Intravascular ultrasound evaluating coronary stents for patients with coronary artery disease: compared old with new multilink stents.
It was suggested that coronary stent design and coating may affect stent performance and hence induce varying degrees of thrombogenesis and neointimal hyperplasia. The purpose of this study is to compare the 6-month follow-up results between old and new Multilink stents with the method of intravascular ultrasound (IVUS) imaging. We have performed old (n = 40) and new (n = 35) Multilink stent implantations on 75 patients with coronary artery disease. Coronary angiography was performed before, immediately after, and 6 months after the in-stent procedure respectively. Six-month follow-up IVUS imaging was performed and analyzed off-line. Minimal lumen cross sectional area (CSA) of new Multilink stents was significantly larger than that of old Multilink stents (P = 0.0053). Mean stent lumen area of new Multilink stents was significantly larger than that of old Multilink stents (P = 0.040). Similarly, minimal lumen diameter (MLD) of new Multilink stents was larger than that of old Multilink stents (P = 0.011). Old Multilink stents had a higher percentage of plaque area than new Multilink stents. The new Multilink stent is obviously superior to old Multilink stents, in particular, in the stent MLD and lumen CSA--major determinants of the restenosis.


Ectopical expression of human MUC18 increases metastasis of human prostate cancer cells.
MUC18, a cell adhesion molecule (CAM), has been reported to be a diagnostic marker for the early detection of the metastatic potential of prostate cancers as well as implicated to be an important determinant for mediating the tumorigenesis and metastasis of prostate cancer. To test the hypothesis, we further investigated the possible role of MUC18 in the malignant progression of human prostate cancer. The human MUC18-minus, non-metastatic human prostate cancer LNCaP cells were transfected with the human cytomegalovirus immediate-early gene (HCMV-IE) promoter-driven human MUC18 (huMUC18) cDNA. The G418-resistant (G418R)-LNCaP clones that expressed a high level of huMUC18 were selected and used for testing the effect of huMUC18 expression on the in vitro growth, motility, and invasiveness as well as on the in vivo metastasis (via orthotopical injection) in a xenograft nude mouse model. HuMUC18 expression increased by four- to fivefold of in vitro motility and invasiveness of LNCaP cells. Anti-huMUC18 antibody significantly inhibited the in vitro motility and invasiveness of huMUC18-expressing LNCaP clones, but not the control clones. We suggest that huMUC18 expression is responsible for increasing these behaviors of LNCaP cells. HuMUC18 expression also directly increased the in vivo metastatic abilities of the LNCaP cells from the prostate gland to multiple distant organs. Western blot and immunohistochemistry analyses showed that the prostatic tumors as well as metastatic lesions expressed high levels of MUC18, indicating that they originated from the injected huMUC18-expressing LNCaP cells. We therefore conclude that HuMUC18 is an important determinant in increasing metastasis of human prostate cancer LNCaP cells to distant organs in a nude mouse model.


Clinical - pathological significance of leptin receptor (LEPR) expression in squamous cell carcinoma of the skin.
Adipokine leptin functions through its transmembrane receptors (LEPR). In many malignant tumors it stimulates the growth, migration and invasion of malignant cells. The aim of our work is to examine the effect of LEPR expression on the clinical-morphological properties of squamous cell carcinoma of the skin (cSCC). The biopsy material obtained by excision of squamous cell skin cancer was used. The test group consisted of excision biopsies of squamous cell carcinoma of the skin (n = 62), and the control group (n = 62) consisted of excision biopsies of non-tumor tissue of the skin (from the tumor environment) from an operative preparation delivered to the Pathohistology Department. After routine processing and paraffin molding, histochemical Hematoxylin-Eosin and immunohistochemical ABC method with anti LEPR and Ki67 antibodies were applied at 4 μm sections. The statistical software package SPSS for Windows (26.0) was used to analyze obtained results. Intracytoplasmic and intramembranous LEPR expression was found in 100 % of examined cSCCs. LEPR expression was statistically significantly associated with proliferation index and histologic grade of tumors. Pronounced LEPR expression was associated with a high proliferation index in 66.7 % of cases and with poorly differentiated cSCC in 94.4 %. Multivariate regression analysis showed that cSCCs with pronounced LEPR expression were seven times more often poorly differentiated than tumors with moderate or LEPR expression in trace. Our results indicate that LEPR expression is a predictor of the malignant potential of cSCC, so that based on LEPR expression, it is possible to identify an aggressive cSCC phenotype, which provides the possibility of individualizing anti-tumor treatment using LEPR antagonists.


Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest.
For decoding the mechanism of how cells and organs function information on their ultrastructure is essential. High-resolution 3D imaging has revolutionized morphology. Serial block face scanning electron microscopy (SBF-SEM) offers non-laborious, automated imaging in 3D of up to ~ 1 mm3 large biological objects at nanometer-scale resolution. For many samples there are obstacles. Quality imaging is often hampered by charging effects, which originate in the nonconductive resin used for embedding. Especially, if the imaged region of interest (ROI) includes the surface of the sample and neighbours the empty resin, which insulates the object. This extra resin also obscures the sample's morphology, thus making navigation to the ROI difficult. Using the example of small arthropods and a fish roe we describe a workflow to prepare samples for SBF-SEM using the minimal resin (MR) embedding method. We show that for imaging of surface structures this simple approach conveniently tackles and solves both of the two major problems-charging and ROI localization-that complicate imaging of SBF-SEM samples embedded in an excess of overlying resin. As the surface ROI is not masked by the resin, samples can be precisely trimmed before they are placed into the imaging chamber. The initial approaching step is fast and easy. No extra trimming inside the microscope is necessary. Importantly, charging is absent or greatly reduced meaning that imaging can be accomplished under good vacuum conditions, typically at the optimal high vacuum. This leads to better resolution, better signal to noise ratio, and faster image acquisition. In MR embedded samples charging is minimized and ROI easily targeted. MR embedding does not require any special equipment or skills. It saves effort, microscope time and eventually leads to high quality data. Studies on surface-linked ROIs, or any samples normally surrounded by the excess of resin, would benefit from adopting the technique.


Synthesis of the 6-deoxytalose-containing tetrasaccharide of the glycopeptidolipid from Mycobacterium intracellare serotype 7.
An efficient synthesis of 4-methoxyphenyl alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->2)-6-deoxy-alpha-L-Talp, the tetrasaccharide related to the GPLs of Mycobacterium intracellare serotype 7, was achieved with 4-methoxyphenyl 3,4-di-O-benzoyl-6-deoxy-alpha-L-talopyranoside (6c) as the key intermediate which was obtained through selective 3-O-benzoylation of 4-O-benzoyl-6-deoxy-alpha-L-taloside. Coupling of 6c with 3-O-allyloxycarbonyl-2,4-di-O-benzoyl-alpha-L-rhamnopyranosyl trichloroacetimidate followed by removal of the allyloxycarbonyl protecting group afforded the disaccharide acceptor 11. Condensation of 11 with 2,3,4-tri-O-benzoyl-alpha-L-rhamnopyranosyl-(1-->3)-2,4-di-O-benzoyl-alpha-L-rhamnopyranosyl trichloroacetimidate and subsequent deprotection gave the target tetrasaccharide.


Different ways to approach shale reservoirs' CO2 storage potential in America.
To achieve the Net Zero Carbon Emissions (NZCE) target by 2050, Carbon Capture, Utilization, and Storage (CCUS) is a major method. Gathering and injecting CO2 into shale reservoirs is an effective way to reduce the CO2 amount in the air and thus, release the greenhouse effect. CO2 injection into organic-rich shales could provide dual benefits of incremental oil or gas recovery and secure CO2 storage. When planning the CO2 injection project, the most important question is how much gas could be stored in the reservoir. Methods for calculating CO2 storage potential in shale reservoirs have been studied by many researchers. However, few researchers put those methods together and make comparisons to each other. This paper summarized five methods for evaluating CO2 storage potential in five shale reservoirs by using the literature published in recent years. This paper aims to discuss and evaluate the technical aspects related to gas storage. Those geomechanical properties, petrophysical properties, and construction parameters were discussed. Among those parameters, CO2 injection rate, skin factor, and Knudsen diffusion could significantly affect CO2 storage potential evaluation results. Also, if well integrity, especially cement quality, and permeability could be taken into consideration, CO2 storage simulation models' results will be more realistic. The significances of this study are: (1) served as guidance in calculating CO2 storage capacity in shale oil plays; (2) provides analyses in evaluating nowadays methods' limitations; (3) gives recommendations to researchers on how to improve those methods or create a new one.


Intravitreal bevacizumab for choroidal neovascularization in ocular histoplasmosis.
To define the role of intravitreal bevacizumab in individuals with choroidal neovascularization (CNV) resulting from Ocular Histoplasmosis syndrome (OHS). Retrospective chart review of a surgical therapy. We reviewed the course of 28 eyes of 28 patients who underwent intravitreal injection of bevacizumab for treatment of CNV secondary to OHS. Outcome was measured by pretreatment and posttreatment visual acuity (VA). The average pretreatment logarithm of the minimum angle of resolution (logMAR) VA was 0.65 (Snellen equivalent of 20/88). Mean follow-up was 22.43 weeks with an average of 1.8 intravitreal injections. Average final logMAR VA was 0.43 (Snellen equivalent of 20/54). Twenty eyes (71%) experienced an increase in central VA, whereas four eyes (14%) were unchanged and four eyes (14%) experienced a decrease in vision. Intravitreal bevacizumab may improve or stabilize VA in a significant majority of patients with neovascular complications of OHS (24 eyes [85.7%] in our study population).


Cranial Investigations of Crested Porcupine (Hystrix cristata) by Anatomical Cross-Sections and Magnetic Resonance Imaging.
This paper aimed to describe an atlas of the crested porcupine (Hystrix cristata) head by applying advanced imaging techniques such as MRI. Furthermore, by combining the images acquired through these techniques with anatomical sections, we obtained an adequate description of the structures that form the CNS and associated structures of this species. This anatomical information could serve as a valuable diagnostic tool for the clinical evaluation of different pathological processes in porcupines, such as abscesses, skull malformations, fractures, and neoplasia.


Access to Cytotoxic Medicines by Children With Cancer: A Focus on Low and Middle Income Countries.
The Essential Medicines Working Group of the International Society of Pediatric Oncology (SIOP) has proposed a list of antineoplastic drugs that should be available in low and middle income countries. Data were extracted on the listing of 18 essential and 8 ancillary antineoplastic medicines in the national essential medicines lists (NEMLs) or national reimbursable medicines lists (NRMLs) of 135 countries with gross national income (GNI) per capita of less than US $25,000. Correlations between numbers of medicines listed and GNI per capita, annual government health expenditure (AGHE) per capita, and the number of physicians per million people were examined. Listing of the 18 essential antineoplastic drugs ranged from 27% (thioguanine) to 95% (methotrexate). The median number of medicines listed was 7 (0-18) in low income countries (n = 26) and 14 in lower-middle (n = 42), upper-middle (n = 44), and high income countries (n = 20). For the ancillary eight medicines, the median was one (0-8) across the 135 countries. Correlations with GNI per capita (r = 0.17, P = 0.0266) and physician density (r = 0.25, P = 0.0017) were statistically significant; not so for AGHE per capita (r = 0.00, P = 0.5000). There was large variability within income groups in numbers of antineoplastic agents identified as essential in NEMLs and NRMLs. While not a direct measure of availability, listing is an important step, guiding procurement for the public sector. These results focus attention on deficits in NEMLs and NMRLs as a step to improving access to effective antineoplastic medicines for cancers in children in low and middle income countries.


