A quality improvement project to increase palliative care team involvement in pediatric oncology patients.
Pediatric palliative care (PPC) for oncology patients improves quality of life and the likelihood of goal-concordant care. However, barriers to involvement exist. We aimed to increase days between PPC consult and death for patients with refractory cancer from a baseline median of 13.5 days to ≥30 days between March 2019 and March 2020. Outcome measure was days from PPC consult to death; process measure was days from diagnosis to PPC consult. The project team surveyed oncologists to identify barriers. Plan-do-study-act cycles included establishing target diagnoses, offering education, standardizing documentation, and sending reminders. The 24-month baseline period included 30 patients who died and 25 newly diagnosed patients. The yearlong intervention period included six patients who died and 16 newly diagnosed patients. Interventions improved outcome and process measures. Targeted patients receiving PPC ≥30 days prior to death increased from 43% to 100%; median days from consult to death increased from 13.5 to 159.5. Targeted patients receiving PPC within 30 days of diagnosis increased from 28% to 63%; median days from diagnosis to consult decreased from 221.5 to 14. Of those without PPC consult ≤ 30 days after diagnosis, 17% had template documentation of the rationale. Interventions utilized met the global aim, outcome, and process measures. Use of QI methodology empowered providers to involve PPC. Poor template use was a barrier to identifying further drivers. Future directions for this project relate to expanding the target list, creating long-term sustainability, formalizing standards, and surveying patients and families.


Kinetically controlled synthesis of monoglyceryl esters from chiral and prochiral acids methyl esters catalyzed by immobilized Rhizomucor miehei lipase.
Partial acylation of only one primary hydroxyl group of glycerol generates a chiral center at position 2. Rhizomucor miehei lipase (RML) catalyzes the kinetically controlled transesterification of different aromatic carboxylic acids methyl esters with glycerol. High synthetic yields of glyceryl esters (around 70-80%) were obtained even in the presence of significant concentrations of water (from 5% to 20%). After a long incubation of the reaction mixture in the presence of the biocatalyst only pure free acid was obtained. Other lipases (from Geobacillus thermocatenulatus and from Thermomyces lanuginose) also catalyzed similar kinetically controlled transesterifications although less efficiently. RML immobilized on Sepharose-Q showed a high activity and specificity, compared to the immobilization by other techniques, only producing monoglyceryl esters with all substrates. In particular, monoglyceryl-phenylmalonate product was synthesized in 82% overall yield and >99% diastereomeric excess at pH 7.0 and 37°C and 90% glycerol.


Differential Long-Term Effects of First- and Second-Generation DES in Patients With Bifurcation Lesions Undergoing PCI.
There is a paucity of data regarding the long-term clinical outcomes of first- versus second-generation drug-eluting stent (DES), especially when used to treat complex lesions such as bifurcation lesions. The current study compares the efficacy and safety of first- versus second-generation DES at the 5-year follow-up in patients who underwent bifurcation percutaneous coronary intervention (PCI). A total of 5,498 patients with a bifurcation lesion who underwent PCI were pooled at a single patient level from COBIS (Coronary Bifurcation Stenting) registries II and III. Five-year target lesion failure (TLF) (the composite of cardiac death, myocardial infarction [MI], and target lesion revascularization [TLR]) and cardiac death or MI were compared between the use of first-generation DES (n = 2,436) and second-generation DES (n = 3,062) during PCI. Propensity score matching was performed to reduce selection bias. After a 1:1 propensity score matching procedure was conducted, the cohort consisted of 1,702 matched pairs. Patients treated with second-generation DES had a significantly lower risk of TLF at 5 years than those treated with first-generation DES in both overall and propensity-matched populations (matched hazard ratio [HRmatched]: 0.576; 95% confidence interval [CI]: 0.456 to 0.727; p <0.001). There were no significant differences in risk of a composite of cardiac death or MI between the 2 groups (HRmatched: 0.782; 95% CI: 0.539 to 1.133, P = 0.193). However, among patients who required a 2-stent technique, use of the second-generation DES reduced cardiac death or MI (HRmatched:0.422; 95% CI: 0.209 to 0.851, P = 0.016). On the other hand, among patients who required a one-stent technique, the risk of a composite of cardiac death or MI was similar between the 2 groups (HRmatched: 1.046; 95% CI: 0.664 to 1.650, P = 0.845). There was a significant interaction between stent generation and treatment strategy for cardiac death or MI (interaction P = 0.029). In patients treated with PCI for a bifurcation lesion, the use of second-generation DES was associated with a significantly reduced risk of 5-year TLF than the use of first-generation DES. (Korean Coronary Bifurcation Stenting Registry II [NCT01642992]; COBIS II) (Korean Coronary Bifurcation Stenting Registry III [NCT03068494] COBIS III).


Binding and mobility of mercury in soils contaminated by emissions from chlor-alkali plants.
Chlor-alkali plants are known to be an important source of Hg emissions to the atmosphere and related contamination of soils in their vicinity. In the present study, the results of Hg speciation and mobility of Hg in soils affected by Hg emissions from three chlor-alkali plants are compared. Solid phase mercury speciation analyses was carried out using a mercury-thermo-desorption technique with the aim of distinguishing elemental Hg [Hg(0)] from Hg(II)-binding forms. Mercury species in soil leachates were distinguished using an operationally defined method, which is based on the reactivity of soluble Hg compounds. Results show that the Hg(0) emitted from the plants could not be detected in any of the investigated soils. This indicates quantitative re-emission or oxidation of this Hg species in the atmosphere or soils. In most soils Hg was predominately bound to organic matter. Only in sandy soils deficient in organic matter was Hg, to a larger extent, sorbed onto mineral soil components. Leachable Hg in most soils occurred as non-reactive, soluble organic Hg complexes such as fulvic acid-bound Hg, and reach their highest values (90 microg kg(-1)) in soils rich in organic matter. Concentrations of reactive, soluble Hg compounds were highest in sandy soils where the content of organic matter was low. Leachability of Hg was found to be inhibited in soils with a high content of clayey soil components. The distribution of Hg in soil profiles suggests that migration of Hg to deeper soil layers (approx. 20 cm) is most effective if Hg is bound to soluble organic complexes, whereas reactive Hg or weak Hg complexes are effectively retained in the uppermost soil layer (5 cm) through sorption on mineral surfaces.


Possible anti-depressant effect of efavirenz and pro-depressive-like effect of voriconazole in specified doses in various experimental models of depression in mice.
Reports from experimental and clinical studies have indicated the possible relation between cholesterol and depression. Efavirenz (EFV) and Voriconazole (VRC) have been reported to affect cholesterol-24S-hydroxylase enzyme. The objective of the present study was to evaluate the effect of EFV and VRC in experimental models of depression in mice. There was a measurement of immobility time in forced swim test and tail suspension test in which mice were previously subjected to the treatment of EFV (0.09mg/kg, orally (po)) and VRC (75mg/kg, intraperitoneally (ip)) separately for 14days. Sucrose intake was measured during stress schedule of 21days in chronic mild stress model in which mice were subjected to above mentioned drug treatment for last 14days. There was an estimation of serum total cholesterol and brain serotonin levels on day 21. The results indicated that mice treated with EFV showed a significant decrease in the immobility time and increase in sucrose intake with decrease in serum total cholesterol. Mice treated with VRC showed a significant increase in the immobility time and decrease in the sucrose intake with increase in serum total cholesterol. There was a significant increase and decrease in brain serotonin levels in mice treated with EFV and VRC respectively. The results of the present study indicates the possible anti-depressant effect of EFV and pro-depressive-like effect of VRC in specified doses in mice, raising the possibility that stimulation but not inhibition of cholesterol-24S-hydroxylase may be important in the treatment of depression.


Usefulness of the FDG PET/CT in the management of cystic echinococcosis: A pilot study.
Cystic echinococcosis is a zoonotic disease caused by the cestode Echinococcus granulosus. The activity of the cysts is assessed through the WHO-IWGE standardized classification based on ultrasound features. However, viability of the cysts is not always concordant with the activity assessed by ultrasound. The aim of the present study is to describe the metabolic activity of cysts in patients with cystic echinococcosis through FDG-PET. Prospective observational study where adult patients diagnosed of cystic echinococcosis were offered to undergo FDG PET/CT before treatment onset. Demographic, clinical, radiological, and histopathological information was collected from all patients. Sixteen patients were included, 50% were male, and age ranged from 18 to 85 years. Most of the patients had liver involvement, and all patients had CE3, CE4 or CE5 stage of the WHO-IWGE classification. Only one patient (CE5) had an increased 18F-FDG uptake of the cyst in the FDG PET/CT. From the 5 patients who underwent surgical treatment, only one showed signs of viability of the cyst: a patient with CE5 with no increased 18F-FDG uptake of the cyst. In our pilot study, we did not find the correlation between the FDG PET/CT imaging and the cystic echinococcosis cyst bioactivity.


Physicochemical and molecular transformation of novel functional peptides from Baijiu.
A novel strategy for screening and identifying peptides present in Baijiu was developed based on magnetic solid-phase extraction with magnetic S-doped graphene (M-G-S) as adsorbent combined with ultrahigh-performance liquid chromatography with high resolution tandem mass spectrometry. In total, 28 peptides consisting of amino acids from 3 to 9 were preliminarily identified, and significantly higher in the number than that of direct concentration and SPE with C18 as the adsorbent. Six peptides were confirmed with their corresponding synthetic reference standards by comparing their retention time, high resolution MS/MS spectra, and NMR spectroscopic studies. Parallel reaction monitoring integrated with the internal standard method was utilized to quantify identified peptides with concentrations ranging from 1.14 to 10.25 ng mL-1, and prediction results of bioactivity comprising antioxidation or ACE inhibitors were obtained. These discoveries were conducive to understanding the versatility benefit of Baijiu and paved the way to study the interaction between peptides and volatile substances.


Giant enterolith in ileal diverticulum following ileoplastic bladder augmentation.
When adhesions, internal hernias, malignant intra- and retro-peritoneal neoplasms are excluded in patients presenting with new onset constipation and abdominal mass appearance after previous abdominal surgery, other causes must be considered. Giant enteroliths formed within ileal diverticula in the site of small bowel anastomosis may extrude and produce a palpable abdominal lump. Recent experience with such a patient is the basis of this report. Ileal diverticula with interior enteroliths may be suspected in patients presenting with an abdominal lump following previous small bowel resection. Open or laparoscopic assisted surgical resection of the involved segment is the treatment of choice.


