Concerted interconversion between ionic lock substates of the beta(2) adrenergic receptor revealed by microsecond timescale molecular dynamics.
The recently solved crystallographic structures for the A(2A) adenosine receptor and the beta(1) and beta(2) adrenergic receptors have shown important differences between members of the class-A G-protein-coupled receptors and their archetypal model, rhodopsin, such as the apparent breaking of the ionic lock that stabilizes the inactive structure. Here, we characterize a 1.02 mus all-atom simulation of an apo-beta(2) adrenergic receptor that is missing the third intracellular loop to better understand the inactive structure. Although we find that the structure is remarkably rigid, there is a rapid influx of water into the core of the protein, as well as a slight expansion of the molecule relative to the crystal structure. In contrast to the x-ray crystal structures, the ionic lock rapidly reforms, although we see an activation-precursor-like event wherein the ionic lock opens for approximately 200 ns, accompanied by movements in the transmembrane helices associated with activation. When the lock reforms, we see the structure return to its inactive conformation. We also find that the ionic lock exists in three states: closed (or locked), semi-open with a bridging water molecule, and open. The interconversion of these states involves the concerted motion of the entire protein. We characterize these states and the concerted motion underlying their interconversion. These findings may help elucidate the connection between key local events and the associated global structural changes during activation.


The size-reduced Eudragit® RS microparticles prepared by solvent evaporation method - monitoring the effect of selected variables on tested parameters.
Size-reduced microparticles were successfully obtained by solvent evaporation method. Different parameters were applied in each sample and their influence on microparticles was evaluated. As a model drug the insoluble ibuprofen was selected for the encapsulation process with Eudragit® RS. The obtained microparticles were inspected by optical microscopy and scanning electron microscopy. The effect of aqueous phase volume (600, 400, 200 ml) and the concentration of polyvinyl alcohol (PVA; 1.0% and 0.1%) were studied. It was evaluated how those variations and also size can affect microparticle characteristics such as encapsulation efficiency, drug loading, burst effect and microparticle morphology. It was observed that the sample prepared with 600 ml aqueous phase and 1% concentration of polyvinyl alcohol gave the most favorable results.Key words: microparticles solvent evaporation sustained drug release Eudragit RS®.


Complete L-alanine scan of neuropeptide Y reveals ligands binding to Y1 and Y2 receptors with distinguished conformations.
The synthesis of more than fifty 36-residue oligopeptide analogs of neuropeptide Y (NPY) and their affinity to human Y1 and Y2 receptors is described. Each amino acid of the natural sequence was replaced by L-alanine, the four alanine residues at position 12, 14, 18 and 23 were replaced by glycine. Additional residues were exchanged to closely related ones in order to characterize the prerequisites for binding. A combination of automated single and multiple peptide synthesis using fluoren-9-ylmethoxycarbonyl/tert-butoxy strategy was applied. The purified peptides were characterized by electrospray mass spectrometry, analytical HPLC and amino acid analysis. Binding was investigated by displacement of 125I-labelled neuropeptide Y from human neuroblastoma cell lines SK-N-MC and SMS-KAN. Whereas Pro2 and the integrity of the neuropeptide Y loop is important for the binding to the Y1 receptor, exchanges within the C-terminal helix affect the affinity to the Y2 receptor. The C-terminal pentapeptide amide is important for both receptors and probably represents the binding site. However, Arg33 and Arg35 may not be exchanged by L-alanine in the Y1 system, whereas Arg35 and Tyr36 are the most susceptible residues in the Y2 system. In order to distinguish between conformational effects and direct hormone/receptor interaction via the side chains of neuropeptide Y, circular dichroic studies of the alanine-containing peptides were performed and structure affinity relationships are discussed. Comparing the affinities of the neuropeptide Y analogs to Y1 and Y2 receptors significant differences were found for the two binding sites, which suggests a different active conformation of neuropeptide Y at the two subtypes of receptors. Using molecular dynamics calculations, two distinct conformations were identified which are in good agreement with the data obtained by structure/affinity investigations.


Risks posed by SARS-CoV-2 to North American bats during winter fieldwork.
The virus that causes COVID-19 likely evolved in a mammalian host, possibly Old-World bats, before adapting to humans, raising the question of whether reverse zoonotic transmission to bats is possible. Wildlife management agencies in North America are concerned that the activities they authorize could lead to transmission of SARS-CoV-2 to bats from humans. A rapid risk assessment conducted in April 2020 suggested that there was a small but significant possibility that SARS-CoV-2 could be transmitted from humans to bats during summer fieldwork, absent precautions. Subsequent challenge studies in a laboratory setting have shed new information on these risks, as has more detailed information on human epidemiology and transmission. This inquiry focuses on the risk to bats from winter fieldwork, specifically surveys of winter roosts and handling of bats to test for white-nose syndrome or other research needs. We use an aerosol transmission model, with parameter estimates both from the literature and from formal expert judgment, to estimate the risk to three species of North American bats, as a function of several factors. We find that risks of transmission are lower than in the previous assessment and are notably affected by chamber volume and local prevalence of COVID-19. Use of facemasks with high filtration efficiency or a negative COVID-19 test before field surveys can reduce zoonotic risk by 65 to 88%.


Mitochondrial activity in gametes and transmission of viable mtDNA.
The retention of a genome in mitochondria (mtDNA) has several consequences, among which the problem of ensuring a faithful transmission of its genetic information through generations despite the accumulation of oxidative damage by reactive oxygen species (ROS) predicted by the free radical theory of ageing. A division of labour between male and female germ line mitochondria was proposed: since mtDNA is maternally inherited, female gametes would prevent damages by repressing oxidative phosphorylation, thus being quiescent genetic templates. We assessed mitochondrial activity in gametes of an unusual biological system (doubly uniparental inheritance of mitochondria, DUI), in which also sperm mtDNA is transmitted to the progeny, thus having to overcome the problem of maintaining genetic information viability while producing ATP for swimming. Ultrastructural analysis shows no difference in the conformation of mitochondrial cristae in male and female mature gametes, while mitochondria in immature oocytes exhibit a simpler internal structure. Our data on transcriptional activity in germ line mitochondria show variability between sexes and different developmental stages, but we do not find evidence for transcriptional quiescence of mitochondria. Our observations on mitochondrial membrane potential are consistent with mitochondria being active in both male and female gametes. Our findings and the literature we discussed may be consistent with the hypothesis that template mitochondria are not functionally silenced, on the contrary their activity might be fundamental for the inheritance mechanism. We think that during gametogenesis, fertilization and embryo development, mitochondria undergo selection for different traits (e.g. replication, membrane potential), increasing the probability of the transmission of functional organelles. In these phases of life cycle, the great reduction in mtDNA copy number per organelle/cell and the stochastic segregation of mtDNA variants would greatly improve the efficiency of selection. When a higher mtDNA copy number per organelle/cell is present, selection on mtDNA deleterious mutants is less effective, due to the buffering effect of wild-type variants. In our opinion, a combination of drift and selection on germ line mtDNA population, might be responsible for the maintenance of viable mitochondrial genetic information through generations, and a mitochondrial activity would be necessary for the selective process.


Rose hip and its constituent galactolipids confer cartilage protection by modulating cytokine, and chemokine expression.
Clinical studies have shown that rose hip powder (RHP) alleviates osteoarthritis (OA). This might be due to anti-inflammatory and cartilage-protective properties of the complete RHP or specific constituents of RHP. Cellular systems (macrophages, peripheral blood leukocytes and chondrocytes), which respond to inflammatory and OA-inducing stimuli, are used as in vitro surrogates to evaluate the possible pain-relief and disease-modifying effects of RHP. (1) Inflammatory processes were induced in RAW264.7 cells or human peripheral blood leukocytes (PBL) with LPS. Inflammatory mediators (nitric oxide (NO), prostaglandin E(2) (PGE(2)) and cytokines/chemokines) were determined by the Griess reaction, EIA and multiplex ELISA, respectively. Gene expression was quantified by RT-PCR. RHP or its constituent galactolipid, GLGPG (galactolipid (2S)-1, 2-di-O-[(9Z, 12Z, 15Z)-octadeca-9, 12, 15-trienoyl]-3-O-β-D-galactopyranosyl glycerol), were added at various concentrations and the effects on biochemical and molecular parameters were evaluated. (2) SW1353 chondrosarcoma cells and primary human knee articular chondrocytes (NHAC-kn) were treated with interleukin (IL)-1β to induce in vitro processes similar to those occurring during in vivo degradation of cartilage. Biomarkers related to OA (NO, PGE(2), cytokines, chemokines, metalloproteinases) were measured by multiplex ELISA and gene expression analysis in chondrocytes. We investigated the modulation of these events by RHP and GLGPG. In macrophages and PBL, RHP and GLGPG inhibited NO and PGE(2) production and reduced the secretion of cytokines (TNF-α, IFN-γ, IL-1β, IL-6, IL-12) and chemokines (CCL5/RANTES, CXCL10/IP-10). In SW1353 cells and primary chondrocytes, RHP and GLGPG diminished catabolic gene expression and inflammatory protein secretion as shown by lower mRNA levels of matrix metalloproteinases (MMP-1, MMP-3, MMP-13), aggrecanase (ADAMTS-4), macrophage inflammatory protein (MIP-2, MIP-3α), CCL5/RANTES, CXCL10/IP-10, IL-8, IL-1α and IL-6. The effects of GLGPG were weaker than those of RHP, which presumably contains other chondro-protective substances besides GLGPG. RHP and GLGPG attenuate inflammatory responses in different cellular systems (macrophages, PBLs and chondrocytes). The effects on cytokine production and MMP expression indicate that RHP and its constituent GLGPG down-regulate catabolic processes associated with osteoarthritis (OA) or rheumatoid arthritis (RA). These data provide a molecular and biochemical basis for cartilage protection provided by RHP.