Replication of a microsatellite genome-wide association study of Behcet's disease in a Korean population.
Behçet's disease is one of the major aetiologies of uveitis causing blindness in Asian countries. A genome-wide association study identified six microsatellite markers as disease susceptibility loci for Japanese patients with Behçet's disease. To confirm our recent results, these microsatellite markers were examined in a Korean population as a replication study. Study participants included 119 Behçet's disease patients and 141 controls. All were enrolled in Korea. Association between the six reported microsatellite markers (D3S0186i, D6S0014i, D6S0032i, 536G12A, D12S0645i and D22S0104i) and Behçet's disease was analysed. HLA-B was genotyped by sequence-based typing methods. A microsatellite marker located near the HLA-B region demonstrated significant association with Behçet's disease (P = 0.028). The genotype and phenotype frequencies of the HLA-B*51 gene were significantly increased in patients (23.1 and 39.5%, respectively) compared with healthy controls (11.2 and 20.1%, respectively; P < 0.001). Microsatellite analysis revealed that the HLA-B*51 gene was strongly associated with Behçet's disease in a Korean population.


iTRAQ-based quantitative proteomic analysis of Gloeothece sp. PCC 6909: Comparison with its sheathless mutant and adaptations to nitrate deficiency and sulfur limitation.
Gloeothece sp. PCC 6909 is a unicellular N(2)-fixing cyanobacterium with a well defined and highly developed sheath surrounding its cells. A sheathless mutant of this strain was previously obtained by chemical mutagenesis and, although lacking the sheath, it releases large amounts of polysaccharides into the culture medium. To provide a global understanding on the metabolic differences between the two phenotypes, the proteomes of the wild type and mutant were analyzed using a cross-species proteomics approach coupled with iTRAQ isobaric tagging technology, since their genome sequences are not yet available. Effects arising from the presence/absence of nitrate and sulfur are presented as two metabolically directed follow-up iTRAQ studies. These nutrients are believed to play a major role in Gloeothece's metabolism, including the production of extracellular polymeric substances - EPS. 454, 124, and 53 proteins were identified and reliably quantified using homology anchoring approaches for iTRAQ previously described. The results obtained strongly suggest that the chemical mutagenesis affected the regulation of a number of key cellular processes, as revealed by the significant fold changes observed for proteins covering a large spectrum of functional groups. Moreover, they provide new insights on the adaptations of Gloeothece cells to nitrate-deficiency and sulfur-limitation.


Understanding evaporation characteristics of a drop of distilled sulfur mustard (HD) chemical agent from stainless steel and aluminum substrates.
We report herein the evaporation rates and mechanism of a drop of distilled sulfur mustard (HD) agent from stainless steel and aluminum substrates. For systematic analysis, we used a laboratory-sized wind tunnel, thermal desorption (TD) connected to gas chromatograph/mass spectrometry (GC/MS) and drop shape analysis (DSA). We found that the evaporation rates of HD from stainless steel and aluminum increased with temperature. The rates were also linearly proportional to drop size. The time-dependent contact angle measurement showed that the evaporation of the drop of HD proceeded only by constant contact area mechanism from stainless steel surface. On the other hand, the evaporation of HD from aluminum proceeded by a combined mechanism of constant contact area mode and constant contact angle mode. Our experimental data sets and analysis could be used to predict vapor and contact hazard persistence of chemical warfare agents (CWAs) in the air and on exterior surfaces with chemical releases, which assists the military decision influencing personnel safety and decontamination of the site upon a chemical attack event.


Oxiapoptophagy: A type of cell death induced by some oxysterols.
Oxysterols are oxidized forms of cholesterol generated from cholesterol by auto-oxidation, enzymatic processes, or both. Some of them (7-ketocholesterol, 7β-hydroxycholesterol and 24(S)-hydroxycholesterol), when used at cytotoxic concentrations on different cell types from different species (mesenchymal bone marrow cells, monocytic cells and nerve cells), induce a type of cell death associated with OXIdative stress and several characteristics of APOPTOsis and autoPHAGY, defined as oxiapoptophagy. Oxidative stress is associated with overproduction of ROS, increased antioxidant enzyme activities, lipid peroxidation and protein carbonylation. Apoptosis is associated with activation of the mitochondrial pathway, opening of the mitochondrial permeability pore, loss of mitochondrial membrane potential, caspase-3 activation, PARP degradation, nuclear condensation and/or fragmentation. Autophagy is characterized by autophagic vacuoles revealed by monodansylcadaverine staining and transmission electron microscopy, plus increased ratio of LC-3II/LC-3I. In addition, morphological, topographical and functional changes of the peroxisome are observed. LINKED ARTICLES: This article is part of a themed issue on Oxysterols, Lifelong Health and Therapeutics. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v178.16/issuetoc.


A novel defensin-like antimicrobial peptide from the skin secretions of the tree frog, Theloderma kwangsiensis.
Defensins are one of the major families of antimicrobial peptides (AMPs), and have been reported from prokaryotic to eukaryotic kingdoms. But defensins are seldom found in amphibian which is a major resource of novel AMPs. A novel defensin-like AMP (defensin-TK) was isolated and characterized from skin secretions of the tree frog Theloderma kwangsiensis. The cDNA encoding defensin-TK precursor was cloned from the skin cDNA library of T. kwangsiensis. The deduced precursor of defensin-TK was composed of three domains, a signal peptide of 16 residues, a spacer peptide of 1 residues and a mature peptide of 42 residues. The mature peptide of defensin-TK shared the highest identity with the salamander (Cynops fudingensis) defensin CFBD-1. The six conserved cysteines which formed intramolecular disulfide bonds of defensins also exist in defensin-TK. Phylogenetic analysis indicated that defensin-TK was closely related to fish β-defensins. Defensin-TK showed potent and broad-spectrum antimicrobial activity. In addition, defensin-TK exerted a low hemolytic activity on human red cells. These results suggested defensin-TK might play an important role in defense the skin pathogenic microbes of tree frog T. kwangsiensis, and might be a promising candidate for development of novel antimicrobial agents.


Perfluoroalkyl substances (PFASs) decrease the expression of recombination-activating genes (RAG1 and RAG2) in human B lymphoma Namalwa cells.
Per- and polyfluoroalkyl substances (PFASs) are omnipresent and have been shown to induce a wide range of adverse effects, including hepatotoxicity, developmental toxicity and immunotoxicity. So far, little information is available about the mechanisms underlying the toxicity of PFASs, including those related to their immunotoxicity. Reported immunotoxic effects of PFASs include decreased antibody responses in experimental animals and humans, indicating that PFASs may, among others, affect B cell function. In the present study, we first assessed the effects of PFOA on the transcriptome of the human Namalwa B cell line using RNA seq analysis. Gene expression changes, analyzed using Ingenuity Pathway Analysis, pointed to various cellular processes affected by PFOA, including 'B cell development' and 'Primary immunodeficiency signaling'. Interestingly, PFOA decreased the expression of RAG1 and RAG2, genes involved in immunoglobulin and T cell receptor V(D)J recombination. As a next step, time- and concentration-dependent changes in the expression of RAG1 and RAG2 upon exposure to PFOA, PFNA, PFHxS and PFOS were studied through RT-qPCR analysis. Analysis with the concentration-response modeling software PROAST resulted in the following potency ranking: PFNA > PFOA > PFOS > PFHxS. Altogether, the present in vitro study provides insights into the effects of selected PFASs on B cells, identifying RAG1 and RAG2 expression as possible relevant targets that may play a role in the immunotoxicity of PFASs.


Synthesis of some epoxy and/or N-oxy 17-picolyl and 17-picolinylidene-androst-5-ene derivatives and evaluation of their biological activity.
Steroidal epoxy and/or N-oxy 17-picolyl and 17-picolinylidene-androst-5-ene derivatives have been prepared using 3beta,17beta-dihydroxy-17alpha-picolyl-androst-5-ene (1), 3beta-acetoxy-17-picolinylidene-androst-5-ene (2), and 3beta-hydroxy-17-picolinylidene-androst-5-ene (3) as synthetic precursors. The compounds 2 and/or 3 were reacted with m-chloroperoxybenzoic acid (MCPBA). The compounds synthesized from 2 were 17-picolinylidene-N-oxide 4, 5alpha,6alpha-epoxy and 5beta,6beta-epoxy-17-picolinylidene-N-oxide 5 and 6, and 5alpha,6alpha:17alpha,20alpha- and 5beta,6beta:17alpha,20alpha-diepoxy-N-oxide 7 and 8. Starting from compound 3, a mixture of 5alpha,6alpha-epoxy and 5beta,6beta-epoxy-17-picolinylidene 9 and 10, 5alpha,6alpha-epoxy and 5beta,6beta-epoxy-17-picolinylidene-N-oxide 11 and 12, and 5alpha,6alpha:17alpha,20alpha- and 5beta,6beta:17alpha,20alpha-diepoxy-N-oxide 13 and 14 were obtained. From compounds 15 and 18, obtained from 1 and 3 by the Oppenauer oxidation, the 4alpha,5alpha-epoxy and 4beta,5beta-epoxy derivatives 16, 17 and 20, 21 were prepared by oxidation with 30% H(2)O(2). Oxidation of 18 with MCPBA yielded only the N-oxide 19. The structures of compounds 15 and 18 were proved by the X-ray analysis. Compounds 1-6, 9, 15, 17, 18, and 21 were tested on activity against the enzyme aromatase. Antitumor activity against three tumor cell lines (human breast adenocarcinoma ER+, MCF-7, human breast adenocarcinoma ER-, MDA-MB-231, and prostate cancer PC3) was evaluated. Three tested compounds (1, 4, and 19) showed strong activity against PC3, the IC(50) values being in the range 0.55-10microM, whereas compound 17 showed strong activity against MDA-MB-231 (IC(50) 10.4microM).