Correlation of biofilm expression types of Staphylococcus epidermidis with polysaccharide intercellular adhesin synthesis: evidence for involvement of icaADBC genotype-independent factors.
IcaADBC-encoded proteins mediate synthesis of the intercellular polysaccharide adhesin (PIA), which is essentially involved in Staphylococcus epidermidis biofilm formation. Seventy S. epidermidis isolates were investigated for their ability to form biofilm and synthesize PIA in different growth media including trypticase soy broth obtained from Becton Dickinson (TSBBBL), or Oxoid (TSB(OXOID)), and TSB(OXOID) supplemented with 0.5% N-acetylglucosamine, and for the presence of icaADBC. Dependent on the medium used (TSB(BBL) or TSB(OXOID)), the isolates exhibited a differential expression of PIA and biofilm formation, with 51 (72.85%) and 34 (48.57%) being biofilm positive, respectively. Using these growth media four different expression phenotypes were differentiated: similar quantities of biofilm formation in both TSBBBL and TSB(OXOID) (11 isolates, type A), significantly reduced biofilm expression in TSB(OXOID) compared to TSB(BBL) (23 isolates, type B), biofilm negative in TSB(OXOID) but biofilm producing in TSB(BBL) (17 isolates, type C) and biofilm negative in both media (19 isolates, type D). For all strains a biofilm-positive phenotype in a specific medium was closely linked to expression of PIA in that medium. All but one strain of expression type A-C and 7/19 expression type D strains were icaADBC positive. On the basis of restriction fragment length polymorphisms, the isolates were classified into two main icaADBC genotypes. There was no association between the observed biofilm-expression types and a defined icaADBC genotype. In the biofilm-negative S. epidermidis 5179, isolated from a ventriculo-atrial shunt infection, the insertion of IS257 interrupted the transcription of icaADBC, resulting in a PIA- and biofilm-negative phenotype. In all other icaADBC-positive, biofilm-negative isolates no major alterations of the icaADBC gene locus were identified. Obviously, expression of icaADBC, PIA synthesis and biofilm formation are integrated into a complex regulatory network involving other determinants independent of icaADBC genotype. Inactivation of icaADBC by IS elements is apparently a rare cause of a biofilm-negative phenotype in clinical S. epidermidis isolates.


Upper and Apparent Lower Critical Solution Temperature Branches in the Phase Diagram of Polymer:Small Molecule Semiconducting Systems.
Solution-processable semiconducting materials are complex materials with a wide range of applications. Despite their extensive study and utility, their molecular interactions as manifested, for example, in phase behavior are poorly understood. Here, we aim to understand the phase behavior of conjugated systems by determining phase diagrams spanning extensive temperature ranges for various combinations of the highly disordered semiconducting polymer (PTB7-Th) with crystallizable (IT-M and PC61BM) and noncrystallizable (di-PDI) small molecule acceptors (SMAs), with polystyrene as an amorphous control, a nonsemiconducting commodity polymer. We discover that the apparent binodal of the studied blends frequently consists of an upper critical solution temperature (UCST) and lower critical solution temperature (LCST) branch, exhibiting a sharp kink where the branches join. Our work suggests that phase diagrams might be a probe in combination with sophisticated models to understand the complexity of semiconducting materials, including microstructure and molecular interactions.


Sociotropy/autonomy and vulnerability to specific life events in patients with unipolar depression and bipolar disorders.
Followed samples of unipolar and bipolar patients for a 6-month period, with independent assessment of symptoms and life events. Patients were initially categorized into subtypes using Beck's Sociotropy/Autonomy Scale, with the prediction that onset or exacerbation of symptoms, as well as more total symptoms, would occur for sociotropic individuals experiencing more negative interpersonal events than achievement events, and for autonomous-achievement patients experiencing more achievement events than interpersonal events. Results were confirmed for unipolars, indicating that the course of disorder was associated with the occurrence of personally meaningful life events, but not for bipolars. Further research is recommended to examine whether the effect is equally robust for both subtypes of unipolars, whether longer study duration may be required for bipolars, and whether a cognitive self-schema mechanism may account for the specific vulnerability to a subset of stressful events.


Exposure to ozone enhances antigen-presenting activity concentration dependently in rats.
The effect of ozone (O(3)) on the symptoms of allergic asthma and the mechanisms underlying have not yet been fully elucidated. Antigen presentation is one of the factors contributing to the allergic reaction. Therefore, we investigated the effects of repeated exposure to O(3) on antigen-presenting (AP) activity, on the expression of cell-surface molecules associated with antigen presentation (Ia, B7.1, B7.2 and CD11b/c) in bronchoalveolar lavage cells (BAL cells), and on allergic asthma-like symptoms. Rats were exposed to 0.3, 0.56, 1ppm O(3) or filtered air for a 3-day period every 2 weeks, this was replicated three times. AP activity was assessed by measuring antigen-specific T-cell proliferation; and the expression of cell-surface molecules, by flow cytometry. Rats were also made to inhale aerosolized 1% ovalbumin (OVA) or saline for 10min post-exposure to O(3), and allergic asthma-like symptoms were measured by determining the increase in enhanced pause (Penh), which correlates well with lung resistance. O(3) increased both AP activity and expression of Ia and costimulatory molecules in BAL cells concentration dependently. It also increased lung resistance, and the increase in lung resistance after O(3) exposure was significantly higher in the OVA-inhaled group than in the saline-inhaled group. The present results show that O(3) increased AP activity concentration dependently and suggest that O(3) might aggravate allergy symptoms by enhancing AP activity.


Long-term follow-up of patients with IgA nephropathy treated with prednisolone and cyclophosphamide therapy.
Analyses of selected cases suggest that immunosuppressive treatment could reduce proteinuria and delay the progression of immunoglobulin A nephropathy (IgAN). The aim of this study was to examine the long-term effectiveness of this therapy on the clinical course of IgAN. We also examined the relationship between the efficacy of the treatment and the suppression of the serum immunoglobulin level. Eighteen patients who were observed for more than 2 years after prednisolone and cyclophosphamide therapy were enrolled in this study. Their clinical and laboratory characteristics were recorded for 2-18 years (mean 7.8 +/- 5.7 years). Of the 18 patients, 13 had remission of proteinuria. We observed the subsequent development of proteinuria in four patients. Fourteen patients had remission of hematuria, with five patients experiencing subsequent relapse of hematuria. The mean time from the treatment to the relapse of proteinuria or hematuria was 5.8 years. Serum immunoglobulins were suppressed by the combination therapy. Serum IgG and IgM recovered 6 months after the treatment, whilst the suppression of serum IgA lasted for 4 years. We found a positive correlation between the serum IgA level and the degree of proteinuria. This study indicates that long-term follow-up is essential in order to prove the long-term benefit of immunosuppressive therapy in patients with IgAN. Careful monitoring of the serum IgA level may be useful in the follow-up of patients with IgAN, especially when they are treated with immunosuppressive agents.


The anion exchanger as an osmolyte channel in the skate erythrocyte.
Regulatory volume decrease occurs in fish erythrocytes by the release of osmolytes via a channel involving the anion exchanger (AE), also known as band 3. This review focuses primarily on work done on red blood cells from the skate (Raja erinacea) to further understand the activation, regulation and identification of this osmolyte channel. A model is proposed in which the reduction in intracellular ionic strength that occurs with increased cell volume may change the interaction between AE dimers and other cytoplasmic proteins (band 4.1 and ankyrin), promoting the formation of a tetrameric osmolyte channel. Phosphorylation by two tyrosine kinases, p72syk and p56lyn, is linked to this oligomerization. The skate AE has been recently cloned, resulting in three different isoforms, one of which, skAE1, when expressed in Xenopus oocytes, demonstrates taurine transport.


Optimization of a liquid chromatography method for the analysis of related substances in daclatasvir tablets using design of experiments integrated with the steepest ascent method and Monte Carlo simulation.
Analytical method for the determination of related substances (RS) in Daclatasvir tablets was optimised using quality by design (QbD) approach. Seven degradants (each more than 1.0%) generated during oxidation study, adversely affected the selectivity of the method. Coelution of the degradant peaks with API and known impurities, suggested failure in developing a stability indicating method. To overcome the shortcomings and develop a robust method, QbD principles were incorporated. Resolution was the critical quality attribute (CQA) and buffer pH, column oven temperature, gradient slope and flow rate were the critical method variables (CMVs) studied through design of experiments (DoE). Discovery of an unknown impurity (named as impurity D, about1.0%) was a key finding from this DoE study. The most crucial responses viz. Resolution between impurity D and the main peak and resolution between the main peak and impurity E demanded contradictory pH requirements. To select the right pH, responses were prioritised and eventually to attain the desired resolution between Daclatasvir and impurity E the value for pH was fixed to 3.0. Next, to improve resolution between impurity D and Daclatasvir, method of steepest ascent was applied to locate an apt value for column oven temperature. Accordingly, experiments were performed at different temperatures along the path of rapid increase in response. Finally, at 45 °C (pH :3.0), both the critical pairs were well resolved. The global optimum was determined through a Response surface methodology (RSM) design with pH and column oven temperature as critical factors. pH 3.0, column oven temperature 44 °C, % MP. B 45% and flow rate 1.0 mL min-1 was found to be the optimum condition. Further, the design space was complimented by establishment of a robust zone through Monte Carlo simulation and capability analysis. An analytical control strategy (ACS) was set up to ensure that the method repeatedly meets its acceptance criteria. The optimised method was successfully validated within the factor ranges mentioned in the ACS. Despite various intricacies, the QbD approach facilitated systematic optimisation of a stability indicating robust method.


Sound waves for solving the problem of recrystallization in cryopreservation.
Organ biobanking is the pending subject of cryopreservation. Although the problem is multifaceted, advances in recent decades have largely related it to achieving rapid and uniform rewarming of cryopreserved samples. This is a physical challenge largely investigated in past in addition to cryoprotectant toxicity studies, which have also shown a great amount of advancement. This paper presents a proof-of-principle, based on the nematode Caenorhabditis elegans, of a technology capable of performing such a function: high intensity focused ultrasound. Thus, avoiding the problem of recrystallization, this worm, in its adult state, preserved at - [Formula: see text], has been systematically brought back to life after being heated with High Intensity Focused Ultrasound (HIFU) waves. The great advantage of this technology is that it is scalable; in addition, rewarming can be monitored in real time by MRI thermography and can be controlled by acoustic interferometry. We anticipate that our findings are the starting point for a possible approach to rewarming that can be used for cryopreservation of millimeter scale systems: either alone or in combination with other promising ways of heating, like nanowarming or dielectric heating, the present technology provides new ways of solving the physical aspects of the problem of recrystallization in cryopreservation, opening the door for the long-term storage of larger samples.