The differential modulation of secondary metabolism induced by a protein hydrolysate and a seaweed extract in tomato plants under salinity.
Climate change and abiotic stress challenges in crops are threatening world food production. Among others, salinity affects the agricultural sector by significantly impacting yield losses. Plant biostimulants have received increasing attention in the agricultural industry due to their ability to improve health and resilience in crops. The main driving force of these products lies in their ability to modulate plant metabolic processes involved in the stress response. This study's purpose was to investigate the effect of two biostimulant products, including a protein hydrolysate (Clever HX®) and a seaweed extract with high amino acids content (Ascovip®), and their combination, on the metabolomics profile of tomato crops grown under salt stress (150 mM NaCl). Several stress indicators (leaf relative water content, membrane stability index, and photosynthesis activity) and leaf mineral composition after salinity stress exposure were assessed to evaluate stress mitigation, together with growth parameters (shoot and root biomasses). After that, an untargeted metabolomics approach was used to investigate the mechanism of action of the biostimulants and their link with the increased resilience to stress. The application of the biostimulants used reduced the detrimental effect of salinity. In saline conditions, protein hydrolysate improved shoot dry weight while seaweed extracts improved root dry weight. Regarding stress indicators, the application of the protein hydrolysate was found to alleviate the membrane damage caused by salinity stress compared to untreated plants. Surprisingly, photosynthetic activity significantly improved after treatment with seaweed extracts, suggesting a close correlation between root development, root water assimilation capacity and photosynthetic activity. Considering the metabolic reprogramming after plant biostimulants application, protein hydrolysates and their combination with seaweed extracts reported a distinctive metabolic profile modulation, mainly in secondary metabolite, lipids and fatty acids, and phytohormones biosynthetic pathways. However, treatment with seaweed extract reported a similar metabolic reprogramming trend compared to salinity stress. Our findings indicate a different mechanism of action modulated by protein hydrolysate and seaweed extract, suggesting stronger activity as a stress mitigator of protein hydrolysate in tomato crops under salinity stress.


Hereditary angioneurotic edema of the larynx.
Hereditary angioneurotic edema (HAE) is an autosomal dominant disease resulting from a deficiency of functional C1-esterase inhibitor. If not recognized promptly and treated properly the disease can result in a fatal outcome as it causes laryngeal edema, which can lead to a life-threatening acute upper airway obstruction. We present the case of a 37-year-old female with HAE of the larynx, who was diagnosed early and treated properly, together with a review of the literature.


Fullerene-containing pillar[n]arene hybrid composites.
The construction and application of fullerene-containing pillar[n]arene organic-inorganic hybrid composites/systems has been discussed and summarized. Both supramolecular interactions and covalent bonds were involved during the preparation process. By using supramolecular interactions, pillar[n]arenes mainly donate the hydrophobic and electron-rich cavity for complexing fullerene and its derivatives. The "bigger-sized" unmodified pillar[10]arene could directly include C60 in a stoichiometry of 1/1 via host-guest interaction, while the perfunctionalized "smaller-sized" pillar[5]arene with an enlarged channel-like structure could further interact with "size-fixed" fullerene. Additionally, recognition site-containing modified fullerene could integrate with the pillararene cavity for fabricating the functional supramolecular system, which could be further used for applications in photodevices. By forming stable covalent bonds, coupled pillar[n]arene and fullerene hybrid composites could be synthesized via either the modification of pillar[n]arenes with the fullerene subunits, or the utilization of fullerene as the partial structural skeleton for building pillar[n]arenes, in which diverse classic organic synthesis reactions were involved. Interestingly, those fullerene-containing pillar[n]arenes with free cavities could further participate in the formation of hierarchical architectures, such as mechanically interlocked molecules, by cooperating with the "dumbbell" molecule.


Intracranial complications of acute otitis media and Gradenigo's syndrome.
We describe the case of a 12-year-old girl with acute otitis media complicated by acute mastoiditis, epidural empyema, thrombosis of the sigmoid sinus and paralysis of the abducens nerve. The patient underwent a mastoidectomy on the left side combined with drainage of the epidural empyema through an extended burr holl and received intravenous antibiotics for 6 weeks and anticoagulation for 12 weeks. This report discusses the intracranial complications of acute otitis media, which were a common problem before the advent of adequate antibiotic drugs but have become rare since their introduction.


Ulinastatin protects rats from sepsis-induced acute lung injury by suppressing the JAK-STAT3 pathway.
Sepsis is usually accompanied by pulmonary inflammations, leading to acute lung injury. During this process, endogenous factors that play a regulatory role could be exploited to therapeutically alleviate such lethal tissue injury. Here, we showed that ulinastatin (UTI) administration could reduce lung tissue necrosis and swelling during sepsis in rats. UTI treatment also decreased the levels of inflammatory mediators both in the lung and in the serum. Mechanistically, we showed that the phosphorylation levels of JAK2 and STAT3 in the lung of UTI-treated rats were lower than control rats and were correlated with the decreased levels of inflammatory mediators. Taken together, these results demonstrate the protective role of UTI in sepsis-induced acute lung injury.


Mechanism of Clostridium difficile toxin A-induced apoptosis in T84 cells.
This study is an investigation into the mechanism of Clostridium difficile toxin A-induced apoptosis in human intestinal epithelial cells. Toxin A induced apoptosis of T84 cells in a dose- and time-dependent fashion. Toxin A-induced apoptosis was completely inhibited by blocking toxin enzymatic activity on Rho GTPases with uridine 5'-diphosphate-2',3'-dialdehyde by a nonspecific caspase inhibitor and was partially inhibited by caspase-1, -3, -6, -8, and -9 inhibitors. Caspases 3, 6, 8, and 9 and Bid activation were detected. Toxin A also induced changes in mitochondrial membrane potential and cytochrome c release at 18-24 h, a time course similar to caspase-9 activation. In conclusion, toxin A induces apoptosis by a mechanism dependent on inactivation of Rho, activation of caspases 3, 6, 8, and 9 and Bid, and mitochondrial damage followed by cytochrome c release. Toxin A proapoptotic activity may contribute to the mucosal disruption seen in toxin A-induced enteritis.


Adult overwhelming meningococcal purpura. A study of 35 cases, 1977-1989.
The study objective was to describe the clinical, biologic, and hemodynamic features of adult overwhelming meningococcal purpura and to examine the prognostic factors by multivariate analysis at the time of admission to the intensive care unit. Thirty-five patients (greater than or equal to 13 years of age) with meningococcal infection, circulatory shock, and generalized purpuric lesions of abrupt onset were recorded in eight intensive care units from 1977 to 1989. The patients were young (mean age, 26.6 years; range, 13 to 68 years) and had been previously healthy. The female-to-male ratio was 3:1. Mortality was 54.3%, with most deaths occurring within the first 48 hours, usually secondary to irreversible shock with multiple organ failure. Ischemic complications (eight cases), prolonged heart failure (seven cases), and secondary septicemia (five cases) were the chief complications among survivors. Initial hemodynamic study after volume loading showed low stroke volume index (mean +/- SD, 29.4 +/- 13 mL/m2) and tachycardia (mean +/- SD, 138 +/- 16 beats per minute), a profile suggesting a greater myocardial depression than usually observed in gram-negative bacillary septic shock. Univariate prognostic analysis showed that four variables at the time of admission were associated with fatal outcome: a plasma fibrinogen level of 1.5 g/L or less, a factor V concentration of 0.20 or less, a platelet count lower than 80 x 10(9)/L, and a cerebrospinal fluid leukocyte count of 20 x 10(6)/L or less. Stepwise regression analysis showed that low fibrinogen level (less than or equal to 1.5 g/L) was the sole adverse prognostic variable (odds ratio = 2, 95% confidence interval, 1.5 to 2.7). Adult overwhelming meningococcal purpura is still associated with high mortality and morbidity. Low fibrinogen level at time of admission may permit early recognition of the most severely ill patients.


The relationship between regional pain with or without neuropathic symptoms and chronic widespread pain.
This study was performed to test whether the risk of developing chronic widespread pain (CWP) in those with regional pain was augmented in those with symptoms of neuropathic pain (NP). Persons free of CWP completed the Douleur Neuropathique 4 (scores ≥3 indicating NP); demographics; Hospital Anxiety and Depression scale; Pittsburgh Sleep Quality Index; and pain medications. Participants were classified as having no pain, regional pain with no symptoms of NP ((Equation is included in full-text article.)), or regional pain with symptoms of NP (NP). At the 12-month follow-up, participants with CWP were identified. Logistic regression estimated the odds ratio, with 95% confidence intervals, of CWP in the (Equation is included in full-text article.)and NP groups compared with no pain, and NP compared with (Equation is included in full-text article.). Partial population attributable risks estimated the proportion of CWP attributable to baseline (Equation is included in full-text article.)or NP exposure. One thousand one hundred sixty-two participants completed the baseline DN4 and provided pain data at follow-up: 523 (45.0%) had no baseline pain, 562 (48.4%) (Equation is included in full-text article.), and 77 (6.6%) NP. One hundred fifty-three (13.2%) had CWP at 12 months: 19 (3.6%) no pain, 108 (19.2%) (Equation is included in full-text article.), and 26 (33.8%) NP. (Equation is included in full-text article.)(2.9 [1.9-4.3]) and NP (2.1 [1.1-4.0]) predicted CWP after adjusting for demographics, Hospital Anxiety and Depression scale, Pittsburgh Sleep Quality Index, and medications. The partial population attributable risk was 41.3% (25.2-54.0) for (Equation is included in full-text article.)and 6.0% (0.1-11.6) for NP. The NP group were not more likely to develop CWP when compared directly with (Equation is included in full-text article.)(1.5 [0.8-2.8]). Neuropathic pain was relatively rare and predicted a small number of new-onset CWP cases. Using these estimates, treatments targeting NP would at best prevent 6% of CWP cases.


Differences in time-domain and spectral indexes of skin-surface laser-Doppler signals between controls and breast-cancer subjects.
There is an urgent need to improve the early diagnosis of breast cancer. The present study applied spectral and beat-to-beat analyses to laser-Doppler (LDF) data sequences measured on the skin surface on the back of the right hands, with the aim of comparing the different peripheral microcirculatory-blood-flow (MBF) perfusion condition between breast-cancer and control subjects. ECG and LDF signals were obtained simultaneously and noninvasively from 23 breast-cancer patients and 23 age-matched control subjects. Time-domain beat-to-beat indexes and their variability parameters were calculated. Spectral indexes were calculated using the Morlet wavelet transform. The beat-to-beat LDF pulse width and its variability were significantly smaller in cancer patients than in the controls. The energy contributions of endothelial-, neural-, and myogenic-related frequency bands were also significantly smaller in cancer patients. The present study has revealed significant differences in the beat-to-beat and spectral indexes of skin-surface-acquired LDF signals between control subjects and breast-cancer patients. This illustrates that LDF indexes may be useful for monitoring the changes in the MBF perfusion condition induced by breast cancer. Since the breast-cancer patients were at TNM stages 0- 2, the present findings may aid the development of indexes for detecting breast cancer.


