Effect of physical activity and fasting on gut and liver proteolysis in the dog.
The aim of this study was to determine how gut and liver protein kinetics adapt to acute exercise in the 18-h-fasted dog (n = 7) and in dogs glycogen depleted by a 42-h fast (n = 8). For this purpose, sampling (artery and portal and hepatic veins) and infusion (vena cava) catheters and Doppler flow probes (portal vein and hepatic artery) were implanted with animals under general anesthesia. At least 16 days later, an experiment, consisting of a 120-min equilibration period, a 30-min basal sampling period, and a 150-min exercise period, was performed. At the start of the equilibration period, a constant rate infusion of [1-13C]leucine was initiated. Gut and liver leucine appearance and disappearance rates were calculated in these studies by combining a novel stable isotopic method and arteriovenous difference methods. In the determination of tissue leucine kinetics the tissue inflow of both alpha-[13C]ketoisocaproic acid and [13C]leucine was taken into account. The results of this study show that 1) the splanchnic bed (liver plus gut) contributes approximately 40% to the whole body proteolytic rate in the basal state and during exercise in dogs fasted for either 18 or 42 h, 2) the contributions of the gut and liver to splanchnic bed proteolysis is about equal in the basal state in both 18- and 42-h-fasted dogs, and 3) exercise in the 18-h-fasted dog leads to a greater emphasis on gut proteolysis and a lesser emphasis on hepatic proteolysis. These studies highlight the important contribution of gut and hepatic proteolysis to whole body proteolysis and the ability of the gut to acutely adapt to changes in physical activity.


Impairment of jejunal absorption rate of carnosine by glycylglycine in man in vivo.
Using a double-lumen tube jejunal perfusion system in vivo, the mutual effects of carnosine (beta-alanyl-L-histidine) and glycylglycine on their respective absorption rates have been studied in six Zambian African adults. Data on the effect of the constituent amino-acids of carnosine on glycylglycine absorption rate have similarly been obtained. The solutions infused in each subject contained (A) carnosine (50 mmol l.-1), (B) carnosine (50 mmol l.-1) and glycylglycine (50 mmol l.-1), (C) glycylglycine (50 mmol l.-1), and (D) glycylglycine (50 mmol l.-1), L-histidine (50 mmol l.-1) and beta-alanine (50 mmol l.-1). Glycylglycine produced a significant impairment in the mean rate of histidine absorption from carnosine (P less than 0-01). However, carnosine did not have a significant effect on the mean rate of glycine absorption from glycylglycine. Mean rate of histidine absorption from solution D was significantly higher than that from solution A (P less than 0-01). Mean rate of glycine absorption from glycylglycine was not significantly different during infusion of solutions B, C, and D. The results are consistent with the concept that carnosine on glycylglycine is probably because the affinity of mechanism; the lack of influence of carnosine on glycylglycine is probably because the affinity of carnosine for the dipeptide uptake mechanism is relatively low. A gross difference has been shown between mean absorption rate of histidine from free L-histidine (solution D) (25-8 mmol h-1) and when it is given in the form of carnosine in the presence of another dipeptide (solution B) (8-7 mmol h-1); that emphasizes the complexity of amino acid and peptide interaction during absorption, which must be important in nutrition.


Organometallic Antitumor Compounds: Ferrocifens as Precursors to Quinone Methides.
The synthesis and chemical oxidation profile of a new generation of ferrocifen derivatives with strong antiproliferative behavior in vitro is reported. In particular, the hydroxypropyl derivative HO(CH2 )3 C(Fc)=C(C6 H4 OH)2 (3 b) exhibited exceptional antiproliferative activity against the cancer cell lines HepG2 and MDA-MB-231 TNBC, with IC50 values of 0.07 and 0.11 μM, respectively. Chemical oxidation of 3 b yielded an unprecedented tetrahydrofuran-substituted quinone methide (QM) via internal cyclization of the hydroxyalkyl chain, whereas the corresponding alkyl analogue CH3 CH2 -C(Fc)=C(C6 H4 OH)2 merely formed a vinyl QM. The ferrocenyl group in 3 b plays a key role, not only as an intramolecular reversible redox "antenna", but also as a stabilized carbenium ion "modulator". The presence of the oxygen heterocycle in 3 b-QM enhances its stability and leads to a unique chemical oxidation profile, thus revealing crucial clues for deciphering its mechanism of action in vivo.


The long-term demographic role of community-based family planning in rural Bangladesh.
Experimental studies demonstrating the effectiveness of nonclinical distribution of contraceptives are typically conducted in settings where contraceptive use is low and unmet need is extensive. Determining the long-term role of active outreach programs after initial demand is met represents an increasingly important policy issue in Asia, where contraceptive prevalence is high and fixed service points are conveniently available. This article examines the long-term rationale for household family planning in Bangladesh-where growing use of contraceptives, rapid fertility decline, and normative change in reproductive preferences are in progress, bringing into question the rationale for large-scale deployment of paid outreach workers. Longitudinal data are analyzed that record outreach encounters and contraceptive use dynamics in a large rural population. Findings demonstrate that outreach has a continuing impact on program effectiveness, even after a decade of household visitation. The policy implications of this finding are reviewed.


[Acute use of adrenergic drugs in asthma and anaphylactic shock in clinical practice].
There are two situations where these products may be used and their justifications are quite different. For anaphylactic shock, the treatment should be aimed at limiting the effects of the acute reduction of volume due to vasoplegia and peripheral diapesis provoked by the liberated histamine. The medullary-suprarenal hormones are the first recourse, efficacious on the alpha and beta receptors. The alpha action is of the excitative type and provokes peripheral vasoconstriction. The beta-action suppresses the contraction of the smooth muscle, but in isolation can facilitate vasodilation. This is why adrenalin is suitable for strict control of arterial pressure: Immediate administration in IM (0.25 to 1 mg), by slow I.V. (dilution 1/10) It is appropriate, where possible to restore the blood volume with a volume expander. Cortico-therapy, both general and by aerosol is used in relays. For asthma: The question is, whether to use beta-2-agonists that have an action on muscle relaxation, on mucosal oedema, also on the secretion of mucus and indirectly on liberation of intermediate substances. Acute use of beta-2-agonists is: Perhaps for a specific purpose, for example to prevent effort-induced asthma: use of a spray is simple and efficacious. Perhaps for a greater and more prolonged use to treat an attack of asthma or a bad state of asthma before hospitalisation.(ABSTRACT TRUNCATED AT 250 WORDS)


Kinetics of salt-induced J-aggregation of cyanine dyes.
The addition of monovalent, divalent, and trivalent metal ions to three anionic ethyl meso-thiacarbocyanine dyes, an ethyl meso-oxacarbocyanine, and an imidacarbocyanine in aqueous solution at room temperature results in the production of J-aggregates within the range of tens to hundreds of seconds. The rate of formation of J-aggregates correlates with the rate of decay of dimers or monomers and is dependent on the type of metal ion, dye structure, and temperature. The rate of formation of J-aggregates increases as the temperature decreases and the dye and salt concentrations increase, and the rate is highest for trivalent ions and smallest for monovalent ions, independent of the type of anion. The time course of formation of J-aggregates is described in most cases by a sigmoidal curve, and the kinetics and mechanism are discussed within the framework of autocatalysis. Computer simulations reveal that the sigmoidal time dependence is transferred to an exponential-like curve by substantially increasing the rate constant for the noncatalytic step. The reaction pathway into J-aggregates can be switched from dimeric ion pairs as the reactant to monomeric ion pairs, when the rate constant for the catalytic step via the monomer becomes larger with respect to that via the dimer.


β-arrestin pathway activation by selective ATR1 agonism promotes calcium influx in podocytes, leading to glomerular damage.
Angiotensin Receptor blockers (ARBs) are the first-line treatment for hypertension; they act by inhibiting signaling through the angiotensin 1 receptor (AT1R). Recently, a novel biased AT1R agonist, TRV120027 (TRV), that selectively activates the β-arrestin cascade and blocks the G-protein-coupled receptor pathway has been proposed as a potential blood pressure medication. Here, we explored the effects of TRV and associated β-arrestin signaling in podocytes, essential cells of the kidney filter. We used human podocyte cell lines to determine β-arrestin's involvement in calcium signaling and cytoskeletal reorganization and Dahl SS rats to investigate the chronic effects of TRV administration on glomerular health. Our experiments indicate that the TRV-activated β-arrestin pathway promotes the rapid elevation of intracellular Ca2+ in a dose-dependent manner. Interestingly, the amplitude of β-arrestin-mediated Ca2+ influx was significantly higher than the response to similar Ang II concentrations. Single-channel analyses show rapid activation of transient receptor potential canonical (TRPC) channels following acute TRV application. Furthermore, the pharmacological blockade of TRPC6 significantly attenuated the β-arrestin-mediated Ca2+ influx. Additionally, prolonged activation of the β-arrestin pathway in podocytes resulted in pathological actin cytoskeleton rearrangements, higher apoptotic cell markers, and augmented glomerular damage. TRV-activated β-arrestin signaling in podocytes may promote TRPC6 channel-mediated Ca2+ influx, foot process effacement, and apoptosis, possibly leading to severe defects in glomerular filtration barrier integrity and kidney health. Under these circumstances, the potential therapeutic application of TRV for hypertension treatment requires further investigation to assess the balance of the benefits versus possible deleterious effects and off-target damage.


VeriStrat: a prognostic and/or predictive biomarker for advanced lung cancer patients?
The role of EGF receptor (EGFR) inhibitors in the treatment of lung cancer without activating EGFR mutations has been a controversial issue, particularly their relative efficacy over the available chemotherapy in the second- and third-line setting. VeriStrat is a serum/plasma proteomic test developed using matrix-assisted laser desorption/ionization methodology, aiming at predicting benefit from EGFR treatment. The VeriStrat algorithm has been interrogated retrospectively and prospectively in samples from randomised trials, such as the PROSE study, confirming the prognostic information associated with the signature. In addition, the test appeared to be predictive of erlotinib impact on survival, as only VeriStrat Good patients benefited from such a treatment. Additional studies should confirm and further define its role in predicting EGFR tyrosine kinase inhibitor benefit, and to establish its better use in terms of clinical efficiency identifying which patients are candidates for the test, at which time on the history of the disease, and lastly at what extra cost.


