Sustaining Knowledge Translation Practices: A Critical Interpretive Synthesis.
The health policy and systems research literature increasingly observes that knowledge translation (KT) practices are difficult to sustain. An important issue is that it remains unclear what sustainability of KT practices means and how it can be improved. The aim of this study was thus to identify and explain those processes, activities, and efforts in the literature that facilitate the sustaining of KT practices in health policy-making processes. We used a critical interpretive synthesis (CIS) to review the health policy and systems research and Science and Technology Studies (STS) literature. The STS literature was included as to enrich the review with constructivist social scientific perspectives on sustainability and KT. The CIS methodology allowed for creating new theory by critically combining both literatures. We searched the literature by using PubMed, Google Scholar, Web of Science, and qualitative sampling. Searches were guided by pre-set eligibility criteria and all entries were iteratively analysed using thematic synthesis. Eighty documents were included. Our synthesis suggests a shift from sustainability as an end-goal towards sustaining as actors' relatively mundane work aimed at making and keeping KT practices productive. This 'sustaining work' is an interplay of three processes: (i) translating, (ii) contexting, and (iii) institutionalising. Translating refers to activities aimed at constructing and extending networks. Contexting emphasises the activities needed to create contexts that support KT practices. Institutionalising addresses how actors create, maintain, and disrupt institutions with the aim of sustaining KT practices. The 'sustaining work' perspective of our CIS emphasises KT actors' ongoing work directed at sustaining KT practices. We suggest that this perspective can guide empirical study of sustaining work and that these empirical insights, combined with this CIS, can inform training programmes for KT actors, and thereby improve the sustainability of KT practices.


Electroresistance effect in ferroelectric tunnel junctions with symmetric electrodes.
Understanding the effects that govern electronic transport in ferroelectric tunnel junctions (FTJs) is of vital importance to improve the efficiency of devices such as ferroelectric memories with nondestructive readout. However, our current knowledge (typically based on simple semiempirical models or first-principles calculations restricted to the limit of zero bias) remains partial, which may hinder the development of more efficient systems. For example, nowadays it is commonly believed that the tunnel electroresistance (TER) effect exploited in such devices mandatorily requires, to be sizable, the use of two different electrodes, with related potential drawbacks concerning retention time, switching, and polarization imprint. In contrast, here we demonstrate at the first-principles level that large TER values of about 200% can be achieved under finite bias in a prototypical FTJ with symmetric electrodes. Our atomistic approach allows us to quantify the contribution of different microscopic mechanisms to the electroresistance, revealing the dominant role of the inverse piezoelectric response of the ferroelectric. On the basis of our analysis, we provide a critical discussion of the semiempirical models traditionally used to describe FTJs.


Cellular and in vivo electrophysiological effects of dronedarone in normal and postmyocardial infarcted rats.
We studied the effects of dronedarone (SR 33589) on the action potentials, membrane ionic currents, and arrhythmic activity in control rats and in rats after myocardial infarction, a model known to develop anomalous electrical activity. Dronedarone increased action potential duration in normal hearts. It had little effect on the action potentials that were already prolonged in the postmyocardial infarcted (PMI) rats. Particularly, dronedarone reduced the late sustained K(+) current, I(K) (or Isus) by 69%. Dronedarone induced only a tonic block of I(K). Similar relative inhibitions of I(K) by dronedarone were obtained in young, sham, and PMI rats, even if I(K) was less in sham than in young and further reduced in PMI rats. The EC(50) values were 0.78 and 0.85 microM in sham and PMI rats. Dronedarone induced a weak increase in the fast transient outward current, I(to). Time-to-peak and inactivation time constant of I(to) were decreased by dronedarone that also induced a marked slowing of I(to) recovery from inactivation. Similar effects were observed on the reduced I(to) recorded in PMI rats. Holter monitoring study in control, unthetered animals showed that dronedarone had no proarrhythmic effect. On rats, which after myocardial infarction exhibited ventricular premature beats, dronedarone significantly decreased beat occurrence during the 7-day treatment; this effect was sustained for two more weeks. Thus, dronedarone exerts antiarrhythmic effects on PMI rat heart. Its effects are attributable for the most part to the inhibition of outward K(+) currents and the increase in effective refractory period.


Synaptic effects of IL-1β and CRF in the central amygdala after protracted alcohol abstinence in male rhesus macaques.
A major barrier to remission from an alcohol use disorder (AUD) is the continued risk of relapse during abstinence. Assessing the neuroadaptations after chronic alcohol and repeated abstinence is important to identify mechanisms that may contribute to relapse. In this study, we used a rhesus macaque model of long-term alcohol use and repeated abstinence, providing a platform to extend mechanistic findings from rodents to primates. The central amygdala (CeA) displays elevated GABA release following chronic alcohol in rodents and in abstinent male macaques, highlighting this neuroadaptation as a conserved mechanism that may underlie excessive alcohol consumption. Here, we determined circulating interleukin-1β (IL-1β) levels, CeA transcriptomic changes, and the effects of IL-1β and corticotropin releasing factor (CRF) signaling on CeA GABA transmission in male controls and abstinent drinkers. While no significant differences in peripheral IL-1β or the CeA transcriptome were observed, pathway analysis identified several canonical immune-related pathways. We addressed this potential dysregulation of CeA immune signaling in abstient drinkers with an electrophysiological approach. We found that IL-1β decreased CeA GABA release in controls while abstinent drinkers were less sensitive to IL-1β's effects, suggesting adaptations in the neuromodulatory role of IL-1β. In contrast, CRF enhanced CeA GABA release similarly in controls and abstinent drinkers, consistent with rodent studies. Notably, CeA CRF expression was inversely correlated with intoxication, suggesting that CRF levels during abstinence may predict future intoxication. Together, our findings highlight conserved and divergent actions of chronic alcohol on neuroimmune and stress signaling on CeA GABA transmission across rodents and macaques.


Profile analysis: the effects of rove on sparse spectra.
Profile-analysis thresholds were measured in the presence and absence of overall level variation at different stimulus levels to determine whether nonlinear changes in the shape of the edges of excitation pattern peaks contribute to poorer spectral-shape sensitivity observed under roving levels. Roving levels decreased sensitivity for stimuli having few components separated widely in frequency to a greater extent than for stimuli having more densely spaced components. The stimulus level did not influence sensitivity when overall level variation was absent, suggesting that listeners rely on peaks in the excitation patterns and not the edges of the peaks (as would have been predicted by the near miss to Weber's law). Because the edges of the peaks were not used in the absence of roving levels, it follows that the larger rove effects for sparse stimuli were not likely due to excitation pattern inconstancy.


Keyhole limpet hemocyanin induces the activation and maturation of human dendritic cells through the involvement of mannose receptor.
Keyhole limpet hemocyanin (KLH) is a xenoantigen largely used in vitro as an immunogen to study primary antigen-specific T cell responses and in vivo as a vaccine component with optimal carrier qualities. So far, the mechanisms by which KLH exerts its immunostimulatory properties are still largely unknown. In particular, although dendritic cells (DCs) play a central role in the initiation and activation of immune responses, the effects of KLH on these cells have been poorly explored. In the present study we investigated the effects of KLH on DCs differentiated in vitro from human monocytes. We observed that KLH promotes the activation and maturation of DCs, as assessed by up-regulation of the surface expression of CD80, CD86, CD40, HLA-DR and CD83. Moreover, even if KLH stimulated the production of IL-12 and IL-10 by DCs, the final balance was clearly in favour of IL-12. According to these stimulatory effects, KLH significantly increased the allostimulatory activity of DCs. To verify whether these effects of KLH may be related to the binding of this highly glycosilated molecule to mannose receptor (MR), we performed inhibition experiments with anti-MR antibody. Results showed that the stimulatory activity of KLH is indeed partially mediated by its interaction with MR. Taken together, our results seem to indicate that KLH does promote the maturation of DCs endowed with the ability to stimulate cell-mediated immune responses. We suggest that this property of KLH may represent a novel further mechanism by which this molecule may exert its efficacy when co-administered with others antigens in immunotherapeutic protocols.


Aftershocks in a frictional earthquake model.
Inspired by spring-block models, we elaborate a "minimal" physical model of earthquakes which reproduces two main empirical seismological laws, the Gutenberg-Richter law and the Omori aftershock law. Our point is to demonstrate that the simultaneous incorporation of aging of contacts in the sliding interface and of elasticity of the sliding plates constitutes the minimal ingredients to account for both laws within the same frictional model.


Transmission electron microscopy studies of Moraxella (Branhamella) catarrhalis.
A trypsin-sensitive 200-kDa protein has been reported to be exclusively associated with haemagglutinating isolates of Moraxella (Branhamella) catarrhalis. Transmission electron microscopy studies revealed that haemagglutination by M. catarrhalis to both human and rabbit erythrocytes was mediated by a trypsin-sensitive outer fibrillar coat. This fibrillar layer was absent on non-haemagglutinating isolates examined. Immuno-electron microscopy, using a polyclonal antiserum containing antibodies to the 200-kDa protein as a probe, showed that the 200-kDa protein is present on the outer fibrillar layer of the bacterium. These findings suggest that the haemagglutinin of M. catarrhalis is a 200-kDa protein present on the outer fibrillar coat.


Sensitivity and specificity of quantitative difference SPECT analysis in seizure localization.
True ictal SPECT can accurately demonstrate perfusion increases in the epileptogenic area but often requires dedicated personnel waiting at the bedside to accomplish the injection. We investigated the value of perfusion changes as measured by ictal or immediate postictal SPECT in localizing the epileptogenic region in refractory partial epilepsy. Quantitative perfusion difference images were calculated by registering, normalizing and subtracting ictal (or immediate postictal) from interictal SPECT for 53 patients with refractory epilepsy. Perfusion difference SPECT results were compared with visually interpreted SPECT, scalp electroencephalography (EEG), MRI, PET and intracranial EEG. In 43 patients (81%), discrete areas of increased perfusion (with ictal injections) or decreased perfusion (with postictal injections) were noted. Interictal scalp EEG was localizing in 28 patients (53%), ictal scalp EEG was localizing in 35 patients (66%) and intracranial EEG was localizing in 22 patients (85%) (of 26 patients who underwent invasive study). MRI was localizing in 34 patients (64%), PET was localizing in 32 of 45 patients (71%), interictal SPECT was localizing in 26 patients (49%) and peri-ictal SPECT (visual interpretation) was localizing in 30 patients (57%). By comparison with an intracranial EEG standard of localization, SPECT subtraction analysis had 86% sensitivity and 75% specificity. Our data provide evidence that SPECT perfusion difference analysis has higher sensitivity and specificity than any other noninvasive localizing criterion and can localize epileptogenic regions with accuracy comparable with that of intracranial EEG. To obtain these results, one must apply knowledge of the timing of the ictal injection relative to seizure occurrence.


