On the mechanism of antidepressant-like action of berberine chloride.
Berberine, an alkaloid isolated from Berberis aristata Linn. has been used in the Indian system of medicines as a stomachic, bitter tonic, antiamoebic and also in the treatment of oriental sores. Evidences have demonstrated that berberine possesses central nervous system activities, particularly the ability to inhibit monoamine oxidase-A, an enzyme involved in the degradation of norepinephrine and serotonin (5-HT). With this background, the present study was carried out to elucidate the antidepressant-like effect of berberine chloride in different behavioural paradigms of despair. Berberine (5, 10, 20 mg/kg, i.p.) inhibited the immobility period in mice in both forced swim and tail-suspension test, however, the effect was not dose-dependent. Berberine (5 and 10 mg/kg, i.p.) also reversed the reserpine-induced behavioral despair. Berberine (5 mg/kg, i.p.) enhanced the anti-immobility effect of subeffective doses of various typical but not atypical antidepressant drugs in forced swim test. Berberine (5 mg/kg, i.p.) following its acute administration in mice resulted in increased levels of norepinephrine (31%), serotonin (47%) and dopamine (31%) in the whole brain. Chronic administration of berberine (5 mg/kg, i.p.) for 15 days significantly increased the levels of norepinephrine (29%), serotonin (19%) as well as dopamine (52%) but at higher dose (10 mg/kg, i.p.), there was no change in the norepinephrine (12%) levels but a significant increase in the serotonin (53%) and dopamine (31%) levels was found. The antidepressant-like effect of berberine (5 mg/kg, i.p.) in forced swim test was prevented by pretreatment with l-arginine (750 mg/kg, i.p.) or sildenafil (5 mg/kg, i.p.). On the contrary, pretreatment of mice with 7-nitroindazole (7-NI) (25 mg/kg, i.p.) or methylene blue (10 mg/kg, i.p.) potentiated the effect of berberine (2 mg/kg, i.p.) in the forced swim test. Pretreatment of mice with (+)-pentazocine (2.5 mg/kg, i.p.), a high-affinity sigma1 receptor agonist, produced synergism with subeffective dose of berberine (2 mg/kg, i.p.). Pretreatment with various sigma receptor antagonists viz. progesterone (10 mg/kg, s.c.), rimcazole (5 mg/kg, i.p.) and N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(dimethylamino)ethylamine (BD1047; 1 mg/kg, i.p.) reversed the anti-immobility effects of berberine (5 mg/kg, i.p.). Berberine at lower dose did not affect the locomotor activity and barbiturate-induced sleep time. It produced mild hypothermic action in rats and displayed analgesic effect in mice. Taken together, theses findings demonstrate that berberine exerted antidepressant-like effect in various behavioural paradigms of despair possibly by modulating brain biogenic amines (norepinephrine, serotonin and dopamine). Further, nitric oxide pathway and/or sigma receptors are involved in mediating its antidepressant-like activity in mouse forced swim test.


Negative oncologic impact of poor postoperative pain control in left-sided pancreatic cancer.
To investigate the association between postoperative pain control and oncologic outcomes in resected pancreatic ductal adenocarcinoma (PDAC). From January 2009 to December 2014, 221 patients were diagnosed with PDAC and underwent resection with curative intent. Retrospective review of the patients was performed based on electronic medical records system. One patient without records of numerical rating scale (NRS) pain intensity scores was excluded and eight patients who underwent total pancreatectomy were also excluded. NRS scores during 7 postoperative days following resection of PDAC were reviewed along with clinicopathologic characteristics. Patients were stratified into a good pain control group and a poor pain control group according to the difference in average pain intensity between the early (POD 1, 2, 3) and late (POD 5, 7) postoperative periods. Cox-proportional hazards multivariate analysis was performed to determine association between postoperative pain control and oncologic outcomes. A total of 212 patients were dichotomized into good pain control group (n = 162) and poor pain control group (n = 66). Median follow-up period was 17 mo. A negative impact of poor postoperative pain control on overall survival (OS) was observed in the group of patients receiving distal pancreatectomy (DP group; 42.0 mo vs 5.0 mo, P = 0.001). Poor postoperative pain control was also associated with poor disease-free survival (DFS) in the DP group (18.0 mo vs 8.0 mo, P = 0.001). Patients undergoing pancreaticoduodenectomy or pylorus-preserving pancreaticoduodenectomy (PD group) did not show associations between postoperative pain control and oncologic outcomes. Poor patients' perceived pain control was revealed as an independent risk factor of both DFS (HR = 4.157; 95%CI: 1.938-8.915; P < 0.001) and OS (HR = 4.741; 95%CI: 2.214-10.153; P < 0.001) in resected left-sided pancreatic cancer. Adequate postoperative pain relief during the early postoperative period has important clinical implications for oncologic outcomes after resection of left-sided pancreatic cancer.


The origin of the spinal subdural space: ultrastructure findings.
Previous studies of samples from cranial meninges have created doubts about the existence of a virtual subdural space. We examined the ultrastructure of spinal meninges from three human cadavers immediately after death to see whether there is a virtual subdural space at this level. The arachnoid mater had two portions: a compact laminar portion covering the dural sac internal surface and a trabecular portion extending like a spider web around the pia mater. There was a cellular interface between the laminar arachnoid and the internal layer of the dura that we called the dura-arachnoid interface. There was no subdural space in those specimens where the dura mater was macroscopically in continuity with the arachnoid trabecules. In the specimens where the dura mater was separated from the arachnoid, we found fissures in between the neurothelial cells that extended throughout the interface. We hypothesize that the subdural space would have its origin within the dura-arachnoid interface when the neurothelial cells break up, creating in this way a real subdural space. The subdural space was not seen under transmission electron microscopy in samples of human spinal meninges where surgical manipulation was avoided. Scanning electron microscopy in other samples showed the presence of broken neurothelial cells giving up fissures that extended along the dura-arachnoid interface. These findings may explain the origin of a real subdural space.


The Effect of Genetically Guided Mathematical Prediction and the Blood Pressure Response to Pharmacotherapy in Hypertension Patients.
The purpose of this study was to determine the effectiveness of a simple algorithm to mathematically predict a patients' response to blood pressure (BP) therapy using functional genes in the 3 major organ systems involved in hypertension. Eighty-six patients with controlled hypertension completed 1 study visit consisting of a buccal swab collection, measurement of office BP, and a medical chart review for BP history. Genes in the analysis included 14 functional alleles in 11 genes. These genotypes were mathematically summed per organ system to determine whether a patient would likely respond to target therapy. Patients recommended to and taking a diuretic had significantly higher rates of control (<120/<80) than patients recommended but not taking this drug class (0.2 ± 0.1 and 0.03 ± 0.03, respectively). Furthermore, there was a difference between patients genetically recommended and taking an angiotensin receptor blocker (ARB) vs patients recommended but not taking an ARB for the lowest diastolic blood pressure (DBP) and mean arterial pressure (MAP) recorded in the past 2 years (DBP = 66.2 ± 2.9 and 75.3 ± 1.7, MAP = 82.3 ± 2.8 and 89.3 ± 1.5, respectively). In addition, there was a nonsignificant trend for greater reductions in ΔSBP, ΔDBP, and ΔMAP in patients on recommended drug class for beta-blockers, diuretics, and angiotensin II receptor blockers vs patients not on these classes. The present study suggests that simple mathematical weighting of functional genotypes known to control BP may be ineffective in predicting control. This study demonstrates the need for a more complex, weighted, multigene algorithm to more accurately predict BP therapy response.


Single-molecule spectroscopy reveals chaperone-mediated expansion of substrate protein.
Molecular chaperones are an essential part of the machinery that avoids protein aggregation and misfolding in vivo. However, understanding the molecular basis of how chaperones prevent such undesirable interactions requires the conformational changes within substrate proteins to be probed during chaperone action. Here we use single-molecule fluorescence spectroscopy to investigate how the DnaJ-DnaK chaperone system alters the conformational distribution of the denatured substrate protein rhodanese. We find that in a first step the ATP-independent binding of DnaJ to denatured rhodanese results in a compact denatured ensemble of the substrate protein. The following ATP-dependent binding of multiple DnaK molecules, however, leads to a surprisingly large expansion of denatured rhodanese. Molecular simulations indicate that hard-core repulsion between the multiple DnaK molecules provides the underlying mechanism for disrupting even strong interactions within the substrate protein and preparing it for processing by downstream chaperone systems.


Protein intake from 0 to 18 years of age and its relation to health: a systematic literature review for the 5th Nordic Nutrition Recommendations.
The present systematic literature review is a part of the 5th revision of the Nordic Nutrition Recommendations. The aim was to assess the health effects of different levels of protein intake in infancy and childhood in a Nordic setting. The initial literature search resulted in 435 abstracts, and 219 papers were identified as potentially relevant. Full paper selection resulted in 37 quality-assessed papers (4A, 30B, and 3C). A complementary search found four additional papers (all graded B). The evidence was classified as convincing, probable, limited-suggestive, and limited-inconclusive. Higher protein intake in infancy and early childhood is convincingly associated with increased growth and higher body mass index in childhood. The first 2 years of life is likely most sensitive to high protein intake. Protein intake between 15 E% and 20 E% in early childhood has been associated with an increased risk of being overweight later in life, but the exact level of protein intake above which there is an increased risk for being overweight later in life is yet to be established. Increased intake of animal protein in childhood is probably related to earlier puberty. There was limited-suggestive evidence that intake of animal protein, especially from dairy, has a stronger association with growth than vegetable protein. The evidence was limited-suggestive for a positive association between total protein intake and bone mineral content and/or other bone variables in childhood and adolescence. Regarding other outcomes, there were too few published studies to enable any conclusions. In conclusion, the intake of protein among children in the Nordic countries is high and may contribute to increased risk of later obesity. The upper level of a healthy intake is yet to be firmly established. In the meantime, we suggest a mean intake of 15 E% as an upper limit of recommended intake at 12 months, as a higher intake may contribute to increased risk for later obesity.


The effects of pantethine on lipid and lipoprotein abnormalities in survivors of cerebral infarction.
A daily 1000 mg of pantethine was orally administered to 12 male survivors of cerebral infarction for 3 months. Before and after the medication, plasma lipoproteins were ultracentrifugally separated. Plasma concentrations of total cholesterol, triglyceride, phospholipid, and VLDL- and LDL-cholesterol tended to decrease. On the contrary, HDL-cholesterol concentration and HDL:LDL-cholesterol ratio significantly increased. HDL2-cholesterol concentration significantly increased, whereas HDL3-cholesterol concentration remained unchanged, resulting in a significant elevation of HDL2:HDL3-cholesterol ratio. In HDL2 the percentage content of cholesterol and triglyceride decreased, while that of phospholipid increased. However, no significant changes were found in the chemical composition of HDL3. The plasma concentrations of apo A-I and, to a lesser extent, A-II increased, with a slight increase in A-I:A-II ratio but A-I:A-II ratio in either HDL2 or HDL3 remained unchanged.