A truncated tropomyosin-related kinase B receptor, T1, regulates glial cell morphology via Rho GDP dissociation inhibitor 1.
Through tropomyosin-related kinase B (TrkB) receptors, brain-derived neurotrophic factor (BDNF) performs many biological functions such as neural survival, differentiation, and plasticity. T1, an isoform of TrkB receptors that lacks a tyrosine kinase, predominates in the adult mammalian CNS, yet its role remains controversial. In this study, to examine whether T1 transduces a signal and to determine its function, we first performed an affinity purification of T1-binding protein with the T1-specific C-terminal peptide and identified Rho GDP dissociation inhibitor 1 (GDI1), a GDP dissociation inhibitor of Rho small G-proteins, as a signaling protein directly associated with T1. The binding of BDNF to T1 caused Rho GDI1 to dissociate from the C-terminal tail of T1. Astrocytes cultured for 30 d expressed only endogenous T1 among the BDNF receptors. In 30 d cultured astrocytes, Rho GDI1, when dissociated in a BDNF-dependent manner, controlled the activities of the Rho GTPases, which resulted in rapid changes in astrocytic morphology. Furthermore, using 2 d cultured astrocytes that were transfected with T1, a T1 deletion mutant, or cyan fluorescent protein fusion protein of the T1-specific C-terminal sequence, we demonstrated that T1-Rho GDI1 signaling was indispensable for regulating the activities of Rho GTPases and for the subsequent morphological changes among astrocytes. Therefore, these findings indicate that the T1 signaling cascade can alter astrocytic morphology via regulation of Rho GTPase activity.


Motivating older adults through immersive virtual exercise (MOTIVE): A randomized pilot study.
This study aimed to: 1) determine the feasibility of a virtual reality physical activity intervention among older adults and 2) test the preliminary effectiveness of the intervention at increasing physical activity and 3) decreasing depressive symptoms. We included 10 older adults randomized into the Motivating Older Adults Through Immersive Virtual Exercise (MOTIVE) intervention group and 10 randomized into the physical activity education only control group. We analyzed the data using descriptive statistics and linear mixed models, testing the interaction of time and the treatment condition. Participants in the intervention group attended an average of 15 out of the 16 sessions. A total of 90% of MOTIVE intervention group participants "completely agreed" that the intervention was acceptable, (compared to 30% of education control group participants). This study supports testing the effectiveness of the intervention at improving physical activity and depressive symptoms in a larger sample of older adults.


Pore loop-mutated rat KIR6.1 and KIR6.2 suppress KATP current in rat cardiomyocytes.
Cardiomyocytes express mRNA for all major subunits of ATP-sensitive potassium (K(ATP)) channels: KIR6.1, KIR6.2, SUR1A, SUR2A, and SUR2B. It has remained controversial as to whether KIR6.1 may associate with KIR6.2 to form the tetrameric pore of K(ATP) channels in cardiomyocytes. To explore this possibility, cultured rat cardiomyocytes were examined for an inhibition of K(ATP) current by overexpression of pore loop-mutated (inactive) KIR6.x. Bicistronic plasmids were constructed encoding loop-mutated (AFA or SFG for GFG) rat KIR6.x followed by EGFP. In ventricular myocytes, the overexpression of KIR6.1SFG-pIRES(2)-EGFP or KIR6.2AFA-pIRES(2)-EGFP DNA caused, after 72 h, a major decrease of K(ATP) current density of 85.8% and 82.7%, respectively (P < 0.01), relative to EGFP controls (59 +/- 9 pA/pF). In atrial myocytes, overexpression of these pore-mutated KIR6.x by 6.0-fold and 10.6-fold, as assessed by quantitative immunohistochemistry, caused a decrease of K(ATP) current density of 73.7% and 58.5%, respectively (P < 0.01). Expression of wild-type rat KIR6.2 increased the ventricular and atrial K(ATP) current density by 58.3% and 42.9%, respectively (P < 0.01), relative to corresponding EGFP controls, indicating a reserve of SUR to accommodate increased KIR6.x trafficking to the sarcolemma. The results favor the view that KIR6.1 may associate with KIR6.2 to form heterotetrameric pores of native K(ATP) channels in cardiomyocytes.


On recombination among In(2L)t, alpha-Gpdh and Adh in Drosophila melanogaster.
The occurrence and patterns of linkage disequilibrium between an inversion and allozymic loci within it or nearby have been used in attempts to discriminate among different hypotheses for the maintenance of variability. The interpretation of the data on the best-documented case, that of the nonrandom association between In(2L)t and alpha-Gpdh or Adh in the second chromosome of Drosophila melanogaster, has been done on the basis that recombination between alpha-Gpdh and Adh is almost entirely due to the recombination between In(2L)t and the locus within it (alpha-Gpdh), the recombination between the inversion and the nearby locus (Adh) being negligible. In this paper, we show that the pattern of recombination is just the opposite.


Cystobiliary communication in hepatic hydatid cyst: predictors and outcome.
Cystobiliary communication (CBF) with hepatic hydatid disease is responsible for postoperative bile leakage after surgical management. This study aims to detect various predictors of CBF and its outcome after surgical management. This is a retrospective, cohort study of all patients who underwent surgical management for hydatid disease of the liver. Patient data were recorded on an internal web-based registry system supplemented by paper records. Patients were classified into two groups according to the presence of CBF: group (A) patients with CBF and group (B) patients without CBF. There were 123 patients with a hepatic hydatid cyst with a mean age of 39.92±14.59 years. Patients were classified into group (A), 26 patients (21.1%) with CBF, and group (B), 97 patients (78.9%) without CBF. The age group (p=0.04), presence of jaundice (p=0.001), serum glutamic-pyruvic transaminase (SGPT) (p=0.001), cyst size (p=0.0001), and cyst size group (>10 cm) (p=0.0001) were associated with CBF. That cyst size was the only independent predictor of the occurrence of CBF. Intraoperative suturing and the T tube led to complete healing of CBF, and postoperative endoscopic retrograde cholangio-pancreatography (ERCP) and tubal drainage led to a rapid reduction in the bile output and the healing of the fistulas after 9±2.6 days. That cyst size was the only independent predictor for the occurrence of CBF. Management is related to the size of the fistula, the site of the cyst, and the experience of the hepatobiliary surgeon. ERCP is an important option for the management of CBF.


Dental maturity amongst various vertical and sagittal facial patterns.
To compare the mean dental maturity amongst three vertical facial patterns (short, average and long anterior facial heights) and also amongst two sagittal facial patterns (skeletal class I and II) in orthodontic patients. Cross-sectional analytical study. The Aga Khan University Hospital, Karachi, from August to October 2008. Data was retrospectively retrieved from pretreatment orthodontic records of patients who visited the orthodontic clinic. The sample was divided into three vertical groups, namely short, average and long anterior facial types, and two sagittal groups, namely skeletal class I and II. Dental development was assessed using the Demirjian's system. One way ANOVA and independent sample t-test were used to find any difference in dental maturity amongst vertical and sagittal facial patterns, respectively. There was no statistically significant difference in dental maturity amongst the three vertical facial patterns (p=0.111) and also amongst the two sagittal groups (p=0.975). Statistically significant gender dimorphism (p < 0.001) was seen amongst boys and girls for dental maturity with girls being more advanced than boys in their dental development. Dental maturation in the three vertical and the two sagittal groups showed no statistically significant difference in our studied population sample and thus need no variations in treatment initiation timings. However, girls should be treated earlier as they are dentally more advanced than boys.


Is overweight a risk factor for sports injuries in children, adolescents, and young adults?
Physical activity and sports participation are promoted to counteract the increased prevalence of overweight and obesity in children and young adults. Both high body mass index and physical activity level have been associated with an increased risk of sports injuries. The objective is to determine the relationship between sports injuries and overweight in sports participants (4-24 years), taking physical activity into account. Data were obtained from the 2006-2011 "Injuries and Physical Activity in the Netherlands" survey. Analyses were based on a representative sample of 3846 sports participants (4-24 years). Univariate and multiple logistic regression analyses were applied to investigate the association between sports injury and weight status. Of all the sports participants, 14.7% were overweight. Compared with normal-weight sports participants, the odds of sustaining a sports injury was 0.73 [confidence interval (CI): 0.53-1.00, P = 0.050] for overweight sports participants; the odds for underweight sports participants was 0.80 (CI: 0.56-1.15, P = 0.226). There is some evidence that overweight sports participants (4-24 years) do not have an increased injury risk compared with normal-weight sports participants, even when the level of physical activity is taken into account. Additional research is recommended regarding overweight people who start to participate in a physically active lifestyle.


Role of glycosidases in human ovarian carcinoma cell mediated degradation of subendothelial extracellular matrix.
Penetration of the extracellular matrix (ECM) by tumor cells, an event which occurs at various stages of the metastatic process, involves tumor cell glycosidase mediated hydrolysis of proteoglycans (PG). Recently, we observed that human ovarian carcinoma cell lines (HOCC) derived from primary tumors, peritoneal effusions, and distant metastases possess a varying ability to degrade radiolabeled PG of the ECM, while normal cells (human mesothelial cells or ovarian fibroblasts) fail to do so. To determine whether a quantitative relationship exists between glycosidase activity and degradation of ECM, both intracellular and extracellular glycosidase activities were measured for HOCC and normal cell lines. No relationship was found between intracellular glycosidase activities and the ability of cells to degrade ECM. However, a correlation was observed between extracellular or secretory glycosidase activities and HOCC mediated ECM degradation. In particular, a 5-8-fold increase, as compared to normal cells, was observed for HOCC extracellular beta-N-acetylglucosaminidase (EC 184.108.40.206) activity. The accumulation or secretion of this enzyme from HOCC into culture medium was found to be time dependent and not related to intracellular levels. Purified hexosaminidase derived from invasive HOCC was able to hydrolyze [3H]-glucosamine radiolabeled ECM (up to 30% radiolabel) and resulted in the cumulative release of free [3H]-N-acetylglucosamine. This enzyme mediated hydrolysis could be completely prevented with 2-acetamido-2-deoxy-1,5-D-gluconolactone, a competitive inhibitor (Ki 10(-6) M). Finally, HOCC mediated degradation of radiolabeled ECM was discerned to be dependent upon active hexosaminidase action, since tumor cell mediated degradation of ECM could be inhibited by up to 60% in the presence of this synthetic competitive inhibitor. In summary, these studies indicate a strong association between HOCC solubilization of glycoconjugates present in the ECM and extracellular levels of hexosaminidase.


