The role of alpha-, beta-, and gamma-ENaC subunits in distal lung epithelial fluid absorption induced by pulmonary edema fluid.
Edema fluid (EF) increases epithelial Na(+) transport by rat fetal distal lung epithelia (FDLE) and induces net lung fluid absorption in fetal mouse lung explants [Rafii B, Gillie DJ, Sulowski C, Hannam V, Cheung T, Otulakowski G, Barker PM, O'Brodovich H. J Physiol (Lond) 544: 537-548, 2002]. We now show that EF increases fluid absorption across monolayers of rat FDLE in a dose-dependent manner. To study the role of subunits of the epithelial Na(+) channel (ENaC) in the phenomena, we cultured explants from the distal lungs of 16-day gestational age wild-type (WT) or alpha-, beta-, or gamma-ENaC knockout or heterozygote (HT) mice. WT explants cultured in media continuously expanded over time as a result of net fluid secretion. In contrast, when explants were exposed to EF for 24 h, net fluid absorption occurred. EF-exposed explants had normal histology, but marked changes were seen after Triton X-100 or staurosporine exposure. Transmission electron microscopy showed EF promoted lamellar body formation and abundant surfactant in the explants' lumens. EF-induced changes in explant size were similar in alpha-ENaC knockout, WT, and HT littermate fetal lung explants (P > 0.05). In contrast, EF's effect was attenuated in beta- and gamma-ENaC knockouts (P < 0.05) vs. WT and HT littermate fetal lung explants. EF exposure slightly decreased or had no effect on mRNA levels for alpha-ENaC in various mouse genotypes but decreased expression of beta- and gamma-ENaC subunit mRNAs (P < 0.01) across all genotype groups. We conclude that beta- and gamma-, but not alpha-, ENaC subunits are essential for EF to exert its maximal effect on net fluid absorption by distal lung epithelia.


Intranasal MMI-0100 Attenuates Aβ1-42- and LPS-Induced Neuroinflammation and Memory Impairments via the MK2 Signaling Pathway.
Background: Accumulating evidence suggests inhibiting neuroinflammation as a potential target in therapeutic or preventive strategies for Alzheimer's disease (AD). MAPK-activated protein kinase II (MK2), downstream kinase of p38 mitogen activated protein kinase (MAPK) p38 MAPK, was unveiled as a promising option for the treatment of AD. Increasing evidence points at MK2 as involved in neuroinflammatory responses. MMI-0100, a cell-penetrating peptide inhibitor of MK2, exhibits anti-inflammatory effects and is in current clinical trials for the treatment of pulmonary fibrosis. Therefore, it is important to understand the actions of MMI-0100 in neuroinflammation. Methods: The mouse memory function was evaluated using novel object recognition (NOR) and object location recognition (OLR) tasks. Brain hippocampus tissue samples were analyzed by quantitative PCR, Western blotting, and immunostaining. Near-infrared fluorescent and confocal microscopy experiments were used to detect the brain uptake and distribution after intranasal MMI-0100 application. Results: Central MMI-0100 was able to ameliorate the memory deficit induced by Aβ1-42 or LPS in novel object and location memory tasks. MMI-0100 suppressed LPS-induced activation of astrocytes and microglia, and dramatically decreased a series of pro-inflammatory cytokines such as TNF-α, IL-6, IL-1β, COX-2, and iNOS via inhibiting phosphorylation of MK2, but not ERK, JNK, and p38 in vivo and in vitro. Importantly, one of the reasons for the failure of macromolecular protein or peptide drugs in the treatment of AD is that they cannot cross the blood-brain barrier. Our data showed that intranasal administration of MMI-0100 significantly ameliorates the memory deficit induced by Aβ1-42 or LPS. Near-infrared fluorescent and confocal microscopy experiment results showed that a strong fluorescent signal, coming from mouse brains, was observed at 2 h after nasal applications of Cy7.5-MMI-0100. However, brains from control mice treated with saline or Cy7.5 alone displayed no significant signal. Conclusions: MMI-0100 attenuates Aβ1-42- and LPS-induced neuroinflammation and memory impairments via the MK2 signaling pathway. Meanwhile, these data suggest that the MMI-0100/MK2 system may provide a new potential target for treatment of AD.


Effect of different simulated altitudes on repeat-sprint performance in team-sport athletes.
This study aimed to assess the impact of 3 heights of simulated altitude exposure on repeat-sprint performance in team-sport athletes. Ten trained male team-sport athletes completed 3 sets of repeated sprints (9 × 4 s) on a nonmotorized treadmill at sea level and at simulated altitudes of 2000, 3000, and 4000 m. Participants completed 4 trials in a random order over 4 wk, with mean power output (MPO), peak power output (PPO), blood lactate concentration (Bla), and oxygen saturation (SaO2) recorded after each set. Each increase in simulated altitude corresponded with a significant decrease in SaO2. Total work across all sets was highest at sea level and correspondingly lower at each successive altitude (P < .05; sea level < 2000 m < 3000 m < 4000 m). In the first set, MPO was reduced only at 4000 m, but for subsequent sets, decreases in MPO were observed at all altitudes (P < .05; 2000 m < 3000 m < 4000 m). PPO was maintained in all sets except for set 3 at 4000 m (P < .05; vs sea level and 2000 m). BLa levels were highest at 4000 m and significantly greater (P < .05) than at sea level after all sets. These results suggest that "higher may not be better," as a simulated altitude of 4000 m may potentially blunt absolute training quality. Therefore, it is recommended that a moderate simulated altitude (2000-3000 m) be employed when implementing intermittent hypoxic repeat-sprint training for team-sport athletes.


On Johnson's (2000) Relative Weights Method for Assessing Variable Importance: A Reanalysis.
This article provides a reanalysis of J. W. Johnson's (2000) "relative weights" method for assessing variable importance in multiple regression. The primary conclusion of the reanalysis is that the derivation of the method is theoretically flawed and has no more validity than the discredited method of Green, Carroll, and DeSarbo (1978) on which it is based. By means of 2 examples, supplemented by other results from the literature, it is also shown that the method can result in materially distorted inferences when it is compared with another widely used importance metric, namely, general dominance (Azen & Budescu, 2003; Budescu, 1993). Our primary recommendation is that J. W. Johnson's (2000) relative weights method should no longer be used as a variable importance metric for multiple linear regression. In the final section of the article, 2 additional recommendations are made based on our analysis, examples, and discussion.


Skin Improvement with Antioxidant Effect of Yuja (Citrus junos) Peel Fractions: Wrinkles, Moisturizing, and Whitening.
Yuja (Citrus junos) has been cultivated and used for food and medicinal purposes in China and Korea. Its antioxidant, anti-wrinkle, moisturizing, and whitening effects were evaluated in HaCaT, HDF, and B16F10 cells. UVB has been known to cause cellular stress and the production of reactive oxygen species (ROS). Ambivalence of oxidative stress has been reported; however, excessive levels of ROS contribute to skin aging through the loss of elasticity and collagen fibers of connective tissue in the dermis. Skin aging is one of the biological processes that is affected by various factors, including UVB. Pro-Collagen I and hyaluronic acid contents were measured in UVB-irradiated HaCaT and HDF cells to evaluate the anti-wrinkle and moisturizing effects of Yuja-peel (YJP) fractions in -EA (ethyl acetate), -Hex (hexane), and -BuOH (butanol). The expression of matrix metalloproteinases (MMPs) involved in collagen degradation was confirmed to be inhibited by YJP fractions at both the protein and mRNA levels. Filaggrin and serine palmitoyltransferase (SPT), which are moisturizing factors, were induced by YJP fractions. B16F10 cells were treated with α-MSH to induce hyperpigmentation, and then the whitening efficacy of YJP fractions was verified by observing a decrease in melanin content. Overall, our results contribute to the development of various novel skin-improving cosmetics and pharmaceuticals with YJP fractions as active ingredients.


Complications associated with the Bernese periacetabular osteotomy for hip dysplasia in adolescents.
The Bernese (Ganz) periacetabular osteotomy is an effective surgical procedure to reorient the acetabulum, allowing restoration of anatomic femoral head coverage and medial translation of the hip in adults with hip dysplasia. However, it is a challenging surgical procedure, and we know of no study that has specifically analyzed the complications and associated factors seen with this procedure in adolescent patients. A retrospective clinical and radiographic review of a consecutive series of adolescent patients who underwent a Bernese periacetabular osteotomy for hip dysplasia was conducted. Eighty-three osteotomies were performed in seventy-six patients with an average age (and standard deviation) of 15.6 +/- 2.4 years. Significant improvement from the preoperative to the two-year follow-up evaluation was seen radiographically with regard to the lateral center-edge angle (-0.14 degrees to 35.5 degrees), the ventral center-edge angle (-5.13 degrees to 31.3 degrees), and the femoral head extrusion index (38.4% to 7.7%) (p < 0.0001 for all). There were three major complications, including excessive arterial bleeding requiring embolization in a patient with a prior acetabuloplasty, osteonecrosis of the acetabular fragment in a patient with severe dysplasia and subluxation of the hip, and osteonecrosis of the femoral head following combined periacetabular and femoral osteotomies in a patient with Charcot-Marie-Tooth disease. Eighteen hips (22%) had minor complications, including nonunion of the superior pubic ramus osteotomy (five hips), a superficial stitch abscess (four), and transient lateral femoral cutaneous nerve palsy (four). Nine hips (11%) underwent removal of symptomatic screws, and two required a second operation to reposition the acetabular fragment. An underlying diagnosis other than developmental dysplasia increased the prevalence of minor complications (p = 0.0017), while a major complication was more likely with longer surgery time, greater blood loss, and proximal femoral osteotomy. The Bernese periacetabular osteotomy is a joint-preserving procedure that very effectively corrects acetabular dysplasia in adolescent patients, providing improved radiographic results and a low rate of complications. Although the rate of minor complications is increased when there is an underlying diagnosis other than developmental dysplasia, no other predictors were identified. However, a major complication is more likely with a longer duration of surgery and with a concomitant femoral varus osteotomy.


