Amplification of signaling events in bacteria.
Bacteria respond to extremely shallow chemical gradients by modifying their motility in a process called chemotaxis. This chemotactic response is characterized by high sensitivity to small concentration differences, which extends over a large range of concentrations. This combination of high signal gain and large dynamic range results from both a memory of past events and the ability to amplify small differences in signal between the memory and the current environment. Dahlquist describes the signaling mechanism used by bacteria to regulate the flagellar motor and the places in this pathway where signal amplification may occur.


Genotyping of Plasmodium falciparum infections by PCR: a comparative multicentre study.
Genetic diversity of malaria parasites represents a major issue in understanding several aspects of malaria infection and disease. Genotyping of Plasmodium falciparum infections with polymerase chain reaction (PCR)-based methods has therefore been introduced in epidemiological studies. Polymorphic regions of the msp1, msp2 and glurp genes are the most frequently used markers for genotyping, but methods may differ. A multicentre study was therefore conducted to evaluate the comparability of results from different laboratories when the same samples were analysed. Analyses of laboratory-cloned lines revealed high specificity but varying sensitivity. Detection of low-density clones was hampered in multiclonal infections. Analyses of isolates from Tanzania and Papua New Guinea revealed similar positivity rates with the same allelic types identified. The number of alleles detected per isolate, however, varied systematically between the laboratories especially at high parasite densities. When the analyses were repeated within the laboratories, high agreement was found in getting positive or negative results but with a random variation in the number of alleles detected. The msp2 locus appeared to be the most informative single marker for analyses of multiplicity of infection. Genotyping by PCR is a powerful tool for studies on genetic diversity of P. falciparum but this study has revealed limitations in comparing results on multiplicity of infection derived from different laboratories and emphasizes the need for highly standardized laboratory protocols.


Multifocal ERG recordings under visual control of the stimulated fundus in mice.
Therapeutic approaches to retinal disease require a continuous monitoring of functional improvement over lesion areas that sometimes cannot be shown in full-field ERG. The aim of this study was to assess the usefulness of multifocal electroretinograms (mfERGs) under visual control using scanning laser ophthalmoscopy (SLO) for evaluation of local retinopathy in mice. mfERGs were optimized for recordings in C57BL/6 mice by varying dark steps between each stimuli, background intensity, and the numbers of hexagons. Local retinopathy was induced by argon laser photocoagulation with different spot sizes and retinal irradiances. mfERG recordings were performed before, and 10 days and 4 weeks after laser treatment. In each recording, the central hexagon was positioned on the optic nerve head visualized by SLO images. The amplitudes of the P1 response components were analyzed as a function of retinal location. The mfERG amplitudes depended on stimulus condition. The P1 amplitudes increased with increasing number of dark frames in the m-sequence and with decreasing number of hexagons. A stimulus with 19 hexagons and four dark frames was chosen because substantial response amplitudes could be achieved while preserving sufficient spatial resolution. In the untreated eyes, the response to the central hexagon, stimulating the optic nerve head, was smaller than those to the surrounding hexagons. The responses to hexagons stimulating photocoagulated areas were reduced compared with the responses of surrounding areas. The amplitude reduction was more pronounced when the coagulated areas were larger and when higher energies were used. Areas with decreased sensitivities to light stimulation (either the optic nerve head or damaged retinal areas) can be detected and correlated with the retinal images and in the mfERG responses. We demonstrate that the mfERG technique is able to reproducibly detect the functional consequences of a local treatment.


A technology-enabled adherence enhancement system for people with bipolar disorder: results from a feasibility and patient acceptance analysis.
As poor medication adherence is common in bipolar disorder (BD), technology-assisted approaches may help to monitor and enhance adherence. This study evaluated preliminary feasibility, patient satisfaction and effects on adherence, BD knowledge, and BD symptoms associated with the use of a multicomponent technology-assisted adherence enhancement system. This prospective study tested the system in five BD patients over a 15-day period. System components included: 1) an automated pill cap with remote monitoring sensor; 2) a multimedia adherence enhancement program; and 3) a treatment incentive program. This study evaluated system usability, patient satisfaction and effects on adherence (Morisky scale), knowledge (treatment knowledge test [TKT]), and symptoms (internal state scale [ISS]). Mean age of the sample was 62 years, 4/5 (80%) Caucasian, and 4/5 (80%) single/divorced or widowed. Most participants (4/5, 80%) were on a single BD medication. Participants had BD for an average of 21 years. Challenges included attaching the pill sensor to standard pharmacy bottles for individuals using very large pill containers or those with multiday pill boxes. Three of five (60%) individuals completed the full 15-day period. Usability scores were high overall. Mean Morisky scores improved. Means on all four subscales of the ISS were all in the direction of improvement. On the TKT, there was a 40% increase in mean scores. A multicomponent technology-assisted BD adherence enhancement system is feasible. Challenges include accommodating multiple types of pill containers and monitoring multiple drugs simultaneously. The system can also generate adherence information that is potentially useful for treatment planning.


Bronchodilatory effects of S-isopetasin, an antimuscarinic sesquiterpene of Petasites formosanus, on obstructive airway hyperresponsiveness.
In the presence of neostigmine (0.1 microM), S-isopetasin competitively antagonized cumulative acetylcholine-induced contractions in guinea pig trachealis, because the slope [1.18+/-0.15 (n=6)] of Schild's plot did not significantly differ from unity. The pA2 value of S-isopetasin was calculated to be 4.62+/-0.05 (n=18). The receptor binding assay for muscarinic receptors of cultured human tracheal smooth muscle cells (HTSMCs) was performed using [3H]-N-methylscopolamine ([3H]-NMS). Saturation binding assays were carried out with [3H]-NMS in the presence (non-specific binding) and absence (total binding) of atropine (1 microM). Analysis of the Scatchard plot (y=0.247-1.306x, r2=0.95) revealed that the muscarinic receptor binding sites in cultured HTSMCs constituted a single population (n(H)=1.00). The equilibrium dissociation constant (Kd) and the maximal receptor density (B(max)) for [3H]-NMS binding were 766 pM and 0.189 pmol/mg of protein, respectively. The -logIC50 values of S-isopetasin, methoctramine, and 1,1-Dimethyl-4-diphenylacetoxypiperidinium iodide (4-DAMP) for displacing 0.4 nM [3H]-NMS-specific binding were 5.05, 6.25, and 8.56, respectively, which suggests that [3H]-NMS binding is predominantly on muscarinic M3 receptors of cultured HTSMCs. The inhibitory effects of S-isopetasin on enhanced pause (P(enh)) value were similar to that of ipratropium bromide, a reference drug. The duration of action of S-isopetasin (20 microM), also similar to that of ipratropium bromide (20 microM), was 3 h. In contrast to ipratropium bromide, which non-selectively acts on muscarinic receptors, S-isopetasin preferentially acts on muscarinic M3 receptors. In conclusion, S-isopetasin may be beneficial as a bronchodilator in the treatment of chronic obstructive pulmonary disease and asthma exacerbations.


Nettle as a distinct Bronze Age textile plant.
It is generally assumed that the production of plant fibre textiles in ancient Europe, especially woven textiles for clothing, was closely linked to the development of agriculture through the use of cultivated textile plants (flax, hemp). Here we present a new investigation of the 2800 year old Lusehøj Bronze Age Textile from Voldtofte, Denmark, which challenges this assumption. We show that the textile is made of imported nettle, most probably from the Kärnten-Steiermark region, an area which at the time had an otherwise established flax production. Our results thus suggest that the production of woven plant fibre textiles in Bronze Age Europe was based not only on cultivated textile plants but also on the targeted exploitation of wild plants. The Lusehøj find points to a hitherto unrecognized role of nettle as an important textile plant and suggests the need for a re-evaluation of textile production resource management in prehistoric Europe.


Optical trapping and binding of particles in an optofluidic stable Fabry-Pérot resonator with single-sided injection.
In this article, microparticles are manipulated inside an optofluidic Fabry-Pérot cylindrical cavity embedding a fluidic capillary tube, taking advantage of field enhancement and multiple reflections within the optically-resonant cavity. This enables trapping of suspended particles with single-side injection of light and with low optical power. A Hermite-Gaussian standing wave is developed inside the cavity, forming trapping spots at the locations of the electromagnetic field maxima with a strong intensity gradient. The particles get arranged in a pattern related to the mechanism affecting them: either optical trapping or optical binding. This is proven to eventually translate into either an axial one dimensional (1D) particle array or a cluster of particles. Numerical simulations are performed to model the field distributions inside the cavity allowing a behavioral understanding of the phenomena involved in each case.


TEMPERATURE-SENSITIVE REPRESSION OF STAPHYLOCOCCAL PENICILLINASE.
In eight highly inducible strains of Staphylococcus aureus repression of the formation of penicillinase was temperature-sensitive under conditions suggesting direct thermal inactivation of the repressor. Restoration of repression required protein synthesis. These strains were resistant to benzylpenicillin and to many other antibiotics. One auxotrophic mutant had gredtly reduced temperature sensitivity but was still normally inducible. Six strains were relatively poorly inducible, exhibited a proportionately smaller increase in enzyme after exposure to elevated temperature, and were sensitive to antibiotics other than benzylpenicillin. Temperature sensitivity may be a useful character in studies of the physiology and genetics of the repression of staphylococcal penicillinase.


The association of ADHD symptoms and reading acquisition during elementary school years.
The present longitudinal study aimed to investigate the influence of ADHD symptoms on reading development in elementary schoolchildren. To this end, repeated assessments of ADHD symptoms (teacher ratings of inattention, hyperactivity, and impulsivity) and reading achievement (standardized tests of decoding speed and text comprehension) were examined in a sample comprising 2,014 elementary schoolchildren at the end of Grades 1, 2, 3, respectively, and in the middle of Grade 4. Latent change score models revealed that the level of ADHD symptoms was associated with lower levels and less growth in decoding speed and text comprehension. Furthermore, individual differences in changes in ADHD symptoms and reading performance were negatively associated. Together, these results indicate commonalities in the development of ADHD symptomatology and reading achievement throughout elementary school. (PsycINFO Database Record


Initial development of patient-reported instrument assessing harm, efficacy, and misuse of long-term opioid therapy.
Guidelines on long-term opioid therapy recommend frequent reassessment of harm, efficacy, and misuse of these potentially harmful and sometimes ineffective medications. In primary care, there is a need for a brief, patient-reported instrument. This report details the initial steps in the development of such an instrument. An interdisciplinary team of clinician-scientists performed four discrete steps in this study: (1) conceptualization of the purpose and function of the instrument, (2) assembly of an item pool, (3) expert rating on which items were most important to include in the instrument, and (4) modification of expert-selected items based on a reading level check and cognitive interviews with patients. A diverse panel of 47 subject matter experts was presented with 69 items to rate on a 1-9 scale in terms of importance for inclusion in the instrument. The panel highly rated 37 items: 8 related to harm, 4 related to efficacy, and 25 related to misuse. These 37 items were then tested for patient comprehension and modified as needed. Next steps in development will include further item reduction, testing against a gold standard, and assessment of the instrument's effect on clinical outcomes.