Antibodies to S2 domain of SARS-CoV-2 spike protein in Moderna mRNA vaccinated subjects sustain antibody-dependent NK cell-mediated cell cytotoxicity against Omicron BA.1.
Vaccination with the primary two-dose series of SARS-CoV-2 mRNA protects against infection with the ancestral strain, and limits the presentation of severe disease after re-infection by multiple variants of concern (VOC), including Omicron, despite the lack of a strong neutralizing response to these variants. We compared antibody responses in serum samples collected from mRNA-1273 (Moderna) vaccinated subjects to identify mechanisms of immune escape and cross-protection. Using pseudovirus constructs containing domain-specific amino acid changes representative of Omicron BA.1, combined with domain competition and RBD-antibody depletion, we showed that RBD antibodies were primarily responsible for virus neutralization and variant escape. Antibodies to NTD played a less significant role in antibody neutralization but acted along with RBD to enhance neutralization. S2 of Omicron BA.1 had no impact on neutralization escape, suggesting it is a less critical domain for antibody neutralization; however, it was as capable as S1 at eliciting IgG3 responses and NK-cell mediated, antibody-dependent cell cytotoxicity (ADCC). Antibody neutralization and ADCC activities to RBD, NTD, and S1 were all prone to BA.1 escape. In contrast, ADCC activities to S2 resisted BA.1 escape. In conclusion, S2 antibodies showed potent ADCC function and resisted Omicron BA.1 escape, suggesting that S2 contributes to cross-protection against Omicron BA.1. In line with its conserved nature, S2 may hold promise as a vaccine target against future variants of SARS-CoV-2.


[Antitumor and toxic effects of amotin].
An indole alkaloid isolated from Catharanthus roseus in the Soviet Union and named amotin differs in terpenoid moiety structure of the indole part of the molecule from the foreign preparation vinblastine and its Soviet analog rosevine. In experimental study amotin has exhibited a high antileukemic activity which was more expressed than that of vinblastine. Tolerant doses of amotin cause moderate reversible morphological alterations in hematopoietic organs, gastrointestinal mucosa, liver, kidney, adrenals, testes, ovaries.


Morphologic characteristics of palatopharyngeal muscle.
In an effort to clarify the morphologic characteristics of the palatopharyngeal muscle, we examined its origin, insertion, and positional relationship with other muscles. We found that the origin of the palatopharyngeal muscle was both the oral and the nasal side of the soft palate; it was also attached to both the palatal aponeurosis and the soft palate median. However, in some cases the muscle originated on the nasal side was lacked. When the palatopharyngeal muscle originated from both the oral and the nasal side, it traveled through its insertion via the levator muscle of the palatine velum. This insertion was seen in a wide area and could be divided into three parts: the pharynx anterior, central, and posterior walls. In the central pharyngeal wall, insertion into the pharyngeal aponeurosis, inferior constrictor pharyngeal muscle, and esophagus were observed. The present results suggest that the palatopharyngeal muscle has a close positional relationship with the levator and tensor muscles of the palatine velum, the pharyngeal constrictor muscles, and the esophagus.


Preparation, characterization, and stability of liposome-based formulations of mitoxantrone.
The preparation, characterization, and stability of lyophilized liposome-based formulation of mitoxantrone was investigated. Mitoxantrone was entrapped inside small, unilamellar liposomes composed of dioleoylphosphocholine (DOPC), cholesterol, and cardiolipin. The mean vesicle size and drug entrapment efficiency of the liposomes were approximately 150 nm and approximately 99%, respectively. Less than 1% of drug was lost and mean vesicle size remained unchanged after sterile filtration. The pre-lyophilized (pre-lyo) formulations were characterized by a differential scanning calorimetric (DSC) method. Results showed that the glass transition temperatures (Tg) increased as the molar ratios of sucrose:lipid and trehalose:lipid in the formulations were increased. The maximum Tg' of the pre-lyo formulations containing 10:1 sucrose:lipid and trehalose:lipid molar ratios were -37C and -41C, respectively. After reconstitution of the lyophilized cake of the sucrose-containing formulation, the mean vesicle size was comparable to pre-lyo liposome size. In vitro release studies showed that less than 2% of mitoxantrone was released after an extensive dialysis against phosphate buffered saline (PBS) at 37C, indicating that the mitoxantrone was highly associated and retained inside the liposomes. Short-term stability studies of the sucrose-containing formulations revealed that the reconstituted and eight-fold diluted formulations were stable for up to 8 hours at room temperature. Long-term stability studies of lyophilized liposomal mitoxantrone showed that the lyophilized formulation was stable for up to 13 months after storage at refrigerated condition.


All-optical pulse compression of broadband microwave signal based on stimulated Brillouin scattering.
Pulse compression processing based on stimulated Brillouin scattering (SBS) in an optical fiber is theoretically and experimentally demonstrated. Broadband microwave signal is electro-optically modulated onto the pump lightwave that is launched into one end of the fiber. Acoustic wave in the fiber inherits the amplitude and phase information of the pump lightwave and thus the coupling between the acoustic wave and pump lightwave leads to the auto-correlated process of the pump lightwave as well as the modulated microwave signal. Derivation of the SBS coupling equations shows that the short-pulse probe lightwave amplified by the pump lightwave possesses the nature of auto-correlation formula. All-optical pulse compression of the broadband microwave signal is implemented after a subtraction between the detected probe pulse with and without SBS. A proof-of-concept experiment is carried out. The pulse compression of a linear frequency-modulated microwave signal with 1 GHz sweep range at the carrier frequency of 4.3 GHz is successfully realized, which well matches the theoretical analysis.


Multiplex optical biosensors based on multi-pinhole interferometry.
The application of new sensor technologies for frequent biomarker monitoring in combination with the leverage of artificial intelligence has great potential to improve the design and safety of health care. With current research efforts, the screening of tens of biomarkers at the point of care and immediate adjustment of therapy is coming within reach. Here we introduce an optical multiplexing approach based on multi-pinhole interference providing inherent differential referencing between a multitude of measurement fields on a surface. A theoretical study of an 11-plex and a 54-plex design is complemented with the experimental demonstration of the technique for a 3-field refractive index measurements and detection of human α-thrombin.


Investigation of the Effects of Pulse-Atomic Force Nanolithography Parameters on 2.5D Nanostructures' Morphology.
In recent years, Atomic Force Microscope (AFM)-based nanolithography techniques have emerged as a very powerful approach for the machining of countless types of nanostructures. However, the conventional AFM-based nanolithography methods suffer from low efficiency, low rate of patterning, and high complexity of execution. In this frame, we first developed an easy and effective nanopatterning technique, termed Pulse-Atomic Force Lithography (P-AFL), with which we were able to pattern 2.5D nanogrooves on a thin polymer layer. Indeed, for the first time, we patterned nanogrooves with either constant or varying depth profiles, with sub-nanometre resolution, high accuracy, and reproducibility. In this paper, we present the results on the investigation of the effects of P-AFL parameters on 2.5D nanostructures' morphology. We considered three main P-AFL parameters, i.e., the pulse's amplitude (setpoint), the pulses' width, and the distance between the following indentations (step), and we patterned arrays of grooves after a precise and well-established variation of the aforementioned parameters. Optimizing the nanolithography process, in terms of patterning time and nanostructures quality, we realized unconventional shape nanostructures with high accuracy and fidelity. Finally, a scanning electron microscope was used to confirm that P-AFL does not induce any damage on AFM tips used to pattern the nanostructures.


Psychosis in behavioral variant frontotemporal dementia.
Dementia is generally characterized by cognitive impairment that can be accompanied by psychotic symptoms; for example, visual hallucinations are a core feature of dementia with Lewy bodies, and delusions are often seen in Alzheimer's disease. However, for behavioral variant of frontotemporal dementia (bvFTD), studies on the broad spectrum of psychotic symptoms are still lacking. The aim of this study was to systematically and prospectively subtype the wide spectrum of psychotic symptoms in probable and definite bvFTD. In this study, a commonly used and validated clinical scale that quantifies the broad spectrum of psychotic symptoms (Positive and Negative Symptom Scale) was used in patients with probable and definite bvFTD (n=22) and with a primary psychiatric disorder (n=35) in a late-onset frontal lobe cohort. Median symptom duration was 2.8 years, and the patients were prospectively followed for 2 years. In total, 22.7% of bvFTD patients suffered from delusions, hallucinatory behavior, and suspiciousness, although the majority of the patients exhibited negative psychotic symptoms such as social and emotional withdrawal and blunted affect (95.5%) and formal thought disorders (81.8%). "Difficulty in abstract thinking" and "stereotypical thinking" (formal thought disorders) differentiated bvFTD from psychiatric disorders. The combined predictors difficulty in abstract thinking, stereotypical thinking, "anxiety", "guilt feelings," and "tension" explained 75.4% of variance in the diagnosis of bvFTD versus psychiatric diagnoses (P<0.001). Delusions, hallucinatory behavior, and suspiciousness were present in one-fifth of bvFTD patients, whereas negative psychotic symptoms such as social and emotional withdrawal, blunted affect, and formal thought disorders were more frequently present. This suggests that negative psychotic symptoms and formal thought disorders have an important role in the psychiatric misdiagnosis in bvFTD; misdiagnosis in bvFTD might be reduced by systematically exploring the broad spectrum of psychiatric symptoms.


Proteomic pattern of plasma in off-pump coronary artery bypass surgery.
The aim of the present study was to identify, using proteomics, changes in the expression of proteins and protein isoforms mainly associated with inflammation and vascular damage in the coronary sinus of patients undergoing off-pump coronary artery bypass (OPCAB) surgery. Plasma from the coronary sinus of 18 patients undergoing OPCAB was obtained before the ischemic procedure and after revascularization of the left anterior descending artery (LAD) with an internal thoracic artery graft. The mean LAD ischemic time was 11.1+/-0.7 min. For the proteomic study, two-dimensional gel electrophoresis was performed. A significant decrease in the expression of three fibrinogen gamma-chain (FGC) isoforms, three vitamin D-binding protein (DBP) isoforms, six haptoglobin (HPT) isoforms, two apolipoprotein (apo)-AI isoforms, one ceruloplasmin (CP) isoform and apoAIV was found after revascularization. Before anastomosis, negative correlations between blood flow and FGC isoforms 2 and 3 and positive correlations between blood flow and apoAI isoform 5 were observed. After anastomosis, we observed positive correlations between haptoglobin isoform 3 and DBP isoform 2 and blood flow. Before anastomosis, positive correlation between DBP isoform 2 and troponin I was observed. After LAD grafting, positive correlations between troponin I and HPT isoform 6, CP isoform 1 and apoAI isoforms 2 and 4 were observed. After the procedure, positive correlations between creatine kinase-MB and coronary sinus expression of FGC isoforms 1, 2 and 3 and HPT isoforms 1 and 2 were also observed. conclusions: In blood from coronary sinus the expression of a number of proteins and protein isoforms associated with inflammation and vascular protection was modified after OPCAB.


