Comparison of extracellular protein profiles of seven serotypes of mutans streptococci grown under controlled conditions.
Extracellular proteins produced by the four human commensal species of mutans streptococci were analysed. The organisms used were Streptococcus mutans, serotypes c, e and f, Streptococcus cricetus, serotype a, Streptococcus rattus, serotype b, and Streptococcus sobrinus, serotypes d and g. They were grown in continuous culture at different generation times and pH values in media containing either glucose or fructose to determine the extent of variation in extracellular protein production that could occur for an individual strain. The results for different organisms grown under the same conditions were then compared. The total amount of protein of molecular mass greater than or equal to 60 kDa varied considerably with the growth conditions and with the strain. Generally more protein was present at a higher pH, conditions under which the organisms also form more lipoteichoic acid. With respect to individual protein components SDS-PAGE proved better than isoelectric focusing for detecting phenotypic responses by a particular strain to environmental changes and differences between the different strains. Differences in the molecular masses of protein components were particularly pronounced in the regions designated P1 (185-200 kDa), P2 (130-155 kDa) and P3 (60-95 kDa). Every strain produced at least one component in the P1 region that cross-reacted with antiserum to the purified protein from S. mutans serotype c, a protein which is indistinguishable from antigens B and I/II. Two components in the P2 region were dominant in the case of S. cricetus and S. sobrinus strains and showed glucosyltransferase (GTF) activity. GTF activity was also detected in the P3 region, particularly with S. mutans strains.


Safe practices for bed bathing in the intensive care unit: validation of a checklist.
To validate a checklist for safe bathing in critically ill patients. This is a methodological and quantitative study. Researchers developed a checklist for safe bathing in critically ill patients consisting of 41 items, which were submitted to the apparent and content validation process, evaluated by eleven judges, and interobserver reliability. For reliability analysis, the instrument was applied in 54 bed bath procedures in the ICU; Kappa and CHF tests were used. In the apparent and content validation, adjustments were made according to the judges' suggestions. Kappa values ranged from moderate to almost perfect (0.462 to 0.962), and, in some items, there was 100% agreement; the reliability of the instrument was excellent (ICC = 0.962). The instrument proved to be dependable and easy to apply. Its use will contribute to safe bed bathing and subsidize interventions aimed at increasing the quality of care.


Time series analysis demonstrates the absence of pulsatile hyphal growth.
Hyphal tip growth has been previously reported as pulsatile, defined as regularly alternating fast and slow rates of extension. The growth of pollen tubes, and hyphae of Neurospora crassa and Saprolegnia ferax were analysed using high spatial and temporal resolution. By using long (100-500 s) records of growth rate, sampled every second, it was possible to apply rigorous statistical analysis of the time series. As previously demonstrated, pollen tubes can show pulsatile growth, detectable with this system. In contrast, hyphal growth rates do not show any evidence of pulsatile growth; instead, growth rates appear to fluctuate randomly. It is concluded that pulsatile growth is not a common feature of hyphal tip growth.


The spatially varying polarization of a focused Gaussian beam in quasi-phase-matched superlattice under electro-optic effect.
We present in this paper a wave coupling theory of linear electro-optic (EO) effect for quasi-phase matched (QPM) of focused Gaussian beam in an optical superlattice (OSL). The numerical results indicate that, due to the EO effect of an appropriate applied electric field, the output beam will form spatially inhomogeneous polarization, changing continuously in transverse section of beam; the confocal parameter has a significant impact on the output polarization of Gaussian beam and determines the half-wave voltage.


Targeting Signaling Pathway Downstream of RIG-I/MAVS in the CNS Stimulates Production of Endogenous Type I IFN and Suppresses EAE.
Type I interferons (IFN), including IFNβ, play a protective role in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Type I IFNs are induced by the stimulation of innate signaling, including via cytoplasmic RIG-I-like receptors. In the present study, we investigated the potential effect of a chimeric protein containing the key domain of RIG-I signaling in the production of CNS endogenous IFNβ and asked whether this would exert a therapeutic effect against EAE. We intrathecally administered an adeno-associated virus vector (AAV) encoding a fusion protein comprising RIG-I 2CARD domains (C) and the first 200 amino acids of mitochondrial antiviral-signaling protein (MAVS) (M) (AAV-CM). In vivo imaging in IFNβ/luciferase reporter mice revealed that a single intrathecal injection of AAV-CM resulted in dose-dependent and sustained IFNβ expression within the CNS. IFNβ expression was significantly increased for 7 days. Immunofluorescent staining in IFNβ-YFP reporter mice revealed extraparenchymal CD45+ cells, choroid plexus, and astrocytes as sources of IFNβ. Moreover, intrathecal administration of AAV-CM at the onset of EAE induced the suppression of EAE, which was IFN-I-dependent. These findings suggest that accessing the signaling pathway downstream of RIG-I represents a promising therapeutic strategy for inflammatory CNS diseases, such as MS.


Congenital dyserythropoietic anemia type I (CDA I): molecular genetics, clinical appearance, and prognosis based on long-term observation.
Congenital dyserythropoietic anemia type I (CDA I) is a rare autosomal recessive disorder with ineffective erythropoiesis and iron overloading. More than 100 cases have been described, but with the exception of a report on a large Bedouin tribe, these reports include only small numbers of cases, and no data on the lifetime evolution of the disease are available. Since 1967, we have been able to follow 21 cases from 19 families for up to 37 years. Twenty-one patients with a confirmed diagnosis of CDA I exhibited chronic macrocytic anemia of variable severity, requiring regular red cell transfusions only in 2 individuals. Four developed gallstones before the age of 30 years. Fifteen of 16 cases alive at the time of analysis showed mutations of at least one allele from exons 6 to 28 within CDAN1. Iron overloading is to be expected in all patients. In 9 patients, iron depletion was started between the ages of 7 and 36 years. Splenectomy, which was performed in 7 patients, did not result in improvement of hemoglobin values. Five patients were treated with interferon alpha-2a, and all responded with a rise in hemoglobin concentration of between 25 and 35 g/L (2.5 and 3.5 g/dL) starting within 4 weeks.


Illustrations reduce contamination of midstream urine samples in the emergency department.
Urinalysis provides direction in diagnosis and treatment of patients in the emergency department (ED). Midstream urine (MSU) collection from female patients has a high contamination rate. Verbal instruction by nurses to patients reduces contamination but is inconsistent owing to lack of time and professional knowledge. This study aimed to determine if an alternative mode of instruction requiring minimal staff input may be effective. A pseudorandomised controlled trial was undertaken with 240 female patients for whom urinalysis was clinically required. No change was made to normal practice with regards to verbal instruction. Prior to collecting their sample the intervention cohort received illustrated instruction on how to collect a clean uncontaminated MSU sample. The control cohort received no illustrated instruction. Compared outcomes were rate of contamination on urinalysis, defined as 10 or more epithelial cells per high power field, and answers to a structured patient questionnaire. Contamination rate was reduced from 40% to 25% by the intervention. According to patient survey responses, verbal collection instructions were seldom given and the actions of hand washing, cleaning with a towelette, and voiding then stopping were significantly higher in the intervention group. The illustrations were well received by over 95% of patients and were considered to be clear and effective especially for patients with reading difficulties and/or from a non-English speaking background. Illustrated urine collection instructions were well accepted by female ED patients, improved the rate of proper MSU collection and reduced the rate of urinalysis contamination in the ED.


OPLS Force Field for Choline Chloride-Based Deep Eutectic Solvents.
Deep eutectic solvents (DES) are a class of solvents frequently composed of choline chloride and a neutral hydrogen bond donor (HBD) at ratios of 1:1, 1:2, or 1:3, respectively. As cost-effective and eco-friendly solvents, DESs have gained considerable popularity in multiple fields, including materials, separations, and nanotechnology. In the present work, a comprehensive set of transferable parameters have been fine-tuned to accurately reproduce bulk-phase physical properties and local intermolecular interactions for 8 different choline chloride-based DESs. This nonpolarizable force field, OPLS-DES, gave near quantitative agreement at multiple temperatures for experimental densities, viscosities, heat capacities, and surface tensions yielding overall mean absolute errors (MAEs) of ca. 1.1%, 1.6%, 5.5%, and 1.5%, respectively. Local interactions and solvent structuring between the ions and HBDs, including urea, glycerol, phenol, ethylene glycol, levulinic acid, oxalic acid, and malonic acid, were accurately reproduced when compared to radial distribution functions and coordination numbers derived from experimental liquid-phase neutron diffraction data and from first-principles molecular dynamics simulations. The reproduction of transport properties presented a considerable challenge and behaved more like a supercooled liquid near room temperature; higher-temperature simulations, e.g., 400-500 K, or an alternative polarizable force field is recommended when computing self-diffusion coefficients.


The Atg12-Atg5 conjugate has a novel E3-like activity for protein lipidation in autophagy.
Autophagy is a bulk degradation process in eukaryotic cells; autophagosomes enclose cytoplasmic components for degradation in the lysosome/vacuole. Autophagosome formation requires two ubiquitin-like conjugation systems, the Atg12 and Atg8 systems, which are tightly associated with expansion of autophagosomal membrane. Previous studies have suggested that there is a hierarchy between these systems; the Atg12 system is located upstream of the Atg8 system in the context of Atg protein organization. However, the concrete molecular relationship is unclear. Here, we show using an in vitro Atg8 conjugation system that the Atg12-Atg5 conjugate, but not unconjugated Atg12 or Atg5, strongly enhances the formation of the other conjugate, Atg8-PE. The Atg12-Atg5 conjugate promotes the transfer of Atg8 from Atg3 to the substrate, phosphatidylethanolamine (PE), by stimulating the activity of Atg3. We also show that the Atg12-Atg5 conjugate interacts with both Atg3 and PE-containing liposomes. These results indicate that the Atg12-Atg5 conjugate is a ubiquitin-protein ligase (E3)-like enzyme for Atg8-PE conjugation reaction, distinctively promoting protein-lipid conjugation.


