Development of GelMA/PCL and dECM/PCL resins for 3D printing of acellular in vitro tissue scaffolds by stereolithography.
Gelatin methacryloyl (GelMA) is a chemically modified extracellular matrix (ECM)-derived biopolymer that is widely used for 3D fabrication of tissue engineering scaffolds. However, its tendency for physical gelation limits its use in aqueous 3D printing resins to low concentrations, yielding a poor printing resolution in stereolithography (SLA). To obtain a GelMA-based resin that can be printed into high-resolution tissue scaffolds, we formulated resins of fish and porcine-derived GelMA in formamide using GelMA alone or mixed with star-shaped poly(ε-caprolactone) methacrylate (PCL-MA). We identified GelMA resins and GelMA/PCL-MA hybrid resins with a ratio of 70/30 wt-% to yield a suitable viscosity for SLA at 32 °C and demonstrated the resolution of the new resins in SLA by 3D printing acellular human small intestine-mimicking tissue scaffolds. The presence of PCL-MA in the hybrid resins improved the 3D printing fidelity compared to the neat GelMA resins, while GelMA provided the hybrid materials with enhanced swelling and proliferation of seeded cells. We further demonstrated the transferability of our resin formulation to native organ-derived materials by successfully replacing GelMA in the hybrid resin with solubilized, methacryloyl-functionalized decellularized liver ECM (dECM-MA) and by 3D printing multi-layer dECM/PCL-MA hydrogels.


Hispanic access to health/mental health services.
Currently, the Hispanic population of the United States is growing very rapidly. Despite the significance of this growth and the fact that it is expected that Hispanics will be soon the largest ethnic minority group in this country, the access to health/mental health care for the Hispanic population is rather limited. Many factors are currently affecting the Hispanics' access to health/mental health care services. Among them, cultural and language barriers, insufficient numbers of Hispanic manpower in the health care professions, low educational and socioeconomic levels, the high number of uninsured Hispanics, and ethnic and racial prejudices and discrimination. In this commentary, I address the factors that interfere with the Hispanics' access to health/mental health care, and advance recommendations geared to alleviate and/or resolve this critical problem.


Ultrastructural analysis of a putative magnetoreceptor in the beak of homing pigeons.
With the use of different light and electron microscopic methods, we investigated the subcellular organization of afferent trigeminal terminals in the upper beak of the homing pigeon, Columba livia, which are about 5 microm in diameter and contain superparamagnetic magnetite (SPM) crystals. The SPM nanocrystals are assembled in clusters (diameter, approximately 1-2 microm). About 10 to 15 of these clusters occur inside one nerve terminal, arranged along the cell membrane. Each SPM cluster is embedded in a solid fibrous cup, open towards the cell surface, to which the cluster adheres by delicate fiber strands. In addition to the SPM clusters, a second inorganic iron compound has been identified: noncrystalline platelets of iron phosphate (about 500 nm wide and long and maximally 100 nm thick) that occur along a fibrous core of the terminal. The anatomic features suggested that these nerve endings could detect small intensity changes of the geomagnetic field. Such stimuli can induce deformations of the SPM clusters, which could be transduced into primary receptor potentials by mechanosensitive membrane receptor channels. The subepidermal fat cells surrounding the nerve endings prevent the inside from external mechanical stimuli. These structural findings corresponded to conclusions inferred from rock magnetic measurements, theoretical calculations, model experiments, and behavioral data, which also matched previous electrophysiologic recordings from migratory birds.


Diaporthe ambigua Associated with Post-Harvest Fruit Rot of Kiwifruit in Chile.
Kiwifruit is an important, expanding commercial crop in Chile. Several fungi have been reported to be associated with post-harvest rots of kiwifruit worldwide (2). During 2011 and 2012, kiwifruit produced in the VI and VII regions of Chile, showing symptoms of inner rot, were investigated with the aim of identifying the disease agent. The affected fruits had brown pubescent skin at the stem end that became soft and lighter in color than the adjacent firm healthy tissues. A watery exudate and white to pale grayish mycelial mats frequently developed at the stem end of the fruit, causing a water-drop stain down the sides on the dry brown healthy skin. The underlying tissue accessed by peeling off the skin was usually water soaked, soft, and lighter green than the healthy tissue. A fermented sour odor occurred frequently on severely decayed fruits. After incubation at 25°C over 7 days on potato dextrose agar (PDA), white to grayish, pale aerial mycelial mats were recovered from fragments of symptomatic kiwifruit superficially disinfected with 95% ethanol. After 2 weeks, black, spherical pycnidia developed, bearing hyaline, ellipsoidal, biguttulate conidia (5.4 to 12.6 × 2.1 to 4.7 μm). After 3 weeks, abundant perithecia embedded in a distinct, black, elevated stroma developed. Perithecia were black, globose, 200 to 500 μm in diameter with necks sinuous, filiform, 550 to 840 × 50 to 95 μm. Clavate, sessile asci, 30.9 to 50.2 × 6.6 to 12.5 μm contained ascospores biseriate, hyaline, smooth, fusoid-ellipsoid, widest just above the septum, tapering towards both ends, medianly septate, constricted at the septum at maturity, with 1 to 2 guttules per cell ascospores, 5.9 to 9.7 × 2.5 to 4.3 μm. All colonies obtained from kiwifruit displayed the same morphological traits and were consistent with those of a Diaporthe sp. (1). The internal transcribed spacer (ITS) region was sequenced using ITS1/ITS4 primers (4). Analysis of ITS region of kiwifruit isolates Damb_12 and Damb_16 (GenBank Accession Nos. KC294545 and KC294544, respectively) revealed 100% nucleotide identity to Diaporthe ambigua (HM575420, HM575419, DQ286274, DQ286270, and DQ286271). Six millimeter plugs from fungal colonies growing on PDA at 25°C for 7 days were used to inoculate 15 healthy untreated, ripe 'Hayward' kiwifruits. Control fruits were inoculated with agar plugs. Inoculated fruits were incubated at 25°C and 80% relative humidity. After 7 days, white to pale grayish mycelial mats developed only on the inoculated fruits, releasing a water drop stain. The underlying tissue was lighter green and water soaked. D. ambigua was reisolated only from the inoculated fruits. D. actinidiae has been previously reported on kiwifruit canes in Chile (3); however, to our knowledge, this is the first report on the occurrence of D. ambigua (Phomopsis ambigua) on kiwifruit in Chile. The fungal isolates (no. 1 to 21) have been deposited in the Laboratorio de Fitopatología Frutal y Molecular, Departamento de Sanidad Vegetal, Facultad de Ciencias Agronómicas de la Universidad de Chile. References: (1) J. C. Jansen van Rensburg et al. Studies in Mycol. 55:65, 2006. (2) L. Luongo et al. J. Plant Pathol. 93:205, 2011. (3) A. Palma and E. Piontelli. E. Bol. Micol. 15:79, 2000. (4) White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.