Gamma Knife radiosurgery in the treatment of abducens nerve schwannomas: a retrospective study.
OBJECTIVE Of the intracranial schwannomas, those arising from the vestibular nerves are the most common. Abducens nerve (AN) schwannomas are very rare, and there is limited literature on their optimal management. Therapeutic options include surgery and/or stereotactic radiosurgery. The aim of this study was to evaluate the role of Gamma Knife radiosurgery (GKRS) in these sixth cranial nerve (CN) schwannomas. METHODS The authors performed a retrospective analysis of patients who had undergone GKRS for intracranial tumors at their institute in the period from 2003 to 2010. Inclusion criteria were as follows: isolated AN paresis on presentation, a lesion along the course of the sixth CN, and imaging features characteristic of a schwannoma. Patients with other CN deficits and neurofibromatosis Type 2 were excluded. Symptomatic improvement was defined as the resolution of or an improvement in diplopia noted on a subjective basis or as an improvement in lateral eyeball excursion noted objectively on follow-up. A reduction in tumor volume by at least 20%, as noted by comparing the pre- and post-GKRS images, was deemed significant. RESULTS Six patients with a mean age of 37.1 years (range 17-55 years) underwent primary GKRS. There were 2 prepontine cistern, 3 cavernous sinus, and 1 cisterno-cavernous tumor. The mean duration of symptoms was 6.1 months (range 3-12 months). The mean tumor volume was 3.3 cm3 (range 1.5-4.8 cm3). The mean tumor margin radiation dose was 12.5 Gy (range 12-14 Gy), while the median margin dose was 12 Gy (50% isodose line). The median number of isocenters used was 5 (range 4-8). The brainstem received an average 8.35-Gy radiation dosage (range 5.5-11 Gy). The mean follow-up duration was 44.3 months (range 24-78 months). Symptoms remained stable in 1 patient, improved in 3, and resolved in 2 (total improvement 83%). Magnetic resonance imaging at the last follow-up showed a stable tumor size in 3 patients (50%) and a reduction in the other 3. Thus, the tumor control rate achieved was 100%. No new CN deficits were noted. CONCLUSIONS Abducens nerve schwannomas are rare intracranial tumors. They can be cavernous, cisternal, or cisterno-cavernous in location. Excellent tumor control rates and symptomatic improvement can be achieved with GKRS, which appears to be a safe and effective, minimally invasive modality for the treatment of such lesions. Therefore, it is reasonable to consider GKRS as the initial treatment of choice for this rare pathology. Long-term follow-up will be essential for further recommendations.


In-vitro viral suppressive capacity correlates with immune checkpoint marker expression on peripheral CD8+ T cells in treated HIV-positive patients.
To determine whether viral suppressive capacity (VSC) of CD8+ T cells can be boosted by stimulation with HIV-1 peptides and whether the ability to control HIV-1 replication correlates with immunological (cytokine production and CD8+ T-cell phenotype) and viral reservoir measures (total HIV-1 DNA and cell-associated RNA) in well treated HIV-infected chronic progressors. We compared VSC of peripheral CD8+ T cells to cytokine production profile in response to peptide stimulation, detailed phenotype (17-color flow-cytometry), reservoir size (total HIV-1 DNA), basal viral transcription (unspliced cell-associated RNA) and inducible viral transcription (tat/rev induced limiting dilution assay) in 36 HIV+ patients on cART and six healthy donors. We found that the VSC of CD8+ T cells can be increased by prior stimulation with a pool of consensus HIV-1 gag peptides in a significant proportion of progressor patients. We also found that VSC after peptide stimulation was correlated with higher expression of immune checkpoint markers on subsets of terminally differentiated effector memory (TEMRA) CD8 T cells as well as with production of IFN-γ, TNF-α and IL-10. We did not find a correlation between VSC and viral reservoir measures. These results add to a small body of evidence that the capacity of CD8+ T cells to suppress viral replication is increased after stimulation with HIV-1 peptides. Interestingly, this VSC was correlated with expression of immune checkpoint markers, which are generally considered to be markers of exhaustion. Our findings may guide further investigations into immune phenotypes correlated with viral suppression.


Description of the immatures of the ant, Myrmelachista catharinae.
The Neotropical ant genus Myrmelachista Roger comprises 69 described species and subspecies, and still is a poorly studied group. Larvae play a paramount role in colony nutrition in social hymenopterans and bear considerable value in the reconstruction of group phylogenies, however, they are generally neglected. Larvae of different instars of Myrmelachista catharinae Mayr (Hymenoptera: Formicidae) are herein described in detail by light and scanning electron microscopy. The number of larval instars was estimated as three based on the frequency distribution of maximum head capsule widths. The described larvae confirmed some traits typical of the genus: general shape of body and mandibles, general aspect and distribution of body hairs, and the number of sensilla on the palps and galea. Differently from other Myrmelachista larvae previously described, M. catharinae presented two distinct kinds of second instars, some additional types of body hairs, different number of antennal sensilla, and a distinct labrum shape. M. catharinae presented ten pairs of spiracles, which is the first record for this genus.


Convergent origination of a Drosophila-like dosage compensation mechanism in a reptile lineage.
Sex chromosomes differentiated from different ancestral autosomes in various vertebrate lineages. Here, we trace the functional evolution of the XY Chromosomes of the green anole lizard (Anolis carolinensis), on the basis of extensive high-throughput genome, transcriptome and histone modification sequencing data and revisit dosage compensation evolution in representative mammals and birds with substantial new expression data. Our analyses show that Anolis sex chromosomes represent an ancient XY system that originated at least ≈160 million years ago in the ancestor of Iguania lizards, shortly after the separation from the snake lineage. The age of this system approximately coincides with the ages of the avian and two mammalian sex chromosomes systems. To compensate for the almost complete Y Chromosome degeneration, X-linked genes have become twofold up-regulated, restoring ancestral expression levels. The highly efficient dosage compensation mechanism of Anolis represents the only vertebrate case identified so far to fully support Ohno's original dosage compensation hypothesis. Further analyses reveal that X up-regulation occurs only in males and is mediated by a male-specific chromatin machinery that leads to global hyperacetylation of histone H4 at lysine 16 specifically on the X Chromosome. The green anole dosage compensation mechanism is highly reminiscent of that of the fruit fly, Drosophila melanogaster Altogether, our work unveils the convergent emergence of a Drosophila-like dosage compensation mechanism in an ancient reptilian sex chromosome system and highlights that the evolutionary pressures imposed by sex chromosome dosage reductions in different amniotes were resolved in fundamentally different ways.


The effect of changing gonadotropin-releasing hormone pulse frequency on puberty.
We have examined the effect of using different pulse frequencies of exogenous GnRH to induce puberty and the time relationship among LH, FSH, sex steroids, and GH in these individuals. Five girls and three boys with delayed puberty received exogenous GnRH at either 3-h frequency (slow) or every 45 min (fast). Treatment was initially given overnight and increased to 24 h when breast stage 3 in girls or testicular volume of 10 mL in boys was attained. Twenty-four-hour gonadotropin profiles were performed after 5 days, 1 month, 3 months, 6 months, and 1 yr of treatment. Temporal relationships among LH, FSH, and estradiol; LH and testosterone; GH and estradiol; and GH and testosterone were examined by cross-correlation. There was no difference in the rate of pubertal progress between the groups. Mean serum gonadotropin and sex steroid levels did not differ. LH was correlated with estradiol for both groups at 240 min (slow group, r = 0.54; fast group, r = 0.50). Estradiol correlated with LH at 300 min in the slow group (r = -0.41) and 200 min in the fast group (r = -0.37). FSH correlated with estradiol at 140 min in the slow group (r = 0.62) and 160 min in the fast group (r = 0.50). A rise in estradiol occurred 140-160 min after a rise in FSH and 240 min after a rise in LH. A rise in estradiol was followed 200-300 min later by a fall in LH. LH was correlated with testosterone at 60 min in the slow group (r = 0.73) and at 40 min in the fast group (r = 0.55). Testosterone correlated with LH at 420 min in the slow (r = -0.67) and 460 min in the fast group (r = -0.40). A rise in LH was followed 40-60 min later by a rise in testosterone. A rise in testosterone was followed by fall in LH 420-460 min later. GH correlated with estradiol at 320 min in the slow group (r = 0.37) and 380 min in the fast group (r = 0.38). A rise in GH was followed, after 320-380 min, by a rise in estradiol. There was a correlation between GH and testosterone in the slow group after 280 min (r = 0.44). A rise in GH was followed by a rise in testosterone after 280 min. The pituitary-gonadal axis is sufficiently robust to allow puberty to progress with different fixed pulse frequencies. There is a temporal relationship among LH, FSH, and estradiol secretion and between LH and testosterone secretion. We have demonstrated the feedback effect of sex steroids at the level of the pituitary and the time course of the effect of GH on gonadal function.


Development of a real-time PCR assay for quantitative detection of Encephalitozoon intestinalis DNA.
A new real-time PCR assay for quantitation of Encephalitozoon intestinalis DNA was developed which used a TaqMan fluorescent probe for specific detection. Serial dilutions of E. intestinalis spore suspensions obtained from tissue culture were used as external standards. The detection limit of the technique was 20 spores per ml, with a good interassay reproducibility (coefficient of variation of 7.1% for the suspension containing 20 spores/ml, 5.0% for the suspension containing 75 spores/ml and below 3.5% for higher concentrations). Quantitative detection of E. intestinalis DNA was similar whether the serial dilutions of spores were made in distilled water or in a stool suspension, allowing the use of the assay for stool specimens. The assay was then applied to 14 clinical specimens from 8 immunocompromised patients with proven E. intestinalis infection. The quantitation of the parasitic burden was achieved in stools, blood, urine, tissue biopsies, and bronchopulmonary specimens. The highest parasitic burdens were noted in stools, urine, and bronchopulmonary specimens, reaching 10(5) to 10(6) spores/g or ml. Dissemination of the infection was also evidenced in some patients by demonstration of E. intestinalis DNA in blood and serum. We conclude that real-time PCR is a valuable tool for quantitation of E. intestinalis burden in clinical specimens.