Innervation of hyperalgesic skin in patients with complex regional pain syndrome.
To look for anatomical and histochemical signs of interaction between sensory and sympathetic nerves in the hyperalgesic skin of patients with complex regional pain syndrome. Skin samples were obtained from eight patients whose condition developed after a suspected or confirmed peripheral nerve injury, and from nine patients with features of reflex sympathetic dystrophy (RSD) following a soft-tissue injury. A skin sample was also obtained from 18 control subjects of similar age and sex distribution to patients. In patients, skin samples were taken from an area of static mechanical hyperalgesia and from an equivalent site in the contralateral limb. In controls, skin samples were obtained from the dorsum of one hand or foot. HISTOCHEMICAL MARKERS: We used neuron-specific enolase for all classes of nerve fiber; tyrosine hydroxylase for noradrenergic fibers; vasoactive intestinal peptide for sympathetic sudomotor fibers; tyrosine hydroxylase co-existing with neuropeptide Y for sympathetic vasoconstrictor fibers; and calcitonin gene-related peptide with substance P or somatostatin for peptide-containing unmyelinated sensory fibers. In patients, the distribution of markers was similar in skin taken from an area of mechanical hyperalgesia and skin taken from an equivalent site contralaterally, and was unrelated to clinical features of RSD. The distribution of markers did not differ between patients and controls. Nerve tangles immunoreactive to neuron-specific enolase, but not to other markers, were detected in samples taken from four patients and two controls. The nerve tangles were present bilaterally in two patients, and only on the affected side in two other patients. The clinical condition was more fully developed in the four patients whose skin samples contained nerve tangles than in most other patients. A major difference in distribution or change in histochemical content of cutaneous autonomic or nociceptor fibers is unlikely to underly static mechanical hyperalgesia following a soft-tissue or peripheral nerve injury. The relevance of cutaneous nerve tangles for the pathophysiology of RSD is uncertain.


High reproductive rates result in high predation risks: a mechanism promoting the coexistence of competing prey in spatially structured populations.
I tested the hypothesis that spatial structure provides a trade-off between reproduction and predation risk and thereby facilitates predator-mediated coexistence of competing prey species. I compared a cellular automata model to a mean-field model of two prey species and their common predator. In the mean-field model, the prey species with the higher reproductive rate (the superior competitor) always outcompeted the other species (the inferior competitor), both in the presence of and the absence of the predator. In the cellular automata model, both prey species, which differed only in their reproductive rates, coexisted for a long time in the presence of their common predator at intermediate levels of predation. At low predation rates, the superior competitor dominated, while high predation rates favored the inferior competitor. This discrepancy in the results of the different models was due to a trade-off that spontaneously emerged in spatially structured populations; that is, the more clustered distribution of the superior competitor made it more susceptible to predation. In addition, coexistence of competing prey species declined with increasing dispersal ranges of either prey or predator, which suggests that the trade-off that results from spatial structure becomes less important as either prey or predator disperse over a broader range.


[Psychological sequelae of abortion based on fetal indications].
Prenatal diagnosis is based on the concept of selective abortion. The termination of a pregnancy for genetic reasons means a great emotional loss followed by grief and mourning. The case reports of two women are presented, who chose to abort after the prenatal diagnosis of trisomy X. One of them showed psychological and psychosomatic symptoms as a sign of a pathological grief reaction. The depth of her grief is also shown by the published empirical investigations. Four analogous situations are discussed: abortion for psychosocial reasons, miscarriage, stillbirth and the death of premature or newborn babies. In comparison, the specific aspects of abortion for genetic reasons are: the intensive relationship to the child at the time of termination; the strains and anxiety engendered during pregnancy by medical investigations; the conflict situation without a simple solution; the personal decision and responsibility for the abortion; the compounded task of grieving and dealing with feelings of guilt. A routine follow-up for the affected women would be desirable.


Analysis and tentative structure elucidation of new anthocyanins in fruit peel of Vitis coignetiae Pulliat (meoru) using LC-MS/MS: Contribution to the overall antioxidant activity.
The skin of Vitis coignetiae Pulliat (meoru) grown wild in the Republic of Korea was analyzed for anthocyanins via HPLC coupled to ESI-MS/MS in positive ion mode. Chromatographic separation was conducted via RP HPLC using a C(18) column, with a 50-min gradient from 0 to 80% methanol in water containing 0.5% formic acid. A total of 18 anthocyanins were identified. Among them, nine compounds were newly determined by comparing the retention time (t(R)) and mass fragmentation patterns with those of the previously reported anthocyanins for other grape varieties: malvidin hexose, peonidin 3-galactoside, malvidin 3-galactoside, cyanidin, petunidin, petunidin 3-(6''-coumaroyl)-5-diglucoside, peonidin, malvidin, and malvidin 3-(6''-coumaroyl)-5-diglucoside. The antioxidant activity of the V. coignetiae Pulliat anthocyanins was determined via 2,2-diphenyl-2-picrylhydrazyl radical scavenging and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation assays in a range of concentration from 25 to 500 mg/L. The capacity increased with concentration. The IC(50) values, defined as the concentration of sample required to scavenge 50% of free radicals, were calculated as follows: 189.63+/-11.31 mg/L and 141.29+/-6.70 mg/L for 2,2-diphenyl-2-picrylhydrazyl and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation, respectively. The antioxidant activity of the V. coignetiae Pulliat anthocyanins is substantially higher than that of ascorbic acid and is similar to the effects of the extracts obtained from other grape varieties.


Antimicrobial activity of torilin isolated from Torilis japonica fruit against Bacillus subtilis.
Torilis japonica fruit has been used in therapeutic antimicrobial treatments in Korea and China since ancient times, but there is still little information on the mechanism underlying this activity. We found that the ethanol extract of T. japonica fruit showed excellent antimicrobial activity against Bacillus subtilis ATCC 6633 spores and vegetative cells. The crude ethanol extract (75%) reduced the spore concentration by 3 log cycles and the vegetative cell concentration to lower than the detection level (reduction in spore concentration by more than 6 orders of magnitude) at a concentration of 1% (w/v). The ethanol extract of T. japonica fruit was fractionated into n-hexane (H) and a water layer. The active antimicrobial compound was isolated and purified from the hexane layer, and identified as torilin (5-[1-(acetyloxy)-1-methylethyl]-3,8-dimethyl-2-oxo-1,2,4,5,6,7,8,8a-octa-hydroazulen-6-yl(2E)-2-methylbut-2-enoate; C(22)H(32)O(5)). Torilin immediately reduced vegetative cells counts by 5 to 6 orders of magnitude, and reduced spores counts by 1 order of magnitude. The integrity of structures such as the inner, middle, and outer layers of the coat and the cortex, protoplast membrane, and core are vital to spores. Torilin functions as a surfactant with hydrophilic and hydrophobic properties related to denaturalization of various proteins. The distortion of coat proteins due to direct binding polar groups of spore coats with hydrophilic groups of torilin may be responsible for the observed rapid inactivation of bacterial spores.


Biological functions of phosphatidylinositol transfer proteins.
Phosphatidylinositol/phosphatidylcholine transfer proteins (PITPs) are ubiquitous and highly conserved proteins that are believed to regulate lipid-mediated signaling events. Their ubiquity and conservation notwithstanding, PITPs remain remarkably uninvestigated. Little is known about the coupling of specific PITPs to explicit cellular functions or the mechanisms by which PITPs interface with appropriate cellular functions. The available information indicates a role for these proteins in regulating the interface between lipid metabolism and membrane trafficking in yeast, signaling in plant development, the trafficking of specialized luminal cargo in mammalian enterocytes, and neurological function in mammals. Herein, we review recent advances in PITP biology and discuss as yet unresolved issues in this field.


Bacterial teichoic acids reverse predominant IL-12 production induced by certain lactobacillus strains into predominant IL-10 production via TLR2-dependent ERK activation in macrophages.
The cytokine response of macrophages to probiotic lactobacilli varies between strains, and the balance of IL-10/IL-12 production is crucial for determination of the direction of the immune response. To clarify the mechanism whereby Lactobacillus strains differentially induce production of IL-10 and IL-12, we examined the potential relationship between cytokine production and MAPK activation. In mouse peritoneal macrophages, Lactobacillus plantarum potently induced IL-10 but weakly induced IL-12 production, whereas L. casei potently induced IL-12 but weakly induced IL-10 production. Kinetic analysis of the activation of ERK, p38, and JNK showed that L. plantarum induced a more rapid and intense activation of MAPKs, especially of ERK, than L. casei. A selective blockade of ERK activation induced by L. plantarum resulted in a decrease in IL-10 production and a simultaneous increase in IL-12 production. Interestingly, when macrophages were stimulated with a combination of L. plantarum and L. casei, IL-10 production was induced synergistically. We identified cell wall teichoic acid and lipoteichoic acid as key factors for triggering the synergistic induction of IL-10 production, although these teichoic acids alone only weakly induced IL-10 production. The effect of these teichoic acids on IL-10 production was mediated by TLR2-dependent ERK activation. Our data demonstrate that activation of the ERK pathway is critical for determination of the balance of the IL-10/IL-12 response of macrophages to lactobacilli and that predominant IL-12 production induced by certain lactobacilli such as L. casei can be converted into predominant IL-10 production when stimulated in the presence of teichoic acids.


Crumbs is required to achieve proper organ size control during Drosophila head development.
Crumbs (Crb) is a conserved apical polarity determinant required for zonula adherens specification and remodelling during Drosophila development. Interestingly, crb function in maintaining apicobasal polarity appears largely dispensable in primary epithelia such as the imaginal discs. Here, we show that crb function is not required for maintaining epithelial integrity during the morphogenesis of the Drosophila head and eye. However, although crb mutant heads are properly developed, they are also significantly larger than their wild-type counterparts. We demonstrate that in the eye, this is caused by an increase in cell proliferation that can be attributed to an increase in ligand-dependent Notch (N) signalling. Moreover, we show that in crb mutant cells, ectopic N activity correlates with an increase in N and Delta endocytosis. These data indicate a role for Crb in modulating endocytosis at the apical epithelial plasma membrane, which we demonstrate is independent of Crb function in apicobasal polarity. Overall, our work reveals a novel function for Crb in limiting ligand-dependent transactivation of the N receptor at the epithelial cell membrane.