The Use of Magnetic Resonance Imaging in Axial Spondyloarthritis: Time to Bridge the Gap Between Radiologists and Rheumatologists.
Magnetic resonance imaging (MRI) is involved in the assessment of axial spondyloarthritis (axSpA); however, anecdotal evidence suggests diverse practice among radiologists. The objective of this study was to describe current practice in the use of MRI for assessment of axSpA by UK radiologists. Six hundred ninety-nine UK radiologists were invited to complete an online survey. Availability of MR scanners, familiarity with axSpA disease-specific lesions, and MRI protocols and definitions of positive sacroiliac joint (SIJ) or spinal MRI were assessed. Two-hundred sixty-nine radiologists (38%) from 131/180 (73%) acute UK National Health Service trusts/health boards responded. MRI waiting times < 2 months were reported by 90% of radiologists. Twenty-nine radiologists (11%) used contrast as standard, 256 (91%) used T1 and short-tau inversion recovery, and 172 (64%) also used T2 sequences. Five percent scanned only SIJ, 33% scanned SIJ and lumbar spine, 29% scanned SIJ and thoracolumbar spine, and 30% scanned SIJ and the whole spine. Mean scan time was 34 min. Eighteen percent did not use the subchondral bone marrow edema of the SIJ to help diagnose axSpA and 18% did not use the inflammatory vertebral corner lesions to assist diagnosis. Awareness of axSpA was reported by 75% of radiologists, and awareness of definitions for positive MRI of SIJ and spine by 31% and 25%, respectively. These data highlight the need for better rheumatology-radiology collaboration on the identification of diagnostic axSpA MRI lesions and support the need for a consensus on the most appropriate MRI protocols for the assessment of axSpA.


Bronchodilator Response Assessed by the Forced Oscillation Technique Identifies Poor Asthma Control With Greater Sensitivity Than Spirometry.
Persistent bronchodilator response (BDR) following diagnosis of asthma is an underrecognized treatable trait, associated with worse lung function and asthma control. The forced oscillation technique (FOT) measures respiratory system impedance, and BDR cutoffs have been proposed for healthy adults; however, the relevance in asthma is unknown. We compared BDR cutoffs, using FOT and spirometry, in asthma and the relationship with asthma control. Data from patients with asthma who withheld bronchodilator medication for at least 8 h before a tertiary airway clinic visit were reviewed. All subjects performed FOT and spirometry before and after salbutamol administration, and completed the Asthma Control Test. FOT parameters examined included respiratory system resistance (R5) and reactance (X5) at 5 Hz, and area under the reactance curve (AX). BDR was defined by standard recommendations for spirometry and based on the 95th percentile of BDR in healthy adults for FOT. Fifty-two subjects (18 men; mean age, 53 ± 18 years) were included. BDR was identified more frequently by FOT than spirometry (54% vs 27% of subjects). BDR assessed by X5 and AX, but not R5, was associated with spirometric BDR (χ2, P < .01) and correlated with asthma control (X5: rs = -0.36, P < .01; AX: rs = 0.34, P = .01). BDR measured by reactance parameters identified more subjects with poor asthma control than did spirometry (AX, 69% vs spirometry, 41%). BDR assessed by FOT can identify poor asthma control. Reactance parameters were more sensitive in identifying poor asthma control than spirometry, supporting the use of FOT to complement spirometry in the clinical management of asthma.


Spatial-soliton-induced guided waves in a homogeneous nonlinear Kerr medium.
It has been predicted that an intense pump beam can induce the focusing of a weak probe beam at a different wavelength that is simultaneously propagating in a Kerr homogeneous material through cross-phase modulation. In Kerr-type nonlinear material, spatial solitons are the only stable waves able to propagate at high-intensity levels without leading to catastrophic breakdown. We propose the use of a soliton beam to induce the stable guiding of a probe beam. Experimental results obtained with picosecond pulses are reported that evidence spatial-solitoninduced guiding.


Self-defining memories after severe traumatic brain injury: A preliminary study.
Autobiographical memory plays a major role in the (re)construction of sense of identity, a recurrent issue after a traumatic brain injury (TBI). Although the recall of specific autobiographical events is frequently impaired in patients with TBI, little is known about how these changes affect their sense of self and identity. Thus, we examined self-defining memories (SDMs), that is, the most significant personal memories supporting one's sense of identity, in 16 patients with severe TBI and in matched controls. To this end, participants recalled three SDMs and rated their emotions in response to memory retrieval. In addition, characteristics of SDMs such as specificity, meaning-making, self-connections that reflect identity stability or identity change, content (theme, presence of tension, and redemption sequences) were analysed by independent raters. The main results showed that patients' SDMs were less specific and contained fewer redemption sequences than did those of controls but did not significantly differ in thematic content, presence of tension, meaning-making, self-connections that reflect identity stability or identity change and affective responses to memory retrieval. Furthermore, among the patients' memories that refer to the TBI-related event itself, only one contained an explicit meaning. Despite the lack of specificity in memories, patients with severe TBI were able to extract meaning from personal memories although they could struggle with integrating the TBI-related event into their sense of identity. These characteristics of SDMs may contribute to disturbances in sense of self and continuity in patients with severe TBI, as well as difficulties in personal or social adjustment. These results also open up relevant prospects for psychological interventions in identity-related issues in patients with TBI.


Stimulation of group III and IV muscle afferents reflexly decreases total pulmonary resistance in dogs.
Although previous investigations have shown that stimulating group III and IV afferents reflexly decreased transverse tension from the trachealis muscle, these measurements provided no functional information about airway caliber. We therefore electrically stimulated gracilis muscle afferents in paralyzed, chloralose anesthetized dogs while recording total pulmonary resistance breath by breath. In addition, we recorded compound action potentials to determine which afferents were stimulated by current intensities of 3, 5, 20, 70 and 200 times motor threshold. We found that stimulating (20 Hz) the nerves at 3 times threshold, a current intensity which activated only group I and II afferents, had no effect on total pulmonary resistance, whereas stimulating the nerves at 5, 20 and 70 times threshold, current intensities which activated group I, II and III afferents, significantly decreased this variable. Stimulating the nerves at 200 times threshold, a current intensity which activated group IV as well as group I, II and III afferents, decreased total pulmonary resistance significantly more than did stimulating the nerves at 5, 20 or 70 times threshold. In addition stimulating the nerves at 200 times threshold but at frequencies of 2 and 5 Hz significantly decreased total pulmonary resistance. The decrease in total pulmonary resistance evoked by electrically stimulating the nerves at 200 times threshold was unaffected by propranolol but was abolished by atropine methylnitrate. We conclude that stimulating group III and IV gracilis muscle afferents in dogs reflexly decreases total pulmonary resistance, an effect due to the withdrawal of a tonic cholinergic input to the airways.