Results of a phase II study of thalidomide and azacitidine in patients with clinically advanced myelodysplastic syndromes (MDS), chronic myelomonocytic leukemia (CMML) and low blast count acute myeloid leukemia (AML).
Single agent azacitidine or immunomodulatory drugs are effective in myelodysplastic syndrome (MDS), with differing target mechanisms and toxicities. Objectives of this ALLG MDS3 study in clinically advanced MDS, AMML and low blast AML were to establish safety, response and quality of life of azacitidine and thalidomide. Patients received azacitidine (75mg/m2/d sc 7days every 28 days), and oral thalidomide up to 100mg/d for maximum 12months. Eighty patients registered; median age 68 years (range 42-82), 49% IPSS int2-high. With 36.5 months follow up, patients received median 9 cycles azacitidine, 6.1mths thalidomide. Nonhematologic toxicity grade 3+ in 85%, commonly infections. Overall response rate was 63%; 26% CR were unaffected by IPSS. Median response duration 26.3months; overall survival was 28.1months. This combination azacitidine and thalidomide in clinically advanced MDS, CMML and low-blast AML was tolerable without unexpected toxicity and encouraging responses support further investigation of combination approaches with hypomethylating agent and immunomodulatory drug.


Trans-amniotic stem cell therapy (TRASCET) minimizes Chiari-II malformation in experimental spina bifida.
We sought to study the impact of trans-amniotic stem cell therapy (TRASCET) in the Chiari-II malformation in experimental spina bifida. Sprague-Dawley fetuses (n=62) exposed to retinoic acid were divided into three groups at term (21-22 days gestation): untreated isolated spina bifida (n=21), isolated spina bifida treated with intra-amniotic injection of concentrated, syngeneic, labeled amniotic fluid mesenchymal stem cells (afMSCs) on gestational day 17 (n=28), and normal controls (n=13). Analyses included measurements of brainstem and cerebellar placement on high resolution MRI and histology. Statistical comparisons included ANOVA. In parallel to the expected induced coverage of the spina bifida in the afMSC-treated group (P<0.001), there were statistically significant differences in brainstem displacement across the groups (P<0.001), with the highest caudal displacement in the untreated group. Significant differences in cerebellar displacement were also noted, albeit less pronounced. Pairwise comparisons were statistically significant, with P=0.014 between treated and normal controls in caudal brainstem displacement and P<0.001 for all other comparisons. Labeled afMSCs were identified in 71% of treated fetuses. Induced coverage of spina bifida by TRASCET minimizes the Chiari-II malformation in the retinoic acid rodent model, further suggesting it as a practical alternative for the prenatal management of spina bifida.


In vivo effects of single or combined dietary antimutagens on mutagen-induced chromosomal aberrations.
The food components chlorophyllin, beta-carotene and alpha-linolenic acid (in its methyl ester form) were tested in Chinese hamsters for antimutagenic activity on the powerful mutagen thio-tepa. Each of these natural protective compounds inhibited by 70-85% the clastogenic effects induced by the mutagen. When 2 or 3 of these antimutagens were administered simultaneously no additive effects were observed. alpha-Linolenic acid methyl ester was the most effective antimutagen under the experimental conditions.


Adenocarcinomas of the oral cavity: a clinicopathologic study of terminal duct carcinomas.
A clinico-pathologic study of 12 patients, each harboring a hitherto not delineated adenocarcinoma of salivary origin is presented. The authors have designated this histologically unique carcinoma as 'terminal duct adenocarcinoma' in deference not only to its light-optic appearance, but also to a putative origin from the reserve cells (epithelial and myoepithelial) of the intercalated duct. The tumors' local invasive properties with extension into nerves and adjacent bone suggest their biologic behavior is like that of adenoid cystic carcinomas.


Atherogenic diet-diminished endothelial glycocalyx contributes to impaired vasomotor properties in rat.
Hypercholesterolemia- and atherosclerosis-caused vasomotor property dysfunction may be involved in many clinic manifestations of atherosclerosis, including angina, acute myocardial infarction, and sudden cardiac death. However, its underlying mechanism is not clear. The endothelial glycocalyx is a protective surface layer on the endothelial cells, serving as a molecular sieve, cell adhesion modulator, and mechanosensor for blood flow. In the present study, we demonstrated by confocal microscopy in Sprague-Dawley (SD) male rats fed a 12-wk high-cholesterol diet (HC) compared with the normal diet (NC) that the dimension of the endothelial glycocalyx reduced significantly in both the common carotid artery (2.89 ± 0.41 µm and 3.25 ± 0.44 μm, respectively) and the internal sinus region (2.35 ± 0.07 µm and 3.46 ± 0.86 μm, respectively). Furthermore, we showed by real-time PCR that this dimension modification of endothelial glycocalyx may be attributed to a significant downregulation of heparan sulfate proteoglycan (HSPG)-related genes, including syndecan-3, glypican-1, and EXT1, not resulting from an enhanced shedding of sulfated glycosaminoglycans (sGAGs) from the vessel wall to the plasma. Meanwhile, the mean contraction and relaxation forces of the common carotid artery with responses to norepinephrine (NE) and acetylcholine (ACh) decreased ~0.34- and 0.13-fold, respectively, accompanied by a lower level of nitric oxide (NO) release. These findings suggest that the atherogenic high cholesterol diet diminished endothelial glycocalyx and disturbed the local NO release, thus contributing to the impaired vasomotor properties of the vessel.NEW & NOTEWORTHY Twelve-week high-cholesterol (HC) diet reduces the thickness of the endothelial glycocalyx in Sprague-Dawley (SD) male rats, which is mainly attributed to a downregulation of heparan sulfate proteoglycan-related genes (syndecan-3, glypican-1, EXT1), not resulting from an enhanced shedding of sulfated glycosaminoglycans (sGAGs) into the plasma. HC-diminished glycocalyx may disturb its mechanotransduction of local shear stress, lower nitric oxide (NO) release, and impair vasomotor responses to norepinephrine (NE) and acetylcholine (ACh).


Medication non-adherence and therapeutic inertia independently contribute to poor disease control for cardiometabolic diseases.
Poorly controlled cardiometabolic biometric health gap measures [e.g.,uncontrolled blood pressure (BP), HbA1c, and low-density lipoprotein cholesterol (LDL-C)] are mediated by medication adherence and clinician-level therapeutic inertia (TI). The study of comparing relative contribution of these two factors to disease control is lacking. We conducted a retrospective cohort study using 7 years of longitudinal electronic health records (EHR) from primary care cardiometabolic patients who were 35 years or older. Cox-regression modeling was applied to estimate how baseline proportion of days covered (PDC) and TI were associated with cardiometabolic related health gap closure. 92,766 patients were included in the analysis, among which 89.9%, 85.8%, and 73.3% closed a BP, HbA1c, or LDL-C gap, respectively, with median days to gap closure ranging from 223 to 408 days. Patients who did not retrieve a medication were the least likely to achieve biometric control, particularly for LDL-C (HR = 0.58, 95% CI: 0.55-0.60). TI or uncertainty of TI was associated with a high risk of health gap persistence, particularly for LDL-C (HR ranges 0.46-0.48). Both poor medication adherence and TI are independently associated with persistent health gaps, and TI has a much higher impact on disease control compared to medication adherence, implying disease management strategies should prioritize reducing TI.


HnRNP A2/B1 as a potential anti-tumor target for triptolide based on a simplified thermal proteome profiling method using XGBoost.
Triptolide (TP) is a highly active natural medicinal ingredient with significant potential in anticancer. The strong cytotoxicity of this compound suggests that it may have a wide range of targets within cells. However, further target screening is required at this stage. Traditional drug target screening methods can be significantly optimized using artificial intelligence (AI). This study aimed to identify the direct protein targets and explain the multitarget action mechanism of the anti-tumor effect of TP with the help of AI. The CCK8, scratch test, and flow cytometry analysis were used to examine cell proliferation, migration, cell cycle, and apoptosis in tumor cells treated with TP in vitro. The anti-tumor effect of TP in vivo was evaluated by constructing a tumor model in nude mice. Furthermore, we established a simplified thermal proteome analysis (TPP) method based on XGBoost (X-TPP) to rapidly screen the direct targets of TP. We validated the effects of TP on protein targets through RNA immunoprecipitation and pathways by qPCR and Western blotting. TP significantly inhibited tumor cell proliferation and migration and promoted apoptosis in vitro. Continuous administration of TP to tumor mice can significantly suppress tumor tissue size. We verified that TP can affect the thermal stability of HnRNP A2/B1 and exert anti-tumor effects by inhibiting HnRNP A2/B1-PI3K-AKT pathway. Adding siRNA to silence HnRNP A2/B1 also significantly down-regulated expression of AKT and PI3K. The X-TPP method was used to show that TP regulates tumor cell activity through its potential interaction with HnRNP A2/B1.


Artifacting Identity. How Grillz, Ball Gags and Gas Masks Expand the Face.
By questioning the attribution of a primary role to the eyes as bearers of identity within traditional Western culture, this paper will problematize the agentivity performed by the lower mereology of the face, identified with the mouth-nose assemblage. In particular, the study will focus on the manipulation of such facial spatiality through the intervention of three "lower face" artifacts: the grill, the ball gag and the gas mask. This piece of work will examine their plastic and figurative dimensions in the technological interaction with the facial organs. Furthermore, we will take into consideration the sociocultural context of wearability performed by the different bearers with the aim of grasping the identity shift that the artifacts trigger. The study, therefore, will organize the corpus as a sequence that starts inside the oral cavity where the grill is worn; then moves to a progressive exteriority with the ball gag that emerges from the mouth through the straps fastened around the head; eventually dealing with the exterior projection operated by the gas mask which by means of its filters portends beyond the anatomical face. Ultimately the three artifacts are presented as a threefold articulation of a liminal agency towards an expanded form of humanity including animality embedded within and without the space of meaning represented by the face.


Full-face photorejuvenation of photodamaged skin by intense pulsed light with integrated contact cooling: initial experiences in Asian patients.
For Asian skin, recent "non-ablative" skin rejuvenation techniques have become the focus of attention for darker complected patients. In our earlier research, we have shown that intense pulsed light (IPL) technology can be applied to Asian skin with a high degree of safety and efficacy. In this study, we performed full-face photorejuvenation using a new IPL device incorporating a 560 nm filter and integrated contact cooling system for the improvement of various symptoms associated with photoaging in Asian patients. A total of 73 patients were treated with a series of five or more full-face treatments at 3-4 week intervals using IPL with integrated contact cooling. One month after the third and fifth treatments, the patient and the treating physicians subjectively evaluated improvement in five areas-in pigmentation, telangiectasia, fine wrinkles, skin texture, and over-all improvement. In addition, histological changes were evaluated. Pigmentation improvement, telangiectasia reduction, fine wrinkle reduction, smoother skin texture, and over-all improvement were evaluated according to five grades of percentage improvement. In addition, the subjective rating by the patients and the physicians was averaged, and the combined results were evaluated. After the fifth treatment, a combined rating of greater than 60% improvement was given to more than 80% of patients for pigmentation improvement, telangiectasia reduction or removal, smoother skin texture, and overall improvement. Histological evaluations showed strong staining of Type I and Type III collagen. Complications were minor and transitory, with burning sensations and erythema in only two patients. Full-face photorejuvenation for Asian patients using this device is not only effective but is also associated with fewer post-treatment complications than other more invasive modalities. We conclude that IPL photorejuvenation can be the basis for safe and effective skin rejuvenation in Asian patients.