Lacrimal sac removal made easy by methylene blue in endoscopic dacrocystorhinostomy.
Endonasal dacrocystorhinostomy (DCR) has gained interest in the last decade. Especially when a novice surgeon is performing the surgery, difficulty in identification of the lacrimal apparatus may complicate the procedure. We investigated the efficacy of methylene blue (MB) solution as a marker for lacrimal sac (LS) in endoscopic DCR. A total of 24 endoscopic DCR cases were performed (16 primary and 8 revision cases). During surgery, LS was irrigated with MB solution. Following lacrimal bone removal, medial wall of the LS is seen with a bluish hue. When the sac is dissected, the epithelium of the LS, which is heavily stained with MB contrasts with the surrounding nasal mucosa and therefore tailoring of the extent of the LS mucosa removal is facilitated. The follow-up period of the patients was 23 +/- 7.5 months. Of the 24 cases operated, only one case needed a re-operation. Use of MB solution is effective in both identifying LS and distinguishing it from surrounding nasal mucosa in DCR.


Flavonoids and platelet aggregation: A brief review.
Platelets are small anucleated fragments derived from a megakaryocyte precursor. Platelets play a key role in many physiological functions especially in hemostasis and wound healing processes in order to maintain the integrity of the circulatory system. In addition, activated platelets release cytokines and chemokines which modulate the immune response and, in some cases of hyperactivation, they could be associated to the pathogenesis of inflammatory diseases. Flavonoids are polyphenolic compounds ubiquosly found in plants known to be potent antioxidants with positive effects against diverse diseases such as cancer, neurodegenerative or cardiovascular disease. It has been reported that some flavonoids possess anti-platelet aggregation effects though different pathways, being the inhibition of the arachidonic acid-based pathway the most representative mechanism of action. In the present review, the main sources of flavonoids, as well as their bioavailability and metabolism are summarized. Moreover, the available data about the anti-aggregation effects of flavonoids and the different mechanisms of action that has been proposed until now are also discussed.


[Mechanical properties of the small intestine of rats in the normal state and after disturbed parasympathetic innervation].
In intact and vagotomized (in 14 and 30 days after the operation) rats by means of the dynamometric method values of maximal load and relative maximum elongation of the proximal and middle areas of the small intestine (SI) have been determined in vitro. Dependence of relative elongation of the SI fragments on the load applied has been investigated. The proximal part of the SI is the most firm to tearing in comparison to the middle one. Bilateral subdiaphragmatic+ truncal vagotomy results in an increased firmness to tearing and in relative maximal elongation of the SI proximal part in 14 days and in decrease of the former parameter in the same SI part in 30 days.


Delayed Oral LY333013 Rescues Mice from Highly Neurotoxic, Lethal Doses of Papuan Taipan (Oxyuranus scutellatus) Venom.
There is an unmet need for economical snakebite therapies with long shelf lives that are effective even with delays in treatment. The orally bioavailable, heat-stable, secretory phospholipase A₂ (sPLA₂) inhibitor, LY333013, demonstrates antidotal characteristics for severe snakebite envenoming in both field and hospital use. A murine model of lethal envenoming by a Papuan taipan (Oxyuranus scutellatus) demonstrates that LY333013, even with delayed oral administration, improves the chances of survival. Furthermore, LY333013 improves the performance of antivenom even after it no longer reverses neurotoxic signs. Our study is the first demonstration that neurotoxicity from presynaptic venom sPLA2S can be treated successfully, even after the window of therapeutic antivenom has closed. These results suggest that sPLA₂ inhibitors have the potential to reduce death and disability and should be considered for the initial and adjunct treatment of snakebite envenoming. The scope and capacity of the sPLA2 inhibitors ability to achieve these endpoints requires further investigation and development efforts.


Evaluation of the effect of IL-22 on human cord blood CD4+ T cells.
IL-22 is a member of IL-10 cytokine family which is believed to play an important role in inflammatory responses. IL-22 has similarities with IL-10 including conserved sequences with IL-10. IL-22 receptor is also comprised of two chains known as L-22R1 and L-10R2; supporting the speculation that the two cytokines may have similar effects. The aim of this study was to shed some light on the biological activity of IL-22 upon the cord blood CD4+CD25- T cells. In this research, cord blood T CD4+CD25- cells were cultured in presence of anti CD2/CD3/CD28 coated beads, IL-2 and IL-22 for two weeks at 37 degrees C and 5% CO2. Flow cytometry analysis showed that IL-22 has no effect upon CD25 and Foxp3 expression. Also, the results indicated that IL-22 is not involved in CD4+ T cell proliferation. Moreover, the results of suppression assay did not show any suppression effect on the cultured T cells. Thus, it seems that umbilical cord blood T cells probably do not express IL-22R1 on their surface.


Migraine-associated vertigo.
It is probably not wise to demand a temporal relationship between migraine symptoms and vertigo for the definition of migrainous vertigo. When recurrent vertigo attacks begin at an early age in a patient with normal hearing and migraine, there are few diagnoses other than migraine that need to be considered. The clinical association between migraine and vestibular symptoms, such as dizziness, motion intolerance and spontaneous attacks of vertigo, is well documented. Recently, investigators have attempted to develop diagnostic criteria for this association. We hypothesized that there are multiple migraine-associated vestibular syndromes and studied a more homogenous subset of them (benign recurrent vertigo). A structured interview was conducted over the telephone with 40 patients who presented to our neurotology clinic with benign recurrent vertigo and met the International Headache Society criteria for migraine. The structured interview was also conducted with 40 relatives of the patients who reported the same symptoms. A marked female predominance was found. Most of the patients had vertigo attacks lasting minutes or hours and most were completely free of dizziness between attacks. Imbalance and nausea typically accompanied the vertigo. However, in half of the cases, vertigo occurred without an association with headache.


Fungal infections in transplant recipients receiving alemtuzumab.
Recently, we have used an anti-T-cell agent, alemtuzumab, as induction or conversion therapy to achieve a calcineurin (CNI) and steroid-free immunosuppressive regimen. We identified recipients who developed systemic fungal infections after the initiation of alemtuzumab and looked at their outcomes. The study population consisted of all pancreas transplant recipients who received alemtuzumab. Only invasive fungal infections were included in the analysis (eg, fungemia, meningitis, or pneumonia; fungal urinary tract infections were excluded). The organism was confirmed by culture, histopathology, or latex antigen test. Between February 2003 and February 2004, a total of 121 pancreas transplant recipients received alemtuzumab-56 as part of induction, and 65 as part of conversion. Of these, 8 (6.6%) developed an invasive fungal infection; 2 (3.6%) recipients as part of induction therapy and 6 (9.2%) as part of conversion therapy. Mean recipient age was 42.1 years. The mean length of time from alemtuzumab administration (first dose) to the diagnosis of the fungal infection was 115.9 days (range 5 to 318). The organisms identified initially were: Cryptococcus, Histoplasma, Aspergillus, and Candida. Overall, 3 (38%) of the eight patients died during ongoing treatment of their fungal infection: two from sepsis, one due to myocardial infarction. The other five recipients were treated successfully and have functioning grafts. The initial therapeutic agents used included: amphotericin B/liposomal AMB (n = 6), voriconazole (n = 3), capsofungin (n = 2), and fluconazole (n = 1). The use of alemtuzumab as induction or conversion therapy in pancreas transplant recipients may predispose patients to the development of systemic fungal infections. It would be important to determine what the most appropriate prophylaxis regimen would be for these patients.


Universal selective transfer printing via micro-vacuum force.
Transfer printing of inorganic thin-film semiconductors has attracted considerable attention to realize high-performance soft electronics on unusual substrates. However, conventional transfer technologies including elastomeric transfer printing, laser-assisted transfer, and electrostatic transfer still have challenging issues such as stamp reusability, additional adhesives, and device damage. Here, a micro-vacuum assisted selective transfer is reported to assemble micro-sized inorganic semiconductors onto unconventional substrates. 20 μm-sized micro-hole arrays are formed via laser-induced etching technology on a glass substrate. The vacuum controllable module, consisting of a laser-drilled glass and hard-polydimethylsiloxane micro-channels, enables selective modulation of micro-vacuum suction force on microchip arrays. Ultrahigh adhesion switchability of 3.364 × 106, accomplished by pressure control during the micro-vacuum transfer procedure, facilitates the pick-up and release of thin-film semiconductors without additional adhesives and chip damage. Heterogeneous integration of III-V materials and silicon is demonstrated by assembling microchips with diverse shapes and sizes from different mother wafers on the same plane. Multiple selective transfers are implemented by independent pressure control of two separate vacuum channels with a high transfer yield of 98.06%. Finally, flexible micro light-emitting diodes and transistors with uniform electrical/optical properties are fabricated via micro-vacuum assisted selective transfer.


Optometry and the drug enforcement administration.
Since the passage in 1971 of the first statutory law specifically authorizing optometrist to utilize pharmaceutical agents in the practice of their profession, optometry's responsibility and relationship to the Drug Enforcement Administration (DEA) has been poorly defined. This paper describes the function of the Drug Enforcement Administration and proposes a strengthened optometric relationship with this important federal agency. Optometry's further development as a provider of primary eye and vision are services depends, in part, on this unique relationship.


Generation and characterization of induced pluripotent stem cells from breast cancer patients carrying ATM mutations.
We generated two induced pluripotent stem cell (iPSC) lines from peripheral blood mononuclear cells (PBMCs) of breast cancer patients carrying germline ATM mutations, a gene associated with a 7% prevalence in breast cancer. These iPSC lines displayed typical morphology, expressed pluripotency markers, maintained a stable karyotype, and retained the ability to differentiate into the three germ layers. These patient-specific iPSC lines hold great potential for mechanistic investigations and the development of drug screening strategies aimed at addressing ATM-related cancer.


Chromatin remodeling underlies the senescence-associated secretory phenotype of tumor stromal fibroblasts that supports cancer progression.
Age is a major risk factor for the development of cancer. Senescent fibroblasts, which accumulate with age, secrete protumorigenic factors collectively referred to as the senescence-associated secretory phenotype (SASP). Here, we examined the molecular mechanisms that control SASP activation, focusing on the known SASP factor osteopontin (OPN). We found that expression of the canonical SASP members interleukin (IL)-6 and IL-8, but not OPN, were dependent upon a persistent DNA damage response (DDR) as evidenced by ATM and NF-κB activation. Treatment with several histone deacetylase (HDAC) inhibitors robustly activated SASP in the absence of DNA breaks, suggesting that DDR-dependent SASP activation occurs in response to chromatin remodeling rather than physical breaks in DNA. In the setting of HDAC inhibition, IL-6 and IL-8 expression remained dependent upon ATM and NF-κB, while OPN expression remained independent of these factors. Further analysis revealed that HDAC1 inhibition was sufficient to induce OPN expression, which is interesting given that loss of HDAC1 expression correlates with increased OPN expression within the stromal compartment of invasive breast cancers. Importantly, fibroblasts treated with HDAC inhibitors promoted tumor growth in vivo. Our findings therefore indicate that HDAC modulation plays an important role in stromal cell activation, with important implications for the use of HDAC inhibitors in the treatment of cancer.