Spectrally reconfigurable integrated multi-spot particle trap.
Optical manipulation of small particles in the form of trapping, pushing, or sorting has developed into a vast field with applications in the life sciences, biophysics, and atomic physics. Recently, there has been increasing effort toward integration of particle manipulation techniques with integrated photonic structures on self-contained optofluidic chips. Here, we use the wavelength dependence of multi-spot pattern formation in multimode interference (MMI) waveguides to create a new type of reconfigurable, integrated optical particle trap. Interfering lateral MMI modes create multiple trapping spots in an intersecting fluidic channel. The number of trapping spots can be dynamically controlled by altering the trapping wavelength. This novel, spectral reconfigurability is utilized to deterministically move single and multiple particles between different trapping locations along the channel. This fully integrated multi-particle trap can form the basis of high throughput biophotonic assays on a chip.


Clinical yield of diagnostic endoscopic retrograde cholangiopancreatography in orthotopic liver transplant recipients with suspected biliary complications.
Diagnostic endoscopic retrograde cholangiopancreatography (D-ERCP) is commonly performed for the evaluation of biliary complications after orthotopic liver transplantation (OLT). This practice is contrary to the national trend of reserving endoscopic retrograde cholangiopancreatography (ERCP) for therapeutic purposes. Our aim was to evaluate the clinical yield and complications of D-ERCP in OLT recipients. In this retrospective study, 165 OLT recipients who underwent ERCP between January 2006 and December 2010 at the University of Michigan were divided into 2 groups: (1) a therapeutic endoscopic retrograde cholangiopancreatography (T-ERCP) group (if they met prespecified criteria that suggested a high likelihood of endoscopic intervention) and (2) a D-ERCP group (if there was clinical suspicion of biliary disease but they did not meet any criteria). The 2 groups were compared with respect to the proportion of subjects undergoing high-yield ERCP, which was defined as a procedure resulting in a clinically important intervention that modified the disease course. 66.3% of the D-ERCP procedures were classified as high-yield, whereas 90.1% of the T-ERCP procedures were (P < 0.001). Serious complications were infrequent in both groups. A survey of practitioners caring for OLT recipients suggested that the rate of high-yield D-ERCP seen in this study is congruent with what is considered acceptable in clinical practice. In conclusion, although T-ERCP is more likely to reveal a pathological process requiring an intervention, D-ERCP appears to be an acceptable clinical strategy for OLT recipients because of the high likelihood of a high-yield study and the low rate of serious complications.


Molecular subtyping scheme for serotypes HS1 and HS4 of Campylobacter jejuni.
We describe a molecular subtyping scheme for two principal O (heat-stable [HS]) serotypes of Campylobacter jejuni, HS1 and the HS4 complex. A 16S rRNA gene-specific probe confirmed that almost all the C. jejuni strains had three copies of this gene, and strains could be assigned with complete typeability to 1 of 16 combined (Pst1 and HaeIII) 16S ribotypes. Macrorestriction profiles (mrps) consisting of up to 10 SmaI fragments from approximately 40 to approximately 480 kbp were resolved by pulsed-field gel electrophoresis (PFGE). There were 11 mrps among the HS1 strains and 9 mrps among HS4 strains which corresponded to valid types--they occurred in multiple isolates, hosts, places, and times. There were 14 additional single-strain mrp fingerprints in HS1 and 20 in HS4. PFGE exhibited complete typeability when formaldehyde fixation of cells was employed, and PFGE was generally more differential than ribotyping. The data presented elucidate a high-resolution genotypic subtyping scheme for these common subspecific phenotypes of C. jejuni, which is both coherent and efficient for epidemiological purposes.


Imaging the assembly, structure and activity of type III secretion systems.
The type III secretion system (T3SS) is a sophisticated molecular machinery of Gram-negative bacteria used to 'inject' (translocate) bacterial proteins (effectors) into eukaryotic cells. For this, the T3SS has to assemble into a multiprotein complex, which is constituted of distinct parts; a basal body spanning the two bacterial membranes connected with a cytoplasmic bulb, an attached needle structure resembling a molecular syringe, and a distal needle tip structure that re-organizes into a 'translocon', which is a protein complex that inserts into the host cellular membrane. Upon engaging with eukaryotic cells, the T3SSs perform 'single-step' translocation of bacterial effector proteins across three membranes (two bacterial and one eukaryotic). Since the formulation of the major concepts of the T3SS about 15 years ago, imaging has been a major ingredient for elucidating the T3SS structure and function. Direct observation of molecular events in the context of cells will remain a key feature for better understanding of T3SS structure, regulation and function. In this review we describe how light and electron microscopy have been used to shed light on the processes of maturation and activity of the T3SS. Furthermore, we highlight recent imaging innovations with the potential to provide better insight into the T3SS structure and function.


Evaluation of a Novel Handoff Communication Strategy for Patients Admitted from the Emergency Department.
Miscommunication during inter-unit handoffs between emergency and internal medicine physicians may jeopardize patient safety. Our goal was to evaluate the impact of a structured communication strategy on the quality of admission handoffs. We conducted a mixed-methods, pre-test/post-test study at a 560-bed academic health center with 60,000 emergency department (ED) patient visits per year. Admission-handoff best practices were integrated into a modified SBAR format, resulting in the Situation, Background, Assessment, Responsibilities & Risk, Discussion & Disposition, Read-back & Record (SBAR-DR) model. Physician handoff conversations were recorded and transcribed for the 60 days before (n=110) and 60 days after (n=110) introduction of the SBAR-DR strategy. Transcriptions were scored by two blinded physicians using a 16-item scoring instrument. The primary outcome was the composite handoff quality score. We assessed physician perceptions via a post-intervention survey. The composite quality score improved in the post-intervention phase (7.57 + 2.42 vs. 8.45 + 2.51, p=.0085). Three of the 16 individual scoring elements also improved, including time for questions (70.6% vs. 82.7%, p=.0344) and confirmation of disposition plan (41.8% vs. 62.7%, p=.0019). The majority of emergency and internal medicine physicians felt that the SBAR-DR model had a positive impact on patient safety and handoff efficiency. Implementation of the SBAR-DR strategy resulted in improved verbal handoff quality. Agreement upon a clear disposition plan was the most improved element, which is of great importance in delineating responsibility of care and streamlining ED throughput. Future efforts should focus on nurturing broader physician buy-in to facilitate institution-wide implementation.


SADMFR guidelines for the use of Cone-Beam Computed Tomography/ Digital Volume Tomography.
Cone-Beam Computed Tomography (CBCT) has been introduced in 1998. This radiological imaging procedure has been provided for dentistry and is comparable to computed tomography (CT) in medicine. It is expected that CBCT will have the same success in dental diagnostic imaging as computed tomography had in medicine. Just as CT is responsible for a significant rise in radiation dose to the population from medical X-ray diagnostics, CBCT studies will be accompanied by a significant increase of the dose to our patients by dentistry. Because of the growing concern for an uncritical and consequently rapidly increasing use of CBCT the Swiss Society of Dentomaxillofacial Radiology convened a first consensus conference in 2011 to formulate indications for CBCT, which can be used as guidelines. In this meeting, oral and maxillofacial surgery, orthodontics and temporomandibular joint disorders and diseases were treated and the most important and most experienced users of DVT in these areas were asked to participate. In general, a highly restrictive use of CBCT is required. Justifying main criterion for CBCT application is that additional, therapy-relevant information is expected that should lead to a significant benefit in patient care. All users of CBCT should have completed a structured, high-level training, just like that offered by the Swiss Society of Dentomaxillofacial Radiology.