The value of transesophageal echography in the clinical staging of lung cancer.
The noninvasive preoperative assessment of lung cancer is currently based on CT scan. This technique is used to find out the local extension of the tumor and to identify the presence of mediastinal adenopathies by distinguishing as accurate as possible between benign and metastatic adenopathies according to their size. When CT scan suggests the presence of suspected mediastinal lymph nodes, invasive diagnostic methods are indicated to confirm whether the lymph nodes are affected (transbronchial needle aspiration, mediastinoscopy, mediastinotomy and thoracoscopy). Transesophageal echography (TEE) was first used in the 80's as a diagnostic guide in the clinical classification of gastrointestinal tumors and may be of help in the mediastinal evaluation of lung cancer as well. Several studies have demonstrated the value of TEE in detecting mediastinal adenopathies and mediastinal structures involvement (great vessels and mediastinal organs) by assessing the direct growth of the primary tumor which is crucial in the surgical decision. To distinguish benign from metastatic nodes TEE, unlike CT scan, does not consider their size only but other issues as shape, definition of edges and echographic structure. Generally speaking, TEE detects more adenopathies than CT scan but not in all mediastinal stations since it is limited by the presence of air; thus, adenopathies located in the upper mediastinum, mainly on the right side, are usually missed by TEE. The areas more clearly visualized are the subcarinal, paraesophageal, tracheobronchial and hilar ones and mainly those located in the aortopulmonary window. Currently, TEE can be used also to guide a thin needle toward the lymph nodes and to perform its aspiration biopsy. In conclusion, transesophageal and/or endobronchial echography are developing promising diagnostic methods which provide additional information that can influence the clinical decision making process and the prognosis. They are well tolerated by the patients but time is needed to find out their role. Lastly, their usefulness will depend also on their feasibility since an expensive equipment and specifically well trained staff are needed.


Different actions of gamma-hydroxybutyrate: a critical outlook.
Gamma-hydroxybutyrate acid (GHB) is a naturally occurring analog of GABA in the mammalian brain and can be therapeutically used for basic sedation in intensive care units. Although its application is discussed controversially, GHB is suspected to protect neuronal tissue against ischemic damage. GHB was tested for an acute effect on electrophysiologic parameters of guinea pig hippocampal tissues exposed to ischemic conditions. With application of 0.5 mM GHB, an acute protective effect was observed. The aim of the present paper is to discuss our experimental results as well as pathophysiological mechanisms of GHB and its clinical applicability.


Low-frequency respiratory rhythms in infants during the first six months of life.
The aim of the study was to investigate characteristics of low-frequency components in respiration. Sixteen healthy term infants were examined from the first day up to the 6th month of life. The respirogram, instantaneous respiratory frequency and respiratory amplitude of undisturbed segments of quiet sleep phases and periodic breathing (PB) were analysed via fast Fourier transformation. The peak frequency (PF) in the low-frequency range (0.04-0.2 Hz) was determined. PF for PB ranged from 0.056 to 0.1 Hz. Further, low-frequency rhythms (LFR) of the respirogram, which were stable during the recordings as well as during development, were found ranging from 0.045 to 0.067 Hz. The LFR of the respirogram is correlated with rhythmic changes in the relationship between inspiratory and expiratory amplitudes. The frequency of the LFR was significantly lower than that of the PB. The data indicate that LFR and PB are low-frequency respiratory rhythms which are separately controlled and perform independently.


Environmental Heterogeneity Leads to Spatial Differences in Genetic Diversity and Demographic Structure of Acer caudatifolium.
Under climate fluctuation, species dispersal may be disturbed by terrain and local climate, resulting in uneven spatial-genetic structure. In addition, organisms at different latitudes may be differentially susceptible to climate change. Here, we tracked the seed dispersal of Acer caudatifolium using chloroplast DNA to explore the relationships of terrain and local climate heterogeneity with range shifts and demography in Taiwan. Our results showed that the extant populations have shifted upward and northward to the mountains since the Last Glacial Maximum. The distributional upshift of A. caudatifolium is in contrast to the downward expansion of its closest relative in Taiwan, A. morrisonense. The northern populations of A. caudatifolium have acquired multiple-source chlorotypes and harbor high genetic diversity. However, effective gene flow between the north and south is interrupted by topography, geographic distance, north-south differences in October rainfall, and other climate heterogeneities, blocking southward genetic rescue. In addition, winter monsoon-driven rainfall may cause regional differences in the phenological schedule, resulting in adaptive effects on the timing of range shift and the genetic draft of chlorotype distribution. Terrain, distance, and local climate also differentiate the northernmost populations from the others, supporting the previous taxonomic treatment of Acer kawakamii var. taitonmontanum as an independent variety.


Effects of Valvular Heart Disease on Clinical Outcomes in Sarcoidosis.
Systemic sarcoidosis can lead to heart failure, conduction abnormalities and ventricular arrhythmias although data on concomitant valvular heart disease (VHD) is limited. We reported the prevalence and outcomes of VHD in systemic sarcoidosis. A retrospective cohort study was conducted using National Inpatient Sample between 2016 and 2020 with respective ICD-10-CM codes. 406,315 patients were hospitalized with sarcoidosis, out of which 20,570 had comorbid VHD (5.1%). Mitral disease was most common (2.5%), followed by aortic, and tricuspid disease. Tricuspid disease was associated with increased mortality in sarcoidosis (OR 1.6, 95% CI, 1.1-2.6, P = 0.04), while aortic disease was associated with higher mortality in only 31-50 years age cohort. Patients with sarcoidosis and VHD have higher hospitalization charges and lower or similar valvular intervention rates than those without sarcoidosis. VHD has a prevalence of 5% in sarcoidosis, predominantly affecting mitral and aortic valves. Underlying VHD is associated with worse outcomes in sarcoidosis.


Development and characterization of a model of liver metastasis using human colon cancer HCT-116 cells.
In order to develop a model of liver metastasis of human gastrointestinal cancer cells, we examined the potential of 10 human colon and stomach cancer cell lines (HT-29, WiDr, HCT-116, HCT-15, HCC-2998, MKN7, MKN28, MKN45, MKN74 and St-4) to form liver metastases in nude mice. Among the cell lines, HCT-116 cells consistently formed gross liver metastases when injected into the spleens of nude mice. In contrast, other human colon and stomach cancer cells produced little or no liver metastasis. In order to analyze the high metastatic potential of HCT-116 cells, the adhesion potential was compared between HCT-116 cells and the other colon cancer cell lines. HCT-116 cells showed more efficient adhesion to fibronectin (FN) than other cells. Furthermore, FN enhanced haptotaxis of HCT-116 cells, but not of other colon cancer cells. The high adhesion potential to FN and enhanced haptotaxis may contribute, at least in part, to the high metastatic potential of HCT-116. To assess the value of this newly developed model of liver metastasis, we compared the ability of four anticancer drugs (fluorouracil, doxifluridine, paclitaxel and irinotecan) to inhibit the formation of liver metastases. Paclitaxel and irinotecan showed strong inhibition of liver metastasis but fluorouracil and doxifluridine showed only slight inhibition. Therefore, this model of metastasis may be useful for screening anti-liver metastatic reagents. These results indicate that the HCT-116 liver-metastasis model should be useful for analyzing the molecular mechanism of liver metastasis and for evaluating new anti-liver metastatic drugs.


Detection methods targeting the positive- and negative-sense RNA transcripts from plus-stranded RNA viruses.
The largest group of viruses in the Baltimore classification system comprises viruses with a positive-sense, single-stranded RNA genome. Once the viral genome is released into the cytoplasm of a specific host cell following virus entry, it functions directly as an mRNA, and the virus-encoded proteins that are essential for genome replication are produced by the translation apparatus of the host cell. The positive-sense genome is replicated in two stages, initially the positive strand is copied to make a negative-sense RNA, which then functions as the template for transcription of many new positive-sense genomes. Virus infections can be detected at different stages throughout the infection cycle for diagnostic and scientific purposes. Here, the advantages and disadvantages of some of the relevant methods for genome detection will be briefly reviewed with special emphasis on techniques allowing strand-specific RNA detection. Furthermore, tools of the future are considered.


Long non-coding RNA FAM83H-AS1 acts as a potential oncogenic driver in human ovarian cancer.
Ovarian cancer (OC) is one of the most aggressive women cancers with increasing incidence and mortality rates worldwide. Long non-coding RNAs (lncRNAs) could as major players in OC process. Although FAM83H antisense RNA1 (FAM83H-AS1) is demonstrated play an important roles in a many cancers, the detailed function and mechanism has not been reported in OC. We integrated multiple kinds of bioinformatics approaches and experiments validated method to evaluate functions of FAM83H-AS1 in OC. Some differential expressed lncRNAs were identified between OC and normal control tissues. FAM83H-AS1 was one of most differentially expressed lncRNAs and up-regulated in multiple cancer types. Specially, expression of FAM83H-AS1 was higher in OC and showed difference in diverse stages. High FAM83H-AS1 expression is associated with worse pan-cancer and OC outcomes. FAM83H-AS1-centric network including lncRNA-miRNA, lncRNA-protein and lncRNA-mRNA ceRNA network were constructed to infer the function and mechanism of FAM83H-AS1. There were two methylation sites including cg01399317 and cg20519035 located at FAM83H-AS1. The methylation level of cg01399317 was correlated with gene expression of FAM83H-AS1. The expression level of FAM83H-AS1 was correlated with infiltration level of immune cell including macrophage, neutrphil and dendritic cell in OC patients. Lastly, qRT-PCR showed that the expression of FAM83H-AS1 was higher in OC tissues than normal control tissues. Collectively, these results indicated that FAM83H-AS1 may act as an oncogenic driver and it may be a potential therapy target in OC.