Necroptotic TNFα-Syndecan 4-TNFα Vicious Cycle as a Therapeutic Target for Preventing Temporomandibular Joint Osteoarthritis.
Temporomandibular joint osteoarthritis (TMJOA) is a chronic degenerative disease for which the underlying mechanism still remains unclear. Compared with apoptosis and autophagy, necroptosis causes greater harm to tissue homeostasis by releasing damage-associated molecular patterns (DAMPs). However, the role of necroptosis and downstream key DAMPs in TMJOA is unknown. Here, rodent models of TMJOA were established by the unilateral anterior crossbite (UAC). Transmission electron microscopy (TEM) and immunohistochemistry of receptor interacting protein kinase 3 (RIPK3)/phosphorylation of mixed lineage kinase domain-like protein (pMLKL) were conducted to evaluate the occurrence of necroptosis in vivo. The therapeutic effects of blocking necroptosis were achieved by intra-articularly injecting RIPK3 or MLKL inhibitors and using RIPK3 or MLKL knockout mice. In vitro necroptosis of condylar chondrocyte was induced by combination of tumor necrosis factor alpha (TNFα), second mitochondria-derived activator of caspases (SMAC) mimetics and carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (z-VAD-fmk). The possible DAMPs released by necroptotic chondrocytes were screened by quantitative proteomics and blocked by specific antibody. Translucent cytosol, swollen organelles, and ruptured cell membranes, features of necroptosis, were frequently manifested in chondrocytes at the early stage of condylar cartilage degeneration in TMJOA, which was accompanied by upregulation of RIPK3/pMLKL. Inhibiting or knocking out RIPK3/MLKL significantly prevented cartilage degeneration. DAMPs released by necroptotic condylar chondrocytes, such as syndecan 4 (SDC4) and heat shock protein 90 (HSP90), were verified. Furthermore, blocking the function of SDC4 significantly attenuated the expression of TNFα in cartilage and synovium, and accordingly increased cartilage thickness and reduced synovial inflammation. Thus, the necroptotic vicious cycle of TNFα-SDC4-TNFα contributes to cartilage degeneration and synovitis, and can serve as a potential therapeutic target for treating TMJOA. © 2022 American Society for Bone and Mineral Research (ASBMR).


1-Bromo-2-[(E)-2-nitro-ethen-yl]benzene.
In the title compound, C(8)H(6)BrNO(2), the dihedral angle between the planes of the benzene ring and the nitro group is 22.99 (12)°. In the crystal, inversion dimers associated by pairs of short Br⋯O contacts [3.2319 (17) Å] occur.


Impact of a pneumococcal conjugate vaccination program on carriage among children in Norway.
In July 2006, the seven-valent pneumococcal conjugate vaccine (PCV7) was introduced in Norway with a reduced (2 doses + 1 boost) dose schedule. Post-PCV7 shifts in pneumococcal reservoirs were assessed by two point prevalence studies of nasopharyngeal colonization among children in day care centers, before (2006) and after (2008) widespread use of PCV7. Nasopharyngeal swabs were obtained from 1,213 children, 611 in 2006 and 602 in 2008. A total of 1,102 pneumococcal isolates were recovered. Serotyping, multilocus sequence typing, and antimicrobial drug susceptibility testing were performed on all isolates. Although carriage of PCV7 serotypes decreased among both vaccinated and unvaccinated children, the overall prevalence of pneumococcal carriage remained high (80.4%) after vaccine introduction. The pneumococcal populations were diverse, and in the shift toward non-PCV7 serotypes, expansion of a limited number of established clonal complexes was observed. While non-antimicrobial-susceptible clones persisted among PCV7 serotypes, antimicrobial resistance did not increase among non-PCV7 serotypes. Direct and indirect protection of PCV7 against nasopharyngeal colonization was inferred from an overall decrease in carriage of PCV7 serotypes. No preference was found for nonsusceptible clones among the replacing non-PCV7 serotypes.


The use of synthetic oligonucleotides as protein inhibitors and anticode drugs in cancer therapy: accomplishments and limitations.
The function of gene products can be altered at many levels, including the mutation of gene sequence and the change in steady state levels of mRNA and/or protein by various mechanisms. The cumulative malfunction of specific gene products underlies many pathological conditions such as the multi-step and multi-cause acquisition of cancer. Here we discuss two oligonucleotide-based strategies in which these compounds target defective gene products acting either as antiprotein or anticode agents. The SELEX technique (systematic evolution of ligands by exponential enrichment) is an antiprotein approach in which nuclease-resistant DNA or RNA aptamers are selected by their ability to bind their protein targets with high affinity and specificity of the same range as antibodies. Such inhibitors were previously evolved against a great variety of targets, including receptors, growth factors and adhesion molecules implicated in the genesis of some kinds of cancer. Moreover, some results have already been obtained in animal models. The antigene technology interferes with earlier steps in the information flow leading from gene to protein. In this approach selective gene silencing is provided by the formation of stable and specific complexes between triplex forming molecules and their DNA targets. The feasibility of this strategy as well as a molecular mechanism for the action of antigene oligonucleotides has been demonstrated in cellular systems and in vivo. The use of oligonucleotide drugs (of either the antiprotein or the anticode type) as a viable approach to cancer therapy is limited by some common problems that will be discussed.


TSG-6 expression in human articular chondrocytes. Possible implications in joint inflammation and cartilage degradation.
The hyaluronan-binding protein TSG-6 (tumor necrosis factor-stimulated gene 6) forms a stable complex with the serine protease inhibitor, inter-alpha-inhibitor, potentiates the inhibition of plasmin activity, and has antiinflammatory effects in vivo. This study examines the expression of TSG-6 in human articular chondrocytes and cartilage. Human articular chondrocytes and cartilage explants were stimulated with cytokines, growth factors, and other agents. TSG-6 expression was analyzed by imaging-assisted Northern and Western blotting. TSG-6 messenger RNA (mRNA) expression was upregulated by cytokines and growth factors, predominantly interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha), platelet-derived growth factor AA (PDGF-AA), and transforming growth factor beta 1 (TGF beta 1). TSG-6 mRNA induction by TGF beta 1 was delayed as compared with IL-1beta. Treatment of the cells with the glucocorticoid dexamethasone neither induced TSG-6 mRNA nor did it affect IL-1 beta-induced transcript levels. TSG-6 mRNA induction may involve several signal transduction pathways. The strong transcriptional stimulation by phorbol myristate acetate suggests protein kinase C (PKC)-mediated signaling. In contrast, PKA- and Ca- dependent signals are only marginally involved as messengers leading to increased TSG-6 levels after IL-1beta and TNF alpha treatment. In chondrocyte and cartilage organ cultures, both free TSG-6 (35 kd) and the complex with inter-alpha-inhibitor (120 kd) were present and upregulated by IL-1 beta, TNF alpha, or TGF beta 1. Chondrocytes are a source of TSG-6 which may play a role in cartilage remodeling and joint inflammation.


The Marijuana Check-up: reaching users who are ambivalent about change.
A brief intervention called the Marijuana Check-up (MCU) was designed to attract adult marijuana users who were experiencing adverse consequences, but who were ambivalent about change and would be unlikely to seek treatment. Our objective was to determine whether the MCU would reach the target population. Comparisons were made between those who enrolled in the MCU versus those who were screened but failed to follow through with enrollment on demographic, drug use and stage of change variables. Comparisons were also made between participants in the MCU and participants in a concurrently offered treatment project that targeted marijuana users who wanted to quit. The study took place at the University of Washington in Seattle. Participants were adult marijuana users who telephoned and expressed interest in the MCU (n = 587). Study variables included stage of change, frequency and duration of drug use, DSM-IV cannabis dependence and abuse diagnoses and negative consequences of marijuana use assessed via interviews and questionnaires. Callers to the MCU were near-daily marijuana users, two-thirds of whom were in the pre-contemplation or contemplation stage of change. Participants who enrolled in the MCU reported fewer problems related to marijuana use and less readiness to make changes compared to those enrolled in the treatment study, despite similar levels of drug use. The MCU attracted and enrolled near-daily users of marijuana who experienced negative consequences but were ambivalent about making changes. The MCU potentially has a role in the continuum of care for substance abuse problems.


What remains of non-syndromic bicoronal synostosis?
More and more genetic syndromes are associated with bicoronal synostosis (BCS), making non-syndromic BCS (NSBCS) a shrinking entity. However, the numerical importance and clinical impact of syndromic BCS (SBCS) versus NSBCS have not been much studied. We retrospectively reviewed our experience with BCS over the last four decades in order to compare prevalence trends in SBCS and NSBCS. 195 patients were treated for BCS during the period 1978-2017: 104 (53.3%) were syndromic, 24 (12.3%) showed clinical and/or familial features suggesting a syndrome, although without final diagnostic confirmation, and 7 (3.5%) had associated extra-cranial malformations suggesting a syndromic context without identified genetic mutation; the remaining 61 (31.3%) were purely NSBCS. Surgery was required earlier in SBCS (21.7months, 95%CI 18.4-25.1) than in NSBCS (29.5months 95%CI 26.4-32.7). Prevalence of hydrocephalus and tonsillar herniation was significantly lower in NSBCS, and mortality concerned only SBCS. Prevalence of NSBC decreased significantly over the study period, likely because of more accurate testing, and decreased slightly over the last decade, possibly because of prenatal testing and abortion. NSBCS is now much less common than SBCS, and has a less aggressive clinical course, with lower rates of hydrocephalus, tonsillar herniation and mortality. This subgroup also deserves attention because it is likely that new discoveries are still to be made.