'Hotspots' for aggression in licensed drinking venues.
In order to better understand the social context of barroom aggression, the aim was to identify common locations ('hotspots') for aggression in bars and examine the association of hotspots with aggression severity and environmental characteristics. Aggression hotspots were identified using narrative descriptions and data recorded on premises' floor plans for 1057 incidents of aggression collected in the Safer Bars evaluation. Hierarchical Linear Modelling was used to identify bar-level and night-level characteristics associated with each hotspot. The most common location for aggression was the dance floor (20.0% of incidents) or near the dance floor (11.5%), followed by near the serving bar (15.7%), at tables (13.1%), aisles, hallways and other areas of movement (6.2%), entrance (4.5%) and the pool playing area (4.1%). Hotspots were predicted mainly by bar-level characteristics, with dance floor aggression associated with crowded bars, a high proportion of female and young patrons, lots of sexual activity, a large number of patrons and staff, security staff present, better monitoring and coordination by staff, and people hanging around at closing. Incidents at tables and pool tables tended to occur in bars with the opposite characteristics. Nightly variations in patron intoxication and rowdiness were associated with aggression at tables while variations in crowding and sexual activity were associated with aggression in areas of movement. Incidents outside tended to be more severe. Each aggression location and their associated environments have somewhat different implications for staff training, premises design, policy and prevention.


Construction and function of a highly efficient supramolecular luminescent system.
Aggregation-induced emission (AIE) provides a new way of achieving highly efficient luminescent materials. In this contribution, the self-assembly behavior, molecular stacking structure and photophysical properties of two polymorphs of a supramolecular co-crystal (C1 and C2) are investigated. The block-like crystal C1, packed in segregated stacking with strong π-π interactions between the H and G molecules, shows weak green emission with a low efficiency (ΦF) of 2%. In comparison, the needle-like crystal C2, packed in segregated stacking with no obviously strong intermolecular interactions, shows bright yellow emission. More importantly, C1 exhibits mechanochromic behavior.


Anergic T cells as suppressor cells in vitro.
T cell-mediated suppression is an established phenomenon, but its underlying mechanisms are obscure. An in vitro system was used to test the possibility that anergic T cells can act as specific suppressor cells. Anergic human T cells caused inhibition of antigen-specific and allospecific T cell proliferation. In order for the inhibition to occur, the anergic T cells had to be specific for the same antigen-presenting cells (APCs) as the T cells that were suppressed. The mechanism of this suppression appears to be competition for the APC surface and for locally produced interleukin-2.


The serum angiotensin-converting enzyme and angiotensin II response to altered posture and acute exercise, and the influence of ACE genotype.
The deletion (D) rather than insertion (I) allele of the angiotensin-converting enzyme (ACE) gene is associated with greater ACE activity. We examined: (1) the influence of posture change (recumbent to seated) and acute exercise on serum ACE and angiotensin II (Ang II) activity; (2) the relationship between ACE and Ang II levels; and (3) the influence of ACE genotype on changes in ACE and Ang II levels with posture and exercise. Recreationally active young male Caucasians (10 each of II, ID and DD genotypes) rested for 35 min supine then 15 min upright, took 20 min bicycle ergometric exercise at 70% maximum oxygen uptake, then rested for 40 min. Samples were taken throughout for ACE activity and Ang II levels. Supine ACE levels were dependent upon ACE genotype [24.8 (5.7), 26.9 (4.5), 45.5 (6.4) nmol His-Leu ml(-1) min(-1); II, ID, DD, respectively; P<0.00005] and thereafter. ACE activity rose with assumption of a seated posture [from 32.4 (10.9) nmol His-Leu ml(-1) min(-1) to 35.0 (11.5) nmol His-Leu ml(-1) min(-1), P<0.00001], the absolute rise being independent of genotype [3.22 (1.92), 1.6 (1.6), 2.4 (2.3) nmol His-Leu ml(-1) min(-1); II, ID, DD; P=0.22], unlike percentage change [12.8 (6.8), 5.6 (5.5), 5.3 (5.0)%; II, ID, DD; P<0.01, and P=0.004 for II vs presence of the D allele]. A further genotype-independent rise occurred with exercise [+2.9 (3.7) units, P<0.0003]. An associated rise in Ang II levels [30.3 (15.9), or 2587.9 (489.76)%, P<0.00001] was independent of ACE genotype or activity. Upright posture increases ACE activity, and this may be influenced by ACE genotype. ACE activity and Ang II levels rise independently with exercise in a non-genotype-dependent fashion.


Complete activation of thyroid hormone receptor β by T3 is essential for normal cochlear function and morphology in mice.
Thyroid hormones (THs) regulate many developmental processes, including the developmental onset of cochlear differentiation and function. TH action is mediated mostly by triiodothyronine (T3) bound to thyroid hormone nuclear receptors (TRs). At positive regulated genes and in the absence of THs, nuclear co-repressors are bound to TRs and decrease basal transcription rate. Ligand (T(3)) binding results in the dissociation of co-repressors and the recruitment of co-activators to the complex, which results in full transcriptional activation. We measured cochlear function in two knock-in mouse models: TRβ(E457A/E457A), with the TRβ co-activator binding surface (AF-2) disrupted to prevent co-activator binding; and TRβ(Δ337T/Δ337T), which is unable to bind T(3). Cochlear morphology and function were analyzed in 10-week-old normal and mutated mice. Cochlear function was determined by measuring auditory brainstem responses, cochlear tuning and compound action potential (CAP) thresholds. All TRβ(Δ337T/Δ337T) and 85% of the TRβ(E457A/E457A) mice presented elevated CAP thresholds (P < 0.05 or less). Five percent of the TRβ(E457A/E457A) mice presented normal CAP thresholds with broadened cochlear tuning. TRβ(E457A/E457A) and TRβ(Δ337T/Δ337T) presented developmental defects that led to a decreased width (P < 0.01) and an increased thickness (P<0.01) of the tectorial membrane. In addition, TRβ(Δ337T/Δ337T) animals showed an increased tectorial membrane area (P<0.01). Both mutations were deleterious to tectorial membrane development and led to important alterations in cochlear morphology and loss of cochlear function.


Bacterial cell division protein FtsZ assembles into protofilament sheets and minirings, structural homologs of tubulin polymers.
The bacterial cell division protein FtsZ is a homolog of tubulin, but it has not been determined whether FtsZ polymers are structurally related to the microtubule lattice. In the present study, we have obtained high-resolution electron micrographs of two FtsZ polymers that show remarkable similarity to tubulin polymers. The first is a two-dimensional sheet of protofilaments with a lattice very similar to that of the microtubule wall. The second is a miniring, consisting of a single protofilament in a sharply curved, planar conformation. FtsZ minirings are very similar to tubulin rings that are formed upon disassembly of microtubules but are about half the diameter. This suggests that the curved conformation occurs at every FtsZ subunit, but in tubulin rings the conformation occurs at either beta- or alpha-tubulin subunits but not both. We conclude that the functional polymer of FtsZ in bacterial cell division is a long thin sheet of protofilaments. There is sufficient FtsZ in Escherichia coli to form a protofilament that encircles the cell 20 times. The similarity of polymers formed by FtsZ and tubulin implies that the protofilament sheet is an ancient cytoskeletal system, originally functioning in bacterial cell division and later modified to make microtubules.


A novel molecular approach to assess mating success of sterile Ceratitis capitata (Diptera: Tephritidae) males in sterile insect technique programs.
Areawide sterile insect technique (SIT) programs against Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), are increasingly implemented worldwide. A key issue in SIT is to assess mating success of released sterile males, which could be currently estimated by egg hatchability and by stored sperm head measurements. We report here on a novel molecular approach that would allow detecting the presence of Mediterranean fruit fly sterile male sperm in the female spermathecae under field conditions, as a precise marker to assess mating performance. The simplicity (only two polymerase chain reactions) and reliability of this method, jointly with the capability to detect Vienna sperm in wild Mediterranean fruit fly maintained in monitoring traps for 7 d under field conditions, suggest that it could be an efficient tool when coupled with areawide SIT programs.


betaIV spectrin, a new spectrin localized at axon initial segments and nodes of ranvier in the central and peripheral nervous system.
We report the identification of betaIV spectrin, a novel spectrin isolated as an interactor of the receptor tyrosine phosphatase-like protein ICA512. The betaIV spectrin gene is located on human and mouse chromosomes 19q13.13 and 7b2, respectively. Alternative splicing of betaIV spectrin generates at least four distinct isoforms, numbered betaIVSigma1-betaIVSigma4 spectrin. The longest isoform (betaIVSigma1 spectrin) includes an actin-binding domain, followed by 17 spectrin repeats, a specific domain in which the amino acid sequence ERQES is repeated four times, several putative SH3-binding sites and a pleckstrin homology domain. betaIVSigma2 and betaIVSigma3 spectrin encompass the NH(2)- and COOH-terminal halves of betaIVSigma1 spectrin, respectively, while betaIVSigma4 spectrin lacks the ERQES and the pleckstrin homology domain. Northern blots revealed an abundant expression of betaIV spectrin transcripts in brain and pancreatic islets. By immunoblotting, betaIVSigma1 spectrin is recognized as a protein of 250 kD. Anti-betaIV spectrin antibodies also react with two additional isoforms of 160 and 140 kD. These isoforms differ from betaIVSigma1 spectrin in terms of their distribution on subcellular fractionation, detergent extractability, and phosphorylation. In islets, the immunoreactivity for betaIV spectrin is more prominent in alpha than in beta cells. In brain, betaIV spectrin is enriched in myelinated neurons, where it colocalizes with ankyrin(G) 480/270-kD at axon initial segments and nodes of Ranvier. Likewise, betaIV spectrin is concentrated at the nodes of Ranvier in the rat sciatic nerve. In the rat hippocampus, betaIVSigma1 spectrin is detectable from embryonic day 19, concomitantly with the appearance of immunoreactivity at the initial segments. Thus, we suggest that betaIVSigma1 spectrin interacts with ankyrin(G) 480/270-kD and participates in the clustering of voltage-gated Na(+) channels and cell-adhesion molecules at initial segments and nodes of Ranvier.