Development and validation of a new Italian short measure of disgust propensity: The Disgust Propensity Questionnaire (DPQ).
Although a few measures of disgust propensity are available in Italy, most of them take a long time to administer and/or have not shown replicable and sound psychometric properties. In the current study, the authors developed an Italian nine-item self-report measure of disgust propensity (particularly of pathogen disgust)-the Disgust Propensity Questionnaire (DPQ)-to address the limitations of currently available measures. In Study 1, the DPQ was developed through exploratory and confirmatory factor analyses from an initial pool of 33 items that were administered to 784 nonclinical participants. The DPQ showed evidence of an adequate factorial and construct validity as well as internal consistency and temporal stability. In Study 2, additional evidence of the sound psychometric properties of the DPQ was provided by analyzing an independent sample of 315 nonclinical participants and a sample of 208 patients with obsessive-compulsive disorder. This study also showed that the DPQ can discriminate between obsessive-compulsive disorder patients with and without contamination-related concerns, patients with anxiety disorders, and nonclinical participants. An Italian nine-item self-report disgust propensity measure was developed to address the limitations of currently available tools. The Disgust Propensity Questionnaire (DPQ) was evaluated using two independent studies in nonclinical and clinical samples. The DPQ showed adequate factorial and construct validity, internal consistency, and temporal stability. It could discriminate between patients with obsessive-compulsive disorder with contamination-related concerns and all other groups. It is a very short and psychometrically sound measure to assess disgust propensity in Italian samples.


Blunted increase in plasma adenosine levels following dipyridamole stress in dilated cardiomyopathy patients.
Heart failure is characterized by chronically increased adenosine levels, which are thought to express a protective anti-heart failure activation of the adenosinergic system. The aim of the study was to assess whether the activation of adenosinergic system in idiopathic dilated cardiomyopathy (IDC) can be mirrored by a blunted increase in plasma adenosine concentration following dipyridamole stress, which accumulates endogenous adenosine. Two groups were studied: IDC patients (n = 19, seven women, mean age 60 +/- 12 years) with angiographically confirmed normal coronary arteries and left ventricular ejection fraction <35%; and normal controls (n = 15, six women, mean age 68 +/- 5 years). Plasma adenosine was assessed by high-performance liquid chromatography methods in blood samples from peripheral vein at baseline and 12 min after dipyridamole infusion (0.84 mg kg-1 in 10 min). At baseline, IDC patients showed higher plasma adenosine levels than controls (276 +/- 27 nM L-1 vs. 208 +/- 48 nM L-1, P < 0.001). Following dipyridamole, IDC patients showed lower plasma adenosine levels than controls (322 +/- 56 nM L-1 vs. 732 +/- 250 nM L-1, P < 0.001). The dipyridamole-induced percentage increase in plasma adenosine over baseline was 17% in IDC and 251% in controls (P < 0.001). By individual patient analysis, 18 IDC patients exceeded (over the upper limit) the 95% confidence limits for normal plasma adenosine levels at baseline, and all 19 exceeded (below the lower limit) the 95% confidence limits for postdipyridamole plasma adenosine levels found in normal subjects. Patients with IDC have abnormally high baseline adenosine levels and--even more strikingly--blunted plasma adenosine increase following dipyridamole infusion. This is consistent with a chronic activation of the adenosinergic system present in IDC, which can be measured noninvasively in the clinical theatre.


Antenatal magnesium sulfate to prevent cerebral palsy.
Magnesium sulfate given to women before birth at <30 weeks' gestation reduces the risk of cerebral palsy in their children. Our study aimed to assess the impact of a local quality improvement programme, primarily using plan-do-study-act cycles, to increase the use of antenatal magnesium sulfate. After implementing our quality improvement programme, an average of 86% of babies delivered at <30 weeks' gestation were exposed to antenatal magnesium sulfate compared with a historical baseline rate of 63%. Our study strengthens the case for embedding quality improvement programmes in maternal perinatal care to reduce the impact of cerebral palsy on families and society.


Impact of testosterone on cardiac L-type calcium channels and Ca2+ sparks: acute actions antagonize chronic effects.
While androgens generally have been associated with an increased cardiovascular risk, recent studies indicate potential beneficial acute effects of testosterone. However, detailed evaluation of chronic and acute actions of testosterone on the function of cardiac I(Ca,L) and intracellular Ca2+ handling is limited. To clarify this situation we performed whole-cell and single-channel analysis of I(Ca,L), recordings of Ca2+ sparks, measurements of contractility and quantitative real-time RT-PCR in rat cardiomyocytes following testosterone pretreatment and acute testosterone application. Pretreatment with testosterone 100 nM for 24-30 h increased whole-cell I(Ca,L) from 3.8+/-0.8 pA/pF (n=10) to 10.1+/-0.31 pA/pF (n=9) at +10 mV (p<0.001). Increase of I(Ca,L) density was caused by both, increased expression levels of the alpha 1C subunit of L-type calcium channel and a pronounced increment of the single-channel activity (availability 81.8+/-3.15% versus 37.1+/-7.01%; open probability 12.8+/-3.09% versus 1.0+/-0.62%, p<0.01). Moreover, testosterone pretreatment significantly increased the frequency of Ca2+ sparks and improved myocytes contractility without altering SR Ca2+ load. All chronic effects could be inhibited by flutamide. In contrast acute testosterone administration significantly reduced I(Ca,L) density. Indeed, on the single-channel level acute testosterone application completely reversed the chronic testosterone-mediated effects, and antagonized the chronic testosterone effects on Ca2+ spark frequency, which was unaffected by flutamide. Thus, testosterone pretreatment activates I(Ca,L) via nuclear receptor-mediated pathways, while testosterone acutely blocks I(Ca,L) in a direct manner. Thus, testosterone chronically affects the basal level of intracellular Ca2+ handling, which in addition rapidly may be modulated by acute changes of hormone levels.


Organophosphate Pesticide Urinary Metabolites Among Latino Immigrants: North Carolina Farmworkers and Non-farmworkers Compared.
This analysis documents detections and concentrations of the six dialkylphosphate (DAP) urinary metabolite of organophosphorus (OP) pesticides among North Carolina Latino migrant farmworkers, with comparison to non-farmworker Latino immigrants. Participants provided up to four urine samples during the 2012 and 2013 agricultural seasons. Composite urine samples for each year were analyzed. DAP urinary metabolite detections were similar in farmworkers and non-farmworker; for example, for 2012, 75.4% of farmworkers and 67.4% of non-farmworkers and, for 2013, 89.3% of farmworkers and 89.7% of non-farmworkers had dimethylthiophosphate detections. DAP geometric mean concentrations were high; for example, dimethylphosphate concentrations among farmworkers were 11.39 μg/g creatinine for 2012 and 4.49 μg/g creatinine for 2013, while they were 10.49 μg/g creatinine for 2012 and 1.97 μg/g creatinine for 2013 for non-farmworkers CONCLUSIONS:: Research to reduce pesticide exposure among Latino farmworkers and non-farmworkers is needed.


Motivated Use of Numerical Anchors for Judgments Relevant to the Self.
The anchoring effect has been replicated so extensively that it is generally thought to be ubiquitous. However, anchoring has primarily been tested in domains in which people are motivated to reach accurate conclusions rather than biased conclusions. Is the anchoring effect robust even when the anchors are threatening? In three studies, participants made a series of probability judgments about their own futures paired with either optimistic anchors (e.g., "Do you think that the chances that your current relationship will last a lifetime are more or less than 95%?"), pessimistic anchors (e.g., "more or less than 10%?"), or no anchors. A fourth study experimentally manipulated motivation to ignore the anchor with financial incentives. Across studies, anchors that implied high probabilities of unwanted events occurring were ineffective. Together, these studies suggest that anchoring has an important boundary condition: Personally threatening anchors are ignored as a result of motivated reasoning processes.


Distribution of nitric oxide synthases and nitrotyrosine in the kidney of spontaneously hypertensive rats.
To study the cellular distribution and the expression of the major isoforms of NO synthase (NOS) and of nitrotyrosine in the kidney in spontaneous hypertension. We have studied by immunohistochemistry the location of the endothelial (eNOS), neuronal (nNOS) and inducible (iNOS) isoforms and nitrotyrosine in kidney slices from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) using specific antibodies. In order to quantify the expression of these proteins, we have analyzed dissected renal cortical and medullary sections by means of Western blot. Tubular cells were immunoreactive to nNOS and more numerous in the renal medulla of the SHR compared with that of the WKY, specifically in the outer medulla and the papillary region. Western blot also showed higher expression of nNOS in the renal medulla, but not the renal cortex of the SHR. In contrast, iNOS and eNOS distribution and expression were similar in the kidneys of WKY rats and SHR. Immunohistochemistry showed immunoreactive cells to nitrotyrosine in a variety of renal cells similarly distributed in SHR and WKY kidneys. Western analysis detected three proteins of 14.5, 23.7 and 39 kDa immunoreactive to nitrotyrosine, showing a higher expression in the renal cortex compared to the renal medulla. The expression of nNOS is higher in the renal medulla of the SHR, and the distribution of eNOS, iNOS and nitrotyrosine is similar in SHR and WKY rats. It is proposed that the higher expression of the neuronal isoform in the medullary tubular cells is a protective mechanism aimed to improve renal function in spontaneous hypertension.


[A new thermodilution catheter for the determination of heart-minute volume in the arterial system].
A double-lumen polyethylene pigtail catheter (F8, length 100 cm) for the determination of cardiac output (CO) in the arterial system is described. Following femoral insertion of the catheter, 10 ml of ice-cold saline is injected into the left ventricle through the distally ending lumen (0.8 mm2). Through the second lumen (0.8 mm2) which opens 50 cm behind the catheter tip, a thermistor probe (F3, teflon) is advanced into the descending aorta for recording of thermodilution curves. In 21 patients CO was determined first by the Fick method and then by thermodilution in the arterial system. CO by thermodilution correlated favorably with CO by Fick (r = 0.92, SEE 518 ml = 11% of the mean value). There was no systematic deviation from the reference CO. A similarly good correlation existed between stroke volume (SV) by thermodilution and SV by Fick (r = 0.86 SEE 11 ml = 15% of the mean value). Heart rate during thermodilution was 71 bpm, during Fick output estimation 72 bpm (not significant). Following withdrawal of the thermistor probe, simultaneous measurements of left ventricular and aortic pressures can be performed.