Online image guided tumour tracking with scanned proton beams: a comprehensive simulation study.
Tumour tracking with scanned particle beams potentially requires accurate 3D information on both tumour motion and related density variations. We have previously developed a model-based motion reconstruction method, which allows for the prediction of deformable motions from sparsely sampled surrogate motions tracked via an on-board imaging system (Zhang et al (2013 Phys. Med. Biol. 58 8621)). Here, we investigate the potential effectiveness of tumour tracking for scanned proton beam therapy using such an approach to guide scanned beam tracking, together with the effectiveness of 're-tracking' for reducing residual motion effects due to tracking uncertainties. Three different beam tracking strategies (2D, 2D deformable and 3D) have been applied to three different liver motion cases, with mean magnitudes ranging from 10-20 mm. All simulations have been performed using simulated 4DCTs derived from 4DMRI datasets, whereby inter-breath-cycle motion variability is taken into account. The results show that, without beam tracking, large interplay effects are observed for all motion cases, resulting in CTV D5-95 values of 34.9/58.5/79.4% for the three cases, respectively. These can be reduced to 16.9/18.8/29.1% with 2D tracking, to 15.5/17.9/23.3% with 2D deformable tracking and to 15.1/17.8/21.0% with 3D tracking. Clear 'inverse interplay' effects have also been observed in the proximal portion of the field. However, with three-times re-tracking, D5-95 for the largest motions (20 mm) can be reduced to 13.0/12.8% for 2D and 3D tracking, respectively, and 'hot spots' resulting from the 'inverse interplay' effect can be substantially reduced. In summary, we have found that, for motions over 10 mm, tracking alone cannot fully mitigate motion effects, and can lead to substantially increased doses to normal tissues in the entrance path of the field. However, three-times re-tracking substantially improves the effectiveness of all types of beam tracking, with substantial advantages of 3D over 2D re-tracking only being observed for the largest motion scenario investigated.


Formation of Tap/NXT1 heterodimers activates Tap-dependent nuclear mRNA export by enhancing recruitment to nuclear pore complexes.
The Tap protein has been shown to activate the nuclear export of mRNA species bearing retroviral constitutive transport elements and is also believed to play an essential role in the sequence nonspecific export of cellular mRNAs. However, it has remained unclear how Tap activity is regulated in vivo. Here, we report that the small NXT1/p15-1 protein functions as a critical cofactor for Tap-mediated mRNA export in both human and invertebrate cells. In the absence of NXT1 binding, the Tap protein is unable to effectively interact with components of the nuclear pore complex and both Tap nucleocytoplasmic shuttling and the nuclear export of mRNA molecules tethered to Tap are therefore severely attenuated. Formation of a Tap/NXT1 heterodimer enhances nucleoporin binding both in vitro and in vivo and induces the formation of a Tap/NXT1/nucleoporin ternary complex that is likely to be a key intermediate in the process of nuclear mRNA export. The critical importance of NXT1 for the nuclear export of poly(A)(+) RNA is emphasized by the finding that specific inhibition of the expression of the Drosophila homolog of human NXT1, by using RNA interference, results in the nuclear accumulation of poly(A)(+) RNA in cultured insect cells. These data suggest that NXT1 may act as a molecular switch that regulates the ability of Tap to mediate nuclear mRNA export by controlling the interaction of Tap with components of the nuclear pore.


Listeria monocytogenes meningitis in an immunocompromised patient.
This report describes a case of meningitis caused by Listeria monocytogenes in a stem cell transplant recipient on immunosuppressive therapy for cutaneous chronic graft-versus host disease. A 59-year-old woman had undergone allogeneic stem cell transplantation (from a matched unrelated donor) 13 months previously for chronic lymphocytic leukemia. She was on regular hematologic follow-up. Though her previous malignancy has been in remission, she was immunosuppressed due to the pharmacological treatment. We describe a meningitis caused by a typical food-borne pathogen, dangerous in patients with impaired cell-mediated immunity. Moreover the bacterium had a multidrug resistance, a rare characteristic in clinical listeriosis. Rapid diagnosis and treatment are key factors in these cases. We chose ampicillin and rifampicin that allowed a complete resolution of the clinical manifestations.


Effects of acupuncture stimulation on recovery ability of male elite basketball athletes.
Developing effective methods for helping athletes recover from muscle fatigue is deemed essential, particularly on the eves' important competitions. This study aimed to investigate the effects of acupuncture stimulation on athletes' recovery abilities. Subjects were selected from 30 male elite university basketball players who were randomly assigned to 3 groups: acupuncture group, sham group, and normal (control) group, each containing 10 subjects. Acupuncture was carried out on each athlete in acupuncture group at the Neiguan (PC6) and Zusanli (ST36) acupoints, beginning at 15 min prior to exercise and continuing until exhaustion of the subject. Similar acupuncture was also carried out on each athlete in the sham group but the positions were 1 cm away from the PC6 and ST36 acupoints. No acupuncture was conducted on the athletes in the normal group. During the experiments, each subject performed separate runs on the treadmill. The data of heart rate (HR(max)), oxygen consumption (VO(2max)), and blood lactic acid were measured during the rest period and at 3 recovery points of time (5th, 30th and 60th min) post-exercise. The results showed that the acupuncture group (PC6 and ST36) has significantly lower HR(max), VO(2max) and blood lactic acid than both the sham and normal groups at the 30th min post-exercise. Blood lactic acid of the acupuncture group was also significantly lower than that of the other two groups in the 60th min post-exercise. Our findings have shed some light on the development of effective acupuncture schemes to enhance the recovery ability for elite basketball athletes.


Smad4 loss is associated with fewer S100A8-positive monocytes in colorectal tumors and attenuated response to S100A8 in colorectal and pancreatic cancer cells.
S100A8 and its dimerization partner S100A9 are emerging as important chemokines in cancer. We previously reported that Smad4-negative pancreatic tumors contain fewer stromal S100A8-positive monocytes than their Smad4-positive counterparts. Here, we studied S100A8/A9-expressing cells in colorectal tumors relating their presence to clinicopathological parameters and Smad4 status. Two-dimensional gel electrophoresis (n = 12) revealed variation in the levels of S100A8 protein in colorectal cancer tumors, whereas immunohistochemical analysis (n = 313) showed variation in the numbers of stromal S100A8-positive and S100A9-positive cells. Loss of Smad4 expression was observed in 42/304 (14%) colorectal tumors and was associated with reduced numbers of S100A8-positive (P = 0.03) but not S100A9-positive stromal cells (P = 0.26). High S100A9 cell counts were associated with large tumor sizes (P = 0.0006) and poor differentiation grade (P = 0.036). However, neither S100A8 nor S100A9 cell counts predicted poor survival, except for patients with Smad4-negative tumors (P = 0.02). To address the impact of environmental S100A8/A9 chemokines on tumor cells, we examined the effects of exogenously added S100A8 and S100A9 proteins on cellular migration and proliferation of colorectal and pancreatic cancer cells. S100A8 and S100A9 enhanced migration and proliferation in Smad4-positive and Smad4-negative cancer cells. However, transient depletion of Smad4 resulted in loss of responsiveness to exogenous S100A8, but not S100A9. S100A8 and S100A9 activated Smad4 signaling as evidenced by phosphorylation of Smad2/3; blockade of the receptor for the advanced glycation end products inhibited this response. In conclusion, Smad4 loss alters the tumor's interaction with stromal myeloid cells and the tumor cells' response to the stromal chemokine, S100A8.


Chemo-photodynamic combined gene therapy and dual-modal cancer imaging achieved by pH-responsive alginate/chitosan multilayer-modified magnetic mesoporous silica nanocomposites.
Multifunctional theranostics have offered some interesting new opportunities for cancer therapy and diagnosis in the last decade. Herein, magnetic mesoporous silica nanoparticles (M-MSNs) were designed and synthesized, then the photosensitizer chlorin e6 (Ce6) and antitumor drug doxorubicin (Dox) were adsorbed onto the M-MSNs. Biocompatible alginate/chitosan polyelectrolyte multilayers (PEM) were assembled on the M-MSNs to achieve a pH-responsive drug delivery system and adsorb P-gp shRNA for reversing the multidrug resistance. The obtained M-MSN(Dox/Ce6)/PEM/P-gp shRNA nanocomposites were characterized using TEM, SEM, X-ray diffraction, BET, FTIR and electrophoresis. The nanocomposites with average diameter of 280 nm exhibited a pH-responsive drug release profile, and more singlet oxygen generation in cancer cells after laser illumination. CCK-8 assay and calcein-AM/PI co-staining showed that the multifunctional nanocomplexes significantly increased cell apoptosis in vitro. With tumor-bearing Balb/c mice employed as the animal model, combined photodynamic therapy and chemotherapy was carried out, also achieving synergistic anti-tumor effects in vivo. The cores of bifunctional Fe3O4-Au nanoparticles in the multifunctional nanocomposites enabled dual-modal MR and CT imaging, which illustrated strong tumor uptake of these nanocomposites after intravenous injection into tumor-bearing mice. This work highlights the great potential of magnetic mesoporous silica nanocomposites as a multifunctional delivery platform, which is promising for imaging-guided cancer combination therapy with high efficacy.


Effect of bleaching on fracture toughness of composite-dentin bonds.
The effect of carbamide peroxide concentration and length of exposure on fracture toughness (KIC) of existing composite-dentin interfaces was assessed using the notchless triangular prism (NTP) specimen KIC test. Freshly extracted human molars and premolars were wet ground on 600-grit SiC to obtain 4 x 4 x 4 x 4 mm triangular prisms, exposing buccal or lingual dentin. Dentin surfaces were bonded using a resin composite (Z-250, 3M) and SingleBond dentin bonding system (3M) to obtain 180 8-mm-long dentin-composite NTP specimens. The bonding system was applied using the "wet-bonding" technique. The bonded specimens were randomly assigned to 20 groups. The effect of exposure to three control solutions (tap water, Carbopol, and Carbopol-urea) and to four concentrations (11%, 13%, 16%, 21%) of carbamide peroxide (Perfecta, Premier) was assessed. Testing was conducted after 1, 2, and 3 weeks representing a cumulative exposure of 14, 42, and 70 h. The maximum force recorded before fracture was used to calculate KIC. The data were statistically analysed using two-factor ANOVA followed by one-way ANOVA and Bonferroni multiple means comparisons. Selected fractured surfaces were characterized using SEM. Data analysis revealed that cumulative exposure to bleaching agent for 70 h significantly (p < 0.01) decreased the interfacial KIC, regardless of concentration. For the 16% and 21% concentrations, a significant reduction was observed after 42 h. The results suggest that bleaching could adversely affect the interfacial fracture toughness of dentin-resin composite adhesive interfaces.