A switch-like dynamic mechanism for the initiation of replicative senescence.
Telomeres are specialized structures protecting chromosomes against genome instability. Telomeres shorten with cell division, and replicative senescence is induced when telomeres are badly eroded. Whereas TRF2 (telomeric-repeat binding factor 2), ATM (ataxia telangiectasia mutated) and p53 have been identified involved in senescence induction, how it is triggered remains unclear. Here, we propose an integrated model associating telomere loss with senescence trigger. We characterize the dynamics of telomere shorting and the p53-centered regulatory network. We show that senescence is initiated in a switch-like manner when both the shortest telomere becomes uncapped and the TRF2-ATM-p53-Siah1 positive feedback loop is switched on. This work provides a coherent picture of senescence induction in terms of telomere shortening and p53 activation.


Change in dominance determines herbivore effects on plant biodiversity.
Herbivores alter plant biodiversity (species richness) in many of the world's ecosystems, but the magnitude and the direction of herbivore effects on biodiversity vary widely within and among ecosystems. One current theory predicts that herbivores enhance plant biodiversity at high productivity but have the opposite effect at low productivity. Yet, empirical support for the importance of site productivity as a mediator of these herbivore impacts is equivocal. Here, we synthesize data from 252 large-herbivore exclusion studies, spanning a 20-fold range in site productivity, to test an alternative hypothesis-that herbivore-induced changes in the competitive environment determine the response of plant biodiversity to herbivory irrespective of productivity. Under this hypothesis, when herbivores reduce the abundance (biomass, cover) of dominant species (for example, because the dominant plant is palatable), additional resources become available to support new species, thereby increasing biodiversity. By contrast, if herbivores promote high dominance by increasing the abundance of herbivory-resistant, unpalatable species, then resource availability for other species decreases reducing biodiversity. We show that herbivore-induced change in dominance, independent of site productivity or precipitation (a proxy for productivity), is the best predictor of herbivore effects on biodiversity in grassland and savannah sites. Given that most herbaceous ecosystems are dominated by one or a few species, altering the competitive environment via herbivores or by other means may be an effective strategy for conserving biodiversity in grasslands and savannahs globally.


A narrow open tubular column for high efficiency liquid chromatographic separation.
We report a great feature of open tubular liquid chromatography when it is run using an extremely narrow (e.g., 2 μm inner diameter) open tubular column: more than 10 million plates per meter can be achieved in less than 10 min and under an elution pressure of ca. 20 bar. The column is coated with octadecylsilane and both isocratic and gradient separations are performed. We reveal a focusing effect that may be used to interpret the efficiency enhancement. We also demonstrate the feasibility of using this technique for separating complex peptide samples. This high-resolution and fast separation technique is promising and can lead to a powerful tool for trace sample analysis.


Caffeine Enhances Memory Performance in Young Adults during Their Non-optimal Time of Day.
Many college students struggle to perform well on exams in the early morning. Although students drink caffeinated beverages to feel more awake, it is unclear whether these actually improve performance. After consuming coffee (caffeinated or decaffeinated), college-age adults completed implicit and explicit memory tasks in the early morning and late afternoon (Experiment 1). During the morning, participants ingesting caffeine demonstrated a striking improvement in explicit memory, but not implicit memory. Caffeine did not alter memory performance in the afternoon. In Experiment 2, participants engaged in cardiovascular exercise in order to examine whether increases in physiological arousal similarly improved memory. Despite clear increases in physiological arousal, exercise did not improve memory performance compared to a stretching control condition. These results suggest that caffeine has a specific benefit for memory during students' non-optimal time of day - early morning. These findings have real-world implications for students taking morning exams.


Molecular regulation of HDL metabolism and function: implications for novel therapies.
HDL metabolism represents a major target for the development of therapies intended to reduce the risk of atherosclerotic cardiovascular disease. HDL metabolism is complex and involves dissociation of HDL apolipoprotein and HDL cholesterol metabolism. Advances in our understanding of the molecular regulation of HDL metabolism, macrophage cholesterol efflux, and HDL function will lead to a variety of novel therapeutics.


A highly ordered structure in V(D)J recombination cleavage complexes is facilitated by HMG1.
Central to understanding the process of V(D)J recombination is appreciation of the protein-DNA complex which assembles on the recombination signal sequences (RSS). In addition to RAG1 and RAG2, the protein HMG1 is known to stimulate the efficiency of the cleavage reaction. Using electrophoretic mobility shift analysis we show that HMG1 stimulates the in vitro assembly of a stable complex with the RAG proteins on each RSS. We use UV crosslinking studies of this complex with azido-phenacyl derivatized probes to map the contact sites between the RAG proteins, HMG1 derivatives and the RSS. We find that the RAG proteins make contacts at the nonamer, heptamer and adjacent coding region. The HMG1 protein by itself appears to localize at the 3' side of the nonamer, but a cooperative complex with the RAG proteins is positioned at the 3' side of the heptamer and adjacent spacer in the 12RSS. In the complex with RAG proteins, HMG1 is positioned primarily in the spacer of the 23RSS. We suggest that bends introduced into these DNA substrates at specific locations by the RAG proteins and HMG1 may help distinguish the 12RSS from the 23RSS and may therefore play an important role in the coordinated reaction.


The product of the spindle formation gene sad1+ associates with the fission yeast spindle pole body and is essential for viability.
Spindle formation in fission yeast occurs by the interdigitation of two microtubule arrays extending from duplicated spindle pole bodies which span the nuclear membrane. By screening a bank of temperature-sensitive mutants by anti-tubulin immunofluorescence microscopy, we previously identified the sad1.1 mutation (Hagan, I., and M. Yanagida. 1990. Nature (Lond.). 347:563-566). Here we describe the isolation and characterization of the sad1+ gene. We show that the sad1.1 mutation affected both spindle formation and function. The sad1+ gene is a novel essential gene that encodes a protein with a predicted molecular mass of 58 kD. Deletion of the gene was lethal resulting in identical phenotypes to the sad1.1 mutation. Sequence analysis predicted a potential membrane-spanning domain and an acidic amino terminus. Sad1 protein migrated as two bands of 82 and 84 kD on SDS-PAGE, considerably slower than its predicted mobility, and was exclusively associated with the spindle pole body (SPB) throughout the mitotic and meiotic cycles. Microtubule integrity was not required for Sad1 association with the SPB. Upon the differentiation of the SPB in metaphase of meiosis II, Sad1-staining patterns similarly changed from a dot to a crescent supporting an integral role in SPB function. Moderate overexpression of Sad1 led to association with the nuclear periphery. As Sad1 was not detected in the cytoplasmic microtubule-organizing centers activated at the end of anaphase or kinetochores, we suggest that Sad1 is not a general component of microtubule-interacting structures per se, but is an essential mitotic component that associates with the SPB but is not required for microtubule nucleation. Sad1 may play a role in SPB structure, such as maintaining a functional interface with the nuclear membrane or in providing an anchor for the attachment of microtubule motor proteins.


The Recovery of Plastid Function Is Required for Optimal Response to Low Temperatures in Arabidopsis.
Cold acclimation is an essential response in higher plants to survive freezing temperatures. Here, we report that two independent mutant alleles of the H-subunit of Mg-chelatase, CHLH, gun5-1 and cch in Arabidopsis are sensitive to low temperatures. Plants were grown in photoperiodic conditions and exposed to low temperatures for short- and long-term periods. Tetrapyrrole biosynthesis was initially significantly inhibited in response to low temperature but recovered in wild type (Col-0), although the tetrapyrrole levels were lower in cold compared to control conditions. The gun5-1 and cch alleles showed an inability to recover chlorophyll biosynthesis in addition to a significant decrease in freezing tolerance. We found that the impaired plastid function in the CHLH mutant plants resulted in compromised de novo protein synthesis at low temperatures. The expression of the transcription factors CBF1-3 was super-induced in gun5-1 and cch mutant alleles but expression levels of their target genes, COR15a, COR47 and COR78 were similar or even lower compared to Col-0. In addition, the protein levels of COR15a were lower in gun5-1 and cch and a general defect in protein synthesis could be seen in the gun5-1 mutant following a 35S labelling experiment performed at low temperature. Taken together, our results demonstrate the importance of a functional chloroplast for the cold acclimation process and further suggest that impaired plastid function could result in inhibition of protein synthesis at low temperature.


Ipsilateral corticospinal projections do not predict congenital mirror movements: a case report.
Congenital mirror movements (CMMs) are involuntary, symmetric movements of one hand during the production of voluntary movements with the other. CMMs have been attributed to a range of physiological mechanisms, including excessive ipsilateral projections from each motor cortex to distal extremities. We examined this hypothesis with an individual showing pronounced CMMs. Mirror movements were characterized for a set of hand muscles during a simple contraction task. Transcranial magnetic stimulation (TMS) was then used to map the relative input to each muscle from both motor cortices. Contrary to our expectations, CMMs were most prominent for muscles with the strongest contralateral representation rather than in muscles that were activated by stimulation of either hemisphere. These findings support a bilateral control hypothesis whereby CMMs result from the recruitment of both motor cortices during intended unimanual movements. Consistent with this hypothesis, bilateral motor cortex activity was evident during intended unimanual movements in an fMRI study. To assess the level at which bilateral recruitment occurs, motor cortex excitability during imagined unimanual movements was assessed with TMS. Facilitory excitation was only observed in the contralateral motor cortex. Thus, the bilateral recruitment of the hemispheres for unilateral actions in individuals with CMMs appears to occur during movement execution rather than motor planning.


Lack of correlation between Tc-99m-sestaMIBI uptake and cadherin expression in infiltrating ductal breast carcinoma as prognostic indicators.
Despite using various kinds of prognostic indicators, it is still not possible to predict the biological behavior of breast cancer in all patients. Tc-99m-sestaMIBI (MIBI) uptake determined by breast scintigraphy and cadherin expression of tumor tissue revealed by immunohistochemistry are suggested as potential agents for this purpose. We hypothesize that there can be a correlation between MIBI whose cellular mitochondrial content is claimed to play a significant role in its tumor uptake and cadherin whose downregulation causes an increase in mitochondrial activity in human mammary carcinoma cell lines. The aim of this study was to assess the relationship between the degree of MIBI tumor uptake and cadherin expression in infiltrating ductal breast carcinoma. Correlation with response to chemotherapy and some known prognostic factors of breast cancer such as tumor size, number of metastatic axillary lymph nodes and microscopic grading was also done. Fourteen patients who underwent scintimammography and subsequent surgical excisional biopsy that revealed infiltrating ductal carcinoma were enrolled in this study. Statistical analysis did not show any correlation between MIBI uptake and cadherin expression (p > 0.05). Also, no statistically significant correlation was noted between MIBI uptake and tumor size, number of metastatic lymph nodes, microscopic grade, stage of the disease or response to chemotherapy. Similarly, there was no statistically significant correlation between cadherin expression and tumor size, number of metastatic lymph nodes, microscopic grade, stage of the disease or chemotherapy response. The results of this study imply that there is no correlation between MIBI tumor uptake and cadherin expression with neither of them good enough to be used as prognostic indicators for breast cancer.