Expression and localization of thioredoxin during early implantation in the marmoset monkey.
Thioredoxin is a powerful redox protein expressed in invasive cytotrophoblasts and essential for blastocyst implantation in mice. Isolated marmoset thioredoxin cDNA showed that the deduced amino acid sequence differed from the human sequence by four amino acids. The close homology of thioredoxin in the two species enabled us to use monoclonal antibodies against human thioredoxin to detect marmoset thioredoxin in implantation sites, blastocysts and culture medium. Immunocytochemistry on marmoset implantation sites, on pregnancy days 12 and 15, showed that thioredoxin is highly expressed in uterine luminal epithelium, glands and in some endometrial stromal cells. In attached blastocysts, thioredoxin staining was detected in mural and polar trophoblast cells and both visceral and parietal endoderm, whereas no staining was present in the inner cell mass. A similar pattern of thioredoxin expression was detected in hatched blastocysts attached to Matrigel in tissue culture. Trophoblastic vesicles derived from blastocysts expressed thioredoxin in inner endoderm-like cells and outer trophoblast-like cells and secreted thioredoxin into the culture medium. These experiments have demonstrated thioredoxin expression during early stages of embryo-maternal interaction. We propose that thioredoxin protects the early placenta from oxidative damage and that the marmoset is a valuable model for studying thioredoxin regulation and function during implantation and blastocyst differentiation.


The ellipse sign--an aid in the diagnosis of acute ulcers.
When a persistent collection of barium is seen during a radiographic examination of the upper gastrointestinal tract, differentiation of acute ulceration from non-ulcerating entities (e.g. diverticulum, pseudodiverticulum, residual deformity of surgical intervention) is sometimes difficult. When, however, the persistent collection of barium has an ellipitical configuration, the orientation of the long axis of the ellipse can be an indicator of the nature of the pathologic process. If the long axis is perpendicular to the lumen, the collection represents a deformity without acute ulceration. If the long axis is parallel to the lumen, the collection represents an acute ulceration.


Responses of the central metabolism in Escherichia coli to phosphoglucose isomerase and glucose-6-phosphate dehydrogenase knockouts.
The responses of Escherichia coli central carbon metabolism to knockout mutations in phosphoglucose isomerase and glucose-6-phosphate (G6P) dehydrogenase genes were investigated by using glucose- and ammonia-limited chemostats. The metabolic network structures and intracellular carbon fluxes in the wild type and in the knockout mutants were characterized by using the complementary methods of flux ratio analysis and metabolic flux analysis based on [U-(13)C]glucose labeling and two-dimensional nuclear magnetic resonance (NMR) spectroscopy of cellular amino acids, glycerol, and glucose. Disruption of phosphoglucose isomerase resulted in use of the pentose phosphate pathway as the primary route of glucose catabolism, while flux rerouting via the Embden-Meyerhof-Parnas pathway and the nonoxidative branch of the pentose phosphate pathway compensated for the G6P dehydrogenase deficiency. Furthermore, additional, unexpected flux responses to the knockout mutations were observed. Most prominently, the glyoxylate shunt was found to be active in phosphoglucose isomerase-deficient E. coli. The Entner-Doudoroff pathway also contributed to a minor fraction of the glucose catabolism in this mutant strain. Moreover, although knockout of G6P dehydrogenase had no significant influence on the central metabolism under glucose-limited conditions, this mutation resulted in extensive overflow metabolism and extremely low tricarboxylic acid cycle fluxes under ammonia limitation conditions.


Highly geographical specificity of metabolomic traits among Korean domestic soybeans (Glycine max).
Classification and characterization of agricultural products at molecular levels are important but often impractical with genotyping, particularly for soybeans that have numerous types of variety and landraces. Alternatively, metabolic signature, a determinant for nutritional value, can be the good molecular indicator, which reflects cultivation region-dependent factors such as climate and soil. Accordingly, we analyzed the integrative metabolic profiles of Korean soybeans cultivated in 7 different provinces (representative production areas), and explored the potential association with geographic traits. A total of 210 primary and secondary metabolites were profiled using gas-chromatography time-of-flight mass spectrometry (GC-TOF MS) and liquid-chromatography Orbitrap mass spectrometry (LC-Orbitrap MS). Despite the partial heterogeneity of the soybean varieties, the metabolomic phenotypic analysis based on multivariate statistics inferred the chemical compositional characteristics was primarily governed by the regional specificity. The OPLS-DA model proposed biomarker cluster re-composed with 5 metabolites (tryptophan, malonylgenistin, malonyldaidzin, N-acetylornithine, and allysine) (AUCs = 0.870-1.0). The most distinctive metabolic profiles were identified with the soybeans of Gunsan (middle-western coast) and Daegu (east-southern inland area), which were best characterized by the highest contents of isoflavones and amino acids, respectively. Further interrogation on geographic data suggested the combinatorial association of region-specific metabolic features with general soil texture and climate traits (total rainfall and average annual temperature).


Hepatoprotective effects of chestnut (Castanea crenata) inner shell extract against chronic ethanol-induced oxidative stress in C57BL/6 mice.
This study was carried out to evaluate the protective effects of chestnut inner shell extract (CISE) on chronic ethanol-induced oxidative stress in liver. Mice were fed a control liquid diet (Normal-control), liquid diet containing ethanol alone (EtOH+Vehicle), or were administered CISE and ethanol (EtOH+CISE) for 6 weeks. Administration of ethanol induced liver damage with significant increase of plasma GOT, GPT, hepatic triglyceride (TG) and thiobarbituric acid reactive substance (TBARS) levels. By contrast, co-treatment of CISE with ethanol significantly decreased the activities of GOT and GPT in the plasma, and hepatic TG and TBARS levels. Histological observations were consistent with the result obtained from hepatic lipid quantification. Moreover, CISE treatment with ethanol decreased CYP2E1 expression and increased activities of catalase and superoxide dismutase, which were significantly inhibited by treatment with ethanol alone. To determine the active compound of CISE, fractionation of CISE was conducted and scoparone and scopoletin were identified as main compounds. These compounds were also shown to inhibit the ethanol-induced reduction in antioxidant enzyme activity in an in vitro model system. These results suggest that CISE has protective effects against ethanol-induced oxidative damage, possibly by inhibition of lipid accumulation, peroxidation and increase of antioxidant defense system in the liver.


Investigations of milk quality from teats with milk flow disorders.
The objective of this study was to investigate the quality of milk from teats with milk flow disorders. Somatic cell count, pathogens, and signs of mastitis (>100,000 cells/ml and pathogens detected) were determined in the milk from all teats of the udder before treatment of the affected teat, as well as 1 and 6 mo later. Teats with milk flow disorders were compared to all of the other teats from the same udder. Before treatment, the SCC from affected teats was 4.3 million higher, the odds of detecting pathogens 6 times higher, and the odds of mastitis 11 times higher than in control teats (when adjusted for other significant explanatory variables). SCC and the risk of mastitis decreased after surgical treatment of the affected teats, whereas the chance of detecting pathogens was not affected. Six months after treatment, the SCC was 1.3 million higher, and the odds of mastitis 6.5 times higher than in control teats. Throughout the study period neither SCC, the odds of detecting pathogens, nor mastitis changed significantly in control teats. It may be concluded from this study that milk quality from teats with milk flow disorders is decreased before treatment and does not reach the milk quality from unaffected teats within 6 mo after treatment.


[The energy requirements of Holstein-Friesian heifers during rearing and pregnancy under the tropical conditions of Cuba. 1. Determination of dry matter and energy assimilation and of the live weight development and energy expenditure].
In an individual feeding experiment with 48 Holstein-Friesian heifers divided into 3 groups, the effect of various energy levels on live weight development and energy expenditure in the age bracket of 97 days (78 kg live weight) to conception (350 kg LW) and from conception to calving (500 kg LW) was investigated. Rations with differing concentrate--roughage ratios were used. The differentiation of energy intake per day between groups 1 and 2 and group 3 amounted to 7.5% in the 1st development period and to 20% between group 1 and groups 2 and 3 in the 2nd development period. There were significant differences in the live weight gain of 95 g/animal and day in the first development period (up to conception) and of 81 g/animal and day in the second period of development (from conception to calving). Under the tropical climatic conditions the energy expenditure in these studies was approximately 10% under the energy expenditure recommended by the NRC (1978). While there was a clear reciprocal relation between energy expenditure and live weight gain in the first period of development, this relation was less distinct in the second period of development.


Allogeneic adipose-derived stem cells with low immunogenicity constructing tissue-engineered bone for repairing bone defects in pigs.
The ideal cells for tissue engineering should have the following characteristics: easy obtainment, safety, immune privilege, the capability of self-renewal, and multipotency. Adipose-derived stem cells (ADSCs) are a promising candidate. However, the immunogenicity of allogeneic mesenchymal stem cells limits their long-term benefits. In this study, we introduced human cytomegalovirus US2/US3 gene into the ADSCs to decrease the expression of MHC I protein of ADSCs and reduce the activation of T-cells of the recipient animals. Moreover, the biosafety and biological characteristics of ADSCs transfected with the US2/US3 genes (ADSCs-US2/US3) were similar to normal ADSCs. Then we took ADSCs-US2/US3 to construct a tissue-engineered bone for repairing bone defects in pigs and found that there were no great differences in repair effects or healing time between the allogeneic ADSCs-US2/US3 group and the autologous ADSC group. These results suggest that allogeneic ADSCs-US2/US3 have the advantages of biological safety, low immunogenicity, and effective osteogenesis. Such barely immunogenic ADSCs will be crucial for the success of future tissue-regenerative approaches.


Accurate predictions of the electronic excited states of BODIPY based dye sensitizers using spin-component-scaled double-hybrid functionals: a TD-DFT benchmark study.
The vertical excitation energies of 13 BODIPY based dye sensitizers are benchmarked by means of TD-DFT, using 36 functionals from different DFT rungs. Most TD-DFT results were found to overestimate the excitation energies, and show mean absolute error (MAE) values in the range 0.2-0.5 eV. The dispersion-corrected, spin-component-scaled, double-hybrid (DSD) functionals DSD-BLYP and DSD-PBEP86 were found to have the smallest MAE values of 0.083 eV and 0.106 eV, respectively, which is close to the range of average errors found in the more expensive coupled-cluster methods. Moreover, DSD-BLYP and DSD-PBEP86 functionals show excellent consistency and quality of results (standard deviation = 0.048 eV and 0.069 eV respectively). However, the range separated hybrid (RSH) and the range separated double hybrid (RSDH) functionals were found to provide the best predictability (linear determination coefficient R 2 > 0.97 eV).


[Chronic Budd-Chiari syndrome caused by fibrotic stricture of the inferior vena cava].
Author elaborates the case of a male patient, 49, who had chronic Budd-Chiari syndrome due to stricture caused by a valve in the vena cava inferior, and who bled to death as a result of the rupture of the oesophagus varix. In the 4-year-course of his disease serious portal hypertension and extreme degree of ascites were most characteristic. In the light of special literature author treats the etiology of the disease, its clinical characteristics and diagnostic possibilities. Attention is directed to this rare but operable illness, since literature bears witness to the increasing number of successful operations in this field.