Enrichment of intersubtype HIV-1 recombinants in a dual infection system using HIV-1 strain-specific siRNAs.
Intersubtype HIV-1 recombinants in the form of unique or stable circulating recombinants forms (CRFs) are responsible for over 20% of infections in the worldwide epidemic. Mechanisms controlling the generation, selection, and transmission of these intersubtype HIV-1 recombinants still require further investigation. All intersubtype HIV-1 recombinants are generated and evolve from initial dual infections, but are difficult to identify in the human population. In vitro studies provide the most practical system to study mechanisms, but the recombination rates are usually very low in dual infections with primary HIV-1 isolates. This study describes the use of HIV-1 isolate-specific siRNAs to enrich intersubtype HIV-1 recombinants and inhibit the parental HIV-1 isolates from a dual infection. Following a dual infection with subtype A and D primary HIV-1 isolates and two rounds of siRNA treatment, nearly 100% of replicative virus was resistant to a siRNA specific for an upstream target sequence in the subtype A envelope (env) gene as well as a siRNA specific for a downstream target sequence in the subtype D env gene. Only 20% (10/50) of the replicating virus had nucleotide substitutions in the siRNA-target sequence whereas the remaining 78% (39/50) harbored a recombination breakpoint that removed both siRNA target sequences, and rendered the intersubtype D/A recombinant virus resistant to the dual siRNA treatment. Since siRNAs target the newly transcribed HIV-1 mRNA, the siRNAs only enrich intersubtype env recombinants and do not influence the recombination process during reverse transcription. Using this system, a strong bias is selected for recombination breakpoints in the C2 region, whereas other HIV-1 env regions, most notably the hypervariable regions, were nearly devoid of intersubtype recombination breakpoints. Sequence conservation plays an important role in selecting for recombination breakpoints, but the lack of breakpoints in many conserved env regions suggest that other mechanisms are at play. These findings show that siRNAs can be used as an efficient in vitro tool for enriching recombinants, to facilitate further study on mechanisms of intersubytpe HIV-1 recombination, and to generate replication-competent intersubtype recombinant proteins with a breadth in HIV-1 diversity for future vaccine studies.


CsrA regulates translation of the Escherichia coli carbon starvation gene, cstA, by blocking ribosome access to the cstA transcript.
CsrA is a global regulator that binds to two sites in the glgCAP leader transcript, thereby blocking ribosome access to the glgC Shine-Dalgarno sequence. The upstream CsrA binding site (GCACACGGAU) was used to search the Escherichia coli genomic sequence for other genes that might be regulated by CsrA. cstA contained an exact match that overlapped its Shine-Dalgarno sequence. cstA was previously shown to be induced by carbon starvation and to encode a peptide transporter. Expression of a cstA'-'lacZ translational fusion in wild-type and csrA mutant strains was examined. Expression levels in the csrA mutant were approximately twofold higher when cells were grown in Luria broth (LB) and 5- to 10-fold higher when LB was supplemented with glucose. It was previously shown that cstA is regulated by the cyclic AMP (cAMP)-cAMP receptor protein complex and transcribed by Esigma(70). We investigated the influence of sigma(S) on cstA expression and found that a sigma(S) deficiency resulted in a threefold increase in cstA expression in wild-type and csrA mutant strains; however, CsrA-dependent regulation was retained. The mechanism of CsrA-mediated cstA regulation was also examined in vitro. Cross-linking studies demonstrated that CsrA is a homodimer. Gel mobility shift results showed that CsrA binds specifically to cstA RNA, while coupled-transcription-translation and toeprint studies demonstrated that CsrA regulates CstA synthesis by inhibiting ribosome binding to cstA transcripts. RNA footprint and boundary analyses revealed three or four CsrA binding sites, one of which overlaps the cstA Shine-Dalgarno sequence, as predicted. These results establish that CsrA regulates translation of cstA by sterically interfering with ribosome binding.


A Comment on: Anxiety and Depression in Heart Failure: An Updated Review.
With great interest, we have read the article "Anxiety and depression in heart failure: An updated review" by Sarim Rashid et al. For its clarity and conciseness, we applaud the author for his extensive research on this delicate topic. The authors have concisely written several scenarios including the risk factors that contribute to the development and exacerbation of anxiety and depression in individuals with heart conditions, including biological, psychological, and social factors. Nonetheless, we thought it would be helpful to add a few more things to the article's conclusion to strengthen it. Exploring the pathogenesis and pathophysiology of post-sepsis related depression is of positive significance for reducing the occurrence of long-term depression and improving the prognosis of sepsis patients.


[Auditory brainstem evoked potentials in hearing loss after bacterial meningitis].
Fourteen children with acute bacterial meningitis, were examined for auditory brainstem evoked potentials (ABEP) during their initial hospital course (days 3 to 24 from admission). Different degrees of neurosensory hypoacusia were detected in 8 patients (57.14%). Children aged less than one year seemed to be more susceptible to auditory deficit. In 6 children a second ABEP was done after a time interval which ranged from 20 to 570 days after the first one and all them showed residual hypoacusia. The short term evolution suggests that the acute process can be prolonged for more than 1 month after hospital admittance, and the altered auditory function tends to persist over the mid term. It is concluded that despite this research's methodological limitations. ABEP seem useful for early detection of postmeningitical neurosensory hypoacusia and follow-up of these patients.


Phonological investigation of verbal autistic and mentally retarded subjects.
A review of the pertinent literature indicates that autistic children are likely to show normal but delayed development of speech sounds. In contrast, atypical phonological development is suggested by experiments demonstrating that autistic subjects are deficient in their ability to extract the components of structured auditory input. A systematic investigation of the speech sound systems of verbal autistic and mentally retarded children reveals a delay in phoneme acquisition, as well as a relative uniformity of error types in both groups. The autistic subjects, however, differ significantly from the mentally retarded in the phonemic substitutions which they make. Autistic subjects are also characterized by a high correlation between frequency of phonological errors and level of overall language development. The findings are interpreted as supporting the hypothesis that the autistic group shows a more global delay in language development.


Laboratory automation: trajectory, technology, and tactics.
Laboratory automation is in its infancy, following a path parallel to the development of laboratory information systems in the late 1970s and early 1980s. Changes on the horizon in healthcare and clinical laboratory service that affect the delivery of laboratory results include the increasing age of the population in North America, the implementation of the Balanced Budget Act (1997), and the creation of disease management companies. Major technology drivers include outcomes optimization and phenotypically targeted drugs. Constant cost pressures in the clinical laboratory have forced diagnostic manufacturers into less than optimal profitability states. Laboratory automation can be a tool for the improvement of laboratory services and may decrease costs. The key to improvement of laboratory services is implementation of the correct automation technology. The design of this technology should be driven by required functionality. Automation design issues should be centered on the understanding of the laboratory and its relationship to healthcare delivery and the business and operational processes in the clinical laboratory. Automation design philosophy has evolved from a hardware-based approach to a software-based approach. Process control software to support repeat testing, reflex testing, and transportation management, and overall computer-integrated manufacturing approaches to laboratory automation implementation are rapidly expanding areas. It is clear that hardware and software are functionally interdependent and that the interface between the laboratory automation system and the laboratory information system is a key component. The cost-effectiveness of automation solutions suggested by vendors, however, has been difficult to evaluate because the number of automation installations are few and the precision with which operational data have been collected to determine payback is suboptimal. The trend in automation has moved from total laboratory automation to a modular approach, from a hardware-driven system to process control, from a one-of-a-kind novelty toward a standardized product, and from an in vitro diagnostics novelty to a marketing tool. Multiple vendors are present in the marketplace, many of whom are in vitro diagnostics manufacturers providing an automation solution coupled with their instruments, whereas others are focused automation companies. Automation technology continues to advance, acceptance continues to climb, and payback and cost justification methods are developing.


Extracellular pH modulates GABAergic neurotransmission in rat hypothalamus.
Changes in extracellular pH have a modulatory effect on GABAA receptor function. It has been reported that pH sensitivity of the GABA receptor is dependent on subunit composition and GABA concentration. Most of previous investigations focused on GABA-evoked currents, which only reflect the postsynaptic receptors. The physiological relevance of pH modulation of GABAergic neurotransmission is not fully elucidated. In the present studies, we examined the influence of extracellular pH on the GABAA receptor-mediated inhibitory neurotransmission in rat hypothalamic neurons. The inhibitory postsynaptic currents (IPSCs), tonic currents, and the GABA-evoked currents were recorded with whole-cell patch techniques on the hypothalamic slices from Sprague-Dawley rats at 15-26 postnatal days. The amplitude and frequency of spontaneous GABA IPSCs were significantly increased while the external pH was changed from 7.3 to 8.4. In the acidic pH (6.4), the spontaneous GABA IPSCs were reduced in amplitude and frequency. The pH induced changes in miniature GABA IPSCs (mIPSCs) similar to that in spontaneous IPSCs. The pH effect on the postsynaptic GABA receptors was assessed with exogenously applied varying concentrations of GABA. The tonic currents and the currents evoked by sub-saturating concentration of GABA ([GABA]) (10 μM) were inhibited by acidic pH and potentiated by alkaline pH. In contrast, the currents evoked by saturating [GABA] (1mM) were not affected by pH changes. We also investigated the influence of pH buffers and buffering capacity on pH sensitivity of GABAA receptors on human recombinant α1β2γ2 GABAA receptors stably expressed in HEK 293 cells. The pH influence on GABAA receptors was similar in HEPES- and MES-buffered media, and not dependent on protonated buffers, suggesting that the observed pH effect on GABA response is a specific consequence of changes in extracellular protons. Our data suggest that the hydrogen ions suppress the GABAergic neurotransmission, which is mediated by both presynaptic and postsynaptic mechanisms.