Staphylococcus aureus PBP4 is essential for beta-lactam resistance in community-acquired methicillin-resistant strains.
Recent cases of infections caused by community-acquired methicillin-resistant Staphylococcus aureus (MRSA) (CA-MRSA) strains in healthy individuals have raised concerns worldwide. CA-MRSA strains differ from hospital-acquired MRSAs by virtue of their genomic background and increased virulence in animal models. Here, we show that in two common CA-MRSA isolates, USA300 and MW2 (USA400), a loss of penicillin binding protein 4 (PBP4) is sufficient to cause a 16-fold reduction in oxacillin and nafcillin resistance, thus demonstrating that mecA, encoding PBP2A, is not the sole determinant of methicillin resistance in CA-MRSA. The loss of PBP4 was also found to severely affect the transcription of PBP2 in cells after challenge with oxacillin, thus leading to a significant decrease in peptidoglycan cross-linking. Autolysis, which is commonly associated with the killing mechanism of penicillin and beta-lactams, does not play a role in the reduced resistance phenotype associated with the loss of PBP4. We also showed that cefoxitin, a semisynthetic beta-lactam that binds irreversibly to PBP4, is synergistic with oxacillin in killing CA-MRSA strains, including clinical CA-MRSA isolates. Thus, PBP4 represents a major target for drug rediscovery against CA-MRSA, and a combination of cefoxitin and synthetic penicillins may be an effective therapy for CA-MRSA infections.


Genome sequence of freshwater sediment isolated fungus Westerdykella aurantiaca strain NNIBRFG27121.
We sequenced the genome of Westerdykella aurantiaca NNIBRFG27121 strain isolated from the wetland of Maehwamarum Habitat in Korea. The final assembly consisted of six scaffolds with a size of 31.96 Mb and an N50 of 8,770,400 bp. This genome will help in comparing species within the Westerdykella genus.


Atmospheric aerosols investigated by inversion of experimental transmittance data.
The Twomey-Chahine inversion algorithm is applied to experimental atmospheric transmittance data in the 0.4-2.4-microm wavelength range and atmospheric aerosol size distributions deduced. The conditions for successful inversion of transmittance data are investigated in numerical experiments, and it is shown that too small a wavelength range results in a Junge-type distribution in all cases and that noise in the measurements in excess of 4-5% results in inversion artifacts.


Sequence-Encoded Differences in Phase Separation Enable Formation of Resilin-like Polypeptide-Based Microstructured Hydrogels.
Microstructured hydrogels are promising platforms to mimic structural and compositional heterogeneities of the native extracellular matrix (ECM). The current state-of-the-art soft matter patterning techniques for generating ECM mimics can be limited owing to their reliance on specialized equipment and multiple time- and energy-intensive steps. Here, a photocross-linking methodology that traps various morphologies of phase-separated multicomponent formulations of compositionally distinct resilin-like polypeptides (RLPs) is reported. Turbidimetry and quantitative 1H NMR spectroscopy were utilized to investigate the sequence-dependent liquid-liquid phase separation of multicomponent solutions of RLPs. Differences between the intermolecular interactions of two different photocross-linkable RLPs and a phase-separating templating RLP were exploited for producing microstructured hydrogels with tunable control over pore diameters (ranging from 1.5 to 150 μm) and shear storage moduli (ranging from 0.2 to 5 kPa). The culture of human mesenchymal stem cells demonstrated high viability and attachment on microstructured hydrogels, suggesting their potential for developing customizable platforms for regenerative medicine applications.


CCL3 contributes to secondary damage after spinal cord injury.
Secondary damage after spinal cord injury (SCI) is characterized by a cascade of events including hemorrhage, apoptosis, oxidative stress, and inflammation which increase the lesion size which can influence the functional impairment. Thus, identifying specific mechanisms attributed to secondary injury is critical in minimizing tissue damage and improving neurological outcome. In this work, we are investigating the role of CCL3 (macrophage inflammatory protein 1-α, MIP-1α), a chemokine involved in the recruitment of inflammatory cells, which plays an important role in inflammatory conditions of the central and peripheral nervous system. A mouse model of lower thoracic (T11) spinal cord contusion injury was used. We assessed expression levels of CCL3 and its receptors on the mRNA and protein level and analyzed changes in locomotor recovery and the inflammatory response in the injured spinal cord of wild-type and CCL3-/- mice. The expression of CCL3 and its receptors was increased after thoracic contusion SCI in mice. We then examined the role of CCL3 after SCI and its direct influence on the inflammatory response, locomotor recovery and lesion size using CCL3-/- mice. CCL3-/- mice showed mild but significant improvement of locomotor recovery, a smaller lesion size and reduced neuronal damage compared to wild-type controls. In addition, neutrophil numbers as well as the pro-inflammatory cytokines and chemokines, known to play a deleterious role after SCI, were markedly reduced in the absence of CCL3. We have identified CCL3 as a potential target to modulate the inflammatory response and secondary damage after SCI. Collectively, this study shows that CCL3 contributes to progressive tissue damage and functional impairment during secondary injury after SCI.


[Correlation of MRI apparent diffusion coeffcient with molecular marker Ki-67 in gastric cancer].
To study the correlation between MRI apparent diffusion coefficient (ADC) and expression of Ki-67 in gastric cancers, and to investigate the application of ADC value in diagnosing the malignance of gastric cancer. A retrospective cohort analysis was performed on 87 gastric cancer patients who received MRI examination and radical resection at Zhejiang Provincial Hospital of Traditional Chinese Medicine from November 2014 to August 2015. All the postoperative resected samples were confirmed as gastric cancer. Preoperative MRI examination was performed by using Siemens 3.0-T Verio MRI with following parameters: section thickness 3 mm, gap 1 mm, matrix 182×320, field of view 40 cm. Plain scan was followed by T1-weighted fat suppression technique VIBE 3D(TR3.92/TE1.39,90degree) scans at arterial phase (the 30th second), portal venous phase (the 60th second), lag period (the 90th second), axial planes and coronal planes (the 180th second), and sagittal planes (the 210th second), respectively. ADC value of tumor was measured at b-factor of 800 s/mm2 and ADC map was generated from DWI data on the work station. The expression of Ki-67 in cancer tissue was detected by routine immunohistochemical (SP) staining after surgery. Correlation between ADC value and the expression of Ki-67 in gastric cancer was analyzed. Irregular thickening of the gastric wall and inhomogeneous enhancement of the tumor after injection of the contrast material appeared in gastric cancer. Gastric cancer tissue presented hyperintensity and normal gastric wall presented isointensity in DWI image (b=800 s/mm2). Compared with normal gastric tissue, mean ADC value of gastric cancer tissue was significant lower [(1.114±0.265)×10-3 mm2/s vs. (2.032±0.202)×10-3 mm2/s, t=26.209, P=0.000]. The ADC values of high-middle differentiation group, middle-low differentiation group, low differentiation group and signet ring cell carcinoma/mucinous adenocarcinoma group were (1.347±0.234)×10-3 mm2/s, (1.179±0.257)×10-3 mm2/s, (0.996±0.185)×10-3 mm2/s and (1.082±0.230)×10-3 mm2/s, respectively. The difference of mean ADC value among different tumor stages was significant(F=8.498, P=0.000). Along with the Ki-67 expression up-regulated, the ADC value decreased in cancer tissue. The Ki-67 expressions in cancer tissue was negatively correlated with cancer ADC values (r=-0.570, P=0.000). Furthermore, negative correlations of Ki-67 expressions with ADC values of high-middle differentiation group (r=-0.627, P=0.016), low differentiation group (r=-0.787, P=0.000) and signet ring cell carcinoma/mucinous adenocarcinoma group (r=-0.792, P=0.000) were observed respectively, while Ki-67 expression was not correlated with ADC value of middle-low differentiation group. The ADC value of gastric cancer can reflect the level of tumor differentiation, and is negatively correlated with Ki-67 expression in cancer tissues.


Comparing Auditory Brain Stem Responses and Transient Otoacoustic Emissions in Premature Infants with Auditory Developmental Delay: Evidence of Temporary Auditory Neuropathy.
Premature birth causes some permanent or temporary abnormalities in the hearing system of the newborn. Inadequate development of the central auditory nervous system and balance, as well as the delay in the formation of the nerve myelin, can be the cause of many hearing disorders, including permanent or temporary auditory neuropathy spectrum disorder (ANSD). The present study aims to identify and understand developmental delay disorder in the hearing system of infants and investigate the possibility of temporary auditory neuropathy in infants. In this comparative analytical study, twenty premature infants were randomly selected for hearing tests using auditory brainstem response and transient otoacoustic emissions at the time of discharge and three months after the first evaluation. The different components of these tests were analyzed and compared before and after developing the auditory system. The OAEs test showed a signal-to-noise ratio above six dB with appropriate amplitudes in all infants. The grand average waveform of the ABR showed a significant difference between the amplitudes of waves III and V before and after maturation in both ears (p<0.05). In addition, the absolute latency of waves, specifically III and V, showed a significant difference between the two assessment times (0.05). The present study confirmed the occurrence of temporary ANSD or delayed maturation in premature infants following the lack of complete growth and myelination of auditory nerve fibers. There is a need to determine the hearing status of premature infants by frequent examinations and prevent any unnecessary prescription of amplifications.


N-Glycosylation influences transport, but not cellular trafficking, of a neuronal amino acid transporter SNAT1.
SNAT1 is a system N/A neutral amino acid transporter that primarily expresses in neurons and mediates the transport of l-glutamine (Gln). Gln is an important amino acid involved in multiple cellular functions and also is a precursor for neurotransmitters, glutamate and GABA. In the present study, we demonstrated that SNAT1 is an N-glycoprotein expressed in neurons. We identified three glycosylation sites at asparagine residues 251, 257 and 310 in SNAT1 protein, and that the first two are the primary sites. The biotinylation and confocal immunofluorescence analysis showed that the glycosylation-impaired mutants and deglycosylated SNAT1 were equally capable of expressing on the cell surface. However, l-Gln and 3H-labeled methyl amino isobutyrate (MeAIB) was significantly compromised in N-glycosylation-impaired mutants and deglycosylated SNAT1 when compared with the wild-type control. Taken together, these results suggest that SNAT1 is an N-glycosylated protein with three de novo glycosylation sites and N-glycosylation of SNAT1 may play an important role in the transport of substrates across the cell membrane.