Purification and characterization of two monomeric kinesin constructs.
Steady-state and pre-steady-state kinetic methods were used to analyze two shorter Drosophila kinesin constructs (K341 and K366) in comparison to K401. K341, K366, and K401 represent the kinesin motor domains containing the N-terminal 341, 366, or 401 amino acids, respectively. K401 is dimeric (Kd = 37 +/- 17 nM) whereas both K366 and K341 are monomeric [Correia et al. (1995) Biochemistry 34, 4898-4907]. Like native kinesin and K401, K341 and K366 demonstrate low ATPase activity in the absence of microtubules (0.03 and 0.01 s-1, respectively), and ADP release is rate-limiting during steady-state turnover. Microtubules activate the steady-state ATPase to 84 s-1 for K341 (K(m),ATP = 100 microM; K0.5,MT = 3.2 microM tubulin) and 64 s-1 for K366 (K(m),ATP = 65 microM; K0.5,MT = 2.5 microM tubulin) in comparison to K401 at 20 s-1 (K(m)ATP = 60 microM; K0.5,MT = 1 microM tubulin). The rapid quench experiments for all three constructs show a burst of product formation during the first turnover, indicating the rate-limiting step for the microtubule-activated ATPase occurs after ATP hydrolysis. The interaction of K341 and K366 with the microtubule was analyzed by electron microscopy. The results show that K341 and K366, like K401, bind to the microtubule with an 8 nm axial periodicity. However, the addition of K366 to microtubules resulted in significant aggregation of microtubules. The pre-steady-state kinetic results show that K341 retains the kinetic and structural properties necessary to compare directly the kinetic properties of monomeric and dimeric kinesins, although the microtubule-activated ATPase is significantly faster for the monomeric constructs, suggesting possible interactions in the dimer which inhibit ATP turnover as part of the coupling to force production.


The impact of PET on the management of lung cancer: the referring physician's perspective.
(18)F-FDG PET is a molecular whole-body imaging modality that is increasingly being used for diagnosing, staging, and restaging cancer. The objective of this study was to determine referring physicians' perspectives on the impact of (18)F-FDG PET on staging and management of lung cancer. A questionnaire was sent to the 292 referring physicians of 744 consecutive patients with known or suspected lung cancer who were evaluated with PET. Questionnaires on 274 patients were returned (response rate, 37%). Management changes were categorized as intermodality (e.g., surgery to medical, surgery to radiation, and medical to no treatment) or intramodality (e.g., altered medical, surgical, or radiotherapy approach). The primary reasons for PET referral were staging of lung cancer in 61% of patients, diagnosis in 20%, and monitoring of therapy or the course of disease in 6%. Physicians reported that PET caused them to change their decision on clinical stage in 44% of all patients: The disease was upstaged in 29% and downstaged in 15%. PET resulted in intermodality management changes in 39% of patients, whereas 15% had an intramodality change. This survey-based study of referring physicians suggests that PET has a major impact on staging and management of lung cancer.


Rapid decline of fitness in panmictic populations of Drosophila melanogaster maintained under relaxed natural selection.
The parameters of the spontaneous deleterious mutation process remain poorly known, despite their importance. Here, we report the results of a mutation accumulation experiment performed on panmictic populations of Drosophila melanogaster without any genetic manipulations. Two experimental populations were kept for 30 generations under relaxed natural selection. Each generation, 100 pairs were formed randomly, and every fecund pair contributed a son and a daughter to the next generation. Comparison with two controls, one cryopreserved and the other kept as the experimental populations but with long generation time, showed that the number of surviving offspring per female declined by 0.2% and 2.0% per generation under benign and harsh, competitive conditions, respectively. Thus, the mutational pressure on fitness may be strong and depends critically on the conditions under which fitness is assayed.


Postoperative pain and contracture following total knee arthroplasty comparing parapatellar and subvastus approaches.
The purpose of this study was to see if subvastus approach would decrease incidence of postoperative contracture and pain following TKA compared to standard parapatellar approach. Retrospective review of 546 patients in Group A undergoing TKA using parapatellar approach were compared to 255 patients in Group B undergoing subvastus approach. No statistically significant differences regarding OR time, blood loss, BMI, or LOS. Total of 23 (4%) manipulations under anesthesia for contracture in Group A compared to 6 (2%) in Group B (p>0.05). Postoperative pain scores at 6 weeks was greater in Group A, p<0.05. We feel that a subvastus approach minimizes trauma to the extensor mechanism, and therefore decreases the incidence of postoperative pain following TKA.


Rapid naming ability in adults with stuttering.
The investigation of phonological encoding abilities in persons with stuttering has gained importance recently. There is limited information available on the rapid automatized naming ability in adults with stuttering. In the present study, rapid naming ability was assessed in 32 persons with stuttering and 32 persons without stuttering in the age range of 18-30 years on Rapid Automatized Naming and Rapid Alternating Stimulus test. The study findings indicated that the persons with stuttering took a longer time to complete the tasks as compared to persons without stuttering, suggestive of a possible breakdown in phonological encoding ability. Among the subtasks, the longer completion time was observed for the rapid alternating stimulus 3-set task and the color subtasks. The findings of the study clearly propose the need to encompass rapid naming skills, as a part of the assessment and management protocol for individuals with stuttering.


Methodology of pulsed photoacoustics and its application to probe photosystems and receptors.
We review recent advances in the methodology of pulsed time-resolved photoacoustics and its application to studies of photosynthetic reaction centers and membrane receptors such as the G protein-coupled receptor rhodopsin. The experimental parameters accessible to photoacoustics include molecular volume change and photoreaction enthalpy change. Light-driven volume change secondary to protein conformational changes or electrostriction is directly related to the photoreaction and thus can be a useful measurement of activity and function. The enthalpy changes of the photochemical reactions observed can be measured directly by photoacoustics. With the measurement of enthalpy change, the reaction entropy can also be calculated when free energy is known. Dissecting the free energy of a photoreaction into enthalpic and entropic components may provide critical information about photoactivation mechanisms of photosystems and photoreceptors. The potential limitations and future applications of time-resolved photoacoustics are also discussed.


The Cancer Clock Is (Not) Ticking: Links between Circadian Rhythms and Cancer.
Circadian rhythms regulate many physiological and behavioral processes, including sleep, metabolism and cell division, which have a 24-h oscillation pattern. Rhythmicity is generated by a transcriptional-translational feedback loop in individual cells, which are synchronized by the central pacemaker in the brain and external cues. Epidemiological and clinical studies indicate that disruption of these rhythms can increase both tumorigenesis and cancer progression. Environmental changes (shift work, jet lag, exposure to light at night), mutations in circadian regulating genes, and changes to clock gene expression are recognized forms of disruption and are associated with cancer risk and/or cancer progression. Experimental data in animals and cell cultures further supports the role of the cellular circadian clock in coordinating cell division and DNA repair, and disrupted cellular clocks accelerate cancer cell growth. This review will summarize studies linking circadian disruption to cancer biology and explore how such disruptions may be further altered by common characteristics of tumors including hypoxia and acidosis. We will highlight how circadian rhythms might be exploited for cancer drug development, including how delivery of current chemotherapies may be enhanced using chronotherapy. Understanding the role of circadian rhythms in carcinogenesis and tumor progression will enable us to better understand causes of cancer and how to treat them.


Association between serum folate with inflammatory markers, disease clinical activity and serum homocysteine in patients with inflammatory bowel disease. Does folate level have an effect on maintaining clinical remission?
Background Folate is an important vitamin with protective effect against some human diseases. The aim of this study was to evaluate the relationship between serum folate levels, inflammatory markers and disease clinical activity in patients with inflammatory bowel disease (IBD). Methods The participants were classified into two groups in which 38 IBD patients and 38 healthy controls were studied. Disease clinical activities were evaluated by means of established score systems. Serum folate, homocysteine and C-reactive protein and ESR were measured. Obtained data were analyzed with proper statistical methods and P- value less than 0.05 was considered as statistical significant. Results The level of serum folate was significantly reduced in IBD patients with active disease compared to patients with clinical remission (p=0.043) and also healthy controls (p = 0.008). Moreover, there was a significant inverse correlation between serum folate levels and C-reactive protein in IBD patients (r = -0.563 p =0.001). Conclusion Serum folate levels is associated with inflammatory markers and disease clinical activity in IBD patients, therefore there is a possibility that disease clinical activity is reduced with adequate folate level.


The Effect of Meal Glycemic Index and Meal Frequency on Glycemic Control and Variability in Female Nurses Working Night Shifts: A Two-Arm Randomized Cross-Over Trial.
Night shift workers are exposed to circadian disruption, which contributes to impaired glucose tolerance. Although fasting during the night shift improves glucose homeostasis, adhering to this dietary strategy may be challenging. This study evaluated the effect of fasting compared with the consumption of meals with different combinations of glycemic index (GI, low or high) and frequency (1 or 3 times) during the night shift on continuous glucose monitoring metrics. A 2-arm randomized cross-over trial was conducted on female nurses working night shifts. In each of those arms, the participants were either provided with no meal (fasted), low GI, or high-GI meal during the night shift with a meal frequency according to which arm they were randomly allocated to, either 1-MEAL or 3-MEAL. Outcome variables were glycemic control and variability (GC and GV) metrics during the night shift (21:30-7:00), in the morning after the night shift (07:00-13:00), and in the 24 h period (18:00-18:00). Compared to no meal, the consumption of 1 high-GI meal increased all GV metrics not only during the night shifts but also in the morning, for instance, as observed in the coefficient of variation (β = 0.03 mmol/L; 95% CI: 0.01, 0.05), and GV percentage (β = 4.13; 95% CI: 2.07, 6.18). The consumption of 1 or 3 low GI meals did not raise GC or GV metrics except for continuous overall net glycemic action during the night shifts after consuming 3 low GI meals. When controlling for GI, night shift meal frequency did not affect any metrics in any timeframe. High meal GI but not higher meal frequency during the night shift increased GC and GV in female night shift workers. Results for 1 low-GI meal during the night shift were not different from a glucose profile after no meal. This trial was registered at trialsearch.who.int as NL8715.


[Treatment of pediatric lithiasis in the lithotripsy and endourology unit of "Hospital La Luz". Commented retrospective of our case series].
The aim of the present article is to summarize the results we obtained treating children with urolithiasis over the last 30 years and to perform an analysis on the basis of the these results and relevant details according to national and international experience. Retrospective and descriptive statistical analysis of the 30 year experience in our clinics. The study was performed with a sample size of 178 children treated with urolithiasis that underwent 221 procedures. These procedures include ESWL, ureterorenoscopy (URS) and percutaneous nephrolithotomy (PCNL). We conclude in this study that ESWL in children was the most appropriate procedure for renal and proximal and middle-third ureteral lithiasis. Kidney stones measuring 2 to 3 cm can be treated without additional procedures or combined approaches. In contrast cystine stones caused the major problems for fragmentation. Moreover, the use of double J catheters increased the need for ESWL when catheter calcification occurred and endoscopic removal was impossible. The benefits of this method must be individually assessed both for the benefit of the temporary placement as well as for the expectation of permanence. We conclude that URS is the best choice for distal-third ureteral lithiasis and some cases of proximal and middle- third ureteral lithiasis. This enables for simultaneous treatments, ureteral dilatation and unexpected diagnoses. In particular, rigid ureteroscopy offers adjuvant options to ESWL with great therapeutic potential and easy handling. In consequence, to our good results and cost-benefit balance using ESWL and rigid URS (even combined), the use of flexible URS for renal lithiasis has been reduced. In general community units like ours, the combined PCNL has been indicated for particular complex cases, and open or laparoscopic surgery was not necessary in any case.


