importance of rare sugars in various fields and their easy production by using enzymatic methods. l-Ribose isomerase (L-RI) is an enzyme discovered by Tsuyoshi Shimonishi and Ken Izumori in 1996 from Acinetobacter sp. strain DL-28. L-RI structure was cupin-type-β-barrel shaped with a catalytic site between two β-sheets surrounded by metal ions. The crystal structures of the L-RI showed that it contains a homotetramer structure. Current review have concentrated on the sources, characteristics, applications, conclusions and future prospects including the potentials of l-ribose isomerase for the commercial production of l-ribose and l-ribulose. The MmL-RIse and CrL-RIse have the potential to produce the l-ribulose up to 32% and 31%, respectively. The CrL-RIse is highly stable as compared to other L-RIs. The results explained that the L-RIs have great potential in the production of rare sugars especially, l-ribose and l-ribulose, while the immobilization technique can enhance its functionality and properties. The present study precises the applications of L-RIs acquired from various sources for l-ribose and l-ribulose production.


Intracoronary bolus-only compared with intravenous bolus plus infusion of tirofiban application in patients with ST-elevation myocardial infarction undergoing primary percutaneous coronary intervention.
The aim of this pilot study was to compare intracoronary bolus-only with standard intravenous bolus plus maintenance infusion of tirofiban with respect to improvement in myocardial reperfusion after primary percutaneous coronary intervention (p-PCI). Changes in clinical practice may obviate the need for a maintenance infusion of small molecule glycoprotein IIb/IIIa inhibitors in current practice. Forty-nine patients undergoing p-PCI were randomized to either intracoronary bolus-only (n = 25) or intravenous bolus plus infusion (n = 24) of tirofiban. The primary end point was coronary hemodynamic indices of microvascular perfusion measured 4-5 days after p-PCI. The secondary end points were ST segment resolution at 90 min, the corrected TIMI frame count and myocardial blush grade. At 6 months, echocardiography and technetium-99m single-photon-emission computed tomography were performed. Microvascular perfusion did not differ significantly between the two treatment groups: index of microvascular resistance (27 ± 13 vs. 35 ± 15 U, P = 0.08) and coronary flow reserve (2.2 ± 0.7 vs. 1.9 ± 0.6, P = 0.25). The corrected TIMI frame counts assessed in the first (P = 0.13) and the second (P = 0.09) catheterization or the myocardial blush grades evaluated immediately (P = 0.23) and 4-5 days after MI (P = 1.00) were not significantly different between the two groups. At 6 months, there was no difference between the two groups in infarct size, left ventricular volumes, or ejection fraction. The standard intravenous bolus plus maintenance infusion of tirofiban in p-PCI is not superior to intracoronary bolus-only administration with respect to microvascular perfusion. Further, adequately powered randomized trials are warranted to evaluate the clinical outcomes associated with this strategy.


[Inversion of the sequence of surgery after neoadjuvant chemotherapy for synchronous liver metastases from colorectal cancer].
In some patients with colorectal cancer and synchronous liver metastases, chemotherapy and current combinations of chemotherapy allow the size of these metastases to be reduced so that they can be surgically resected. However, in many patients, the initial systematic treatment of the primary tumor is associated with growth of the metastases (which predict the patient's life expectancy). This metastatic growth contraindicates surgical treatment that might otherwise be curative. We report the case of a patient with advanced recto-colonic cancer, which responded well to chemotherapy given as neoadjuvant treatment prior to surgery, in which the hepatic metastases were resected before excision of the primary tumor.


Reaching women with instructions on misoprostol use in a Latin American country.
In Latin America, where restrictive laws limit women's access to safe abortion services, misoprostol is being used to induce abortions, often without correct information on dosage or regimens. This study in an unnamed Latin American country aimed to identify appropriate channels through which instructions on misoprostol use could be disseminated to women. In-depth interviews were carried out with physicians, pharmacy staff, women who had had safe abortions and women from the community, as well as focus group discussions with advocates of safe abortion. Participants considered physicians to be the most appropriate source of information and for ensuring confidentiality for both women and provider. Participants considered midwives, pharmacists and women's groups as appropriate referral points, but not providers of information. Legal restrictions and professional risk were the primary reasons why pharmacists rejected this role, though many are selling misoprostol. There was a general lack of information about misoprostol for medical abortion among both health professionals and women. Accurate information about misoprostol use for a wide range of gynaecologic and obstetric purposes, including induced abortion, could be incorporated into training and educational materials for physicians, midwives and other appropriate mid-level providers, as well as pharmacists. Without these alternative information channels, access to information would be severely restricted, especially in rural areas.


Identification of Potential Extractablesand Leachables in Cosmetic Plastic Packagingby Microchambers-Thermal Extraction and Pyrolysis-Gas Chromatography-Mass Spectrometry.
Most container-content interaction studies are carried out through migration tests on end products or simulants involving generally toxic solvents. This study was conducted with the aim of identifying potential leachables from materials used in cosmetic plastic packaging by using two approaches based on solvent-free extraction, i.e., solid-phase microextraction sampling and pyrolyzer/thermal desorption coupled with gas chromatography mass spectrometry. Volatile and semi-volatile intentionally and non-intentionally added substances were detected in seven packaging samples made of polypropylene, polyethylene, and styrene-acrylonitrile copolymer. Thirty-five compounds related to the polymers industry or packaging industry were identified, among them phthalates, alkanes, styrene, and cyanide derivates including degradation products, impurities, additives, plasticizers, and monomers. All except eight belong to the Cramer class I. These thermodesorption techniques are complementary to those used for migration tests.