Empirical and bioinformatic characterization of buffalo (Bubalus bubalis) colostrum whey peptides & their angiotensin I-converting enzyme inhibition.
Whey based peptides are well known for their nutritional and multifunctional properties. In this context, whey proteins from buffalo colostrum & milk were digested by in vitro simulation digestion and analyzed by nano-LC-MS/MS. Functional protein association networks, gene annotations and localization of identified proteins were carried out. An ACE inhibitory peptide sorted from the library was custom synthesized and an in vitro ACE assay was performed. The study led to the identification of 74 small peptides which were clustered into 5 gene functional groups and majority of them were secretory proteins. Among the identified peptides, majority of them were found identical to angiotensin I-converting enzyme (ACE) inhibitors, antioxidant, antimicrobial, immunomodulatory and opioidal peptides. An octapeptide (m/z - 902.51, IQKVAGTW) synthesized was found to inhibit ACE with an IC50 of 300±2µM. The present investigation thus establishes newer vista for food derived peptides having ACE inhibitory potential for nutraceutical or therapeutic applications.


Forming and immunological properties of some lipopolysaccharide-chitosan complexes.
The complex formation of lipopolysaccharide (LPS) with chitosan (Ch) was demonstrated using sedimentation velocity analysis in the analytical ultracentrifuge, centrifugation in glycerol gradient and isopicnic centrifugation in cesium chloride. An addition of Ch to the Escherichia coli and Yersinia pseudotuberculosis LPS solutions was found to result in formation of the stable LPS-Ch complexes. The interaction is a complicated process and depends on time and reaction temperature, as well as on the molecular weight of chitosan. A stable LPS-Ch complex could be formed only after preliminary incubation of the initial components at an elevated temperature (37 degrees C). It should be noted that process of LPS complexation with Ch is accompanied by additional dissociating of LPS. The complex formation was shown to be a result not only of ionic binding, but also of other types of interactions. The interaction of Ch with LPS was shown to modulate significantly the biological activity of LPS. The LPS-Ch complex (1:5 w/w) was shown to possess much lower toxicity in a comparison with the parent LPS at injection to mice in the similar concentration. The LPS-Ch complex was shown to maintain an ability to induce of IL-8 and TNF, but induction of IL-8 and TNF biosynthesis by the LPS-Ch complex was lower than that by the parent LPS. The complex LPS-Ch, similarly to the parent LPS, was found stimulated the formation of the IL-8 in the dose-dependent manner in the human embryonal kidney cells (HEK 293 cells) transfected with TLR4 in combination with MD2.


Biological activities of cyclic enkephalin pseudopeptides containing thioamides as amide bond replacements.
Analogs of H-Tyr-cyclo(N epsilon-D-Lys-Gly-Phe-Leu) have been prepared which contain thioamides at the 3-4 position (monothio), 3-4 and 5-2 positions (dithio), and 2-3, 3-4, and 5-2 positions (trithio). These compounds have been tested for opioid activity in mu- and delta-receptor selective bio- and binding assays. As the number of sulfurs increased, the biological activities dropped on the guinea pig ileum and fluctuated modestly on the mouse vas deferens assay. Surprisingly, the compounds displayed increasing delta selectivity as the number of sulfurs increased. In the binding assay, the thioamide analogs tended to retain affinity toward the mu receptor. The mono- and dithio-analogs were more mu selective than the parent, while the trithio-analog was more delta selective. These results suggest that the subtle exchange of sulfur for oxygen can have a significant impact on receptor selectivity and affinity, and probably reflect the different conformation/structural requirements for binding vs. the biological transduction event.


A method for gel grade determination and application evaluation of two citrus pectins.
Citrus paradisi Macf. cv. Changshanhuyou and Citrus paradisi Macf. cv. Star Ruby are two emerging processed citrus fruits. The processing produces lots of peel wastes rich in pectin. While more attentions were paid on pectin's functional properties, the quality about commercial application like gel grade was little investigated. In this study, we established a method for gel grade determination based on texture analyzer, the new method is economical and can be used on a large scale in the laboratory. The commercial application related qualities of two citrus pectins were also studied in detail. The results showed that the yields of Changshanhuyou and Star Ruby pectins (CHP and SRP) were 20.23 % and 18.33 %, respectively. The indexes of CHP and SRP mostly were in line with the commodity standards, except the dry weight loss. The gel grades of CHP and SRP determined by the new method were 109.9 and 96.8, respectively. The CHP aqueous solution exhibited higher apparent viscosity and better performance in stabilizing acidified milk drink (AMD) compared with commercial pectin. From the view of commercial application related qualities and functional properties, CHP could be a good potential commercial pectin.


Expansion of the genetic code via expansion of the genetic alphabet.
Current methods to expand the genetic code enable site-specific incorporation of non-canonical amino acids (ncAAs) into proteins in eukaryotic and prokaryotic cells. However, current methods are limited by the number of codons possible, their orthogonality, and possibly their effects on protein synthesis and folding. An alternative approach relies on unnatural base pairs to create a virtually unlimited number of genuinely new codons that are efficiently translated and highly orthogonal because they direct ncAA incorporation using forces other than the complementary hydrogen bonds employed by their natural counterparts. This review outlines progress and achievements made towards developing a functional unnatural base pair and its use to generate semi-synthetic organisms with an expanded genetic alphabet that serves as the basis of an expanded genetic code.


Nox1-derived oxidative stress as a common pathogenic link between obesity and hyperoxaluria-related kidney injury.
Specific relationships among reactive oxygen species, activation pathways, and inflammatory mechanisms involved in kidney injury were assessed in a combined model of obesity and hyperoxaluria. Male Wistar rats were divided into four groups: Control, HFD (high fat diet), OX (0.75% ethylene glycol), and HFD + OX (combined model) Changes in basal O2- levels were evaluated by chemiluminescence in renal interlobar arteries and renal cortex. Furthermore, the effect of different inhibitors on NADPH-stimulated O2- generation was assessed in renal cortex. Oxidative stress sources, and local inflammatory mediators, were also determined, in parallel, by RT-PCR, and correlated with measures of renal function, urinary biochemistry, and renal structure. Rats from the HFD group developed overweight without lipid profile alteration. Tubular deposits of crystals were seen in OX and severely enhanced in HFD + OX groups along with a significantly higher impairment of renal function. Basal oxidative stress was increased in renal cortex of OX rats and in renal arteries of HFD rats, while animals from the combined HFD + OX group exhibited the highest levels of oxidative stress in renal cortex, derived from xanthine oxidase and COX-2. NADPH oxidase-dependent O2- generation was elevated in renal cortex of the OX group and markedly enhanced in the HFD + OX rats, and associated to an up-regulation of Nox1 and a down-regulation of Nox4 expression. High levels of oxidative stress in the kidney, of OX and HFD + OX groups were also associated to an inflammatory response mediated by an elevation of TNFα, COX-2, NFκB1 MCP-1, and OPN. Oxidative stress is a key pathogenic factor in renal disease associated to hyperoxaluria and a common link underlying the exacerbated inflammatory response and kidney injury found under conditions of both obesity and hyperoxaluria. Nox1 pathway must be considered as a potential therapeutic target.


Does pressure antagonize anesthesia? High-pressure stopped-flow study of firefly luciferase and anatomy of initial flash.
The antagonizing effect of high pressure against anesthesia is well known. With purified firefly luciferase, however,. Biophys. J. 60:1309-1314) reported that high pressure did not affect the initial flash intensity. Firefly luciferase emits a burst of light when the substrates luciferin and ATP are added in the presence of O2. The light intensity decays rapidly and the weak light lasts for hours. The initial flash is a transient event and is not in a steady state. The steady state is represented by the slope of the linear part of the integral of the light output. The present study used a high-pressure stopped-flow system to compare the pressure effects on the initial flash intensity and the steady-state light intensity. The flash intensity did not change by the application of hydrostatic pressure in the presence or absence of chloroform or 1-octanol. In contrast, high pressure increased the steady-state light intensity. The application of 12 MPa pressure increased the steady-state light intensity of firefly luciferase inhibited by 5 mM chloroform or 0.7 mM 1-octanol by 19.7% and 18.8%, respectively. When analyzed by the rapid reaction kinetics of the transition state theory, the initial peak intensity represents the total amount of active enzyme and is unrelated to the reaction rate. Anesthetics inhibited the initial flash by unfolding the protein, thereby decreasing the concentration of the active enzyme. Pressure affected the steady-state light intensity by changing the reaction rates.


Mixture model approach to tumor classification based on pharmacokinetic measures of tumor permeability.
To categorize the disease severity of mammary tumors in an animal model through the application of a novel tumor permeability mixture model within a hierarchical modeling framework. Thirty-six rats with mammary tumors of varying grade were imaged via dynamic contrast-enhanced (CE) MRI using albumin-(Gd-DTPA)30. Time-dependent contrast agent concentration curves for blood and tumor tissue were obtained and a mathematical model of microvascular blood-tissue exchange was developed under the hypothesis that endothelial integrity is disrupted in a manner proportional to the degree of malignancy, with benign tumors showing no disruption of the vasculature endothelium. This permeability model was incorporated into a statistical model for the benign and malignant tumor subgroups that enabled automatic subject classification. The structural and statistical models were implemented using the software Nonlinear Mixed Effects Modeling (NONMEM) to statistically separate subjects into the two subgroups. Individual tumor classifications (as benign or malignant) were evaluated against the Scarff-Bloom-Richardson microscopic scoring method as applied to the tumor histology of each subject. The model-based classification resulted in 90.9% sensitivity, 92.9% specificity, and 91.7% accuracy. Mixture model analysis provides a robust method for subject classification without user intervention and bias. Although the present results are promising, additional research is needed to further evaluate this technique for diagnostic purposes.


The 17-Gene Genomic Prostate Score Assay Is Prognostic for Biochemical Failure in Men With Localized Prostate Cancer After Radiation Therapy at a Community Cancer Center.
The objective of this study was to assess the association between the Oncotype DX Genomic Prostate Score (GPS) assay and long-term outcomes in men with localized prostate cancer (PCa) after radiation therapy (RT). We hypothesized that the GPS assay is prognostic for biochemical failure (BCF), along with distant metastasis (DM) and PCa-specific mortality in patients with PCa receiving RT. We retrospectively studied men with localized PCa treated with definitive RT at Georgia Urology from 2010 to 2016. The primary objective was to assess the association between GPS results and time to BCF per the Phoenix criteria; we also assessed time to DM and PCa-specific mortality. We used Cox proportional hazards regression models for all analyses, with clinicopathologic covariates determined a priori for multivariable modeling. A total of 450 patients (median age, 65 years; 35% Black) met eligibility criteria. There was a strong univariable association between GPS result and time to BCF (hazard ratio [HR] per 20-unit increase = 3.08; 95% confidence interval [CI], 2.11-4.46; P < .001), which persisted after adjusting for clinicopathologic characteristics in multivariable analyses. We also observed this association for time to DM (HR = 5.19; 95% CI, 3.06-8.77; P < .001) and PCa-specific mortality (HR = 13.07; 95% CI, 4.42-49.39; P < .001). Race was not a predictor of time to BCF or DM, and the GPS assay was strongly prognostic for all endpoints in Black and White patients. In a community-based cohort, the GPS assay was strongly prognostic for time to BCF as well as long-term outcomes in men treated with RT for localized PCa.