A pulmonary embolism response team's initial 20 month experience treating 87 patients with submassive and massive pulmonary embolism.
Pulmonary Embolism Response Teams (PERTs) have emerged to provide rapid multidisciplinary assessment and treatment of PE patients. However, descriptive institutional experience and preliminary outcomes data from such teams are sparse. PERT activations were identified through a retrospective review. Only confirmed submassive or massive PEs were included in the data analysis. In addition to baseline variables, the therapeutic intervention, length of stay (LOS), in-hospital mortality, and bleeding rate/severity were recorded. A total of 124 PERT activations occurred over 20 months: 43 in the first 10 months and 81 in the next 10. A total of 87 submassive (90.8%) and massive (9.2%) PE patients were included. The median age was 65 (51-75 IQR) years. Catheter-directed thrombolysis (CDT) was administered to 25 patients, systemic thrombolysis (ST) to six, and anticoagulation alone (AC) to 54. The median ICU stay and overall LOS were 6 (3-10 IQR) and 7 (4-14 IQR) days, respectively, with no association with any variables except a brain natriuretic peptide (BNP) >100 pg/mL ( p=0.008 ICU LOS; p=0.047 overall LOS). Twelve patients (13.7%) died in the hospital, nine of whom had metastatic or brain cancer, with a median overall LOS of 13 (11-17 IQR) days. There were five major bleeds: one in the CDT group, one in the ST group, and three in the AC group. Overall, (1) PERT activations increased after the first 10 months; (2) BNP >100 pg/mL was associated with a longer LOS; (3) rates of mortality and bleeding did not correlate with treatment; and (4) the majority of in-hospital deaths occurred in patients with advanced cancer.


Genetic characterization of an H2N2 influenza virus isolated from a muskrat in Western Siberia.
Thirty-two muskrats (Ondatra zibethicus) were captured for surveillance of avian influenza virus in wild waterfowl and mammals near Lake Chany, Western Siberia, Russia. A/muskrat/Russia/63/2014 (H2N2) was isolated from an apparently healthy muskrat using chicken embryos. Based on phylogenetic analysis, the hemagglutinin and neuraminidase genes of this isolate were classified into the Eurasian avian-like influenza virus clade and closely related to low pathogenic avian influenza viruses (LPAIVs) isolated from wild water birds in Italy and Sweden, respectively. Other internal genes were also closely related to LPAIVs isolated from Eurasian wild water birds. Results suggest that interspecies transmission of LPAIVs from wild water birds to semiaquatic mammals occurs, facilitating the spread and evolution of LPAIVs in wetland areas of Western Siberia.


Antioxidant activity and mechanisms of action of natural compounds isolated from lichens: a systematic review.
Chronic diseases such as cancer, diabetes, neurodegenerative and cardiovascular diseases are characterized by an enhanced state of oxidative stress, which may result from the overproduction of reactive species and/or a decrease in antioxidant defenses. The search for new chemical entities with antioxidant profile is still thus an emerging field on ongoing interest. Due to the lack of reviews concerning the antioxidant activity of lichen-derived natural compounds, we performed a review of the antioxidant potential and mechanisms of action of natural compounds isolated from lichens. The search terms "lichens", "antioxidants" and "antioxidant response elements" were used to retrieve articles in LILACS, PubMed and Web of Science published until February 2014. From a total of 319 articles surveyed, 32 met the established inclusion and exclusion criteria. It was observed that the most common isolated compound studied was usnic acid, cited in 14 out of the 32 articles. The most often described antioxidant assays for the study of in vitro antioxidant activity were mainly DPPH, LPO and SOD. The most suggested mechanisms of action were scavenging of reactive species, enzymatic activation and inhibition of iNOS. Thus, compounds isolated from lichens are possible candidates for the management of oxidative stress, and may be useful in the treatment of chronic diseases.


Analysis of compartmentalized cAMP: a method to compare signals from differently targeted FRET reporters.
Förster resonance energy transfer (FRET)-based reporters are important tools to study the spatiotemporal compartmentalization of cyclic adenosine monophosphate (cAMP) in living cells. To increase the spatial resolution of cAMP detection, new reporters with specific intracellular targeting have been developed. Therefore it has become critical to be able to appropriately compare the signals revealed by the different sensors. Here we illustrate a protocol to calibrate the response detected by different targeted FRET reporters involving the generation of a dose-response curve to the cAMP raising agent forskolin. This method represents a general tool for the accurate analysis and interpretation of intracellular cAMP changes detected at the level of different subcellular compartments.


Assessment of Somatosensory Function in Patients With Idiopathic Dry Eye Symptoms.
Somatosensory dysfunction likely underlies dry eye (DE) symptoms in many individuals yet remains an understudied component of the disease. Its presence has important diagnostic and therapeutic implications. To assess the integrity of nociceptive system processes in persons with DE and ocular pain using quantitative sensory testing (QST) techniques applied at a site remote from the eye. A cross-sectional study conducted at Miami Veterans Affairs Hospital included 118 individuals with a wide variety of DE symptoms and signs. The study was conducted from October 31, 2013, to January 28, 2016. Individuals completed questionnaires regarding ocular symptoms (5-Item Dry Eye Questionnaire [DEQ5], Ocular Surface Disease Index [OSDI], and Neuropathic Pain Symptom Inventory modified for the eye [NPSI-E]), psychological status, and medication use and underwent an ocular surface examination. The QST metrics included measures of vibratory and thermal thresholds and cold and hot pain temporal summation (surrogate measures of central sensitization) on the forearm. Correlations among DE and ocular pain symptom severity with QST metrics measured on the forearm. The OSDI score ranges from 0 to 100, with 100 indicating the most severe DE symptoms. The DEQ5 score ranges from 0 to 22, with the highest score indicating the most severe symptoms, and the NPSI-E score ranges from 0 to 100, with the highest score indicating the most severe symptoms. Psychological state was measured with the 9-item Patient Health Questionnaire, the PTSD Checklist-Military Version for PTSD, and the Symptom Checklist-90 for anxiety. Of the 118 patients who participated in the study, 105 (88.9%) were men (mean [SD] age, 60 [10] years), and a mean of 41% had PTSD, 10% depression, and 0.93% anxiety. Using stepwise linear regression analyses, significant associations were identified between overall DE symptom severity and posttraumatic stress disorder scores and tear breakup time (DEQ5 model: R = 0.54; OSDI model: R = 0.61, P < .001). All other variables (ie, demographics, comorbidities, medications, tear film factors, and QST metrics) dropped out of these models. When specifically considering neuropathic-like qualities of DE pain, however, anxiety and hot pain temporal summation at the forearm explained 17% of the variability in ocular burning (R = 0.41; P < .001), and PTSD score, tear breakup time, and hot pain temporal summation at the forearm explained 25% of the variability in sensitivity to wind (R = 0.50; P < .001) and 30% of the variability in total NPSI-E scores (R = 0.55; P < .001). Our findings demonstrate that neuropathic-like DE pain symptom severity correlates with quantitative measures of pain sensitivity at a site remote from the eye. This result provides additional evidence that DE symptoms are not only manifestations of a local disorder but also involve somatosensory dysfunction beyond the trigeminal system.


Plasticity of directional place fields in a model of rodent CA3.
We propose a computational model of the CA3 region of the rat hippocampus that is able to reproduce the available experimental data concerning the dependence of directional selectivity of the place cell discharge on the environment and on the spatial task. The main feature of our model is a continuous, unsupervised Hebbian learning dynamics of recurrent connections, which is driven by the neuronal activities imposed upon the network by the environment-dependent external input. In our simulations, the environment and the movements of the rat are chosen to mimic those commonly observed in neurophysiological experiments. The environment is represented as local views that depend on both the position and the heading direction of the rat. We hypothesize that place cells are intrinsically directional, that is, they respond to local views. We show that the synaptic dynamics in the recurrent neural network rapidly modify the discharge correlates of the place cells: Cells tend to become omnidirectional place cells in open fields, while their directionality tends to get stronger in radial-arm mazes. We also find that the synaptic learning mechanisms account for other properties of place cell activity, such as an increase in the place cell peak firing rates as well as clustering of place fields during exploration. Our model makes several experimental predictions that can be tested using current techniques.


The relationship between pre-morbid personality and challenging behaviour in people with dementia: A systematic review.
It has been suggested that challenging behaviour in people with dementia reflects a person's pre-morbid personality traits and a number of studies have explored this hypothesis. However, inconsistencies in outcome between studies suggest a need to review the available evidence systematically. As a result, major bibliographic databases were searched for studies examining the relationship between pre-morbid personality and challenging behaviour in order to conduct a systematic review. We included all English language studies published in referenced journals that assessed pre-morbid personality via a valid comprehensive personality measure, and also explored a relationship with challenging behaviour in people with dementia. A total of 18 studies were identified that covered a wide range of challenging behaviours including 'wandering', affective states, aggression, anxiety and delusions/hallucinations. Studies were assessed for their methodological quality and statistical findings. Studies lacked representative samples, were affected by confounding variables and suffered from small sample sizes. However, 72% of the studies reported significant relationships between pre-morbid personality and behaviour. In terms of specific relationships, the strongest evidence was found for a positive relationship between pre-morbid neuroticism and mood, and aggression and overall behavioural acts, thus supporting the inclusion of personality as one factor in the formulation of behaviour (Ballard, C., O'Brien, J., James, I., & Swann, A. (2001). Dementia: Management of Behavioural and Psychological Symptoms. Oxford: Oxford University Press; Kitwood, T. (1993). Person and process in dementia: Editorial. International Journal of Geriatric Psychiatry, 1, 541-545).