Influence of tilt training on activation of the autonomic nervous system in patients with vasovagal syncope.
Tilt training is a new treatment for vasovagal syncope. Its therapeutic efficacy is thought to be the result of the desensitization of cardiopulmonary receptors, but it could be the influence of the tilt training on the activation of the autonomic nervous system as well. The study group consisted of 24 vasovagal patients (17 women and 7 men) aged 32.5 +/- 11.8 years. The diagnostic head-up tilt test was performed according to the Italian protocol with nitroglycerin if necessary. The monitoring head-up tilt test was performed according to the Westminster protocol without provocation, after 1 to 3 months of tilt training. Holter ECG recordings for HRV parameters (time and frequency domain) were obtained from selected 2-min intervals before, during and after the diagnostic and monitoring tilt test. The diagnostic test was positive in the passive phase in 6 and after provocation in 18 patients. During the training period no syncope occurred. Analysing the HRV parameters we demonstrated the following findings: I. mRR decreases immediately after assumption of a vertical position in both tests (diagnostic and monitoring) but in the diagnostic test its further decrease occurs earlier than in the monitoring test; 2. the absolute power of the HF component is greater in the early phase of tilt after tilt training than in the corresponding period in the diagnostic test. After a longer period of tilt training the activation of the sympathetic nervous system in response to the erect position is diminished.


Formulation matters! A spectroscopic and molecular dynamics investigation on the peptide CIGB552 as itself and in its therapeutical formulation.
Synthetic therapeutic peptides (STP) are intensively studied as new-generation drugs, characterized by high purity, biocompatibility, selectivity and stereochemical control. However, most of the studies are focussed on the bioactivity of STP without considering how the formulation actually used for therapy administration could alter the physico-chemical properties of the active principle. The aggregation properties of a 20-mer STP (Ac-His-Ala-Arg-Ile-Lys-D-Pro-Thr-Phe-Arg-Arg-D-Leu-Lys-Trp-Lys-Tyr-Lys-Gly-Lys-Phe-Trp-NH2 ), showing antitumor activity, were investigated by optical spectroscopy and atomic force microscopy imaging, as itself (CIGB552) and in its therapeutic formulation (CIGB552TF). It has found that the therapeutic formulation deeply affects the aggregation properties of the investigated peptide and the morphology of the aggregates formed on mica by deposition of CIGB552 and CIGB552TF millimolar solutions. Molecular dynamics simulations studied the first steps of CIGB552 aggregation under physiological ionic strength conditions (NaCl 150 mM), showing that peptide oligomers, from dimers to tetramers, are preferentially formed in this environment. Interestingly, cell viability assays performed on H-460 cell lines indicate a major antiproliferative activity of the peptide in its therapeutic formulation with respect to the peptide aqueous solution.


DPP3/CDK1 contributes to the progression of colorectal cancer through regulating cell proliferation, cell apoptosis, and cell migration.
At present, colorectal cancer (CRC) has become a serious threat to human health in the world. Dipeptidyl peptidase 3 (DPP3) is a zinc-dependent hydrolase that may be involved in several physiological processes. However, whether DPP3 affects the development and progression of CRC remains a mystery. This study is the first to demonstrate the role of DPP3 in CRC. Firstly, the results of immunohistochemistry analysis showed the upregulation of DPP3 in CRC tissues compared with normal tissues, which is statistically analyzed to be positively correlated with lymphatic metastasis, pathological stage, positive number of lymph nodes. Moreover, the high expression of DPP3 predicts poor prognosis in CRC patients. In addition, the results of cell dysfunction experiments clarified that the downregulation of DPP3 significantly inhibited cell proliferation, colony formation, cell migration, and promoted apoptosis in vitro. DPP3 depletion could induce cell apoptosis by upregulating the expression of BID, BIM, Caspase3, Caspase8, HSP60, p21, p27, p53, and SMAC. In addition, downregulation of DPP3 can reduce tumorigenicity of CRC cells in vivo. Furthermore, CDK1 is determined to be a downstream target of DPP3-mediated regulation of CRC by RNA-seq, qPCR, and WB. The interaction between DPP3 and CDK1 shows mutual regulation. Specifically, downregulation of DPP3 can accentuate the effects of CDK1 knockdown on the function of CRC cells. Overexpression of CDK1 alleviates the inhibitory effects of DPP3 knockdown in CRC cells. In summary, DPP3 has oncogene-like functions in the development and progression of CRC by targeting CDK1, which may be an effective molecular target for the prognosis and treatment of CRC.


Drugs involved in drug-facilitated crimes--part II: Drugs of abuse, prescription and over-the-counter medications. A review.
In recent years, there has been a notable increase in the number of reports of drug-facilitated crimes. Usually, individuals report that they were robbed or assaulted while incapacitated by drugs. Most often, these cases have involved drugs that have the ability to produce an effect that leaves the victim in a semiconscious or unconscious state. It is reasonable to assume that the purpose of drug-induced incapacitation is probably largely unchanged with time. This covers the full range of property offences (particularly theft) and crimes against the person (often sexual assault). What have changed are the drugs themselves: the number; type; their accessibility; effects and detection. This review describes the different aspects related to the involvement and use of drugs of abuse, as well as prescription and over-the counter medications in drug-facilitated crimes, which may help people working in this field to expand their knowledge in order to better understand the nature of these crimes or offences.


Controversies in Postoperative Irradiation of Oropharyngeal Cancer After Transoral Surgery.
Transoral surgery (TOS) is a novel technology whose adoption is expanding in the United States and other countries. TOS offers the possibility of a minimally invasive surgical approach to head and neck cancers. Its most frequent application has been in oropharyngeal cancers (OPC), of which most are associated with human papillomavirus (HPV). For HPV-associated OPC, where high response and survival rates are expected, deintensification of standard therapy is a major area of clinical research. In HPV-OPC, traditional pathologic risk factors indicating a need for adjuvant radiation or chemoradiation may not apply as strongly.


Practical implications for the administration of 13-cis retinoic acid in pediatric oncology.
Children with high-risk neuroblastoma are treated with polychemotherapy, surgery, radiotherapy and even autologous stem-cell transplantation. On top of this complex treatment, most children also receive 13-cis retinoic acid as differentiation agent. As no suitable pharmaceutical formulation is available so far, there are often problems with the administration of the product in children. The present report describes some practical recommendations for the administration of isotretinoin in children treated for high-risk neuroblastoma.