Strategies to regulate transcription factor-mediated gene positioning and interchromosomal clustering at the nuclear periphery.
In budding yeast, targeting of active genes to the nuclear pore complex (NPC) and interchromosomal clustering is mediated by transcription factor (TF) binding sites in the gene promoters. For example, the binding sites for the TFs Put3, Ste12, and Gcn4 are necessary and sufficient to promote positioning at the nuclear periphery and interchromosomal clustering. However, in all three cases, gene positioning and interchromosomal clustering are regulated. Under uninducing conditions, local recruitment of the Rpd3(L) histone deacetylase by transcriptional repressors blocks Put3 DNA binding. This is a general function of yeast repressors: 16 of 21 repressors blocked Put3-mediated subnuclear positioning; 11 of these required Rpd3. In contrast, Ste12-mediated gene positioning is regulated independently of DNA binding by mitogen-activated protein kinase phosphorylation of the Dig2 inhibitor, and Gcn4-dependent targeting is up-regulated by increasing Gcn4 protein levels. These different regulatory strategies provide either qualitative switch-like control or quantitative control of gene positioning over different time scales.


Simultaneous analysis of free and sulfo-conjugated steroid estrogens in artificial urine solution and agricultural soils by high-performance liquid chromatography.
A simple and robust analytical method was developed to simultaneously detect and quantify 17β-estradiol (E2), estrone (E1), 17β-estradiol-3-sulphate (E2-3S), and estrone-3-sulphate (E1-3S) in aqueous solutions (calcium chloride and artificial urine solutions) and agricultural soils using high performance liquid chromatography and UV detection. The standards for all four compounds were linear in the range of 0.01 to 1.0 μg mL(-1) (n = 6) and 1.0 to 20 μg mL(-1) (n = 6), respectively, with correlation coefficients > 0.999. The on-column limits of detection at an injection volume of 50 μL and S/N (signal: noise) ratio of 3 were: 9.0 ng mL(-1), 10 ng mL(-1), 5.0 ng mL(-1), and 7.0 ng mL(-1) for E2-3S, E1-3S, E2 and E1, respectively. The limit of detection and quantification in artificial urine solution and CaCl(2) solution was 1.0 ng mL(-1) for all four compounds. Method detection limits for the compounds in the 3 soils ranged from 2 to 2.4 ng g(-1) (E2-3S and E1-3S), and 1.0 to 2.9 ng g(-1) (E2 and E1), respectively.


[Variations of intraocular pressure induced by psychological stress (author's transl)].
Peculiar psychic reactivity of glaucomatous patients detected clinically and widely treated in the literature, has led us to study the behaviour of intraocular pressure in normal and glaucomatous subjects when submitted to various stresses. The data obtained show that prolonged stress increases intraocular pressure in normal subjects, unlike shorter stress which only causes an increase of intraocular pressure in glaucomatous patients. The possible genesis of the phenomenon is discussed.


Evaluation of the prevalence and factors associated with acute kidney injury in a pediatric intensive care unit.
To assess the prevalence of acute kidney injury in pediatric intensive care unit according to diagnostic criteria - pediatric risk, injury, failure, loss, end-stage renal disease, Acute Kidney Injury Network and Acute Kidney Injury Work Group, or Kidney Disease: Improving Global Outcomes -, and determining factors associated with acute kidney injury as well as its outcome. This was a cross-sectional monocentric observational study, including patients aged between 29 days and 17 years who were admitted to the pediatric intensive care unit between January 1, 2012 and December 31, 2016. To evaluate the association between the study variables and acute kidney injury, the log-binomial generalized univariate and multivariate linear models were adjusted. The study included 1131 patients, with prevalence of acute kidney injury according to the Acute Kidney Injury Network and Kidney Disease: Improving Global Outcomes criteria of 12.6% and of 12.9% according to the pediatric risk, injury, failure, loss, end-stage renal disease. In the multivariate analysis of older children (PR 1.007, 95% CI: 1.005-1.009), sepsis (PR 1.641, 95% CI: 1.128-2.387), demand for ventilatory support (PR 1.547, 95% CI: 1.095-2.186), and use of vasoactive amines (PR 2.298, 95% CI: 1.681-3.142) constituted factors associated with statistical significance to the development of acute kidney injury. The mortality rate among those with acute kidney injury was 28.7%. Older children, diagnosis of sepsis, demand for ventilatory support, and use of vasoactive amines were correlated with a higher risk of developing acute kidney injury. The mortality associated with acute kidney injury was elevated; it is crucial that all measures that ensure adequate renal perfusion are taken for patients with risk factors, to avoid the installation of the disease.


The action of physiological and synthetic steroids on the calcium channel CatSper in human sperm.
The sperm-specific channel CatSper (cation channel of sperm) controls the intracellular Ca2+ concentration ([Ca2+]i) and plays an essential role in sperm function. It is mainly activated by the steroid progesterone (P4) but is also promiscuously activated by a wide range of synthetic and physiological compounds. These compounds include diverse steroids whose action on the channel is so far still controversial. To investigate the effect of these compounds on CatSper and sperm function, we developed a high-throughput screening (HTS) assay to measure changes in [Ca2+]i in human sperm and screened 1,280 approved and off-patent drugs including 90 steroids from the Prestwick chemical library. More than half of the steroids tested (53%) induced an increase in [Ca2+]i and reduced the P4-induced Ca2+ influx in human sperm in a dose-dependent manner. Ten of the most potent steroids (activating and P4-inhibiting) were selected for a detailed analysis of their action on CatSper and their ability to act on sperm acrosome reaction (AR) and penetration in viscous media. We found that these steroids show an inhibitory effect on P4 but not on prostaglandin E1-induced CatSper activation, suggesting that they compete for the same binding site as P4. Pregnenolone, dydrogesterone, epiandrosterone, nandrolone, and dehydroepiandrosterone acetate (DHEA) were found to activate CatSper at physiologically relevant concentrations within the nanomolar range. Like P4, most tested steroids did not significantly affect the AR while stanozolol and estropipate slightly increased sperm penetration into viscous medium. Furthermore, using a hybrid approach integrating pharmacophore analysis and statistical modelling, we were able to screen in silico for steroids that can activate the channel and define the physicochemical and structural properties required for a steroid to exhibit agonist activity against CatSper. Overall, our results indicate that not only physiological but also synthetic steroids can modulate the activity of CatSper with varying potency and if bound to CatSper prior to P4, could impair the timely CatSper activation necessary for proper fertilization to occur.


Spectral Shape Recovery and Analysis Via Data-driven Connections.
We introduce a novel learning-based method to recover shapes from their Laplacian spectra, based on establishing and exploring connections in a learned latent space. The core of our approach consists in a cycle-consistent module that maps between a learned latent space and sequences of eigenvalues. This module provides an efficient and effective link between the shape geometry, encoded in a latent vector, and its Laplacian spectrum. Our proposed data-driven approach replaces the need for ad-hoc regularizers required by prior methods, while providing more accurate results at a fraction of the computational cost. Moreover, these latent space connections enable novel applications for both analyzing and controlling the spectral properties of deformable shapes, especially in the context of a shape collection. Our learning model and the associated analysis apply without modifications across different dimensions (2D and 3D shapes alike), representations (meshes, contours and point clouds), nature of the latent space (generated by an auto-encoder or a parametric model), as well as across different shape classes, and admits arbitrary resolution of the input spectrum without affecting complexity. The increased flexibility allows us to address notoriously difficult tasks in 3D vision and geometry processing within a unified framework, including shape generation from spectrum, latent space exploration and analysis, mesh super-resolution, shape exploration, style transfer, spectrum estimation for point clouds, segmentation transfer and non-rigid shape matching. The online version supplementary material available at 10.1007/s11263-021-01492-6.


IFN-γ‒Induced APOBEC3B Contributes to Merkel Cell Polyomavirus Genome Mutagenesis in Merkel Cell Carcinoma.
Merkel cell polyomavirus (MCPyV) is the causative agent of an aggressive skin tumor, Merkel cell carcinoma. The viral genome is integrated into the tumor genome and harbors nonsense mutations in the helicase domain of large T antigen. However, the molecular mechanisms by which the viral genome gains the tumor-specific mutations remain to be elucidated. Focusing on host cytosine deaminases APOBEC3s, we find that A3A, A3B, or A3G introduces A3-specific mutations into episomal MCPyV genomes in MCPyV-replicating 293-derivative cells. Sequence analysis of MCPyV genomes retrieved from the NCBI database revealed a decrease of TpC dinucleotide, a preferred target for A3A and A3B, in the 3'-region of the large T antigen‒coding sequence. The viral DNA isolated from tumors contained mutated cytosines, with a remarkable bias toward TpC dinucleotide. Analysis of publicly available microarray data showed that expression of IFN-γ and cytotoxic T lymphocyte markers was positively correlated with the A3A, A3B, and A3G levels in MCPyV-positive but not in MCPyV-negative tumors. Finally, IFN-γ treatment induced A3B and A3G expression in the MCPyV-positive Merkel cell carcinoma cell line MS-1. These results suggest that the IFN-γ-A3B axis plays pivotal roles in evolutionally shaping MCPyV genomic sequences and in generating tumor-specific large T antigen mutations during development of Merkel cell carcinoma.