Labyrinthitis ossificans.
Three cases with postinflammatory inner ear sequelae are presented to illustrate unusual histopathologic changes. Endolymphatic hydrops without changes in the perilymphatic system was present in one ear following "influenza" meningitis and labyrinthitis ossificans in the contralateral ear. The characteristic histopathological changes of the temporal bones with hematogenic bacterial infection were an extensive labyrinthine ossification associated with a generalized sclerotic change of the whole periotic bone. Bony fixation of the stapedial footplate occurred with the generalized inflammatory process of the otic capsule. Severe and diffuse labyrinthitis ossificans occurred in one case due to tympanogenic inflammation spreading through the round window membrane in the course of suppurative otitis media. A general immunosuppression leading to fatal termination was the apparent factor predisposing to the inner ear complication.


How Should Engineered Nanomaterials Be Regulated for Public and Environmental Health?
A central ethical and policy issue regarding minimizing and managing risks of engineered nanomaterials (ENMs) is whether existing legal frameworks sufficiently protect public health and the environment. This article argues that policymakers should (1) use existing laws to regulate ENMs and the best available evidence to inform appropriate levels of regulation and (2) support additional research on risks of ENMs. Were they to do so, public health and environmental risks of ENMs could be minimized and managed without sacrificing their potential clinical, social, and economic benefits.


Erythema Migrans-like COVID Vaccine Arm: A Literature Review.
COVID Vaccine Arm (CVA) is an adverse drug reaction from mRNA vaccine for SARS-CoV-2. CVA is characterized by erythema and edema on the vaccination site (usually deltoid area) that appears from 5 to 10 days after vaccination and is sometimes associated with itching or pain. The exact etiology of CVA is still unclear, but delayed hypersensitivity against an excipient seems to play an essential role in the pathogenesis of the disease. This work performs a systematic literature review on CVA using three different databases containing articles published until 10 November 2021. The literature review includes eight papers reporting single cases or case series of CVA. Moreover, it also addresses, other cutaneous reactions following COVID 19 vaccinations as well as possible differential diagnosis. CVA migrans-like erythema is characterized by a ring-shaped rash in the injection area, which appears some days after the injection and disappears in about 10 days. This reaction may appear more rapidly in subsequent doses.


Olfactory neuroepithelium in the superior and middle turbinates: which is the optimal biopsy site?
Olfactory neuroepithelium (ON) biopsy has several therapeutic applications for both disorders of olfaction and neurodegenerative diseases. Successful collection of ON is still anything but routine due to a dearth of studies on the distribution of ON in the superior and middle turbinates. To determine the location in which ON is most likely to be present in endoscopically removed cadaver superior and middle turbinates as well as the influences of gender, age, and naris side on the presence of ON and the extent to which it is present. We conducted a prospective anatomical study. The superior and middle turbinates on both sides endoscopically removed from 25 fresh cadavers (less than 12 h post-mortem). The turbinates were halved into anterior and posterior segments for a total of 200 specimens, which were analyzed after hematoxylin and eosin and immunohistochemical staining. Hematoxylin and eosin-stained slides were subjected to blind examination by 3 independent pathologists, and the presence of ON was graded on a 5-point scale from 0 to 4. Kappa measurement was used to determine the agreement between pairs of observers. ON was present in 82.9% of superior turbinate samples and in 17.1% of middle turbinate samples. Immunohistochemistry detected ON in superior turbinates only by S-100 staining and only in 15 fragments. Gender, age, and naris side had no statistically significant effects on the presence of ON. When biopsying ON, the posterior portion of the superior turbinate should be targeted whenever possible because it has the highest concentration of ON among the nasal structures.


Left atrial appendage emptying velocity does not predict postoperative atrial fibrillation in patients undergoing cardiopulmonary bypass.
Atrial fibrillation is a common complication of cardio-pulmonary bypass and improved pre-operative risk assessment could help guide prophylactic therapy. This study examined whether reduced left atrial appendage flow velocities measured by transoesophageal echocardiography pre-operatively in patients in sinus rhythm predicted development of postoperative atrial fibrillation. All patients who underwent transoesophageal echocardiography for clinical indications with measurements of left atrial appendage velocities within twelve months prior to cardio-pulmonary bypass were retrospectively identified. Postoperative records were reviewed and the patients divided into two groups based on the presence or absence of clinically significant atrial fibrillation during hospitalization following cardio-pulmonary bypass. Thirty-six patients (mean age 61.1 +/- 14.8 years, 18M/18F) were included in the study. The overall incidence of atrial fibrillation in the cohort was 17/36 patients (47%). Mean left atrial appendage emptying velocity was 50.8 +/- 23.3 cm/s2 (range 26-119) in the patients with sinus rhythm only and 41.5 +/- 16.7 cm/s2 (range 16-76), in the patients with postoperative atrial fibrillation (P=ns). In our patient population there was no significant difference in left atrial appendage emptying velocity measured by transoesophageal echocardiography in patients with and without postoperative atrial fibrillation. Pre-operative measurement of left atrial appendage emptying velocity cannot be relied upon to risk stratify patients prior to cardio-pulmonary bypass.


Experimental Study of Adsorption Characteristics and Deformation of Coal for Different Gases.
Coal seam deformation due to gas adsorption affects the stability of the underground structure. Natural coal blocks of the Shanxi Formation were selected to study the dynamic adsorption characteristics of coal samples subjected to CO2, CH4, and N2 gas injections under coaxial pressure and confining pressure (7 MPa), as well as the displacement of CH4 with CO2 and N2 under the same conditions. The results show that, under the same conditions, the strain in the coal samples first increased, followed by a rapid increase along with the increase in pressure, with the transverse strain being always higher than the axial strain. The amount of gas adsorption varied from high to low as CO2 > CH4 > N2, and the final adsorption strains and equilibrium times were different for each gas. Based on the increase in gas pressure, the gas adsorption strain curve can be divided into two stages. The displacement of N2 only uses partial pressure to achieve the desorption of CH4 in the coal sample, leading to shrinkage deformation of the coal sample. In contrast, the displacement of CO2 has the dual effects of competitive adsorption and partial pressure reduction on CH4, leading to the swelling deformation of the coal sample.


Recycling of MUC1 is dependent on its palmitoylation.
MUC1 is a mucin-like transmembrane protein expressed on the apical surface of epithelia, where it protects the cell surface. The cytoplasmic domain has numerous sites for phosphorylation and docking of proteins involved in signal transduction. In a previous study, we showed that the cytoplasmic YXXphi motif Y20HPM and the tyrosine-phosphorylated Y60TNP motif are required for MUC1 clathrin-mediated endocytosis through binding AP-2 and Grb2, respectively (Kinlough, C. L., Poland, P. A., Bruns, J. B., Harkleroad, K. L., and Hughey, R. P. (2004) J. Biol. Chem. 279, 53071-53077). Palmitoylation of transmembrane proteins can affect their membrane trafficking, and the MUC1 sequence CQC3RRK at the boundary of the transmembrane and cytoplasmic domains mimics reported site(s) of S-palmitoylation. [3H]Palmitate labeling of Chinese hamster ovary cells expressing MUC1 with mutations in CQC3RRK revealed that MUC1 is dually palmitoylated at the CQC motif independent of RRK. Lack of palmitoylation did not affect the cold detergent solubility profile of a chimera (Tac ectodomain and MUC1 transmembrane and cytoplasmic domains), the rate of chimera delivery to the cell surface, or its half-life. Calculation of rate constants for membrane trafficking of wild-type and mutant Tac-MUC1 indicated that the lack of palmitoylation blocked recycling, but not endocytosis, and caused the chimera to accumulate in a EGFP-Rab11-positive endosomal compartment. Mutations CQC/AQA and Y20N inhibited Tac-MUC1 co-immunoprecipitation with AP-1, although mutant Y20N had reduced rates of both endocytosis and recycling, but a normal subcellular distribution. The double mutant chimera AQA+Y20N had reduced endocytosis and recycling rates and accumulated in EGFP-Rab11-positive endosomes, indicating that palmitoylation is the dominant feature modulating MUC1 recycling from endosomes back to the plasma membrane.


Mobile 240 kVp phototimed chest radiography.
Chest radiographs obtained with mobile equipment for 240 kVp phototimed technique and conventional technique were compared in 161 patients. Features of interest were visualization of air space, interstitium, mediastinum, pleura and vessels, tracheobronchial tree, catheters and tubes. The 240 kVp system demonstrated consistently good visualization of central air passages, hilar contours, and pulmonary detail. It was superior to the conventional portable in eliminating respiratory motion, defining lung infiltrates, and localizing the endotracheal tube relation to the carina. The 240 kVp system had noticeable deficiencies in depicting calcifications, bone detail, and fat planes. These deficiencies were not of clinical significance. Although the equipment was compact, easily maneuverable, and simple to operate, the cassette holder was somewhat heavy--a detrimental but manageable factor. Another shortcoming was poor visualization of homogeneously opacified catheters within the mediastinum, although catheters with a dense radiopaque stripe alleviated this problem.


Stress concentrations and bone microdamage: John Currey's contributions to understanding the initiation and arrest of cracks in bone.
The microarchitecture of bone tissue presents many features that could act as stress concentrators for the initiation of bone microdamage. This was first identified by John Currey in a seminal paper in 1962 in which he presented the mechanical and biological evidence for stress concentrations at the bone surface, within the bone through the action of stiffness differentials between architectural features including between lamellae, and at the level of the lacunar and canalicular walls. Those early observations set the stage to consider how microscopic damage to bone tissue might affect the properties of bone at a time when most in the scientific community dismissed microcracks in bone as artifact. Evidence collected in the nearly 60 years since those important initial observations suggest that some of these architectural features in bone tissue are more effective as crack arrestors than as crack initiators. Sites of higher mineralization in the bone matrix, particularly interstitial sites in both cortical and trabecular bone, may serve preferentially as locations for crack initiation, whereas those boundaries identified by Currey as both stress concentrators and stress arrestors are more effective at stopping cracks than at initiating them.