Impact of disability on quality of life of rural disabled people in Bangladesh.
This study examined the impact of disability on the quality of life of disabled people in rural Bangladesh. A primary healthcare specialist conducted a door-to-door survey in two villages in Bangladesh to collect socioeconomic and demographic information on the villagers and for identification of disabled people. Information on disability and how it affected their life was also obtained either from the disabled people or from their caregivers by interviewing them. The study revealed that disability had a devastating effect on the quality of life of the disabled people with a particularly negative effect on their marriage, educational attainment, employment, and emotional state. Disability also jeopardized their personal, family and social life. More than half of the disabled people were looked at negatively by society. Disabled women and girl children suffered more from negative attitudes than their male counterparts, resulting in critical adverse effects on their psychological and social health. A combination of educational, economic and intensive rehabilitative measures should be implemented urgently to make them self-reliant. Collaborative communication between professionals and parents, behavioural counselling, formation of a self-help group, and comprehensive support to families will reduce their suffering.


A substance use decision aid for medically at-risk adolescents: results of a randomized controlled trial for cancer-surviving adolescents.
Adolescent survivors of childhood cancer engage in risky behaviors. This study tested a decision aid for cancer-surviving adolescents aimed at difficult decisions related to engaging in substance use behaviors. This randomized controlled trial recruited 243 teen survivors at 3 cancer centers. The cognitive-behavioral skills program focused on decision making and substance use within the context of past treatment. Effects at 6 and 12 months were examined for decision making, risk motivation, and substance use behaviors using linear regression models. The majority of the teen cancer survivors (90%) rated the program as positive. There was an intermediate effect at 6 months for change in risk motivation for low riskers, but this effect was not sustained at 12 months. For quality decision making, there was no significant effect between treatment groups for either time point. The overall program effects were modest. Once teen survivors are in the program and learn what quality decision making is, their written reports indicated adjustment in their perception of their decision-making ability; thus, a more diagnostic baseline decision-making measure and a more intensive intervention are needed in the last 6 months. With 2 of 3 teen participants dealing with cognitive difficulties, the data suggest that this type of intervention will continue to be challenging, especially when 90% of their household members and 56% of their close friends model substance use. This effectiveness trial using late-effects clinics provides recommendations for further program development for medically at-risk adolescents, particularly ones with cognitive difficulties.


Internal carbonic anhydrase activity in the tissue of scleractinian corals is sufficient to support proposed roles in photosynthesis and calcification.
Reef-building corals import inorganic carbon (Ci) to build their calcium carbonate skeletons and to support photosynthesis by the symbiotic algae that reside in their tissue. The internal pathways that deliver Ci for both photosynthesis and calcification are known to involve the enzyme carbonic anhydrase (CA), which interconverts CO2 and HCO3 (-). We have developed a method for absolute quantification of internal CA (iCA) activity in coral tissue based on the rate of (18)O-removal from labeled Ci. The method was applied to three Caribbean corals (Orbicella faveolata, Porites astreoides and Siderastrea radians) and showed that these species have similar iCA activities per unit surface area, but that S. radians has ∼10-fold higher iCA activity per unit tissue volume. A model of coral Ci processing shows that the measured iCA activity is sufficient to support the proposed roles for iCA in Ci transport for photosynthesis and calcification. This is the case even when iCA activity is homogeneously distributed throughout the coral, but the model indicates that it would be advantageous to concentrate iCA in the spaces where calcification (the calcifying fluid) and photosynthesis (the oral endoderm) take place. We argue that because the rates of photosynthesis and calcification per unit surface area are similar among the corals studied here, the areal iCA activity used to deliver Ci for these reactions should also be similar. The elevated iCA activity per unit volume of S. radians compared with that of the other species is probably due to the thinner effective tissue thickness in this species.


Difficulty of acute hepatitis C diagnosis in a hospitalised patient.
The hepatitis C virus (HCV) causes acute hepatitis C and is commonly detected via HCV antibody testing. However, delayed seroconversion of HCV antibodies and non-specific symptoms may hinder the diagnosis of this disease. A 71-year-old woman developed acute hepatitis while hospitalised for back pain. An HCV antibody test was negative, and she had no risk factors for hepatitis C. She was referred to our hospital for further evaluation. The HCV antibody test was repeated 16 days after the initial test; owing to a positive result, she was diagnosed with acute hepatitis C. Several months thereafter, the HCV spontaneously cleared. When diagnosing an HCV infection, the time at which the testing is performed needs to coincide with the time at which HCV antibody seroconversion occurs. Timely diagnosis of an HCV infection allows appropriate treatment during the acute phase which may prevent disease progression to the chronic phase.


The big picture of recovery: a systematic review on the evidence of photography-based methods in researching recovery from mental distress.
In the context of a growing body of literature on incorporating visual media in researching well-being and mental health, this systematic review examined the evidence of using photography-based research methods in exploring recovery from mental distress, their outcomes, but also limitations and challenges encountered by researchers. Six cross-disciplinary electronic databases (CINAHL, MEDLINE, Web of Science, Scopus, PsycINFO, Arts & Humanities) were systematically searched resulting in a total of 15 qualitative and mixed-methods studies included in a thematic synthesis. Photo-elicitation and photovoice were identified as the main photographic methods employed in recovery research along with less common, but nonetheless creative, techniques. Four key themes were identified through thematic analysis in photography-based recovery studies: enhanced understanding, collaboration and empowerment, situatedness, and storytelling. The results of this review revealed photography as a valuable methodological tool with potential to contribute to conceptualising recovery from the stance of research participants, but also facilitate and support their recovery processes. Recovery research can benefit from the use of photographic methods that are widely accessible, versatile, and interactive. They may offer mental health researchers alternative ways to explore individuals' perspective on recovery in ways that are creative, empowering, and supportive of their recovery.


Functional changes of the liver in the absence of growth hormone (GH) action - Proteomic and metabolomic insights from a GH receptor deficient pig model.
The liver is a central target organ of growth hormone (GH), which stimulates the synthesis of insulin-like growth factor 1 (IGF1) and affects multiple biochemical pathways. A systematic multi-omics analysis of GH effects in the liver has not been performed. GH receptor (GHR) deficiency is a unique model for studying the consequences of lacking GH action. In this study, we used molecular profiling techniques to capture a broad spectrum of these effects in the liver of a clinically relevant large animal model for Laron syndrome. We performed holistic proteome and targeted metabolome analyses of liver samples from 6-month-old GHR-deficient (GHR-KO) pigs and GHR-expressing controls (four males, four females per group). GHR deficiency resulted in an increased abundance of enzymes involved in amino acid degradation, in the urea cycle, and in the tricarboxylic acid cycle. A decreased ratio of long-chain acylcarnitines to free carnitine suggested reduced activity of carnitine palmitoyltransferase 1A and thus reduced mitochondrial import of fatty acids for beta-oxidation. Increased levels of short-chain acylcarnitines in the liver and in the circulation of GHR-KO pigs may result from impaired beta-oxidation of short-chain fatty acids or from increased degradation of specific amino acids. The concentration of mono-unsaturated glycerophosphocholines was significantly increased in the liver of GHR-KO pigs without morphological signs of steatosis, although the abundances of several proteins functionally linked to non-alcoholic fatty liver disease (fetuin B, retinol binding protein 4, several mitochondrial proteins) were increased. Moreover, GHR-deficient liver samples revealed distinct changes in the methionine and glutathione metabolic pathways, in particular, a significantly increased level of glycine N-methyltransferase and increased levels of total and free glutathione. Several proteins revealed a sex-related abundance difference in the control group but not in the GHR-KO group. Our integrated proteomics/targeted metabolomics study of GHR-deficient and control liver samples from a clinically relevant large animal model identified a spectrum of biological pathways that are significantly altered in the absence of GH action. Moreover, new insights into the role of GH in the sex-related specification of liver functions were provided.


The cardiovascular and autonomic effects of repeated administration of delta-9-tetrahydrocannabinol to rhesus monkeys.
The cardiovascular effects of repeated dosing with delta-9-tetrahydrocannabinol (THC) were examined in chronically catheterized, conscious rhesus monkeys. THC (0.5 mg/kg) was administered i.v. every 6 hr for 3 weeks. Following the first THC bolus, a tachycardia (peak change, 34 beats/min) and hypotension (peak change, -21 mm Hg systolic blood pressure) sometimes preceded by a transient blood pressure increase were observed. Tolerance developed to the tachycardia and systolic blood pressure increase but not to the blood pressure decrease. Hourly measurement of systolic blood pressure and heart rate during the intervals between injections revealed heart rate decreases to which tolerance developed and blood pressure decreases which were sustained throughout the course of TCH dosing. Urinary catecholamine excretion increased during the 1st week of dosing and subsequently returned toward base-line levels. Control of basal heart rate and reflex heart rate changes was examined using autonomic agonists and antagonists. Heart rate decreases after beta adrenergic blockage (sotalol HCl) were attenuated, whereas heart rate increases after cholinergic blockade (atropine methylnitrate increases after cholinergic blockade (atropine methylnitrate) were augmented. Baroreceptor gain, measured as the ratio of heart rate change to systolic blood pressure change in response to phenylephrine, increased. The response of basal heart rate to antagonists and the augmented reflex bradycardia suggest that THC alters autonomic control of the cardiovascular system resulting in parasympathetic dominance. Divergent patterns of development of tolerance to heart rate and blood pressure effects suggest that THC affects heart rate and blood pressure through different mechanisms.


Intercellular communication in the mammalian ovary: oocytes carry the conversation.
The production of functional female gametes is essential for the propagation of all vertebrate species. The growth of oocytes within ovarian follicles and their development to mature eggs have fascinated biologists for centuries, and scientists have long realized the importance of the ovarian follicle's somatic cells in nurturing oogenesis and delivering the oocyte to the oviduct by ovulation. Recent studies have revealed key roles of the oocyte in folliculogenesis and established that bidirectional communication between the oocyte and companion somatic cells is essential for development of an egg competent to undergo fertilization and embryogenesis. The challenge for the future is to identify the factors that participate in this communication and their mechanisms of action.


Ultrahigh numerical aperture meta-fibre for flexible optical trapping.
Strong focusing on diffraction-limited spots is essential for many photonic applications and is particularly relevant for optical trapping; however, all currently used approaches fail to simultaneously provide flexible transportation of light, straightforward implementation, compatibility with waveguide circuitry, and strong focusing. Here, we demonstrate the design and 3D nanoprinting of an ultrahigh numerical aperture meta-fibre for highly flexible optical trapping. Taking into account the peculiarities of the fibre environment, we implemented an ultrathin meta-lens on the facet of a modified single-mode optical fibre via direct laser writing, leading to a diffraction-limited focal spot with a record-high numerical aperture of up to NA ≈ 0.9. The unique capabilities of this flexible, cost-effective, bio- and fibre-circuitry-compatible meta-fibre device were demonstrated by optically trapping microbeads and bacteria for the first time with only one single-mode fibre in combination with diffractive optics. Our study highlights the relevance of the unexplored but exciting field of meta-fibre optics to a multitude of fields, such as bioanalytics, quantum technology and life sciences.