Novel circular RNA circSLIT2 facilitates the aerobic glycolysis of pancreatic ductal adenocarcinoma via miR-510-5p/c-Myc/LDHA axis.
Increasing evidence has indicated the great diagnostic and therapeutic potentials of circular RNAs (circRNAs) in human cancers. Although the biological roles of circRNAs in pancreatic ductal adenocarcinoma (PDAC) have been partially annotated, the potential regulatory mechanism of circRNAs in PDAC tumorigenesis remains poorly understood. Here, our study found that the novel circRNA circSLIT2 was significantly upregulated in PDAC tissues and cells. Clinically, ectopic high-expression of circSLIT2 was correlated with unfavorable prognosis of PDAC patients. Functional experiments demonstrated that circSLIT2 promoted the aerobic glycolysis and proliferation of PDAC cells in vitro, and circSLIT2 knockdown inhibited tumor growth in vivo. Mechanistically, circSLIT2 acted as miRNA sponge to target miR-510-5p/c-Myc axis. Furthermore, c-Myc bound with the promoter region of lactate dehydrogenase A (LDHA) to activate the transcription. Collectively, present findings reveal that circSLIT2/miR-510-5p/c-Myc/LDHA axis participates in the aerobic glycolysis and carcinogenesis of PDAC, and may act as a promising therapeutic target.


Neutrophil-to-Lymphocyte Ratio Is Superior to Other Leukocyte-Based Ratios as a Prognostic Predictor in Non-Small Cell Lung Cancer Patients with Radiosurgically Treated Brain Metastases Under Immunotherapy or Targeted Therapy.
To investigate predictive value of preradiosurgery leukocyte-based prognostic ratios in a selected cohort of non-small cell lung cancer (NSCLC) patients with radiosurgery-treated brain metastases (BM) and concomitant immunotherapy (IT) or targeted therapy (TT). We performed a retrospective analysis of 166 patients with NSCLC BM treated with Gamma Knife radiosurgery. Neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio, and lymphocyte-to-monocyte ratio were assessed within 14 days before radiosurgery. In radiosurgically treated patients with NSCLC BM with concomitant IT or TT, estimated median survival after first Gamma Knife radiosurgery treatment was significantly longer in patients with NLR cutoff value <5 (P = 0.038). Consequently, the Cox regression model for NLR cutoff value groups revealed a significant hazard ratio of 1.519 (95% confidence interval 1.020-2.265, P = 0.040). In addition, each increase in NLR of 1 equaled an increase of 5.4% in risk of death (hazard ratio 1.054, 95% confidence interval 1.024-1.085, P < 0.001). After adjusting for sex, age, Karnofsky performance scale, and presence of extracranial metastases, NLR remained a significant and independent predictor for survival (hazard ratio 1.047, 95% confidence interval 1.017-1.078, P = 0.002). In contrast, platelet-to-lymphocyte ratio and lymphocyte-to-monocyte ratio did not exhibit the same predictive value among patients with radiosurgery-treated BM with concomitant IT or TT. In patients with NSCLC BM treated with radiosurgery with concomitant IT or TT, preradiosurgery NLR represents a simple prognostic predictor for survival and is superior to other leukocyte-based ratios. NLR may be relevant for clinical decision making, therapeutic evaluation, patient counseling, and appropriate stratification of future clinical trials among patients with radiosurgery-treated BM.


Macrophage activation by a vanadyl-aspirin complex is dependent on L-type calcium channel and the generation of nitric oxide.
Bone homeostasis is the result of a tight balance between bone resorption and bone formation where macrophage activation is believed to contribute to bone resorption. We have previously shown that a vanadyl(IV)-aspirin complex (VOAspi) regulates cell proliferation and differentiation of osteoblasts in culture. In this study, we assessed VOAspi and VO effects and their possible mechanism of action on a mouse macrophage cell line RAW 264.7. Both vanadium compounds inhibited cell proliferation in a dose-dependent manner. Nifedipine completely reversed the VOAspi-induced macrophage cytotoxicity, while it could not block the effect of VO. VOAspi also stimulated nitric oxide (NO) production, the oxidation of dihydrorhodamine 123 (DHR-123) and enhanced the expression of both constitutive and inducible isoforms of nitric oxide syntases (NOS). All these effects were abolished by nifedipine. Altogether our finding give evidence that VOAspi-induced macrophage cytotoxicity is dependent on L-type calcium channel and the generation of NO though the induction of eNOS and iNOS. Contrary, the parent compound VO exerted a cytotoxic effect by mechanisms independent of a calcium entry and the NO/NOS activation.


Association of pain management and positive expectations with psychological distress and spiritual well‑being among terminally ill cancer patients admitted to a palliative care unit.
Although palliation of psycho-spiritual distress is of great importance in terminally ill cancer patients, there is a little information about screening patients who benefit from palliative care and identifying the cancer care targets. This study explored the relationship of pain management and positive expectations with depression, anxiety and spiritual well-being (SWB) in terminal cancer patients admitted to a palliative care unit. Eighty-four terminal cancer inpatients were recruited from the Hospice Ward, Shengjing Hospital of China Medical University. Optimism and general self-efficacy (GSE) were evaluated at admission. Patients completed self-report questionnaires on SWB, depression, anxiety and pain both on admission and one week later. The repeated designed analysis of variance was used to explore the correlates of depression, anxiety and SWB (meaning, peace, faith). In our sample, only cancer pain diminished significantly one week later. For depression (p = 0.041) and faith (p = 0.013), there was a significant pain group (relieved vs. not relieved) × time interaction effect, such that those with satisfied pain control experienced the improved psycho-spiritual outcomes at 1 week. The relationship between positive expectations, peace and faith was also statistically significant, indicating that the improvement of peace or faith was significant in the low group of optimism and GSE. Our findings indicated that pain management lied at the center of depression and SWB, meaning that effective pain management may reduce depression, and improve SWB among terminal cancer patients. Moreover, positive expectations, especially for optimism, may be the new target for SWB-related intervention research. Palliative care nurse should require the identification of terminal cancer patients who may more benefit from short-term palliative care, and target them with effective cancer care.