[The perspectives of using the electromyography method in nutrition science].
Investigation of the electrical activity of smooth muscle in disorders of motor function of the gastrointestinal tract is a modern method which let to assess not only the function, but also violations of related systems. Particularly, the level of electromotor activity can be measure of the rate of intestinal transit and its effect on the absorption of nutrients from different regions.. Non invasivity of the method allows its repeated use in difficult diagnostic cases.


Equity of renal replacement therapy utilization: a prospective population-based study.
This 1-year prospective survey assessed the incidence and characteristics of all patients starting renal replacement therapy (RRT) for end-stage renal disease in Scotland, and whether there is equity of utilization of RRT in terms of age, domicile and social circumstance. In the year studied, 104 patients per million population (533 patients) started RRT (390 per million population aged 65-75). In 23.5% the cause of ESRD could not be determined. Diabetes was the single most frequently identified cause (16%). The requirement for RRT rose with age, but over the country as a whole, patients aged over 75 years were under-represented. The majority of health boards provided RRT at a rate within 20% of the national rate. There was no difference in the median age at starting RRT between health boards. The spectrum of social deprivation of patients starting RRT was the same as that of the general population. There was no evidence that social deprivation influences acceptance on to the RRT program, although the relationship between ESRD and deprivation is complex. The utilization of RRT exceeded the minimum rate recommended by the Renal Association, although there was fluctuation between health board areas. The national requirement for resources to provide RRT is likely to rise further to care for an increasingly elderly population.


Zonisamide discontinuation due to psychiatric and cognitive adverse events: a case-control study.
Zonisamide (ZNS) is an antiepileptic drug (AED) that has been associated with psychiatric adverse events (PAE) and cognitive adverse events (CAE); controlled studies evaluating these adverse events are limited. Our objectives were to 1) determine the incidence of PAE and CAE leading to the discontinuation of ZNS and 2) identify risk factors for PAE and CAE associated with the discontinuation of ZNS. All patients exposed to ZNS at MINCEP Epilepsy Care between March 2000 and September 2008 were identified. Reasons for discontinuing ZNS were documented. Separate case-control studies were performed to identify risk factors associated with the discontinuation of ZNS due to PAE or CAE via multivariate binary logistic regression. A total of 544 patients were exposed to ZNS during the study period. PAE and CAE were the most frequently identified reasons for terminating ZNS therapy. The incidence of PAE severe enough to be associated with the discontinuation of ZNS was 6.9%; the incidence of CAE was 5.8%. Factors associated with termination of ZNS therapy due to PAE were past psychiatric history (p = 0.005), symptomatic generalized epilepsy (p = 0.027), and lower maximum ZNS serum concentration (mean = 17.9 mg/L vs 34.7 mg/L, p < 0.001). Independent variables associated with discontinuing ZNS due to CAE were greater number of concomitant AEDs (p = 0.011) and lower maximum ZNS serum concentration (mean = 16.6 mg/L vs 30.6 mg/L, p = 0.002). We have identified clinically relevant risk factors associated with the discontinuation of ZNS. Our findings support the concept that selected patients are relatively more vulnerable to CNS adverse events when exposed to ZNS.


Terminally differentiated effector memory T cells associate with cognitive and AD-related biomarkers in an aging-based community cohort.
The immune response changes during aging and the progression of Alzheimer's disease (AD) and related dementia (ADRD). Terminally differentiated effector memory T cells (called T EMRA ) are important during aging and AD due to their cytotoxic phenotype and association with cognitive decline. However, it is not clear if the changes seen in T EMRAs are specific to AD-related cognitive decline specifically or are more generally correlated with cognitive decline. This study aimed to examine whether T EMRAs are associated with cognition and plasma biomarkers of AD, neurodegeneration, and neuroinflammation in a community-based cohort of older adults. Study participants from a University of Kentucky Alzheimer's Disease Research Center (UK-ADRC) community-based cohort of aging and dementia were used to test our hypothesis. There were 84 participants, 44 women and 40 men. Participants underwent physical examination, neurological examination, medical history, cognitive testing, and blood collection to determine plasma biomarker levels (Aβ42/Aβ40 ratio, total tau, Neurofilament Light chain (Nf-L), Glial Fibrillary Acidic Protein (GFAP)) and to isolate peripheral blood mononuclear cells (PBMCs). Flow cytometry was used to analyze PBMCs from study participants for effector and memory T cell populations, including CD4 + and CD8 + central memory T cells (T CM ), Naïve T cells, effector memory T cells (T EM ), and effector memory CD45RA + T cells (T EMRA ) immune cell markers. CD8 + T EMRAs were positively correlated with Nf-L and GFAP. We found no significant difference in CD8 + T EMRAs based on cognitive scores and no associations between CD8 + T EMRAs and AD-related biomarkers. CD4 + T EMRAs were associated with cognitive impairment on the MMSE. Gender was not associated with T EMRAs , but it did show an association with other T cell populations. These findings suggest that the accumulation of CD8 + T EMRAs may be a response to neuronal injury (Nf-L) and neuroinflammation (GFAP) during aging or the progression of AD and ADRD. As our findings in a community-based cohort were not clinically- defined AD participants but included all ADRDs, this suggests that T EMRAs may be associated with changes in systemic immune T cell subsets associated with the onset of pathology.


Unconjugated bilirubin exposure impairs hippocampal long-term synaptic plasticity.
Jaundice is one of the most common problems encountered in newborn infants, due to immaturity of hepatic conjugation and transport processes for bilirubin. Although the majority of neonatal jaundice is benign, some neonates with severe hyperbilirubinemia develop bilirubin encephalopathy or kernicterus. Accumulation of unconjugated bilirubin (UCB) in selected brain regions may result in temporary or permanent impairments of auditory, motor, or cognitive function; however, the molecular mechanisms by which UCB elicits such neurotoxicity are still poorly understood. The present study is undertaken to investigate whether prolonged exposure of rat organotypic hippocampal slice cultures to UCB alters the induction of long-term synaptic plasticity. Using electrophysiological recording techniques, we find that exposure of hippocampal slice cultures to clinically relevant concentrations of UCB for 24 or 48 h results in an impairment of CA1 long-term potentiation (LTP) and long-term depression (LTD) induction in a time- and concentration-dependent manner. Hippocampal slice cultures stimulated with UCB show no changes in the secretion profiles of the pro-inflammatory cytokines, interleukin-1beta and tumor necrosis factor-alpha, or the propidium ioide uptake. UCB treatment produced a significant decrease in the levels of NR1, NR2A and NR2B subunits of N-methyl-D-aspartate (NMDA) receptors through a calpain-mediated proteolytic cleavage mechanism. Pretreatment of the hippocampal slice cultures with NMDA receptor antagonist or calpain inhibitors effectively prevented the UCB-induced impairment of LTP and LTD. Our results indicate that the proteolytic cleavage of NMDA receptor subunits by calpain may play a critical role in mediating the UCB-induced impairment of long-term synaptic plasticity in the hippocampus. These observations provide new insights into the molecular mechanisms underlying UCB-induced impairment of hippocampal synaptic plasticity which, in turn, might provide opportunities for the development of novel therapeutic strategies that targets these pathways for treatment.


Phytosterols promote liver injury and Kupffer cell activation in parenteral nutrition-associated liver disease.
Parenteral nutrition-associated liver disease (PNALD) is a serious complication of PN in infants who do not tolerate enteral feedings, especially those with acquired or congenital intestinal diseases. Yet, the mechanisms underlying PNALD are poorly understood. It has been suggested that a component of soy oil (SO) lipid emulsions in PN solutions, such as plant sterols (phytosterols), may be responsible for PNALD, and that use of fish oil (FO)-based lipid emulsions may be protective. We used a mouse model of PNALD combining PN infusion with intestinal injury to demonstrate that SO-based PN solution causes liver damage and hepatic macrophage activation and that PN solutions that are FO-based or devoid of all lipids prevent these processes. We have furthermore demonstrated that a factor in the SO lipid emulsions, stigmasterol, promotes cholestasis, liver injury, and liver macrophage activation in this model and that this effect may be mediated through suppression of canalicular bile transporter expression (Abcb11/BSEP, Abcc2/MRP2) via antagonism of the nuclear receptors Fxr and Lxr, and failure of up-regulation of the hepatic sterol exporters (Abcg5/g8/ABCG5/8). This study provides experimental evidence that plant sterols in lipid emulsions are a major factor responsible for PNALD and that the absence or reduction of plant sterols is one of the mechanisms for hepatic protection in infants receiving FO-based PN or lipid minimization PN treatment. Modification of lipid constituents in PN solutions is thus a promising strategy to reduce incidence and severity of PNALD.


Juvenile hyaline fibromatosis: an unusual clinical presentation.
Juvenile hyaline fibromatosis is a recessive autosomal hereditary disorder characterized by abnormal growth of hyalinized fibrous tissue. Its clinical presentation is marked by tumors of the skin, bone lesions, joint contractures, and gingival hyperplasia. We report a localized form of juvenile hyaline fibromatosis, a rare disease with several cases reported in the worldwide literature. A 23-year-old man presented with multiple tumors, joint contractures, and osteolytic bone lesions, but without gingival hyperplasia in one year of follow-up. Although, the onset of this condition is commonly in early childhood with progression, his unusual clinical presentation began at eight years of age with late progression in adolescence.


Impairment but not abolishment of express saccades after unilateral or bilateral inactivation of the frontal eye fields.
Express saccades are a manifestation of a visual grasp reflex triggered when visual information arrives in the intermediate layers of the superior colliculus (SCi), which in turn orchestrates the lower level brainstem saccade generator to evoke a saccade with a very short latency (~100 ms or less). A prominent theory regarding express saccades generation is that they are facilitated by preparatory signals, presumably from cortical areas, which prime the SCi before the arrival of visual information. Here, we test this theory by reversibly inactivating a key cortical input to the SCi, the frontal eye fields (FEF), while monkeys perform an oculomotor task that promotes express saccades. Across three tasks with a different combination of potential target locations and unilateral or bilateral FEF inactivation, we found a spared ability for monkeys to generate express saccades, despite decreases in express saccade frequency during FEF inactivation. This result is consistent with the FEF having a facilitatory but not critical role in express saccade generation, likely because other cortical areas compensate for the loss of preparatory input to the SCi. However, we also found decreases in the accuracy and peak velocity of express saccades generated during FEF inactivation, which argues for an influence of the FEF on the saccadic burst generator even during express saccades. Overall, our results shed further light on the role of the FEF in the shortest-latency visually-guided eye movements.NEW & NOTEWORTHY Express saccades are the shortest-latency saccade. The frontal eye fields (FEF) are thought to promote express saccades by presetting the superior colliculus. Here, by reversibly inactivating the FEF either unilaterally or bilaterally via cortical cooling, we support this by showing that the FEF plays a facilitative but not critical role in express saccade generation. We also found that FEF inactivation lowered express saccade peak velocity, emphasizing a contribution of the FEF to express saccade kinematics.