Obesity and beta-blockers: influence of body fat on their kinetics and cardiovascular effects.
Beta-blockers are among the most widely used antihypertensive drugs. They differ from each other in regard to several factors such as: beta-agonist activity, beta 1-selectivity and solubility. Aim of this work was to evaluate the influence of obesity on the kinetics and the antihypertensive effect of two Beta-blockers with different solubility such as: the water-soluble, atenolol and the liposoluble, metoprolol. The study was carried out according to an open randomized cross-over design. Eight obese hypertensive patients, after a two week washout period, were randomly allocated to a four week treatment. After a two week intermediate washout period, each patient switched to the other treatment for an additional four week period. On the first and the last day of each treatment the subjects were hospitalized to collect blood samples for the assay of the two drugs and to measure cardiovascular parameters. Obesity does not exert any effect on the kinetics of the water-soluble beta-blocker, atenolol, while markedly interferes with that of the liposoluble, without any apparent influence on its anti-hypertensive effect. These findings extend to obese hypertensives the concept that the plasma concentrations of beta-blocking agents are not reliable predictors of their therapeutic effect.


"I think it is woven through me, and sadly that means it is woven through our family life": the experiences and support needs of mothers with eating disorders.
Eating disorders may disrupt parenting, and there is evidence to suggest that the children of parents with eating disorders are at greater risk for the development of eating disorders themselves. This study sought to broaden and extend current understandings of the lived experiences of mothers who have eating disorders. A qualitative study using thematic analysis was conducted. Eighteen mothers living in the UK, USA, and Australia took part in semi-structured online interviews. Participants were mothers to at least one child aged two years or older, had received a lifetime diagnosis of one or more eating disorders, and reported experiencing symptoms since becoming a parent. Data were analysed using an inductive approach to reflexive thematic analysis. Four major themes, each with subthemes, were identified. These were: parenthood as a double-edged sword; the eating disorder impacts on parenting; blame and burden; and seeking support. The lived experiences of mothers indicate a complex relationship between eating disorders and parenthood. While parenting can impact eating disorders, eating disorders can also impact parenting, in a range of ways that extend beyond the domains of food, eating and the body. There is a pressing need for the development of specialised, non-judgemental support options for parents with eating disorders and their families.


Vectors based on autonomous parvoviruses: novel tools to treat cancer?
Autonomous parvoviruses are small nuclear-replicating DNA viruses. The rodent parvoviruses usually are non- or weakly pathogenic in adult animals, bind to surface receptors which are expressed on most cells, and do not appear to integrate into host chromosomes during either lytic or persistent infections. Interestingly, malignant transformation of the target cells was often found to correlate with an increase in their capacity for amplifying and/or expressing the incoming parvoviral DNA, and is associated with oncolysis, i.e., the selective killing of the infected tumor cells. Moreover, the closely related parvoviruses MVM, H-1 and LuIII efficiently infect human cell lines. This finding makes these parvoviruses promising candidate vectors for therapies that require transient expression of a transduced gene. In particular, parvoviruses may be suitable to target and kill tumor cells and simultaneously deliver appropriate transgenes, e.g., genes coding for immuno-stimulatory factors. Pilot experiments performed in animals to assess whether parvovirus-based vectors carrying the interleukin 2 (IL-2) cytokine gene have reinforced anti-cancer capacity showed that these recombinant viruses suppressed tumor formation more efficiently than viruses devoid of a transgene. Strong anti-cancer effects of recombinant parvoviruses expressing interferon gamma-inducible protein 10 (IP-10) and monocyte chemotactic protein 3 (MCP-3) were also observed against established hemangiosarcomas and melanomas in immuno-competent mice, respectively. Altogether, these data illustrate the enormous potential of recombinant autonomous parvoviruses as anti-tumor agents and give hope of using them against human cancer.


Event-related potentials associated with second language learning in children.
In this study, we investigated learning-related changes in auditory event-related potentials (ERPs) of Finnish-speaking 3-6-year-old children caused by learning French language. Using an oddball paradigm, ERPs to sounds of French language were recorded in the two groups of healthy children: those who were learning French (experimental group) and those who were not learning any foreign language (control peers). When the children from the experimental group were exposed to the foreign language, they automatically developed French-specific memory traces that helped them to discriminate, categorize, and pronounce utterances of the new language as indicated by the mismatch negativity (MMN) component of the ERPs in a previous study. We found that the learning process was also reflected by changes in P3a and late difference negativity (LDN) responses. Unlike MMN and P3a, the LDN has been discovered relatively recently and its functional role remains unclear. Similarly, as the MMN magnitude increased during the learning process, an increase of the P3a (known to reflect the involuntary attention switching toward deviant stimuli) and LDN amplitudes was observed. The ERPs of the control peers did not change significantly over the test period. When phonemes of a foreign language are learned, this process is accompanied with the increase in the MMN, P3a, and LDN amplitudes in children. Though the functional significance of LDN remains to be further investigated, our results support its possible link to reorienting processes following distraction.


Ultrafast chirality: the road to efficient chiral measurements.
Today we are witnessing the electric-dipole revolution in chiral measurements. Here we reflect on its lessons and outcomes, such as the perspective on chiral measurements using the complementary principles of "chiral reagent" and "chiral observer", the hierarchy of scalar, vectorial and tensorial enantio-sensitive observables, the new properties of the chiro-optical response in the ultrafast and non-linear domains, and the geometrical magnetism associated with the chiral response in photoionization. The electric-dipole revolution is a landmark event. It has opened routes to extremely efficient enantio-discrimination with a family of new methods. These methods are governed by the same principles but work in vastly different regimes - from microwaves to optical light; they address all molecular degrees of freedom - electronic, vibrational and rotational, and use flexible detection schemes, i.e. detecting photons or electrons, making them applicable to different chiral phases, from gases to liquids to amorphous solids. The electric-dipole revolution has also enabled enantio-sensitive manipulation of chiral molecules with light. This manipulation includes exciting and controlling ultrafast helical currents in vibronic states of chiral molecules, enantio-sensitive control of populations in electronic, vibronic and rotational molecular states, and opens the way to efficient enantio-separation and enantio-sensitive trapping of chiral molecules. The word "perspective" has two meanings: an "outlook" and a "point of view". In this perspective article, we have tried to cover both meanings.


Unveiled central hypoventilation after tracheotomy in anti-IgLON5 disease: a case report.
Anti-IgLON5 disease is a recently described entity that has been associated with neurological symptoms and sleep disturbances including sleep breathing disorders. Sleep stridor as well as obstructive and less often central sleep apnea have been reported but rarely needing ventilation on tracheotomy. We report the case of a patient in whom obstructive sleep apnea with secondary development of dysphagia and recurrent aspiration pneumonia led to the diagnosis of anti-IgLON 5 disease. Acute respiratory failure due to laryngospasm required intubation and eventually tracheotomy. Yet hypoventilation persisted, and polysomnography demonstrated central sleep apnea alternating with sleep-related tachypnea. Nocturnal ventilation was thus reintroduced. The association of obstructive sleep apnea with dysphagia is a potential red flag for anti-IgLON5 disease, which remains an overlooked diagnosis. Breathing disorders can be complex in this context, with a mixed obstructive and central pattern whose central component can be unveiled after tracheotomy. This highlights the importance of closely monitoring sleep and respiration even after tracheotomy. Tankéré P, Le Cam P, Folliet L, et al. Unveiled central hypoventilation after tracheotomy in anti-IgLON5 disease: a case report. J Clin Sleep Med. 2023;19(9):1701-1704.


Oral nerve sheath myxoma. Report of a case with findings of ultrastructural and immunohistochemical studies.
The first Asian case of oral nerve sheath myxoma is presented, along with the findings of light and electron microscopy, and immunohistochemistry. Ultrastructurally, intracytoplasmic microfilaments and convoluted basement membrane were identified in the tumor cells. Immunohistochemically, S-100 protein and neuron-specific enolase were positive in the tumor cells. These findings may indicate that the tumor is a lesion of Schwann-cell origin. The literature on oral nerve sheath myxoma is also reviewed and discussed.


Mode of delivery and development of atopic disease during the first 2 years of life.
It has been hypothesized that cesarean delivery might have an impact on the development of atopic diseases because of its gut flora modulating properties. In the present study, we analysed the association between cesarean delivery and atopic diseases using data of 2500 infants enrolled in the LISA-Study, a German prospective multicenter birth cohort study. Data on symptoms and physician-diagnosed atopic diseases were gathered by questionnaires shortly after birth and at infant's age 6, 12, 18, and 24 months. In addition, sensitization to common food and inhalant allergens was assessed by measuring specific immunoglobulin E (IgE) using the CAP-RAST FEIA method at the age of 2 yr. Confounder-adjusted odds ratios (aOR) with 95% confidence intervals (CI) were calculated by multiple logistic regression. We found a positive association between cesarean delivery and occurrence of at least one episode of wheezing [aOR 1.31 (95% CI 1.02-1.68)] and of recurrent wheezing [1.41 (1.02-1.96)] during the first 2 yr of life. Furthermore, effect estimates for allergic sensitization defined as at least one specific IgE >/=0.70 kU/l against any allergen [1.48 (0.98-2.24)], against food allergens [1.64 (1.03-2.63)], and against inhalant allergens [1.75 (0.98-3.12)] were increased. Symptoms of atopic dermatitis [1.21 (0.92-1.59)], physician-diagnosed atopic dermatitis [1.04 (0.79-1.39)], and symptoms of allergic rhinoconjunctivitis [1.40 (0.80-2.44)] were only marginally increased in children delivered by cesarean section. In conclusion, our results suggest that cesarean delivery may be an additional risk factor for wheezing and allergic sensitization at least to food allergens up to the age of 2 yr. This should be considered when cesarean section is done for other than medical reasons.