Enhanced expression of angiopoietin-2 and the Tie2 receptor but not angiopoietin-1 or the Tie1 receptor in a rat model of myocardial infarction.
Modulation of Tie2 receptor activity by angiopoietin ligands is crucial for angiogenesis, blood vessel maturation, and vascular endothelium integrity. The role of the angiopoietin (Ang) and Tie system in myocardial infarction is not well understood. To investigate the participation of the Ang/Tie in myocardial infarction, adult Sprague-Dawley rats with ligation of the left anterior descending coronary artery to induce myocardial infarction were studied. Ang1, Ang2, Tie1, and Tie2 were measured immediately after ligation of the coronary artery, and at 6 h, 1 and 3 days, and 1, 2, 3 and 4 weeks after ligation by Northern blotting, Western blotting, and immunohistochemical staining. Ang2 mRNA significantly increased from 2 weeks (2.1-fold) to 4 weeks (2.9-fold) after the infarction in the left ventricular free wall. Tie2 mRNA increased significantly from 1 week (2.1-fold) to 4 weeks (3.8-fold) after the infarction. Ang2 protein also significantly increased from 3 days (1.9-fold) to 4 weeks (3-fold) after the infarction in the left ventricular free wall. Tie2 protein increased 2.4-fold at 3 weeks and 2.8-fold at 4 weeks after the infarction. Neither Ang1 nor Tie1 mRNA or protein showed any significant change at any time point after the infarction. The ratio of Ang2/Ang1 mRNA and protein in the study group was higher than that in the control group. Ang2 and Tie2 expression in nonischemic myocardium showed no significant change. Immunohistochemical study also showed increased immunoreactivity of Ang2 and Tie2 at the infarct border. In conclusion, Ang2 and Tie2 expressions significantly increased both spatial and temporal patterns after myocardial infarction in the rat ventricular myocardium, while Ang1 and Tie1 receptor expression did not.


Female UK Army Service personnel are at greater risk of work-related morbidity on return to duty postpartum.
The 2016 Interim Report on the Health Risks to Women in Ground Close Combat Roles highlighted an increased risk of skeletal injury and significant physiological changes, including increased ligament laxity and decreased bone mineral content, during the postpartum period. The report called for further research and a re-evaluation of postpartum policy to optimise the return of female Service personnel to arduous employment. The purpose of this study was to determine whether returning to duty is at greater risk of injury and illness in the first year postpartum than they were prepregnancy. Fifty-five female UK Army Service personnel aged 18-41 years, who had given birth in the previous 4 years, completed a lifestyle questionnaire and gave written consent for a review of their medical records. The number of working days lost (WDL) due to illness, injury and combined illness and injury was obtained from medical records, for 1 year prepregnancy and 1 year postpartum. Female Service personnel returned to duty at different time-points postpartum, so data were expressed as WDL/week. WDL/week due to illness and combined illness and injury were higher postpartum compared with prepregnancy (p<0.05). WDL/week due to combined illness and injury was significantly lower prepregnancy (p<0.05) and at 0-26 weeks postpartum (p<0.05), compared with 26-52 weeks postpartum. Postpartum female UK Army Service personnel are at greater risk of illness and a combination of illness and injury in the year after giving birth, compared with prepregnancy. The study suggests female Service personnel are unprepared for the demands of full active duty in the first year postpartum.


Introduction of glucan synthase into the cytosol in wheat endosperm causes massive maltose accumulation and represses starch synthesis.
We expressed a bacterial glucan synthase (Agrobacterium GlgA) in the cytosol of developing endosperm cells in wheat grains, to discover whether it could generate a glucan from cytosolic ADP-glucose. Transgenic lines had high glucan synthase activity during grain filling, but did not accumulate glucan. Instead, grains accumulated very high concentrations of maltose. They had large volumes during development due to high water content, and very shrivelled grains at maturity. Starch synthesis was severely reduced. We propose that cytosolic glucan synthesized by the glucan synthase was immediately hydrolysed to maltose by cytosolic β-amylase(s). Maltose accumulation resulted in a high osmotic potential in developing grain, drawing in excess water that stretched the seed coat and pericarp. Loss of water during grain maturation then led to shrinkage when the grains matured. Maltose accumulation is likely to account for the reduced starch synthesis in transgenic grains, through signalling and toxic effects. Using bioinformatics, we identify an isoform of β-amylase likely to be responsible for maltose accumulation. Removal of this isoform through identification of TILLING mutants or genome editing, combined with co-expression of heterologous glucan synthase and a glucan branching enzyme, may in future enable elevated yields of carbohydrate through simultaneous accumulation of starch and cytosolic glucan.


Concomitant anal and cervical human papillomavirusV infections and intraepithelial neoplasia in HIV-infected and uninfected women.
To assess factors associated with concomitant anal and cervical human papillomavirus (HPV) infections in HIV-infected and at-risk women. A study nested within the Women's Interagency HIV Study (WIHS), a multicenter longitudinal study of HIV-1 infection in women conducted in six centers within the United States. Four hundred and seventy HIV-infected and 185 HIV-uninfected WIHS participants were interviewed and examined with anal and cervical cytology testing. Exfoliated cervical and anal specimens were assessed for HPV using PCR and type-specific HPV testing. Women with abnormal cytologic results had colposcopy or anoscopy-guided biopsy of visible lesions. Logistic regression analyses were performed and odds ratios (ORs) measured the association for concomitant anal and cervical HPV infection. One hundred and sixty-three (42%) HIV-infected women had detectable anal and cervical HPV infection compared with 12 (8%) of the HIV-uninfected women (P < 0.001). HIV-infected women were more likely to have the same human papillomavirus (HPV) genotype in the anus and cervix than HIV-uninfected women (18 vs. 3%, P < 0.001). This was true for both oncogenic (9 vs. 2%, P = 0.003) and nononcogenic (12 vs. 1%, P < 0.001) HPV types. In multivariable analysis, the strongest factor associated with both oncogenic and nononcogenic concomitant HPV infection was being HIV-infected (OR = 4.6 and OR = 16.9, respectively). In multivariable analysis of HIV-infected women, CD4 cell count of less than 200 was the strongest factor associated with concomitant oncogenic (OR = 4.2) and nononcogenic (OR = 16.5) HPV infection. HIV-infected women, particularly those women with low CD4 cell counts, may be good candidates for HPV screening and monitoring for both cervical and anal disease.


Awareness and opinions regarding contraception by women of reproductive age in North-West Nigeria.
North-west Nigeria is a traditionally high fertility setting. Increasing attention is being channeled into scaling up the various interventions that can reduce high fertility, including enhancing contraceptive uptake among women of reproductive age. However, in order to improve demand for contraception, understanding the level of awareness, knowledge and perception of WRA to contraceptive use is essential. This study examines the level of knowledge and perception of WRA to contraceptive use. A descriptive cross-sectional study was carried out in December 2016 in Kebbi and Sokoto states, North-west Nigeria. Semi-structured questionnaires were administered to 500 women between the ages of 15-49 years in 4 randomly selected local government areas across the states. Data analysis included descriptive and bi-variate statistics. Only 43.8% claimed to be currently using a modern form of contraception; 82.4% were aware of at least one form of contraception, while health personnel formed the major source of information. The most popular and least common modern contraceptive methods were the male condom (86.8%) and vasectomy (26.6%) respectively. A higher proportion of married respondents (88.8%) than those presently single (68.8%) had good knowledge of contraceptives. Perception of contraceptives was mixed, with majority believing that contraceptives should be made available to every woman, despite stating that it contradicted their religious beliefs. These results underline the need for policy makers to focus on improving the knowledge of younger age groups on contraceptives and include other information channels such as peer-to-peer discussions to increase awareness.


IL-22 ameliorates intestinal inflammation in a mouse model of ulcerative colitis.
Expression of IL-22 is induced in several human inflammatory conditions, including inflammatory bowel disease (IBD). Expression of the IL-22 receptor is restricted to innate immune cells; however, the role of IL-22 in colitis has not yet been defined. We developed what we believe to be a novel microinjection-based local gene-delivery system that is capable of targeting the inflamed intestine. Using this approach, we demonstrated a therapeutic potency for IL-22-mediated activation of the innate immune pathway in a mouse model of Th2-mediated colitis that induces disease with characteristics similar to that of IBD ulcerative colitis (UC). IL-22 gene delivery enhanced STAT3 activation specifically within colonic epithelial cells and induced both STAT3-dependent expression of mucus-associated molecules and restitution of mucus-producing goblet cells. Importantly, IL-22 gene delivery led to rapid amelioration of local intestinal inflammation. The amelioration of disease by IL-22 was mediated by enhanced mucus production. In addition, local gene delivery was used to inhibit IL-22 activity through overexpression of IL-22-binding protein. Treatment with IL-22-binding protein suppressed goblet cell restitution during the recovery phase of a dextran sulfate sodium-induced model of acute colitis. These data demonstrate what we believe to be a novel function for IL-22 in the intestine and suggest the potency of a local IL-22 gene-delivery system for treating UC.


Alpha-Toxin Contributes to Biofilm Formation among Staphylococcus aureus Wound Isolates.
Biofilms complicate treatment of Staphylococcus aureus (SA) wound infections. Previously, we determined alpha-toxin (AT)-promoted SA biofilm formation on mucosal tissue. Therefore, we evaluated SA wound isolates for AT production and biofilm formation on epithelium and assessed the role of AT in biofilm formation. Thirty-eight wound isolates were molecularly typed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (ST), and spa typing. We measured biofilm formation of these SA isolates in vitro and ex vivo and quantified ex vivo AT production. We also investigated the effect of an anti-AT monoclonal antibody (MEDI4893*) on ex vivo biofilm formation by methicillin-resistant SA (USA 300 LAC) and tested whether purified AT rescued the biofilm defect of hla mutant SA strains. The predominant PFGE/ST combinations were USA100/ST5 (50%) and USA300/ST8 (33%) for methicillin-resistant SA (MRSA, n = 18), and USA200/ST30 (20%) for methicillin-susceptible SA (MSSA, n = 20). Ex vivo AT production correlated significantly with ex vivo SA wound isolate biofilm formation. Anti-alpha-toxin monoclonal antibody (MEDI4893*) prevented ex vivo biofilm formation by MRSA USA300 strain LAC. Wild-type AT rescued the ex vivo biofilm defect of non-AT producing SA strains. These findings provide evidence that AT plays a role in SA biofilm formation on epithelial surfaces and suggest that neutralization of AT may be useful in preventing and treating SA infections.