The human thymic microenvironment: thymic epithelium contains specific keratins associated with early and late stages of epidermal keratinocyte maturation.
Normal T-cell development is dependent on interactions with the thymic microenvironment; thymic epithelial cells are thought to play a key role in the induction of thymocyte maturation, both through direct contact and, indirectly, via thymic hormone secretion. It has been postulated that thymic epithelial cells progress through an antigenically defined pathway of differentiation similar to that of epidermal keratinocytes. As keratins vary according to epithelial cell type and the stage of epithelial cell maturation, we used a panel of monoclonal antibodies against keratins to study specific types of keratin intermediate filaments within human thymic epithelium. The demonstration in human thymus of keratins previously shown to be associated with distinct stages of epidermal keratinocytic maturation would support the hypothesis that thymic epithelial cells undergo sequential stages of differentiation. Two-dimensional immunoblot analysis of cytoskeletal extracts from human thymus revealed that thymic epithelium contains the following keratins: 1-2, 5, 6, 7, 8, 10, 13, 14, 15, 16, and 17 (molecular masses, 65-67, 58, 56, 54, 52, 56.5, 51, 50, 50', 48, and 46 kilodaltons, respectively). Thus, in thymic epithelium, we found keratins previously observed in epidermal basal cells (5, 14, 15), as well as keratins specific for terminally differentiated keratinocytes in supra-basal epidermis (1-2, 10). Indirect immunofluorescence (IF) performed on fetal and postnatal human thymus demonstrated that keratin epitopes recognized by antibodies AE-3, 35 beta H11, and RTE-23 are present on epithelial cells of the subcapsular cortex, the cortex, the medulla, and Hassall's bodies. In contrast, antibodies AE-1 and RTE-22 reacted primarily with neuroendocrine thymic epithelium (subcapsular cortex, medulla, Hassall's bodies). The epithelial reactivity of antibody AE-2 was limited to epithelial cells in Hassall's bodies and did not appear until 16 weeks of fetal gestation i.e., when Hassall's bodies first formed. Two-dimensional gel analysis of thymic keratins demonstrated that antibody AE-2 identified only the keratins with molecular masses of 56.6 and 65-67 kilodaltons (10 and 1-2 respectively) in thymus. These data, together with the selective reactivity of AE-2 with Hassall's bodies in fluorescence assays, demonstrate the localization in Hassall's bodies of the high-molecular-weight keratins associated with the late stages of epidermal cell maturation. In summary, we demonstrated that human thymic epithelium contains specific keratins found in multiple epithelial types as well as keratins associated with both early and late stages of epidermal cell differentiation.(ABSTRACT TRUNCATED AT 400 WORDS)


Indirect Transmission of Influenza A Virus between Pig Populations under Two Different Biosecurity Settings.
Respiratory disease due to influenza virus is common in both human and swine populations around the world with multiple transmission routes capable of transmitting influenza virus, including indirect routes. The objective of this study was to evaluate the role of fomites in influenza A virus (IAV) transmission between pig populations separated by two different biosecurity settings. Thirty-five pigs were divided into four experimental groups: 10 pigs (1 replicate) were assigned to the infected group (I), 10 pigs (2 replicates of 5 pigs) were assigned to the low biosecurity sentinel group (LB), 10 pigs (2 replicates of 5 pigs) were assigned to the medium biosecurity sentinel group (MB), and 5 pigs (1 replicate) were assigned to the negative control group (NC). Eight of 10 pigs in the infected group were inoculated with IAV and 36 hours following inoculation, personnel movement events took place in order to move potentially infectious clothing and personal protective equipment (PPE) to sentinel pig rooms. Following contact with the infected group, personnel moved to the MB group after designated hygiene measures while personnel moved directly to the LB group. Nasal swabs and blood samples were collected from pigs to assess IAV infection status and fomites were sampled and tested via RRT-PCR. All experimentally inoculated pigs were infected with IAV and 11 of the 144 fomite samples collected following contact with infected pigs were low level positive for IAV genome. One replicate of each sentinel groups LB and MB became infected with IAV and all five pigs were infected over time. This study provides evidence that fomites can serve as an IAV transmission route from infected to sentinel pigs and highlights the need to focus on indirect routes as well as direct routes of transmission for IAV.


Multicenter, double-blind, randomized, intraindividual crossover comparison of gadobenate dimeglumine and gadopentetate dimeglumine for MR angiography of peripheral arteries.
To prospectively compare the image quality and diagnostic performance achieved with doses of gadobenate dimeglumine and gadopentetate dimeglumine of 0.1 mmol per kilogram of body weight in patients undergoing contrast material-enhanced magnetic resonance (MR) angiography of the pelvis, thigh, and lower-leg (excluding foot) for suspected or known peripheral arterial occlusive disease. Institutional review board approval was granted from each center and informed written consent was obtained from all patients. Between November 2006 and January 2008, 96 patients (62 men, 34 women; mean age, 63.7 years +/- 10.4 [standard deviation]; range, 39-86 years) underwent two identical examinations at 1.5 T by using three-dimensional spoiled gradient-echo sequences and randomized 0.1-mmol/kg doses of each agent. Images were evaluated on-site for technical adequacy and quality of vessel visualization and offsite by three independent blinded readers for anatomic delineation and detection/exclusion of pathologic features. Comparative diagnostic performance was determined in 31 patients who underwent digital subtraction angiography. Data were analyzed by using the Wilcoxon signed-rank, McNemar, and Wald tests. Interreader agreement was determined by using generalized kappa statistics. Differences in quantitative contrast enhancement were assessed and a safety evaluation was performed. Ninety-two patients received both agents. Significantly better performance (P < .0001; all evaluations) with gadobenate dimeglumine was noted on-site for technical adequacy and vessel visualization quality and offsite for anatomic delineation and detection/exclusion of pathologic features. Contrast enhancement (P < or = .0001) and detection of clinically relevant disease (P < or = .0028) were significantly improved with gadobenate dimeglumine. Interreader agreement for stenosis detection and grading was good to excellent (kappa = 0.749 and 0.805, respectively). Mild adverse events were reported for four (six events) and five (eight events) patients after gadobenate dimeglumine and gadopentetate dimeglumine, respectively. Higher-quality vessel visualization, greater contrast enhancement, fewer technical failures, and improved diagnostic performance are obtained with gadobenate dimeglumine, relative to gadopentetate dimeglumine, when compared intraindividually at 0.1-mmol/kg doses in patients undergoing contrast-enhanced MR angiography for suspected peripheral arterial occlusive disease.


Detecting host responses to microbial stimulation using primary epithelial organoids.
The intestinal epithelium is constantly exposed to microbes residing in the lumen. Traditionally, the response to microbial interactions has been studied in cell lines derived from cancerous tissues, e.g. Caco-2. It is, however, unclear how the responses in these cancer cell lines reflect the responses of a normal epithelium and whether there might be microbial strain-specific effects. To address these questions, we derived organoids from the small intestine from a cohort of healthy individuals. Culturing intestinal epithelium on a flat laminin matrix induced their differentiation, facilitating analysis of microbial responses via the apical membrane normally exposed to the luminal content. Here, it was evident that the healthy epithelium across multiple individuals (n = 9) demonstrates robust acute both common and strain-specific responses to a range of probiotic bacterial strains (BB-12Ⓡ, LGGⓇ, DSM33361, and Bif195). Importantly, parallel experiments using the Caco-2 cell line provide no acute response. Collectively, we demonstrate that primary epithelial cells maintained as organoids represent a valuable resource for assessing interactions between the epithelium and luminal microbes across individuals, and that these models are likely to contribute to a better understanding of host microbe interactions.


[Clinical interventions on the major vascular risk factors (RIVANA study)].
To describe the diagnostic frequency of classical vascular risk factors, dietary and therapeutic interventions and their control. Within the project Vascular Risk in Navarre (RIVANA), a transversal study was made of a random sample of 4,354 people between the ages of 35 and 84; antecedents were gathered by interview and clinical and analytical exploration was carried out. The subjects were classified for each risk factor into: diagnosed by information--when health personnel had informed them that they presented a risk factor; by intervention--when they take medication for it; and by detection in asymptomatic people. The variables of intervention were identified by dichotomic trees expressed by relative frequencies. Forty-five point three percent of the subjects are hypertense: 27% previously diagnosed and 18.3% at the time of the exploration. Thirty-nine percent are hypercholesterolemic: 33.4% previously diagnosed and 5.6% at the time of the exploration. Twenty-one point two percent presented hyperglycemia: 9.7% previously diagnosed and 11.6% at the time of the exploration. Sixty-five point six percent were diagnosed as overweight or obese: 25.9% previously and 39.6% at the time of the exploration. They received dietary counselling and pharmacological treatment respectively: 79.8% and 70.3% of the hypertense; 75.4% and 35% of the hypercholesterolemic; 83.1% and 47.4% of those who presented hyperglycemia and 68.5% and 4% of those who showed overweight or obesity. The frequency of cardiovascular risk factors is high. There is a margin for significant improvement in the detection, intervention and control of the risk factors.


Dietary factors and thyroid cancer risk: a meta-analysis of observational studies.
To better understand the relationship between dietary factors and thyroid cancer risk, we summarized the published evidence on relationship between dietary factors and thyroid cancer incidence. Searching several databases for relevant studies published by March 2014 included a total of 19 studies. We calculated summary odds ratios (ORs) for each risk factor. Based on the highest level of total consumption vs. the lowest level, the summary OR [95% (confidence interval) CI] of thyroid cancer was 0.79 (0.66, 0.94) for fish; 0.95 (0.74, 1.23) for salt water fish; 0.86 (0.63, 1.16) for fresh water fish; 0.76 (0.58, 1.00) for vegetables; 0.88 (0.72, 1.08) for shellfish; 0.93 (0.66, 1.29) for cruciferous vegetables; 0.97 (0.78, 1.21) for fruits; 0.96 (0.70, 1.34) for meat; and 1.11 (0.86, 1.42) for grains. Subgroup analysis showed that fish (OR 0.74, 95%CI: 0.59, 0.92) and shellfish (OR 0.46, 95%CI: 0.27, 0.75) consumption have a protective effect in iodine deficiency areas, whereas the ORs were not statistically significant in iodine-rich areas. Our findings indicated that fish and shellfish consumption may decrease the risk of thyroid cancer in iodine deficiency areas, although no such effect was observed in iodine-rich areas.