A game theoretic framework for analyzing re-identification risk.
Given the potential wealth of insights in personal data the big databases can provide, many organizations aim to share data while protecting privacy by sharing de-identified data, but are concerned because various demonstrations show such data can be re-identified. Yet these investigations focus on how attacks can be perpetrated, not the likelihood they will be realized. This paper introduces a game theoretic framework that enables a publisher to balance re-identification risk with the value of sharing data, leveraging a natural assumption that a recipient only attempts re-identification if its potential gains outweigh the costs. We apply the framework to a real case study, where the value of the data to the publisher is the actual grant funding dollar amounts from a national sponsor and the re-identification gain of the recipient is the fine paid to a regulator for violation of federal privacy rules. There are three notable findings: 1) it is possible to achieve zero risk, in that the recipient never gains from re-identification, while sharing almost as much data as the optimal solution that allows for a small amount of risk; 2) the zero-risk solution enables sharing much more data than a commonly invoked de-identification policy of the U.S. Health Insurance Portability and Accountability Act (HIPAA); and 3) a sensitivity analysis demonstrates these findings are robust to order-of-magnitude changes in player losses and gains. In combination, these findings provide support that such a framework can enable pragmatic policy decisions about de-identified data sharing.


Effects of dry needling in HIP muscles in patients with HIP osteoarthritis: A randomized controlled trial.
Dry needling (DN) in active myofascial trigger points (MTrPs) is effective to reduce pain, increase range of motion (ROM) and improve physical function in different musculoskeletal disorders. However, there is a lack of studies evaluating the effects of DN in active MTrPs in hip osteoarthritis (OA). To determine the short-term effects of DN on pain, hip ROM and physical function in patients with hip OA. Double-blind randomized controlled trial. Thirty patients with unilateral hip OA were randomized into two groups: DN group and sham group. Participants received three treatment sessions. The treatment was applied in active MTrPs of the iliopsoas, rectus femoris, tensor fasciae latae and gluteus minimus muscles. Pain intensity (visual analogic scale), passive hip ROM (universal goniometer and digital inclinometer) and physical function (30s chair-stand test and 20m walk test) were assessed at baseline and after the three treatment sessions. There was decreased pain intensity, increased hip ROM, and improved physical function following the DN treatment. These improvements were statistically significant (p < 0.05) compared to the sham group. The sham group had increased pain intensity and decreased hip ROM (p < 0.05). Pain, hip ROM, and physical function improved after the application of DN in active MTrPs of the hip muscles in patients with hip OA.


Phylogeny of parasitic parabasalia and free-living relatives inferred from conventional markers vs. Rpb1, a single-copy gene.
Parabasalia are single-celled eukaryotes (protists) that are mainly comprised of endosymbionts of termites and wood roaches, intestinal commensals, human or veterinary parasites, and free-living species. Phylogenetic comparisons of parabasalids are typically based upon morphological characters and 18S ribosomal RNA gene sequence data (rDNA), while biochemical or molecular studies of parabasalids are limited to a few axenically cultivable parasites. These previous analyses and other studies based on PCR amplification of duplicated protein-coding genes are unable to fully resolve the evolutionary relationships of parabasalids. As a result, genetic studies of Parabasalia lag behind other organisms. Comparing parabasalid EF1α, α-tubulin, enolase and MDH protein-coding genes with information from the Trichomonas vaginalis genome reveals difficulty in resolving the history of species or isolates apart from duplicated genes. A conserved single-copy gene encodes the largest subunit of RNA polymerase II (Rpb1) in T. vaginalis and other eukaryotes. Here we directly sequenced Rpb1 degenerate PCR products from 10 parabasalid genera, including several T. vaginalis isolates and avian isolates, and compared these data by phylogenetic analyses. Rpb1 genes from parabasalids, diplomonads, Parabodo, Diplonema and Percolomonas were all intronless, unlike intron-rich homologs in Naegleria, Jakoba and Malawimonas. The phylogeny of Rpb1 from parasitic and free-living parabasalids, and conserved Rpb1 insertions, support Trichomonadea, Tritrichomonadea, and Hypotrichomonadea as monophyletic groups. These results are consistent with prior analyses of rDNA and GAPDH sequences and ultrastructural data. The Rpb1 phylogenetic tree also resolves species- and isolate-level relationships. These findings, together with the relative ease of Rpb1 isolation, make it an attractive tool for evaluating more extensive relationships within Parabasalia.


Histopathological examinations of rat brains after long-term exposure to GSM-900 mobile phone radiation.
In order to mimic the real life situation, with often life-long exposure to the electromagnetic fields emitted by mobile phones, we have investigated in a rat model the effects of repeated exposures under a long period to Global System for Mobile Communication-900 MHz (GSM-900) radiation. Out of a total of 56 rats, 32 were exposed once weekly in a 2-h period, for totally 55 weeks, at different average whole-body specific absorption rates (SAR) (of in average 0.6 and 60 mW/kg at the initiation of the experimental period). The animals were exposed in a transverse electromagnetic transmission line chamber (TEM-cell) to radiation emitted by a GSM-900 test phone. Sixteen animals were sham exposed and eight animals were cage controls, which never left the animal house. After behavioural tests, 5-7 weeks after the last exposure, the brains were evaluated for histopathological alterations such as albumin extravasation, dark neurons, lipofuscin aggregation and signs of cytoskeletal and neuritic neuronal changes of the type seen in human ageing. In this study, no significant alteration of any these histopathological parameters was found, when comparing the GSM exposed animals to the sham exposed controls.


Engineered tissue scaffolds with variational porous architecture.
This paper presents a novel computer-aided modeling of 3D tissue scaffolds with a controlled internal architecture. The complex internal architecture of scaffolds is biomimetically modeled with controlled micro-architecture to satisfy different and sometimes conflicting functional requirements. A functionally gradient porosity function is used to vary the porosity of the designed scaffolds spatially to mimic the functionality of tissues or organs. The three-dimensional porous structures of the scaffold are geometrically partition into functionally uniform porosity regions with a novel offsetting operation technique described in this paper. After determining the functionally uniform porous regions, an optimized deposition-path planning is presented to generate the variational internal porosity architecture with enhanced control of interconnected channel networks and continuous filament deposition. The presented methods are implemented, and illustrative examples are presented in this paper. Moreover, a sample optimized tool path for each example is fabricated layer-by-layer using a micronozzle biomaterial deposition system.


Effect of host change on demographic fitness of the parasitoid, Trichogramma brassicae.
Trichogramma brassicae (Bezdenko) is the most important species of Trichogramma parasitoids in Iran. The cotton bollworm, Helicoverpa armigera (Hubner) is a polyphagous insect pest that attacks many crops including cotton, maize, soybean, tomato, etc. The bollworm egg is a suitable target for many Trichogramma species. Factitious hosts such as eggs of the flour moth, Anagasta kuehniella (Zeller) and cereal moth, Sitotroga cerealella (Hubner) are used for mass rearing purposes. But a problem that arises sometimes in laboratory cultures is the development of a tendency toward laboratory hosts following a few generations rearing with them. This may tend to a low efficiency on target pest in field conditions. In this study the possibility of declining efficiency of the parasitoid on target pest by developing such a preference to alternative hosts in previous generations were investigated when the flour moth or cereal moth uses as laboratory host. Two generations of T. brassicae were reared on each of the mentioned hosts and then transferred to H. armigera eggs for two further generations. The intrinsic rate of natural increase as well as other life table parameters were used for monitoring fitness of the parasitoid at successive generations. Even generations were included to determine if previously rearing host affected parasitoid performance. Results revealed that host shift from cereal moth to bollworm caused a sudden fall in population growth parameters (both intrinsic rate of natural increase and net replacement rate). Further rearing on bollworm eggs led to a relapse in both parameters. No similar effect was observed in cultures initiated with the flour moth. As a whole, cereal moth was a more suitable host than flour moth.


Practices used for recommending sickness certification by general practitioners: a conversation analytic study of UK primary care consultations.
Existing research indicates that many patients and doctors find the process of negotiating sickness certification for time off work to be a difficult one. This study examined how patients and general practitioners (GPs) managed these negotiations in a sample of UK primary care consultations. The study made use of an existing dataset of audio-recorded consultations between 13 GPs and 506 unselected adult patients in five general practices in London. Forty-nine consultations included discussions for both initial and repeat sickness certification across a wide range of conditions. Here we report our findings on doctor practices for recommending, as opposed to patient practices for advocating for, sickness certification (n = 26 cases). All cases were transcribed in detail and analysed using conversation analytic methods. Four main communication practices were observed: (1) declarative statements of need for sickness certification; (2) 'do you need' offers for sickness certification; (3) 'do you want' offers for sickness certification; and (4) conditional 'If X, Y' offers for sickness certification. These different communication practices indexed doctor agency, doctor endorsement and patient entitlement to varying degrees. In the main, recommendations to patients presenting with biomedical problems or a repeat occurrence of a psychosocial problem displayed stronger doctor endorsement and patient entitlement. Contrastingly, recommendations to patients presenting with new psychosocial and biopsychosocial problems, displayed weaker endorsement and patient entitlement. This study offers new evidence to support the Parsonian argument that becoming sick involves entering a social role with special rights and obligations. Through documenting doctors' orientations to their gatekeeping role as well as patients' orientations to differential rights vis à vis legitimacy, we demonstrate the contrasting stances of doctors in situ when giving sick notes for biomedical problems as opposed to difficulties of a more psychosocial nature.