Efficacy of prophylactic mastectomy in women with unilateral breast cancer: a cancer research network project.
We investigated the efficacy of contralateral prophylactic mastectomy (CPM) in reducing contralateral breast cancer incidence and breast cancer mortality among women who have already been diagnosed with breast cancer. This retrospective cohort study comprised approximately 50,000 women who were diagnosed with unilateral breast cancer during 1979 to 1999. Using computerized data confirmed by chart review, we identified 1,072 women (1.9%) who had CPM. We obtained covariate information for these women and for a sample of 317 women who did not undergo CPM. The median time from initial breast cancer diagnosis to the end of follow-up was 5.7 years. Contralateral breast cancer developed in 0.5% of women with CPM, metastatic disease developed in 10.5%, and subsequent breast cancer developed in 12.4%; 8.1% died from breast cancer. Contralateral breast cancer developed in 2.7% of women without CPM, and 11.7% died of breast cancer. After adjustment for initial breast cancer characteristics, treatment, and breast cancer risk factors, the hazard ratio (HR) for the occurrence of contralateral breast cancer after CPM was 0.03 (95% CI, 0.006 to 0.13). After adjustment for breast cancer characteristics and treatment, the HRs for the relationship of CPM with death from breast cancer, with death from other causes, and with all-cause mortality were 0.57 (95% CI, 0.45 to 0.72), 0.78 (95% CI, 0.57 to 1.06), and 0.60 (95% CI, 0.50 to 0.72), respectively. CPM seems to protect against the development of contralateral breast cancer, and although women who underwent CPM had relatively low all-cause mortality, CPM also was associated with decreased breast cancer mortality.


Microsporidiosis in zebrafish research facilities.
Pseudoloma neurophilia (Microsporidia) is the most common pathogen detected in zebrafish (Danio rerio) from research facilities. The parasite infects the central nervous system and muscle and may be associated with emaciation and skeletal deformities. However, many fish exhibit subclinical infections. Another microsporidium, Pleistophora hyphessobryconis, has recently been detected in a few zebrafish facilities. Here, we review the methods for diagnosis and detection, modes of transmission, and approaches used to control microsporidia in zebrafish, focusing on P. neurophilia. The parasite can be readily transmitted by feeding spores or infected tissues, and we show that cohabitation with infected fish is also an effective means of transmission. Spores are released from live fish in various manners, including through the urine, feces, and sex products during spawning. Indeed, P. neurophilia infects both the eggs and ovarian tissues, where we found concentrations ranging from 12,000 to 88,000 spores per ovary. Hence, various lines of evidence support the conclusion that maternal transmission is a route of infection: spores are numerous in ovaries and developing follicles in infected females, spores are present in spawned eggs and water from spawning tanks based on polymerase chain reaction tests, and larvae are very susceptible to the infection. Furthermore, egg surface disinfectants presently used in zebrafish laboratories are ineffective against microsporidian spores. At this time, the most effective method for prevention of these parasites is avoidance.


G-protein-independent signaling by G-protein-coupled receptors.
Two classes of receptors transduce neurotransmitter signals: ionotropic receptors and heptahelical metabotropic receptors. Whereas the ionotropic receptors are structurally associated with a membrane channel, a mediating mechanism is necessary to functionally link metabotropic receptors with their respective effectors. According to the accepted paradigm, the first step in the metabotropic transduction process requires the activation of heterotrimeric G-proteins. An increasing number of observations, however, point to a novel mechanism through which neurotransmitters can initiate biochemical signals and modulate neuronal excitability. According to this mechanism metabotropic receptors induce responses by activating transduction systems that do not involve G-proteins.


Reverse genetics screen in zebrafish identifies a role of miR-142a-3p in vascular development and integrity.
MicroRNAs are a well-studied class of non-coding RNA and are known to regulate developmental processes in eukaryotes. Their role in key biological processes such as vasculature development has attracted interest. However, a comprehensive understanding of molecular regulation of angiogenesis and vascular integrity during development remains less explored. Here we identified miRNAs involved in the development and maintenance of vasculature in zebrafish embryos using a reverse genetics approach. Using a combination of bioinformatics predictions and literature based evidences we mined over 701 Human and 329 Zebrafish miRNAs to derive a list of 29 miRNAs targeting vascular specific genes in zebrafish. We shortlisted eight miRNAs and investigated their potential role in regulating vascular development in zebrafish transgenic model. In this screen we identified three miRNAs, namely miR-1, miR-144 and miR-142a-3p that have the potential to influence vascular development in zebrafish. We show that miR-142a-3p mediates vascular integrity and developmental angiogenesis in vivo. Overexpression of miR-142a-3p results in loss of vascular integrity, hemorrhage and vascular remodeling during zebrafish embryonic development, while loss of function of miR-142a-3p causes abnormal vascular remodeling. MiR-142a-3p functions in part by directly repressing cdh5 (VE-cadherin). The vascular abnormalities that results from modulation of miR-142a-3p are reminiscent of cdh5 perturbation in zebrafish embryos. We also demonstrate that the action of miR-142a on cdh5 is potentially regulated by Lmo2, an important transcription factor, known for its role in vasculature development. The miR142a-3p mediated control of cdh5 constitutes an additional layer of regulation for maintaining vascular integrity and developmental angiogenesis. These findings have implications in development, wound repair and tumor growth.


Diversity in the medical office: meeting the needs of the new patients.
The 2000 Census provided information on changing patterns of immigration and internal migration within the United States. As the population shifts to be diverse in more places than ever before, medical offices will need to be ready to meet the needs of a new population of patients. Since more than two-thirds of the counties in our country today have measurable minority populations, even the small community office or clinic will need to be among those making adjustments to reach out to these changing populations.


Migration of leukocytes across endothelium and beyond: molecules involved in the transmigration and fate of monocytes.
Passage of leukocytes across the endothelial lining into sites of inflammation has been shown to be regulated largely by platelet/endothelial cell adhesion molecule-1 (PECAM/CD31). We summarize recent work from our laboratory documenting that PECAM is involved both in diapedesis and the subsequent step of migration across the basal lamina. The former process involves a homophilic interaction between the amino-terminal extracellular domains of PECAM on the leukocyte and on the endothelial cell. The latter process involves a heterophilic interaction between the membrane-proximal extracellular domain of PECAM and an unknown ligand(s) in the subendothelial basal lamina. These findings are demonstrated in both in vitro and in vivo models. For monocytes, however, transmigration is just the first step in the next phase of their lives. In addition to their specific recruitment during the inflammatory response, many monocytes constitutively leave the bloodstream to enter tissues. However, only a fraction of these become tissue macrophages. In an in vitro model of monocyte emigration, approximately half of the leukocytes that initially transmigrate an endothelial monolayer migrate back out in the basal-to-apical direction within the next 2 days. This reverse transmigration cannot be blocked by anti-PECAM reagents, but involves p-glycoprotein and tissue factor expressed on the leukocytes. The reverse transmigrating cells are phenotypically dendritic cells (DC). Their maturation to mature DC can be accelerated by inclusion of inflammatory stimuli in the co-culture. The cells that remain behind in the subendothelial collagen are phenotypically macrophages. We postulate that a major source of DC in lymph nodes is cells derived from monocytes that enter a tissue via the blood and leave several days later via afferent lymphatic channels.


Structure and expression of the Saccharomyces cerevisiae CRY1 gene: a highly conserved ribosomal protein gene.
The Saccharomyces cerevisiae CRY1 gene encodes ribosomal protein rp59, a component of the 40S ribosomal subunit. Mutations in CRY1 can confer resistance to the alkaloid cryptopleurine, an inhibitor of the elongation step of translation. The nucleotide sequence of the cloned CRY1 gene was determined. The predicted amino acid sequence shows that CRY1 encodes a 14,561-dalton polypeptide that has 88% amino acid sequence homology to the hamster or human S14 ribosomal protein responsible for emetine resistance and 45% homology to Escherichia coli ribosomal protein S11. Analysis of the DNA sequences upstream from CRY1 revealed the presence of three sequences, HOMOL1 (consensus, A/TACATCC/TG/ATA/GCA), RPG (consensus, ACCCA/GTACATT/CT/A), and a thymine-rich sequence, found upstream of more than 20 other cloned yeast genes encoding components of the translational apparatus. We exploited the ability to assay the expression of CRY1 in vivo by using the cryptopleurine resistance phenotype to demonstrate that these three consensus sequences are necessary for the transcription of CRY1. We previously showed that the upstream promoter element of the yeast RP39A gene consists of these identical sequence motifs. Therefore, we suggest that these three sequences define a consensus promoter element for the genes encoding the yeast translational apparatus. CRY1 is one of several hundred yeast genes, including ribosomal protein genes, whose expression is transiently decreased 10-fold upon heat shock. We found that the HOMOL1 and RPG consensus sequences are not necessary for the heat shock response of CRY1.


Morphology of elastic nematic liquid crystal membranes.
Liquid crystalline phases found in many biological materials, such as actin, DNA, cellulose, and collagen can be responsible for the deformation of cell membranes. In this paper, cell membrane deformation is investigated through the coupling between liquid crystal anisotropy and membrane bending elasticity. The generalized shape equation for anisotropic interfaces, which resort to the Cahn-Hoffman capillarity vector, the Rapini-Papoular anchoring energy, and the Helfrich elastic energy, is applied to gain insight into the deformation of closed liquid crystal membranes. This study presents a general morphological phase diagram of membrane surface patterns, in which two characteristic regimes of membrane shapes can be classified with respect to the most dominant factor between liquid crystal anisotropy and bending elasticity. To that end, we consider a 2D nematic liquid crystal droplet immersed in a isotropic phase in the presence of an interfacial layer of surfactants, which leads to an additional elastic contribution to the free energy of the system. The presented results indicate that, depending on the bending elasticity of the cell membrane, the liquid crystal might be able to deform the cell, thereby resulting in anisotropic asymmetric shapes. As liquid crystal anisotropy dominates the bending elasticity, spindle-like or tactoid shapes, which are extensively observed in experiments, can be formed. The findings provide a foundational framework to better understand membrane topologies in living soft matters. Furthermore, the coupling between order and curvature of membranes shed new light into the design of novel functional soft materials.