Evaluation of hearing organ in patients with Turner syndrome.
Hearing loss and middle ear diseases are often reported in some of Turner patients. In most of the reports hearing organ was evaluated with the use of subjective methods. The aim of the work was subjective and objective evaluation of hearing organ with an attempt to set the correlation between the results and the genotype of the patients with Turner syndrome (Ts). 51 Ts patients aged 14.3 years on average. There were 29 girls with monosomy X and 22 having mosaicism. A detailed medical history was taken in each case with attention given to the hearing loss risk factors. Physical ENT examination, hearing evaluation: pure tone audiometry, impedance audiometry, distortion products otoacoustic emissions (DPOAEs), brain auditory evoked potentials (BAEP). The control group consisted of 30 healthy patients. Recurrent acute otitis media was reported by 19.6% of Ts patients. Pure tone audiometry was improper in 36.3% ears; conductive hearing loss was present in 11.7% ears, mixed hearing loss in 5.9% ears and the moderate sensorineural hearing loss in 18.6% ears. Impedance audiometry was impaired in 14.7% of the cases. DPOAE disturbances were present in 41.4% of Ts patients, BAEP was improper in 52.0%. The percentage of the disturbances in DOPAEs and in BAEP in patients with mosaicism was 45.4 and 40.9% while in patients with monosomy 68.9 and 62%. Ts patients present predisposition to hearing disturbances. The disturbances seem to be connected with middle ear infections and with sensorineural hearing losses. Hearing loss in Ts women is not clinically apparent in most of the cases; this fact reflects the need of early evaluation and further monitoring of hearing organ in those patients. Sensorineural hearing loss seems to prevail in patients with 45,X genotype, so perhaps attention should be paid to this subgroup of Ts patients.


Cross clade prophylactic and therapeutic efficacy of polyvalent equine immunoglobulin F(ab')2 against highly pathogenic avian influenza H5N1 in mice.
Highly pathogenic avian influenza H5N1 virus (HPAI H5N1) has the potential to cause a new pandemic, which may lead to disasters in the world. However, we cannot predict the HPAI H5N1 strain that might cause the pandemic. Therefore, broad-spectrum prophylactic or therapeutic preparations for containment of a possible future pandemic are urgently needed. Polyvalent equine immunoglobulin F(ab')2 may be a promising candidate. We prepared four pepsin digested immunoglobulin F(ab')2 from the horses immunized with purified VNH5N1-Puerto Rico/8/34 (PR8)/CDC-RG (VNRG, Clade 1), A/Indonesia/05/2005(H5N1)-PR8-IBCDC-RG2 (INRG, Clade 2.1), and A/Anhui/01/2005(H5N1)-PR8-IBCDC-RG5 (AHRG, Clade 2.3.4) and PBS (negative control), respectively. The protective effect of the monovalent or polyvalent F(ab')2 against A/Ostrich/SZ/097/04 (clade 0) infection was determined by cytopathic effect (CPE) in cultured Madin-Darby canine kidney (MDCK) cells. The prophylactic and therapeutic efficacy of the polyvalent F(ab')2 was further evaluated by observing survival, weight loss and viral load when the polyvalent F(ab')2 was introduced into mice one day prior to-, three days post-lethal challenge with A/Ostrich/SZ/097/04. The half neutralization doses (ND50) of purified monovalent equine F(ab')2 prepared by the VNRG, INRG or AHRG-immunized horses and polyvalent one against A/Ostrich/SZ/097/04 were 320, 1280, 1280 and 2560 in cultured MDCK cells, respectively. 10 μg polyvalent F(ab')2 could completely protect mice infected with 100 half lethal doses (LD50) of A/Ostrich/SZ/097/04 in preventive settings. In therapeutic settings, even when injected three days post lethal infection, mice were still completely protected, although 200 μg of polyvalent F(ab')2 was required. Our work has provided experimental supports for testing the broad-spectrum protective efficacy of polyvalent equine immunoglobulin F(ab')2 for the future large trials.


Knee Valgus during Jump Landing Is Related to the Inaccuracy of Knee Position Recognition in Healthy Young Women.
Knee valgus during jump landing is a cause of knee injuries during sports activities. Body recognition is important for maintaining the knees and other body parts in their proper positions. The aim of this study was to investigate whether knee valgus during jump landing in healthy young women is related to the inaccuracy of recognition of bilateral knee positions in the squatting position. In 39 healthy young women, the degree of knee valgus was evaluated during the drop vertical jump test using the ratio of the knee separation distance to the ankle separation distance. The accuracy of recognition of bilateral knee positions in the squatting position was evaluated by having the blindfolded subjects indicate with their index fingers the subjective positions of their bilateral patellae by placing their fingers on a horizontal bar positioned in front of them 3 cm below the navel. The difference ratio of the recognized distance to the actual distance between the bilateral patellae was measured as an inaccuracy index. The degree of knee valgus during the drop vertical jump test was positively correlated with the degree of inaccuracy of the recognized knee position with the knees in the neutral position (r=0.358, P=0.025). In healthy young women, knee valgus during jump landing was significantly correlated with the inaccuracy in knee position recognition in the squatting posture. This finding suggests that the assessment of knee position recognition in the squatting position could be useful as a screening tool for preventing knee injuries on jump landing during sports activities.


(Nitrato-κO)(2,2':6',2''-terpyridine-κ3 N,N',N'')palladium(II) nitrate.
The title complex, [Pd(NO3)(C15H11N3)]NO3, comprises a cationic PdII complex and a nitrate anion. In the complex, the PdII cation is four-coordinated in a distorted square-planar coordination geometry defined by the three N atoms of the tridentate 2,2':6',2''-terpyridine ligand and one O atom from the NO3 - anion. In the crystal, the complex mol-ecules are stacked in columns along the a axis being connected by π-π stacking [closest inter-centroid separation between pyridyl rings = 3.878 (3) Å]. The connections between columns and anions to sustain a three-dimensional architecture are C-H⋯O hydrogen bonds.


Trajectories of parenting behavior and maternal depression.
This study investigated trajectories of maternal parenting behavior across the infants' first 18 months of life in relation to maternal depression. Furthermore, predictors of the quality of the mother-infant relationship at 18 months were examined. Participants consisted of three types of mother-infant dyads: mothers with comorbid depression and anxiety (n=19), mothers with depression (n=7) and nondepressed mothers (n=24). Maternal behaviors and the quality of relationship were rated on a global scale (NICHD) from video-taped mother-infant interactions. Maternal behaviors rated at six, 12 and 18 months were collapsed into a composite variable maternal style. The quality of the relationship captured as dyadic mutuality was rated at 18 months. Comorbid and depressed mothers showed lower quality in maternal style compared with the nondepressed mothers at six months. Over the follow-up the comorbid mothers were lower in maternal style compared to the nondepressed mothers, but the comorbid mothers increased significantly in maternal style despite elevated depression symptoms. Mean maternal style and infant cognitive skills predicted the quality in relationship at 18 months suggesting that the mother-toddler relationship depends on contributions from the mother and the child. Higher growth in maternal style despite of depression symptoms among comorbid mothers was interpreted against the background that the majority of the comorbid mother-infant dyads received several treatments.


Predictors of Readmission Following Treatment for Traumatic Hemothorax.
Retained-hemothorax after trauma can be associated with prolonged hospitalization, empyema, pneumonia, readmission, and the need for additional intervention. The purpose of this study is to reduce patient morbidity associated with retained-hemothorax by defining readmission rates and identifying predictors of readmission after traumatic hemothorax. The Nationwide Readmission Database for 2017 was queried for patients with an index admission for traumatic hemothorax during the first 9 mo of the year. Deaths during the index admission were excluded. Data collected includes demographics, injury mechanism, outcomes and interventions including chest tube, video-assisted thoracoscopic surgery, and thoracotomy. Chest-related readmissions (CRR) were defined as hemothorax, pleural effusion, pyothorax, and lung abscess. Univariate and multivariate analysis were used to identify predictors of readmission. There were 13,903 patients admitted during the study period with a mean age of 53 ± 21, 75.2% were admitted after blunt versus 18.3% penetrating injury. The overall 90-day readmission rate was 20.8% (n = 2896). The 90-day CRR rate was 5.7% (n = 794), with 80.5% of these occurring within 30 d. Of all CRR, 62.3% (n = 495) required an intervention (chest tube 72.7%, Thoracotomy 26.9%, video-assisted thoracoscopic surgery 0.4%). Mortality for CRR was 6.2%. Predictors for CRR were age >50, pyothorax or pleural effusion during the index admission and discharge to another healthcare facility or skilled nursing facility. Majority of CRR after traumatic hemothorax occur within 30 d of discharge and frequently require invasive intervention. These findings can be used to improve post discharge follow-up and monitoring.


Sab mediates mitochondrial dysfunction involved in imatinib mesylate-induced cardiotoxicity.
Imatinib mesylate is an effective treatment for chronic myelogenous leukemia and gastrointestinal stromal tumors. Although imatinib mesylate is highly tolerable, it has been implicated in severe congestive heart failure in mouse models and patients. A hallmark of imatinib mesylate-induced cardiotoxicity is mitochondrial dysfunction. The mitochondrial scaffold Sab has been implicated in facilitating signaling responsible for mitochondrial dysfunction in a c-Jun N-terminal Kinase (JNK)-dependent manner. We examined the impact of Sab-mediated signaling on imatinib mesylate cardiotoxicity in H9c2 rat cardiomyocyte-like cells. Silencing Sab increased the LD50 of imatinib mesylate 4-fold in H9c2 cells. Disrupting Sab-mediated signaling prevented imatinib mesylate-induced apoptosis as well. Knockdown of Sab or inhibition with a small peptide prevented oxidative stress, which was indicated by decreased reactive oxygen species production, lipid peroxidation, and protein carbonylation. Further, inhibition of Sab-related signaling partially rescued deficits in mitochondrial respiration, ATP production, and membrane potential in imatinib mesylate-treated H9c2 cells. Conversely, over-expression of Sab in H9c2 cells increased the cardiotoxicity of imatinib mesylate in vitro decreasing the LD50 over 4-fold. Sab expression was induced in H9c2 cells following cardiovascular-like stress in an AP-1 dependent manner. These data demonstrate that imatinib mesylate influences mitochondrial signaling leading to mitochondrial dysfunction and cardiotoxicity.


A phase II randomized trial of sodium oligomannate in Alzheimer's dementia.
Sodium oligomannate (GV-971), a marine-derived oligosaccharide, is a novel agent that may improve cognition in AD patients. The 24-week multicenter, randomized, double-blind, placebo parallel controlled clinical trial was conducted in AD in China between 24 October 2011 and 10 July 2013. The study included a 4-week screening/washout period, followed by a 24-week treatment period. Patients were randomized in a 1:1:1 ratio to receive GV-971 900 mg, 600 mg, or placebo capsule in treatment period, respectively. The primary outcome was cognitive improvement as assessed by changes in Alzheimer's Disease Assessment Scale-cognitive subscale 12-item (ADAS-cog12) scores from baseline to week 24. The secondary efficacy outcomes included CIBIC-Plus, ADCS-ADL, and NPI at 24 weeks after treatment compared with baseline. A subgroup study was assessment of the change in cerebral glucose metabolism by fluorodeoxyglucose positron emission tomography measurements. Comparing with the placebo group (n = 83, change - 1.45), the ADAS-cog12 score change in the GV-971 600-mg group (n = 76) was - 1.39 (p = 0.89) and the GV-971 900-mg group (n = 83) was - 2.58 (p = 0.30). The treatment responders according to CIBIC-Plus assessment were significantly higher in the GV-971 900-mg group than the placebo group (92.77% vs. 79.52%, p < 0.05). The GV-971 900-mg subgroup showed a lower decline of cerebral metabolic rate for glucose than the placebo subgroup at the left precuneus, right posterior cingulate, bilateral hippocampus, and bilateral inferior orbital frontal at uncorrected p = 0.05. The respective rates of treatment-related AEs were 5.9%, 14.3%, and 3.5%. GV-971 was safe and well tolerated. GV-971 900 mg was chosen for phase III clinical study. ClinicalTrials.gov, NCT01453569 . Registered on October 18, 2011.