Glc7/protein phosphatase 1 regulatory subunits can oppose the Ipl1/aurora protein kinase by redistributing Glc7.
Faithful chromosome segregation depends on the opposing activities of the budding yeast Glc7/PP1 protein phosphatase and Ipl1/Aurora protein kinase. We explored the relationship between Glc7 and Ipl1 and found that the phosphorylation of the Ipl1 substrate, Dam1, was altered by decreased Glc7 activity, whereas Ipl1 levels, localization, and kinase activity were not. These data strongly suggest that Glc7 ensures accurate chromosome segregation by dephosphorylating Ipl1 targets rather than regulating the Ipl1 kinase. To identify potential Glc7 and Ipl1 substrates, we isolated ipl1-321 dosage suppressors. Seven genes (SDS22, BUD14, GIP3, GIP4, SOL1, SOL2, and PEX31) encode newly identified ipl1 dosage suppressors, and all 10 suppressors encode proteins that physically interact with Glc7. The overexpression of the Gip3 and Gip4 suppressors altered Glc7 localization, indicating they are previously unidentified Glc7 regulatory subunits. In addition, the overexpression of Gip3 and Gip4 from the galactose promoter restored Dam1 phosphorylation in ipl1-321 mutant cells and caused wild-type cells to arrest in metaphase with unsegregated chromosomes, suggesting that Gip3 and Gip4 overexpression impairs Glc7's mitotic functions. We therefore propose that the overexpression of Glc7 regulatory subunits can titrate Glc7 away from relevant Ipl1 targets and thereby suppress ipl1-321 cells by restoring the balance of phosphatase/kinase activity.


Culture, distress, and oxytocin receptor polymorphism (OXTR) interact to influence emotional support seeking.
Research has demonstrated that certain genotypes are expressed in different forms, depending on input from the social environment. To examine sensitivity to cultural norms regarding emotional support seeking as a type of social environment, we explored the behavioral expression of oxytocin receptor polymorphism (OXTR) rs53576, a gene previously related to socio-emotional sensitivity. Seeking emotional support in times of distress is normative in American culture but not in Korean culture. Consequently, we predicted a three-way interaction of culture, distress, and OXTR genotype on emotional support seeking. Korean and American participants (n = 274) completed assessments of psychological distress and emotional support seeking and were genotyped for OXTR. We found the predicted three-way interaction: among distressed American participants, those with the GG/AG genotypes reported seeking more emotional social support, compared with those with the AA genotype, whereas Korean participants did not differ significantly by genotype; under conditions of low distress, OXTR groups did not differ significantly in either cultural group. These findings suggest that OXTR rs53576 is sensitive to input from the social environment, specifically cultural norms regarding emotional social support seeking. These findings also indicate that psychological distress and culture are important moderators that shape behavioral outcomes associated with OXTR genotypes.


Efficacy of antidepressants in juvenile depression: meta-analysis.
The safety of antidepressants in children and adolescents is being questioned and the efficacy of these drugs in juvenile depression remains uncertain. To assess antidepressant efficacy in juvenile depression. Systematic review and meta-analysis of randomised controlled trials (RCTs) comparing responses to antidepressants, overall and by type, v. placebo in young people with depression. Thirty drug-placebo contrasts in RCTs lasting 8 weeks (median) involved 3069 participants (512 person-years) of average age 13.5 years. Meta-analysis yielded a modest pooled drug/placebo response rate ratio (RR=1.22, 95% CI 1.15-1.31), with little separation between antidepressant types. Findings were similar for response rate differences and corresponding number needed to treat (NNT): overall NNT=9; tricyclic antidepressants NNT=14 > serotonin reuptake inhibitors NNT=9 > other antidepressants NNT=8. Numbers needed to treat decreased with increasing age: children (NNT=21) > mixed ages (NNT=10) > adolescents (NNT=8). Antidepressants of all types showed limited efficacy in juvenile depression, but fluoxetine might be more effective, especially in adolescents. Studies in children and in severely depressed, hospitalised or suicidal juvenile patients are needed, and effective, safe and readily accessible treatments for juvenile depression are urgently required.


RE1 silencing transcription factor/neuron-restrictive silencing factor regulates expansion of adult mouse subventricular zone-derived neural stem/progenitor cells in vitro.
Adult neural stem cell (aNSC) activity is tuned by external stimuli through the recruitment of transcription factors. This study examines the RE1 silencing transcription factor (REST) in neural stem/progenitor cells isolated from the subventricular zone of adult mouse brain and provides the first extensive characterization of REST-mediated control of the cellular and molecular properties. This study shows that REST knockdown affects the capacity of progenitor cells to generate neurospheres, reduces cell proliferation, and triggers cell differentiation despite the presence of growth factors. Genome- and transcriptome-wide analyses show that REST binding sites are significantly enriched in genes associated with synaptic transmission and nervous system development and function. Seeking candidate regulators of aNSC function, this study identifies a member of the bone morphogenetic protein (BMP) family, BMP6, the mRNA and protein of which increased after REST knockdown. The results of this study extend previous findings, demonstrating a reciprocal control of REST expression by BMPs. Administration of exogenous BMP6 inhibits aNSC proliferation and induces the expression of the astrocytic marker glial fibrillary acidic protein, highlighting its antimitogenic and prodifferentiative effects. This study suggests that BMP6 produced in a REST-regulated manner together with other signals can contribute to regulation of NSC maintenance and fate.


Antibacterial substantivity of Carvacrol and sodium hypochlorite in infected bovine root dentin.
Various methods commonly used for cleaning and shaping root canals have not been successful in completely eradicating bacteria due to anatomic complexity and root canals irregularities. Disinfecting the canals with intracanal irrigants in addition to proper cleaning and shaping can produce a successful outcome. Antimicrobials with sustained antibacterial activity would be desirable for irrigation. The purpose of this study was to compare the antimicrobial substantivity of Carvacrol and 5.25% NaOCl in infected bovine root dentin. One hundred and twenty dentin tubes prepared from bovine incisors were infected in vitro for 14 days with Enterococcus (E) faecalis. The specimens were divided into four groups including 1) Carvacrol, 2) NaOCl, 3) infected dentin tubes (positive control); and 4) sterile dentin tubes (negative control). Dentin chips were collected at five intervals (days 0, 1, 3, 7 and 28) using round burs with sequentially increasing diameters (which includes five layers of dentin) into Brain Heart Infusion (BHI) broth. In order to compare the pre- and post-irrigation antimicrobial activity of the irrigants the colony-forming units (CFU) were counted and classified as 'CFU-before' and 'CFU-after'. After culturing, the number of CFU with the various experimental time and dentinal layers was recounted. Two-way ANOVA test was used to analyze the effects of time and materials. One-way ANOVA and supplemental Tukey HSD test were used for pair comparison. CFU was significantly reduced in NaOCl group when compared to all other experimental groups (P<0.05). The substantivity of NaOCl was significantly greater than Carvacrol. Further studies are required to investigate and approve Carvacrol as a final irrigant.


Intraperitoneal recombinant alpha 2-interferon for 'salvage' immunotherapy in persistent epithelial ovarian cancer.
Fourteen patients with epithelial ovarian cancer were treated with intraperitoneal (i.p.) administration of alpha-recombinant interferon (rIFN-alpha 2) after documentation of persistent disease at second-look laparotomy and combination chemotherapy. After therapy, 11 patients had a surgical re-evaluation which confirmed 4 complete responses (36%), 1 partial response (9%), and disease progression in 6 (55%). Five of 7 patients (71%) with minimal residual disease (MRD, i.e. less than 5 mm) had a surgically-documented response, whereas there was none in the 4 patients whose tumors were greater than or equal to 5 mm. Fever greater than or equal to 38 degrees C was seen in 58%, greater than or equal to 39.0 degrees C in 18%; nausea and vomiting in 37%, and abdominal pain in 22%. There was no consistent alteration in peripheral WBC's during treatment, while i.p. monocytes and lymphocytes showed a significant boost on day 1 after each dose of rIFN-alpha 2. Natural killer (NK) lymphocyte cytotoxicity was elevated in the i.p. cavity fluid obtained from most patients on day 1 after treatment, while blood NK values showed considerable variability. Pharmacokinetic studies showed i.p. levels of rIFN-alpha 2 were 30-1000 times blood levels. I.p. rIFN-alpha 2 may act by increasing concentrations of drug and augmenting regional host cells in patients with MRD ovarian cancer.


Dominant, toxic gain-of-function mutations in gars lead to non-cell autonomous neuropathology.
Charcot-Marie-Tooth (CMT) neuropathies are collectively the most common hereditary neurological condition and a major health burden for society. Dominant mutations in the gene GARS, encoding the ubiquitous enzyme, glycyl-tRNA synthetase (GlyRS), cause peripheral nerve degeneration and lead to CMT disease type 2D. This genetic disorder exemplifies a recurring motif in neurodegeneration, whereby mutations in essential, widely expressed genes have selective deleterious consequences for the nervous system. Here, using novel Drosophila models, we show a potential solution to this phenomenon. Ubiquitous expression of mutant GlyRS leads to motor deficits, progressive neuromuscular junction (NMJ) denervation and pre-synaptic build-up of mutant GlyRS. Intriguingly, neuronal toxicity is, at least in part, non-cell autonomous, as expression of mutant GlyRS in mesoderm or muscle alone results in similar pathology. This mutant GlyRS toxic gain-of-function, which is WHEP domain-dependent, coincides with abnormal NMJ assembly, leading to synaptic degeneration, and, ultimately, reduced viability. Our findings suggest that mutant GlyRS gains access to ectopic sub-compartments of the motor neuron, providing a possible explanation for the selective neuropathology caused by mutations in a widely expressed gene.


The two mitochondrial heat shock proteins 70, Ssc1 and Ssq1, compete for the cochaperone Mge1.
Two members of the heat shock protein 70 kDa (Hsp70) family, Ssc1 and Ssq1, perform important functions in the mitochondrial matrix. The essential Ssc1 is an abundant ATP-binding protein required for both import and folding of mitochondrial proteins. The function of Ssc1 is supported by an interaction with the preprotein translocase subunit Tim44, the cochaperone Mdj1, and the nucleotide exchange factor Mge1. In contrast, only limited information is available on Ssq1. So far, a basic characterization of Ssq1 has demonstrated its involvement in the maintenance of mitochondrial DNA, the maturation of the yeast frataxin (Yfh1) after import, and assembly of the mitochondrial Fe/S cluster. Here, we analyzed the biochemical properties and the interaction partners of Ssq1 in detail. Ssq1 showed typical chaperone properties by binding to unfolded substrate proteins in an ATP-regulated manner. Ssq1 was able to form a specific complex with the nucleotide exchange factor Mge1. In particular, complex formation in organello was enhanced significantly when Ssc1 was inactivated selectively. However, even under these conditions, no interaction of Ssq1 with the two other mitochondrial Hsp70-cochaperones, Tim44 and Mdj1, was observed. The Ssq1-Mge1 interaction showed a lower overall stability but the same characteristic nucleotide-dependence as the Ssc1-Mge1 interaction. A quantitative analysis of the interaction properties indicated a competition of Ssq1 with Ssc1 for binding to Mge1. Perturbation of Mge1 function or amounts resulted in direct effects on Ssq1 activity in intact mitochondria. We conclude that mitochondria represent the unique case where two Hsp70s compete for the interaction with one nucleotide exchange factor.