Cost, performance, and reliability simulator for optical transceiver modules.
The program couple combines simulators for optical performance, mechanical reliability, and production cost under a graphical-user interface to design, simulate, and evaluate micro-optomechanical structures. The thermal simulator predicts the package temperature distribution on the basis of the materials and the geometry as well as on heat sources, sinks, and boundary conditions. The thermal distribution is input to the mechanical simulator, which calculates the stresses or strains and displacements caused by differential thermal expansion. The optical simulator predicts the impact on the optical efficiency and the cross talk of mechanical and optical parameter variations such as solder heights, misalignments, and wavelength distributions. The cost simulator represents the manufacturing process flow and calculates the final cost and the cost sensitivity on basis of the cost and the yield of each process step. By means of balancing detector and coupling yield, cosimulation from optical to cost domains determines the optimum detector size to produce the lowest-cost transceiver module.


Elevated levels of somatic mutation in a manifesting BRCA1 mutation carrier.
Homozygous loss of activity at the breast cancerpredisposing genes BRCA1 and BRCA2 (FANCD1) confers increased susceptibility to DNA double strand breaks, but this genotype occurs only in the tumor itself, following loss of heterozygosity at one of these loci. Thus, if these genes play a role in tumor etiology as opposed to tumor progression, they must manifest a heterozygous phenotype at the cellular level. To investigate the potential consequences of somatic heterozygosity for a BRCA1 mutation demonstrably associated with breast carcinogenesis on background somatic mutational burden, we applied the two standard assays of in vivo human somatic mutation to blood samples from a manifesting carrier of the Q1200X mutation in BRCA1 whose tumor was uniquely ascertained through an MRI screening study. The patient had an allele-loss mutation frequency of 19.4 x 10(-6) at the autosomal GPA locus in erythrocytes and 17.1 x 10(-6) at the X-linked HPRT locus in lymphocytes. Both of these mutation frequencies are significantly higher than expected from age-matched disease-free controls (P < 0.05). Mutation at the HPRT locus was similarly elevated in lymphoblastoid cell lines established from three other BRCA1 mutation carriers with breast cancer. Our patient's GPA mutation frequency is below the level established for diagnosis of homozygous Fanconi anemia patients, but consistent with data from obligate heterozygotes. The increased HPRT mutation frequency is more reminiscent of data from patients with xeroderma pigmentosum, a disease characterized by UV sensitivity and deficiency in the nucleotide excision pathway of DNA repair. Therefore, this BRCA1-associated breast cancer patient manifests a unique phenotype of increased background mutagenesis that likely contributed to the development of her disease independent of loss of heterozygosity at the susceptibility locus.


The Stockholm spinal cord injury study: 1. Medical problems in a regional SCI population.
Out of a regional traumatic spinal cord injury population consisting of 379 individuals, 353 (93.1%) participated in the present study. Subjects were individually interviewed using semi-structured protocols. In addition, previous medical records were available for over 96% of subjects, and were used in all these cases to minimise recall bias. Cause of injury, prevalence of present medical symptoms and occurrence of medical complications in the post-acute, post-discharge phase were recorded. Neurological classification was verified by physical examination according to ASIA/IMSOP standards. Many subjects had experienced complications since discharge from initial hospitalisation, especially urinary tract infections, decubitus ulcers, urolithiasis, and neurological deterioration. Prevalence of medical symptoms was also high. More than 41% of subjects with spastic paralysis reported excessive spasticity to be associated with additional functional impairment and/or pain. Almost two-thirds of subjects reported significant pain, with a predominance of neurogenic-type pain. Bladder and bowel dysfunction were each rated by nearly 41% of subjects as a moderate to severe life problem. As expected, sexual dysfunction was also commonly reported. Prevalence of reported symptoms by general systems review was high, particularly fatigue, constipation, ankle oedema, joint and muscle problems, and disturbed sleep. However, lack of adequate normative data precludes comparison with the general population. The frequent occurrence of reported medical problems and complications support advocacy of comprehensive, life-long care for SCI patients. The commonly reported problems of neurogenic pain and neurological deterioration, in particular, require more attention, as these symptoms are not seldom ominous, either by virtue of their impact on quality of life, or because of underlying pathology.


Somatotopic dominance in tactile temporal processing.
The sense of touch is initiated by stimulation of peripheral mechanoreceptors, and then the spatio-temporal pattern of the receptors' activation is interpreted by central cortical processing. To explore the tactile central processing, we psychophysically studied human judgments of the temporal relationships between two tactile events occurring at different skin locations. We examined four types of two-point temporal judgments-simultaneity, temporal order, apparent motion, and inter-stimulus interval-which differ from one another in time scale and task requirement. To perform any of the four temporal judgment tasks, the brain has to integrate spatially separated inputs. The main focus of the present study is to examine how the spatial separation affects the temporal judgment tasks. Two spatial coordinates can be defined in touch: the somatotopic coordinate, defined by cortical topography, and the spatiotopic coordinate, defined in the environment. In our experiments, the somatotopic distance was manipulated by stimulating the middle and index fingers of the same hand or different hands (ipsilateral vs. bilateral conditions), while the spatiotopic distance was manipulated by increasing the stimulators' separation under bilateral conditions (bilateral-near vs. bilateral-far conditions). Our results clearly demonstrated that all four of the temporal judgments were significantly affected by the somatotopic distance, but only slightly by the spatiotopic distance. The present results, together with the previous findings, suggest that tactile temporal judgments in a wide range of time scale, from several to several 100 ms, primarily reflect processing at the level of somatotopic representation unless the performance is further constrained by spatial processing.


Quantitative evaluation of software packages for single-molecule localization microscopy.
The quality of super-resolution images obtained by single-molecule localization microscopy (SMLM) depends largely on the software used to detect and accurately localize point sources. In this work, we focus on the computational aspects of super-resolution microscopy and present a comprehensive evaluation of localization software packages. Our philosophy is to evaluate each package as a whole, thus maintaining the integrity of the software. We prepared synthetic data that represent three-dimensional structures modeled after biological components, taking excitation parameters, noise sources, point-spread functions and pixelation into account. We then asked developers to run their software on our data; most responded favorably, allowing us to present a broad picture of the methods available. We evaluated their results using quantitative and user-interpretable criteria: detection rate, accuracy, quality of image reconstruction, resolution, software usability and computational resources. These metrics reflect the various tradeoffs of SMLM software packages and help users to choose the software that fits their needs.


Long-term study of Cryptosporidium and Giardia occurrence and quantitative microbial risk assessment in surface waters of Arizona in the USA.
A four-year systematic study of microbial water quality in the surface waters of central Arizona was performed. The objectives of this study were to investigate the occurrence of microbial pathogens and indicators in the waters. A total of 192 water samples from five sites were analyzed for Cryptosporidium, Giardia, and various microbial and physical indicators. Ten percent (16/162) of the samples collected using EnviroChek filters were positive for Cryptosporidium oocysts, whereas no oocysts (<1 in 5 L) were detected in the grab samples (0/30). Giardia cysts were detected in 10% (3/30) of the grab samples and in 27% (44/162) samples collected using EnviroChek filters. Mean concentrations of oocysts in the source waters at the treatment plants were lower than the Bin 1 category of the USEPA Long Term 2 Enhanced Surface Water Treatment Rule; therefore no additional treatment is required by the plants. The annual risks of infection from Cryptosporidium met the annual acceptable risk of 10(-4) at all sampling sites, whereas the risks of Giardia infection at the Verde River and the Salt River were 5.70E-04 and 2.66E-04, respectively.


How does information influence hope in family members of traumatic coma patients in intensive care unit?
To assess the interplay between hope and the information provided by health care professionals. Earlier research learned that hope is crucial for relatives of traumatic coma patients. Also it has been reported that the need for information is extremely important for relatives of critically ill patients. A qualitative approach according to the 'grounded theory' method with constant comparison was used. We held 24 in-depth interviews with 22 family members of 16 patients with traumatic coma. Data processing and data analysis took place in a cyclic process wherein the induction of themes was alternated by confrontation with new material. Family members of traumatic coma patients want information that is as accurate as possible, provided by doctors and nurses in an understandable manner and leaving room for hope. At first, family members can do no more than passively absorb the information they receive. After some time, they actively start working with information and learn what to build their hope on. In this way, concrete hope evolves and seems to be strongly determined by information. Information that is more positive than warranted is not appreciated at all. It leads to false hope and once its real nature becomes apparent, to increased distress and loss of trust in the professionals. The process of hope is crucial in coping with traumatic coma and information can facilitate this process. If professionals, especially nurses, keep the process in mind that family members go through in handling information, they can not only facilitate this process but also help them to establish realistic hope.


Sexual dimorphism in Th17/Treg axis in lymph nodes draining inflamed joints in rats with collagen-induced arthritis.
Collagen type II-induced arthritis (CIA) in Dark Agouti rats, a model of rheumatoid arthritis (RA), reproduces sexual dimorphism in the incidence and severity of the human disease. Th17 cells are central in the induction/propagation of autoimmune inflammation in CIA and RA. To assess mechanisms underlying this dimorphism in CIA rats, in lymph nodes draining inflamed joints and adjacent tissues (dLNs) from CIA rats of both sexes Th17/CD25+Foxp3+CD4+ T-regulatory cell (Treg) ratio, Th17 cell redifferentiation in functionally distinct subsets and Treg transdifferentiation into IL-17-producing cells (exTregs) were examined. In female rats (developing more severe CIA than their male counterparts) the higher frequency of all Th17 cells (reflecting partly their greater proliferation), followed by the higher frequency of highly pathogenic IFN-γ/GM-CSF-co-producing cells, but lower frequency of less pathogenic/immunoregulatory IL-10-producing cells among them was found. Additionally, compared with male rats, in female rats the lower frequency of Tregs was observed. Moreover, Tregs from female rats exhibited diminished proliferative and suppressive capacity (judging by PD-1 expression) and enhanced conversion into IL-17-producing cells. Given that TGF-β concentration was comparable in collagen-type II-stimulated dLN cell cultures from female and male rats, the shift in Th17/Treg ratio followed by augmented Th17 cell redifferentiation into IFN-γ/GM-CSF-co-producing cells and Treg transdifferentiation into IL-17-producing cells in female rats was associated with increased concentration of IL-6 in female rat dLN cell cultures, and the higher frequency of IL-1β- and IL-23-producing cells among their dLN cells. The lower frequency of IL-10-producing B cells, presumably B regulatory cells (Bregs) could also contribute to the shift in Th17/Treg ratio in female rat compared with male rat dLNs. Consistently, the lower expression of IL-35 (the cytokine promoting Treg expansion directly and indirectly, by favoring Breg expansion and conversion into IL-10/IL-35-producing cells) in female rat dLN cells was detected. Thus, the study identified putative cellular and molecular substrates of the sexual dimorphism in the immunopathogenesis and clinical outcome of CIA and suggested mechanisms to be targeted in females to improve control of Th17 response, and consequently clinical outcome of CIA, and possibly RA.