Factors that determine the occurrence of reperfusion arrhythmias.
The determinants of reperfusion arrhythmias were investigated in 63 open-chest dogs undergoing a 25-minute coronary artery occlusion followed by reperfusion. Heart rate correlated positively with the occurrence of reperfusion ventricular tachycardia (VT) and ventricular fibrillation (VF). Collateral flow during ischemia (radioactive microspheres) exhibited a strong negative correlation with the incidence of both VT and VF upon reperfusion. Importantly, a sensitive coupling was present, whereby small differences in flow were associated with large differences in rhythm disorders. The rise in intramyocardial CO2 tension (another index of severity of ischemia) was greater in dogs exhibiting reperfusion VT (p less than 0.001) and VF (p less than 0.08); however, this variable was significantly correlated with collateral flow (r = -0.57, p less than 0.01). The size of the occluded coronary bed, determined by postmortem perfusion, was not consistently related to VT; within a given range of occluded bed sizes, the incidence of VT was inversely related to collateral flow. Thus, reperfusion-induced VT is relatively independent of the size of the occluded bed, and is determined primarily by the degree of myocardial hypoperfusion. In contrast, VF did not develop with occluded beds less than 25% of the left ventricular mass; above this critical occluded bed size, its incidence was inversely related to collateral perfusion. Reperfusion VF is therefore determined by the association of a large occluded bed with a poor collateral flow. This study identifies and systematically analyzes three major determinants of reperfusion arrhythmias: (1) the severity of antecedent ischemia, estimated either from the degree of flow reduction or the rise in intramural CO2 tension; (2) the amount of ischemic/reperfused myocardium; and (3) the heart rate. In addition to conceptual interest and clinical implications, the findings have important implications for the design of future studies aimed at evaluating antiarrhythmic interventions in experimental models.


Intraneuronal advanced glycation endproducts in presenilin-1 Alzheimer's disease.
The most frequently mutated gene resulting in dominantly inherited Alzheimer's disease is presenilin-1. We have used antibodies against advanced glycation endproducts (AGE) in brain tissue sections of four patients with three different presenilin I mutations. Accumulation of intracellular AGE was observed in 75-95% of pyramidal neurons in patients with presenilin-1 mutations, far exceeding the percentage of presenilin-1-, tau- or ubiquitin-positive neurons. This high level of AGE-modified proteins in vulnerable neurons is most likely explained by higher levels of their precursors (reactive (di)carbonyl products) or a slower turnover of the participating proteins. These conditions of carbonyl stress may contribute to increased neuronal dysfunction and vulnerability leading to the early disease onset.


Low protein with high amino acid diets improves the growth performance of yellow feather broilers by improving intestinal health under cyclic heat stress.
This study investigated the effects of a low-protein diet supplemented with high lysine, methionine, and threonine levels on production performance, antioxidant capacity, intestinal morphology, duodenum gene expression and intestinal microorganisms in yellow feather broilers under cyclic heat stress. A total of 162 yellow feather broilers that were 42 d of age were selected and randomly divided into three groups: the control group (CONT, CP 16%), heat stress group (HS, CP 16%), and heat stress with low protein and high amino acid group (HS_LP, CP 14.5%). Following 14 d heating stress period, the HS_LP group showed no significant effect on production performance compared with the HS group. After a 28 d feeding trial, compared with the HS group, the HS_LP group significantly reduced feed: gain at 15-28 d (P < 0.05), had a tendency to reduce feed: gain at 1-28 d (P = 0.056). Compared with the HS group, the serum catalase enzyme activity tended to be higher (P = 0.067), and liver glutathione peroxidase enzyme activity was significantly decreased (P < 0.05) in the HS_LP group. Other antioxidant capacity indexes were not significantly different between the two groups (P > 0.05). Duodenum (P < 0.05) and ileum (P < 0.05) villus height were significantly increased, duodenum villus height: crypt depth ratio (P < 0.05) and jejunum crypt depth (P < 0.05) were significantly decreased, and jejunum villus height was significantly reduced (P < 0.05) in broilers fed the HS_LP diet. Compared with the HS group, the mRNA level of Claudin-1 was significantly increased (P < 0.05), and had a tendency to increase Occludin (P = 0.060) in the HS_LP group. In addition, the HS_LP group significantly increased Nitrosomonas abundance (P < 0.05) and had a tendency to reduce unidentified_Mollicutes abundance (P = 0.083) at the genus level compared with the HS group. This information is useful to formulate diets that correct the decrease in amino acid consumption associated with the reduced voluntary feed intake of broilers under heat stress.


Fat Test Depression During Chilled Storage of Milk Samples in Plastic Containers for Analysis by the Milko-Tester 1.
Ambiguous depressions in fat tests were observed when milk samples were stored before testing by the Milko-Tester procedure. Fat-test depression was augmented by shipping and by chilling samples at the time of collection in an ice-water mixture. The effect was greatest in samples stored in 1-oz. plastic containers and in partially filled plastic bags. The fat-test depression was not significant in samples stored in glass bottles or in plastic containers receiving no refrigeration during the storage period.


Nicotine-modulated formation of spiroiminodihydantoin nucleoside via 8-oxo-7,8-dihydro-2'-deoxyguanosine in 2'-deoxyguanosine-hypochlorous acid reaction.
Hypochlorous acid (HOCl) is generated by myeloperoxidase of activated neutrophils which kill invading microorganisms, but also cause DNA damage in inflamed tissues. We report here that spiroiminodihydantoin nucleoside (dS), a further oxidized product of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), is formed, in addition to 8-chloro-2'-deoxyguanosine and 8-oxo-dG, by reaction of 2'-deoxyguanosine with HOCl. Presence of low concentrations of nicotine significantly enhanced the yields of these HOCl-modified nucleosides. Our results imply that nicotine may enhance genotoxicity and tissue damage caused by neutrophil activation. dS may also serve as a new biomarker for oxidative DNA damage induced by oxidants such as HOCl.


Desensitization of cholecystokininB receptors in GH3 cells.
Desensitization of the cholecystokinin (CCK) octapeptide (CCK-8)-induced rise in intracellular free calcium concentration ([Ca2+]i) was characterized in GH3 cells, a pituitary tumor cell line, which are known to possess CCKB receptor subtype. The CCK-8-induced [Ca2+]i transient was reduced following the initial application of CCK-8. A similar desensitization of the CCK-8-induced response was observed following the first application of thyrotropin-releasing hormone (TRH). By contrast, the TRH-induced response was not desensitized by the preceding application of CCK-8. Desensitization of the CCK-8-induced [Ca2+]i transient was associated with diminished inositol 1,4,5-trisphosphate formation. The recovery of desensitization of the CCK-8-induced response was delayed by a phosphoserine/phosphothreonine phosphatase inhibitor, calyculin A (100 nM). The responsiveness to CCK-8 was also reduced by phorbol 12,13-dibutyrate (PDBu), and this effect of PDBu was completely abolished by preincubation with staurosporine. Staurosporine significantly attenuated the desensitization caused by preincubation with CCK-8, but this effect was too small to attribute the desensitization to the protein kinase C transduction pathway alone. It is likely that desensitization of CCK receptors involves multiple transduction pathways.


Attractive and healthy-looking male faces do not show higher immunoreactivity.
Previous research has indicated that facial attractiveness may provide cues to the functioning of the immune system. Mating with individuals who have a more effective immune system could lead to a higher reproductive success. Our main aim was to test a possible association between immunoreactivity (stimulated by vaccination) and perceived facial attractiveness and healthiness. We experimentally activated the immune system of healthy men using vaccination against hepatitis A/B and meningococcus and measured levels of specific antibodies (markers of immune system reactivity) before and 30 days after the vaccination. Further, 1 day before the vaccination, we collected their facial photographs that were judged by females for attractiveness, healthiness, and facial skin patches for healthiness. In view of its proposed connection with the functioning of the immune system, we also measured skin colouration (both from the facial photographs and in vivo using a spectrophotometer) and we assessed its role in attractiveness and healthiness judgements. Moreover, we measured the levels of steroid hormones (testosterone and cortisol) and the percentage of adipose tissue, because both are known to have immunomodulatory properties and are related to perceived facial attractiveness and healthiness. We found no significant associations between antibody levels induced by vaccination and perceived facial attractiveness, facial healthiness, or skin healthiness. We also found no significant connections between steroid hormone levels, the amount of adipose tissue, rated characteristics, and antibody levels, except for a small negative effect of cortisol levels on perceived facial healthiness. Higher forehead redness was perceived as less attractive and less healthy and higher cheek patch redness was perceived as less healthy, but no significant association was found between antibody levels and facial colouration. Overall, our results suggest that perceived facial attractiveness, healthiness, and skin patch healthiness provide limited cues to immunoreactivity, and perceived characteristics seem to be related only to cortisol levels and facial colouration.


Parasagittal thalamic knife-cuts and cardiac changes.
Rabbits received knife-cuts either medial or lateral to the mediodorsal nucleus of the thalamus (MD), and had stimulating electrodes implanted in MD. These animals were compared to animals with sham knife-cuts for (1) the cardiac component of the orienting reflex (OR) to novel tone stimuli; (2) differential pavlovian heart rate (HR) conditioning, utilizing tones of different frequencies as CSs and paraorbital shock as the US; and (3) heart rate, blood pressure, electromyographic activity, and respiration changes elicited by electrical stimulation in MD. Medial knife-cuts led to faster habituation of the OR, and to somewhat attenuated HR conditioning, but had no effect on stimulation-elicited changes. Lateral knife-cuts, on the other hand, enhanced the stimulation-elicited HR response, but had little effect on the HR OR. The HR CR was unaffected by lateral cuts during an initial conditioning session, but was attenuated in both knife-cut groups during a second session, compared to sham control animals.