Common and specific responses to availability of mineral nutrients and water.
Changes in resource (mineral nutrients and water) availability, due to their heterogeneous distribution in space and time, affect plant development. Plants need to sense these changes to optimize growth and biomass allocation by integrating root and shoot growth. Since a limited supply of water or nutrients can elicit similar physiological responses (the relative activation of root growth at the expense of shoot growth), similar underlying mechanisms may affect perception and acquisition of either nutrients or water. This review compares root and shoot responses to availability of different macronutrients and water. Attention is given to the roles of root-to-shoot signalling and shoot-to-root signalling, with regard to coordinating changes in root and shoot growth and development. Involvement of plant hormones in regulating physiological responses such as stomatal and hydraulic conductance is revealed by measuring the effects of resource availability on phytohormone concentrations in roots and shoots, and their flow between roots and shoots in xylem and phloem saps. More specific evidence can be obtained by measuring the physiological responses of genotypes with altered hormone responses or concentrations. We discuss the similarity and diversity of changes in shoot growth, allocation to root growth, and root architecture under changes in water, nitrate, and phosphorus availability, and the possible involvement of abscisic acid, indole-acetic acid, and cytokinin in their regulation. A better understanding of these mechanisms may contribute to better crop management for efficient use of these resources and to selecting crops for improved performance under suboptimal soil conditions.


Epidemiology of Acute Kidney Injury in Critically Ill Children and Young Adults.
The epidemiologic characteristics of children and young adults with acute kidney injury have been described in single-center and retrospective studies. We conducted a multinational, prospective study involving patients admitted to pediatric intensive care units to define the incremental risk of death and complications associated with severe acute kidney injury. We used the Kidney Disease: Improving Global Outcomes criteria to define acute kidney injury. Severe acute kidney injury was defined as stage 2 or 3 acute kidney injury (plasma creatinine level ≥2 times the baseline level or urine output <0.5 ml per kilogram of body weight per hour for ≥12 hours) and was assessed for the first 7 days of intensive care. All patients 3 months to 25 years of age who were admitted to 1 of 32 participating units were screened during 3 consecutive months. The primary outcome was 28-day mortality. A total of 4683 patients were evaluated; acute kidney injury developed in 1261 patients (26.9%; 95% confidence interval [CI], 25.6 to 28.2), and severe acute kidney injury developed in 543 patients (11.6%; 95% CI, 10.7 to 12.5). Severe acute kidney injury conferred an increased risk of death by day 28 after adjustment for 16 covariates (adjusted odds ratio, 1.77; 95% CI, 1.17 to 2.68); death occurred in 60 of the 543 patients (11.0%) with severe acute kidney injury versus 105 of the 4140 patients (2.5%) without severe acute kidney injury (P<0.001). Severe acute kidney injury was associated with increased use of mechanical ventilation and renal-replacement therapy. A stepwise increase in 28-day mortality was associated with worsening severity of acute kidney injury (P<0.001 by log-rank test). Assessment of acute kidney injury according to the plasma creatinine level alone failed to identify acute kidney injury in 67.2% of the patients with low urine output. Acute kidney injury is common and is associated with poor outcomes, including increased mortality, among critically ill children and young adults. (Funded by the Pediatric Nephrology Center of Excellence at Cincinnati Children's Hospital Medical Center and others; AWARE ClinicalTrials.gov number, NCT01987921 .).


MicroRNA Biogenesis Pathway Genes Are Deregulated in Colorectal Cancer.
MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression. Each step of their production and maturation has to be strictly regulated, as any disruption of control mechanisms may lead to cancer. Thus, we have measured the expression of 19 genes involved in miRNAs biogenesis pathway in tumor tissues of 239 colorectal cancer (CRC) patients, 17 CRC patients with liver metastases and 239 adjacent tissues using real-time PCR. Subsequently, the expression of analyzed genes was correlated with the clinical-pathological features as well as with the survival of patients. In total, significant over-expression of all analyzed genes was observed in tumor tissues as well as in liver metastases except for LIN28A/B. Furthermore, it was shown that the deregulated levels of some of the analyzed genes significantly correlate with tumor stage, grade, location, size and lymph node positivity. Finally, high levels of DROSHA and TARBP2 were associated with shorter disease-free survival, while the over-expression of XPO5, TNRC6A and DDX17 was detected in tissues of patients with shorter overall survival and poor prognosis. Our data indicate that changed levels of miRNA biogenesis genes may contribute to origin as well as progression of CRC; thus, these molecules could serve as potential therapeutic targets.


DeepDrug3D: Classification of ligand-binding pockets in proteins with a convolutional neural network.
Comprehensive characterization of ligand-binding sites is invaluable to infer molecular functions of hypothetical proteins, trace evolutionary relationships between proteins, engineer enzymes to achieve a desired substrate specificity, and develop drugs with improved selectivity profiles. These research efforts pose significant challenges owing to the fact that similar pockets are commonly observed across different folds, leading to the high degree of promiscuity of ligand-protein interactions at the system-level. On that account, novel algorithms to accurately classify binding sites are needed. Deep learning is attracting a significant attention due to its successful applications in a wide range of disciplines. In this communication, we present DeepDrug3D, a new approach to characterize and classify binding pockets in proteins with deep learning. It employs a state-of-the-art convolutional neural network in which biomolecular structures are represented as voxels assigned interaction energy-based attributes. The current implementation of DeepDrug3D, trained to detect and classify nucleotide- and heme-binding sites, not only achieves a high accuracy of 95%, but also has the ability to generalize to unseen data as demonstrated for steroid-binding proteins and peptidase enzymes. Interestingly, the analysis of strongly discriminative regions of binding pockets reveals that this high classification accuracy arises from learning the patterns of specific molecular interactions, such as hydrogen bonds, aromatic and hydrophobic contacts. DeepDrug3D is available as an open-source program at https://github.com/pulimeng/DeepDrug3D with the accompanying TOUGH-C1 benchmarking dataset accessible from https://osf.io/enz69/.


Health plan members' views on forgiving medical errors.
How patients respond to medical errors may influence how physicians approach disclosure of medical errors, but information on patients' responses is limited. Research is needed on how the circumstances that surround a medical error affect how patients respond. To investigate whether patients' tendency to forgive a physician following a medical error varied under different circumstances. Cross-sectional survey. We mailed a questionnaire to 1500 randomly selected health plan members; the response rate was 66%. Questionnaire items assessed the likelihood of forgiveness following a medical error under 12 circumstances drawn from a review of the literature. Respondents were most likely to forgive a physician if the patient failed to provide complete information (93% would or might forgive) and least likely to forgive if the error was due to efforts to keep costs down (11% would or might forgive). Most respondents would not forgive a physician when the physician was tired or distracted (68%), was incomplete in data collection (76%), lacked knowledge (78%), or failed to follow up (85%). Men were more likely to forgive than women; the most educated respondents were most likely to forgive. Our findings suggest that patients are not likely to forgive a physician in circumstances in which they suspect incompetence, inattention, or a lack of caring on the part of the physician involved. A more comprehensive understanding of forgiveness and the effect of forgiveness on the physician-patient relationship following a medical error is needed.


Correlation of Fractional Limb Volume Measurements with Neonatal Morphometric Indices.
Fractional thigh volume (TVol) and fractional arm volume (AVol) measurements by three-dimensional (3D) ultrasound can reveal valuable information on fetal soft tissue development. However, it is not clear whether TVol or AVol provides better estimates of fetal body proportion and adiposity, independent of routine two-dimensional (2D) ultrasound biometry. The primary objective of the current study was to determine the correlations between fractional limb volumes (FLVs) and neonatal anthropometric parameters. In this cross-sectional study, fetal FLVs were obtained within 24 h before term delivery from 40 medically and obstetrically uncomplicated pregnancies scheduled for elective cesarean section. TVol and AVol were determined using offline software. Postnatal morphometric data including birth weight; crown-heel, arm, and leg lengths; head, abdominal, mid-thigh, and mid-arm circumferences; and anterior thigh, biceps, and subscapular skinfold thicknesses were obtained. Pearson and partial correlation analyses were used to determine the relationships across antenatal volume calculations and neonatal indices. Correlation coefficients (r) were calculated. Mean maternal age, BMI, and parity were 29.1 ± 5.4 years, 29.7 ± 3.5 kg/m2, and 1.0 ± 1.3, respectively. AVol showed moderate correlations with most of the neonatal parameters, including mid-thigh circumference (r = 0.683), mid-arm circumference (r = 0.627), birth weight (r = 0.583), head circumference (HC, r = 0.560), and abdominal circumference (r = 0.542). However, TVol was weakly related to only some of the indices. After controlling for gestational age, maternal age, BMI, parity, and 2D ultrasound biometry, TVol was no longer associated with any of the parameters, while AVol was independently correlated with mid-thigh (r = 0.724) and mid-arm circumference (r = 0.560), birth weight (r = 0.502), ponderal index (r = 0.402), HC (r = 0.382), biceps (r = 0.384), and subscapular skinfold thickness (r = 0.350). The current design includes limited number of pregnancies with only scheduled cesarean deliveries. Neonatal percent body fat was not calculated, and air-displacement plethysmography was not used to assess neonatal body composition. The study population was Caucasian with a relatively high maternal BMI, which may limit extrapolation of the results to other settings. AVoL measurements by 3D ultrasound before delivery are significantly correlated with most of the neonatal morphometric indices, independent of maternal characteristics and 2D biometric parameters. AVol may have advantages over TVol for assessing limb soft tissue development in term fetuses. Future research can focus on feasibility and predictive ability of AVol measurements in prospective studies that include serial biometry over time.


Craniosynostosis of coronal suture in twist1 mice occurs through endochondral ossification recapitulating the physiological closure of posterior frontal suture.
Craniosynostosis, the premature closure of cranial suture, is a pathologic condition that affects 1/2000 live births. Saethre-Chotzen syndrome is a genetic condition characterized by craniosynostosis. The Saethre-Chotzen syndrome, which is defined by loss-of-function mutations in the TWIST gene, is the second most prevalent craniosynostosis. Although much of the genetics and phenotypes in craniosynostosis syndromes is understood, less is known about the underlying ossification mechanism during suture closure. We have previously demonstrated that physiological closure of the posterior frontal suture occurs through endochondral ossification. Moreover, we revealed that antagonizing canonical Wnt-signaling in the sagittal suture leads to endochondral ossification of the suture mesenchyme and sagittal synostosis, presumably by inhibiting Twist1. Classic Saethre-Chotzen syndrome is characterized by coronal synostosis, and the haploinsufficient Twist1(+/-) mice represents a suitable model for studying this syndrome. Thus, we seeked to understand the underlying ossification process in coronal craniosynostosis in Twist1(+/-) mice. Our data indicate that coronal suture closure in Twist1(+/-) mice occurs between postnatal day 9 and 13 by endochondral ossification, as shown by histology, gene expression analysis, and immunohistochemistry. In conclusion, this study reveals that coronal craniosynostosis in Twist1(+/-) mice occurs through endochondral ossification. Moreover, it suggests that haploinsufficiency of Twist1 gene, a target of canonical Wnt-signaling, and inhibitor of chondrogenesis, mimics conditions of inactive canonical Wnt-signaling leading to craniosynostosis.