Glucocorticoid ultradian rhythmicity differentially regulates mood and resting state networks in the human brain: A randomised controlled clinical trial.
Adrenal glucocorticoid secretion into the systematic circulation is characterised by a complex rhythm, composed of the diurnal variation, formed by changes in pulse amplitude of an underlying ultradian rhythm of short duration hormonal pulses. To elucidate the potential neurobiological significance of glucocorticoid pulsatility in man, we have conducted a randomised, double-blind, placebo-controlled, three-way crossover clinical trial on 15 healthy volunteers, investigating the impact of different glucocorticoid rhythms on measures of mood and neural activity under resting conditions by recruiting functional neuroimaging, computerised behavioural tests and ecological momentary assessments. Endogenous glucocorticoid biosynthesis was pharmacologically suppressed, and plasma levels of corticosteroid restored by hydrocortisone replacement in three different regimes, either mimicking the normal ultradian and circadian profile of the hormone, or retaining the normal circadian but abolishing the ultradian rhythm of the hormone, or by our current best oral replacement regime which results in a suboptimal circadian and ultradian rhythm. Our results indicate that changes in the temporal mode of glucocorticoid replacement impact (i) the morning levels of self-perceived vigour, fatigue and concentration, (ii) the diurnal pattern of mood variation, (iii) the within-network functional connectivity of various large-scale resting state networks of the human brain, (iv) the functional connectivity of the default-mode, salience and executive control networks with glucocorticoid-sensitive nodes of the corticolimbic system, and (v) the functional relationship between mood variation and underlying neural networks. The findings indicate that the pattern of the ultradian glucocorticoid rhythm could affect cognitive psychophysiology under non-stressful conditions and opens new pathways for our understanding on the neuropsychological effects of cortisol pulsatility with relevance to the goal of optimising glucocorticoid replacement strategies.


Administration of drug and nutritional components in nano-engineered form to increase delivery ratio and reduce current inefficient practice.
The article critically discusses parenteral delivery of self-assembled lipid or amphiphile nanoparticles, in the form of aggregated clusters or particles (capsules). The end-product or drug form is for application by administration of the medicine active in encapsulated form. This is used for site-specific cell manipulation and clinical therapy and introduces this directly to the body via the systemic route. The technology discussed represents a platform formulation that can be modified for a range of specific cellular targets. The components of the nanoparticle are assembled piece-by-piece and this provides an element of design flexibility, with the core particle being built-up in a succession of layers to ensure circulatory longevity and storage stability. This strategy excludes a more generalised delivery and widespread lack of active targeting and thus low dosage rather than avoidance of target, which is at best detrimental and at worst catastrophic in terms of non-targeted cell death. However, in some cases such as the AmBisome nanoparticle this drug delivery approach can work. This "better focussing" is achieved by a dual use of i) biocompatible particle coating chemistry and a ii) cell-ligand imprinted nanoparticle surface that furnishes the engineered nanoparticle with a recognition element to form a complex but more efficacious dispersible fusogenic pro-drug moiety. Non-targeted delivery of drugs such as those commonly forming the basis of transdermal delivery have been generically based on topical or adhesive patch-based delivery (emulsions) systems. This procedure even with recent advancements and patents is customarily inefficient in dosage and payload delivery, inconsistent in terms of product potency and inflexible to further modification or purpose-related enhancement. The assembly and delivery methodologies discussed here take the new experimental medicine and review them in a more focused and purposeful therapy-constructed manner. It is the use of a smart platform technology that is responsible for increased efficacy when applied to the disorder of interest.


Family of a person suffering from schizophrenia in the context of a qualitative approach to understanding the family as a system.
Qualitative analysis of the family system including a person suffering from schizophrenia is an underestimated research paradigm in modern psychiatry, clinical psychology and psychotherapy. This method is important both from the cognitive point of view and for evaluation of the effectiveness of therapy. The aim of the study was a qualitative analysis of the therapeutic process in a family whose member is suffering from schizophrenic psychosis. The study of the process using qualitative research methodology in the paradigm of systemic thinking pointing to the mutual interactions in the family system inducing relapses in the identified patient, with identification of triangulation processes, transgenerational myths inheritance and coalitions formed by the family members. The course of a therapeutic process of the family consisting of a parental couple and an adult daughter who was a patient with schizophrenia was investigated. Relapses of the daughter's disease occurred at the time of over-controlling attitude of the mother. Excessive control was associated with the daughter's immediate cut off from the family, discontinuation of medication, and a relapse. The relapse meant a repeated attempt of the mother to take control over the daughter's medication. The analyzed family is genetically predisposed to the development of schizophrenic psychosis. Schizophrenia in the daughter is likely to have a neurodevelopmental background - it has developed as a result of the interaction of biological factors (genetic predisposition) and factors that interfered with the intrauterine development of the fetus. The burden of mental disorder is inherited transgenerationally in the analyzed family. Excessive anxiety of the mother, exaggerated emotional expression, and attempts to control the treatment lead to relapses of the daughter's disease. Recurrences are a circular process induced by the mother's attitude.


Drag reduction mechanism of Paramisgurnus dabryanus loach with self-lubricating and flexible micro-morphology.
Underwater machinery withstands great resistance in the water, which can result in consumption of a large amount of power. Inspired by the character that loach could move quickly in mud, the drag reduction mechanism of Paramisgurnus dabryanus loach is discussed in this paper. Subjected to the compression and scraping of water and sediments, a loach could not only secrete a lubricating mucus film, but also importantly, retain its mucus well from losing rapidly through its surface micro structure. In addition, it has been found that flexible deformations can maximize the drag reduction rate. This self-adaptation characteristic can keep the drag reduction rate always at high level in wider range of speeds. Therefore, even though the part of surface of underwater machinery cannot secrete mucus, it should be designed by imitating the bionic micro-morphology to absorb and store fluid, and eventually form a self-lubrication film to reduce the resistance. In the present study, the Paramisgurnus dabryanus loach is taken as the bionic prototype to learn how to avoid or slow down the mucus loss through its body surface. This combination of the flexible and micro morphology method provides a potential reference for drag reduction of underwater machinery.


Interactive effects of hypoxia and polybrominated diphenyl ethers (PBDEs) on microbial community assembly in surface marine sediments.
Hypoxia alters the oxidation-reduction balance and the biogeochemical processes in sediments, but little is known about its impacts on the microbial community that is responsible for such processes. In this study, we investigated the effects of hypoxia and the ubiquitously dispersed flame-retardant BDE47 on the bacterial communities in marine surface sediments during a 28-days microcosm experiment. Both hypoxia and BDE47 alone significantly altered the bacterial community and reduced the species and genetic diversity. UniFrac analysis revealed that BDE47 selected certain bacterial species and resulted in major community shifts, whereas hypoxia changed the relative abundances of taxa, suggesting slower but nonetheless significant community shifts. These two stressors targeted mostly different taxa, but they both favored Bacteroidetes and suppressed Gammaproteobacteria. Importantly, the impacts of BDE47 on bacterial communities were different under hypoxic and normoxic conditions, highlighting the need to consider risk assessments for BDE47 in a broader context of interaction with hypoxia.


Quantitative Analysis of the Microtubule Interaction of Rabies Virus P3 Protein: Roles in Immune Evasion and Pathogenesis.
Although microtubules (MTs) are known to have important roles in intracellular transport of many viruses, a number of reports suggest that specific viral MT-associated proteins (MAPs) target MTs to subvert distinct MT-dependent cellular processes. The precise functional importance of these interactions and their roles in pathogenesis, however, remain largely unresolved. To assess the association with disease of the rabies virus (RABV) MAP, P3, we quantitatively compared the phenotypes of P3 from a pathogenic RABV strain, Nishigahara (Ni) and a non-pathogenic Ni-derivative strain, Ni-CE. Using confocal/live-cell imaging and dSTORM super-resolution microscopy to quantify protein interactions with the MT network and with individual MT filaments, we found that the interaction by Ni-CE-P3 is significantly impaired compared with Ni-P3. This correlated with an impaired capacity to effect association of the transcription factor STAT1 with MTs and to antagonize interferon (IFN)/STAT1-dependent antiviral signaling. Importantly, we identified a single mutation in Ni-CE-P3 that is sufficient to inhibit MT-association and IFN-antagonist function of Ni-P3, and showed that this mutation alone attenuates the pathogenicity of RABV. These data provide evidence that the viral protein-MT interface has important roles in pathogenesis, suggesting that this interface could provide targets for vaccine/antiviral drug development.