Characterization of the intracellular transport of GluR1 and GluR2 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor subunits in hippocampal neurons.
Little is known about the dynamics of the dendritic transport of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs) to synapses. Here, using virally expressed green fluorescent protein (GFP)-GluR1 and GFP-GluR2 and confocal photobleach techniques we show near real-time movement of these subunits in living cultured hippocampal neurons. GFP-GluR1 fluorescence was widely distributed throughout the extranuclear compartment with no evidence for discrete intracellular stores. GFP-GluR1 transport was predominantly proximal to distal at rates of 0.2-0.4 mum.s-1. GFP-GluR2 fluorescence was more punctate and localized at or close to the plasma membrane. Overall, GFP-GluR2 movement was less dynamic with distinct mobile and immobile pools. Neither activation nor inhibition of surface-expressed N-methyl-d-aspartate receptors or AMPARs had any significant effect on the rates of GFP-GluR1 or GFP-GluR2 dendritic transport. These results demonstrate that GluR1 is constitutively and rapidly transported throughout the neuron. GluR2, on the other hand, is less mobile, with a majority retained in relatively immobile membrane-associated clusters, with approximately 40% showing synaptic co-localization. Furthermore, the transport of both subunits is activity-independent, suggesting that the regulated delivery of AMPARs to the vicinity of synapses is not a mechanism that is involved in processes such as synaptic plasticity.


The air-blood barrier in the human lung. A freeze-fracture study.
The air-blood barrier was studied in replicas of freeze-fractured lung biopsies collected from healthy human subjects. Adjacent pneumocytes display a belt-like network composed of 3-7 superimposed ridges ("fibrils") on the P face and complementary grooves on the E face, i.e., a structure corresponding to a "tight" junction. On the other hand, adjacent capillary endothelial cells show a continuous system of 2-4 membrane foldings. These appear mainly as smooth surfaced crests on the P face; on the E face furrows are seen, at the bottom of which a row of particles is situated. This arrangement suggests a "leaky" type of junction. Discontinuous occluding junctions are located in the pericytic venular segment of the alveolar vessels. The present findings are in agreement with previous physiological and ultrastructural tracer studies locating the main part of the diffusion barrier for small polar solutes and proteins in the alveolar epithelium. Communicating junctions are demonstrated between type I and type II pneumocytes, indicating intercellular cooperation between these cells of common embryonic origin, but which fulfill different functions in the adult. In the endothelium of the non-muscular alveolar vessels communicating junctions are lacking. Desmosomes occur in the epithelium between type I and type II pneumocytes; square arrays of particles characterize the plasma membrane of type I pneumocytes.


Multiple superovulations in N'Dama heifers.
Five N'Dama heifers were superovulated with follicle stimulating hormone (FSH-P or Folltropin) a total of six times each. The superovulations were carried out between ongoing experimental Trypanosoma congolense infections. Twenty-four (80%) of the 30 superovulations had a good ovarian response with 21 (70%) producing an average of 2.7 +/- 0.4 (mean +/- s.e.m.) embryos. The highest embryo production was achieved at the third and fourth superovulation, after which both the number of embryos and their quality declined. The overall pregnancy rate after transfer into Boran (Bos indicus) cow recipients was 50.9%. The uteri of the heifers increased considerably in size throughout the six superovulations which made it difficult to flush some of the animals after the third superovulation. Embryo transfer technology is a useful breeding tool in N'Dama heifers and multiple superovulations can be carried out with success.


[Multiple hypochromic or achromic macules in children and risk of tuberous sclerosis].
To describe in a large paediatric cohort the characteristics of hypopigmented and depigmented (hypochromatic and achromic) macules with no clear diagnosis but potentially evocative of tuberous sclerosis (TS). This was a retrospective multicentre study performed between 2010 and 2017 at a reference centre for rare skin diseases; it included all children consulting for hypochromic and achromic macules. A descriptive analysis was made of the characteristics of macules with no clear diagnosis, enabling them to be classified in three secondary groups: TS certain, TS ruled out, TS uncertain. Of the 3300 children seen during this 7-year period 7,265 were consulting for hypochromic or achromic macules, with no clear diagnosis in 18 cases: 7 girls and 11 boys of median age at 7.21 years (range: 4 months to 16 years and 7 months). The lesions were congenital in 7 cases. The number of macules varied, with over 20 in some cases. The majority were in the form of ash-leaf spots, followed by the oval form. Two children were diagnosed at clinical examination, and 16 underwent it is not examinations, resulting in a diagnosis of certain ST in 6 of these cases. No particular characteristics of the macules appeared to guide the clinical examination towards ST or isolated lesions. Café-au-lait spots were more frequent in the group in which ST was ruled out than in the other two groups: 67% vs. 33% and 33%. Neurologic involvement was more common in children with certain or uncertain ST than in children in whom ST was ruled out (83% and 67% vs. 11%). No identified characteristics of stains enabled the clinical examination to confirm or rule out tuberous sclerosis. Screening for acute any signs of ST is essential. Diagnostic efficacy is enhanced by additional exams.


Pyroptosis of chondrocytes activated by synovial inflammation accelerates TMJ osteoarthritis cartilage degeneration via ROS/NLRP3 signaling.
Synovial inflammation and chondrocyte death have been widely acknowledged as key contributors to the pathological progression of temporomandibular joint osteoarthritis (TMJ-OA), a degenerative joint disease currently lacking definitive treatments. This study aims to understand the regulatory role of chondrocyte pyroptosis in condylar cartilage degradation during TMJ-OA. The levels of cytokines, cartilage degeneration markers, and pyroptotic biomarkers in the synovium and synovial fluid of temporomandibular disorders (TMD) patients were examined. The synovitis, cartilage degradation, and chondrocyte pyroptosis in wild-type and alpha-kinase 1 (ALPK1)-deficient TMJ-OA mice were then compared following monosodium iodoacetate (MIA) induction. Subsequently, we investigated the downstream mechanisms of cytokines- or macrophage supernatants-induced metabolic disorders and pyroptosis in chondrocytes using primary TMJ chondrocytes and ATDC5 chondrocyte cultures. We found a positive correlation between pyroptotic biomarkers and cartilage degradation mediators and cytokines in the synovial fluid of TMD patients. MIA-induced TMJ-OA mice demonstrated significant synovitis, cartilage degradation, and chondrocyte pyroptosis, which were mitigated in ALPK1-deficient TMJ-OA mice, inflammation-restrained mice. Ex-vivo study revealed the contribution of reactive oxygen species (ROS) to inflammation-irritated macrophage supernatants-induced pyroptosis and metabolic disorders in chondrocytes. Targeting NOD-like receptor protein 3 (NLRP3) alleviated cytokines- or ROS-induced pyroptosis and metabolic disorders in chondrocytes by inhibiting caspase-1 activation and interleukin-1β (IL-1β) secretion. Our findings offer novel insight into the role of synovial inflammation-induced chondrocyte pyroptosis in promoting cartilage degradation during TMJ-OA via the ROS and NLRP3 signaling pathway.


Effect of adjusted cuff pressure of endotracheal tube during thyroidectomy on postoperative airway complications: prospective, randomized, and controlled trial.
Postoperative sore throat (POST) is a common complaint after general anesthesia, especially following thyroidectomy. We examined the effect of adjusting the endotracheal tube cuff pressure during thyroidectomy on the incidence of airway complications. Ninety patients scheduled for elective thyroidectomy were randomized into two groups: control (group A, n = 45) and experimental (group B, n = 45). All patients underwent total intravenous anesthesia with propofol and remifentanil. In group A, the cuff pressure was set to 25 cm H2O initially and then monitored continuously without adjustment during thyroidectomy. In group B, the cuff pressure was maintained at approximately 25 cm H2O throughout the operation. The incidences and the severity of POST, hoarseness, dysphagia, and cough were recorded at 2 and 24 h postoperatively. Cuff pressures in group A changed significantly over time (P < 0.05) and were higher than those of group B during thyroidectomy (P < 0.05). The incidences of POST were lower in group B than in group A at 2 and 24 h postoperatively (P < 0.05), and there was a significant difference in the severity of POST at 2 h postoperatively between the two groups. There were no differences in the incidences of hoarseness, dysphagia, and cough between the two study groups (P > 0.05). Adjusting the endotracheal cuff pressure during thyroidectomy decreased the incidence and degree of POST. Intraoperative monitoring and adjustment of the cuff pressure can reduce POST in patients undergoing thyroidectomy.


Discrimination between parkinsonian tremor and essential tremor using artificial neural network with hybrid features.
Essential tremor (ET) and the tremor in Parkinson's disease (PD) are the two most common pathological tremors with a certain overlap in the clinical presentation. The main purpose of this work is to use an artificial neural network to select the best features and to discriminate between the two types of tremors. The features used are of hybrid type obtained from two different algorithms: the statistical signal characterization (SSC) of the signal describing its morphology, and the soft-decision wavelet-decomposition (SDWD) features extracted from the accelerometer and surface EMG signals. The SSC method is used to obtain morphology-based features of the spectrum of the accelerometer and two surface EMG signals. The SDWD technique is used in this work to obtain the approximate spectral representation of both accelerometer and the two surface EMG signals. Two sets of data (training and test) are used in this paper. The training set consists of 21 ET subjects and 19 PD subjects, while the test set consists of 20 ET and 20 PD subjects. A neural network of the type feed forward back propagation has been used to combine best SSC features and best SDWD features of the accelerometer and EMG signals. Efficiency result of 92.5% was obtained using best hybrid features. The artificial neural network has been used successfully to combine two types of features in an automatic discrimination system between PD and ET.


Univariate and Multivariate Base Rates of Score Elevations, Reliable Change, and Inter-Rater Discrepancies in the BRIEF-A Standardization Samples.
The Behavior Rating Inventory of Executive Function-Adult Version (BRIEF-A) is a standardized rating scale of subjective executive functioning. We provide univariate and multivariate base rates (BRs) for scale/index scores in the clinical range (T scores ≥65), reliable change, and inter-rater information not included in the Professional Manual. Participants were adults (ages = 18-90 years) from the BRIEF-A self-report (N = 1,050) and informant report (N = 1,200) standardization samples, as well as test-retest (n = 50 for self, n = 44 for informant) and inter-rater (n = 180) samples. Univariate BRs of elevated T scores were low (self-report = 3.3%-15.4%, informant report = 4.5%-16.3%). Multivariate BRs revealed the common occurrence of obtaining at least one elevated T-score across scales (self-report = 26.5%-37.3%, informant report = 22.7%-30.3%), whereas virtually none had elevated scores on all scales. Test-retest scores were highly correlated (self = .82-.94; informant = .91-.96). Inter-rater correlations ranged from .44 to .68. Significant (p < .05) test-retest T-score differences ranged from 7 to 12 for self-report, from 6 to 8 for informant report, and from 16 to 21 points for inter-rater T-score differences. Applications of these findings are discussed.