Comparative permeability of fresh and frozen/thawed porcine buccal mucosa towards various chemical markers.
Sometimes it is not feasible or convenient to use tissue samples immediately and they are then stored at -85 degrees C after being snap-frozen in liquid N2. Previous studies have shown that, although snap-freezing in liquid N2 does not alter the permeability properties of human buccal and vaginal mucosae towards various markers significantly, some tissue damage does occur. Slight increases in the permeability properties of the frozen/thawed human buccal mucosa towards markers were found, although these were not statistically significant. The damage does, however, not seem to affect the lipid permeability barrier to a meaningful extent. Because porcine buccal mucosa is a popular choice as a substitute for human buccal mucosa in many permeability studies, it was decided to evaluate the effect that snap-freezing, storage and thawing would have on the permeability of this mucosa when compared to human mucosae. Fresh and frozen/thawed buccal mucosa specimens were collected from freshly slaughtered pigs and permeability experiments at 20 degrees C and 1.5 ml/h, over a 24 hour time period, performed using a flow-through diffusion apparatus. The permeants tested were water, arecoline, vasopressin and 17beta-estradiol. Statistically significant differences were obtained between the flux values at steady state for water, arecoline and vasopressin through fresh and frozen/thawed porcine buccal mucosa. The estimated mean steady state flux values for 17beta-estradiol showed no statistically significant differences between the fresh appear to indicate that the lipid barrier of porcine buccal mucosa is not as resistant to snap-freezing, storage and thawing procedures as that of human mucosal tissue.


Control of Walking Speed in Children With Cerebral Palsy.
Children's ability to control the speed of gait is important for a wide range of activities. It is thought that the ability to increase the speed of gait for children with cerebral palsy (CP) is common. This study considered 3 hypotheses: (1) most ambulatory children with CP can increase gait speed, (2) the characteristics of free (self-selected) and fast walking are related to motor impairment level, and (3) the strategies used to increase gait speed are distinct among these levels. A retrospective review of time-distance parameters (TDPs) for 212 subjects with CP and 34 typically developing subjects walking at free and fast speeds was performed. Only children who could increase their gait speed above the minimal clinically important difference were defined as having a fast walk. Analysis of variance was used to compare TDPs of children with CP, among Gross Motor Function Classification System (GMFCS) levels, and children in typically developing group. Eight-five percent of the CP group (GMFCS I, II, III; 96%, 99%, and 34%, respectively) could increase gait speed on demand. At free speed, children at GMFCS I and II were significantly faster than children at GMFCS level III. At free speed, children at GMFCS I and II had significantly greater stride length than those at GMFCS levels III. At free speed, children at GMFCS level III had significantly lower cadence than those at GMFCS I and II. There were no significant differences in cadence among GMFCS levels at fast speeds. There were no significant differences among GMFCS levels for percent change in any TDP between free and fast walking. Almost all children with CP at GMFCS levels I and II can control the speed of gait, however, only one-third at GMFCS III level have this ability. This study suggests that children at GMFCS III level can be divided into 2 groups based on their ability to control gait speed; however, the prognostic significance of such categorization remains to be determined. Diagnostic level II.


Clinical-surgical treatment of temporomandibular joint disorder in a psoriatic arthritis patient.
Condylotomy is a surgical procedure that has been used as an option to treat temporomandibular disorder (TMD) patients. This technique has the advantage of avoiding intra-capsular alterations that might be found involving other surgical procedures. Its use, even when unilateral, has positive effect on treatment of both joints. In order to better evaluate the benefits of a clinical-surgical treatment for TMD, the present report describes the case of a psoriatic arthritis patient. The case was clinically characterized by dental malloclusion, and imaging exams showed joint degeneration of the right mandibular condyle. The patient was treated by condylotomy technique after a prosthetic oral rehabilitation. No clinical-radiological signs or symptoms of progression of articular disease were observed within a period of 16 months after surgery. Furthermore, there was functional stability of the temporomandibular joint, total absence of local pain and improvement of mouth opening. The present study suggests that condylotomy can be considered as a valid option for the management of TMD, since it has low surgical morbidity and favorable clinical outcomes. In this case, the patient had a medical diagnosis of systemic disease presenting general pain and pain at the temporomandibular joint (TMJ), in addition of causal agent of TMD (dental malloclusion). The difficulty of finding a single etiology (malocclusion vs. systemic disease) did not exclude the indication of a clinical-surgical treatment to re-establish the balance of TMJ.


Time-motion analysis of elite male kickboxing competition.
The objective of the study was to analyze the time structure of high-level kickboxing matches. A total of 45 combats from 2 male World Championships were monitored using a time-motion analysis system. The combat time structure (i.e., high-intensity activity [HIA], low-intensity activity [LIA], and referee breaks or pauses) during competition and weight divisions was determined and compared. Results indicated that the time structures were HIA: 2.2 ± 1.2 seconds; LIA: 2.3 ± 0.8 seconds; pauses: 5.4 ± 4.3 seconds; and 3.4 ± 1.2 seconds between 2 subsequent HIA. The fighting to nonfighting ratio was found to be 1:1. Moreover, the number of HIA and LIA and the time of LIA decreased in latter rounds (e.g., the average number of HIA was 27.1 ± 7.1, 25.1 ± 6.6, and 24.9 ± 6.1, respectively, for rounds 1, 2, and 3), meanwhile the time and number of pauses increased (e.g., the average pause times were 12.8 ± 11.4, 22.3 ± 22.6, and 24.6 ± 23.3 seconds, respectively, for rounds 1, 2, and 3). The activity times did not differ among weight categories. The present results confirm the intermittent nature of kickboxing competition and provide coaches with more information on how to structure training sessions to mimic the physical demands in competition.


Clinical benefit of remdesivir in rhesus macaques infected with SARS-CoV-2.
Effective therapeutics to treat COVID-19 are urgently needed. Remdesivir is a nucleotide prodrug with in vitro and in vivo efficacy against coronaviruses. Here, we tested the efficacy of remdesivir treatment in a rhesus macaque model of SARS-CoV-2 infection. To evaluate the effect of remdesivir treatment on SARS-CoV-2 disease outcome, we used the recently established rhesus macaque model of SARS-CoV-2 infection that results in transient lower respiratory tract disease. Two groups of six rhesus macaques were infected with SARS-CoV-2 and treated with intravenous remdesivir or an equal volume of vehicle solution once daily. Clinical, virological and histological parameters were assessed regularly during the study and at necropsy to determine treatment efficacy. In contrast to vehicle-treated animals, animals treated with remdesivir did not show signs of respiratory disease and had reduced pulmonary infiltrates on radiographs. Virus titers in bronchoalveolar lavages were significantly reduced as early as 12hrs after the first treatment was administered. At necropsy on day 7 after inoculation, lung viral loads of remdesivir-treated animals were significantly lower and there was a clear reduction in damage to the lung tissue. Therapeutic remdesivir treatment initiated early during infection has a clear clinical benefit in SARS-CoV-2-infected rhesus macaques. These data support early remdesivir treatment initiation in COVID-19 patients to prevent progression to severe pneumonia.


Congenital Esophageal Atresia and Microtia in a Newborn Secondary to Mycophenolate Mofetil Exposure During Pregnancy: A Case Report and Review of the Literature.
BACKGROUND Mycophenolate mofetil (MMF) is one of the most commonly prescribed drugs to prevent organ transplant rejection in combination with calcineurin inhibitors and steroids. It has a different toxicity profile than tacrolimus and cyclosporine. Gastrointestinal tract disturbances are the most common adverse effects. The use of MMF in pregnant women, however, holds great risk of miscarriage and fetal development defects such as external ear malformation, ocular anomalies, cleft lip and palate, and abnormality of distal limbs, heart, esophagus, and kidneys. Based on post-marketing studies, its pregnancy category was reclassified as category D by the US FDA in 2007. CASE REPORT A 20-year-old woman received a deceased-donor liver transplant for end-stage liver disease secondary to autoimmune hepatitis. She had 3 miscarriages while on MMF. In her fourth pregnancy she was exposed to MMF in the first trimester, which was stopped by week 20 of the pregnancy. Obstetric ultrasound suggested a cephalic presentation fetus with abdominal circumference. Her pregnancy resulted in an infant with tracheoesophageal fistula, esophageal atresia, and a bilateral ear canal atresia (microtia) with normal sensorineural conduction. There were no other congenital abnormalities. Thoracoscopic ligation of fistula and thoracotomy with esophageal repair were performed and a bone-anchored hearing aid for conductive hearing loss was implanted. Here, we report a case of congenital esophageal atresia and microtia secondary to mycophenolate mofetil. CONCLUSIONS MMF should be avoided during pregnancy. Transplanted female patients of reproductive age should receive appropriate counseling.


Irritable bowel syndrome, its cognition, anxiety sensitivity, and anticipatory anxiety in panic disorder patients.
The present study examined the effect of irritable bowel syndrome (IBS), cognitive appraisal of IBS, and anxiety sensitivity on anticipatory anxiety (AA) and agoraphobia (AG) in patients with panic disorder (PD). We examined 244 PD patients who completed a set of questionnaires that included the Rome II Modular Questionnaire to assess the presence of IBS, the Anxiety Sensitivity Index (ASI), the Cognitive Appraisal Rating Scale (CARS; assessing the cognitive appraisal of abdominal symptoms in four dimensions: commitment, appraisal of effect, appraisal of threat, and controllability), and items about the severity of AA and AG. The Mini International Neuropsychiatric Interview was used to diagnose AG and PD. After excluding individuals with possible organic gastrointestinal diseases by using 'red flag items,' valid data were obtained from 174 participants, including 110 PD patients without IBS (PD/IBS[-]) and 64 with IBS (PD/IBS[+]). The PD/IBS[+] group had higher AA and higher comorbidity with AG than the PD/IBS[-] group. In the PD/IBS[+] group, the controllability score of CARS was significantly correlated with AA and ASI. Multiple regression analysis showed a significant effect of ASI but not of controllability on AA in PD/IBS[+] subjects. This study suggested that the presence of IBS may be related to agoraphobia and anticipatory anxiety in PD patients. Cognitive appraisal could be partly related to anticipatory anxiety in PD patients with IBS with anxiety sensitivity mediating this correlation.