Reliability of a Standardized Mini-Mental State Examination compared with the traditional Mini-Mental State Examination.
The objective of this study was to compare the reliability of the Mini-Mental State Examination with that of a new Standardized Mini-Mental State Examination, which has expanded guidelines for administration and scoring. The subjects were 32 stable elderly residents of a nursing home and 16 elderly residents of a chronic care hospital unit. Six raters administered the Folstein Mini-Mental State to 22 of these stable elderly subjects, and five raters administered the standardized version to 26 of these subjects. Each subject was tested on three different occasions 1 week apart. Each rater tested 4-6 subjects at the first and third weeks and 4-6 different subjects at the second week. The analytic technique used was one-way analysis of variance to estimate the interrater variance and the intrarater variance. The intrarater variance on all occasions was reduced by 86% and the interrater variance was reduced by 76% when the Standardized Mini-Mental State was used; the reductions in variance were significant (p less than 0.003). The intraclass correlation for the Mini-Mental State was 0.69; for the standardized version it was 0.90. It took less time to administer the Standardized Mini-Mental State than the Mini-Mental State. The Standardized Mini-Mental State had better reliability than the Mini-Mental State in this study group. Although the improved reliability of the Standardized Mini-Mental State was achieved by reducing measurement noise, this advantage would likely occur in a broad spectrum of patients.


'I want to feel at home': establishing what aspects of environmental design are important to people with dementia nearing the end of life.
The design of environments in which people with dementia live should be understandable, reinforce personal identity and maintain their abilities. The focus on supporting people with dementia to live well has omitted considering the needs or wishes for a supportive physical environment of those who are nearing the end of their lives. Using a combination of focus groups and a Delphi survey, this study explored the views of people with dementia, family carers and professionals on what aspects of the physical environment would be important to support a good quality of life to the very end. Three focus groups were carried out in three cities along the East Coast of Australia using a discussion guide informed by a literature review. Focus groups comprised recently bereaved family carers of people with dementia (FG1), people with dementia and family carers of people with dementia (FG2) and practitioners caring for people with dementia nearing or at the end of their lives (FG3). Focus group conversations were audio-recorded with participants' consent. Audio files were transcribed verbatim and analysed thematically to identify environmental features that could contribute to achieving the goal of providing a comfortable life to the end. A list of design features derived from analysis of focus group transcripts was distributed to a range of experts in the dementia field and a consensus sought on their appropriateness. From this, a set of features to inform the design of environments for people with dementia nearing the end of life was defined. Eighteen people took part in three focus groups: two with dementia, eleven current or recently bereaved family carers and five practitioners. There were differences in opinion on what were important environmental considerations. People with dementia and family carers identified comfort through engagement, feeling at home, a calm environment, privacy and dignity and use of technology to remain connected as important. For practitioners, design to facilitate duty of care and institutional influences on their practice were salient themes. Twenty one experts in the dementia field took part in the survey to agree a consensus on the desirable features derived from analysis of focus group transcripts, with fifteen features agreed. The fifteen features are compatible with the design principles for people with dementia who are mobile, but include a stronger focus on sensory engagement. We suggest that considering these features as part of a continuum of care will support good practice and offer those with dementia the opportunity to live well until the end and give their families a more positive experience in the last days of their lives together.


Defence versus defence: Are crucian carp trading off immune function against predator-induced morphology?
Numerous species adopt inducible defence strategies; that is, they have phenotypically plastic traits that decrease the risk of capture and consumption by potential predators. The benefits of expressing alternative phenotypes in high- vs. low-risk environments are well documented. However, inducible anti-predator traits are also expected to incur costs, as they are not expressed when predators are absent, yet empirical evidence of such costs remains scarce. Virtually, all animals in nature are simultaneously under strong selection to evade both capture by predators and infection by parasites or pathogens and, hence, display a diverse arsenal of defences to combat these threats, raising the possibility of trade-offs between defences. A classic example of a predator-induced morphological defence is the deep-bodied shape of crucian carp that reduces risk of predation from gape-size-limited predators. The goal of this study was to examine whether predator exposure affects also immune function in crucian carp, and whether the degree of expressed morphological defence is traded off against immune function in individuals. Following exposure to manipulations of perceived risk (predator presence/absence) in a long-term experiment (8 months), key aspects of innate immune function and individual differences in the expression of inducible morphological defence were quantified. Predator-exposed individuals showed lower haptoglobin levels and complement activity, but higher natural antibody titres than fish from predator-free conditions. When experimentally challenged with a mimicked bacterial infection (LPS injection), fish reared in the presence of a natural predator showed a weaker immune response. Moreover, among predator-exposed individuals, the magnitude of morphological defence expression correlated with both baseline immune function and the ability to mount an immune response. However, these relationships were not consistently supportive of a general trade-off among defences. Our results suggest that fish exposed to predators on average reduce investment in immune function, and, further, the observed relationships among defences in predator-exposed individuals can best be explained from individual fitness and pace-of-life perspectives.


3-D-Printed Registration Phantom for Combined Ultrasound and Optical Imaging of Biological Tissues.
Efforts to develop quantitative ultrasound biomarkers would benefit from comparisons between ultrasound data and higher-resolution images of the tissue microstructure, such as from optical microscopy. However, only a few studies have used these methods for multiscale imaging because it is difficult to register low-resolution (>100 μm) ultrasound images to high-resolution microscopy images. To address this need, we have designed a 3-D-printed registration phantom that is made of a hard fluorescent resin, fits into a glass-bottom dish and can be used to calculate a coordinate system transform between ultrasound and optical microscopy. We report the phantom design, a registration protocol and an example registration using 18.5-MHz ultrasound and second harmonic generation microscopy. We evaluate the registration precision, achieving standard deviations smaller than the ultrasound resolution across all axes, and illustrate on a mouse mammary gland that this method yields results superior to those of manual landmark registration.