Bradycardia-tachycardia syndrome due to sinus node disease and concealed Wolff-Parkinson-White syndrome.
A 75-year-old woman was admitted to the hospital because of a supraventricular tachycardia, long sinus pauses and ventricular tachycardia of the torsades de pointe type. Temporary ventricular pacing suppressed ventricular arrhythmias but supraventricular tachycardia persisted. An electrophysiologic study revealed the coexistence of sick sinus syndrome and circus movement tachycardia due to a concealed left-sided accessory atrioventricular pathway. This case represents a previously undescribed form of the bradycardia-tachycardia syndrome.


Knee function and prevalence of osteoarthritis after isolated anterior cruciate ligament reconstruction using bone-patellar tendon-bone graft: long-term follow-up.
The aim of this investigation was to study patient-reported long-term clinical outcome, instrumental stablitity and prevalence of radiological osteoarthritis (OA) a minimum of ten years after isolated anterior cruciate ligament (ACL) reconstruction. An average of 13.5 years after ACL reconstruction with bone-patellar tendon-bone (BTB) autograft, 73 patients were evaluated. Inclusion criteria consisted of an isolated ACL rupture and reconstruction with BPTB graft with no associated intra-articular lesions, in particular, cartilage alterations or meniscal lesions. Clinical assessment was performed using the International Knee Documentation Committee (IKDC) and Tegner and Lysholm scores. Instrumental anterior laxity testing was carried out with the KT-1000™ arthrometer. Degree of degenerative changes and prevalence of OA were determined using the Kellgren- Lawrence scale. Mean follow-up was 13.5 years. Mean age was 43.8 years. About 75% of patients were graded A or B according to the IKDC score. The Lysholm score was 90.2 ± 4.8. Radiological assessment reported degenerative changes of grade II OA in 54.2% of patients. Prevalence of grades III or IV OA was found in 20%. The incidence of OA was significantly correlated with stability and function at long-term follow-up. Arthroscopic ACL reconstruction using BPTB autograft resulted in a high degree of patient satisfaction and good clinical results on long-term follow-up. A higher degree of OA developed in 20% of patients and was significantly correlated with increased anterior laxity at long-term follow-up.


Recurrence of post burn contractures of the elbow and shoulder joints: experience from a Ugandan hospital.
Recurrence of post-burn contractures, following inadequate management of post-burn contractures (PBC), is under reported. It is associated with multiple operations and an increased cost to patients and their families. The purpose of this study was to determine the frequency of recurrence of PBC of the shoulder and the elbow joint three months after surgical intervention and the associated risk- factors. This was a prospective cohort study conducted at CoRSU hospital from March 2012 to November 2014. All patients with PBC of the elbow and/or shoulder joint who consented to be in the study and met the inclusion criteria were enrolled. Data was collected using a pretested, coded questionnaire. A goniometer was used to measure the active range of motion of the involved joint. The measurements were recorded in degrees. The data was analysed with STATA version 12.1. 58 patients were enrolled consecutively in the study. There were 36 females and 22 males, with a female to male ratio of 1.6:1. The age range was 0.75-45 years, with a median age of 5 years. The average age at the time of injury was 3.4 years. The most common cause of initial burn injury was scalding. The average number of joints involved per patient was two. There was a high incidence of recurrence of PBC (52 %) among the participants. The shoulder had the highest frequency of recurrence at 67 %. The elbow joint had a frequency of recurrence of 27 %. All participants with both elbow and shoulder joint involvement had PBC recur. The risk factors for recurrence were flame burn (p = 0.007), duration of PBC of more than 1 year (p = 0.018), and incomplete release of the contracture (p = 0.002). The presence of keloids, hypertrophic scars, ulcers and the occurrence of complications at the contracture site were not associated with recurrence of PBC. Recurrence of PBC of the elbow and shoulder joint is a common problem. The risk factors should be kept in mind during management of PBC to reduce the recurrence rate.


Electron beam patterning of biomolecules.
The patterning of biomolecules on semiconducting surfaces is of central importance in the fabrication of novel biodevices. In the process of patterning, it is required that the biomolecule preserves its properties and the substrate is not damaged by the chemicals, the temperatures or the patterning beams involved in the procedure. Recently, both DUV and electron beam microlithography have been used in order to deposit protein layers in predefined patterns. Various approaches have been used, some involving photoresists. Contrast between exposed and unexposed regions, resolution of adjacent features and sensitivity to dose variation, are the key issues. The approach followed in this paper consists of a direct patterning of a biotin layer, deposited on an amino-silane primed silicon nitride surface, using an electron beam. After irradiation, the surface is covered by bovine serum albumin (BSA), which acts as a blocking material to protect the exposed areas from streptavidin adsorption. Using 20 keV e-beam energy and doses, in the range 100-1000 microC/cm(2), submicrometer dense lines of 1-microm pitch have been obtained. The results have been tested by fluorescence optical microscopy.


Ependymal cyst in a cynomolgus macaque (Macaca fascicularis).
A focal hypointense intracranial lesion was detected by magnetic resonance imaging in the right caudal occipital lobe of the cerebrum in an asymptomatic cynomolgus macaque (Macaca fascicularis). Following euthanasia, gross evaluation of the lesion revealed a 1 cm diameter, wedge-shaped intracranial cavitation without apparent communication with the ventricles. Histologically, the lesion was lined by ciliated cuboidal to low columnar epithelium that showed immunopositivity for cytokeratin, S-100, and GFAP. Based upon the gross, histologic, and immunohistochemical findings, this lesion was classified as an ependymal cyst. In the human patients, ependymal cysts are benign, non-infectious, non-inflammatory lesions of the central nervous system that are believed to originate from aberrant migration of ependymal cells during development. To our knowledge, this is the first report of an ependymal cyst in a non-rodent veterinary species and the first report of a benign intracranial cyst of any classification in a non-human primate.


Enthesitis may be one of the signs of severe disease in familial Mediterranean fever.
Familial Mediterranean fever (FMF) is an auto-inflammatory disease that is also characterized with some of the common musculoskeletal features of spondyloarthritis (SpA). Enthesitis is the hallmark of SpA. Recently, it was postulated that exertional leg pain is a possible sign of lower extremity enthesitis associated with FMF severity. In this study, we have evaluated the association between the enthesitis, enthesitis score and disease severity in FMF patients. We enrolled 238 FMF patients that fulfilled the modified Tel-Hashomer criteria. We assessed the presence of enthesitis at the Maastricht Ankylosing Spondylitis Enthesitis Score (MASES) defined sites with standard palpation method. Then, FMF patients dichotomised two groups as enthesitis group and controls. Herein, we evaluated the enthesis extensity with MASES. FMF disease severity was determined via the international severity scoring system for FMF (ISSF). Firstly, we have compared demographic properties, disease-related features and ISSF scores of the groups. Then, we have correlated ISSF with MASES in enthesitis group. We showed that 54 (22.6%) of 238 patients had enthesitis. The demographic features were similar between the groups. The enthesitis group had higher ISSF scores (p < 0.001); higher frequency of fever (p = 0.004), exertional leg pain (p < 0.001), myalgia (p < 0.001) and arthritis (p = 0.01); and more intense, widespread, frequent and longer attacks compared with controls. Moreover, there was a weak correlation between ISSF and MASES in the patients with enthesitis. Enthesitis may be a sign of more severe FMF phenotype and frequently associated with other musculoskeletal manifestations resemble SpA. Key points •More than one-fifth of the patients with FMF would suffer from enthesitis. •The FMF patients with enthesitis had higher ISSF scores; higher frequency of fever, exertional leg pain, myalgia and arthritis; and more intense, widespread, frequent and longer attacks as compared with controls. •Enthesitis may be a sign of more severe FMF phenotype and frequently associated with other SpA-like musculoskeletal feature.


The steroidal aromatase inhibitor exemestane prevents bone loss in ovariectomized rats.
The irreversible steroidal aromatase inhibitor exemestane (EXE) is one of three third generation aromatase inhibitors currently prescribed for advanced breast cancer in postmenopausal women. Its principal mechanism of action is to reduce estrogen by inhibiting its synthesis. In addition to its efficacy against breast cancer, its effects on other organs are important, especially when given to women with good-prognosis breast cancer or potentially to healthy women at increased risk of developing breast cancer. The purpose of this study was to evaluate the effects of EXE on bone and lipid metabolism in ovariectomized (OVX) rats. Ten-month-old Sprague-Dawley female rats were sorted into intact controls, intact + EXE, OVX controls, and OVX + EXE groups, and treated by weekly intramuscular injection with vehicle or 100 mg/kg EXE for 16 weeks. The bone mineral density (BMD), mechanical testing, histomorphometry, bone resorption marker-serum pyridinoline (PYD), and bone formation marker-serum osteocalcin (OC) were used to determine the effects of treatment on bone. In addition, total serum cholesterol, triglyceride, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were determined. BMD of the lumbar spine and femur were 11% and 7%, respectively, higher in OVX animals given EXE than in OVX controls (all Ps<0.001). Significant increases in the bending strength and toughness of the femora as well as the compressive strength and elastic modulus of the vertebrae were observed in OVX rats given EXE (all Ps<0.02 vs. OVX controls). Trabecular bone volume (BV) was significantly higher in OVX rats treated with EXE than in OVX controls (P<0.0001). In OVX animals, EXE reduced the OVX-induced increase of serum PYD by 96% (P<0.0001), and the OVX-induced increase of serum OC was completely prevented by treatment with EXE. In OVX animals, EXE resulted in a 28% reduction of serum cholesterol (P<0.0001) and reduced LDL by 64% compared with OVX controls (P<0.0001). The positive results of EXE on bone and lipid metabolism in the OVX rat model merit further investigation of the effects of EXE in postmenopausal women.