Effects of tree species and topography on soil and microbial biomass stoichiometry in Funiu Mountain, China.
Soil and microbial biomass stoichiometry plays an important role in understanding nutrient cycling in terrestrial ecosystems. However, studies on soil and microbial biomass stoichiometry in forests are rare. This study investigated the effect of tree species and topographic factors on the ecological stoichiometry of soil and soil microbial biomass. Three types of forest stands (Quercus variabilis, Larix principis-ruprechtii, and Cotinus coggygria Scop.) in the Beiru River basin of Funiu Mountain were analyzed in September 2018. Six slope positions (sunny bottom slope, sunny middle slope, sunny top slope, shady bottom slope, shady middle slope, and shady top slope) were selected, and the total number of sampling plots was 108. The stoichiometric indices of soil and microbial biomass were determined. At a depth of 0-10 cm, the soil organic C contents in different stands followed the order of C. coggygria (27.7 ± 5.2 g/kg) > Q. variabilis (24.5 ± 4.9 g/kg) > L. principis-ruprechtii (20.8 ± 4.3 g/kg) (P < 0.05). The soil organic C contents at depths of 0-10 cm with different slope aspects and at different slope positions also showed significant differences (P < 0.05). The highest MBC content was observed at the slope bottom (1002 ± 157 mg/kg), whereas the lowest was observed at the slope top (641 ± 98.3 mg/kg). Redundancy analysis showed that the contribution of tree species to these differences was 57.1%, whereas that of topographical factors was 36.2%. Tree species more significantly affected soil nutrients and microbial biomass C, N and P than did topographic factors.


Spontaneous interbody fusion after controlled injuries to the spine: an experimental study in rabbits.
To evaluate the rationale of spontaneous spine fusion after a spinal injury, the authors conducted an experimental study that consisted of three types of controlled injuries to a rabbit spine model. The first was injury to the intervertebral disk (type I injury). The second was injury of the intervertebral disk along with injury to one of the adjacent vertebral end plates (type II). In type III injury, both the opposing end plates were injured along with the intervertebral disk. In 38 rabbits, a total of 82 injuries of these three types were inflicted. Twenty-six injuries were of type I (n = 22 rabbits), 26 were type II (n = 24 rabbits), and 30 were type III (n = 26 rabbits). Spontaneous fusion occurred only in type III injuries. From the 30 type III injuries, fusion occurred in 20 (66.6%). For an autofusion to occur, both epiphyseal plates may be injured. In the clinical situation, this observation suggests that a radiographically obscure lesion of both neighboring vertebrae may proceed to autofusion of that spinal segment observed later.


The effect of different postweaning altrenogest treatments of primiparous sows on follicular development, pregnancy rates, and litter sizes.
This study investigated follicular development during and after postweaning altrenogest treatment of primiparous sows in relation to subsequent reproductive performance. Primiparous sows (n = 259) were randomly assigned at weaning (d 0) to 1 of 4 groups: control (no altrenogest, n = 71), RU4 (20 mg of altrenogest from d -1 to 2, n = 62), RU8 (20 mg of altrenogest from d -1 to 6, n = 65), or RU15 (20 mg of altrenogest from d -1 to 13, n = 61). Average follicular size (measured by ultrasound) increased during altrenogest treatment and resulted in larger follicles at the start of the follicular phase for RU4, RU8, and RU15 compared with controls (5.3 ± 0.9, 5.5 ± 1.3, 5.1 ± 1.2, and 3.4 ± 0.6 mm, respectively; P < 0.0001). Farrowing rate was greater in RU15 (95%) than in RU8 (76%; P = 0.04). The RU15 group also had more piglets (2 to 3 more piglets total born and born alive; P < 0.05) than the other treatment groups. Follicular development at weaning clearly affected reproductive performance. At weaning, average follicular size: small (<3.5 mm), medium (3.5 to 4.5 mm), or large (≥ 4.5 mm), was associated with farrowing rates of 86, 78, and 48%, respectively (P < 0.001). Sows with large follicles at weaning had low farrowing rates (71%) in RU4, very low farrowing rates (22%) in RU8, but normal farrowing rates in RU15 (83%). In conclusion, this study showed that 15 d of postweaning altrenogest treatment of primiparous sows may allow follicle turnover in sows that had large follicles at weaning and that this was associated with an improved reproductive performance. It also showed that shorter treatment with altrenogest (4 or 8 d) is beneficial for sows with small follicles at weaning, but is not recommendable for sows with large follicles at weaning.


Pharmaceutical Development of Intraperitoneal Arachis hypogaea as a Renal Protective Agent.
Kidneys are among the vital organs of the human body; therefore, damage from any exogenous/endogenous agent may put human life at risk. Arachis hypogaea (AH) contains different free radical scavenging flavonoids, stilbenes, and tannins. This research aimed to elucidate the possible nephroprotective mechanism of AH methanolic crude extract (AHcr) and n-hexane oil fraction (AHO) against gentamycin-induced nephrotoxicity. After the extraction of the crude oil of the plant, they were tested against a Gentamycin (GM)-treated group of Swiss Albino mice for their nephroprotective action. Animals were divided into six (6) equal groups with five (5) animals in each group. These groups were: control group (0.5 mL normal saline via intraperitoneal -i.p), gentamycin group (gentamycin 100 mg/kg i.p), Silymarin + gentamycin group (Silymarin 50 mg/kg and gentamycin 100 mg/kg i.p), plant extract (AHcr1) and gentamycin group (AHcr1 250 mg/kg and gentamycin 100 mg/kg i.p), AHcr2 + gentamycin group (AHcr2; 500 mg/kg and gentamycin 100 mg/kg i.p) and the hexane oil fraction (AHO) + gentamycin (AHO 1 mL/kg and GM 100 mg/kg i.p). After completion of doses, animals were sacrificed for the collection of blood to further investigate biochemical changes and histopathological changes in kidney tissues. Serum creatinine, urea, and blood urea nitrogen significantly increased (p < 0.001) in the gentamycin-treated group as compared to the control group. The elevated level of serum creatinine, urea, and blood urea nitrogen was decreased significantly (p < 0.001) in groups treated with AHcr and AHO compared to the gentamycin group. Similarly, the histopathological study of kidney tissues from the gentamycin group showed tubular necrosis, vacuolation, and fibrosis. The effect of crude extract and hexane soluble fraction of AH caused a significant reversal of gentamycin-induced nephrotoxicity.


The Daniel K. Inouye College of Pharmacy Scripts: Precision Medicine Through the Use of Pharmacogenomics: Current Status and Barriers to Implementation.
The precision medicine initiative brought forth by President Barack Obama in 2015 is an important step on the journey to truly personalized medicine. A broad knowledge and understanding of the implications of the pharmacogenomic literature will be critical to the achievement of this goal. While a great amount of data has been published in the areas of pharmacogenomics and pharmacogenetics, there are still relatively few instances in which the need for clinical intervention can be stated without doubt, and which are widely accepted and practiced by the medical community. As our knowledge base rapidly expands, issues such as insurance reimbursement for genetic testing and education of the health care workforce will be paramount to achieving the goal of precision medicine for all patients.


The parasitic barnacle Peltogaster reticulata Shiino, 1943 (Rhizocephala,<br />Peltogastridae) from Russian waters of the Sea of Japan: morphological description, molecular identification and complete larval development.
A rhizocephalan parasite of the hermit crab Pagurus minutus Hess, 1865 (Decapoda, Paguridae) is described from Russian waters using morphological and molecular methods. Partial sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI) were used as a genetic marker. We refer this species to Peltogaster reticulata Shiino, 1943 (Rhizocephala, Peltogastridae) based on its morphological description, since genetic data for the species from its type locality are lacking. The species can be distinguished from its known congeners by the reticular pattern of the external cuticle. Peltogaster reticulata is the sister taxon to P. postica Yoshida Osawa, 2011 (in Yoshida et al. 2011), inhabiting the same host in southwestern Japan. The complete larval development of Peltogaster reticulata, including five naupliar and one cypris stage, is described and illustrated using SEM.


The Unethical Use of Paracetamol As a Food Tenderizer in Four Selected African Countries: A Major Public Health Concern?
Paracetamol poisoning is the commonest cause of acute liver injury. Therefore, the unethical use of paracetamol as a food tenderizer poses a threat to human health. Although this is a common practice in Ghana, Uganda, Nigeria, and Kenya, there are few or no scientific records on the use of paracetamol as a food tenderizer and its deleterious effects, thus making it difficult to regulate this practice. This review aims to fully collate and present a systematic overview of the literature on the use of paracetamol as a food tenderizer in these countries, the potentially harmful effects posed by the practice, and measures in place to curb the situation. Additionally, this review aims to reveal the scientific gaps and areas requiring more research, thus providing a reference for further research to regulate this unscrupulous practice. From our extensive review of the literature, the high cost of fuel used in cooking and longer cooking times are the main reasons for the inappropriate use of paracetamol as a food tenderizer. Also, this review concludes that little has been done to create public awareness of this unethical practice. Furthermore, few ways to monitor, control and regulate this practice have been proposed.


Effects of service provider attitudes and employment status on citizenship behaviors and customers' attitudes and loyalty behavior.
The relationship among job satisfaction, affective commitment, service-oriented organizational citizenship behaviors (OCBs), customer satisfaction, and customer loyalty were examined for a sample of 249 hairstylists and 1 of their corresponding customers. Employee satisfaction was positively related to service-oriented OCBs, customer satisfaction, and customer loyalty, whereas affective commitment was not related to these outcomes. The extent to which the predictor variables interacted with one another and the role of employment status on these relationships was also explored. High levels of job satisfaction or affective commitment resulted in more service-oriented OCBs for employees and self-employed workers, whereas high levels of both resulted in more service-oriented OCBs for owners.


PRMT1 negatively regulates activation-induced cell death in macrophages by arginine methylation of GAPDH.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is implicated in cell death in addition to a role as a glycolytic enzyme. In particular, when cells are exposed to cellular stressors involving nitric oxide (NO) production, GAPDH can undergo NO-induced S-nitrosylation and S-nitrosylated GAPDH has been shown to elicit apoptosis. However, the mechanism underlying the regulation of the pro-apoptotic function of GAPDH remains unclear. Here, we found that protein arginine methyltransferase 1 (PRMT1) mediated arginine methylation of GAPDH in primary bone marrow-derived macrophages in a NO-dependent manner. Moreover, PRMT1 inhibited S-nitrosylation of GAPDH as well as its binding to SIAH1, thereby reducing the nuclear translocation of GAPDH in lipopolysaccharide (LPS)/interferon (IFN)-γ-activated macrophages. Furthermore, depletion of PRMT1 expression by RNA interference potentiated LPS/IFN-γ-induced apoptosis in macrophages. Taken together, our results suggest that PRMT1 has a previously unrecognized function to inhibit activation-induced cell death of macrophages through arginine methylation of GAPDH.