Changes in Levels of Intermediates of the C(4) Cycle and Reductive Pentose Phosphate Pathway under Various Concentrations of CO(2) in Maize Leaves.
The rate of CO(2) assimilation and levels of metabolites of the C(4) cycle and reductive pentose phosphate pathway in an attached leaf of maize (Zea mays L) were measured over a range of intercellular CO(2) concentration (Ci) of 10 to 190 microliters per liter. The CO(2) assimilation rate was saturated at a Ci of around 175 microliters per liter. The levels of ribulose 1,5-bisphosphate and fructose 1,6-bisphosphate decreased substantially with increasing Ci. The levels of 3-phosphoglycerate, phosphoenolpyruvate (PEP), and pyruvate increased with increasing Ci. The level of dihydroxyacetone phosphate increased moderately from Ci of 10 microliters per liter to 20 to 50 microliters per liter and stayed almost constant over the rest of the range of Ci investigated. The levels of fructose 6-phosphate did not show any significant changes over the range of Ci. The levels of glucose 6-phosphate decreased slightly with increasing Ci. Although photosynthetically inactive pools of malate, asparate, and alanine could mask real changes in levels of the photosynthetically active pools of these compounds, the apparent levels of these compounds and the total amount of intermediates in the C(4) cycle (malate, aspartate, pyruvate, PEP, and alanine) increased with increasing Ci. The results suggest that there is carbon input into the C(4) cycle from the reductive pentose phosphate pathway which increases the level of total intermediates of the C(4) cycle with increasing Ci.


Epidemiological study on the penicillin resistance of clinical Streptococcus pneumoniae isolates identified as the common sequence types.
There were some limitation in the current interpretation about the penicillin resistance mechanism of clinical Streptococcus pneumoniae isolates at the strain level. To explore the possibilities of studying the mechanism based on the sequence types (ST) of this bacteria, 488 isolates collected in Beijing from 1997-2014 and 88 isolates collected in Youyang County, Chongqing and Zhongjiang County, Sichuan in 2015 were analyzed by penicillin minimum inhibitory concentration (MIC) distribution and annual distribution. The results showed that the penicillin MICs of the all isolates covering by the given ST in Beijing have a defined range, either <0.25 mg/L or≥0.25 mg/L, except for the ST342. The isolates with penicillin MIC <0.25 mg/L were mainly collected before 2001, after which the isolates with MIC≥0.25 mg/L occurred and became the major population gradually. This law of year distribution, however, was not obvious for any specific ST. The isolates covering by any given ST could be determined with different penicillin MICs in the first few years after it was identified. The penicillin MIC of isolates identified as common STs and collected in Youyang County, Chongqing and Sichuan Zhongjiang County, including the ST271, ST320 and ST81, was around 0.25~2 mg/L (≥0.25 mg/L). Our study revealed the epidemiological distribution of penicillin MICs of the given STs determined in clinical S. pneumoniae isolates, suggesting that it is reasonable to research the penicillin resistance mechanism based on the STs of this bacteria.


Competitive Inhibition of the Enzyme-Mimic Activity of Gd-Based Nanorods toward Highly Specific Colorimetric Sensing of l-Cysteine.
Gd-based nanomaterials offer interesting magnetic properties and have been heavily investigated for magnetic resonance imaging. The applicability of these materials beyond biomedical imaging remains limited. The current study explores the applicability of these rare-earth nanomaterials as nanozyme-mediated catalysts for colorimetric sensing of l-cysteine, an amino acid of high biomedical relevance. We show a facile solution-based strategy to synthesize two Gd-based nanomaterials viz. Gd(OH)3 and Gd2O3 nanorods. We further establish the catalytic peroxidase-mimic nanozyme activity of these Gd(OH)3 and Gd2O3 nanorods. This catalytic activity was suppressed specifically in the presence of l-cysteine that allowed us to develop a colorimetric sensor to detect this biologically relevant molecule among various other contaminants. This suppression, which could either be caused due to catalyst poisoning or enzyme inhibition, prompted extensive investigation of the kinetics of this catalytic inhibition in the presence of cysteine. This revealed a competitive inhibition process, a mechanism akin to those observed in natural enzymes, bringing nanozymes a step closer to the biological systems.


The nuclear envelope, lamins and nuclear assembly.
The nuclear lamina is composed of both A- and B-type lamins and lamin-binding proteins. Many lamin-binding proteins are integral proteins of the inner nuclear membrane. Lamins and inner nuclear membrane proteins are important for a variety of cell functions, including nuclear assembly, replication, transcription, and nuclear integrity. Recent advances in the field in the past year include the identification of a family of spectrin-repeat-containing inner nuclear membrane proteins and other novel inner-membrane proteins, and the discovery of a nuclear membrane fusion complex. There is also growing evidence that A- and B-type lamins and their binding partners have distinct roles during nuclear assembly and interphase.


Protein S-mycothiolation functions as redox-switch and thiol protection mechanism in Corynebacterium glutamicum under hypochlorite stress.
Protein S-bacillithiolation was recently discovered as important thiol protection and redox-switch mechanism in response to hypochlorite stress in Firmicutes bacteria. Here we used transcriptomics to analyze the NaOCl stress response in the mycothiol (MSH)-producing Corynebacterium glutamicum. We further applied thiol-redox proteomics and mass spectrometry (MS) to identify protein S-mycothiolation. Transcriptomics revealed the strong upregulation of the disulfide stress σ(H) regulon by NaOCl stress in C. glutamicum, including genes for the anti sigma factor (rshA), the thioredoxin and MSH pathways (trxB1, trxC, cg1375, trxB, mshC, mca, mtr) that maintain the redox balance. We identified 25 S-mycothiolated proteins in NaOCl-treated cells by liquid chromatography-tandem mass spectrometry (LC-MS/MS), including 16 proteins that are reversibly oxidized by NaOCl in the thiol-redox proteome. The S-mycothiolome includes the methionine synthase (MetE), the maltodextrin phosphorylase (MalP), the myoinositol-1-phosphate synthase (Ino1), enzymes for the biosynthesis of nucleotides (GuaB1, GuaB2, PurL, NadC), and thiamine (ThiD), translation proteins (TufA, PheT, RpsF, RplM, RpsM, RpsC), and antioxidant enzymes (Tpx, Gpx, MsrA). We further show that S-mycothiolation of the thiol peroxidase (Tpx) affects its peroxiredoxin activity in vitro that can be restored by mycoredoxin1. LC-MS/MS analysis further identified 8 proteins with S-cysteinylations in the mshC mutant suggesting that cysteine can be used for S-thiolations in the absence of MSH. We identified widespread protein S-mycothiolations in the MSH-producing C. glutamicum and demonstrate that S-mycothiolation reversibly affects the peroxidase activity of Tpx. Interestingly, many targets are conserved S-thiolated across bacillithiol- and MSH-producing bacteria, which could become future drug targets in related pathogenic Gram-positives.


Carinal resection and reconstruction with complete pulmonary parenchyma preservation: a single-institution analysis of 36 cases.
Carinal resection, performed in only a few high-volume centers, remains one of the most complicated and technically demanding surgeries. Few studies have examined the outcomes of carinal resection and reconstruction with complete pulmonary parenchyma preservation. Patients who underwent isolated carinal resection and reconstruction at the Shanghai Chest Hospital between 2006 and 2020 were retrospectively reviewed. Clinicopathological, perioperative, and follow-up outcomes were analyzed. A total of 36 patients were included, including 19 men and 17 women. The average age was 50.7±14.8 years. Right posterolateral thoracotomy (n=33, 91.7%) and cross-field intubation during anastomosis (n=31, 86.1%) were selected for the majority of the carinal surgeries. The average intraoperative blood loss was 225.0 mL, and the mean operation duration was 196.1 minutes. Postoperative complications were observed in 14 patients (38.9%), including cicatricial stenosis (n=8, 22.2%), anastomotic fistula (n=3, 8.3%), air leak (n=1, 2.8%), cardiac arrhythmia (n=4, 11.1%), pneumonia (n=2, 5.6%), respiratory failure (n=1, 2.8%), and pulmonary embolism (n=1, 2.8%). There were 2 perioperative deaths (5.6%). Multivariate analysis revealed that being overweight was an independent favorable factor for postoperative complications [P=0.042, odds ratio (OR) =0.092, 95%, confidence interval (CI): 0.009-0.922]. Pathological diagnoses included squamous cell carcinoma (SCC) (n=12, 33.3%), adenoid cystic carcinoma (ACC) (n=15, 41.7%), mucoepidermoid carcinoma (MEC) (n=2, 5.6%), stricture (n=1, 2.8%), and other rare histological types. An R0 resection was achieved in 14 patients, while 21 patients (60.0%) had microscopically positive margins. Lymph node metastasis was confirmed in 6 patients (17.1%). Overall survival (OS) was 94.4% at 1 year and 79.4% at 5 years, with 107 months as the median survival time (95% CI: 64.0-150.0 months). All patients with negative margins remained alive during the follow-up period, while those who received R1 resections had much poorer survival rates due to tumor recurrence [P=0.042, hazard ratio (HR) =4.938, 95% CI: 1.062-22.950]. In selected patients, carinal resection and reconstruction with complete pulmonary parenchyma preservation was a feasible option to achieve an appreciable long-term survival at the risk of acceptable operative mortality and morbidity, particularly when complete resection with negative margins could be realized.


An interactive graphics system for real-time investigation and multivariate data portrayal for complex pedigree data systems.
GRIFFIN (graphics investigation of familial information), an interactive graphics system for exploratory investigation of data on individuals associated by familial relationships, was designed to provide genetic epidemiologists a flexible, rapidly responsive tool for viewing and guiding exploration of complex databases in the context of familiar pedigree structures. It graphically portrays both categorical and multivariate scalar data on individuals in those structures. The display can be inverted to show all ancestors and descendants of any individual the user designates. It provides cues to censored information when bushy pedigrees cannot be fully displayed without sacrificing legibility. These guide users on where to next invert the system. Investigators may translate/zoom the display, vary the mode of representing data, point to individuals to obtain displays of alphameric information about them, etc. Developed in Fortran using IBM's GDDM graphics subroutines for an IBM 3090 mainframe, GRIFFIN's design anticipates porting to smaller systems.