Using evolutionary constraint to define novel candidate driver genes in medulloblastoma.
Current knowledge of cancer genomics remains biased against noncoding mutations. To systematically search for regulatory noncoding mutations, we assessed mutations in conserved positions in the genome under the assumption that these are more likely to be functional than mutations in positions with low conservation. To this end, we use whole-genome sequencing data from the International Cancer Genome Consortium and combined it with evolutionary constraint inferred from 240 mammals, to identify genes enriched in noncoding constraint mutations (NCCMs), mutations likely to be regulatory in nature. We compare medulloblastoma (MB), which is malignant, to pilocytic astrocytoma (PA), a primarily benign tumor, and find highly different NCCM frequencies between the two, in agreement with the fact that malignant cancers tend to have more mutations. In PA, a high NCCM frequency only affects the BRAF locus, which is the most commonly mutated gene in PA. In contrast, in MB, >500 genes have high levels of NCCMs. Intriguingly, several loci with NCCMs in MB are associated with different ages of onset, such as the HOXB cluster in young MB patients. In adult patients, NCCMs occurred in, e.g., the WASF-2/AHDC1/FGR locus. One of these NCCMs led to increased expression of the SRC kinase FGR and augmented responsiveness of MB cells to dasatinib, a SRC kinase inhibitor. Our analysis thus points to different molecular pathways in different patient groups. These newly identified putative candidate driver mutations may aid in patient stratification in MB and could be valuable for future selection of personalized treatment options.


Evaluation of models for the evolution of protein sequences and functions under structural constraint.
In the field of evolutionary structural genomics, methods are needed to evaluate why genomes evolved to contain the fold distributions that are observed. In order to study the effects of population dynamics in the evolved genomes we need fast and accurate evolutionary models which can analyze the effects of selection, drift and fixation of a protein sequence in a population that are grounded by physical parameters governing the folding and binding properties of the sequence. In this study, various knowledge-based, force field, and statistical methods for protein folding have been evaluated with four different folds: SH2 domains, SH3 domains, Globin-like, and Flavodoxin-like, to evaluate the speed and accuracy of the energy functions. Similarly, knowledge-based and force field methods have been used to predict ligand binding specificity in SH2 domain. To demonstrate the applicability of these methods, the dynamics of evolution of new binding capabilities by an SH2 domain is demonstrated.


Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer.
A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII-MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.


The MDM2 inhibitor Nutlin-3 attenuates streptozotocin-induced diabetes mellitus and increases serum level of IL-12p40.
Besides its well-established oncosuppressor activity, a key function of p53 in regulating metabolic pathways has been recently identified. Nevertheless, the role of p53 with respect to diabetes mellitus (DM) appears highly controversial. To address this issue, we have used the cis-imidazoline compound Nutlin-3, an inhibitor of MDM2/p53 interaction, which represents a potent and selective non-genotoxic activator of the p53 pathway both in in vivo and in vitro experimental settings. Experimental DM was induced by intraperitoneal injections of low concentrations of streptozotocin (STZ) in C57BL/6N mice (n = 20). A group of control vehicle-injected mice (n = 10) and of STZ-treated mice (n = 10) was co-injected with Nutlin-3. Mice co-injected with STZ + Nutlin-3 exhibited attenuated features of DM with respect to animals treated with STZ alone. Indeed, STZ + Nutlin-3-treated mice were characterized by significantly (p < 0.05) lower levels of hyperglycemia, reduced weight loss, and increased spleen weight. In addition, STZ alone promoted a marked decrease in the levels of several circulating cytokines, including interleukin-12 (IL-12)p40. On the other hand, co-injection of STZ + Nutlin-3 significantly (p < 0.01) counteracted IL-12p40 down-modulation. In vitro experiments performed on the RAW264.7 macrophagic cell line model, used as cellular source of IL-12p40, demonstrated that Nutlin-3 treatment increased IL-12p40 release, strongly suggesting a direct effect of Nutlin-3 on the immune system. Overall, these data demonstrate that systemic administration of Nutlin-3 ameliorates the severity of STZ-induced DM and increases the levels of circulating IL-12p40.


Gender-specific transcriptional profiling of marine medaka (Oryzias melastigma) liver upon BDE-47 exposure.
Marine medaka (Oryzias melastigma) were exposed to 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) to investigate the gender-specific transcriptional profiles of liver tissue in response to this flame retardant. A cDNA library of O. melastigma was constructed, and 2304 clones were amplified from the library to fabricate a cDNA microarray. Sequences of these genes were assembled into 1800 sequences using Geneious, a bioinformatics software. Corresponding expressed sequence tags were blasted against the National Centre for Biotechnology Information non-redundant database and further classified into various biological categories according to the Gene Ontology project. Male and female three-month-old were fed a diet of BDE-47 contaminated Artemia at low dosage (290.3±172.3ng BDE-47/day) and high dosage (580.5±344.6ng BDE-47/day) for 5 and 21 days, respectively. The transcriptional profiles of O. melastigma liver were then generated by the species-specific cDNA microrarray. The results from microarray analysis suggested very different gene expression patterns between males and females for both BDE-47 exposure-dose and exposure-time, with male livers having stronger gene regulatory responses than female livers. Importantly, our results revealed that in male O. melastigma only, BDE-47 exposure may activate phosphoinositide-3-kinase and mitogen-activated protein kinase, proteins that play importance roles in cell growth, proliferation and survival.


Effects of diethyldithiocarbamate and fusaric acid upon memory storage processes in rats.
Diethyldithiocarbamate (DDC) and fusaric acid (FA), administered immediately posttraining, disrupted retention performance of rats trained on a one-trial inhibitory avoidance task. Posttraining DDC disrupted retention performance if the treatment was delayed up to at least 24 h (but not 72 h) after training. A single DDC injection also disrupted retention performance if the treatment was administered up to 10 days prior to training. The effects of DDC and FA upon several neurobiological measures were also investigated. Amnestic doses of DDC influence brain and adrenal catecholamine levels, brain electrographic activity and hippocampal Timm's staining. On the other hand, amnestic doses of FA only affected catecholamine levels. The amnestic effects of DDC were dissociated from brain seizure activity produced by DDC. The effects of DDC and FA upon retention may have been related to the drugs' effects upon catecholamine systems, but the bleaching by DDC of Timm's staining could reflect additional processes involved in DDC's effects upon retention. As opposed to the more widespread effects of DDC, FA's relative lack of effect upon other neurobiological measures more strongly supports a catecholaminergic interpretation for its effect upon retention.


Association of chlorphentermine with phospholipids in rat alveolar lavage materials, alveolar macrophages and type II cells.
Administration of chlorphentermine to rats leads to an increase in the phospholipid content of pulmonary surfactant materials and alveolar macrophages. It is known that this drug binds to pure phospholipids and prevents their degradation by phospholipases. Therefore, experiments were carried out to determine if chlorphentermine binds to surfactant phospholipids in vitro and to measure the in vivo association of drug with phospholipids in alveolar lavage materials from rats injected with [14C]chlorphentermine. The presence of chlorphentermine in alveolar macrophages, type II cells and other small pneumocytes (a population of lung cells which does not include alveolar macrophages or type II cells) from treated animals was also assessed. Binding of the drug to surfactant phospholipids, as measured with the fluorescent probe, 1-anilino-8-naphthalene sulfonate, occurs in vitro and does not differ in various subfractions of alveolar lavage materials isolated by differential centrifugation. Following daily administration of chlorphentermine to rats for 3 days, the drug appears to be associated with surfactant phospholipids such that the molar ratio is 1:100 (chlorphentermine/phospholipid). Chlorphentermine is also associated with alveolar macrophages (molar ratio, 1:18) and type II cells (molar ratio, 1:33). Not much drug is associated with the population of other lung cells (molar ratio, 1:333). In alveolar macrophages, approx. 70% of the drug seems to be bound to phospholipid and/or sequestered in subcellular organelles. However, only 20% of the chlorphentermine is bound and/or sequestered in type II cells. The results of these experiments suggest that following chlorphentermine administration, the drug is associated with phospholipids in acellular pulmonary lavage materials, alveolar macrophages and type II cells. This drug-phospholipid interaction may impair phospholipid degradation and lead to a phospholipidosis in surfactant materials and alveolar macrophages.


Adipocyte/macrophage fatty acid binding proteins control integrated metabolic responses in obesity and diabetes.
Fatty acid binding proteins (FABPs) are cytosolic fatty acid chaperones whose biological role and mechanisms of action are not well understood. Here, we developed mice with targeted mutations in two related adipocyte FABPs, aP2 and mal1, to resolve their role in systemic lipid, glucose, and energy metabolism. Mice lacking aP2 and mal1 exhibited a striking phenotype with strong protection from diet-induced obesity, insulin resistance, type 2 diabetes, and fatty liver disease. These mice have altered cellular and systemic lipid transport and composition, leading to enhanced insulin receptor signaling, enhanced muscle AMP-activated kinase (AMP-K) activity, and dramatically reduced liver stearoyl-CoA desaturase-1 (SCD-1) activity underlying their phenotype. Taken together with the previously reported strong protection against atherosclerosis, these results demonstrate that adipocyte/macrophage FABPs have a robust impact on multiple components of metabolic syndrome, integrating metabolic and inflammatory responses in mice and constituting a powerful target for the treatment of these diseases.


New Insights of Turmeric Extract-Loaded PLGA Nanoparticles: Development, Characterization and In Vitro Evaluation of Antioxidant Activity.
Here, we presented new insights of the development of poly(lactic-co-glycolic acid) nanoparticles containing turmeric compounds (turmeric-PLGA-NPs) using emulsion-solvent evaporation method. The nanoparticulate system was characterized by size, zeta potential, morphology, release profile, partition parameter, stability and encapsulation efficiency (%EE). Antioxidant activity studies were also evaluated. The Korsmeyer-Peppas model (Mt/M∞ vs. t) was used to determine the release mechanisms of the studied system. Our results demonstrated the emulsion-solvent evaporation method was shown advantageous for producing turmeric-PLGA-NPs in the range of 145 nm with high homogeneity in size distribution, zeta potential of -21.8 mV and %EE about 72%. Nanoparticles were stable over a period of one month. In vitro study showed a release of curcumin governed by diffusion and relaxation of the polymeric matrix. The partition parameter of the extract in relation to blank-PLGA-NPs was 0.111 ± 0.008 M-1, indicating a low affinity of curcumin for the polymer matrix. Antioxidant ability of the turmeric-PLGA-NPs in scavenging the radical 2,2-azinobis (3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) was inferior to free turmeric extract and showed a concentration and time-dependent profile. The study concluded that PLGA nanoparticles are potential carriers for turmeric extract delivery.


Cutaneous secondary follicular centre cell lymphoma in association with lymphoedema praecox.
We report a patient who developed cutaneous metastases in a lymphoedematous leg, from a follicular centre cell lymphoma, in a setting of lymphoedema praecox and recurrent deep-vein thromboses. To our knowledge, this is the first report of such a case. We discuss the possible causes for this localization.