Daily co-trimoxazole prophylaxis in severely immunosuppressed HIV-infected adults in Africa started on combination antiretroviral therapy: an observational analysis of the DART cohort.
Co-trimoxazole prophylaxis can reduce mortality from untreated HIV infection in Africa; whether benefits occur alongside combination antiretroviral therapy (ART) is unclear. We estimated the effect of prophylaxis after ART initiation in adults. Participants in our observational analysis were from the DART randomised trial of management strategies in HIV-infected, symptomatic, previously untreated African adults starting triple-drug ART with CD4 counts lower than 200 cells per muL. Co-trimoxazole prophylaxis was not routinely used or randomly allocated, but was variably prescribed by clinicians. We estimated effects on clinical outcomes, CD4 cell count, and body-mass index (BMI) using marginal structural models to adjust for time-dependent confounding by indication. DART was registered, number ISRCTN13968779. 3179 participants contributed 14 214 years of follow-up (8128 [57%] person-years on co-trimoxazole). Time-dependent predictors of co-trimoxazole use were current CD4 cell count, haemoglobin concentration, BMI, and previous WHO stage 3 or 4 events on ART. Present prophylaxis significantly reduced mortality (odds ratio 0.65, 95% CI 0.50-0.85; p=0.001). Mortality risk reduction on ART was substantial to 12 weeks (0.41, 0.27-0.65), sustained from 12-72 weeks (0.56, 0.37-0.86), but not evident subsequently (0.96, 0.63-1.45; heterogeneity p=0.02). Variation in mortality reduction was not accounted for by time on co-trimoxazole or current CD4 cell count. Prophylaxis reduced frequency of malaria (0.74, 0.63-0.88; p=0.0005), an effect that was maintained with time, but we observed no effect on new WHO stage 4 events (0.86, 0.69-1.07; p=0.17), CD4 cell count (difference vs non-users, -3 cells per muL [-12 to 6]; p=0.50), or BMI (difference vs non-users, -0.04 kg/m(2) [-0.20 to 0.13); p=0.68]. Our results reinforce WHO guidelines and provide strong motivation for provision of co-trimoxazole prophylaxis for at least 72 weeks for all adults starting combination ART in Africa. UK Medical Research Council, the UK Department for International Development, the Rockefeller Foundation, GlaxoSmithKline, Gilead Sciences, Boehringer-Ingelheim, and Abbott Laboratories.


Scientific Rationale for the Inclusion and Exclusion Criteria for Intravenous Alteplase in Acute Ischemic Stroke: A Statement for Healthcare Professionals From the American Heart Association/American Stroke Association.
To critically review and evaluate the science behind individual eligibility criteria (indication/inclusion and contraindications/exclusion criteria) for intravenous recombinant tissue-type plasminogen activator (alteplase) treatment in acute ischemic stroke. This will allow us to better inform stroke providers of quantitative and qualitative risks associated with alteplase administration under selected commonly and uncommonly encountered clinical circumstances and to identify future research priorities concerning these eligibility criteria, which could potentially expand the safe and judicious use of alteplase and improve outcomes after stroke. Writing group members were nominated by the committee chair on the basis of their previous work in relevant topic areas and were approved by the American Heart Association Stroke Council's Scientific Statement Oversight Committee and the American Heart Association's Manuscript Oversight Committee. The writers used systematic literature reviews, references to published clinical and epidemiology studies, morbidity and mortality reports, clinical and public health guidelines, authoritative statements, personal files, and expert opinion to summarize existing evidence and to indicate gaps in current knowledge and, when appropriate, formulated recommendations using standard American Heart Association criteria. All members of the writing group had the opportunity to comment on and approved the final version of this document. The document underwent extensive American Heart Association internal peer review, Stroke Council Leadership review, and Scientific Statements Oversight Committee review before consideration and approval by the American Heart Association Science Advisory and Coordinating Committee. After a review of the current literature, it was clearly evident that the levels of evidence supporting individual exclusion criteria for intravenous alteplase vary widely. Several exclusionary criteria have already undergone extensive scientific study such as the clear benefit of alteplase treatment in elderly stroke patients, those with severe stroke, those with diabetes mellitus and hyperglycemia, and those with minor early ischemic changes evident on computed tomography. Some exclusions such as recent intracranial surgery are likely based on common sense and sound judgment and are unlikely to ever be subjected to a randomized, clinical trial to evaluate safety. Most other contraindications or warnings range somewhere in between. However, the differential impact of each exclusion criterion varies not only with the evidence base behind it but also with the frequency of the exclusion within the stroke population, the probability of coexistence of multiple exclusion factors in a single patient, and the variation in practice among treating clinicians.


The Intersections of Chronic Noncancer Pain: Culturally Diverse Perspectives on Disease Burden.
This study highlights the burden of chronic non-cancer-related pain from the perspectives of three culturally and linguistically diverse communities, using an intersectionality analysis. Specifically, we identify how multiple social identities intersect to account for the unequal distribution of the burden of chronic pain. Six focus groups of 41 culturally and linguistically diverse participants (Mandaean, Assyrian, and Vietnamese) living with chronic noncancer pain were conducted in South-West Sydney, Australia, between February and July 2015. Data were analyzed using inductive and intersectional methodology. The interaction between a patient with chronic pain from a culturally and linguistically diverse background and the health system is influenced by four identified social identities that interact to create relative positions of disadvantage for the patient within the health system and with health care providers. The social identities identified were ethnoculture, social class, migration status, and gender. Health care providers must consider how the intersectionality of social identities related to ethnoculture, social class, migration status, and gender can factor into the creation and maintenance of chronic pain disparities. A greater, more thoughtful incorporation of intersectionality in chronic pain research and clinical practice will ensure that pain management approaches are designed and applied in a way that reflects the social context of affected communities and individuals from those communities.


A genome-wide search for asthma susceptibility loci in ethnically diverse populations.
Asthma is an inflammatory airways disease associated with intermittent respiratory symptoms, bronchial hyper-responsiveness (BHR) and reversible airflow obstruction and is phenotypically heterogeneous. Patterns of clustering and segregation analyses in asthma families have suggested a genetic component to asthma. Previous studies reported linkage of BHR and atopy to chromosomes 5q (refs 7-9), 6p (refs 10-12), 11q (refs 13-15), 14q (ref. 16), and 12q (ref. 17) using candidate gene approaches. However, the relative roles of these genes in the pathogenesis of asthma or atopy are difficult to assess outside of the context of a genome-wide search. One genome-wide search in atopic sib pairs has been reported, however, only 12% of their subjects had asthma. We conducted a genome-wide search in 140 families with > or = 2 asthmatic sibs, from three racial groups and report evidence for linkage to six novel regions: 5p15 (P = 0.0008) and 17p11.1-q11.2 (P = 0.0015) in African Americans; 11p15 (P = 0.0089) and 19q13 (P = 0.0013) in Caucasians; 2q33 (P = 0.0005) and 21q21 (P = 0.0040) in Hispanics. Evidence for linkage was also detected in five regions previously reported to be linked to asthma-associated phenotypes: 5q23-31 (P = 0.0187), 6p21.3-23 (P = 0.0129), 12q14-24.2 (P = 0.0042), 13q21.3-qter (P = 0.0014), and 14q11.2-13 (P = 0.0062) in Caucasians and 12q14-24.2 (P = 0.0260) in Hispanics.