No evidence for leaf-trait dissimilarity effects on litter decomposition, fungal decomposers, and nutrient dynamics.
Biodiversity and ecosystem-functioning theory suggest that litter mixtures composed of dissimilar leaf species can enhance decomposition due to species trait complementarity. Here we created a continuous gradient of litter chemistry trait variability within species mixtures to assess effects of litter dissimilarity on three related processes in a natural stream: litter decomposition, fungal biomass accrual in the litter, and nitrogen and phosphorus immobilization. Litter from a pool of eight leaf species was analyzed for chemistry traits affecting decomposition (lignin, nitrogen, and phosphorus) and assembled in all of the 28 possible two-species combinations. Litter dissimilarity was characterized in terms of a range of trait-diversity measures, using Euclidean and Gower distances and dendrogram-based indices. We found large differences in decomposition rates among leaf species, but no significant relationships between decomposition rate of individual leaf species and litter trait dissimilarity, irrespective of whether decomposition was mediated by microbes alone or by both microbes and litter-consuming invertebrates. Likewise, no effects of trait dissimilarity emerged on either fungal biomass accrual or changes during decomposition of nitrogen or phosphorus concentrations in individual leaf species. In line with recent meta-analyses, these results provide support for the contention that litter diversity effects on decomposition, at least in streams, are less pronounced than effects on terrestrial primary productivity.


Combination therapy with sulfasalazine and methotrexate is more effective than either drug alone in patients with rheumatoid arthritis with a suboptimal response to sulfasalazine: results from the double-blind placebo-controlled MASCOT study.
Optimal use of disease-modifying antirheumatic drugs (DMARDs) in rheumatoid arthritis is vital if progression of disease is to be reduced. Methotrexate (MTX) and sulfasalazine (SASP) are widely used inexpensive DMARDs, recently often combined despite no firm evidence of benefit from previous studies. To establish whether a combination of SASP and MTX is superior to either drug alone in patients with rheumatoid arthritis with a suboptimal response to 6 months of SASP. A randomised controlled study of step-up DMARD treatment in early rheumatoid arthritis. In phase I, 687 patients received SASP for 6 months. Those with a disease activity score (DAS) > or =2.4 were offered additional treatment in phase II (SASP alone, MTX alone or a combination of the two). The primary outcome measure was change in DAS. At 6 months, 191 (28%) patients had a DAS <2.4, 123 (18%) were eligible but did not wish to enter phase II, 130 (19%) stopped SASP because of reversible adverse events and 165 (24%) entered phase II. DAS at 18 months was significantly lower in those who received combination treatment compared with those who received either SASP or MTX: monotherapy arms did not differ. Improvement in European League Against Rheumatism and American College of Rheumatology 20, 50 and 70 scores favoured combination therapy. In this "true-to-life" study, an inexpensive combination of DMARDs proved more effective than monotherapy in patients with rheumatoid arthritis with a suboptimal response to SASP. There was no increase in toxicity. These results provide an evidence base for the use of this combination as a component of tight control strategies.


Transversus Abdominis Plane Block Versus Surgical Site Infiltration for Pain Management After Open Total Abdominal Hysterectomy.
Surgical site infiltration and transversus abdominis plane (TAP) blocks are commonly used to improve pain relief after lower abdominal surgery. This randomized, observer-blinded study was designed to compare the analgesic efficacy of TAP blocks with surgical site infiltration in patients undergoing open total abdominal hysterectomy via a Pfannenstiel incision. Patients were randomized to receive either bilateral ultrasound-guided TAP blocks using bupivacaine 0.5% 20 mL on each side (n = 30) or surgical site infiltration with liposomal bupivacaine 266 mg diluted to 60 mL injected in the preperitoneal, subfascial, and subcutaneous planes (n = 30). The remaining aspects of the perioperative care were standardized. An investigator blinded to the group allocation documented pain scores at rest and with coughing, opioid requirements, nausea, vomiting, and rescue antiemetics in the postanesthesia care unit and at 2, 6, 12, 24, and 48 hours postoperatively. The primary outcome measure was pain scores on coughing at 6 hours postoperatively. One patient in each group was excluded from the analysis because of reoperation within 24 hours in the TAP block group and change of incision type in the infiltration group. The pain scores at rest and with coughing were significantly lower in the surgical site infiltration group at all postoperative time points (P < 0.0001) except at rest in the postanesthesia care unit. The opioid requirements between 24 and 48 hours were significantly lower in the infiltration group (P = 0.009). The nausea scores, occurrence of vomiting, and need for rescue antiemetics were similar. Surgical site infiltration provided superior pain relief at rest and on coughing, as well as reduced opioid consumption for up to 48 hours. Future studies need to compare TAP blocks with liposomal bupivacaine with surgical site infiltration with liposomal bupivacaine.


A 2-year Double-Blind RCT Follow-up Study with Fermented Papaya Preparation (FPP) Modulating Key Markers in Middle-Age Subjects with Clustered Neurodegenerative Disease-Risk Factors.
In recent years a number of studies have reported the significant relationship between metabolic syndrome and neurodegenerative disease. There is accumulating evidence that the interplay of combined genetic and environmental risk factors (from diet to life style to pollutants) to intrinsic age-related oxi-inflammatory changes may be advocated for to explain the pandemic of neurodegenerative diseases. In recent years a specific Fermented Papaya Preparation (FPP) has been shown to significantly affect a number of redox signalling abnormalities in a variety of chronic diseases and as well in aging mechanisms either on experimental and on clinical ground. The aim of the present study was to evaluate FPP use in impending metabolic disease patients with potentially neurodegenerative disease clustered risk factors. The study population consisted of 90 patients aged 45-65 years old, with impending metabolic syndrome and previously selected as to be ApoE4 genotype negative. By applying a RCT, double-blind method, one group received FPP 4.5 g twice a day (the most common dosage utilized in prior clinical studies) while the other received an oral antioxidant cocktail (trans-resveratrol, selenium, vitamin E, vitamin C). Then, after 21 month treatment period, a selected heavy metal chelator was added at the dosage of 3 g/nocte for the final 3 months study treatment. The parameters tested were: routine tests oxidized LDL-cholesterol, anti-oxidised LDL, Cyclophilin-A (CyPA), plasminogen activator inhibitor-1 and CyPA gene expression. From this study it would appear that FPP, unlike the control antioxidant, significantly decreased oxidized-LDL and near normalizing the anti-Ox-LDL/Ox-LDL ratio (p<0.001) although unaffecting the lipid profile per sè. Moreover, only FPP decreased cyclophilin-A plasma level and plasminogen activator-inhibitor (p<0.01) together with downregulating cyclophilin-A gene expression (p<0.01). Insulin resistance was only mildly improved. Heavy metals gut clearance proved to be effectively enhanced by the chelator (p<0.01) and this was not affected by any of the nutraceuticals, nor it added any further benefit to the biological action of FPP.


Molecular characterization and expression of the stratification-related cytokeratins 4 and 15.
A number of human cytokeratins are expressed during the development of stratified epithelia from one-layered polar epithelia and continue to be expressed in several adult epithelial tissues. For studies of the regulation of the synthesis of stratification-related cytokeratins in internal tissues, we have prepared cDNA and genomic clones encoding cytokeratin 4, as a representative of the basic (type II) cytokeratin subfamily and cytokeratin 15, as representative of the acidic (type I) subfamily, and determined their nucleotide sequences. The specific expression of mRNAs encoding these two polypeptides in certain stratified tissues and cultured cell lines is demonstrated by Northern blot hybridization. Hybridization in situ with antisense riboprobes and/or synthetic oligonucleotides shows the presence of cytokeratin 15 mRNA in all layers of esophagus, whereas cytokeratin 4 mRNA tends to be suprabasally enriched, although to degrees varying in different regions. We conclude that the expression of the genes encoding these stratification-related cytokeratins starts already in the basal cell layer and does not depend on vertical differentiation and detachment from the basal lamina. Our results also show that simple epithelial and stratification-related cytokeratins can be coexpressed in basal cell layers of certain stratified epithelia such as esophagus. Implications of these findings for epithelial differentiation and the formation of squamous cell carcinomas are discussed.


Zinc supplementation increases zinc status and thymopoiesis in aged mice.
The age-related decline in lymphocyte development and function coincides with impaired zinc status in the elderly. Thymic involution and reduced immune responsiveness are classic hallmarks of both aging and zinc deficiency, resulting in decreased host defense and an increased susceptibility to infections. Thus, compromised zinc status associated with aging may be an important contributing factor in reduced thymopoiesis and impaired immune functions. Our goal in this study was to understand how dietary zinc supplementation affects thymopoiesis in aged mice. We hypothesized that impaired zinc status associated with aging would mediate the decline in thymic function and output and that restoring plasma zinc concentrations via zinc supplementation would improve thymopoiesis and thymic functions. In this study, groups of young (8 wk) and aged (22 mo) mice were fed a zinc-adequate (30 mg/kg zinc) or zinc-supplemented diet (300 mg/kg) for 25 d. Aged mice had impaired zinc status, with zinc supplementation restoring plasma zinc to a concentration not different from those of young male C57Bl/6 mice. Zinc supplementation in aged mice improved thymopoiesis, as assessed by increased total thymocyte numbers. In addition, improved thymic output was mediated in part by reducing the age-related accumulation of immature CD4(-)CD8(-)CD44(+)CD25(-) thymocytes, as well as by decreasing the expression of stem cell factor, a thymosuppressive cytokine. Taken together, our results showed that in mice, zinc supplementation can reverse some age-related thymic defects and may be of considerable benefit in improving immune function and overall health in elderly populations.