Peripheral blood in Sjögren's syndrome does not contain increased levels of T lymphocytes reactive with the recombinant Ro/SS-A 52 kD and La/SS-B 48 kD autoantigens.
Patients with Sjögren's syndrome (SS) frequently have anti-Ro/SS-A and anti-La/SS-B autoantibodies. The aim of this study was to investigate if these patients have peripheral blood lymphocytes (PBL) secreting IFN-gamma after short-term cultivation in the presence of Ro/SS-A and La/SS-B antigens. The frequency of PBL secreting IFN-gamma was examined in 12 SS patients and 11 healthy controls. The enzyme-linked immunospot (ELISPOT) assay was performed after 48 hours cultivation of PBL in the presence of recombinant Ro 52 kD protein or recombinant La 48 kD protein. The number of unstimulated IFN-gamma secreting cells in the SS patient group was not significantly different from that of the control group. Moreover, no increase in the number of IFN-gamma secreting cells after Ro/SS-A and La/SS-B stimulation was detected in the two groups. Thus, T cells reactive with the recombinant Ro 52 kD and La 48 kD proteins do not occur with any increased frequency in peripheral blood of SS patients.


Assessing contaminant sensitivity of endangered and threatened aquatic species: part I. Acute toxicity of five chemicals.
Assessment of contaminant impacts to federally identified endangered, threatened and candidate, and state-identified endangered species (collectively referred to as "listed" species) requires understanding of a species' sensitivities to particular chemicals. The most direct approach would be to determine the sensitivity of a listed species to a particular contaminant or perturbation. An indirect approach for aquatic species would be application of toxicity data obtained from standard test procedures and species commonly used in laboratory toxicity tests. Common test species (fathead minnow, Pimephales promelas; sheepshead minnow, Cyprinodon variegatus; and rainbow trout, Oncorhynchus mykiss) and 17 listed or closely related species were tested in acute 96-hour water exposures with five chemicals (carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin) representing a broad range of toxic modes of action. No single species was the most sensitive to all chemicals. For the three standard test species evaluated, the rainbow trout was more sensitive than either the fathead minnow or sheepshead minnow and was equal to or more sensitive than listed and related species 81% of the time. To estimate an LC50 for a listed species, a factor of 0.63 can be applied to the geometric mean LC50 of rainbow trout toxicity data, and more conservative factors can be determined using variance estimates (0.46 based on 1 SD of the mean and 0.33 based on 2 SD of the mean). Additionally, a low- or no-acute effect concentration can be estimated by multiplying the respective LC50 by a factor of approximately 0.56, which supports the United States Environmental Protection Agency approach of multiplying the final acute value by 0.5 (division by 2). When captive or locally abundant populations of listed fish are available, consideration should be given to direct testing. When direct toxicity testing cannot be performed, approaches for developing protective measures using common test species toxicity data are available.


Global harmonization of herbal health claims.
Over the past decade, herbal medicine has become a topic of increasing global importance, with both medical and economic implications. In developing countries, as much as 80% of the indigenous populations depends on traditional systems of medicine and medicinal plants as their primary source of healthcare. Within the European Community, herbal medicines represent an important share of the pharmaceutical market, with annual sales in the range of US$7 billion. In the United States, the sale of herbal products has skyrocketed from $200 million in 1988 to >$3.3 billion in 1997. Such widespread use of botanicals throughout the world has raised serious questions concerning the quality, safety and efficacy of these products. Thus, accurate scientific assessment of herbal medicine is a prerequisite for global harmonization of herbal health claims. In 1995, as part of its overall global strategy of "Health for All" and due to numerous requests from the member states, the Traditional Medicine Program of the WHO began the extensive task of reviewing the world's scientific literature of commonly used herbal medicines and publishing this information in monographs. The WHO monographs are technical reviews of the quality, safety and efficacy of commonly used herbal medicines and are intended primarily to harmonize the proper use of herbal medicines throughout the world.


Targeted therapy and pharmacogenomic programs.
The goal in administering chemotherapeutics is to develop the ability to predict the outcome of therapy in terms of response and toxicity. Technology has been developed to allow tumor profiling with measurement of protein expression, gene expression levels of markers, and even genetic polymorphisms, which may predict response to particular chemotherapeutics. The chemotherapeutics for which particular markers have been shown to predict outcome include the fluoropyrimidines and platinums. The next step is to develop clinical trials that will assess prospectively the benefits of profiling a patient's particular tumor, which should translate into an improvement in response and toxicity.


Marmoricola aurantiacus gen. nov., sp. nov., a coccoid member of the family Nocardioidaceae isolated from a marble statue.
A Gram-positive, aerobic bacterium with coccoid cells occurring singly, in pairs and in clusters was isolated from the surface of a marble statue. The peptidoglycan contain LL-diaminopimelic acid as diagnostic diamino acid and a single glycine residue as interpeptide bridge (type A3 gamma). The major menaquinone is MK-8(H4). The cellular fatty acid pattern consists of straight chain saturated and monounsaturated components and 10-methyl octadecanoic (tuberculostearic) acid as the only branched chain fatty acid. Phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol occur as characteristic polar lipids. The DNA G + C composition is 72 mol%. According to its phylogenetic position and 16S rDNA signature nucleotides, the organism is a member of the family Nocardioidaceae. The combination of chemotaxonomic characteristics is unique within this family and supports the description of a new genus and new species, Marmoricola aurantiacus. The type strain is strain BC 361T (= DSM 12652T).


Endocrine-dependent rat mammary tumor regression: use of a gonadotropin releasing hormone analog.
Long-term administration of [D-Leu6, des-Gly-NH210, Pro-ethylamide9]-GnRH, an analog of gonadotropin releasing hormone, caused regression of neoplastic tissue in a rat bearing a spontaneous mammary adenocarcinoma and in rats in which tumors had been induced by treatment with dimethylbenzanthracene (DMBA). During two separate treatment periods with the analog, the tumor in the single animal regressed although it had previously grown spontaneously. After a third period of growth, ovariectomy also induced regression, suggesting endocrine dependency of the tumor. These observations were confirmed in the DMBA-induced tumor system, where tumor regression in the analog-treated rats was comparable to that observed in the ovariectomized rats, and in both cases the tumor regression was significant when compared to untreated controls.


[Effect of carbacholine on miniature potentials and currents of rat skeletal muscle endplates].
In rat soleus muscle placed in solution with 13 mumol/l potassium chloride carbachol (10 mumol/l) caused a marked increase in miniature end-plate potentials frequency. Intensification of spontaneous mediator release was also observed under clamping of the membrane potential of the muscle fibre at potassium equilibrium potential level. It is concluded that carbachol presynaptic effect on spontaneous acetylcholine release is not connected with postsynaptic membrane depolarization and results from direct mimetic action on the motor nerve terminals.


Assessment of specific antibodies to F protein in serum samples from Chinese hepatitis C patients treated with interferon plus ribavarin.
The hepatitis C virus (HCV) alternate reading frame protein or F protein of the HCV 1b genotype is a double-frameshift product of the HCV core protein. In order to assess the presence of antibodies specific for F protein and their clinical relevance in sera from HCV patients, we produced recombinant F protein and core protein of the HCV 1b genotype in Escherichia coli. An enzyme-linked immunosorbent assay was developed using purified recombinant HCV core, F protein, and a 99-residue synthetic F peptide (F99). The seroprevalences of anticore, anti-F protein, and anti-F99 synthetic peptide were 95%, 68%, and 36%, respectively, in 168 HCV patients. The prevalence of anti-F antibodies did not correlate with viral load, genotype, or alanine aminotransferase level. Interferon combination therapy induced a decline in the level of anti-F antibodies in 55 responders (P < 0.01). Thirteen responders (24%) lost their anti-F recombinant protein antibodies, and 17 (31%) lost their anti-F synthetic peptide antibodies, whereas no decrease was observed for the 17 nonresponders. These changes were significant between responders and nonresponders (P < 0.05). Meanwhile, no change was found in the anticore antibody titer of the 72 treated patients. The percentage of anti-F-protein-negative patients (15/15 [100%]) who achieved a sustained virological response (SVR) was higher than that of the anti-F-positive patients (70%) (P < 0.05). Based on these findings, HCV F protein elicits a specific antibody response other than the anticore protein response. Our data also suggest that the presence and level of anti-F antibody responses might be influenced by the treatment (interferon plus ribavirin) and associated with an SVR in Chinese hepatitis C patients.


Predictors of Renal Outcomes in Sclerotic Class Anti-Neutrophil Cytoplasmic Antibody Glomerulonephritis.
The prognostic value of the anti-neutrophil cytoplasmic antibody (ANCA) associated glomerulonephritis (GN) classification has been demonstrated in several cohorts with sclerotic class having the worst renal outcome. Relevant published data on factors predicting outcomes in sclerotic ANCA GN is limited. Sclerotic ANCA GN patients were recruited from 5 centers worldwide for this retrospective cohort study. We describe the clinical characteristics of this cohort and evaluate predictors of 1-year glomerular filtration rate (GFR) and end-stage renal disease (ESRD). Kidney function at 12 months as measured by Modification of Diet in Renal Disease estimated GFR (eGFR) was modeled by simple and multiple linear regression analyses. We used Cox proportional hazards regression modeling to evaluate ESRD-free survival. Of the 50 patients, 92% were Caucasian and 60% male with a mean age of 61 years. While 72% had renal limited disease, 82% were MPO ANCA positive. Kidney biopsies contained a median of 20 (interquartile range [IQR] 15-34) glomeruli with 96% showing moderate to severe interstitial fibrosis. Overall, 96% of patients received immunosuppressive drug therapy and 16% received plasmapheresis. Treatment response was achieved in all but 1 patient. The median (IQR) eGFR at entry was 14.5 (9-19) mL/min/1.73 m2. Over a median (IQR) follow-up of 33.5 (17-82) months, 26 patients reached ESRD. Ten patients died with 6 of the deaths occurring within the first year of diagnosis. The hazard of progression to ESRD was significantly higher in those with lower GFR at study entry (p = 0.003) and with higher degree of tubular atrophy (p = 0.043). Renal recovery is rare among sclerotic ANCA GN patients requiring dialysis at entry and 12% of patients died in the first year. Entry GFR and tubular atrophy were significant predictors of GFR at 12 months and renal survival in patients with sclerotic class ANCA GN.