Process evaluation in a multisite, primary obesity-prevention trial in American Indian schoolchildren.
We describe the development, implementation, and use of the process evaluation component of a multisite, primary obesity prevention trial for American Indian schoolchildren. We describe the development and pilot testing of the instruments, provide some examples of the criteria for instrument selection, and provide examples of how process evaluation results were used to document and refine intervention components. The theoretical and applied framework of the process evaluation was based on diffusion theory, social learning theory, and the desire for triangulation of multiple modes of data collection. The primary objectives of the process evaluation were to systematically document the training process, content, and implementation of 4 components of the intervention. The process evaluation was developed and implemented collaboratively so that it met the needs of both the evaluators and those who would be implementing the intervention components. Process evaluation results revealed that observation and structured interviews provided the most informative data; however, these methods were the most expensive and time consuming and required the highest level of skill to undertake. Although the literature is full of idealism regarding the uses of process evaluation for formative and summative purposes, in reality, many persons are sensitive to having their work evaluated in such an in-depth, context-based manner as is described. For this reason, use of structured, quantitative, highly objective tools may be more effective than qualitative methods, which appear to be more dependent on the skills and biases of the researcher and the context in which they are used.


The effects of placental extract on fibroblast proliferation.
Human placental extract is used in the treatment of skin wrinkles and wounds. To date, no studies have evaluated the effects of placental extract on dermal fibroblast proliferation. To investigate the effects of placental extract versus ascorbic acid on fibroblast proliferation and transforming growth factor (TGF)-beta1 expression, cultured human fibroblasts were treated with placental extract (0, 0.08, 0.16, 0.32, and 0.64%) or L-ascorbic acid-2-phosphate magnesium (0, 0.01, 0.1, 1.0, and 10 mM). Fibroblast proliferation was determined by MTT assay, and TGF-beta1 protein expression was analyzed by ELISA. The proliferation of fibroblasts increased significantly after treatment with placental extract at concentrations of 0.32 and 0.64% and with L-ascorbic acid-2-phosphate magnesium at concentrations of 1.0 and 10 mM. Placental extract demonstrated no significant effects on TGF-beta1 expression; however, TGF-beta1 expression significantly increased after treatment with ascorbic acid at concentrations of 1.0 and 10 mM. Placental extract and ascorbic acid had similar effects on fibroblast proliferation; however, placental extract did not significantly increase TGF-beta1 protein expression.


4Pipe4--A 454 data analysis pipeline for SNP detection in datasets with no reference sequence or strain information.
Next-generation sequencing datasets are becoming more frequent, and their use in population studies is becoming widespread. For non-model species, without a reference genome, it is possible from a panel of individuals to identify a set of SNPs that can be used for further population genotyping. However the lack of a reference genome to which the sequenced data could be compared makes the finding of SNPs more troublesome. Additionally when the data sources (strains) are not identified (e.g. in datasets of pooled individuals), the problem of finding reliable variation in these datasets can become much more difficult due to the lack of specialized software for this specific task. Here we describe 4Pipe4, a 454 data analysis pipeline particularly focused on SNP detection when no reference or strain information is available. It uses a command line interface to automatically call other programs, parse their outputs and summarize the results. The variation detection routine is built-in in the program itself. Despite being optimized for SNP mining in 454 EST data, it is flexible enough to automate the analysis of genomic data or even data from other NGS technologies. 4Pipe4 will output several HTML formatted reports with metrics on many of the most common assembly values, as well as on all the variation found. There is also a module available for finding putative SSRs in the analysed datasets. This program can be especially useful for researchers that have 454 datasets of a panel of pooled individuals and want to discover and characterize SNPs for subsequent individual genotyping with customized genotyping arrays. In comparison with other SNP detection approaches, 4Pipe4 showed the best validation ratio, retrieving a smaller number of SNPs but with a considerably lower false positive rate than other methods. 4Pipe4's source code is available at https://github.com/StuntsPT/4Pipe4.


'Equivalence' and the translation and adaptation of health-related quality of life questionnaires.
The increasing use of health-related quality of life (HRQOL) questionnaires in multinational studies has resulted in the translation of many existing measures. Guidelines for translation have been published, and there has been some discussion of how to achieve and assess equivalence between source and target questionnaires. Our reading in this area had led us, however, to the conclusion that different types of equivalence were not clearly defined, and that a theoretical framework for equivalence was lacking. To confirm this we reviewed definitions of equivalence in the HRQOL literature on the use of generic questionnaires in multicultural settings. The literature review revealed: definitions of 19 different types of equivalence; vague or conflicting definitions, particularly in the case of conceptual equivalence; and the use of many redundant terms. We discuss these findings in the light of a framework adapted from cross-cultural psychology for describing three different orientations to cross-cultural research: absolutism, universalism and relativism. We suggest that the HRQOL field has generally adopted an absolutist approach and that this may account for some of the confusion in this area. We conclude by suggesting that there is an urgent need for a standardized terminology within the HRQOL field, by offering a standard definition of conceptual equivalence, and by suggesting that the adoption of a universalist orientation would require substantial changes to guidelines and more empirical work on the conceptualization of HRQOL in different cultures.


Structural changes within trophic levels are constrained by within-family assembly rules at lower trophic levels.
Historical contingency broadly refers to the proposition that even random historical events can constrain the ecological and evolutionary pathways of organisms and that of entire communities. Focusing on communities, these pathways can be reflected into specific structural changes within and across trophic levels - how species interact with and affect each other - which has important consequences for species coexistence. Using the registry of the last 2000 years of plant introductions and their novel herbivores encountered in Central Europe, we find that the order of arrival of closely related (but not of distantly related) plant species constrained the structural changes within the trophic level formed by herbivore species across the observation period. Because it is difficult for field and lab experiments to be conducted over hundreds of years to record and replay the assembly history of a community, our study provides an alternative to understand how structural changes have occurred across extensive periods of time.


Opioid-induced androgen deficiency (OPIAD).
Opioid therapy is one of the most effective forms of analgesia currently in use. In the past few decades, the use of opioids as a long-term treatment for chronic pain has increased dramatically. Accompanying this upsurge in the use of long-term opioid therapy has been an increase in the occurrence of opioid associated endocrinopathy, most commonly manifested as an androgen deficiency and therefore referred to as opioid associated androgen deficiency (OPIAD). This syndrome is characterized by the presence of inappropriately low levels of gonadotropins (follicle stimulating hormone and luteinizing hormone) leading to inadequate production of sex hormones, particularly testosterone. Symptoms that may manifest in patients with OPIAD include reduced libido, erectile dysfunction, fatigue, hot flashes, and depression. Physical findings may include reduced facial and body hair, anemia, decreased muscle mass, weight gain, and osteopenia or osteoporosis. Additionally, both men and women with OPIAD may suffer from infertility. While the literature regarding OPIAD remains limited, it is apparent that OPIAD is becoming increasingly prevalent among chronic opioid consumers but often goes unrecognized. OPIAD can have a significant negative impact on the the quality of life of opioid users, and clinicians should anticipate the potential for its occurrence whenever long-term opioid prescribing is undertaken. Once diagnosed, treatment for OPIAD may be offered utilizing a number of androgen replacement therapy options including a variety of testosterone preparations and, for female patients with OPIAD, dehydroepiandrosterone (DHEA) supplementation. Follow-up evaluation of patients receiving androgen replacement therapy should include a review of any unresolved symptoms of hypogonadism, laboratory evaluation, and surveillance for potential adverse effects of androgen replacement therapy including prostate disease in males.:


[Neuromuscular blockers in Brazil.].
There are no statistical data on the use of neuromuscular blockers in Brazil. This study aimed at statistically analyzing this topic. Our study has compiled 831 answers to a questionnaire filled by anesthesiologists attending the 48th Brazilian Congress of Anesthesiology in Recife, 2001, and via Internet by anesthesiologists whose e-mail addresses are in the Brazilian Society of Anesthesiology web page (www.sba.com.br). The following data were evaluated: years of experience with the specialty, region where anesthesiologists practice, neuromuscular blockers (NMB) usage in order of preference, indications for succinylcholine, neuromuscular transmission monitor usage, blockade recovery criteria, neostigmine usage, NMB administration routes and description of observed complications. Most anesthesiologists practice for more than 11 years and the highest number of answers have come from the Southeastern region of Brazil. Most common NMB is atracurium, followed by pancuronium and succinylcholine. Succinylcholine is more frequently used for rapid sequence induction and in children (80% and 25%, respectively). Neuromuscular transmission monitors are never used by 53% of anesthesiologists, and 92% of them use clinical signs as blockade recovery criteria. Neostigmine is routinely used by 45% of professionals and 94% of them administer NMB in bolus. Approximately 30% have referred NMB-related complications. Most frequent complications were prolonged blockade, severe bronchospasm and residual curarization. Atracurium is the most popular neuromuscular blocker in Brazil; there is a high percentage of succinylcholine usage in non-emergency situations; neuromuscular transmission monitors are seldom used and, as a corollary, there is a significant percentage of clinical criteria to consider patients recovered. We have observed that approximately 30% of anesthesiologists had some type of NMB-related complication.


High-Q, low index-contrast polymeric photonic crystal nanobeam cavities.
We present the design, fabrication and characterization of high-Q (Q=36,000) polymeric photonic crystal nanobeam cavities made of two polymers that have an ultra-low index contrast (ratio=1.15) and observed thermo-optical bistability at hundred microwatt power level. Due to the extended evanescent field and small mode volumes, polymeric nanobeam cavities are ideal platform for ultra-sensitive biochemical sensing. We demonstrate that these sensors have figures of merit (FOM=9190) two orders of magnitude greater than surface plasmon resonance based sensors, and outperform the commercial Biacore(TM) sensors. The demonstration of high-Q cavity in low-index-contrast polymers can open up versatile applications using a broad range of functional and flexible polymeric materials.


Cerebral vascular structure in the motor cortex of adult mice is stable and is not altered by voluntary exercise.
The cerebral vasculature provides blood flow throughout the brain, and local changes in blood flow are regulated to match the metabolic demands of the active brain regions. This neurovascular coupling is mediated by real-time changes in vessel diameter and depends on the underlying vascular network structure. Neurovascular structure is configured during development by genetic and activity-dependent factors. In adulthood, it can be altered by experiences such as prolonged hypoxia, sensory deprivation and seizure. Here, we have sought to determine whether exercise could alter cerebral vascular structure in the adult mouse. We performed repeated in vivo two-photon imaging in the motor cortex of adult transgenic mice expressing membrane-anchored green fluorescent protein in endothelial cells (tyrosine endothelial kinase 2 receptor (Tie2)-Cre:mTmG). This strategy allows for high-resolution imaging of the vessel walls throughout the lifespan. Vascular structure, as measured by capillary branch point number and position, segment diameter and length remained stable over a time scale of months as did pericyte number and position. Furthermore, we compared the vascular structure before, during, and after periods of voluntary wheel running and found no alterations in these same parameters. In both running and control mice, we observed a low rate of capillary segment subtraction. Interestingly, these rare subtraction events preferentially remove short vascular loops.


VEGF internalization is not required for VEGFR-2 phosphorylation in bioengineered surfaces with covalently linked VEGF.
Vascular endothelial growth factor (VEGF) is known to activate proliferation, migration, and survival pathways in endothelial cells through phosphorylation of VEGF receptor-2 (VEGFR-2). VEGF has been incorporated into biomaterials through encapsulation, electrostatic sequestration, and covalent attachment, but the effect of these immobilization strategies on VEGF signaling has not been thoroughly investigated. Further, although growth factor internalization along with the receptor generally occurs in a physiological setting, whether this internalization is needed for receptor phosphorylation is not entirely clear. Here we show that VEGF covalently bound through a modified heparin molecule elicits an extended response of pVEGFR-2 in human umbilical vein endothelial cells (HUVECs) and that the covalent linkage reduces internalization of the growth factor during receptor endocytosis. Optical tweezer measurements show that the rupture force required to disrupt the heparin-VEGF-VEGFR-2 interaction increases from 3-8 pN to 6-12 pN when a covalent bond is introduced between VEGF and heparin. Importantly, by covalently binding VEGF to a heparin substrate, the stability (half-life) of VEGF is extended over three-fold. Here, mathematical models support the biological conclusions, further suggesting that VEGF internalization is significantly reduced when covalently bound, and indicating that VEGF is available for repeated phosphorylation events.


Effects of Caffeine on Tennis Serve Accuracy.
This study examined the effects of caffeine on tennis serve accuracy. Division II tennis athletes (n = 10) completed two serve trials (double-blind, counterbalanced) following ingestion of 6 mg/kg of caffeine or matched placebo an hour prior to data collection. During each tennis serve trial, participants completed 48 non-fault serves divided into 3 sets with 2 serves per 8 different targets. Following each 2 serves per target format, participants completed a shuttle run sprint. Separate 2 (trial) × 8 (targets) repeated measures analysis of variances (ANOVAs) were used for distances from the target center "delta", and necessary tries for each of the 3 sets. A 2 (treatment) × 3 (set) repeated measures ANOVA was used for shuttle run times. While results were not significant, the treatment main effect approached significance (p = 0.07) in set 2 for the delta of distances when comparing caffeine (96.2 cm ± 19.8) versus placebo (107.1 cm ± 16.3). While there was no significance in sprint times, each sprint was consistently faster following caffeine consumption. Post-trial surveys revealed subjective responses approached significance with greater feelings of stomach distress (p = 0.08) and nervousness (p = 0.13) following caffeine and elevated feeling of fatigue (p = 0.19) following placebo. Therefore, with no impairment in serve accuracy coupled with some evidence of reduced fatigue, results suggest caffeine may benefit tennis athletes. Extending the understanding of the effects of caffeine on tennis serve accuracy and performance could benefit overall match performance, with the potential of improving the match outcome in extended playtime.