Feasibility of a Cinematic-Virtual Reality Training Program for Health Professional Students: A Single-Arm Pre-Post Study.
Cinematic-virtual reality (cine-VR) has demonstrated improvements in cultural self-efficacy, diabetes attitudes, and empathy among healthcare providers, but its impact on health professional students is unknown. The purpose of the single-arm pre-post study was to examine the feasibility of this cine-VR diabetes training program as well as to assess changes in cultural self-efficacy, diabetes attitudes, and empathy among health professional students. Participants viewed 12 cine-VR 12 simulations about a 72-year-old patient with type 2 diabetes. Pre-training and post-training, they completed the Transcultural Self-Efficacy Tool, Diabetes Attitude Scale-3, and Jefferson Scale of Empathy. All 92 participants completed the full training. No participants reported technological difficulties or adverse events. For the assessment, 66 participants completed the pre-post measures for a response rate of 71.7% (mean age = 21.1 ± 1.9 years, 82.6% [n = 57] women; 84.1% [n = 58] white). We observed positive improvements in all three cultural self-efficacy subscales: "Cognitive" (t value = -4.705, P < .001), "Practical" (mean change = -.99, t value = -4.240, P < .001), and "Affective" (t value = -2.763, P = .008). Similarly, we observed positive improvements in four of the five diabetes attitude subscales: "Need for special training" (Z = -4.281, P < .001), "Seriousness of type 2 diabetes" (Z = -3.951, P < .001), "Value of tight glucose control" (Z = -1.676, P = .094), "Psychosocial impact of diabetes" (Z = -5.892, P < .001), and "Attitude toward patient autonomy" (Z = -2.889, P = .005). Finally, we observed a positive improvement in empathy (t value = -5.151, P < .001). Findings suggest that the cine-VR diabetes training program has the potential to improve cultural self-efficacy, diabetes attitudes, and empathy among health professional students. A randomized controlled trial is needed to confirm its effectiveness.


"Seeing the Life": Redefining self-worth and family roles among Iraqi refugee families resettled in the United States.
Social and geographic displacement is a global phenomenon that precipitates novel stressors and disruptions that intersect with longstanding familial and social roles. Among the displaced are war-torn Iraqi refugee families, who must address these new obstacles in unconventional ways. This study explores how such disruptions have influenced associations between gender and apparent self-worth experienced by Iraqi refugee families upon relocation to the United States. Further, the psychosocial mechanisms requisite of any novel approach to a new social construct are explored and reveal that production in the family is at the core of instability and shifting power dynamics during resettlement, preventing family members from "seeing the life" in the United States that they had envisioned prior to immigration. Over 200 semi-structured qualitative interviews with Iraqi participants and mental health providers were conducted over the course of the study, and demonstrate a plasticity among social roles in the family and community that transcends the notion of a simple role reversal, and illustrate the complex positionalities that families under stress must approximate during such physical and social displacement.


Metal immobilization and nitrate reduction in a contaminated soil amended with zero-valent iron (Fe0).
Technologies based on zero-valent iron (Fe0) are increasingly being used to immobilize metals in soils and remove metals and nitrate from waters. However, the impact of nitrate reduction on metal immobilization in metal contaminated soils has been poorly investigated so far. Here, different concentrations of Fe0 filings (1%, 2% and 5%; wt%) were applied to a metal contaminated soil. The resulting nitrate reduction and metal (Cd and Zn) immobilization was investigated using a column leaching experiment for 12 weeks. Corrosion of Fe0 filings and precipitation of Fe oxyhydroxydes (FeOOH) on the surfaces of the filings were observed using SEM-EDS and EMPA-WDS at the end of the experiment. Compared to the untreated soil, total nitrate amounts released were lowered by 47%, 59% and 87% in the presence of 1%, 2% and 5% of Fe0, respectively. Concomitantly with nitrate reduction, Cd and Zn concentrations in leachates were strongly alleviated in the presence of Fe0, which was partly attributed to the rise of soil pH subsequent to nitrate reduction. More importantly, biotests with Lupinus albus L. revealed that the mechanisms involved in metal immobilization are stable to root-induced acidification. However, Fe0 was not efficient to reduce Cd concentration in Lolium multiflorum Lam., indicating that root processes other than acidification may re-mobilize metals.


Mesenchymal stem cells in cardiac regeneration: a detailed progress report of the last 6 years (2010-2015).
Mesenchymal stem cells have been used for cardiovascular regenerative therapy for decades. These cells have been established as one of the potential therapeutic agents, following several tests in animal models and clinical trials. In the process, various sources of mesenchymal stem cells have been identified which help in cardiac regeneration by either revitalizing the cardiac stem cells or revascularizing the arteries and veins of the heart. Although mesenchymal cell therapy has achieved considerable admiration, some challenges still remain that need to be overcome in order to establish it as a successful technique. This in-depth review is an attempt to summarize the major sources of mesenchymal stem cells involved in myocardial regeneration, the significant mechanisms involved in the process with a focus on studies (human and animal) conducted in the last 6 years and the challenges that remain to be addressed.


Tissue-specific reductions in mitochondrial efficiency and increased ROS release rates during ageing in zebra finches, Taeniopygia guttata.
Mitochondrial dysfunction and oxidative damage have long been suggested as critically important mechanisms underlying the ageing process in animals. However, conflicting data exist on whether this involves increased production of mitochondrial reactive oxygen species (ROS) during ageing. We employed high-resolution respirometry and fluorometry on flight muscle (pectoralis major) and liver mitochondria to simultaneously examine mitochondrial function and ROS (H2O2) release rates in young (3 months) and old (4 years) zebra finches (Taeniopygia guttata). Respiratory capacities for oxidative phosphorylation did not differ between the two age groups in either tissue. Respiratory control ratios (RCR) of liver mitochondria also did not differ between the age classes. However, RCR in muscle mitochondria was 55% lower in old relative to young birds, suggesting that muscle mitochondria in older individuals are less efficient. Interestingly, this observed reduction in muscle RCR was driven almost entirely by higher mitochondrial LEAK-state respiration. Maximum mitochondrial ROS release rates were found to be greater in both flight muscle (1.3-fold) and the liver (1.9-fold) of old birds. However, while maximum ROS (H2O2) release rates from mitochondria increased with age across both liver and muscle tissues, the liver demonstrated a proportionally greater age-related increase in ROS release than muscle. This difference in age-related increases in ROS release rates between muscle and liver tissues may be due to increased mitochondrial leakiness in the muscle, but not the liver, of older birds. This suggests that age-related changes in cellular function seem to occur in a tissue-specific manner in zebra finches, with flight muscle exhibiting signs of minimising age-related increase in ROS release, potentially to reduce damage to this crucial tissue in older individuals.


IgG and C3 receptors on polymorphonuclear leukocytes in phagocytosis. Role of C3-receptors in the attachment and ingestion phases.
The role of IgG and complement (C3) receptors in the adhesion and ingestion phases of immune complexes by normal human polymorphonuclear leukocytes (PMN) were examined. The immune complexes were sheep red cells (E) sensitized with IgG (EA) or IgM antibodies plus complement (EAC'). Both types of receptor sites are involved in the adhesion phase. Moreover IgG receptors are primarily involved in the ingestion phase. Nevertheless even C3-receptors may be sufficient for complete phagocytosis. Even if EAC' adhesion was still high, C3-receptor specific phagocytosis decreased parallel with the amount of the complement used for EA19S sensibilization. The role of receptor sites on human PMN in adhesion and ingestion phases is discussed.


Expression of genes associated with the antigen presentation and processing pathway are consistently regulated in early Mycobacterium avium subsp. paratuberculosis infection.
The primary objective of this study was to evaluate early gene expression profiles associated with paratuberculosis in cattle exposed to known infectious doses of Mycobacterium avium sub-species paratuberculosis (MAP). A Johne's disease experimental infection field trial was conducted on a mixed population of Holstein and Holstein Red cattle. Blood samples from four MAP exposed and four unexposed cattle, selected based on IFNγ expressions were taken at 9, 13 and 21 weeks and RNA processed to Affymetrix GeneChip™ Bovine Genome arrays. Ontological analysis revealed consistent differences in gene expression between MAP exposed and control animals. A stark variation was observed in expression of a number of genes along antigen presentation pathways, suggesting that MAP exposure potentially results in the host immune response switching to a CD8(+) biased antigen presentation profile. This requires further in-depth analysis since it exposes a hitherto unconfirmed association between MAP exposure and in vivo MHC gene modulation.


A potential nomenclature for the Immuno Polymorphism Database (IPD) of chicken MHC genes: progress and problems.
Among the genes with the highest allelic polymorphism and sequence diversity are those encoding the classical class I and class II molecules of the major histocompatibility complex (MHC). Although many thousands of MHC sequences have been deposited in general sequence databases like GenBank, the availability of curated MHC sequences with agreed nomenclature has been enormously beneficial. Along with the Immuno Polymorphism Database-IMunoGeneTics/human leukocyte antigen (IPD-IMGT/HLA) database, a collection of databases for curated sequences of immune importance has been developed. A recent addition is an IPD-MHC database for chickens. For many years, the nomenclature system for chicken MHC genes has been based on a list of standard, presumed to be stable, haplotypes. However, these standard haplotypes give different names to identical sequences. Moreover, the discovery of new recombinants between haplotypes and a rapid increase in newly discovered alleles leaves the old system untenable. In this review, a new nomenclature is considered, for which alleles of different loci are given names based on the system used for other MHCs, and then haplotypes are named according to the alleles present. The new nomenclature system is trialled, first with standard haplotypes and then with validated sequences from the scientific literature. In the trial, some class II B sequences were found in both class II loci, presumably by gene conversion or inversion, so that identical sequences would receive different names. This situation prompts further suggestions to the new nomenclature system. In summary, there has been progress, but also problems, with the new IPD-MHC system for chickens.


Acute primary purulent pericarditis in an adult patient with unknown X-linked agammaglobulinemia.
X-linked agammaglobulinemia (XLA) is a rare form of inherited immunodeficiency due to an impairment in B-lymphocyte differentiation and maturation. In the majority of cases XLA is diagnosed in childhood, particularly among males affected by recurrent infections and with a family history of immunodeficiency. Infections of respiratory tract, gastrointestinal apparatus, eyes, nose and ears are frequent in XLA patients; on the contrary, infections of myocardium, cardiac valves and pericardium are rarely described in XLA. A 34-year-old man with unknown XLA was hospitalized because of syncope, due to pericardial tamponade, caused by acute primary purulent pericarditis. Immediate pericardiocentesis was effective in improving hemodynamics, and empiric antibiotic therapy was successful in controlling the infection. Purulent pericarditis is a rare disease with high mortality rate: it is usually caused by hematogenous bacterial propagation, direct infection of pericardial space by chest wounds or thoracic surgery, or extension of infection from adjacent tissues. However, this patient had no recent local or systemic infections. Because of unusual clinical picture during hospitalization he underwent further clinical and laboratory evaluations, that showed low immunoglobulin levels. After exclusion of acquired immunodeficiency, genetic tests were performed: they detected deletion of exons 8-9-10 of Bruton Tyrosine Kinase gene on X chromosome, leading to the diagnosis of XLA. Acute purulent primary pericarditis may also occur in adult XLA patients as first clinical manifestation. According to this case report, a primary immunodeficiency syndrome should be considered in patients with atypical cardiac infections and no predisposing conditions, regardless of age.