Reduce Driveline Trauma Through Stabilization and Exit Site Management: 30 Days Feasibility Results from the Multicenter RESIST Study.
The percutaneous lead management kit (PLMK) was developed for the HeartMate 2 (HM2) left ventricular assist device (LVAD) to reduce trauma at the exit site and to maintain a clean environment. REduce Driveline Trauma through StabIlization and Exit Site ManagemenT (RESIST) was a multicenter, prospective, nonrandomized study designed to evaluate the feasibility of the PLMK for managing the HM2 driveline exit site. Fifty patients were enrolled at five sites at a median of 495 days post-HM2 implant; 92% (46 of 50) of patients used the PLMK for a minimum of 30 days. At 30 days, more patients found the PLMK to be extremely comfortable (80% vs. 37%, p < 0.001) and extremely effective at stabilizing the driveline (82% vs. 40%, p < 0.001) compared with each center's standard of care. Frequency of dressing changes was 6-7 days or higher for 85% of the patients with PLMK. Three patients developed driveline infection while on PLMK (6%, 0.15 events per patient year), and 35 patients continued to use the PLMK after 6 months. The PLMK is easy to use, increases patient comfort, and increases driveline stability with a dressing change frequency of 6-7 days.


Lone motherhood and risk for cardiovascular disease: the National Population Health Survey (NPHS), 1998-99.
To examine whether lone parenthood among Canadian women is associated with risk factors for cardiovascular disease (CVD). The sample was from the National Population Health Survey (NPHS) and it included 2,184 mothers (women with dependent children < 18 years of age) of whom 478 were lone (22%) and 1,706 (78%) were partnered mothers. Descriptive statistics and multiple logistic regression were undertaken with weighted data. Compared to partnered mothers, lone mothers were significantly younger, and reported lower levels of education and income. Lone mothers reported significantly lower levels of health, less social support, and more distress and depression than partnered mothers. Further, lone mothers were significantly more likely to be smokers than partnered mothers. Odds ratios for being a lone mother and current smoker on social assistance in this analysis was 4.15 (95% CI 2.27-7.60), and for those reporting low or mid/high incomes respectively, were 2.99 (1.51-5.90) and 3.01 (1.93-4.70). There were no significant differences between the two groups for obesity or physical activity. Smoking is a significant CVD lifestyle risk for lone mothers. Smoking occurred in the context of considerable life stress, depression, and distress pointing to links between health behaviour, psychosocial factors, and CVD health. These results suggest that lone mothers are a key target group for primary prevention in the CVD field. Programs and policies that support lone mothers in their goals of attaining healthier lifestyles have the potential to contribute to the prevention of cardiovascular disease in Canadian women.


Technical note: The role of circulating low-density lipoprotein levels as a phenotypic marker for Holstein cholesterol deficiency in dairy cattle.
With the recent discovery of a Holstein cholesterol deficiency (HCD) haplotype, the USDA has labeled many dairy animals as HCD carriers based on haplotype and pedigree analysis. We set out to investigate the effect of HCD status on various cholesterol transport molecules, namely low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol, and triglycerides in both males and females. A genome-wide association study was also conducted to narrow down the genomic region correlated with varying LDL-C levels. In the study, 34 HCD carrier animals showed significantly lower cholesterol and LDL-C levels compared with their 34 closely related, non-HCD controls. The genome-wide association study based on 73 animals using 56,198 SNP markers revealed an association with chromosome 11 in the region of 66,218,925 to 66,946,746 bp. We also tested the effect of HCD status on sperm quality traits using fresh ejaculates and frozen-thawed semen samples, but did not find any discriminating effects. Our study has demonstrated the use of LDL-C as a key phenotypic marker for determining HCD status in dairy cattle and this is the first study that clearly shows a cause-effect relationship of the HCD haplotype on circulating LDL-C.


Two-dimensional ultrafast vibrational spectroscopy of azides in ionic liquids reveals solute-specific solvation.
The stereochemistry and the reaction rates of bimolecular nucleophilic substitution reactions involving azides in ionic liquids are governed by solute-solvent interactions. Two-dimensional ultrafast vibrational spectroscopy (2D-IR) shows that the picosecond dynamics of inorganic azides are substantially slower than organic azides in a series of homologous imidazolium ionic liquids. In water, both organic and inorganic azides spectrally diffuse with a ∼2 ps time constant. In the aprotic solvent tetrahydrofuran, both kinds of azides spectrally diffuse on a timescale >5 ps. In ionic liquids, like 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]), organic azides spectrally diffuse with a 2-4 ps time constant, and inorganic azides spectrally diffuse with a >40 ps time constant. Such a striking difference suggests that neutral (organic) and charged (inorganic) azides are incorporated in the ionic liquids with different solvation structures.


Tranexamic acid, an inhibitor of plasminogen activation, aggravates staphylococcal septic arthritis and sepsis.
Haemostatic balance shifts towards pro-coagulation during infection. Plasminogen, a key molecule of fibrinolysis, may play an important role in the pathogenesis of staphylococcal infections. In the present study, we assessed the impact of inhibition of plasminogen activation by tranexamic acid on the course of staphylococcal sepsis and septic arthritis in mice. We found significantly down-regulated plasmin activity and increased D-dimer levels in the blood from the mice with staphylococcal sepsis. Treatment with tranexamic acid significantly increased the severity and mortality of staphylococcal infection. In addition, tranexamic acid reduced the survival rate in a murine model for staphylococcal enterotoxin A-induced death. The aggravation of diseases by tranexamic acid was due neither to the pro-inflammatory cytokine network, nor to impairment of bacterial clearance. Modulation of fibrinolysis, either by supplement of fibrinolytic molecules (tissue plasminogen activator or plasmin) or by fibrinogen depletion, did not reduce the mortality of staphylococcal sepsis. In conclusion, we report that treatment with tranexamic acid led to distinct aggravation of staphylococcal septic arthritis and sepsis in mice, suggesting the clinical importance of fibrinolytic balance in staphylococcal infection.


Saturation mutagenesis of the E. coli RecA loop L2 homologous DNA pairing region reveals residues essential for recombination and recombinational repair.
The disordered mobile loop L2 of the Escherichia coli RecA protein is known to play a central role in DNA binding and pairing. To investigate the local chemical environment in relation to function we performed saturation mutagenesis of the loop L2 region (amino acid positions 193-212) using a site-directed mutagenesis procedure, and determined the recombinational proficiency of the 380 mutants using genetic assays for homologous recombination and recombinational repair. Residues Asn193, Gln194, Arg196, Glu207, Thr209, Gly211, and Gly212 were identified as stringently required for recombinational events in bacterial cells. In addition, our findings suggest the involvement of loop L2 in the ATPase activity of RecA, and a role for residues Gln194, Arg196, Lys198 and Thr209 in the DNA-dependent hydrolysis of ATP. Finally, since 20 residue peptides that comprise this region can pair homologous DNAs by forming filamentous beta-structures, we propose how the information from the mutant analysis might facilitate the use of a simplified amino acid alphabet to design beta-structure forming L2 peptides with improved RecA-like activities.


Novel animal models for studying complex brain disorders: BAC-driven miRNA-mediated in vivo silencing of gene expression.
In schizophrenia, glutamic acid decarboxylase 1 (GAD1) disturbances are robust, consistently observed, cell-type specific and represent a core feature of the disease. In addition, neuropeptide Y (NPY), which is a phenotypic marker of a sub-population of GAD1-containing interneurons, has shown reduced expression in the prefrontal cortex in subjects with schizophrenia, suggesting that dysfunction of the NPY+ cortical interneuronal sub-population might be a core feature of this devastating disorder. However, modeling gene expression disturbances in schizophrenia in a cell type-specific manner has been extremely challenging. To more closely mimic these molecular and cellular human post-mortem findings, we generated a transgenic mouse in which we downregulated GAD1 mRNA expression specifically in NPY+ neurons. This novel, cell type-specific in vivo system for reducing gene expression uses a bacterial artificial chromosome (BAC) containing the NPY promoter-enhancer elements, the reporter molecule (eGFP) and a modified intron containing a synthetic microRNA (miRNA) targeted to GAD1. The animals of isogenic strains are generated rapidly, providing a new tool for better understanding the molecular disturbances in the GABAergic system observed in complex neuropsychiatric disorders such as schizophrenia. In the future, because of the small size of the silencing miRNAs combined with our BAC strategy, this method may be modified to allow generation of mice with simultaneous silencing of multiple genes in the same cells with a single construct, and production of splice-variant-specific knockdown animals.


Palliation of malignant rectal obstruction from invasive prostate cancer with multiple overlapping self-expanding metal stents.
Self-expandable metal stents (SEMS) are used for colonic neoplastic and extracolonic metastatic obstruction relief. Limited data exists on their use for locally invasive prostate cancer. We describe a unique approach using overlapping SEMS to alleviate a rectosigmoid obstruction from locally invasive prostate cancer. A patient with locally advanced prostate cancer presented with obstipation and lymphedema. Placement of overlapping rectosigmoid SEMS was performed, relieving the visualized rectosigmoid obstruction.


[Intraoperative monitoring of intact parathyroid hormone during surgery for primary hyperparathyroidism].
The aim of the present study was to evaluate a new immunometric assay for intraoperative parathyroid hormone monitoring. The test was applied in 70 patients who underwent surgery for primary hyperthyroidism (pHPT) between 6/1999 and 6/2001. Among these patients, 61 showed a solitary adenoma, eight a hyperplasia and one a double adenoma. Intraoperative iPTH samples were taken at the beginning of the operation and 5, 10 and 15 min after removal of the parathyroid gland. Criterion for a successful operation were a decrease of iPTH levels of more than 50 % within 5 min and of more than 60 % within 15 min after parathyroidectomy. Following the removal of a solitary adenoma, iPTH levels decreased by 63 % (+/- 13 %) after 5 min and by 76 % (+/- 10 %) after 15 min respectively. In case of hyperplasia, a significant decrease of iPTH levels was not observed until a subtotal parathyroidectomy had been carried out. In the present study there were 2 false negative and one false positive results corresponding with a sensitivity of 97 % and a specificity of 89 % for prediction of a solitary adenoma. In our opinion, intraoperative iPTH monitoring using this new assay allows the safe distinction between adenoma and multiglandular disease. It represents a valuable adjunct to surgical skill as it permits minimally invasive operations for solitary adenomas, and in case of recurrent surgery helps to detect the region of interest by selective venous sampling for parathyroid hormone.