The evolution of a case of overlap syndrome with systemic sclerosis, rheumatoid arthritis and systemic lupus erythematosus.
An unusual case of overlap syndrome which evolved over a 12-year period is described. The patient initially presented with limited cutaneous systemic sclerosis. She then developed seropositive erosive rheumatoid arthritis and subsequently vasculitis with positive lupus serology. There was no evidence that she had mixed or undifferentiated connective tissue disease, and antibody to ribonuclearprotein was negative. This unusual combination of connective tissue disorders in one patient is reported and the literature is reviewed.


Effect of antioxidant, malondialdehyde, macro-mineral, and trace element serum concentrations in Bangladeshi patients with schizophrenia: A case-control study.
Schizophrenia (SCZ) is an incurable neuropsychiatric disorder generally described by impaired social behavior and altered recognition of reality. For the first time, this study explored serum levels of antioxidants (vitamin A, E, and C), malondialdehyde (MDA), macro-minerals (calcium, potassium, and sodium), and trace elements (zinc, iron, and selenium) in Bangladeshi patients with SCZ and thereby, discovering any pathophysiological correlation. This case-controlled study evaluated 63 patients with SCZ as cases and 63 healthy individuals as controls. Vitamin A and E levels were defined by RP-HPLC. MDA and vitamin C levels were measured by using UV spectrophotometry, and macro and trace elements by atomic absorption spectroscopy. This study found significantly (P ≤ 0.05) elevated MDA levels and decreased levels of antioxidants-vitamin A, C, and E and significantly (P ≤ 0.05) diminished levels of macro and trace elements in cases in contrast to the controls. Serum levels of zinc (Zn), selenium (Se), iron (Fe), potassium (K), calcium (Ca), and sodium (Na) were determined to be 0.33 ± 0.008, 0.0252 ± 0.00060, 0.24 ± 0.01, 64.18 ± 2.72, 36.88 ± 2.56, and 2657.5 ± 53.32 mg/L, respectively, in cases, whereas 0.79 ± 0.03, 0.0650 ± 0.00355,0.78 ± 0.03, 168.01 ± 2.85, 86.43 ± 2.55, and 3200.8 ± 29.96 mg/L, respectively, were determined in controls. Pearson's correlation analysis revealed a negative correlation between Zn and Na, Zn and K, Zn and Ca, Zn and Fe, Zn and Se, Fe and Na, and Fe and Se in patients. The findings connect that the pathogenesis of SCZ may have a correlation with altered levels of antioxidants, MDA, macro-minerals, and trace elements.


Gastrointestinal adverse reactions reduce the success rate of Helicobacter pylori eradication therapy: A multicenter prospective cohort study.
The screening and treatment of Helicobacter pylori infection for all junior high students in Saga Prefecture, Japan, were started in 2016. The present study aims to evaluate the influence of adverse reactions on the success of the eradication therapy. From 2017 to 2019, 25,006 third-grade junior high school students were tested for urinary anti-H. pylori antibodies. Positive cases were confirmed by H. pylori stool antigen tests. Of the 531 students who were found to be H. pylori-positive, 390 (358 in first-line and 32 in second-line therapy) underwent eradication therapy, and 274 (242 in first-line and 32 in second-line) students actually completed a self-reported form to rate stool consistency (based on the Bristol Stool Scale), the maximum number of bowel movements, and abdominal symptoms during the 7 days of treatment. Among the 274 students, the total of primary and secondary eradication success rates was 87% (95% confidential interval: 82.9-90.1) in intention-to-treat analysis. On days 4, 5, and 6, stool consistency was looser in the primary eradication failure group than in the success group (p < .05). Looser stool consistencies were observed in male students with abdominal pain compared to those who did not experience pain (p < .05). Abdominal pain and diarrhea were detected in 28.5% and 42.7% of the subjects, respectively. The overall incidence of other adverse events was low (n = 8/274, 2.9%), and only two students discontinued treatment because of adverse events. Softening of the stool was related to the eradication failure in the junior high school students, especially in males with abdominal pain. Adverse effects did not induce discontinuation of the eradication treatment.


Water and sectoral policies in agriculture-forest frontiers: An expanded interdisciplinary research approach.
Major land use changes such as deforestation and restoration influence water resources in agriculture-forest landscapes. Changes are observed in water flows, groundwater infiltration, water quality and rainfall. Interdisciplinary water-forest research has unravelled biophysical parts of the interplay that influences forest and water resources. In this Perspective paper, we propose an expanded interdisciplinary research approach to study water and policies in agriculture-forest frontiers. The approach differs in four important aspects from previous ones: (i) a conceptual 'frontier' understanding; an analytical focus on (ii) agriculture and (iii) policy-water linkages; (iv) empirical attention to northern and southern countries. The approach is put into practice with the "Pendulum" framework, with interventions and the agriculture-forest frontier oscillating over time between exploitation and restoration. Through the approach, a better understanding will be provided on the dynamic interplay of water and policies in oscillating agriculture-forest frontiers, with changing outcomes for people and environment.


Combining clinical exams can better predict lumbar spine radiographic instability.
Several clinical tests have been proposed to diagnose lumbar instability, but their accuracy is still in question. The primary purpose of this study was to evaluate the diagnostic accuracy of the clinical lumbar instability tests. The secondary goal was to design a model to detect lumbar instability. A prospective diagnostic cross-sectional study. A sample of 202 patients with chronic low back pain were participated in the study. Five lumbar instability tests including Aberrant movement, Passive lumbar extension, Prone segmental instability, H and I and pheasant tests were compared to flexion/extension radiography as the gold standard for diagnosing lumbar instability using two by two tables. Multiple Logistic Regression analysis was applied to develop a model using demographic information as well as the patients' pain intensity, disability level, lumbar lordosis and the clinical tests. Among the five examined tests, Prone segmental instability, H and I and pheasant tests showed very small likelihood ratios and diagnostic odd's ratio. The largest values were for H and I test with the positive likelihood ratio of 1.28 (95% CI: 0.72 to 2.29) and diagnostic odd's ratio of 1.37 (95% CI: 0.66 to 2.83); the diagnostic accuracy measures were smaller for the other studied clinical tests. The model was developed using weight (t = 1.15, p = 0.03) and lumbar lordosis (t = 3.04, p = 0.00) (which showed a significant relationship with lumbar instability) and prone segmental instability test. The final model has the positive likelihood ratio of 2.07 (95% CI: 1.41 to 3.05) and diagnostic odd's ratio of 3.77 (95% CI: 2.03 to 7.01). Each individual test had very small to no power in discriminating patients with lumbar instability. The developed model just slightly improved the accuracy of radiological instability detection.