Childhood trauma, borderline personality symptomatology, and psychophysiological and pain disorders in adulthood.
In the empirical literature, there is support for the idea of a relationship between childhood trauma and various psychophysiological as well as pain disorders, and between borderline personality symptomatology and somatic preoccupation, as well as chronic pain. However, to date, no single study has examined the relationships between childhood trauma and borderline personality symptomatology, and the number of psychophysiological and pain disorders in adulthood. In this study, the authors examined these relationships and found no significant correlation between borderline personality symptomatology and the number of psychophysiological and pain disorders. However, there were positive and significant correlations between childhood physical abuse, emotional abuse, and witnessing violence and the number of psychophysiological and pain disorders in adulthood. Using hierarchical-regression analyses to determine which specific childhood traumas significantly predicted the number of psychophysiological and pain disorders, only witnessing violence emerged, accounting for 12% of the variance. These data suggest that a general factor associated with various forms of trauma predicts number of psychophysiological and pain disorders and that a specific predictor may be witnessing violence in childhood.


Potentiation of cytotoxic activity of immunotoxins on cultured human cells.
The cytotoxic activity against human tumor cells of toxic conjugates of Pseudomonas exotoxin with anti-transferrin receptor antibody or epidermal growth factor was potentiated up to 10 to 20-fold by the calcium antagonists verapamil, D-600, and diltiazem and by the lysosomotropic agent beta-glycylphenyl-naphthylamide. The potentiating activity of these agents could be predicted by measuring the inhibition of protein synthesis by the immunotoxins on various cell lines. The use of potentiating agents such as these in combination with immunotoxins may prove useful in the treatment of some human cancers.


Bacterial diversity in the human saliva from different ages.
To obtain primary idea on oral bacterium species that are generally present in periodotally healthy Koreans, the oral bacterial flora in the saliva of four periodontally healthy Koreans at different ages (5, 32, 35, 65) was investigated in this study. For this investigation, 16SrRNA gene clone libraries were generated from the saliva of the four healthy Koreans, and 50 clones were randomly selected from each saliva clone library and sequenced. Totally, 37 different kinds of bacterial 16S rRNA gene sequences were identified based on sequence homology search through GenBank database. The 37 kinds of saliva clone sequences were classified to 14 genera and 2 uncultured and 1 unidentified bacteria. Among the 14 identified genera, Streptococcus, Prevotella, and Veillonella were common genera, and Streptococcus was dominant genus that accounted for 7 different species. Among the seven Streptococcus species, S. salivarius appeared as the most common species. More numbers of species belonging to the genera Streptococcus and Prevotella was present in saliva from ages 32 and 35. While saliva from ages 5 and 65 showed more numbers of species belonging to the genera Rothia, including potential pathogenic species. Overall, saliva of a young child and a senior showed higher bacterial diversity than that of young adults.


A novel p.Ser108LeufsTer15 SOD1 mutation leading to the formation of a premature stop codon in an apparently sporadic ALS patient: insights into the underlying pathomechanisms.
We report an apparently sporadic amyotrophic lateral sclerosis patient carrying a heterozygous novel frameshift SOD1 mutation (p.Ser108LeufsTer15), predicted to cause a premature protein truncation. RT-PCR analysis of SOD1 mRNA and SDS-PAGE/Western blot analysis of PBMC demonstrated that mRNA from the mutant allele is expressed at levels similar to those of the wild-type allele, but the truncated protein is undetectable also in the insoluble fraction and after proteasome inhibition. Accordingly, the dismutation activity in erythrocytes is halved. Thus, the pathogenic mechanism associated with this mutation might be based on an insufficient activity of SOD1 that would make motor neurons more vulnerable to oxidative injury. However, it cannot be excluded that p.Ser108LeufsTer15 SOD1 is present in the nervous tissue and, being less charged and hence having less repulsive forces than the wild-type protein, may trigger toxic mechanisms as a consequence of its propensity to aggregate.


New gene targets of PGC-1α and ERRα co-regulation in C2C12 myotubes.
As a transcriptional coactivator, PGC-1α contributes to the regulation of a broad range of metabolic processes in skeletal muscle health and disease; however, there is limited information about the genes it transcriptionally regulates. To identify new potential gene targets of PGC-1α regulation, mouse C2C12 myotubes were screened by microarray analysis following PGC-1α overexpression. Genes with an mRNA expression of 2.5-fold or more (P < 0.001) were identified. From these, further genes were singled out if they had no previous connection to PGC-1α regulation or characterization in skeletal muscle, or were unannotated with no known function. Following confirmation of their regulation by PGC-1α using qPCR analysis, eight genes were focused on for further investigation (Akr1b10, Rmnd1, 1110008P14Rik, 1700021F05Rik, Mtfp1, Mrm1, Oxnad1 and Cluh). Bioinformatics indicated a number of the genes were linked to a range of metabolic-related functions including fatty acid oxidation, oxido-reductase activity, and mitochondrial remodeling and transport. Treating C2C12 myotubes for 6 h with AICAR, a known activator of AMP kinase and inducer of Pgc-1α gene expression, increased the mRNA levels of both Pgc-1α (P < 0.001) and of Mtfp1, Mrm1, Oxnad1 and Cluh (P < 0.05). Screening of the promoter and intron 1 regions also revealed all genes to contain either a consensus or near consensus response elements for the estrogen-related receptor α (ERRα), a key transcription factor-binding partner of PGC-1α in skeletal muscle. Furthermore, knockdown of endogenous ERRα levels partially or completely blocked the induction of gene expression of all genes by PGC-1α, while each gene was significantly upregulated in the presence of a constitutively active form of ERRα (P < 0.05) except for Akr1b10. These findings provide preliminary evidence for the novel regulation of these genes by PGC-1α and its signaling pathway in skeletal muscle.


Paediatric upper respiratory infections: the role of antibiotics.
To review current clinical evidence for the use of antibiotics in paediatric upper paediatric respiratory infections, repeated PubMed searches using the template algorithm -rhinosinusitis/otitis/ tonsillitis AND ()- with the settings: -Humans; English; All Child 0-18; Clinical trial; Review; Methanalysis; Guideline; Last 10 years- for the following comparators: antibiotic; amoxicillin; clavulanate; penicillin; cephalosporin; macrolide; erythromycin; rokitamycin; clindamycin; trimethoprim-sulfamethoxazole, cefopodoxime, cefdinir, cefuroxime, ceftriaxone. The authors clinical experience in the paediatric allergy unit of a University hospital was also drawn upon. A narrative review was drafted to update paediatricians on the topic. Many paediatric studies and guidelines were retrieved satisfying current evidence-based medicine standards. There are stringent indications for antibiotic use in URTIs. The paediatric use is widespread raising doubts on their appropriate prescription in many countries. Evidence for the efficacy of antibiotic treatment for paediatric URTIs is available and this treatment should be included in individualised patient protocols on the basis of the clinical literature. Caution must be posed at the local level taking in account epidemiologic and microbiologic data to avoid overprescription.


Renal iodide clearance in rabbits.
The purpose of the study was to compare indirect clearance methods based on plasma values and external detection of activity using small skin attached radioactivity detectors with a direct clearance method based on constant infusion and urine collection. The experiments were performed in anesthetized rabbits. The plasma iodide concentration was increased 100 times to prevent thyroid organification of radioactive iodide. [131I]iodide was infused at a constant rate and [125I]iodide was administered iv as a bolus for indirect clearance determination. The ratio between plasma values and external values 125I-activity was constant in all experiments from 15 min after administration of [125I]iodide and throughout the experiment. The results of 7 experiments in 3 rabbits showed a highly significant correlation (r = 0.98) between direct and indirect values within the range of direct clearance values obtained (0.002-0.6 ml.min-1.kg-1. The direct and indirect clearances were measured simultaneously during a 2-h period of steady state. The 95% confidence interval of the mean ratio between direct and indirect clearances was +/- 20% for plasma activity and +/- 26% for externally measured activity. It is concluded that both methods of indirect clearance determination can be applied to rabbits to estimate absolute values of the renal iodide clearance.


Regulation of PI3-kinase/Akt signaling by muscle-enriched microRNA-486.
microRNAs (miRNAs) play key roles in modulating a variety of cellular processes through repression of mRNA targets. In a screen for miRNAs regulated by myocardin-related transcription factor-A (MRTF-A), a coactivator of serum response factor (SRF), we discovered a muscle-enriched miRNA, miR-486, controlled by an alternative promoter within intron 40 of the Ankyrin-1 gene. Transcription of miR-486 is directly controlled by SRF and MRTF-A, as well as by MyoD. Among the most strongly predicted targets of miR-486 are phosphatase and tensin homolog (PTEN) and Foxo1a, which negatively affect phosphoinositide-3-kinase (PI3K)/Akt signaling. Accordingly, PTEN and Foxo1a protein levels are reduced by miR-486 overexpression, which, in turn, enhances PI3K/Akt signaling. Similarly, we show that MRTF-A promotes PI3K/Akt signaling by up-regulating miR-486 expression. Conversely, inhibition of miR-486 expression enhances the expression of PTEN and Foxo1a and dampens signaling through the PI3K/Akt-signaling pathway. Our findings implicate miR-486 as a downstream mediator of the actions of SRF/MRTF-A and MyoD in muscle cells and as a potential modulator of PI3K/Akt signaling.


Rapid immunohistochemistry for cytokeratin in the intraoperative evaluation of sentinel lymph nodes for metastatic breast carcinoma.
The sensitivity and specificity of detecting metastatic breast carcinoma in sentinel lymph nodes using a rapid immunohistochemistry technique was determined and compared with methods currently used at the authors' institution. At the time of intraoperative consultation, after routine diagnostic touch preparations and frozen sections were prepared, 6-microm frozen sections of 72 sentinel lymph nodes from 32 patients with breast carcinoma were placed on plus slides, fixed in cold acetone for 2 or 3 minutes, and stored at -70 degrees C. These sections were immunostained with a prediluted broad-spectrum anticytokeratin monoclonal antibody coupled to an inert polymer with horseradish peroxidase (DAKO EPOS). Slides were ready for interpretation within 16 minutes and were scored as positive, negative, or equivocal for metastatic carcinoma. Results were compared with those of the intraoperative touch preparations and frozen sections and with paraffin-embedded, hematoxylin and eosin-stained, and AE1/AE3 immunostained permanent sections. Fourteen (19%) sentinel lymph nodes were positive for metastatic carcinoma in 13 patients. All methods tested were 100% specific. The rapid immunohistochemistry method was the least sensitive (57% sensitivity) of all methods used to detect metastasis. Routine diagnostic touch preparations, frozen sections, and permanent sections had sensitivities of 69%, 86%, and 100% respectively. In conclusion, this rapid immunohistochemistry method would not be helpful in intraoperative assessment of sentinel lymph nodes in breast cancer patients due to its low sensitivity.