Resistance to Cyprodinil and Lack of Fludioxonil Resistance in Botrytis cinerea Isolates from Strawberry in North and South Carolina.
Chemical control of gray mold of strawberry caused by Botrytis cinerea is essential to prevent pre- and postharvest fruit decay. For more than 10 years, the anilinopyrimidine (AP) cyprodinil and the phenylpyrrole fludioxonil (Switch 62.5WG) have been available to commercial strawberry producers in the United States for gray mold control. Both active ingredients are site-specific inhibitors and, thus, prone to resistance development. In this study, 217 single-spore isolates of B. cinerea from 11 commercial strawberry fields in North and South Carolina were examined for sensitivity to both fungicides. Isolates that were sensitive (53%), moderately resistant (30%), or resistant (17%) to cyprodinil were identified based on germ tube inhibition at discriminatory doses of cyprodinil at 1 and 25 mg/liter at 10 of the 11 locations. None of the isolates was fludioxonil resistant. Phenotypes that were moderately resistant or resistant to cyprodinil were not associated with fitness penalties for mycelial growth rate, spore production, or osmotic sensitivity. Detached fruit assays demonstrated cross resistance between the two AP fungicides cyprodinil and pyrimethanil, and that isolates that were characterized in vitro as moderately resistant or resistant were equivalent in pathogenicity on fruit sprayed with pyrimethanil (currently the only AP registered in strawberry as a solo formulation). This suggests that the in vitro distinction of moderately resistant and resistant isolates is of little if any field relevance. The absence of cross-resistance with fludioxonil, iprodione, cycloheximide, and tolnaftate indicated that multidrug resistance in the form of multidrug resistance phenotypes was unlikely to be involved in conferring resistance to APs in our isolates. Implications for resistance management and disease control are discussed.


HTR1B, ADIPOR1, PPARGC1A, and CYP19A1 and obesity in a cohort of Caucasians and African Americans: an evaluation of gene-environment interactions and candidate genes.
The World Health Organization estimates that the number of obese and overweight adults has increased to 1.6 billion, with concomitant increases in comorbidity. While genetic factors for obesity have been extensively studied in Caucasians, fewer studies have investigated genetic determinants of body mass index (BMI; weight (kg)/height (m)(2)) in African Americans. A total of 38 genes and 1,086 single nucleotide polymorphisms (SNPs) in African Americans (n = 1,173) and 897 SNPs in Caucasians (n = 1,165) were examined in the Southern Community Cohort Study (2002-2009) for associations with BMI and gene × environment interactions. A statistically significant association with BMI survived correction for multiple testing at rs4140535 (β = -0.04, 95% confidence interval: -0.06, -0.02; P = 5.76 × 10(-5)) in African Americans but not in Caucasians. Gene-environment interactions were observed with cigarette smoking and a SNP in ADIPOR1 in African Americans, as well as between a different SNP in ADIPOR1 and physical activity in Caucasians. A SNP in PPARGC1A interacted with alcohol consumption in African Americans, and a different SNP in PPARGC1A was nominally associated in Caucasians. A SNP in CYP19A1 interacted with dietary energy intake in African Americans, and another SNP in CYP191A had an independent association with BMI in Caucasians.


The impact of a community-based clinical trial educational intervention among underrepresented Chinese Americans.
Clinical trials are a critical resource for the discovery of new prevention, diagnostic, and treatment methods for cancer. The most effective prevention and treatment modalities are based on previous clinical trial results. However, participation in clinical trials is underrepresented by racial/ethnic minority populations, Asian Americans in particular. Asian Americans are the least represented of any ethnic groups in clinical trials. The purpose of this study is to develop and evaluate a culturally and linguistically appropriate community-based educational intervention to increase knowledge of and intent to participate in cancer clinical trials among underrepresented Chinese Americans. Community-Based Participatory Research approach was used to guide the development, cultural tailoring, implementation, and evaluation of clinical trial intervention. First, 22 Asian community representatives were recruited as community health educators (CHE) who received 12-hour training on clinical trial education. Second, 262 members were recruited from 11 Chinese community organizations. Of those recruited, a total of 247 eligible Chinese enrolled and participated in the clinical trial education delivered by trained CHEs. Participants completed pretest before and posttest after the intervention. Fifteen of 21 measures of clinical trial knowledge showed significant changes post the intervention (P < 0.05). Education remained the sole demographic factor increasing clinical trial knowledge in multivariate analysis. Clinical trial education should emphasize both benefits to science and the larger Asian community. This community-based clinical trial intervention demonstrated promising results and has potential to enhance recruitment and participation in clinical trial research among the underrepresented Asian Americans. Improving clinical trial participation in the fast-growing Asian American population is key to dissemination of health innovations targeted to diminish health disparities.


Linking the Transcriptional Landscape of Bone Induction to Biomaterial Design Parameters.
New engineering possibilities allow biomaterials to serve as active orchestrators of the molecular and cellular events of tissue regeneration. Here, the molecular control of tissue regeneration for calcium phosphate (CaP)-based materials is established by defining the parameters critical for tissue induction and those are linked to the molecular circuitry controlling cell physiology. The material properties (microporosity, ion composition, protein adsorption) of a set of synthesized osteoinductive and noninductive CaP ceramics are parameterized and these properties are correlated to a transcriptomics profile of osteogenic cells grown on the materials in vitro. Using these data, a genetic network controlling biomaterial-induced bone formation is built. By isolating the complex material properties into single-parameter test conditions, it is verified that a subset of these genes is indeed controlled by surface topography and ions released from the ceramics, respectively. The gene network points to a decisive role for extracellular matrix deposition in osteoinduction by genes such as tenascin C and hyaluronic acid synthase 2, which are controlled by calcium and phosphate ions as well as surface topography. This work provides insight into the biomaterial composition and material engineering aspects of bone void filling and can be used as a strategy to explore the interface between biomaterials and tissue regeneration.


Ant- and Ant-Colony-Inspired ALife Visual Art.
Ant- and ant-colony-inspired ALife art is characterized by the artistic exploration of the emerging collective behavior of computational agents, developed using ants as a metaphor. We present a chronology that documents the emergence and history of such visual art, contextualize ant- and ant-colony-inspired art within generative art practices, and consider how it relates to other ALife art. We survey many of the algorithms that artists have used in this genre, address some of their aims, and explore the relationships between ant- and ant-colony-inspired art and research on ant and ant colony behavior.


Inhibition of Prostaglandin Transporter (PGT) Promotes Perfusion and Vascularization and Accelerates Wound Healing in Non-Diabetic and Diabetic Rats.
Peripheral ischemia, resulting from diminished arterial flow and defective local vascularization, is one of the main causes of impaired wound healing in diabetes. Vasodilatory prostaglandins (PGs), including PGE2 and PGI2, regulate blood flow in peripheral tissues. PGs also stimulate angiogenesis by inducing vascular endothelial growth factor. However, PG levels are reduced in diabetes mainly due to enhanced degradation. We hypothesized that inhibition of the prostaglandin transporter (PGT) (SLCO2A1), which mediates the degradation of PGs, would increase blood flow and stimulate vascularization, thereby mitigating peripheral ischemia and accelerating wound healing in diabetes. Here we report that inhibiting PGT with intravenously injected PGT inhibitor, T26A, increased blood flow in ischemic hind limbs created in non-diabetic rats and streptozotocin induced diabetic rats. Systemic, or combined with topical, T26A accelerated closure of cutaneous wounds. Immunohistochemical examination revealed that inhibition of PGT enhanced vascularization (marked by larger numbers of vessels formed by CD34+ cells), and accelerated re-epithelialization of cutaneous wounds. In cultured primary human bone marrow CD34+ cells and human epidermal keratinocytes (HEKs) either inhibiting or silencing PGT increased migration in both cell lines. Thus PGT directly regulates mobilization of endothelial progenitor cells (EPCs) and HEKs, which could contribute to PGT-mediated vascularization and re-epithelialization. At the molecular level, systemic inhibition of PGT raised circulating PGE2. Taken together, our data demonstrate that PGT modulates arterial blood flow, mobilization of EPCs and HEKs, and vascularization and epithelialization in wound healing by regulating vasodilatory and pro-angiogenic PGs.


Intraoperative transesophageal echocardiographic assessment of left ventricular Tei index in congenital heart disease.
Use of the Tei index has not been described to assess myocardial function before or after surgery in pediatric patients. This study was designed to evaluate the left ventricular (LV) function using the Tei index pre- and post-cardiopulmonary bypass in patients with lesion that result in a volume loaded right ventricle (RV). Retrospective data on 55 patients who underwent repair of a cardiac defect were analyzed. Patients with volume overload RV (n = 15) were compared to patients without volume overload but with other cardiac defects (n = 40). We reviewed pre- and post-operative LV myocardial performance index (Tei index). Tei index was obtained from transesophageal Doppler echocardiogram. Patients with right heart volume overload, the mean preoperative Tei index was 0.6, with a postoperative mean decrease of 0.207 (P = 0.014). Patients without right heart volume overload, the mean preoperative Tei was 0.48 with no significant postoperative change (P = 0.82). Pre- and post-operative transesophageal echocardiogram assessment provides an easy and quick way of evaluating LV function intra-operatively using LV Tei index. Preoperative LV Tei index was greater in the RV volume overload defects indicating diminished LV global function. This normalized in the immediate postoperative period, implying an immediate improvement in LV function. In patients without right heart volume load, consist of other cardiac defects, demonstrated no changes in the pre- and post-operative LV Tei. This implies that LV function was similar after the surgery.