Scanning of key residues in antibody binding sites by two saturation-mutagenesis approaches.
The complementarity-determining region 3 of the heavy chain (CDRH3) generally contributes the most to antibody-antigen binding. His101H in CDRH3 of the antibody Se155-4, which is specific for a trisaccharide epitope of Salmonella serotype B O-antigen, was mutated systematically into all nineteen other amino acids by a double mutation approach. Enzyme immunoassay (EIA) and affinity chromatography showed that the Asn, Gln, Gly and Ser mutants exhibited moderate to strong activity. Some mutants, such as Thr and Pro, had weak binding activity, while the acidic and hydrophobic amino acid substitutions resulted in complete loss of activity. A second mutation approach which randomly changed a selected residue into all other nineteen amino acids, while precluding wild-type transformants, is also described.


Safety and biocompatibility of injectable vaccine adjuvants composed of thermogelling block copolymer gels.
Injectable thermogelling polymers have been recently investigated as novel adjuvants and delivery systems for next generation vaccines. As research into natural and synthetic biocompatible polymers progresses, the safety and biocompatibility of these compounds is of paramount importance. We have developed cationic pentablock copolymer (PBC) vaccine adjuvants based on Pluronic F127, a thermogelling triblock copolymer that has been approved by the FDA for multiple applications, and methacrylated poly(diethyl amino)ethyl methacrylate outer blocks. These novel materials have been demonstrated to effectively create an antigen depot, minimally impact antigen stability, and enhance the immune response to antigens (i.e., adjuvanticity) in mice. In this work, we investigated the safety and biocompatibility of the parent triblock Pluronic gels and the cationic PBC gels in mice. Histological analysis showed no injection site reactions and no damage to the liver or kidneys was observed upon administering the block copolymer formulations. However, the subcutaneous injection of a thermogelling Pluronic solution induced increased levels of lipids in the blood, with no further deleterious effects observed from the addition of the cationic outer blocks. This hyperlipidemia resolved within 30 days after the administration of the Pluronic formulation. To mitigate this adverse effect, the vaccine adjuvant formulations were modified by adding poly(vinyl alcohol), which allowed gelation, while reducing the amount of Pluronic in the formulation. This modified formulation abrogated the observed hyperlipidemia and no adverse effects were observed in the serum through biomarker analysis or at the injection site (i.e., inflammation) in comparison to the responses induced by administration of saline or incomplete Freund's adjuvant. These studies provide a foundation to developing these gels as adjuvants for next generation vaccines. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1754-1762, 2019.


Fourteen- vs seven-day bismuth-based quadruple therapy for second-line Helicobacter pylori eradication.
To compare the efficacy of 14- and 7-d bismuth-based quadruple therapies as second-line eradication treatment for Helicobacter pylori (H. pylori) infection. Between 2004 and 2014, the medical records of 790 patients who had experienced failure of first-line proton pump inhibitor (PPI)-based eradication therapy and were then treated with bismuth-based quadruple therapy were retrospectively reviewed. Those who received bismuth-based quadruple therapy [PPI, bismuth, metronidazole, and tetracycline (PBMT)] for either 7 d or 14 d were assigned to a PBMT-7 group (n = 543) or a PBMT-14 group (n = 247), respectively. The eradication rates for both groups were determined by intention-to-treat (ITT) and per-protocol (PP) analyses. ITT analysis compared the treatment groups as originally allocated while the PP analysis including only those patients who had completed the treatment as originally allocated. Successful eradication therapy for H. pylori infection was defined as a negative (13)C-urea breath test 4 wk after the end of eradication treatment. The overall ITT eradication rate was 69.1% (546/790). Final ITT eradication rates were 67.4% (366/543; 95%CI: 63.1%-71.7%) in the PBMT-7 group and 72.8% (180/247; 95%CI: 67.4%-78.2%) in the PBMT-14 group (P = 0.028). The overall PP eradication rate was 80.0% (546/682), and the final PP eradication rates were 78.2% (366/468; 95%CI: 72.1%-84.0%) in the PBMT-7 group and 84.1% (180/214; 95%CI: 76.8%-90.8%) in the PBMT-14 group (P = 0.009). The H. pylori eradication rates in the PBMT-14 group were significantly higher than in the PBMT-7 group according to both ITT (P = 0.028) and PP analysis (P = 0.009). Compliance was similar in both groups (PBMT-7 group: 97.9%; PBMT-14 group: 96.4%). Adverse event rates were 10.7% (51/478) and 17.1% (38/222) in the PBMT-7 and PBMT-14 groups, respectively (P = 0.487). The 14-d bismuth-based quadruple therapy is a significantly more effective second-line eradication treatment for H. pylori infection than the 7-d alternative.


Availability of 15N from pioneer herbaceous plants to pine seedlings in reclaimed burnt soils.
A pot experiment was used to assess N uptake by pine seedlings during 2 years on a burnt soil to which was added (15)N-labelled ryegrass, obtained from a (15)N-enriched sample of this soil after a fire. The nitrogen concentration in needles, stems and roots of seedlings decreased significantly from the first to the second growing period (from 2.55, 1.30 and 2.19% to 1.19, 0.47 and 1.00%, respectively), with needles accounting for 53-58% of the pine-N. At the end of the experiment, 98.87 +/- 1.12% of the added ryegrass-(15)N was recovered: two-thirds in the soil organic N pool and one-third in the pine seedlings. Therefore, the post-fire pulse of inorganic-N, which was successfully kept in the burnt soil-plant system through its uptake by the pioneer species, is available for trees in the medium term. Pine seedlings assimilated 16.4% and 16.9% of the added ryegrass-(15)N in the first and second year, respectively. This result contrasts with the usual yearly decrease of added N uptake by plants; a possible explanation is the transient increase of available N in burnt soils that would have modified the mineralization pattern of the (15)N-labelled phytomass. The pine-N derived from the ryegrass-N decreased from 4.05% in the first year to 2.53% in the second one, with 3.10% being the 2-year weighed average. In addition to the direct contribution of ryegrass to pine-N nutrition reflected by these figures, the rapid post-fire establishment of a herbaceous cover on the burnt soil also provides important indirect benefits for tree nutrition by reducing organic- and inorganic-N losses.


Another family with a euchromatic duplication variant of 9q13-q21.1 derived from segmentally duplicated pericentromeric euchromatin.
Microscopically visible copy number variations within the proximal short arm heterochromatin and proximal long arm of chromosome 9 have been described as euchromatic variants (EVs) and are derived from extensive segmental duplications (SDs) that map to both the proximal short and long arms of chromosome 9. Recently, 3-4 additional copies of an SD cassette were found in 2 families with duplication EVs of 9q13-q21. Here, we report a third family with a duplication EV of 9q13-q21.1 that was ascertained at prenatal diagnosis for advanced maternal age and found in the fetus and her phenotypically normal mother. Dual-colour fluorescence in situ hybridization with bacterial artificial chromosomes RP11-246P17 and RP11-211E19 was consistent with the EV chromosome having 1-2 additional copies of a similar SD cassette, except that the SD-boundary clone RP11-88I18 was not apparently included. It is important to distinguish the 9q13-q21.1 EVs from possible pathogenic imbalances of chromosome 9, especially at prenatal diagnosis, as these EVs have no established phenotypic or reproductive consequences. The nature of the G-dark bands in 9q13-q21 EVs is briefly discussed.


A model of temporary migration: the Egyptian case.
"In this article, a family migration model is introduced where differences in regional amenities and wage rates can result in voluntary, temporary migration. A household production model of temporary migration will be developed and tested to analyse Egyptian temporary migration to the Gulf region. The theoretical model will be a two-period model of temporary migration and will be empirically tested using the well known and comprehensive 1982 Egyptian Fertility Survey, which is part of the World Fertility Survey and covers a period for which temporary migration from Egypt was at or near its peak. This was also before the Gulf War, after which patterns of migration were different." (SUMMARY IN FRE AND SPA)


Rapid Detection of Dimethoate in Soybean Samples by Microfluidic Paper Chips Based on Oil-Soluble CdSe Quantum Dots.
Given the imperative of monitoring organophosphorus pesticides (OPs) residues in the ecosystem, here a novel, facile and sensitive fluorescence sensor is presented for the rapid detection of dimethoate. In this work, surface molecularly imprinted polymer (SMIP) and microfluidic technology had been introduced to enhance the selectivity and portability of the described methodology. Oil-soluble CdSe quantum dots (QDs) synthesized in a green way were used as fluorescent material for the selective detection of dimethoate on the basis of static quenching and photoinduced electron transfer mechanism. Among many kinds of paper materials, glass fiber paper was used as the novel substrate of paper chip due to low pristine fluorescence and better performance when combining CdSe QDs. In the process of molecular imprinting, the interaction between several functional monomers and dimethoate molecule was investigated and simulated theoretically by software to improve the selectivity of the sensor. Consequently, the fabricated novel detection platform could effectively respond to dimethoate in 10 min with the concentration range of 0.45-80 μmol/L and detection limit of 0.13 μmol/L. The recovery in the spiked experiment soybean sample was in an acceptable range (97.6-104.1%) and the accuracy was verified by gas chromatography-mass spectrometry, which signified the feasibility and potential in food sampling.


An improved approximation to the exact solution of the two-stage clonal growth model of cancer.
Multistage clonal growth models are of interest for cancer risk assessment because they can explicitly incorporate data on cell replication. Both approximate and exact formulations of the two stage growth model have been described. The exact solution considers the conditional probability of tumors arising in previously tumor-free animals; the approximate solution estimates total probability of tumor formation. The exact solution is much more computationally intensive when time-dependent cell growth parameters are included. The approximate solution deviates from the exact solution at high incidences and probabilities of tumor. This report describes a computationally tractable, 'improved approximation' to the exact solution. Our improved approximation includes a correction term to adjust the unconditional expectation of intermediate cells based on the time history of formation of intermediate cells by mutation of normal cells (recruitment) or by cell division in the intermediate cell population (expansion). The improved approximation provided a much better match to the exact solution than the approximate solution for a wide range of parameter values. The correction term also appears to provide insight into the biological factors that contribute to the variance of the expectation for the number of intermediate cells over time.