Safety of TNF-α inhibitors during IBD pregnancy: a systematic review.
Tumor necrosis factor (TNF)-α inhibitors are increasingly being used in inflammatory bowel disease (IBD). Because this chronic intestinal disorder often affects women of fertile age, it is essential to assess the effect of biologics on pregnancy outcome. We performed a systematic review of the English-language literature to investigate if treatment with TNF-α blockers during pregnancy in women with IBD increases the risk of spontaneous abortions, preterm delivery, stillbirth, low birth weight, congenital malformations, or risk of infections in the offspring. Of 552 articles and abstracts reviewed, 58 articles or abstracts with unique content were identified and included in this systematic review. However, most presentations were case reports or case series supplied by a limited number of observational studies. No randomized controlled studies were available. TNF-α inhibitors do not seem to affect either outcome of pregnancy in mothers with IBD, or the outcome in the offspring (congenital malformations and immunosuppression). Further, recent data have not identified any increased risk of infections in the first year of life in the offspring of mothers who received biologics, even in combination with immunomodulators (thiopurines). From the present systematic review, no association was found between administration of TNF inhibitors for IBD during pregnancy and adverse pregnancy outcome or congenital abnormalities. Further, no increased relative risk of infections has been reported in the first year of life in offspring of mothers who received biologics. Biologics should be discontinued during pregnancy solely if the IBD is in remission using the same stopping criteria as for patients with IBD in general, as uncontrolled activity of IBD may expose the mother and child to a risk greater than those only potentially coming from the use of TNF-α inhibitors. In such cases, inoculation of the offspring with live vaccines is contraindicated until the biologic agent is no longer detectable in the child's circulation.


Skeletal muscle myoblasts possess a stretch-responsive local angiotensin signalling system.
A paucity of information exists regarding the presence of local renin-angiotensin systems (RASs) in skeletal muscle and associated muscle stem cells. Skeletal muscle and muscle stem cells were isolated from C57BL/6 mice and examined for the presence of a local RAS using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC), Western blotting and liquid chromatography-mass spectrometry (LC-MS). Furthermore, the effect of mechanical stimulation on RAS member gene expression was analysed. Whole skeletal muscle, primary myoblasts and C2C12 derived myoblasts and myotubes differentially expressed members of the RAS including angiotensinogen, angiotensin-converting enzyme (ACE), angiotensin II (Ang II) type 1 (AT(1)) and type 2 (AT(2)). Renin transcripts were never detected, however, mRNA for the 'renin-like' enzyme cathepsin D was observed and Ang I and Ang II were identified in cell culture supernatants from proliferating myoblasts. AT(1) appeared to co-localise with polymerised actin filaments in proliferating myoblasts and was primarily found in the nucleus of terminally differentiated myotubes. Furthermore, mechanical stretch of proliferating and differentiating C2C12 cells differentially induced mRNA expression of angiotensinogen, AT(1) and AT(2). Proliferating and differentiated muscle stem cells possess a local stress-responsive RAS in vitro. The precise function of a local RAS in myoblasts remains unknown. However, evidence presented here suggests that Ang II may be a regulator of skeletal muscle myoblasts.


Clinical implications of aldo-keto reductase family 1 member C3 and its relationship with lipocalin 2 in cancer of the uterine cervix.
Over-expression of the aldo-keto reductase family 1 member C3 (AKR1C3) has been demonstrated in many human cancers. Lipocalin 2 (LCN2) is reported to inhibit cervical cancer metastasis but little is known regarding its relationship with AKR1C3 in the development and progression of uterine cervical cancer. This study aimed to investigate the involvement of AKR1C3 and its relationship with LCN2 in cervical cancer. The roles of AKR1C3 and LCN2 were investigated using the lentivirus shRNA system in SiHa and Caski cervical cancer cells. LCN2 and matrix metalloproteinase-2 (MMP-2) promoters were constructed to demonstrate transcriptional regulation by shAKR1C3 and shLCN2, respectively. The influences of metastatic phenotypes were analyzed by wound healing, Boyden chamber, and immunofluorescence assays. The activity of MMP-2 was determined by zymography assay. The impacts of AKR1C3 and LCN2 on patient prognosis were evaluated using tissue microarrays by Cox regression and Kaplan-Meier models. Silencing of the AKR1C3 gene increased the expression of LCN2 and decreased the migratory and invasive abilities and changed the cytoskeleton of cervical cancer cells. When AKR1C3 was over-expressed, it decreased LCN2 promoter activity and LCN2 expression and increased cell migration. The mRNA level and enzyme activity of MMP-2 increased in silenced LCN2 cells. Positive AKR1C3 and negative LCN2 were correlated with higher recurrence and poorer survival of cervical cancer patients. Silencing of AKR1C3 increases LCN2 expression and inhibits metastasis in cervical cancer. Both AKR1C3 and LCN2 serve as molecular targets for cancer therapy to improve the clinical outcome of cervical cancer patients.


Taste responses in mice lacking taste receptor subunit T1R1.
The T1R1 receptor subunit acts as an umami taste receptor in combination with its partner, T1R3. In addition, metabotropic glutamate receptors (brain and taste variants of mGluR1 and mGluR4) are thought to function as umami taste receptors. To elucidate the function of T1R1 and the contribution of mGluRs to umami taste detection in vivo, we used newly developed knock-out (T1R1(-/-)) mice, which lack the entire coding region of the Tas1r1 gene and express mCherry in T1R1-expressing cells. Gustatory nerve recordings demonstrated that T1R1(-/-) mice exhibited a serious deficit in inosine monophosphate-elicited synergy but substantial residual responses to glutamate alone in both chorda tympani and glossopharyngeal nerves. Interestingly, chorda tympani nerve responses to sweeteners were smaller in T1R1(-/-) mice. Taste cell recordings demonstrated that many mCherry-expressing taste cells in T1R1(+/-) mice responded to sweet and umami compounds, whereas those in T1R1(-/-) mice responded to sweet stimuli. The proportion of sweet-responsive cells was smaller in T1R1(-/-) than in T1R1(+/-) mice. Single-cell RT-PCR demonstrated that some single mCherry-expressing cells expressed all three T1R subunits. Chorda tympani and glossopharyngeal nerve responses to glutamate were significantly inhibited by addition of mGluR antagonists in both T1R1(-/-) and T1R1(+/-) mice. Conditioned taste aversion tests demonstrated that both T1R1(-/-) and T1R1(+/-) mice were equally capable of discriminating glutamate from other basic taste stimuli. Avoidance conditioned to glutamate was significantly reduced by addition of mGluR antagonists. These results suggest that T1R1-expressing cells mainly contribute to umami taste synergism and partly to sweet sensitivity and that mGluRs are involved in the detection of umami compounds.


In vitro evidence for an inhibitor of lipogenesis in serum from overfed obese rats.
Involvement of a humoral agent in regulation of energy balance has been demonstrated by parabiosis experiments. Overfed obese rats produce a blood-borne factor that inhibits adipose fatty acid synthesis in their partners, resulting in loss of body fat without significant inhibition of food intake. An in vitro bioassay was developed to test small serum samples for antilipogenic activity. Epididymal adipocytes from ad libitum-fed rats were preincubated for 12 h with 2% serum. Basal adipocyte fatty acid synthesis, measured in a subsequent serum-free incubation, was inhibited by obese serum. Insulin response was not changed. Characterization studies indicated that the factor was probably a protein, larger than 30 kDa, but not a protease or a low-density lipoprotein and was not associated with serum albumin. Physiological experiments demonstrated that the agent was produced when body weight was raised substantially above "set-point." Inhibitory activity was neither species specific nor pituitary dependent. Structure, origin, and physiological significance of the factor are unknown, but it may be involved in the control of body fat content.


A Qualitative Study on the Impact of First Steps-A Peer-led Educational Intervention for People Newly Diagnosed with Parkinson's Disease.
Aim: The dual aim of this research was to consider the impact of providing the First Steps program on the stories of people with Parkinson's Disease (PD) and to investigate the psychosocial and emotional mechanisms which may explain this impact. Methods: A qualitative study using a subtle realist paradigm and hermeneutic phenomenological methodology was undertaken. A single semi-structured interview was used to consider the impact and experiences of people with PD who completed either the intervention (2-day peer-led behavior intervention using storytelling 6-8 weeks apart) or received telephone support calls as part of the active control group. Descriptive statistics and a narrative analysis were undertaken on the results. Results: Forty-two participants were invited to participate, forty of whom completed the interview. This included 18 from the intervention group and 22 from the active control group. The intervention group identified the value of the program as worth-while, demonstrating improved exercise behavior and coping mechanisms following the intervention. Three major stories (the affirmed, the validated and the transformed story) identified the impact of the intervention. Three internal mechanisms (perceived control, hope and action, and the individual's mind set) alongside three social mechanisms (social comparison, social control and the first opportunity to share with peers) appeared to explain this impact. Conclusion: This study provides exciting and novel evidence of the impact of a peer-led psycho-educational intervention for people newly diagnosed with PD. Further research is needed to consider the impact of stories-based approaches on participants and consider a critical evaluation of the mechanisms which may explain changes in stories and self-reported behaviour.


Predicting disability progression and cognitive worsening in multiple sclerosis using patterns of grey matter volumes.
In multiple sclerosis (MS), MRI measures at the whole brain or regional level are only modestly associated with disability, while network-based measures are emerging as promising prognostic markers. We sought to demonstrate whether data-driven patterns of covarying regional grey matter (GM) volumes predict future disability in secondary progressive MS (SPMS). We used cross-sectional structural MRI, and baseline and longitudinal data of Expanded Disability Status Scale, Nine-Hole Peg Test (9HPT) and Symbol Digit Modalities Test (SDMT), from a clinical trial in 988 people with SPMS. We processed T1-weighted scans to obtain GM probability maps and applied spatial independent component analysis (ICA). We repeated ICA on 400 healthy controls. We used survival models to determine whether baseline patterns of covarying GM volume measures predict cognitive and motor worsening. We identified 15 patterns of regionally covarying GM features. Compared with whole brain GM, deep GM and lesion volumes, some ICA components correlated more closely with clinical outcomes. A mainly basal ganglia component had the highest correlations at baseline with the SDMT and was associated with cognitive worsening (HR=1.29, 95% CI 1.09 to 1.52, p<0.005). Two ICA components were associated with 9HPT worsening (HR=1.30, 95% CI 1.06 to 1.60, p<0.01 and HR=1.21, 95% CI 1.01 to 1.45, p<0.05). ICA measures could better predict SDMT and 9HPT worsening (C-index=0.69-0.71) compared with models including only whole and regional MRI measures (C-index=0.65-0.69, p value for all comparison <0.05). The disability progression was better predicted by some of the covarying GM regions patterns, than by single regional or whole-brain measures. ICA, which may represent structural brain networks, can be applied to clinical trials and may play a role in stratifying participants who have the most potential to show a treatment effect.