Early-Onset Colorectal Cancer in Patients under 50 Years of Age: Demographics, Disease Characteristics, and Survival.
Incidence of early-onset colorectal cancer (EO-CRC) is increasing in younger demographics. This study analyzes disease-specific survival in individuals under 50 years of age. Patients with colorectal malignancy were identified in the Surveillance Epidemiology and End Results (SEER) database from 2004 to 2015. Cases were categorized into typically screened (age 50-79 years) and non-typically screened (age 20-49 years) cohorts, as well as by decade. Kaplan-Meier curves and Cox proportional hazard models were used to study survival. A total of 240,772 patients with colorectal cancer were analyzed. Average annual percent change in incidence was -0.24% among typically screened patients and +1.12% among patients with EO-CRC. Patients with EO-CRC more frequently presented with distal tumors (70.6% vs. 57.6%, P < .001) and advanced tumor stage (61.3% vs. 48.6%, P < .001). Patients aged 50 and over had comparable 5 year disease-specific survival to younger patients (68.2% vs. 66.4%, P = .31); however, patients in the 3rd, 4th, and 8th decade of life had particularly low survival rates (59.0% vs. 65.8% vs. 65.8%, logrank P < .001). Patients aged 20-29 years had the most increased risk of cause-specific mortality on univariable Cox regression analysis [HR 1.43, 95% CI 1.31-1.56; P < .001], although this was not significant on multivariable analysis [HR 1.06, 95% CI 0.97-1.15; P = .201]. Male sex, older age, advanced stage, rectal and/or cecal primary, and earlier year of diagnosis were independently associated with increased mortality. Patients with EO-CRC are diagnosed at a later stage and have lower disease-specific survival than those in typically screened cohorts. Additional studies on tumor biology and surveillance strategies are needed to improve outcomes in this population.


Visualizing expression patterns of Shh and Foxf1 genes in the foregut and lung buds by optical projection tomography.
Congenital malformations of the foregut are common in humans. The respiratory and digestive tubes are both derived by division of the foregut primordium. Sonic hedgehog (Shh) and Fork head box F1 (Foxf1) genes encode regulatory molecules that play a pivotal role in gut and lung morphogenesis and are therefore important candidate genes to be examined in models of foregut developmental disruption. Optical projection tomography (OPT) is a new, rapid and non-invasive technique for three-dimensional (3D) imaging of small biological tissue specimens that allows visualization of the tissue distribution of RNA in developing organs while also recording morphology. To explore the application of OPT in this context, we visualized Shh and Foxf1 gene expression patterns in the mouse foregut and lung buds at several stages of development. Time-mated CBA/Ca mice were harvested on embryonic days 9-12. The embryos were stained following whole mount in situ hybridization with labelled RNA probes to detect Shh and Foxf1 transcripts at each stage. The embryos were scanned by OPT to obtain 3D representations of gene expression domains in the context of the changing morphology of the embryo. OPT analysis of Shh and Foxf1 expression in the foregut and lung buds revealed extra details of the patterns not previously reported, particularly in the case of Foxf1 where gene expression was revealed in a changing pattern in the mesenchyme around the developing lung. Shh expression was also revealed in the epithelium of the lung bud itself. Both genes were detected in complementary patterns in the developing bronchi as late as E12, showing successful penetration of molecular probes and imaging at later stages. OPT is a valuable tool for revealing gene expression in an anatomical context even in internal tissues like the foregut and lung buds across stages of development, at least until E12. This provides the possibility of visualizing altered gene expression in an in vivo context in genetic or teratogenic models of congenital malformations.


Kartagener's syndrome seen on CT.
The authors present a case of Kartagener's syndrome that was seen on CT. To the authors' knowledge, this is the first reported case of such. The appearance and interesting aspects of the case will be described.


Diagnostic Imaging of Malignant Tumors in the Orofacial Region.
Though rare, malignancies of the orofacial region often have serious consequences. Malignancies of the orofacial region are typically discovered during a clinical examination or from a patient complaint. Initial discovery from a radiograph is rare. Three-dimensional imaging using advanced imaging techniques often provides adequate information about the aggressive nature of a lesion. Radiographically based imaging demonstrates mainly hard-tissue destruction and, rarely, bone deposition. MRI provides excellent visualization of soft-tissue densities without using ionizing radiation. Functional imaging is used to visualize increased metabolic activity associated with malignancies, and is excellent for determining the metastatic spread of a lesion.


Awareness and readiness to use telemonitoring to support diabetes care among care providers at teaching hospitals in Ethiopia: an institution-based cross-sectional study.
This survey aimed to assess the awareness and readiness of healthcare providers to use telemonitoring (TM) technologies for managing diabetes patients as well as to identify associated factors in Ethiopia. An institution-based cross-sectional quantitative survey was conducted by using a pretested self-administered questionnaire from February to March 2020. Data analysis used a binary logistic regression and partial proportional odds model for factor identification. Randomly selected 423 study physicians and nurses. This study was conducted at the University of Gondar and Tibebe Ghion specialised teaching referral hospitals. Awareness and readiness towards TM in diabetes care. Out of 406 healthcare providers (69.7%, n=283 nurses and 30.3%, n=123 physicians) who completed the survey, 345 (38.7%) heard about TM, when it came to readiness, 321 (25.1%) and 121 (65.5%) of respondents had average and low readiness towards TM, respectively. The result of regression analysis shows that awareness towards TM was higher among respondents who had access to a computer (adjusted OR (AOR): 2.8 (95% CI 1.1 to 7.1)), computer-related training (AOR: 4.6 (95% CI 1.63 to 12.95)) and those who had the experience of supporting patients through digital tools (AOR: 1.7 (95% CI 1.0 to 2.8)). Self-perceived innovators and those who had access to a computer, computer-related training and favourable attitude towards TM had significantly higher readiness to use TM. The findings of this survey revealed low awareness and readiness of participant's towards TM. However, this study suggests the need of improving participant's attitudes, access to smartphones and computers and technical skills to fill this gap.


Chromatin dynamics during seed dormancy.
The chromatin structure determines gene expression and thereby regulates developmental processes in the plant. The molecular mechanisms regulating the induction and release of seed dormancy are still largely unknown and the underlying changes in chromatin organization have hardly been analyzed. Most chromatin studies in plants have been performed on vegetative tissues and have focused on seedlings. The composition of seeds hampers molecular analyses and requires adaptation of the methods that are used for other tissues. Here, we give an overview of the current methods that are used to study different aspects of chromatin organization in seeds. Cytogenetic methods, like fluorescence in situ hybridization and immunolocalization, are used to study chromatin at the microscopic level. Changes in DNA methylation and histone modifications can be studied with molecular methods, like bisulfite sequencing, immunoblotting, and chromatin immunoprecipitation.


Forearm bone mineral density in postmenopausal former users of depot medroxyprogesterone acetate.
The aim of the study was to compare the bone mineral density (BMD) of postmenopausal women who had used depot-medroxyprogesterone acetate (DMPA) or a copper intrauterine device (IUD) as a comparison group until menopause. BMD was measured using dual-energy X-ray absorptiometry at the nondominant forearm for up to 3 years following menopause in 135 women aged 43-58 years: 36 former DMPA users and 99 former IUD users. Mean duration of use was (mean+/-SEM) 9.4+/-3.8 and 14.7+/-6.2 years for the DMPA and IUD groups, respectively. One year after menopause, mean distal radius BMD was 0.435 and 0.449 in DMPA and IUD users, respectively, and 0.426 and 0.447 at 2-3 years following menopause. Ultra-distal BMD was 0.369 and 0.384 in DMPA and IUD users, respectively, at 1 year, and 0.340 and 0.383 at 2-3 years. At 1 and 2-3 years following menopause, no significant differences were observed in the BMD of postmenopausal women aged 43-58 years, who had used DMPA or an IUD until menopause.


Draft Genome Sequence of Anaerolineae Strain TC1, a Novel Isolate from a Methanogenic Wastewater Treatment System.
We report the draft genome sequence of Anaerolineae bacterium strain TC1, newly isolated from a methanogenic wastewater treatment system. The assembly contains 16 contigs in 3 scaffolds representing 3,510,630 bp in total with a G+C content of 41.35%. The genome is predicted to contain 2,793 protein-coding genes and 56 RNAs.


A stop-gain variant in BTNL9 is associated with atherogenic lipid profiles.
Current understanding of lipid genetics has come mainly from studies in European-ancestry populations; limited effort has focused on Polynesian populations, whose unique population history and high prevalence of dyslipidemia may provide insight into the biological foundations of variation in lipid levels. Here, we performed an association study to fine map a suggestive association on 5q35 with high-density lipoprotein cholesterol (HDL-C) seen in Micronesian and Polynesian populations. Fine-mapping analyses in a cohort of 2,851 Samoan adults highlighted an association between a stop-gain variant (rs200884524; c.652C>T, p.R218∗; posterior probability = 0.9987) in BTNL9 and both lower HDL-C and greater triglycerides (TGs). Meta-analysis across this and several other cohorts of Polynesian ancestry from Samoa, American Samoa, and Aotearoa New Zealand confirmed the presence of this association (βHDL-C = -1.60 mg/dL, p HDL-C = 7.63 × 10-10; βTG = 12.00 mg/dL, p TG = 3.82 × 10-7). While this variant appears to be Polynesian specific, there is also evidence of association from other multiancestry analyses in this region. This work provides evidence of a previously unexplored contributor to the genetic architecture of lipid levels and underscores the importance of genetic analyses in understudied populations.


The relationship between physical maltreatment and substance use among adolescents: a survey of 95,788 adolescents in Hong Kong.
To determine the prevalence of corporal punishment and the infliction of injuries from a beating occurring without provocation in the previous 6 months among secondary school children in Hong Kong, and to examine the associations between these two forms of physical maltreatment with substance-use-related behaviors and attitudes. Using secondary data, a cross-sectional, self-administered, anonymous survey of 95,788 secondary school students was conducted in Hong Kong. The prevalence of physical maltreatment showed statistically significant associations with younger age, attendance in Chinese-speaking day schools, temporary housing, residence with only one parent, poorer parental relationship, greater peer influence, perceptions of excessive academic pressure, and feelings of being blamed for poor academic performance. Adolescents who had experienced corporal punishment were more likely to be current users of alcohol (OR = 1.11), tobacco (OR = 1.31), psychoactive substances (OR = 1.60), or heroin (OR = 1.90). Those who had been beaten to injury by a family member without provocation within the past 6 months also were more likely to be current users of alcohol (OR = 1.35), tobacco (OR = 1.65), psychoactive substances (OR = 2.39), and heroin (OR = 3.07). Additionally, students who experienced physical maltreatment were more likely to be acquainted with habitual substance users, have better access to psychoactive substances, to have engaged in sex after abusing drugs, have obtained money from illegal sources to purchase drugs, and believe that psychoactive substances are not harmful or addictive. Physical maltreatment showed strong associations with drug-related behaviors and attitudes, after adjusting for potential confounders. Further longitudinal studies are required to understand the causal direction of the relationship.