Antinociceptive effects of potent, selective and brain penetrant muscarinic M4 positive allosteric modulators in rodent pain models.
Analgesic properties of orthosteric agonists of the muscarinic M4 receptor subtype have been documented in literature reports, with evidence from pharmacological and in vivo receptor knock out (KO) studies. Constitutive M4 receptor KO mice demonstrated an increased response in the formalin pain model, supporting this hypothesis. Two novel positive allosteric modulators (PAM) of the M4 receptor, Compounds 1 and 2, were characterized in rodent models of acute nociception. Results indicated decreased time spent on nociceptive behaviors in the mouse formalin model, and efficacy in the mouse tail flick assay. The analgesic-like effects of Compounds 1 and 2 were shown to be on target, as the compounds lacked any activity in constitutive M4 KO mice, while retaining activity in wild type control littermates. The analgesic-like effects of Compounds 1 and 2 were significantly diminished in KO mice that have selective deletion of the M4 receptor in neurons that co-express the dopaminergic D1 receptor subtype, suggesting a centrally-mediated effect on nociception. The opioid antagonist naloxone did not diminish the effect of Compound 1, indicating the effects of Compound 1 are not secondarily linked to opioid pathways. Compound 1 was evaluated in the rat, where it demonstrated analgesic-like effects in tail flick and a subpopulation of spinal nociceptive sensitive neurons, suggesting some involvement of spinal mechanisms of nociceptive modulation. These studies indicate that M4 PAMs may be a tractable target for pain management assuming an appropriate safety profile, and it appears likely that both spinal and supraspinal pathways may mediate the antinociceptive-like effects.


Would prostate cancer detected by screening with prostate-specific antigen develop into clinical cancer if left undiagnosed? A comparison of two population-based studies in Sweden.
To assess the risk of over-diagnosing and over-treating prostate cancer if population-based screening with serum prostate-specific antigen (PSA) is instituted. From a serum bank stored in 1980, PSA was analysed in 658 men with no previously known prostate cancer from a well-defined cohort from Göteborg, Sweden (men born in 1913); the incidence of clinical prostate cancer was registered until 1995. From the same area, and with the same selection criteria, another cohort of 710 men born in 1930-31, who in 1995 accepted an invitation for PSA screening, was also analysed. Of men born in 1913, 18 (2.7%) had died from prostate cancer and the cumulative probability of being diagnosed with clinical prostate cancer was 11.1% (5.0% in those with a PSA level of < 3 ng/mL vs 32.9% in those with a PSA level of > 3 ng/mL, P < 0.01). The mean lead-time from increased PSA (> 3 ng/mL) to clinical diagnosis was 7 years. The prostate cancer detection rate in men born in 1930-31 was 4.4% (22% among those with increased PSA levels) and 30 of 31 detected cancers were clinically localized. Screening and sextant biopsies resulted in a lower detection rate (22%) than the cumulative risk of having clinical prostate cancer (33%) in men with increased PSA levels, indicating that under-diagnosis rather than over-diagnosis is the case at least with 'one-time' screening. Even if the stage distribution in screening-detected cancers seems promising (and thus may result in reduced mortality) it is notable that screening 67-year-old men will result in treatment a mean of 7 years before clinical symptoms occur and only one in four men anticipated to develop prostate cancer will die from the disease within 15 years. Large randomized screening trials seem mandatory to further explore the benefits and hazards of PSA screening.


Detecting microdamage in bone.
Fatigue-induced microdamage in bone contributes to stress and fragility fractures and acts as a stimulus for bone remodelling. Detecting such microdamage is difficult as pre-existing microdamage sustained in vivo must be differentiated from artefactual damage incurred during specimen preparation. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye, but cutting and grinding them in an aqueous medium. Nonetheless, some artefactual cracks are partially stained and careful observation under transmitted light, or epifluorescence microscopy, is required. Fuchsin lodges in cracks, but is not site-specific. Cracks are discontinuities in the calcium-rich bone matrix and chelating agents, which bind calcium, can selectively label them. Oxytetracycline, alizarin complexone, calcein, calcein blue and xylenol orange all selectively bind microcracks and, as they fluoresce at different wavelengths and colours, can be used in sequence to label microcrack growth. New agents that only fluoresce when involved in a chelate are currently being developed--fluorescent photoinduced electron transfer (PET) sensors. Such agents enable microdamage to be quantified and crack growth to be measured and are useful histological tools in providing data for modelling the material behaviour of bone. However, a non-invasive method is needed to measure microdamage in patients. Micro-CT is being studied and initial work with iodine dyes linked to a chelating group has shown some promise. In the long term, it is hoped that repeated measurements can be made at critical sites and microdamage accumulation monitored. Quantification of microdamage, together with bone mass measurements, will help in predicting and preventing bone fracture failure in patients with osteoporosis.


Protein-engineered hydrogel encapsulation for 3-D culture of murine cochlea.
Elastin-like protein (ELP) hydrogel helps maintain the three-dimensional (3-D) cochlear structure in culture. Whole-organ culture of the cochlea is a useful model system facilitating manipulation and analysis of live sensory cells and surrounding nonsensory cells. The precisely organized 3-D cochlear structure demands a culture method that preserves this delicate architecture; however, current methods have not been optimized to serve such a purpose. A protein-engineered ELP hydrogel was used to encapsulate organ of Corti isolated from neonatal mice. Cultured cochleae were immunostained for markers of hair cells and supporting cells. Organ of Corti hair cell and supporting cell density and organ dimensions were compared between the ELP and nonencapsulated systems. These culture systems were then compared with noncultured cochlea. After 3 days in vitro, vital dye uptake and immunostaining for sensory and nonsensory cells show that encapsulated cochlea contain viable cells with an organized architecture. In comparison with nonencapsulated cultured cochlea, ELP-encapsulated cochleae exhibit higher densities of hair cells and supporting cells and taller and narrower organ of Corti dimensions that more closely resemble those of noncultured cochleae. However, we found compromised cell viability when the culture period extended beyond 3 days. We conclude that the ELP hydrogel can help preserve the 3-D architecture of neonatal cochlea in short-term culture, which may be applicable to in vitro study of the physiology and pathophysiology of the inner ear.


Testosterone production by ovarian follicles of the domestic cat (Felis catus).
In order to determine whether the ovarian follicles of the domestic cat which normally ovulates following copulation, were similar to those of the rabbit steroidogenically, the following experiments were carried out. Follicles were dissected out of ovaries from nine estrous cats throughout the year and incubated in medium alone or with luteinizing hormone (LH) or human chorionic gonadotropin (HCG). Media were removed every 15 min and stored frozen until analyzed for testosterone using established radioimmunoassay procedures. Although LH and HCG caused slight increases in testosterone production the amounts produced were much less than that produced by rabbit follicles under identical conditions. However, when follicles were incubated for a total of 8 h without removal of media, marked changes in testosterone and estradiol production occurred. These data suggest that the ovarian follicles of the estrous cat are steroidogenically active.


Acetylcholine reversal of isoproterenol-stimulated sodium currents in rabbit ventricular myocytes.
We have recently shown that beta-adrenergic agonists enhance the cardiac sodium current (INa) in rabbits through dual G-protein regulatory pathways. To determine if muscarinic cholinergic receptor stimulation can also modulate INa, we studied the effects of acetylcholine (ACh) and carbachol on INa in enzymatically dispersed rabbit ventricular myocytes. Whole-cell patch-clamp experiments done at room temperature using 20 mM [Na+]o showed that 100 nM isoproterenol increased INa and accelerated current decay as previously described. ACh (1 microM) or carbachol (1 microM) significantly reversed the stimulatory isoproterenol effects at test potentials throughout the INa activation range and at holding potentials negative to -80 mV. This effect was completely inhibited by atropine (1 microM) and was confirmed by studying single-channel INa from cell-attached patches. When INa was stimulated by forskolin (1 microM), carbachol (1 microM) significantly reversed the effect. The muscarinic-mediated inhibition of INa was inhibited by pertussis toxin (0.1 or 1.0 microgram/ml) incubation (12-15 hours), suggesting that the effect was inhibitory G-protein dependent. Further investigation of the ACh inhibitory mechanism revealed that ACh alone had no effect on INa and that when cells were dialyzed with cAMP (5 microM), ACh failed to inhibit INa. Furthermore, cGMP failed to inhibit the effect of isoproterenol on INa. These data suggest that ACh acts at or proximal to adenylate cyclase stimulation. Thus, rabbit cardiac Na+ channels are regulated by muscarinic agonists in a fashion similar to cardiac Ca2+ channels.


Acute normovolemic hemodilution in children submitted to posterior spinal fusion.
Acute Normovolemic Hemodilution (ANH) is a simple and low-cost autologous transfusion method. This study aimed at evaluating whether ANH is able to decrease homologous transfusions in children undergoing posterior spinal fusion and at verifying hemodilution-induced complications in those patients. Participated in this study 25 children, physical status ASA I and II, submitted to posterior spinal fusion and receiving ANH (group H). Hematocrit values were recorded at surgery beginning and completion, at PACU and hospital discharge. The number of children receiving homologous transfusions during and after surgery, as well as intra and postoperative complications were also recorded. Group H data were compared to other group of children submitted to the same surgical technique, however without ANH (group S). Median age and weight for both groups were, respectivelly 13 and 12 years and 41.5 and 34 kg. Immediately after anesthetic induction, 523 ml (mean) of blood were collected from group H, or the equivalent to 17.1% of volemia. Simultaneously, lactated Ringer's solution was started to decrease hematocrit to 28.8% +/- 3.72%. At surgery completion and after autologous blood infusion, hematocrit has reached mean values of 27% in group H and 30.4% in Group S (p = 0.01). At PACU and hospital discharge, differences between hematocrits were not statistically significant. Homologous blood was transfused in 28% of Group H children and 79% of Group S children (p = 0.001). There has been arterial hypotension in 28% of Group H patients and 37.5% of Group S patients (p = 0.9). Four Group S patients had severe postoperative infectious complications. Hospital stay for Groups H and S was, respectively 7.56 +/- 3.203 days for group H, and 9.75 +/- 4.245 days for group S (p = 0.009). Group H has received 3.948 +/- 1.334 ml lactated Ringer's and group S has received 2.234 +/- 953 (p = 0.0001). Normovolemic hemodilution is a safe method to decrease homologous blood needs in children submitted to posterior spinal fusion. There have been no anemia-related complications.


Parity, lactation, and breast cancer subtypes in African American women: results from the AMBER Consortium.
African American (AA) women have a disproportionately high incidence of estrogen receptor-negative (ER-) breast cancer, a subtype with a largely unexplained etiology. Because childbearing patterns also differ by race/ethnicity, with higher parity and a lower prevalence of lactation in AA women, we investigated the relation of parity and lactation to risk of specific breast cancer subtypes. Questionnaire data from two cohort and two case-control studies of breast cancer in AA women were combined and harmonized. Case patients were classified as ER+ (n = 2446), ER- (n = 1252), or triple negative (ER-, PR-, HER2-; n = 567) based on pathology data; there were 14180 control patients. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated in polytomous logistic regression analysis with adjustment for study, age, reproductive and other risk factors. ORs for parity relative to nulliparity was 0.92 (95% CI = 0.81 to 1.03) for ER+, 1.33 (95% CI = 1.11 to 1.59) for ER-, and 1.37 (95% CI = 1.06 to 1.70) for triple-negative breast cancer. Lactation was associated with a reduced risk of ER- (OR = 0.81, 95% CI = 0.69 to 0.95) but not ER+ cancer. ER- cancer risk increased with each additional birth in women who had not breastfed, with an OR of 1.68 (95% CI = 1.15 to 2.44) for 4 or more births relative to one birth with lactation. The findings suggest that parous women who have not breastfed are at increased risk of ER- and triple-negative breast cancer. Promotion of lactation may be an effective tool for reducing occurrence of the subtypes that contribute disproportionately to breast cancer mortality.