Impact of acute mental stress on segmental arterial stiffness.
It has been reported that acute brief episodes of mental stress (MS) result in a prolonged increase in carotid-femoral pulse wave velocity (cfPWV), an index of aortic stiffness. However, whether acute MS also impacts arterial stiffness in other segments is unclear. The present study aimed to examine the impact of acute MS on segmental arterial stiffness. In the main experiment, 17 young male subjects (mean age, 20.1 ± 0.7 years) performed a 5-min MS and control (CON) task in a random order. Pulse wave velocity (PWV) from the heart to the brachium (hbPWV) and the ankle (haPWV), PWV between the brachial artery and the ankle (baPWV), and the cardio-ankle vascular index (CAVI) were simultaneously measured at baseline and 5, 15, and 30 min after the task. Compared to baseline values, hbPWV, baPWV, haPWV, and CAVI significantly increased until 30 min after the MS task, whereas these variables did not significantly change following the CON task. At 5 and 30 min after the MS task, percentage changes from baseline were significantly higher in hbPWV (+ 5.2 ± 4.4 and 6.6 ± 4.9%) than in baPWV (+ 2.2 ± 2.1 and 2.2 ± 2.0%) or haPWV (+ 3.6 ± 2.6 and 4.3 ± 2.9%) and were also significantly lower in baPWV than in haPWV. These findings suggest that acute MS elicits an increase in arterial stiffness in various segments and this arterial stiffening is not uniform among the segments.


Construction of C-C bonds via photoreductive coupling of ketones and aldehydes in the metal-organic-framework MFM-300(Cr).
Construction of C-C bonds via reductive coupling of aldehydes and ketones is hindered by the highly negative reduction potential of these carbonyl substrates, particularly ketones, and this renders the formation of ketyl radicals extremely endergonic. Here, we report the efficient activation of carbonyl compounds by the formation of specific host-guest interactions in a hydroxyl-decorated porous photocatalyst. MFM-300(Cr) exhibits a band gap of 1.75 eV and shows excellent catalytic activity and stability towards the photoreductive coupling of 30 different aldehydes and ketones to the corresponding 1,2-diols at room temperature. Synchrotron X-ray diffraction and electron paramagnetic resonance spectroscopy confirm the generation of ketyl radicals via confinement within MFM-300(Cr). This protocol removes simultaneously the need for a precious metal-based photocatalyst or for amine-based sacrificial agents for the photochemical synthesis.


Clinical Presentation and Surgical Outcomes Based on Age and Tumor Topography in 59 Patients With Spinal Dermoid Cysts.
Differences in presentation and outcomes in children and adults with spinal dermoid tumors have not been documented. Surgical outcomes for dermoid tumors in different spinal cord planes have also not been studied. Retrospective review of 59 consecutive patients who underwent surgery for intraspinal dermoid tumors at a single institution. Tumors were categorized as purely extramedullary (n = 15), purely intramedullary (n = 28) or intramedullary-extramedullary (n = 16) based on intraoperative and imaging findings. The cohort was composed of 45 children and 14 adults. The presence of a dermal sinus and prior history of meningitis was significantly associated with childhood presentation (P = 0.001 and P = 0.013, respectively). Most adults had purely intramedullary tumors (n = 12, 85.7%) as compared to children (35.6%, P = 0.001). Gross total resection (GTR) was achieved in 33.3% and 41.7% of children and adults, respectively (P = 1.000). Factors associated with GTR on logistic regression analysis included short segment (≤3 levels) dermoid cysts (P = 0.037) and absence of an intramedullary component (P = 0.027). Forty-seven patients were followed up over a mean period of 72.9 ± 8.3 months. Preoperative motor deficits were more likely to improve in children than in adults (P = 0.005). Recurrences were seen in 14.9% of cases (all with intramedullary tumors) at a median interval of 58 months (range: 40-166 months) after surgery, with no differences in recurrence-free survival between children and adults (P = 0.936). Presentation of dermoid cysts is distinctively different in children and adults. GTR is a challenge for intramedullary and large tumors. Recurrences may manifest several years after surgery, mandating life-long surveillance.


The sodium-glucose co-transporter-2 inhibitor ertugliflozin modifies the signature of cardiac substrate metabolism and reduces cardiac mTOR signalling, endoplasmic reticulum stress and apoptosis.
To investigate cardiac signalling pathways connecting substrate utilization with left ventricular remodelling in a murine pressure overload model. Cardiac hypertrophy was induced by transverse aortic constriction surgery in 20-week-old C57BL/6J mice treated with or without the sodium-glucose co-transporter 2 (SGLT2) inhibitor ertugliflozin (225 mg kg-1 chow diet) for 10 weeks. Ertugliflozin improved left ventricular function and reduced myocardial fibrosis. This occurred simultaneously with a fasting-like response characterized by improved glucose tolerance and increased ketone body concentrations. While cardiac insulin signalling was reduced in response to SGLT2 inhibition, AMP-activated protein kinase (AMPK) signalling was increased with induction of the fatty acid transporter cluster of differentiation 36 and phosphorylation of acetyl-CoA carboxylase (ACC). Further, enzymes responsible for ketone body catabolism (β-hydroxybutyrate dehydrogenase, succinyl-CoA:3-oxoacid-CoA transferase and acetyl-CoA acetyltransferase 1) were induced by SGLT2 inhibition. Ertugliflozin led to more cardiac abundance of fatty acids, tricarboxylic acid cycle metabolites and ATP. Downstream mechanistic target of rapamycin (mTOR) pathway, relevant for protein synthesis, cardiac hypertrophy and adverse cardiac remodelling, was reduced by SGLT2 inhibition, with alleviation of endoplasmic reticulum (ER) stress and unfolded protein response (UPR) providing a potential mechanism for abundant reduced left ventricular apoptosis and fibrosis. SGLT2 inhibition reduced left ventricular fibrosis in a murine model of cardiac hypertrophy. Mechanistically, this was associated with reduced cardiac insulin and increased AMPK signalling as a potential mechanism for less cardiac mTOR activation with alleviation of downstream ER stress, UPR and apoptosis.