X-ray Moiré deflectometry using synthetic reference images.
Moiré fringe deflectometry with grating interferometers is a technique that enables refraction-based x-ray imaging using a single exposure of an object. To obtain the refraction image, the method requires a reference fringe pattern (without the object). Our study shows that, in order to avoid artifacts, the reference pattern must be exactly matched in phase with the object fringe pattern. In experiments, however, it is difficult to produce a perfectly matched reference pattern due to unavoidable interferometer drifts. We present a simple method to obtain matched reference patterns using a phase-scan procedure to generate synthetic Moiré images. The method will enable deflectometric diagnostics of transient phenomena such as laser-produced plasmas and could improve the sensitivity and accuracy of medical phase-contrast imaging.


Cerebral and cardiac amyloidosis in autopsied elderly individuals.
Amyloidosis in elderly individuals can be an independent alteration and a characteristic of aging. However, the clinical, pathophysiologic, and biochemical characteristics of amyloidosis related to age remain uncertain. The purpose of this study was to determine the extent to which the heart and/or the brain of individuals aged 60 years or over exhibits amyloid deposits. The autopsy findings of individuals who were at least 60 years old were studied. The autopsies took place between the years of 1976 and 2000. A total of 10 cases were selected that had hearts without cardiopathies, had negative serology for Chagas' disease, and had brains without morphological changes related to encephalopathies. Slides with fragments of heart and brain were processed and analyzed using polarized and common light microscopy. Of the 10 cases, 4 were positive for amyloidosis. All had positive findings in the brain, and 1 case also had positive findings in the heart. Among the positive cases, 50% were of people aged 60 to 69 years. There appeared to be a relationship between the presence of amyloid deposits and the ratio of brain and body weight, with the ratio in the positive cases being smaller than in the negative cases. The analysis of amyloid deposits in the brains and hearts of elderly individuals shows that such deposits may lead to a systemic attack of senility, common to natural aging. It is not certain that beta-amyloid deposits would alone bring such drastic repercussions to the individual. Some additional disorders of the organism could cause the breakdown of the natural balance related to the accumulation of these proteins, leading the way to the pathological contexts of amyloidosis.


Characterization of neuronal and vascular histamine receptors mediating the salivary and vasodilator responses to histamine of the dog submandibular gland.
The submandibular gland in situ was perfused with blood through the glandular artery at constant pressure in anesthetized dogs, and all drugs were administered intra-arterially. During infusion of metiamide, histamine and 2-(2-pyridyl)ethylamine (PEA) produced salivary and vasodilator responses consisting of an early and a late component. The dose-response curves for respective components of the salivary and vasodilator responses to PEA were parallel with the corresponding curves for histamine and in producing these responses PEA was about 40 times less potent than histamine on a molar basis. During infusion of mepyramine, histamine and dimaprit produced only the early vasodilator response. The dose-vasodilator response curves to histamine and dimaprit were parallel, and dimaprit was about 750 times less potent than histamine on a molar basis. The present results support the conclusion obtained in a previous study that neuronal histamine receptors mediating the whole salivary and the late vasodilator response are exclusively of the H1-type and vascular histamine receptors mediating the early vasodilator response consist of both H1-and H2-type although the former is predominant.


Development of therapeutic approaches to 'triple negative' phenotype breast cancer.
Triple negative phenotype (TNP) breast cancers are characterised by the lack of expression of oestrogen and progesterone receptors and of human EGF receptor 2 (HER2) overexpression/amplification. This subgroup of cancers has an aggressive clinical behaviour and is associated with poorer overall survival compared with other subtypes. Given the lack of targets for current tailored therapies in TNP tumours, chemotherapy is the only systemic treatment available; however, overall outcomes remain poor. Therefore, optimal treatment regimens and targeted therapies are urgently needed. We discuss characteristics of TNP cancers that underpin the rationale of current and novel therapeutic strategies, and an approach for finding and validating new therapeutic targets. The results of large prospective randomised controlled trials are currently awaited. Efforts to unravel the heterogeneity and complexity of TNP cancers using the latest high-throughput molecular techniques and integrating these findings with biology-driven therapeutic strategies in clinical trials will be of paramount importance for the development of treatment approaches for this breast cancer subtype.


Extracellular superoxide dismutase is upregulated with inducible nitric oxide synthase after NF-kappa B activation.
Inflammatory cytokines have been shown to upregulate secretion of the antioxidant enzyme extracellular superoxide dismutase (EC-SOD) in dermal fibroblasts and, in other cells, to stimulate production of nitric oxide (.NO). Because superoxide rapidly scavenges .NO, forming the injurious peroxynitrite anion (OONO-), we hypothesize that stimulated cells upregulate EC-SOD expression concurrently with .NO release. To test for coregulation of EC-SOD and .NO within the same cell, the timing of inducible nitric oxide synthase (iNOS) and EC-SOD transcription was measured after exposure of a rate type II pneumocyte analog, the L2 cell line, to a combination of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). Upregulation of iNOS and EC-SOD transcription occurred after 6 h of exposure, and transcription of both genes was linked by activation of the transcription factor nuclear factor-kappa B. Both EC-SOD and iNOS were elevated in rat lung homogenates 24 h after intratracheal instillation with IFN-gamma and TNF-alpha. The observation that EC-SOD and iNOS are temporally coregulated after cytokine exposure suggests the possibility of a critical mechanism by which cells might protect .NO and avoid the formation of OONO- during inflammation.


Chemical vapor deposition of highly adherent diamond coatings onto co-cemented tungsten carbides irradiated by high power diode laser.
The present investigation deals with the definition of a new eco-friendly alternative to pretreat Co-cemented tungsten carbide (WC-Co) substrates before diamond deposition by hot filament chemical vapor deposition (HFCVD). In particular, WC-5.8 wt %Co substrates were submitted to a thermal treatment by a continuous wave-high power diode laser to reduce surface Co concentration and promote the reconstruction of the WC grains. Laser pretreatments were performed both in N(2) and Ar atmosphere to prevent substrate oxidation. Diamond coatings were deposited onto the laser pretreated substrates by HFCVD. For comparative purpose, diamond coatings were also deposited on WC-5.8 wt %Co substrates chemically etched by the well-known two-step pretreatment employing Murakami's reagent and Caro's acid. Surface morphology, microstructure, and chemical composition of the WC-5.8 wt %Co substrates after the different pretreatments and the deposition of diamond coatings were assessed by surface profiler, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction analyses. Wear performance of the diamond coatings was checked by dry sliding linear reciprocating tribological tests. The worn volume of the diamond coatings deposited on the laser pretreated substrates was always found lower than the one measured on the chemically etched substrates, with the N(2) atmosphere being particularly promising.


Therapeutic effects of shaogan fuzi decoction in rheumatoid arthritis: Network pharmacology and experimental validation.
Shaogan Fuzi Decoction (SGFD), one of the classical prescriptions of Chinese Medicine, has a long history in the treatment of rheumatoid arthritis (RA), but definitive studies on its efficacy and mechanism of action are lacking. This study aims to elucidate the pharmacodynamic role of SGFD against RA and the potential mechanisms based on a combination of network pharmacology and experimental verification. The RA model in rats was induced by intradermal injection of bovine type Ⅱ collagen and incomplete Freund's adjuvant at the tail root. SGFD was administered once a day by oral gavage for 4 weeks. After SGFD administration, rat's arthritis index (AI) score and paw swelling decreased to some extent, and synovial inflammation, vascular hyperplasia, and cartilage destruction of the ankle joint were improved. Simultaneously, thymus and spleen index and serum levels of C-reactive protein (CRP) were lowered. Network pharmacology revealed that quercetin, kaempferol, naringenin, formononetin isorhamnetin and licochalcone A were the potentialiy active components, and IL6, TP53, TNF, PTGS2, MAPK3 and IL-1β were potential key targets for SGFD in the treatment of RA. Ingredients-targets molecular docking showed that the components had the high binding activity to these target proteins. The mechanism of SGFD for RA involves various biological functions and is closely correlated with TNF signaling pathway, Osteoclast differentiation, T cell receptor signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, NF-κB signaling pathway, toll-like receptor signaling pathway, and so on. Western blot and ELISA showed that the expression of toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB) p65, phosphorylated c-Jun N-terminal kinase (p-JNK), p-p38, phosphorylated extracellular regulated kinase (p-ERK) and TNF-α was significantly upregulated in the synovium of RA rats, and the levels of serum inflammatory factors were significantly increased. SGFD inhibits the activation of the TLR4/NF-κB/MAPK pathway and the expression/production of pro-inflammatory cytokines. In summary, SGFD could improve the symptoms and inflammatory response in collagen-induced arthritis (CIA) rat model. The mechanism might be related to the regulation of TLR4/MAPKs/NF-κB signaling pathway and the reduction of inflammatory factor release, which partially confirms the results predicted by network pharmacology.


Effects on different full-coverage designs and materials of crack propagation in first mandibular molar: an extended finite element method study.
Objectives: This study aims to investigate the biomechanical properties of fracture resistance in cracked teeth using five different full-coverage restorations made of three different materials. Materials and Methods: A 3D model of a mandibular first molar was created to design five different full-coverage repair models: crown, crown with composite resin filling inside, occlusal veneer, occlusal veneer with composite resin filling inside and onlay. These repair models were fabricated using three different materials, namely, zirconia, lithium disilicate (LDS), and a hybrid polymer-infiltrated ceramic network material (PIC). In total, 15 repair models were tested using the extended finite element method (XFEM), with an occlusal load of 5000 N applied slowly to the occlusal surface of the restoration. The analysis of stress distribution in the restoration and dentin crack line was conducted to measure and record the crack initial load on the restoration and dentin. Results: The results showed that restorations on the occlusal surface significantly improved crack resistance, with zirconia exhibiting superior fracture resistance among the materials tested. Restorations of crown with composite resin filling inside demonstrated the highest resistance to fracture, while occlusal veneers showed the lowest. MPS concentration was observed at the interface between the restoration and dentin and at the root bifurcation, with the highest values at the top of crack development. Dentin covered by oxidized restorations had the highest displacement, while PIC restorations exhibited the lowest. Pulp analysis revealed selective MPS concentration and strain patterns in models with zirconia restorations and onlay, with pronounced pulp displacement in zirconia restorations and onlay. Enamel analysis indicated larger MPS values and displacements in zirconia restoration models and onlay, with higher strain in onlay. Restoration played a crucial role in protecting the tooth, with crack propagation initial loads in dentin surpassing restorations in experimental groups. Conclusion: This study confirms that full-coverage restorations significantly increased the fracture resistance of cracked teeth, with zirconia restorations significantly protecting the underlying cracked tooth. Elimination of fracture lines in the restoration design can improve fracture resistance in cracked teeth. The findings have implications for dental prosthetic design and clinical practice.