Activation of protease-activated receptors by gingipains from Porphyromonas gingivalis leads to platelet aggregation: a new trait in microbial pathogenicity.
The bacterium Porphyromonas gingivalis is a major etiologic agent in the pathogenesis of adult periodontitis in humans. Cysteine proteinases produced by this pathogen, termed gingipains, are considered to be important virulence factors. Among many other potentially deleterious activities, arginine-specific gingipains-R (RgpB and HRgpA) efficiently activate coagulation factors. To further expand knowledge of the interaction between gingipains and the clotting cascade, this study examined their effects on cellular components of the coagulation system. The enzymes induced an increase in intracellular calcium in human platelets at nanomolar concentrations and caused platelet aggregation with efficiency comparable to thrombin. Both effects were dependent on the proteolytic activity of the enzymes. Based on desensitization studies carried out with thrombin and peptide receptor agonists, and immunoinhibition experiments, gingipains-R appeared to be activating the protease-activated receptors, (PAR)-1 and -4, expressed on the surface of platelets. This was confirmed by the finding that HRgpA and RgpB potently activated PAR-1 and PAR-4 in transfected cells stably expressing these receptors. Cumulatively, the results indicate the existence of a novel pathway of host cell activation by bacterial proteinases through PAR cleavage. This mechanism not only represents a new trait in bacterial pathogenicity, but may also explain an emerging link between periodontitis and cardiovascular disease. (Blood. 2001;97:3790-3797)


Dog ecology and demography information to support the planning of rabies control in Machakos District, Kenya.
A study of 150 dog-owning households from six randomly selected sublocations was conducted in Machakos District, Kenya. Initially, all households were visited to collect information on dog ecology and demography based on WHO guidelines and to collect serum for rabies antibody detection. A second visit was made 1 year later, to obtain follow-up data on births, deaths, dog movements and other events since the first visit. Dog ownership was common, with a range of 53--81% (mean=63%) of households owning dogs in the six sublocations. Dog density for the five more rural sublocations ranged from 6 to 21 dogs km(-2) and for the peri-urban sublocation was 110 dogs km(-2). The dog population was estimated to be growing at 9% p.a. (95% C.I. 4--14%). This growth was a function of very high fecundity (1.3 females per female per year) more than compensating for high mortality, particularly among females. Life expectancy from birth was 3.5 years for males and 2.4 years for females. Half the dogs at any one time were less than 1 year of age. All dogs, by design of the study, were owned. Of these, 69% were never restricted and roamed freely to forage for food and mix with other dogs. Only a small proportion of dogs (5%) were fed commercial dog food. Most households reported observing dogs scavenging their garbage, including: their own dogs (81%), their neighbours' dogs (75%) and unknown dogs (45%). Only 29% of dogs at least 3 months of age were reported to be vaccinated against rabies. The proportion vaccinated varied widely between sublocations (5--68%); 48% of dogs reportedly vaccinated had detectable antibodies, 31% at or above levels considered to indicate seroconversion. The proportion of dogs with detectable antibodies declined according to the time since last vaccination (55% if vaccinated < or = 1 year, 47% < or = 2 years and 36% > 2 years); 20% of dogs reported not to have been vaccinated had detectable rabies antibody. Compared to other dog populations in rural eastern and southern Africa, Machakos District has a high density of dogs. The Machakos dog population is growing, highly dynamic, poorly supervised and inadequately vaccinated against rabies. The main implication for rabies control is that adequate vaccination coverage is unlikely to be achieved, even under optimal delivery, using the current strategy of annual vaccination of dogs older than 3 months.


Soluble interleukin-18 receptor complex is a novel biomarker in rheumatoid arthritis.
There has been no report in the literature of a soluble form of interleukin (IL)-18 receptor α (IL-18Rα). In this study, we evaluated the levels and characteristics of soluble IL-18Rα (sIL-18Rα) in the sera of patients with rheumatoid arthritis (RA) and compared these results to control populations. The sIL-18Rα complex was isolated from pooled human blood serum using an anti-IL-18Rα monoclonal antibody affinity column. The purified sIL-18Rα was then examined using Western blot analysis and used in experiments to evaluate the effects on an IL-18-responsive natural killer (NK) human cell line, NK0. An enzyme-linked immunosorbent assay was developed, and sera from 145 patients with RA, 6 patients with adult-onset Still's disease, 31 patients with osteoarthritis (OA), 39 patients with systemic lupus erythematosus (SLE) and 67 controls were tested, along with levels of immunoglobulin M, rheumatoid factor, anticyclic citrullinated peptide antibody, IL-18, IL-13 and interferon (IFN)-γ. Area under the receiver operating characteristic curve (ROC-AUC) analysis was used to evaluate the diagnostic utility of the sIL-18Rα complex. The isolated sIL-18Rα complex can be associated with IL-18 and the soluble form of the IL-18Rβ chain. The sIL-18Rα complex bound to the surface to the NK0 cell line, antagonized the stimulatory effects of IL-18 and IL-2 on the NK0 cell line and inhibited IFN-γ production by the cells. The serum levels of sIL-18Rα complex in RA (186.0 ± 33.5 ng/mL, n = 145) and adult-onset Still's disease (98.2 ± 8.9 ng/mL, n = 6) were significantly (P < 0.001) higher than those in the healthy controls (52.3 ± 8.5 ng/mL, n = 67), OA (38.6 ± 5.4 ng/mL, n = 31), SLE (44.6 ± 3.2 ng/mL, n = 39). The serum level of sIL-18Rα complex was not significantly different between RA and adult-onset Still's disease patients. The serum levels of IL-18, IL-13 and IFN-γ in the RA patients were significantly (P < 0.01) higher than in OA and SLE patients as well as healthy controls. ROC-AUC analysis of the serum concentration of sIL-18Rα indicated that it was significantly diagnostic of RA. Moreover, a tumor necrosis factor inhibitor, etanercept, significantly (P < 0.0001) decreased levels of sIL-18Rα in the sera of 29 RA patients 6 months after treatment. The sIL-18Rα complex could be a potentially useful biomarker for the diagnosis of RA.


Characterization of gene expression profiles in HBV-related liver fibrosis patients and identification of ITGBL1 as a key regulator of fibrogenesis.
Although hepatitis B virus (HBV) infection is the leading cause of liver fibrosis (LF), the mechanisms underlying liver fibrotic progression remain unclear. Here, we investigated the gene expression profiles of HBV-related LF patients. Whole genome expression arrays were used to detect gene expression in liver biopsy samples from chronically HBV infected patients. Through integrative data analysis, we identified several pathways and key genes involved in the initiation and exacerbation of liver fibrosis. Weight gene co-expression analysis revealed that integrin subunit β-like 1 (ITGBL1) was a key regulator of fibrogenesis. Functional experiments demonstrated that ITGBL1 was an upstream regulator of LF via interactions with transforming growth factor β1. In summary, we investigated the gene expression profiles of HBV-related LF patients and identified a key regulator ITGBL1. Our findings provide a foundation for future studies of gene functions and promote the development of novel antifibrotic therapies.


A genome-wide association study identifies five loci influencing facial morphology in Europeans.
Inter-individual variation in facial shape is one of the most noticeable phenotypes in humans, and it is clearly under genetic regulation; however, almost nothing is known about the genetic basis of normal human facial morphology. We therefore conducted a genome-wide association study for facial shape phenotypes in multiple discovery and replication cohorts, considering almost ten thousand individuals of European descent from several countries. Phenotyping of facial shape features was based on landmark data obtained from three-dimensional head magnetic resonance images (MRIs) and two-dimensional portrait images. We identified five independent genetic loci associated with different facial phenotypes, suggesting the involvement of five candidate genes--PRDM16, PAX3, TP63, C5orf50, and COL17A1--in the determination of the human face. Three of them have been implicated previously in vertebrate craniofacial development and disease, and the remaining two genes potentially represent novel players in the molecular networks governing facial development. Our finding at PAX3 influencing the position of the nasion replicates a recent GWAS of facial features. In addition to the reported GWA findings, we established links between common DNA variants previously associated with NSCL/P at 2p21, 8q24, 13q31, and 17q22 and normal facial-shape variations based on a candidate gene approach. Overall our study implies that DNA variants in genes essential for craniofacial development contribute with relatively small effect size to the spectrum of normal variation in human facial morphology. This observation has important consequences for future studies aiming to identify more genes involved in the human facial morphology, as well as for potential applications of DNA prediction of facial shape such as in future forensic applications.


Compliance with an outpatient stress reduction program: rates and predictors of program completion.
The rate at which medical patients physician-referred to an 8-week stress reduction program completed the prescribed intervention was measured and predictors of compliance sought. Seven hundred eighty-four consecutive patients who enrolled in the program over a 2-year period were studied. Of these, 598 (76%) completed the program and 186 (24%) did not. Multiple regression analysis showed that (1) among chronic pain patients, only sex discriminated between completers and noncompleters, with females more than twice as likely to complete the program as males (odds ratio = 2.4; 95% CI = 1.2, 4.4); (2) among patients with stress-related disorders, only the OC scores of the SCL-90-R discriminated between completers and noncompleters (odds ratio = 2.0; 95% CI = 1.2, 3.4). Completion rates for specific diagnoses are reported and discussed. The high rate of completion observed for this intensive program in health behavior change is discussed in terms of the design features and therapeutic modalities of the intervention.


Effects of long-term oral administration of methimazole on femur and tibia properties in male Wistar rats.
Physiological concentrations of thyroid hormones are crucial for skeletal growth and development, physiological bone turnover and bone homeostasis maintenance. Methimazole (1-methyl-2-mercaptoimidazole) is an antithyroid drug used for the treatment of the hyperthyroidism in humans and animals. The aim of the study was to determine effects of long-term oral methimazole treatment in male Wistar rats on biochemical bone metabolism markers, as well as morphological, geometric, densitometric and mechanical properties of femur and tibia. Experimental rats were subjected to 90-day-long oral treatment with 0.05% water solution of methimazole and were kept under identical environmental conditions and received the same diet ad libitum as the control group. Serum concentration of osteocalcin (OC) and C-terminal telopeptides of type I collagen (CTX-I) was determined. Femur and tibia were evaluated using quantitative computed tomography (QCT), peripheral QCT (pQCT) and three-point bending test. Final body weight of the experimental group was significantly decreased by 30% (P=0.01). Methimazole treatment significantly decreased serum OC concentration by 21% (P=0.02) and increased CTX-I concentration by 17% (P=0.06). Methimazole decreased morphological, geometric and densitometric parameters of femur and tibia in rats. Mechanical evaluation of bones has shown significantly decreased maximum elastic strength and ultimate strength of femur in rats treated with methimazole by 36% and 40% when compared to the control group (P<0.05). In conclusion, this study has shown that long-term treatment with methimazole inhibits bone formation and accelerates bone resorption processes. The observed negative effects of methimazole treatment on body weight gain and skeletal properties may be considered as additional possible side effects in living organisms to those reported in the previous studies. It may be suggested that long-term antithyroid treatment should be combined with prevention of the negative effects of methimazole on bone tissue and whole body metabolism.


Stem cell medicine: umbilical cord blood and its stem cell potential.
The ultimate aim of stem cell research is to improve patient outcomes and quality of life, and/or to effect a cure for a variety of inherited or acquired diseases. Improved treatments rely on developments in stem cell therapies and the discovery of new therapeutic drugs that regulate stem cell functions. These complement each other for the repair, regeneration and replacement of damaged or defective tissues. Stem cells may be sourced or derived from blood and tissues postnatally ('adult' stem cells), from the fetus (fetal stem cells) or from the blastocyst in the developing embryo prior to implantation (embryonic stem cells), each forming a unique component of the revolution in stem cell research and therapies. This review will concentrate on recent developments in the use of haemopoietic stem cells from umbilical cord blood for the transplantation of patients with haematological disorders. It will conclude with a summary of the potential of other umbilical cord blood-derived stem cells for tissue repair or regeneration.