Prenatal ultrasound diagnosis of a rare occurrence of lethal multiple pterygium syndrome in two siblings.
We report two cases of prenatal ultrasound diagnosis of the lethal form of multiple pterygium syndrome in successive pregnancies in the same woman. This extremely rare entity is characterized by multiple pterygia, flexion contractures, facial anomalies and hypoplasia of heart and lungs. The appearance of the congenital cataract is unique and this finding was present in both abortuses. Prenatal detection of this syndrome is feasible, and an ultrasound survey should be offered during genetic counselling to previously affected couples.


Single cell spectroscopy of red blood cells in intravenous crystalloid fluids.
Crystalloid fluids, a subset of intravenous (IV) fluid solutions are commonly used in clinical settings. The influence of these fluids on the functions of blood components are least explored. Raman spectroscopy combined with optical trapping has been widely used to evaluate the impact of external stress agents on red blood cells. The present study investigates the impact of commonly used crystalloid fluids on red blood cells in comparison with that of blood plasma using Raman Tweezers spectroscopy. The red blood cells suspended in crystalloid fluids undergo deoxygenation readily than that in blood plasma. In addition, cells in blood plasma were able to withstand laser induced deoxygenation comparatively better than that in crystalloid fluids at higher laser powers. Principle component analysis of the Raman spectral data has clearly demonstrated the discrimination of cells in plasma with that of crystalloid fluids demonstrating the effect of external induced stress on RBCs.


[Features of the clinical picture, diagnosis, and treatment of cranio-cerebral injuries with occipital bone fractures].
Contusion of the brain at the site of the injury and by the contrecoup mechanism as well as vascular dysfunction in the vertebrobasilar basin may occur in craniocerebral injury with fracture of the occipital bone. In view of this, it is necessary to analyse the neurological manifestations of the brain injury carefully, carry out radiography of the occipital bone, and rule out the presence of hematoma at the site of the injury and one of a contrecoup mechanism. Procaine hydrochloride block of points of the vertebral arteries in indicated because of the dysfunction in the vertebrobasilar basin.


Enhanced aqueous Cr(VI) removal using chitosan-modified magnetic biochars derived from bamboo residues.
The purpose of this study was to fabricate a chitosan modified magnetic bamboo biochar (CMBB) for Cr(VI) detoxification from aqueous solution. Results showed that chitosan modification provided more active adsorption sites on the surface of magnetic bamboo biochar (MBB), and hence enhanced Cr(VI) removal from aqueous solution. The maximum adsorption capacities of MBB and CMBB for Cr(VI) at 25 °C were 75.8 and 127 mg g-1, respectively. Increasing solution pH inbibited Cr(VI) adsorption by adsorbents. However, CMBB maintained a high Cr(VI) removal efficiencies over a broader pH range (2-10), and could attain 36% of the maximum adsorption (40 mg g-1) even at a high pH of 10.0. Rising temperature enhanced the Cr(VI) removal by two adsorbents. The escalating ionic strength and coexist substances, including Na+, Ca2+, Fe3+, Cl-, SO42-, PO43- and humate, inhibited the adsorption efficiency of Cr(VI) on adsorbents. After the fifth adsorption-desorption cycle, the adsorption efficiencies of CMBB and MBB for Cr(VI) remained above 90% and less than 50%, respectively. All these results indicated that CMBB could be a practical adsorbent that can be utilized for the detoxification of Cr(VI) ions from wastewater.


Incidence and risk factors in the appearance of heterotopic ossification in spinal cord injury.
Heterotopic ossification (HO) is a frequent complication in patients with a spinal cord injury (SCI), although the aetiology is unknown. A study was undertaken of 654 SCI patients with traumatic aetiology, admitted for the first time to the Hospital Nacional de Paraplejicos, Toledo, during 1988 and 1989. Of the total number of patients, 85 (13%) were diagnosed HO and 569 without HO. The diagnosis was mainly achieved by x-ray studies and clinical signs. From the 569 patients with traumatic aetiology without HO, 44 were selected at random, as were 44 of the 85 patients with HO. The mean time lapse between the occurrence of the accident and admission for patients with HO was 40.79 days (typical deviation (TD) = 45.2), and for patients without HO was 32.84 (TD = 38) days, resulting in a value of F = 0.796 through analysis of variance, which is not a statistically significant variation between the 2 groups. In both groups we have taken account of the following variables: age at time of lesion, lesion level, type of lesion (complete or incomplete), spasticity, urinary tract complications, deep vein thrombosis, important associated injuries occurring at the moment of lesion, time elapsed before admission and the existence of pressure sores. In those SCI patients with HO the number of ossifications and their localisations were also verified. By use of the chi square test (X2) over all 9 variables which were studied, we found that 3 variables (complete spinal lesion, presence of pressure sores and spasticity) were significantly related to HO formation.(ABSTRACT TRUNCATED AT 250 WORDS)