The Japan Home-health Apparatus Industrial Association: investigation of home-use electronic sphygmomanometers.
The Japanese Home-health Apparatus Industrial Association is an official independent organization comprising ten departments. That concerned with home electronic sphygmomanometers, which has seven participants from different Japanese manufacturers, has already undertaken and is currently involved in various activities related to voluntary standards for performance validation and quality assurance. Because Japanese companies form a large proportion of manufacturers, these activities are important in terms of autonomic regulation. Although many improvements have been made to home electronic sphygmomanometers, some problems still remain unresolved, especially in terms of measurement reliability and easy operation by lay people. Another aspect of the department's work relates to making proposals on major validation standards, such as those of the Association for the Advancement of Medical Instrumentation, the British Hypertension Society and Comité Européen de Normalisation (CEN). Clinical validation should be discussed in order to define a more accurate standard method of measurement using auscultation and more appropriate criteria that are unaffected by primary blood pressure variation.


Sharp DNA bends as landmarks of protein-binding sites on straightened DNA.
We have developed a fluorescence-based method for mapping single or multiple protein-binding sites on straightened, large-size DNA molecules (> 5 kbp). In the described method, protein-DNA complexes were straightened and immobilized on a flat surface using surface tension. A fraction of the immobilized complexes displayed a sharp DNA bend with two DNA segments extending from the apex. The presence of DNA-binding proteins at the apex was verified by atomic force microscopy. The position of protein binding relative to the ends of the DNA molecule was determined by measuring the length of two DNA segments using fluorescence microscopy. We demonstrate the potential of the fluorescence-based method to localize protein-binding sites on the DNA template and to evaluate relative binding affinity. The proposed protein-binding-site mapping technique is simple and easy to perform. Practical applications include screening for DNA-binding proteins and the localization of protein-binding sites on large segments of DNA.


Transplant Prognosis in Kidney Transplant Recipients with Diabetes under Mycophenolic Acid-Focused Therapeutic Drug Monitoring.
Mycophenolate mofetil is a key immunosuppressant that is metabolized into mycophenolic acid (MPA). The prognostic impact of MPA-focused therapeutic drug monitoring on allograft prognosis has not been determined in kidney transplant recipients with diabetes. In this study, we assessed the pharmacokinetics of MPA and allograft prognosis in recipients with diabetes. This study retrospectively analyzed 64 adult kidney transplant recipients. MPA blood concentration data (e.g., the time to the maximum concentration (Tmax), and the area under the concentration-time curve from 0 to 12 h (AUC0-12)) were collected at 3 weeks and 3 months after kidney transplantation. Of the 64 recipients, 15 had pre-existing diabetes. At 3 months after kidney transplantation, the Tmax of MPA was significantly longer in recipients with diabetes (mean (standard deviation): 2.8 (2.1) h) than in recipients without diabetes (1.9 (1.1) h, p = 0.02). However, the allograft estimated glomerular filtration rate and acute rejection rate, including borderline change, did not differ according to the diabetes status in patients with adjusted AUC0-12 of MPA within the target range. In conclusion, a longer Tmax of MPA was observed in recipients with diabetes; however, acceptable allograft prognosis was observed in kidney transplant recipients with diabetes and a sufficient AUC0-12 of MPA.


Academy of medicine, Singapore-Ministry of Health clinical practice guidelines: management of food allergy.
The Academy of Medicine, Singapore (AMS) and the Ministry of Health (MOH) publish clinical practice guidelines to provide doctors and patients in Singapore with evidence-based guidance on managing important medical conditions. This article reproduces the introduction and executive summary (with recommendations from the guidelines) from the AMS-MOH clinical practice guidelines on the Management of Food Allergy, for the information of readers of the Singapore Medical Journal. Chapters and page numbers mentioned in the reproduced extract refer to the full text of the guidelines, which are available from the Academy of Medicine website: http://www.ams.edu.sg/guidelines.asp#foodallergy. The recommendations should be used with reference to the full text of the guidelines. Following this article are multiple choice questions based on the full text of the guidelines.


Platelet dysfunction during cardiopulmonary bypass assessed by a novel whole-blood aggregometer.
The purpose of this study was to assess perioperative platelet function with 2 types of monitors (a whole-blood aggregometer [WBA analyzer; Mebanix, Tokyo, Japan]) and the Sonoclot monitor [Sienco, Wheat Ridge, CO]) in patients undergoing hypothermic cardiopulmonary bypass (CPB). Prospective, observational study. Single-center study at a university hospital. Twenty-six patients who underwent coronary artery bypass grafting or valve replacement under hypothermic CPB without platelet transfusion or fresh frozen plasma administration. Blood sampling was performed at the following time periods: after anesthetic induction, after CPB, and on the first postoperative day. These samples were assessed with the WBA analyzer and the Sonoclot. Significant attenuation of adenosine diphosphate-induced platelet aggregation was detected shortly after CPB by 2 WBA analyzer-derived parameters: a decrease in the filtration pressure rate and an increase in the platelet aggregatory threshold index. Platelet aggregation returned to the preoperative level on the next day. There was no correlation between the amount of postoperative mediastinal drainage and defects in platelet aggregation. On the other hand, time to peak obtained by the Sonoclot did not show any significant changes. Whole-blood aggregation measured with the WBA analyzer detected transient platelet dysfunction shortly after CPB, whereas the Sonoclot was less sensitive to this change.


Measuring adriamycin-induced cardiac hemodynamic dysfunction with a proteomics approach.
Adriamycin is a potent antitumor drug that causes severe cardiotoxicity. However, the toxic mechanisms are not clear. We used a proteomics approach to analyze changes in protein profiles after adriamycin-induced changes in hemodynamic factors. Although adriamycin itself did not affect left ventricular developed pressure (LVDP) or left ventricular end diastolic pressure (LVEDP), the drug did enhance susceptibility to ischemia-reperfusion-induced changes in LVDP, LVEDP and heart rate. Adriamycin altered the expression of 52 proteins, primarily energy metabolism and cytoskeleton proteins. Adriamycin decreased the expression of the metabolism-related proteins, ATP synthase, Sdha protein, Triose phosphate isomerase 1 (TPI-1), pyruvate dehydrogenase E1 alpha1, 6-phosphofructokinase, and fructose-1,6-bisphosphatase, as did cytoskeletal proteins, such as actin. Alterations in energy metabolism and subsequent free radical production may affect cytoskeletal protein expression, producing adriamycin-induced changes in cardiac hemodynamics.


Orthopedic protraction of the maxilla may affect cranial base synchondroses indicated by increased expressions of growth factors.
To examine the biological adaptation of cranial base synchondroses (CBS) when the maxilla was forward positioned by orthopedic force. The Department of Orthodontics at Shanghai Jiao Tong University. 50 Sprague-Dawley rats, 4 weeks of age, were divided into experimental (n=30) and control groups (n=20). An orthopedic appliance was fitted to the cranio-maxillary complex to advance the maxilla forward. The animals in the experimental group, together with the counterparts in the control group, were sacrificed at days 1, 3, 5, 7, and 14, respectively. The whole cranial base housing both the spheno-ethmoid (SES) and spheno-occipital synchondroses (SOS) was removed for tissue processing and immunotest of Sox9, Core-binding factor α 1 (Cbfa1), and vascular endothelial growth factor (VEGF), three carefully selected growth factors that are markers of chondrogenesis in different stages and its transition to endochondral ossification. Semiquantitative analysis was also conducted by using a computerizing imaging system. The temporal tendency of the changes in the expressions of the three growth factors featured an increase from Day 3 and onwards for Cbfa1 and VEGF, and a following decline after Day 5 for Sox9. In both SES and SOS, the expressions of the three growth factors were significantly stronger in the experimental groups than that in groups (p<0.05). Protractive orthopedic force imposed on the maxilla provokes an enhancement of chondrogenic process in CBS.


The effect of hypothalamic catecholamine synthesis inhibition by alpha-methyltyrosine on gonadotropin secretion during sexual maturation in the male Wistar rat.
alpha-Methyl-p-tyrosine (AMPT), a competitive inhibitor of tyrosine hydroxylase, was used to block the synthesis of hypothalamic catecholamines in six different age groups (15, 23, 30, 38, 56, and 72 days of age) in both intact and castrated male rats. Castrated rats received testosterone-filled Silastic capsules of various lengths to give four graded serum concentrations of testosterone in each age group. Intact and castrate animals of all ages were killed 1 week after the castrate group was orchiectomized and outfitted with Silastic capsules. On the day of death, 250 mg/kg BW AMPT base were given ip at zero time, and a second dose (125 mg/kg BW) was given 2 h after the first injection. Rats were killed at zero time and 2 and 4 h after the first AMPT injection. Serum was collected for LH, FSH, and testosterone determinations. AMPT had marked biphasic effects on LH and T secretion, causing both to increase (less than 70% above basal) in the intact immature rats (15 and 23 days of age) and fall (greater than 60% below basal) in the mature rats (38, 56, and 72 days of age). LH increased (up to 307%) in the castrated immature rats in response to AMPT, but no change occurred in the mature castrated rats until moderate concentrations of testosterone were present (1000-2000 pg/ml). Then, LH, as in intact rats, decreased (up to 90%) during exposure to AMPT. These data suggest that the ratio of stimulatory and inhibitory effects of hypothalamic norepinephrine and dopamine changes during sexual maturation, with inhibitory effects predominating in the prepubertal rat.


Management of dementia-related behavioral disturbances: a nonpharmacologic approach.
To describe practical nonpharmacologic approaches to dementia-related behavioral problems for enhancement of the function and care of elderly patients with dementia. We reviewed the pertinent medical literature and summarized strategies and available resources for management of geriatric patients with dementia and behavior problems. For optimal care of older patients with dementia, key concepts of related behavior problems must be understood. Agitation and aggression, resisting help with care, wandering, incontinence, sleep disturbance, and emotional lability can become difficult management issues with such patients. In some patients, these disruptions can lead to overmedication and nursing home placement. Herein, practical nonpharmacologic measures for management of behavior problems that arise among community-dwelling and institutionalized elderly patients with dementia are discussed. Attention is directed to the medical, psychologic, environmental, and social factors that may contribute to unwanted behaviors. Nonpharmacologic approaches can help ameliorate behavioral problems and assist in the overall care of elderly patients with dementia.