Altered uterine contractility in women with chronic endometritis.
To evaluate the alterations in endometrial waves (EW) originating from the contraction of the subendometrial myometrial layer in the periovulatory and midluteal phases in women diagnosed with chronic endometritis (CE). Case-control study. University hospital. Forty-five women referred for hysteroscopy and diagnosed with CE. Three-minute recording of transvaginal ultrasound scanning on sagittal uterine plane at periovulatory (cycle days 11-14) and midluteal phase (cycle days 19-22). Direction and frequency of EW measured by transvaginal ultrasound scan. The direction and frequency of EW were analyzed offline as accelerated (four to eight times normal speed) image sequences using video editing software, and the results were compared with 45 cycling women without CE. The EW pattern was significantly different when comparing the women with CE and controls at both the periovulatory and midluteal phases. During the periovulatory phase, we observed retrograde contractions in 26.7% versus 88%, anterograde in 24% versus 0, opposing in 22.7% versus 12%, not propagated in 13.3% versus 0, and absent in 13.3% versus 0, respectively, in the CE cases versus the control group. During the midluteal phase, we observed not propagated (41.3% vs. 61.3%), opposing (24% vs. 25.4%), absent (16.1% vs. 13.3%), anterograde (13.3% vs. 0), and retrograde (5.3% vs. 0), respectively, in the CE cases versus the control group. Women with CE show altered EW patterns in both the periovulatory and midluteal phases. Altered uterine contractility may aid in explaining the symptoms related to CE such as pain, abnormal uterine bleeding, infertility, and possibly endometriosis.


Diagnostic accuracy of SeptiFast multi-pathogen real-time PCR in the setting of suspected healthcare-associated bloodstream infection.
SeptiFast is a real-time PCR assay which targets ribosomal DNA sequences of bacteria and fungi, enabling detection and identification of the commonest pathogens in blood within a few hours, including those acquired in healthcare settings. We report here the first detailed assessment of SeptiFast that focuses on healthcare-associated bloodstream infections which develop during routine critical care. This was a prospective multicentre study designed to compare the clinical diagnostic accuracy of SeptiFast versus microbiological culture and independent clinical adjudication. This Phase III diagnostic study was performed in an adequately sized cohort of adult patients who developed new signs of suspected bloodstream infection while receiving routine critical care. Of 1,006 new episodes of suspected bloodstream infection in 853 patients, 922 (92 %) of these episodes in 795 patients met the inclusion criteria of the study. Patients had been exposed to a median of 8 days (interquartile range 4-16) of hospital care and had received high levels of organ support and recent antibiotic exposure. The SeptiFast test, when compared with bloodstream infection at the species/genus level, had a greater specificity [0.86, 95 % confidence interval (CI) 0.83-0.88] than sensitivity (0.50, 95 % CI 0.39-0.61). There was a low prevalence of blood culture-proven pathogens (9.2 %, 95 % CI 7.4-11.2 %), and the post-test probabilities of both a positive (26.3 %, 95 % CI 19.8-33.7 %) and a negative SeptiFast test (5.6 %, 95 % CI 4.1-7.4 %) indicated potential limitations of this technology in diagnosing bloodstream infection. When compared with blood culture, SeptiFast is likely to have limited utility for the diagnosis of healthcare-associated bloodstream infection in critical care patients despite its potential to deliver results more rapidly.


Development and applications of a near-infrared dye-benzylguanine conjugate to specifically label SNAP-tagged proteins.
Near-infrared (NIR) fluorescent probes are advantageous over visible ones, for they can avoid the interference from the short-wavelength background emission in biological systems. However, there are a very limited number of NIR probes that can specifically label target proteins in living cells. In this work, a series of long-wavelength dyes (N-NIR, S-NIR, and K-NIR) analogous to the novel Changsha NIR family are synthesized conveniently through a new approach that is different from the previously reported one. These three dyes have similar conjugation structures but exhibit tunable photophysical properties. N-NIR and S-NIR have large extinction coefficients over 100 000, and high fluorescence quantum yields. Although NIR absorption and emission of K-NIR are inferior to the former two, it emits in a much longer wavelength region. And all the three dyes can easily pass through the cell membranes to obtain the high-resolution NIR fluorescence images. Furthermore, N-NIR is chosen as the NIR fluorophore to develop a protein-labeling reagent PYBG-D, since it demonstrates the highest fluorescence quantum yield of up to 0.4 (in methanol). PYBG-D is efficiently synthesized through Sonogashira coupling between bromo-substituted N-NIR and alkyne-substituted benzylguanine (PYBG). The conjugate PYBG-D proves to be a specific and efficient label for O6-alkylguanine-DNA alkyltransferase (SNAP-tag) that fused to target proteins in living cells, which contributes to high resolution NIR fluorescence images under a laser confocal microscope.


The effect of all-ceramic and porcelain-fused-to-metal restorations on marginal peri-implant soft tissue color: a randomized controlled clinical trial.
The aim of this study was to test the color-change effect of all-ceramic restorations compared with porcelain-fused-to-metal (PFM) restorations on marginal peri-implant soft tissue. Thirty patients were randomly divided into 2 groups of 15 subjects each. The all-ceramic group received all-ceramic crowns on aluminum oxide-based abutments, while the PFM group received crowns on titanium or gold abutments. A reflectance spectrophotometer was used to measure the color difference (deltaE(Implant)) between the midfacial peri-implant mucosa before and after restoration insertion. The color difference (deltaE(Tooth-implant)) between the midfacial peri-implant mucosa and the gingival margin of the corresponding neighboring tooth was tested. The mucosal thickness was measured midfacially around the implant (MT(Implant)) and neighboring tooth (MT(Tooth)). deltaE(Implant) values were similar for the all-ceramic (7.4 +/- 2.7) and PFM groups (7.6 +/- 2.8). The all-ceramic group induced significantly less visible mucosal color change (3.4 +/- 1.4) compared to the PFM group (5.2 +/- 2.3). The MT(Implant) value of the all-ceramic group was 3.4 +/- 0.8 mm, while that of the PFM group was 2.9 +/- 0.9 mm, which was not significantly different. Significant differences were found when comparing MT(Implant) (3.1 +/- 0.9) and MT(Tooth) (1.2 +/- 0.3) values for test and control groups. All-ceramic restorations revealed a better color match to the neighboring teeth than PFM restorations.


The vaginal microbiome as a predictor for outcome of in vitro fertilization with or without intracytoplasmic sperm injection: a prospective study.
Is the presence or absence of certain vaginal bacteria associated with failure or success to become pregnant after an in vitro fertilization (IVF) or IVF with intracytoplasmic sperm injection (IVF-ICSI) treatment? Microbiome profiling with the use of interspace profiling (IS-pro) technique enables stratification of the chance of becoming pregnant prior to the start of an IVF or IVF-ICSI treatment. Live-birth rates for an IVF or IVF-ICSI treatment vary between 25 and 35% per cycle and it is difficult to predict who will or will not get pregnant after embryo transfer (ET). Recently, it was suggested that the composition of the vaginal microbiota prior to treatment might predict pregnancy outcome. Analysis of the vaginal microbiome prior to treatment might, therefore, offer an opportunity to improve the success rate of IVF or IVF-ICSI. In a prospective cohort study, 303 women (age, 20-42 years) undergoing IVF or IVF-ICSI treatment in the Netherlands were included between June 2015 and March 2016. Study subjects provided a vaginal sample before the start of the IVF or IVF-ICSI procedure. The vaginal microbiota composition was determined using the IS-pro technique. IS-pro is a eubacterial technique based on the detection and categorization of the length of the 16S-23S rRNA gene interspace region. Microbiome profiles were assigned to community state types based on the dominant bacterial species. The predictive accuracy of the microbiome profiles for IVF and IVF-ICSI outcome of fresh ET was evaluated by a combined prediction model based on a small number of bacterial species. From this cohort, a model was built to predict outcome of fertility treatment. This model was externally validated in a cohort of 50 women who were undergoing IVF or IVF-ICSI treatment between March 2018 and May 2018 in the Dutch division of the MVZ VivaNeo Kinderwunschzentrum Düsseldorf, Germany. In total, the vaginal microbiota of 192 women who underwent a fresh ET could be analysed. Women with a low percentage of Lactobacillus in their vaginal sample were less likely to have a successful embryo implantation. The prediction model identified a subgroup of women (17.7%, n = 34) who had a low chance to become pregnant following fresh ET. This failure was correctly predicted in 32 out of 34 women based on the vaginal microbiota composition, resulting in a predictive accuracy of 94% (sensitivity, 26%; specificity, 97%). Additionally, the degree of dominance of Lactobacillus crispatus was an important factor in predicting pregnancy. Women who had a favourable profile as well as <60% L. crispatus had a high chance of pregnancy: more than half of these women (50 out of 95) became pregnant. In the external validation cohort, none of the women who had a negative prediction (low chance of pregnancy) became pregnant. Because our study uses a well-defined study population, the results will be limited to the IVF or IVF-ICSI population. Whether these results can be extrapolated to the general population trying to achieve pregnancy without ART cannot be determined from these data. Our results indicate that vaginal microbiome profiling using the IS-pro technique enables stratification of the chance of becoming pregnant prior to the start of an IVF or IVF-ICSI treatment. Knowledge of their vaginal microbiota may enable couples to make a more balanced decision regarding timing and continuation of their IVF or IVF-ICSI treatment cycles. This study was financed by NGI Pre-Seed 2014-2016, RedMedTech Discovery Fund 2014-2017, STW Valorisation grant 1 2014-2015, STW Take-off early phase trajectory 2015-2016 and Eurostars VALBIOME grant (reference number: 8884). The employer of W.J.S.S.C. has in collaboration with ARTPred acquired a MIND subsidy to cover part of the costs of this collaboration project. The following grants are received but not used to finance this study: grants from Innovatie Prestatie Contract, MIT Haalbaarheid, other from Dutch R&D tax credit WBSO, RedMedTech Discovery Fund, (J.D.d.J.). Grants from Ferring (J.S.E.L., K.F., C.B.L. and J.M.J.S.S.), Merck Serono (K.F. and C.B.L.), Dutch Heart Foundation (J.S.E.L.), Metagenics Inc. (J.S.E.L.), GoodLife (K.F.), Guerbet (C.B.L.). R.K. is employed by ARTPred B.V. during her PhD at Erasmus Medical Centre (MC). S.A.M. has a 100% University appointment. I.S.P.H.M.S., S.A.M. and A.E.B. are co-owners of IS-Diagnostics Ltd. J.D.d.J. is co-owner of ARTPred B.V., from which he reports personal fees. P.H.M.S. reports non-financial support from ARTPred B.V. P.H.M.S., J.D.d.J. and A.E.B. have obtained patents `Microbial population analysis' (9506109) and `Microbial population analysis' (20170159108), both licenced to ARTPred B.V. J.D.d.J. and A.E.B. report patent applications `Method and kit for predicting the outcome of an assisted reproductive technology procedure' (392EPP0) and patent `Method and kit for altering the outcome of an assisted reproductive technology procedure' by ARTPred. W.J.S.S.C. received personal consultancy and educational fees from Goodlife Fertility B.V. J.S.E.L. reports personal consultancy fees from ARTPred B.V., Titus Health B.V., Danone, Euroscreen and Roche during the conduct of the study. J.S.E.L. and N.G.M.B. are co-applicants on an Erasmus MC patent (New method and kit for prediction success of in vitro fertilization) licenced to ARTPred B.V. F.J.M.B. reports personal fees from Advisory Board Ferring, Advisory Board Merck Serono, Advisory Board Gedeon Richter and personal fees from Educational activities for Ferring, outside the submitted work. K.F. reports personal fees from Ferring (commercial sponsor) and personal fees from GoodLife (commercial sponsor). C.B.L. received speakers' fee from Ferring. J.M.J.S.S. reports personal fees and other from Merck Serono and personal fees from Ferring, unrelated to the submitted paper. The other authors declare that they have no competing interests. ISRCTN83157250. Registered 17 August 2018. Retrospectively registered.