How to approach the acute respiratory distress syndrome: Prevention, plan, and prudence.
The acute respiratory distress syndrome (ARDS) is typically manifested by refractory hypoxemia with high mortality. A correct diagnosis is the first step to achieve better outcomes. An early intervention to manage modifiable risk factors of ARDS development and the avoidance of aggravating factors that increase disease severity and progression should be carefully addressed. A management plan is necessary at an early stage of ARDS to determine the level of intensive care. It should be carefully decided which therapeutic measures should be performed depending on the patient׳s underlying clinical condition. The clinician׳s considerate prudence is required in decisions of when to apply intensive measures for an ARDS treatment. Mechanical ventilator support should be carefully used depending on the patient׳s severity and pathological phase. Decreasing inappropriate alveolar strain through a low tidal volume under optimal positive end-expiratory pressure is key for ventilator support in ARDS. The extracorporeal membrane oxygenation applied in the experienced centers seems to improve the survival of patients with severe ARDS. A constellation of physical and psychological problems can develop or persist for up to 5 years in patients with ARDS. Therefore, an early mobilization with rehabilitation, even during an intensive care unit stay, should be seriously considered whenever feasible. Lastly, prevention of aspiration, stress ulcers, deep vein thrombosis, catheter-related infection, overhydration, and heavy sedation is essential to achieve better outcomes in ARDS.


Studies on the mechanism of membrane fusion. Role of head-group composition in calcium- and magnesium-induced fusion of mixed phospholipid vesicles.
We have investigated the contribution of various phospholipids to membrane fusion induced by divalent cations. Fusion was followed by means of a new fluorescence assay monitoring the mixing of internal aqueous contents of large (0.1 micrometer diameter) unilamellar liposomes. The rate and extent of fusion induced by Ca2+ in mixed phosphatidylserine/phosphatidylcholine vesicles were lower compared to those in pure phosphatidylserine vesicles. The presence of 50% phosphatidylcholine completely inhibited fusion, although the vesicles aggregated upon Ca2+ addition. When phosphatidylserine was mixed with phosphatidylethanolamine, however, rapid fusion could be induced by Ca2+ even in mixtures that contained only 25% phosphatidylserine. Phosphatidylethanolamine also facilitated fusion by Mg2+ which could not fuse pure phosphatidylserine vesicles. In phosphatidylserine/phosphatidylethanolamine/phosphatidylcholine mixtures, in which the phosphatidylcholine content was kept at 25%, phosphatidylethanolamine could not substitute for phosphatidylserine, and the fusogenic capacity of Mg2+ was abolished by the presence of merely 10% phosphatidylcholine. The initial rate of release of vesicle contents was slower than the rate of fusion in all the mixtures used. The presence of phosphate effected a considerable decrease in the threshold concentration of Ca2+ and also enhanced the rate and extent of fusion. Mg2+ had a synergistic effect on Ca2+-induced fusion of phosphatidylserine/phosphatidylethanolamine vesicles. We suggest that the role of phospholipids in membrane fusion is related to their ability to form dehydrated intermembrane complexes with divalent cations.


Advances in MRI Methodology.
Magnetic resonance imaging (MRI) is a non-invasive technique which can provide comprehensive, multi-parametric information on brain anatomy, function and metabolism. This chapter will give a brief introduction to the basic principles of MR and cover the evolution of image acquisition. Given the versatility of MR, a vast range of MR applications will be discussed, including structural, diffusion tensor, functional, perfusion and neuromelanin-sensitive imaging, as well as quantitative susceptibility mapping. This chapter will also cover methodological developments in MRI, including image analysis approaches that can be tailored according to experimental design or aims. The ultimate goal of this chapter is to equip readers with a fundamental understanding of the proficiencies and limitations of MR techniques and their corresponding analysis approaches.


Aversive view memories and risk perception in navigating ants.
Many ants establish foraging routes through learning views of the visual panorama. Route models have focused primarily on attractive view use, which experienced foragers orient towards to return to known sites. However, aversive views have recently been uncovered as a key component of route learning. Here, Cataglyphis velox rapidly learned aversive views, when associated with a negative outcome, a period of captivity in vegetation, triggering increases in hesitation behavior. These memories were based on the accumulation of experiences over multiple trips with each new experience regulating forager hesitancy. Foragers were also sensitive to captivity time differences, suggesting they possess some mechanism to quantify duration. Finally, we analyzed foragers' perception of risky (i.e. variable) versus stable aversive outcomes by associating two sites along the route with distinct captivity schedules, a fixed or variable duration, with the same mean across training. Foragers exhibited fewer hesitations in response to risky outcomes compared to fixed ones, indicating they perceived risky outcomes as less severe. Results align with a logarithmic relationship between captivity duration and hesitations, suggesting that aversive stimulus perception is a logarithm of its actual value. We discuss how aversive view learning could be executed within the mushroom bodies circuitry following a prediction error rule.


Further evidence for the anti-inflammatory activity of oleocanthal: inhibition of MIP-1α and IL-6 in J774 macrophages and in ATDC5 chondrocytes.
Given the relevance of degenerative joint diseases in our society, the development of a novel pharmacologic intervention is a critically important public health goal. Recently, oleocanthal, a polyphenolic natural compound from extra virgin olive oil, has emerged as a potential therapeutic weapon for the treatment of inflammatory degenerative diseases. The goal of this study was to further evaluate the anti-inflammatory activity of oleocanthal in murine macrophages J774 and murine chondrocytes ATDC5 with a particular focus on the inhibition of gene expression of pro-inflammatory factors such as MIP-1α and IL-6. ATDC5 murine chondrogenic cells and murine macrophages J774 were used. J774 macrophages were tested with different doses of oleocanthal and cell viability was evaluated using the MTT assay. Western blot analysis was carried on in J774 cells using anti NOS2 antibody. Nitrite accumulation was determined in culture supernatant using the Griess reaction. MIP-1α and IL-6 mRNA levels were determined using SYBR Green-based quantitative RT-PCR. MIP-1α and IL-6 protein levels were evaluated using specific ELISA assay. Several cytokines, involved in the inflammatory response, were also tested by BioPlex assay. First, oleocanthal inhibits LPS-induced NO production in J774 macrophages, without affecting cell viability. Moreover, it inhibits MIP-1α and IL-6 mRNA expression, as well as protein synthesis, in both ATDC5 chondrocytes and J774 macrophages. Oleocanthal also inhibits IL-1β, TNF-α and GM-CSF protein synthesis from LPS-stimulated macrophages. Our data confirm a clear potent role of oleocanthal as anti-inflammatory therapeutic agent for future treatment of arthritis or other inflammatory diseases.