Chinese mental illness narratives: controlling the spirit.
Through analysis of local level narrative themes and linguistic features and their attendant narrative 'domains' and emotive content, this study explicates the discursive forms of Chinese personal narratives about mental illness published in a psychoeducational volume in mainland China. The cultural phenomena underpinning the explicated discursive forms are considered. The study finds that the Chinese mental illness narratives emphasize the problematic nature of the illness experience for the individual concerned, as well as the importance of gaining control over the personal and social disembodiment that mental illness brings about. The language of medicine presents as a dominant linguistic feature of the texts. Narrative authors appear to respond to their illness experience by negotiating and utilizing social relationships and support structures and through active medical intercession. As such, their narratives appear deeply culturally marked despite their psychoeducational intent and institutional context of publication. The study findings bear out the call for more attention to be given to the cultural dimension when analysing illness narratives.


Centromere Biology: Transcription Goes on Stage.
Accurate chromosome segregation is a fundamental process in cell biology. During mitosis, chromosomes are segregated into daughter cells through interactions between centromeres and microtubules in the mitotic spindle. Centromere domains have evolved to nucleate formation of the kinetochore, which is essential for establishing connections between chromosomal DNA and microtubules during mitosis. Centromeres are typically formed on highly repetitive DNA that is not conserved in sequence or size among organisms and can differ substantially between individuals within the same organism. However, transcription of repetitive DNA has emerged as a highly conserved property of the centromere. Recent work has shown that both the topological effect of transcription on chromatin and the nascent noncoding RNAs contribute to multiple aspects of centromere function. In this review, we discuss the fundamental aspects of centromere transcription, i.e., its dual role in chromatin remodeling/CENP-A deposition and kinetochore assembly during mitosis, from a cell cycle perspective.


Visualization and quality assessment of the contrast transfer function estimation.
The contrast transfer function (CTF) describes an undesirable distortion of image data from a transmission electron microscope. Many users of full-featured processing packages are often new to electron microscopy and are unfamiliar with the CTF concept. Here we present a common graphical output to clearly demonstrate the CTF fit quality independent of estimation software. Separately, many software programs exist to estimate the four CTF parameters, but their results are difficult to compare across multiple runs and it is all but impossible to select the best parameters to use for further processing. A new measurement is presented based on the correlation falloff of the calculated CTF oscillations against the normalized oscillating signal of the data, called the CTF resolution. It was devised to provide a robust numerical quality metric of every CTF estimation for high-throughput screening of micrographs and to select the best parameters for each micrograph. These new CTF visualizations and quantitative measures will help users better assess the quality of their CTF parameters and provide a mechanism to choose the best CTF tool for their data.


Ablation of NF1 function in neurons induces abnormal development of cerebral cortex and reactive gliosis in the brain.
Neurofibromatosis type 1 (NF1) is a prevalent genetic disorder that affects growth properties of neural-crest-derived cell populations. In addition, approximately one-half of NF1 patients exhibit learning disabilities. To characterize NF1 function both in vitro and in vivo, we circumvent the embryonic lethality of NF1 null mouse embryos by generating a conditional mutation in the NF1 gene using Cre/loxP technology. Introduction of a Synapsin I promoter driven Cre transgenic mouse strain into the conditional NF1 background has ablated NF1 function in most differentiated neuronal populations. These mice have abnormal development of the cerebral cortex, which suggests that NF1 has an indispensable role in this aspect of CNS development. Furthermore, although they are tumor free, these mice display extensive astrogliosis in the absence of conspicuous neurodegeneration or microgliosis. These results indicate that NF1-deficient neurons are capable of inducing reactive astrogliosis via a non-cell autonomous mechanism.


Probing the "charge cluster mechanism" in amphipathic helical cationic antimicrobial peptides.
Clustering of anionic lipids away from zwitterionic ones by cationic antimicrobial agents has recently been established as a mechanism of action of natural small, flexible peptides as well as non-natural synthetic peptide mimics. One of the largest classes of antimicrobial peptides consists of peptides that form cationic amphipathic helices on membranes and whose toxic action is dependent on the formation of pores in the membrane or through the "carpet" mechanism. We have evaluated the role of anionic lipid clustering for five of these peptides, i.e., MSI-78, MSI-103, MSI-469, MSI-843, and MSI-1254, with different sequences and properties. We determined whether these amphipathic helical cationic antimicrobial peptides cluster anionic lipids from zwitterionic ones and if this property is related to the species specificity of their toxicity. All five of these peptides were capable of lipid clustering, in contrast to the well-studied amphipathic helical antimicrobial peptide, magainin 2, which does not. We ascribe this difference to the lower density of positive charges in magainin 2. Peptides that efficiently cluster anionic lipids generally have a ratio of MIC for Staphylococcus aureus to that for Escherichia coli of >1. The addition of an N-terminal octyl chain did not preclude anionic charge clustering, although the ratio of MIC for S. aureus to that for E. coli was somewhat lowered. In most Gram-positive bacteria, there is a predominance of anionic lipids in the cytoplasmic membrane. In Gram-negative bacteria, however, clustering of anionic lipids away from zwitterionic ones is emerging as an important contributing mechanism of bacterial toxicity for some antimicrobial agents.


CKD and cardiovascular disease in the Atherosclerosis Risk in Communities (ARIC) study: interactions with age, sex, and race.
Estimated glomerular filtration rate (eGFR) and albuminuria are central for diagnosis, staging, and risk evaluation in chronic kidney disease (CKD). Universal thresholds regardless of age, sex, and race are recommended, but relatively little is known about how these demographic factors alter the relationship of eGFR and albuminuria to cardiovascular outcomes. Observational cohort study. 11,060 whites and blacks aged 52-75 years in the Atherosclerosis Risk in Communities (ARIC) Study with median follow-up of 11.2 years. eGFR by the CKD-EPI (CKD Epidemiology Collaboration) creatinine equation (reference, 95 mL/min/1.73 m(2)) and urinary albumin-creatinine ratio (ACR; reference, 5 mg/g). Cardiovascular events (coronary disease, stroke, and heart failure) and all-cause mortality. Adjusted HRs associated with eGFR and ACR in subgroups according to age, sex, and race. Cardiovascular risk significantly increased at eGFR <70 mL/min/1.73 m(2) in all subgroups according to age (<65 vs ≥65 years), sex, and race (P for interaction >0.2 for these subgroups; eg, at eGFR of 30 mL/min/1.73 m(2), the adjusted HR was 2.19 [95% CI, 1.10-4.35] at age 52-64 years vs 2.23 [95% CI, 1.33-3.72] at age 65-75 years). Results were similar for mortality. Log(ACR) was associated linearly with cardiovascular risk without threshold effects in all subgroups, with some quantitative interactions. HRs according to ACR tended to be lower in men versus women (eg, at ACR of 40 mg/g, 1.18 [95% CI, 0.98-1.41] vs 1.77 [95% CI, 1.45-2.15]) and in the older versus younger population (1.24 [95% CI, 1.04-1.49] vs 1.73 [95% CI, 1.42-2.12]; P for interaction <0.01 for sex and age). Less evident interactions were observed for mortality. Single measurement of eGFR with creatinine and ACR and relatively narrow age range. The associations of eGFR and ACR with cardiovascular events were largely similar, with some quantitative interactions, in age, sex, and racial subgroups, generally supporting universal thresholds of GFR and ACR for CKD definition/staging.


Patent Macracanthorhynchus ingens Infection in a 17-Month-Old Child, Ohio.
Limited data exist on human Macracanthorhynchus infections. We report an asymptomatic 17-month-old who passed eggs and an adult Macracanthorhynchus ingens worm, indicating parasite maturation and reproduction. Macracanthorhynchus ingens may have a greater capacity to mature in humans versus Macracanthorhynchus hirudinaceus.


Effects of social rearing conditions on conditioned suppression in rats.
Twenty-two rats were reared in standard conditions during the first two months of their life. Then the animals were divided into two groups exposed to different rearing conditions. Twelve animals (Group SO) were housed socially, six animals per cage, and for three weeks they were subjected to sensory stimulation in an enriched environment. The other ten subjects were kept individually (Group IN); one rat per mesh cage, in conditions of relatively impoverished sensory stimulation. In both groups the training of the conditioned emotional response (CER) was performed when animals were three months old. In contrast to IN subjects, the rats subjected to permanent social contacts and reared in the enriched environment (Group SO) revealed almost equally low instrumental response rates in trials with the conditioned stimulus (CS) paired with nociceptive foot-shock (US), and in periods when no CS and/or US were applied. The results suggested that early exposure to an enriched environment caused a later decrease of the animals' capability to differentiate between the aversive CS and cues of the experimental context. This cognitive impairment was probably a secondary effect of fear generalized to the entire experimental situation.


French field experience with buprenorphine.
In most European countries, methadone treatment is provided to only 20-30% of opiate abusers who need treatment due to regulations and concerns about safety. To address this need in France, all registered medical doctors since 1995 have been allowed to prescribe buprenorphine (BUP) without any special education or licensing. This led to treating approximately 65,000 patients per year with BUP, about ten times more than with more restrictive methadone policies. French physician compensation mechanisms, pharmacy services, and medical insurance funding all minimized barriers to BUP treatment. About 20% of all physicians in France are using BUP to treat about half of the estimated 150,000 problem heroin users. Daily supervised dosing by a pharmacist for the first six months resulted in significantly better treatment retention (80% vs 46%) and lower heroin use. Intravenous diversion of BUP may occur in up to 20% of BUP patients and has led to various infections and relatively rare overdoses in combination with sedatives. Opiate overdose deaths have declined substantially (by 79%) since BUP was introduced in 1995. Newborn opiate withdrawal in mothers treated with buprenorphine compared to methadone was reported to be less frequent, less severe, and of shorter duration. Although some of the public health benefits seen during the time of buprenorphine expansion in France might be contingent upon characteristics of the French health and social services system, the French model raises questions about the value of tight regulations on prescribing BUP imposed by many countries throughout the world.