Implication of neuropilin 2/semaphorin 3F in retinocollicular map formation.
Neural representations of the environment within the brain take the form of topographic maps whose formation relies on graded expression of axon guidance molecules. Retinocollicular map formation, from retinal ganglion cells (RGCs) to the superior colliculus (SC) in the midbrain, is mainly driven by Eph receptors and their ligands ephrins. However, other guidance molecules participate in the formation of this map. Here we demonstrate that the receptor Neuropilin-2 is expressed in an increasing nasal-temporal gradient in RGCs, whereas one of its ligands, Semaphorin3F, but not other Sema3 molecules, presents a graded low-rostral to high-caudal expression in the SC when mapping is underway. Neuropilin-2 and its coreceptor Plexin A1 are present on RGC growth cones. Collapse assays demonstrate that Semaphorin3F induces significant growth cone collapse of temporal, but not nasal, RGCs expressing high levels of Neuropilin-2. Our results suggest that Neuropilin-2/Semaphorin3F are new candidates involved in retinotopy formation within the SC.


FTO genetic variants and risk of obesity and type 2 diabetes: a meta-analysis of 28,394 Indians.
To investigate the magnitude of association of FTO variants with obesity, type 2 diabetes (T2DM), and related traits among Asian Indians. Random-effect meta-analysis was performed on pooled data from eight studies (n = 28,394) for obesity and related traits and six studies (n = 24,987) for assessment of T2DM risk in Indians where FTO variants were reported. The minor A-allele of the FTO variant rs9939609 was associated with increased risk of obesity (OR 1.15, 95% CI 1.08-1.21, p = 2.14 × 10(-) (5) ), BMI (β = 0.30 kg/m2, 95% CI 0.21-0.38, p = 4.78 × 10(-) (11) ) and other regional adiposity measurements [waist (β = 0.74 cm, 95% CI 0.49-0.99), HC (β = 0.52, 95% CI 0.26-0.78), and waist-hip ratio (WHR) (β = 0.002, 95% CI 0.001-0.004)] in Indians (p ≤ 0.001). An increased risk for T2DM (OR 1.11; 95% CI 1.04-1.19, p = 0.002) was observed, which attenuated when adjusted for age, gender, and BMI (OR 1.09; 95%CI 1.02-1.16, p = 0.01). Our study provides evidence of association between common FTO variant and obesity risk among Indians with comparable effect sizes as in Caucasians. The attenuation of FTO-T2DM risk on BMI adjustment reinforces that BMI does not fully account for the adiposity effects among Asian Indians who are more centrally obese.


Quantification of central metabolic fluxes in the facultative methylotroph methylobacterium extorquens AM1 using 13C-label tracing and mass spectrometry.
The metabolic fluxes of central carbon metabolism were measured in chemostat-grown cultures of Methylobacterium extorquens AM1 with methanol as the sole organic carbon and energy source and growth-limiting substrate. Label tracing experiments were carried out using 70% (13)C-methanol in the feed, and the steady-state mass isotopomer distributions of amino acids derived from total cell protein were measured by gas chromatography coupled to mass spectrometry. Fluxes were calculated from the isotopomer distribution data using an isotopomer balance model and evolutionary error minimization algorithm. The combination of labeled methanol with unlabeled CO(2), which enters central metabolism in two different reactions, provided the discriminatory power necessary to allow quantification of the unknown fluxes within a reasonably small confidence interval. In wild-type M. extorquens AM1, no measurable flux was detected through pyruvate dehydrogenase or malic enzyme, and very little flux through alpha-ketoglutarate dehydrogenase (1.4% of total carbon). In contrast, the alpha-ketoglutarate dehydrogenase flux was 25.5% of total carbon in the regulatory mutant strain phaR, while the pyruvate dehydrogenase and malic enzyme fluxes remained insignificant. The success of this technique with growth on C(1) compounds suggests that it can be applied to help characterize the effects of other regulatory mutations, and serve as a diagnostic tool in the metabolic engineering of methylotrophic bacteria.


Macroscopic on-site evaluation of biopsy specimens for accurate pathological diagnosis during EUS-guided fine needle biopsy using 22-G Franseen needle.
Measuring a visible core length during macroscopic on-site evaluation (MOSE) can be useful for accurate diagnoses during an EUS-guided fine needle biopsy (EUS-FNB). We aimed to estimate visible core cutoff lengths predictive of a correct diagnosis when using 22-gauge Franseen needles for biopsies from pancreatic masses. We assessed 77 consecutive patients who underwent EUS-FNB using 22-gauge Franseen needles for pancreatic masses between March 2018 and October 2018. At least two needle passes were performed in all patients, irrespective of the findings on MOSE. The endoscopists measured the visible cores using a ruler during MOSE. The first two passes were analyzed on a per pass basis, and the correlation between visible core lengths and diagnostic accuracy was evaluated. We evaluated 150 needle passes of 75 patients. The accuracy per pass was 92% (138/150). The median length of the visible cores was 15 (range: 0-60) mm and they were significantly longer in the correct diagnosis group than in the incorrect diagnosis group. The accuracy correlated positively with the visible core length. Receiver-operating characteristic curve analysis of the visible core length for accuracy demonstrated an optimal cutoff value of 10 mm. On multivariate logistic regression, visible core lengths >10 mm independently affected the correct diagnosis (odds ratio: 5.1, P= 0.02). Visible cores exceeding 10 mm may be useful for correct diagnosis while using a 22-gauge Franseen needle for EUS-FNB from pancreatic masses.


Colistin resistance in Escherichia coli confers protection of the cytoplasmic but not outer membrane from the polymyxin antibiotic.
Colistin is a polymyxin antibiotic of last resort for the treatment of infections caused by multi-drug-resistant Gram-negative bacteria. By targeting lipopolysaccharide (LPS), the antibiotic disrupts both the outer and cytoplasmic membranes, leading to bacterial death and lysis. Colistin resistance in Escherichia coli occurs via mutations in the chromosome or the acquisition of mobilized colistin-resistance (mcr) genes. Both these colistin-resistance mechanisms result in chemical modifications to the LPS, with positively charged moieties added at the cytoplasmic membrane before the LPS is transported to the outer membrane. We have previously shown that MCR-1-mediated LPS modification protects the cytoplasmic but not the outer membrane from damage caused by colistin, enabling bacterial survival. However, it remains unclear whether this observation extends to colistin resistance conferred by other mcr genes, or resistance due to chromosomal mutations. Using a panel of clinical E. coli that had acquired mcr -1, -1.5, -2, -3, -3.2 or -5, or had acquired polymyxin resistance independently of mcr genes, we found that almost all isolates were susceptible to colistin-mediated permeabilization of the outer, but not cytoplasmic, membrane. Furthermore, we showed that permeabilization of the outer membrane of colistin-resistant isolates by the polymyxin is in turn sufficient to sensitize bacteria to the antibiotic rifampicin, which normally cannot cross the LPS monolayer. These findings demonstrate that colistin resistance in these E. coli isolates is due to protection of the cytoplasmic but not outer membrane from colistin-mediated damage, regardless of the mechanism of resistance.


Hemolytic anemia accelerated by Babesia spp. infection in splenectomized patient.
A 50-year-old patient presented with severe fatigue, fevers, unexplained weight loss, and night sweats of two-week duration. Within two days, an episode of intravascular hemolysis was noted. Relevant medical history supported a possible diagnosis of Babesia spp. Molecular testing revealed Babesia DNA although the peripheral blood smear did not demonstrate any classic parasite forms. Treatment for Babesia was begun and the patient improved.


Lactate-to-albumin ratio is associated with in-hospital mortality in patients with spontaneous subarachnoid hemorrhage and a nomogram model construction.
Subarachnoid hemorrhage (SAH) is a severe hemorrhagic stroke with high mortality. However, there is a lack of clinical tools for predicting in-hospital mortality in clinical practice. LAR is a novel clinical marker that has demonstrated prognostic significance in a variety of diseases. Critically ill patients diagnosed and SAH with their data in the Medical Information Mart for Intensive Care-IV (MIMIC-IV) database and the eICU Collaborative Research Database (eICU-CRD) were included in our study. Multivariate logistic regression was utilized to establish the nomogram. A total of 244 patients with spontaneous SAH in the MIMIC-IV database were eligible for the study as a training set, and 83 patients in eICU-CRD were included for external validation. Data on clinical characteristics, laboratory parameters and outcomes were collected. Univariate and multivariate logistic regression analysis identified age (OR: 1.042, P-value: 0.003), LAR (OR: 2.592, P-value: 0.011), anion gap (OR: 1.134, P-value: 0.036) and APSIII (OR: 1.028, P-value: < 0.001) as independent predictors of in-hospital mortality and we developed a nomogram model based on these factors. The nomogram model incorporated with LAR, APSIII, age and anion gap demonstrated great discrimination and clinical utility both in the training set (accuracy: 77.5%, AUC: 0.811) and validation set (accuracy: 75.9%, AUC: 0.822). LAR is closely associated with increased in-hospital mortality of patients with spontaneous SAH, which could serve as a novel clinical marker. The nomogram model combined with LAR, APSIII, age, and anion gap presents good predictive performance and clinical practicability.


Neuroprotective efficacy of selective brain hypothermia induced by a novel external cooling device on permanent cerebral ischemia in rats.
This study was aimed at examining whether hypothermia is neuroprotective against permanent cerebral ischemia in rats. A total of 32 male Sprague--Dawley rats were subjected to a middle cerebral artery occlusion. In the hypothermic group, rats (n=10) underwent selective brain hypothermia for 5 hours with the use of a novel surface coil with coolant circulating inside. In the control (n=13) and sham groups (n=9), the rats were maintained at normothermia. After a period of 168 hours ischemia, animals were killed to measure the infarction volume of the brain stained with hematoxylin-eosin. There were no significant differences in physiological parameters except for the temperature. The present style of hypothermia significantly reduced infarction volume in the cortex and caudoputamen. The present results endorse the neuroprotective effect of our method of hypothermia in permanent focal cerebral ischemia at an endpoint of 1 week under the following two conditions: (1) reduction of muscle and caudoputamen temperature to 29 and 31 degrees C, respectively; (2) maintenance of the mean arterial blood pressure above 90 mmHg during hypothermia.


Optimum Thickness Conditions of TiO2 Nanotubes Layer for Efficient Electrochemical Luminescence Cells Application.
We report a TiO2 nanotubes (NTs)-based Electrochemical luminescence (ECL) cell. The ECL cell was fabricated using the electrode of TiO2 NTs and Ru(II) complex (Ru(bpy)2+(3)) as a luminescence materials. The fabricated ECL cell is composed of F-doped SnO2 (FTO) glass/Ru(II)/TiO2 NTs/Ti plate. At a bias voltage of 3 V, the measured ECL efficiencies were 0 Im/W for cell without NTs, 0.03 Im/W for NTs-6.5 µm, 0.07 Im/W for NTs-8 µm and 0.1 Im/W for NTs-10 µm, respectively. The use of Ti02 NTs increases ECL intensities by about 2 times compared to the typical ECL cell without the use of TiO2 NTs.