Effect of socioeconomic position on patient outcome after hysterectomy.
To investigate the association between socioeconomic position (assessed by education, employment and income) and complications following hysterectomy and assess the role of lifestyle, co-morbidity and clinical conditions on the relationship. Register-based cohort study. The study included nearly all Danish women (n = 22 150) registered with a benign elective hysterectomy in the Danish Hysterectomy Database in 2004-2008. Data were analyzed using logistic regression models estimating the odds ratio with 95% confidence intervals. Complications following hysterectomy. Seventeen percent of the women experienced complications in relation to the hysterectomy. Women with less than high school education and unemployed women had higher odds of infection, complications and readmission than women with more than high school education and employed women. Furthermore, unemployed women had higher odds of hospitalization >4 days than women in employment. Lifestyle factors (smoking and body mass index) and co-morbidity status seemed to explain most of the social differences. However, an association between women with less than high school education and all complications remained unexplained. Furthermore, differences in lifestyle and co-morbidity status only partially explained the higher odds of infection, complications and hospitalization >4 days for unemployed than employed women. Women with a low socioeconomic position have significantly higher odds of complications following hysterectomy compared with women with a high socioeconomic position. Unhealthy lifestyle and presence of co-morbidity in women with low socioeconomic position partially explains the differences in complications.


Delay in diagnosis of primary osteosarcoma of bone in children: Have we improved in the last 15 years and what is the impact of delay on diagnosis?
Examine: (1) If length of symptoms (LS) of children with osteosarcoma has improved in the last 15 years (2) Is delay in diagnosis related to the presence of metastases at presentation? (3) The impact of delay in diagnosis on prognosis. 250 consecutive patients with a diagnosis of osteosarcoma of bone treated at a national bone tumor treatment center between 2004 and 2018 were studied retrospectively. Three groups comprising those diagnosed over a five-year period (Group 1: 2004-2008, Group 2: 2009-2013; Group 3: 2014-2018) were studied. There were 126 males and 124 females with a mean age 12.2 years. The median LS for all patients was eight weeks. The median LS for Group 3 was significantly shorter than that for other groups. Development of metastasis during follow-up period was significantly less in Group 3 compared to the other groups. Overall survival gradually improved over the whole study period. There was no difference in the proportion presenting with metastases at diagnosis between the three groups. The survival rates in patients with LS shorter than 4 weeks was better than those with LS longer than 4 weeks, irrespective of the study time period. There has been an improvement in the LS in patients diagnosed with osteosarcoma over the last 15 years. The development of metastasis during follow-up has reduced and the overall survival in the last 15 years has improved. LS longer than 4 weeks is associated with a poorer prognosis.


Epigenetic programming-The important first 1000 days.
The perinatal period is a time of fast physiological change, including epigenetic programming. Adverse events may lead to epigenetic changes, with implications for health and disease. Our review covers the basics of clinical epigenetics and explores the latest research, including the role of epigenetic processes in complex disease phenotypes, such as neurodevelopmental, neurodegenerative and immunological disorders. Some studies suggest that epigenetic alterations are linked to early life environmental stressors, including mode of delivery, famine, psychosocial stress, severe institutional deprivation and childhood abuse. CONCLUSION: Epigenetic modifications due to perinatal environmental exposures can lead to lifelong, but potentially reversible, phenotypic alterations and disease.


Synthesis of poly(A)-associated RNA in the prereplicative phase of isoproterenol-stimulated DNA synthesis.
Rapidly labelled RNA containing poly(A) sequences has been isolated from polysomes of mouse parotid glands. Filtration on Millipore and binding to poly(u) immobilized on Sepharose-4B, together with significant RNase-resistance (11-16%) indicated that this polysomal fraction contains large tracts of polyadenylate sequences. A single injection of isoproterenol, a procedure that causes, after a lag period of 20 hrs, a marked stimulation of DNA synthesis and cell division in salivary glands of rodents, induces a biphasic increase in the incorporation of (3H)-adenosine into poly(A)-associated polysomal RNA. The incorporation of (3H)-adenosine is already increased within 1 hour after stimulation, decreases at 3 hrs and increases again at 8 hrs, parallelling previously reported changes on chromatin template activity of mouse parotids. However, the percent poly(A) content, and the size of the poly(A)-associated RNA remain constant. Actinomycin D given 30 min prior to isoproterenol, inhibits isoproterenol-Stimulated poly(A)-rich RNA synthesis. These results demonstrate that, as part of the early molecular events of its action, isoproterenol regulates transcription of adenylate-rich RNA in mouse parotid gland.


Simulation assisted design for microneedle manufacturing: Computational modeling of two-photon templated electrodeposition.
Fully metallic micrometer-scale 3D architectures can be fabricated via a hybrid additive methodology combining multi-photon lithography with electrochemical deposition of metals. The methodology - referred to as two-photon templated electrodeposition (2PTE) - has significant design freedom that enables the creation of complicated, traditionally difficult-to-make, high aspect ratio metallic structures such as microneedles. These complicated geometries, combined with their fully metallic nature, can enable precision surgical applications such as inner ear drug delivery or fluid sampling. However, the process involves electrochemical deposition of metals into complicated 3D lithography patterns thicker than 500 μm. This causes potential and chemical gradients to develop within the 3D template, creating limitations to what can be designed. These limitations can be explored, understood, and overcome via numerical modeling. Herein we introduce a numerical model as a design tool that can predict growth for manufacturing complicated 3D metallic geometries. The model is successful in predicting the geometric result of 2PTE, and enables extraction of insights about geometric constraints through exploration of its mechanics.