Efficient removal of bilirubin from human serum by monosize dye affinity beads.
Cibacron Blue F3GA (CB) was covalently attached onto poly(glycidyl methacrylate) (PGMA) monosize beads for removal of bilirubin from hyperbilirubinemia human serum. PGMA beads were produced by dispersion polymerization (1.6 μm in diameter). CB loading was 1.73 mmol/g. Bilirubin adsorption experiments were performed by stirred-batch adsorption. The non-specific adsorption of bilirubin was low (0.4 mg/g polymer). CB attachment onto the PGMA beads significantly increased the bilirubin adsorption (241.5 mg/g) from aqueous solutions. The maximum bilirubin adsorption was observed at pH 6.0. With an increase of the aqueous phase concentration of sodium chloride, the adsorption amount of bilirubin decreased drastically. The equilibrium adsorption of bilirubin significantly increased with increasing temperature. Much higher adsorption values up to 332 mg bilirubin/g were achieved in the case of the PGMA/CB beads from human plasma.


The Effects of Dienogest on Macrophage and Natural Killer Cells in Adenomyosis: A Randomized Controlled Study.
Progestin has been used for symptomatic treatment of adenomyosis, although its effect on the immune system has not been studied. The aim of this study was to investigate the changes of macrophage and natural killer (NK) cell infiltration in tissues obtained from women with adenomyosis who did or did not receive oral progestin dienogest. In this randomized controlled clinical trial study, 24 patients with adenomyosis who required hysterectomy were enrolled. Twelve patients received dienogest 28-35 days before surgery, and the other 12 patients were not treated with any hormones. The endometrial and myometrial tissue samples were immediately collected after hysterectomy, and immunohistochemistry for a macrophage marker (CD68) and a NK cells marker (CD57) was performed. The number of CD57 cells was significantly increased in endometrial glands of the treated group compared to the untreated group (P=0.005) but not in stroma in the endometrium of the treated patients (P=0.416). The difference in the number of CD68 cells was not statistically significant between treated and untreated groups in the endometrial glands (P=0.055) or stromal tissues (P=0.506). Administration of oral progestin dienogest to patients with adenomyosis increased the number of uterine infiltrating NK cells in glandular structure of eutopic endometrium. The differential effects of progestin on NK cells depended on the site of immune cell infiltration. The effects of oral progestin on uterine NK cells in adenomyosis have the potentials to be beneficial to pregnancies occurring following discontinuation of treatment in terms of embryo implantation and fetal protection (Registration number: TCTR20150921001).


Dimensions of temperament modulate cue-controlled behavior: a study on Pavlovian to instrumental transfer in horses (Equus caballus).
Pavlovian to instrumental transfer (PIT) is a central factor in how cues influence animal behavior. PIT refers to the capacity of a Pavlovian cue that predicts a reward to elicit or increase a response intended to obtain the same reward. In the present study, using an equine model, we assessed whether PIT occurs in hoofed domestic animals and whether its efficacy can be modulated by temperamental dimensions. To study PIT, horses were submitted to Pavlovian conditioning whereby an auditory-visual stimulus was repeatedly followed by food delivery. Then, horses were submitted to instrumental conditioning during which they learned to touch with their noses an object signaled by the experimenter in order to obtain the same reward. During the PIT test, the Pavlovian conditioned stimulus was presented to the animal in the absence of reward. At the end of the experiment, a battery of behavioral tests was performed on all animals to assess five temperamental dimensions and investigate their relationships with instrumental performance. The results indicate that PIT can be observed in horses and that its efficacy is greatly modulated by individual temperament. Indeed, individuals with a specific pattern of temperamental dimensions (i.e., higher levels of gregariousness, fearfulness, and sensory sensitivity) exhibited the strongest PIT. The demonstration of the existence of PIT in domesticated animals (i.e., horses) is important for the optimization of its use by humans and the improvement of training methods. Moreover, because PIT may be implicated in psychological phenomena, including addictive behaviors, the observation of relationships between specific temperamental dimensions and PIT efficacy may aid in identifying predisposing temperamental attributes.


Response of biomarkers in amphibian larvae to in situ exposures in a fruit-producing region in North Patagonia, Argentina.
The authors evaluated biomarker responses in caged larvae of the amphibian Rhinella arenarum in water channels during fruit production season and compared them with those elicited by a transient exposure to azinphos methyl (AzM) (0.02-2 mg/L; 4 h), the main pesticide applied in the Alto Valle region, Patagonia, Argentina, taking into account the maximum environmental concentration detected in superficial water (22.5 µg/L). The traditional biomarkers of organophosphate exposure, acetylcholinesterase (AChE) and carboxylesterase, were inhibited in tadpoles after one week of exposure in channels potentially receiving pesticide drift, whereas the antioxidant glutathione (GSH) and the detoxifying activity of GSH S-transferase (GST) were induced. In a two-week monitoring study, AChE activity was induced in larvae exposed at the agricultural site, and carboxylesterase showed an inhibition followed by return to control values, suggesting an exposure-recovery episode. Antioxidant glutathione levels were first depleted and then surpassed control levels, whereas GST activity was continuously induced. These responses were mimicked in the laboratory by 2 mg/L AzM-pulse exposure, which notably exceeds the expected environmental concentrations. The results draw attention to the complexity of responses after pesticide exposure, strongly depending on exposure time-concentration and recovery periods, among other possible factors, and support the necessity of the integrated use of biomarkers to assess exposure episodes in agricultural areas.


Head Start Immunity: Characterizing the Early Protection of C Strain Vaccine Against Subsequent Classical Swine Fever Virus Infection.
Classical Swine Fever Virus (CSFV) is an ongoing threat to the pig industry due to the high transmission and mortality rates associated with infection. Live attenuated vaccines such as the CSFV C strain vaccine are capable of protecting against infection within 5 days of vaccination, but the molecular mechanisms through which this early protection is mediated have yet to be established. In this study, we compared the response of pigs vaccinated with the C strain to non-vaccinated pigs both challenged with a pathogenic strain of CSFV. Analysis of transcriptomic data from the tonsils of these animals during the early stages after vaccination and challenge reveals a set of regulated genes that appear throughout the analysis. Many of these are linked to the ISG15 antiviral pathway suggesting it may play a role in the rapid and early protection conferred by C strain vaccination.


Abacus: a computational tool for extracting and pre-processing spectral count data for label-free quantitative proteomic analysis.
We describe Abacus, a computational tool for extracting spectral counts from MS/MS data sets. The program aggregates data from multiple experiments, adjusts spectral counts to accurately account for peptides shared across multiple proteins, and performs common normalization steps. It can also output the spectral count data at the gene level, thus simplifying the integration and comparison between gene and protein expression data. Abacus is compatible with the widely used Trans-Proteomic Pipeline suite of tools and comes with a graphical user interface making it easy to interact with the program. The main aim of Abacus is to streamline the analysis of spectral count data by providing an automated, easy to use solution for extracting this information from proteomic data sets for subsequent, more sophisticated statistical analysis.


Age-related effects in the marking of old objects in visual search.
Two studies were performed to examine potential age-related differences in visual marking. Visual marking is a limited capacity process that enhances visual search through the inhibition of old objects. In Experiment 1, young and old adults performed three different search conditions: a full-element baseline, a half-element baseline, and a gap condition in which they searched through displays similar to the full-element baseline condition but with half of the letters presented before and the other half of the letters and the target presented after a 1000-ms gap. Both old and young adults displayed search slopes in the gap condition that were equivalent to slopes obtained in the half-element condition, suggesting that they were able to successfully inhibit the old letters. In Experiment 2, old and young adults also performed in three different visual search conditions but in this case with targets defined either by a form difference or by a conjunction of form and color. Both old and young adults showed a reduced slope in the gap as compared to the conjunction condition, suggesting inhibition of the old objects. The data are discussed in terms of age-related differences in the top-down control of attention in visual search.


Graphene-graphene oxide floating gate transistor memory.
A novel transparent, flexible, graphene channel floating-gate transistor memory (FGTM) device is fabricated using a graphene oxide (GO) charge trapping layer on a plastic substrate. The GO layer, which bears ammonium groups (NH3+), is prepared at the interface between the crosslinked PVP (cPVP) tunneling dielectric and the Al2 O3 blocking dielectric layers. Important design rules are proposed for a high-performance graphene memory device: (i) precise doping of the graphene channel, and (ii) chemical functionalization of the GO charge trapping layer. How to control memory characteristics by graphene doping is systematically explained, and the optimal conditions for the best performance of the memory devices are found. Note that precise control over the doping of the graphene channel maximizes the conductance difference at a zero gate voltage, which reduces the device power consumption. The proposed optimization via graphene doping can be applied to any graphene channel transistor-type memory device. Additionally, the positively charged GO (GO-NH3+) interacts electrostatically with hydroxyl groups of both UV-treated Al2 O3 and PVP layers, which enhances the interfacial adhesion, and thus the mechanical stability of the device during bending. The resulting graphene-graphene oxide FGTMs exhibit excellent memory characteristics, including a large memory window (11.7 V), fast switching speed (1 μs), cyclic endurance (200 cycles), stable retention (10(5) s), and good mechanical stability (1000 cycles).


Silencing the jasmonate cascade: induced plant defenses and insect populations.
We transformed the native tobacco, Nicotiana attenuata, to silence its lipoxygenase, hydroperoxide lyase, and allene oxide synthase genes in order to inhibit oxylipin signaling, known to mediate the plant's direct and indirect defenses. When planted into native habitats, lipoxygenase-deficient plants were more vulnerable to N. attenuata's adapted herbivores but also attracted novel herbivore species, which fed and reproduced successfully. In addition to highlighting the value of genetically silencing plants to study ecological interactions in nature, these results show that lipoxygenase-dependent signaling determines host selection for opportunistic herbivores and that induced defenses influence herbivore community composition.