The structure and function of small nucleolar ribonucleoproteins.
Eukaryotes and archaea use two sets of specialized ribonucleoproteins (RNPs) to carry out sequence-specific methylation and pseudouridylation of RNA, the two most abundant types of modifications of cellular RNAs. In eukaryotes, these protein-RNA complexes localize to the nucleolus and are called small nucleolar RNPs (snoRNPs), while in archaea they are known as small RNPs (sRNP). The C/D class of sno(s)RNPs carries out ribose-2'-O-methylation, while the H/ACA class is responsible for pseudouridylation of their RNA targets. Here, we review the recent advances in the structure, assembly and function of the conserved C/D and H/ACA sno(s)RNPs. Structures of each of the core archaeal sRNP proteins have been determined and their assembly pathways delineated. Furthermore, the recent structure of an H/ACA complex has revealed the organization of a complete sRNP. Combined with current biochemical data, these structures offer insight into the highly homologous eukaryotic snoRNPs.


Comparison of three commonly used CT perfusion software packages in patients with acute ischemic stroke.
CT perfusion (CTP) might support decision making in patients with acute ischemic stroke by providing perfusion maps of ischemic tissue. Currently, the reliability of CTP is hampered by varying results between different post-processing software packages. The purpose of this study is to compare ischemic core volumes estimated by IntelliSpace Portal (ISP) and syngo.via with core volumes as estimated by RAPID. Thirty-five CTP datasets from patients in the MR CLEAN trial were post-processed. Core volumes were estimated with ISP using default settings and with syngo.via using three different settings: default settings (method A); additional smoothing filter (method B); and adjusted settings (method C). The results were compared with RAPID. Agreement between methods was assessed using Bland-Altman analysis and intraclass correlation coefficient (ICC). Accuracy for detecting volumes up to 25 mL, 50 mL, and 70 mL was assessed. Final infarct volumes were determined on follow-up non-contrast CT. Median core volume was 50 mL with ISP, 41 mL with syngo.via method A, 20 mL with method B, 36 mL with method C, and 11 mL with RAPID. Agreement ranged from poor (ISP: ICC 0.41; method A: ICC 0.23) to good (method B: ICC 0.83; method C: ICC 0.85). The bias (1.8 mL) and limits of agreement (-27, 31 mL) were the smallest with syngo.via with additional smoothing (method B). Agreement for detecting core volumes ≤25 mL with ISP was 54% and 57%, 85% and 74% for syngo.via methods A, B, and C, respectively. Best agreement with RAPID software is provided by syngo.via default settings with additional smoothing. Moreover, this method has the highest agreement in categorizing patients with small core volumes.


The distribution of radiolabeled corticotropin-releasing factor in pregnant rats: an investigation of placental transfer to the fetuses.
Stress during gestation can have serious consequences on the development of the fetus. Many of these effects appear to be mediated by hormones of the hypothalamic-pituitary-adrenal (HPA) axis. Corticotropin-releasing factor (CRF), released by the hypothalamus during times of stress serves to activate release of pituitary hormones and is also present in low levels in rat plasma. Moreover, the uterus contains significant quantities of CRF at implantation sites, probably from local sources. Therefore, the possibility exists that CRF may cross the placenta and activate the fetal HPA axis. However, the ability of CRF to cross the placenta has not been demonstrated. In the present study, pregnant rats were administered radiolabeled CRF intraperitoneally, and the distribution of the labeled product was determined in the fetuses and various maternal organs. High levels of activity were observed in the pregnant female's uterus, adrenals, heart and the placentae, but only background levels of activity were detected in the maternal brain. Very low levels of activity were observed in the fetuses, indicating that the transfer of CRF across the placenta is greatly restricted. These findings suggest that maternal CRF has little or no direct effect on the developing fetus during gestational stress.


Backward nonlinear surface Tamm states in left-handed metamaterials.
We addressed the existence of the nonlinear electromagnetic surface waves, the so-called Tamm states, that form at an interface separating a nonlinear uniform left-handed metamaterial and a conventional one-dimensional photonic crystal. We found two types of nonlinear surface waves, one with a hump at the interface and the other one with two humps. It was demonstrated that the nonlinear metamaterial can support the both type Tamm states with a backward energy flow and allows for a flexible control of the dispersion properties of surface states. We also, described the intensity-dependent properties of surface Tamm states for a nonlinear self-focusing medium.


Induction of carcinoembryonic-antigen-gene expression in human colorectal carcinoma by sodium butyrate.
The effect of sodium butyrate on the expression of the carcinoembryonic-antigen (CEA) gene was studied in two poorly differentiated colorectal-carcinoma cell lines (Clone-A and MIP-101) and in one well-differentiated cell line (LS-174T); A.T.C.C. no. CCL 188). Northern-blot and dot-blot analyses indicated a steady increase in CEA mRNA from day 4 to a maximal level by day 14 after these cells were exposed to 2 mM-sodium butyrate. Studies using nuclear run-off assays followed by dot-blot hybridization to a partial CEA cDNA clone demonstrated that specific increases in gene transcription rates (3-fold in MIP-101, 4-fold in LS-174T and 6-fold in Clone-A) are not sufficient to account for the observed increases in CEA mRNA abundance. Further studies showed that CEA-specific transcripts have a half-life of about 60-80 min, and treatment with sodium butyrate increased the stability of CEA-specific transcripts to about 340 min in LS-174T cells and to about 500 min in Clone-A cells. We conclude that the induction of the CEA-gene expression by sodium butyrate in colorectal-cancer cells is mediated by both transcriptional and post-transcriptional mechanisms, with CEA mRNA stability as one of the major check-points.


Phagocytic chimeric receptors require both transmembrane and cytoplasmic domains from the mannose receptor.
Phagocytosis has traditionally been viewed as a specialized function of myeloid and monocytic cells. The mannose receptor (MR) is an opsonin-independent phagocytic receptor expressed on tissue macrophages. When human MR cDNA is transfected into Cos cells, these usually non-phagocytic cells express cell surface MR and bind and ingest MR ligands such as zymosan, yeast, and Pneumocystis carinii. Expression of cDNA for Fc gamma RI (CD64), the high-affinity Fc receptor, in Cos cells confers binding but barely detectable phagocytosis of antibody-opsonized erythrocytes (EA). We report here that chimeric receptors containing the ligand-binding ectodomain of the Fc receptor and the transmembrane and cytoplasmic domains of the MR ingest bound EA very efficiently, whereas chimeras with the Fc receptor ecto- and transmembrane domains and the MR tail, or the Fc receptor ecto- and cytoplasmic domains and the MR transmembrane region, are significantly less phagocytic. All of the chimeric receptors bind ligand with equal avidity, but gain of functional phagocytosis is only conferred by the MR transmembrane and cytoplasmic domains. Endocytosis of monomeric immunoglobulin G by chimeric receptors demonstrates a similar pattern, with optimal uptake by the chimera containing both tail and transmembrane regions from the MR. The chimeric receptors with only the transmembrane or the cytoplasmic domain contributed by the MR were less efficient. Site-directed mutagenesis of the single tyrosine residue in the cytoplasmic tail (which is present in a motif homologous to an endocytosis consensus motif in the LDL receptor cytoplasmic tail [Chen, W.-J., J. L. Goldstein, and M. S. Brown. 1990. J. Biol. Chem. 265:3116]) reduces the efficiency of phagocytosis and endocytosis to a similar extent.


Reducing HIV-risk behavior among adults receiving outpatient psychiatric treatment: results from a randomized controlled trial.
This study investigated the efficacy of a 10-session, HIV-risk-reduction intervention with 221 women and 187 men receiving outpatient psychiatric care for a mental illness. Patients were randomly assigned to the HIV intervention, a structurally equivalent substance use reduction (SUR) intervention, or standard care; they were assessed pre- and postintervention and at 3- and 6-month follow-ups. Patients receiving the HIV-risk-reduction intervention reported less unprotected sex, fewer casual sex partners, fewer new sexually transmitted infections, more safer sex communications, improved HIV knowledge, more positive condom attitudes, stronger condom use intentions, and improved behavioral skills relative to patients in the SUR and control conditions. Patients receiving the SUR intervention reported fewer total and casual sex partners compared with control patients. Exploratory analyses suggested that female patients and patients diagnosed with a major depressive disorder were more likely to benefit from the HIV-risk-reduction intervention.