Multicenter validation of American Association for the Surgery of Trauma grading system for acute colonic diverticulitis and its use for emergency general surgery quality improvement program.
The American Association for the Surgery of Trauma (AAST) has developed a new grading system for uniform description of anatomic severity of emergency general surgery (EGS) diseases, ranging from Grade I (mild) to Grade V (severe). The purpose of this study was to determine the relationship of AAST grades for acute colonic diverticulitis with patient outcomes. A secondary purpose was to propose an EGS quality improvement program using risk-adjusted center outcomes, similar to National Surgical Quality Improvement Program and Trauma Quality Improvement Program methodologies. This was a retrospective study of 1,105 patients (one death) from 13 centers. At each center, two reviewers (blinded to each other's assignments) assigned AAST grades. Interrater reliability was measured using κ coefficient. Relationship between AAST grade and clinical events (complications, intensive care unit use, surgical intervention, and 30-day readmission) as well as length of stay was measured using regression analyses to control for age, comorbidities, and physiologic status at the time of admission. Final model was also used to calculate observed-to-expected (O-E) ratios for adverse outcomes (death, complications, readmissions) for each center. Median age was 54 years, 52% were males, 43% were minorities, and 22% required a surgical intervention. Almost two thirds had Grade I or II disease. There was a high level of agreement for grades between reviewers (κ = 0.81). Adverse events increased from 13% for Grade I, to 18% for Grade II, 28% for Grade III, 44% for Grade IV, and 50% for Grade V. Regression analysis showed that higher disease grades were independently associated with all clinical events and length of stay, after adjusting for age, comorbidities, and physiology. O-E ratios showed statistically insignificant variations in risk of death, complications, or readmissions. AAST grades for acute colonic diverticulitis are independently associated with clinical outcomes and resource use. EGS quality improvement program methodology that incorporates AAST grade, age, comorbidities, and physiologic status may be used for measuring quality of EGS care. High-quality EGS registries are essential for developing meaningful quality metrics. Prognostic study, level V.


Coadministration of an opioid agonist and antagonist for pain control.
The increased use of opioids in the treatment of chronic pain encourages the search for drugs with low abuse and tolerance potential but with potent analgesic activity. Opioid agonist-antagonists and partial agonists have less abuse potential than do mu opioid receptor agonists such as morphine, and have been used for many years for their analgesic affects. Recently they have been approved for treatment of opioid addiction. As a guard against abuse, an opioid antagonist, such as naloxone, is added to some opioid formulations. Doctors are often hesitant to prescribe agonist-antagonists and partial agonists to opioid-tolerant patients, fearing that these drugs may precipitate withdrawal. Can drugs being used safely for addiction treatment also safely replace opioid agonists to provide analgesia in chronic pain patients who are opioid-tolerant?


Amantadine and sparteine inhibit ATP-regulated K-currents in the insulin-secreting beta-cell line, HIT-T15.
1. The effects of pharmacological agents that potentiate insulin release were studied on ATP-regulated K-currents (K-ATP currents) in the insulin-secreting beta-cell line HIT-T15 by use of patch-clamp methods. 2. The tricyclic drug, 1-adamantanamine (amantadine), reversibly inhibited both whole-cell currents (with a Ki of 120 microM) and single channel currents in inside-out patches. This effect was principally due to an increase in a long closed state which reduced the channel open probability. The related compound, 1-adamantanol, in which the amino group is substituted by a hydroxyl one, did not inhibit K-ATP currents substantially. 3. The alkaloid, sparteine, reversibly inhibited both whole-cell K-ATP currents (Ki = 171 microM) and single channel currents in inside-out patches. 4. The results suggest that sparteine and amantadine can block the K-ATP channel from either side of the membrane and support the idea that at least part of the stimulatory effect of these agents on insulin secretion results from inhibition of this channel.


Risk and Emotion Among Healthy Volunteers in Clinical Trials.
Theorized as objective or constructed, risk is recognized as unequally distributed across social hierarchies. Yet the process by which social forces shape risk and risk emotions remains unknown. The pharmaceutical industry depends on healthy individuals to voluntarily test early-stage, investigational drugs in exchange for financial compensation. Emblematic of risk in late modernity, Phase I testing is a rich site for examining how class and race shape configurations of emotion and risk. Using interview data from 178 healthy trial participants, this article examines emotion and risk as mutually constituting processes linked to biographical context and social structure. Biographical events like economic insecurity and incarceration influence how risk is felt by providing comparative experiences of felt risk and felt benefits. Such events, in turn, are structured by class-based and racial inequalities, linking class and race positions to primary emotional experiences of risk.


Similarities between maternal and fetal RR interval tachograms and their association with fetal development.
An association between maternal and fetal heart rate (HR) has been reported but, so far, little is known about its physiological implication and importance relative to fetal development. Associations between both HRs were investigated previously by performing beat-by-beat coupling analysis and correlation analysis between average maternal and fetal HRs. However, studies reporting on the presence of similarities between maternal and fetal HRs or RR intervals (RRIs) over the short term (e.g., 5-min) at different gestational ages (GAs) are scarce. Here, we demonstrate the presence of similarities in the variations exhibited by maternal and fetal RRl tachograms (RRITs). To quantify the same similarities, a cross-correlation (CC) analysis between resampled maternal and fetal RRITs was conducted; RRITs were obtained from non-invasive electrocardiogram (ECG). The degree of similarity between maternal and fetal RRITs (bmfRRITs) was quantified by calculating four CC coefficients. CC analysis was performed for a total of 330 segments (two 5-min segments from 158 subjects and one 5-min from 14 subjects). To investigate the association of the similarity bmfRRITs with fetal development, the linear correlation between the calculated CC coefficients and GA was calculated. The results from the latter analysis showed that similarities bmfRRITs are common occurrences, they can be negative or positive, and they increase with GA suggesting the presence of a regulation that is associated with proper fetal development. To get an insight into the physiological mechanisms involved in the similarity bmfRRITs, the association of the same similarity with maternal and fetal HR variability (HRV) was investigated by comparing the means of two groups in which one of them had higher CC values compared to the other. The two groups were created by using the data from the 158 subjects where fetal RRI (fRRI) calculation from two 5-min ECG segments was feasible. The results of the comparison showed that the maternal very low frequency (VLF) HRV parameter is potentially associated with the similarity bmfRRITs implying that maternal hormones could be linked to the regulations involved in the similarity bmfRRITs. Our findings in this study reinforce the role of the maternal intrauterine environment on fetal development.


Frequency of public child care service use and associated factors.
Assess the maternal socio-demographic factors associated with the frequency of use of child care services by low income families. Cross-sectional analysis of 393 children between 12 and 16 months old who participated in a randomized field trial during their first year of life in a program of nutritional intervention. The study began in the Brazilian Unified Health System (Sistema Unico de Saúde, SUS) of the maternity hospital in the city of São Leopoldo, state of Rio Grande do Sul, Brazil. Children were examined regarding child care follow-up and vaccination schedule by checking the immunization chart. Data were analyzed using statistical tests Pearson's chi square and prevalence ratio (PR) with respective 95% confidence interval. The frequency of children who were not continuously taken to the child care service was 53.2%. Multivariate analysis suggests that the factors associated with the lack of continuous use of the service were: mother's educational level <or= 8 years (PR 1.32 95%CI 1.02-1.71), non-nuclear family structure (PR 1.32 95%CI 1.10-1.59) and not being an only child (PR 1.38 95%CI 1.10-1.72). The reasons for lack of follow-up, according to the mothers were: the fact that they thought it unnecessary for 66.2%, problems with the service for 21.7%, difficulties related to their jobs for 6.05%, and other reasons for 6.05%. The high frequency of children who were not taken to the child health care service for follow-up is associated with low maternal educational level and family structure, as well as the perception that follow-up visits are not necessary when the child does not have a disease.


Immediate Effects of Aquatic Therapy on Balance in Older Adults with Upper Limb Dysfunction: An Exploratory Study.
Aquatic physiotherapy has been shown to be effective in developing balance, strength, and functional reach over time. When dealing with immediate effects, the literature has concentrated more on the body's physiological response to the physical and mechanical properties of water during passive immersion. The purpose of this study was to evaluate the effects of a single 45-min active aquatic physiotherapy session on standing balance and strength, and its relationship with functional reach in persons 55 years and older with upper limb dysfunction. The intervention group (n = 12) was assessed before and after a single aquatic physiotherapy session, while the control group (n = 10) was evaluated before and after 45 min of sitting rest. Functional assessment was made using the visual analogue pain scale (points), step test (repetitions), functional reach test (cm), and global balance-standing test on a force platform (% time). A two-way repeated-measures ANOVA was applied (p < 0.05). The intervention group showed non-significant improvements between measurement before and after the intervention: Pain: 6.2 ± 1.9 vs. 5.2 ± 2.3 cm, steps: 7.0 ± 2.0 vs. 7.4 ± 1.8 repetitions, reach: 9.1 ± 2.8 vs. 10.4 ± 3.8 cm, and balance: 61.7 ± 5.9 vs. 71.3 ± 18.2% time in balance on the platform. The control group showed fewer changes but had better baseline values. A comparison between groups with time showed no significant differences in these changes. No significant immediate effects were found for one session of aquatic physiotherapy applied to patients older than 55 years with upper limb dysfunction.


Microbiological quality and safety of Rastrineobola argentea retailed in Kisumu town markets, Kenya.
To investigate faecal contamination and safety of Rastrineobola argentea sold in retail markets in Kisumu town. This was a repeated cross sectional study and based on random sampling. Kisumu city, targeting six markets; Oile, Jubilee, Kibuye, Kondele, Nyalenda and Manyatta. A total of 60 fish samples were analysed. All the fish were found to be contaminated with E. coli, and in addition 6.67% of the fish products tested positive for Salmonella. Shigella was absent in all samples analysed. 26.53% of E. coli isolates tested were resistant to two or more antimicrobial agents tested, with the highest level of resistance detected against cotrimoxazole at 38.76%. The E. coli multiple antibiotic resistance (MAR) index was 0.084 indicating that the contamination was not originating from a high-risk source. A plasmid of approximately 5.6 kb was commonly isolated from E. coli isolates that showed resistance to ampicillin. Plasmids isolated were not transferable by conjugation. The presence of Salmonella spp and occurrence of MDR E. coli were identified as some of the possible health risks that may be associated with R. argentea displayed for sale in Kisumu city markets. This possess a real health risk through consumption or directly through contact with the fish products.