A new algorithm for image edge extraction using a statistical classifier approach.
This correspondence describes a new algorithm for extracting edges from natural images. Starting from a simple image model, the algorithm poses the problem of edge extraction as a statistical classifier problem. The algorithm is capable of extracting and detecting edges from images even in the presence of noise. A step by step mathematical derivation of the algorithm reveals the flexibility of the algorithm with pertinent parameters that can be varied for the specific need of the user. Finally, the proposed edge operator is compared to the well-known Marr-Hildreth's edge operator.


Complete Genome Sequence of Lactococcus lactis subsp. lactis A12, a Strain Isolated from Wheat Sourdough.
We report here the complete genome sequence of Lactococcus lactis subsp. lactis strain A12, a strain isolated from sourdough. The circular chromosome and the four plasmids reveal genes involved in carbohydrate metabolism that are potentially required for the persistence of this strain in such a complex ecosystem.


[Histophysiology of the zona fasciculata of the adrenal cortex of albino rats].
Three phases have been detected in the fascicular zone of the white rat adrenal cortex. During the first phase (normoemia) blood stream is rather moderate, lipids accumulate in adrenocorticocytes. This results in increase of their volume and in decrease of nuclear-cytoplasmic relations. The second phase (functional hyperemia) is characterized with an elevated blood stream and plethora, maximal parameters of nuclear volume and nuclear-cytoplasmic relations, decreasing content of lipids and volume of adrenocorticocytes. During the third phase (functional hypoemia) parameters characterising intensity of blood stream, nuclear volume and nuclear-cytoplasmic relations decrease.


Genetic variation in coding regions between and within commonly used inbred rat strains.
Single nucleotide polymorphisms (SNPs) are the most common genetic variation in mammalian populations. Their significance is illustrated by their potential contribution to common disease but also by their potential for use in genetic association and mapping experiments. We have examined the genetic variation between commonly used inbred rat strains by using an efficient SNP discovery and typing assay based on enzyme-based (CEL I) heteroduplex cleavage. Screening of a panel of 96 different rat (sub-)strains for 100 genomic loci in 55 genes, whose human homologs are implicated in clinically relevant diseases like neurological disorder, cancer, schizophrenia, and obesity, resulted in the identification of 103 novel polymorphisms. As all strains are simultaneously genotyped in this setup, this allowed us to make an estimate of the genetic variation between and within commonly used rat inbred strains. Interestingly, we observed substantial genetic variation between colonies of the same inbred strain, maintained at different locations. Furthermore, we identified 17 non-synonymous SNPs that may have an effect on protein function and contribute to phenotypic differences between different laboratory strains.


[Mitochondrial defects].
Mitochondrial myopathies are characterized by structural and functional abnormalities of the mitochondria. The diseases are best classified according to the underlying biochemical defect. In this article the variability of clinical expression and mitochondrial abnormalities is illustrated. An exact diagnosis is a prerequisite for a rational therapy and genetic counseling. In exceptional cases antenatal diagnosis should be possible.


Contrast enhancement with uranyl acetate allows quantitative analysis of the articular cartilage by microCT: Application to mandibular condyles in the BTX rat model of disuse.
Microcomputed tomography (microCT) is well adapted to quantitative analysis of calcified tissues but soft tissues (such as cartilage) are radiolucent and need a contrast enhancement procedure for microCT. We developed a "staining" method allowing microCT imaging of articular cartilage using uranyl acetate (UA). The method was used to see whether adult rats with a botulinum toxin (BTX) injection in masticatory muscles present a change at the condylar cartilage of the mandible in association with a localized trabecular bone loss. Human femoral head samples were used to develop the enhanced contrast method using UA or lanthanides (recently proposed as a substitute for UA). The method was then applied to the condylar cartilage of rat mandibles. Mature male rats (n=11) were randomized into 2 groups: control (CTRL; n=4) and BTX group (n=7). Rats of the BTX group received a single injection of BTX into the right M. Masseter and M. Temporalis. Rats of the CTRL group were similarly injected with saline. Rats were sacrificed 4 weeks after injection. Condyles were harvested, fixed in formalin and immersed in UA. MicroCT was performed for bone and cartilage measurements. After UA impregnation, articular cartilage of human femoral head samples was clearly seen on its full thickness whereas lanthanides produced a much less pronounced contrast, with a faint labeling at the upper layer. In BTX rats, microCT analysis showed a significant bone loss at the right condyles. After UA, the whole thickness of articular cartilage was clearly evidenced. Cartilage thickness measurement showed no difference when comparing the right with the left sides of the BTX group nor between the two sides of the CTRL group. Contrast enhancement with UA is a simple technique allowing quantitative analysis of cartilage by microCT.290 words.


Low-dimensional functional networks of cage-like B40 with effective transition-metal intercalations.
As the first all-boron fullerene observed in experiments, cage-like borospherene B40 has attracted considerable attention in recent years. However, B40 has been proved to be chemically reactive and tends to coalesce with one another via the formation of covalent bonds. We explore herein the possibility of low-dimensional functional networks of B40 with effective transition-metal intercalations. We find that the four equivalent B7 heptagons on the waist of each B40 can serve as effective ligands to coordinate various transition metal centers in exohedral motifs. The intercalated metal atoms entail these networks with a variety of intriguing properties. The two-dimensional (2D) Cr2B40 network is a ferromagnetic metal while the 2D Zn2B40 network becomes semiconducting. In contrast, other 2D M2B40 (M = Sc, Ti, V, Mn, Fe, Co, Ni and Cu) networks and 1D CrB40 belong to nonmagnetic metals. The 3D Cr3B40 network is a magnetic metal. This work presents the viable possibility of assembling Mn&B40 metalloborospherenes into stable functional nanomaterials via effective transition-metal intercalations with potential applications in electronic and spintronic devices.


The Impact of Self-Criticism and Self-Reassurance on Weight-Related Affect and Well-Being in Participants of a Commercial Weight Management Programme.
Certain psychological and emotional factors can undermine attempts at weight management. Previously we have found that shame and self-criticism were significantly associated with disinhibition and perceived hunger in 2,236 participants of a weight management programme. This effect was fully mediated through weight-related negative affect. The present study examined the impact of self-criticism and self-reassurance on well-being and whether it was mediated by weight-related affect in the same population. Participants completed an online survey of measures of self-criticism and self-reassurance, and negative and positive affect associated with weight and well-being. Path analysis suggested that self-criticism was significantly associated with decreased well-being, both directly and indirectly, mediated by increased negative and decreased positive weight-related affect. Self-reassurance had a stronger association with increased well-being by predicting lower negative and increased positive weight-related affect. All effects were significant at p < 0.001. Self-criticism and self-reassurance were related to well-being in participants attempting to manage their weight, both directly and through their impact on weight-related affect. The positive association between self-reassurance and well-being was stronger than the negative association between self-criticism and well-being. Supporting the development of self-reassuring competencies in weight management programmes may improve weight-related affect and well-being.


Anoxia hormesis following overwintering diapause boosts bee survivorship and adult performance.
Insect pollination is a crucial component of our ecosystems and biodiversity, but our reliance on this ecosystem service has much broader implications. We depend on these pollination services to produce materials and food. But insect pollinators, especially bees, are in strong decline due to a plethora of factors, least of which are environmental abiotic stressors like climate change. The alfalfa leafcutting bee, Megachile rotundata, is the world's most managed solitary bee and is particularly vulnerable to changes in temperature. This species spends up to ten months overwintering while being exposed to the lowest temperatures of winters and the hottest temperatures of late summer. This results in usage of energy reserves prematurely and asynchronous spring emergence with their food resource. To understand the stress response of these bees and potentially boost their performance, we applied a hormetic framework where bees were exposure to different doses of anoxia (the absence of oxygen) to trigger hormesis; a low-dose stimulatory response known to lower damage and improve performance. We used hormesis on immature bees as a post-winter treatment with the goal of improving springtime performance in adults. One hour of anoxia had no negative effect on adult springtime emergence and bees were quick to recover. These bees were more active than untreated bees, as resistant to starvation, and as long-lived. Higher exposure to anoxia (3 h) was found to be mildly hormetic and 6-h exposures were detrimental. Anoxia hormesis did not represent a significant cost on the energy reserve of overwintering bees but we found that the age at which anoxia is applied will affect the effectiveness of treatment. Our data suggest that anoxia hormesis is a viable intervention to improve springtime performance in overwintering bees.


Application of the direct peptide reactivity assay (DPRA) to inorganic compounds: a case study of platinum species.
The in chemico Direct Peptide Reactivity Assay (DPRA) was developed as a non-animal, relatively high throughput, screening tool for skin sensitization potential. Although the Adverse Outcome Pathway (AOP) for respiratory sensitization remains to be fully elucidated, it is recognized that the molecular initiation event for both skin and respiratory sensitization to low molecular weight chemicals involves haptenation with proteins. The DPRA examines the reactivity of a test compound to two model peptides (containing either cysteine or lysine) and consequently is able to screen for both skin and respiratory sensitization potential. The DPRA was primarily developed for and validated with organic compounds and assessment of the applicability of the assay to metal compounds has received only limited attention. This paper reports the successful application of the DPRA to a series of platinum compounds, including hexachloroplatinate and tetrachloroplatinate salts, which are some of the most potent chemical respiratory sensitizers known. Eleven platinum compounds were evaluated using the DPRA protocol as detailed by Lalko et al., with only minor modification. Two palladium compounds with structures similar to that of the platinum species studied and cobalt chloride were additionally tested for comparison. The hexachloroplatinate and tetrachloroplatinate salts showed exceptionally high reactivity with the cysteine peptide (EC15 values of 1.4 and 14 μM, respectively). However, for platinum compounds (e.g. hydrogen hexahydroxyplatinate and tetraammineplatinum) where clinical and epidemiological evidence indicates limited sensitization potential, the cysteine DPRA showed only minor or no reactivity (EC15 values of 24 600 and >30 000 μM, respectively). The outcomes of the lysine peptide assays were less robust and where EC15 was measurable, values were substantially higher than the corresponding results from the cysteine assay. This work supports the value of in chemico peptide reactivity as a metric for assessment of platinum sensitization potential and therefore in screening of new platinum compounds for low or absent sensitization potential. Additional studies are required to determine whether the DPRA may be successfully applied to other metals. We provide details on method modifications and precautions important to the success of the DPRA in the assessment of metal reactivity.