Direct delineation of myocardial infarction without contrast agents using a joint motion feature learning architecture.
Changes in mechanical properties of myocardium caused by a infarction can lead to kinematic abnormalities. This phenomenon has inspired us to develop this work for delineation of myocardial infarction area directly from non-contrast agents cardiac MR imaging sequences. The main contribution of this work is to develop a new joint motion feature learning architecture to efficiently establish direct correspondences between motion features and tissue properties. This architecture consists of three seamless connected function layers: the heart localization layers can automatically crop the region of interest (ROI) sequences involving the left ventricle from the cardiac MR imaging sequences; the motion feature extraction layers, using long short-term memory-recurrent neural networks, a) builds patch-based motion features through local intensity changes between fixed-size patch sequences (cropped from image sequences), and b) uses optical flow techniques to build image-based features through global intensity changes between adjacent images to describe the motion of each pixel; the fully connected discriminative layers can combine two types of motion features together in each pixel and then build the correspondences between motion features and tissue identities (that is, infarct or not) in each pixel. We validated the performance of our framework in 165 cine cardiac MR imaging datasets by comparing to the ground truths manually segmented from delayed Gadolinium-enhanced MR cardiac images by two radiologists with more than 10 years of experience. Our experimental results show that our proposed method has a high and stable accuracy (pixel-level: 95.03%) and consistency (Kappa statistic: 0.91; Dice: 89.87%; RMSE: 0.72 mm; Hausdorff distance: 5.91 mm) compared to manual delineation results. Overall, the advantage of our framework is that it can determine the tissue identity in each pixel from its motion pattern captured by normal cine cardiac MR images, which makes it an attractive tool for the clinical diagnosis of infarction.


Oxytocin attenuates the cocaine-induced exploratory hyperactivity in mice.
The effects of pimozide, a dopamine-receptor blocker and oxytocin, a neurohypophyseal neuropeptide were investigated in mice on the cocaine-induced exploratory hyperactivity. The action of oxytocin on changes of dopaminergic neurotransmission induced by cocaine was also measured. Cocaine-induced exploratory hyperactivity could be blocked by pimozide (1 mg kg-1, s.c.). Oxytocin (0.05-1.0 micrograms) inhibited the cocaine-induced hyperactivity in an U-shaped dose-response manner. In the nucleus accumbens, oxytocin antagonized the increased dopamine disappearance, elicited by cocaine, but not in the nucleus caudatus. The data suggest that oxytocin may influence the behavioural effect of cocaine by modulating dopaminergic neurotransmission in mesolimbic dopaminergic terminal region of the brain.


Experimental and Numerical Studies on the Direct Shear Behavior of Sand-RCA (Recycled Concrete Aggregates) Mixtures with Different Contents of RCA.
Recycled concrete aggregate (RCA) is a typical construction and demolition (C&D) material generated in civil engineering activities and has been widely used as the coarse-grained filler added to sand for roadbed fillings. The effect of RCA content on the mechanical behavior of sand-RCA mixtures is complicated and still not fully understood. To explore the effect of RCA content on the macroscale and microscopic behavior of the sand-RCA mixtures with various RCA contents, laboratory direct shear tests and numerical simulations using the 3D discrete element method were performed. Experimental direct shear tests on sand-RCA mixtures with different contents of RCA were first carried out. Numerical direct shear models were then established to represent the experimental results. The particle shape effect was also considered using a new realistic shape modeling method to model the RCA particles. Good agreement was observed between the DEM simulation and experimental results, verifying the ability of the numerical direct shear models to represent the direct shear behavior of sand-RCA mixtures. The macroscopic responses of both experimental and numerical tests showed that all samples presented an initial hardening followed by a post-peak strain softening. The peak-state friction angles increased with the RCA content for samples under the same vertical stress. The effect of RCA content on the microscopic behavior based on DEM simulation was also found. The microscopic properties of RCA-sand mixtures, such as coordination numbers, PDFs and contact force transformation features, were analyzed and related to the macroscopic results.


Over Expressed TKTL1, CIP-2A, and B-MYB Proteins in Uterine Cervix Epithelium Scrapings as Potential Risk Predictive Biomarkers in HR-HPV-Infected LSIL/ASCUS Patients.
High oncogenic risk human papillomaviruses (HR-HPVs) promote cervical carcinoma development, the fourth most common feminine cancer. A slow oncodevelopmental phase-defined histopathologically as Cervical Intraepithelial Neoplasia (CIN) grades 1-3, or cytologically as Low- or High-grade Squamous Intraepithelial Lesions (LSIL or HSIL)-precedes the malignancy. Cervical carcinoma screenings through HR-HPV genotyping and Pap smears are regularly performed in Western countries. Faulty cytology screening or genotyping or patients' non-compliance with follow-ups can let slip an oncoprogression diagnosis. Novel biomarker tests flanking HR-HPV genotyping and cytology could objectively predict the risk of disease progression thus helping triage LSIL/ASCUS patients. Here, anonymized leftovers of fresh cervical epithelium scrapings from twice (LSIL/ASCUS and HR-HPV DNA)-positive and twice (Pap smear- and HR-HPV DNA)-negative (control) patients in a proteome-preserving solution served to assess the biomarker worth of three cervical carcinoma-related proteins, i.e., B-MYB (or MYBL2), Cancerous Inhibitor of PP2A (CIP-2a), and transketolase-like1 (TKTL1). Leftovers anonymity was strictly kept and storage at -80°C, protein extraction, immunoblotting, and band densitometry were blindly performed. Only after tests completion, the anonymous yet code-corresponding HR-HPV-genotyping and cytology data allowed to assign each sample to the twice-positive or twice-negative group. Descriptive statistics showed that the three proteins levels significantly increased in the twice-positive vs. twice-negative scrapings. Diagnostic ROC curve analysis identified each protein's Optimal Decision Threshold (OTD) showing that TKTL1 and CIP-2a are stronger risk predictive biomarkers (Sensitivity, 0.91-0.93; Specificity, 0.77-0.83) than B-MYB. Logistic Regression coupled with Likelihood-Ratio Tests confirmed that a highly significant relation links increasing TKTL1/CIP-2a/B-MYB protein levels in twice-positive cervical scrapings to the risk of HR-HPV-driven oncoprogression. Finally, a 3 year clinical follow-up showed that 13 patients (50% of total) of the twice-positive group with biomarker values over OTDs compliantly underwent scheduled colposcopy and biopsy. Of these, 11 (i.e., 84.7%) received a positive histological diagnosis, i.e., CIN1 (n = 5; 38.5%) or CIN2/CIN2+ (n = 6; 46,2%). Therefore, TKTL1/CIP-2a/B-MYB protein levels could objectively predict oncoprogression risk in twice (HR-HPV- and Pap smear)-positive women. Further studies will assess the translatability of these findings into clinical settings.


NMR-based techniques in the hit identification and optimisation processes.
In this review, the use of general NMR spectroscopy techniques to detect ligand binding and to monitor enzyme kinetics and inhibition, which appear particularly useful in hit identification and validation, is reiterated. Furthermore, the use of NMR-based strategies for lead optimisations that are based on either iterative derivatisations of an initial core structure or on linking fragments that occupy adjacent pockets in the target's binding site will also be described. Several recent examples will be reported and the use of these techniques in cases when the three dimensional structure of the target protein is known will be discussed.


Effects of weight carrying, exercise and a myo-anabolic supplement on growth and muscle.
Weight training is commonly used by human athletes to increase strength and fitness. This study was performed to examine the effect of weight-carrying and nutritional supplementation on muscle development and growth in young horses. This study examined the effect of weight-carrying and nutritional supplementation on muscle development and growth. Seventeen horses were divided into 3 groups: controls exercised in a free-flow exerciser, a weight group that performed the same exercise, carrying progressively increasing weight up to 45 kg, and a weight-supplement group, that also received a myo-anabolic supplement. Horses were accustomed to handling for 30 days then baseline measurements of weight, body condition score, wither height, hip height, forearm and gaskin circumferences, and cross-sectional area of the longissimus dorsi were taken. Horses were stalled for 108 days then conditioned for 78 days. At the end of conditioning, all measurements were taken again. Supplemented weight-carrying horses increased their lean body tissue, as indicated by greater bodyweight (P<0.05) combined with decreased body condition score (P<0.01). Greater increases in wither height (P = 0.09) and hip height (P<0.01) were seen in weight-carrying horses, probably the result of increased muscle tone. Supplemented weight-carrying horses increased forearm (P<0.01) and gaskin circumferences (P<0.05). It is believed that weight-carrying, when combined with additional dietary protein, enabled greater muscle anabolism. Additional research into the potential additive effect of resistance training and diet are warrented.


Angiotensin-Converting Enzyme 2 (ACE2), Transmembrane Peptidase Serine 2 (TMPRSS2), and Furin Expression Increases in the Lungs of Patients with Idiopathic Pulmonary Fibrosis (IPF) and Lymphangioleiomyomatosis (LAM): Implications for SARS-CoV-2 (COVID-19) Infections.
We previously reported higher ACE2 levels in smokers and patients with COPD. The current study investigates if patients with interstitial lung diseases (ILDs) such as IPF and LAM have elevated ACE2, TMPRSS2, and Furin levels, increasing their risk for SARS-CoV-2 infection and development of COVID-19. Surgically resected lung tissue from IPF, LAM patients, and healthy controls (HC) was immunostained for ACE2, TMPRSS2, and Furin. Percentage ACE2, TMPRSS2, and Furin expression was measured in small airway epithelium (SAE) and alveolar areas using computer-assisted Image-Pro Plus 7.0 software. IPF and LAM tissue was also immunostained for myofibroblast marker α-smooth muscle actin (α-SMA) and growth factor transforming growth factor beta1 (TGF-β1). Compared to HC, ACE2, TMPRSS2 and Furin expression were significantly upregulated in the SAE of IPF (p < 0.01) and LAM (p < 0.001) patients, and in the alveolar areas of IPF (p < 0.001) and LAM (p < 0.01). There was a significant positive correlation between smoking history and ACE2 expression in the IPF cohort for SAE (r = 0.812, p < 0.05) and alveolar areas (r = 0.941, p < 0.01). This, to our knowledge, is the first study to compare ACE2, TMPRSS2, and Furin expression in patients with IPF and LAM compared to HC. Descriptive images show that α-SMA and TGF-β1 increase in the IPF and LAM tissue. Our data suggests that patients with ILDs are at a higher risk of developing severe COVID-19 infection and post-COVID-19 interstitial pulmonary fibrosis. Growth factors secreted by the myofibroblasts, and surrounding tissue could further affect COVID-19 adhesion proteins/cofactors and post-COVID-19 interstitial pulmonary fibrosis. Smoking seems to be the major driving factor in patients with IPF.