Relative roles of cardiac and arterial baroreceptors in vasopressin regulation during hemorrhage in conscious dogs.
To determine the relative roles of cardiac and sinoaortic baroreceptors in mediation of arginine vasopressin (AVP) release, hemorrhage was performed in five groups of conscious splenectomized dogs: 1) all nerves intact; 2) either chronic surgical or acute pharmacological (intrapericardial lidocaine) cardiac denervation (CD); 3) chronic sinoarotic denervation (SAD); 4) combined chronic sinoartic denervation plus either chronic or acute cardiac denervation (SAD + CD); and 5) all nerves intact, but with ganglionic blockade. Hemorrhage (0.5 ml/min/kg) reduced mean arterial pressure similarly in the intact and CD groups. Decreases in mean arterial pressure were augmented in SAD, SAD + CD, and ganglion-blocked groups compared with responses in intact and CD groups. There were no differences in responses of plasma AVP to hemorrhage in the intact and CD groups, but the AVP response was significantly blunted in the SAD + CD group as compared with SAD alone. When compared during the early stage of hemorrhage, at the same reduction in mean arterial pressure, the rise in AVP was greater in the ganglion-blocked group than in the SAD + CD group, but was less than in the intact group. With a protocol to reduce mean arterial pressure by 20 mm Hg over the same period (42 +/- 0.6 minutes) in four of the groups, the blood volume required to reduce mean arterial pressure by 20 mm Hg was similar in the intact (20 +/- 1 ml/kg) and CD (21 +/- 1 ml/kg) groups, but was less in the SAD (12 +/- 1 ml/kg) and SAD + CD (12 +/- 1 ml/kg) groups. Again, similar increases were observed in AVP between the intact (50 +/- 9 pg/ml) and CD (51 +/- 9 pg/ml) groups, whereas increases in AVP were diminished in the SAD (11 +/- 3 pg/ml) and SAD + CD (7 +/- 2 pg/ml) groups. In the presence of an AVP antagonist, decreases in mean arterial pressure and increases in total peripheral resistance with hemorrhage were affected similarly in both the intact and CD groups, whereas hemodynamic impairment by AVP blockade was less marked in the SAD and SAD + CD groups. These results indicate that cardiac receptors are not the major regulators of AVP release during progressive hemorrhage in conscious dogs. However, when the complicating influences of sinoaortic reflexes were eliminated, a modest role for cardiac receptors was uncovered.


Broadly neutralizing immune responses against hepatitis C virus induced by vectored measles viruses and a recombinant envelope protein booster.
Hepatitis C virus (HCV) infection remains a serious public health problem worldwide. Treatments are limited, and no preventive vaccine is available. Toward developing an HCV vaccine, we engineered two recombinant measles viruses (MVs) expressing structural proteins from the prototypic HCV subtype 1a strain H77. One virus directs the synthesis of the HCV capsid (C) protein and envelope glycoproteins (E1 and E2), which fold properly and form a heterodimer. The other virus expresses the E1 and E2 glycoproteins separately, with each one fused to the cytoplasmic tail of the MV fusion protein. Although these hybrid glycoproteins were transported to the plasma membrane, they were not incorporated into MV particles. Immunization of MV-susceptible, genetically modified mice with either vector induced neutralizing antibodies to MV and HCV. A boost with soluble E2 protein enhanced titers of neutralizing antibody against the homologous HCV envelope. In animals primed with MV expressing properly folded HCV C-E1-E2, boosting also induced cross-neutralizating antibodies against two heterologous HCV strains. These results show that recombinant MVs retain the ability to induce MV-specific humoral immunity while also eliciting HCV neutralizing antibodies, and that anti-HCV immunity can be boosted with a single dose of purified E2 protein. The use of MV vectors could have advantages for pediatric HCV vaccination.


Embryonic precursor cells that express Trk receptors: induction of different cell fates by NGF, BDNF, NT-3, and CNTF.
Epidermal growth factor (EGF)-treated neurosphere cultures from embryonal striatum contain multipotential cells capable of neuronal, astrocytic, and oligodendroglial differentiation. In this study, we tested whether these neural precursor cells differentiate in the presence of neurotrophic factors. We first assayed neurosphere cells for expression of neurotrophin receptors. TrkA, TrkB, TrkC, and gp75 were detected by immunofluorescence microscopy in 60-80% of cells. In addition, the ciliary neurotrophic factor receptor alpha was expressed in 50-60% of cells. In the presence of the mitogen, EGF, treatment of stem cells with neurotrophic factors had no apparent effect. Removal of EGF from cells resulted in cessation of cell proliferation and pronounced astrocytic (glial fibrillary acidic protein+) differentiation. Neuronal (neurofilament+) and oligodendroglial (galactocerebroside+) cells appeared in cultures treated with neurotrophic factors. Nerve growth factor (NGF) resulted in bipolar neuronal cells, and brain-derived neurotrophic factor led to multipolar neuronal cells. Treatment with neurotrophin-3 or ciliary neurotrophic factor resulted in bipolar neuronal cells and oligodendrocytes. Neuronal differentiation in the presence of NGF was enhanced by extracellular matrix, and the resulting neuronal cells expressed choline acetyltransferase and, to a lesser degree, tyrosine hydroxylase. These studies demonstrate that neurotrophic factors influence the fates of these multipotential precursor cells. Indeed, the true utility of multipotential precursor cells is the production of different types of cells in different situations. Local cues, such as neurotrophic factors and extracellular matrix, may regulate production of different types of neural cells during development or in response to other stimuli, such as injury.


Surface binding of alamethicin stabilizes its helical structure: molecular dynamics simulations.
Alamethicin is an amphipathic alpha-helical peptide that forms ion channels. An early event in channel formation is believed to be the binding of alamethicin to the surface of a lipid bilayer. Molecular dynamics simulations are used to compare the structural and dynamic properties of alamethicin in water and alamethicin bound to the surface of a phosphatidylcholine bilayer. The bilayer surface simulation corresponded to a loosely bound alamethicin molecule that interacted with lipid headgroups but did not penetrate the hydrophobic core of the bilayer. Both simulations started with the peptide molecule in an alpha-helical conformation and lasted 2 ns. In water, the helix started to unfold after approximately 300 ps and by the end of the simulation only the N-terminal region of the peptide remained alpha-helical and the molecule had collapsed into a more compact form. At the surface of the bilayer, loss of helicity was restricted to the C-terminal third of the molecule and the rod-shaped structure of the peptide was retained. In the surface simulation about 10% of the peptide/water H-bonds were replaced by peptide/lipid H-bonds. These simulations suggest that some degree of stabilization of an amphipathic alpha-helix occurs at a bilayer surface even without interactions between hydrophobic side chains and the acyl chain core of the bilayer.


Influence of pediatric vaccines on amygdala growth and opioid ligand binding in rhesus macaque infants: A pilot study.
This longitudinal, case-control pilot study examined amygdala growth in rhesus macaque infants receiving the complete US childhood vaccine schedule (1994-1999). Longitudinal structural and functional neuroimaging was undertaken to examine central effects of the vaccine regimen on the developing brain. Vaccine-exposed and saline-injected control infants underwent MRI and PET imaging at approximately 4 and 6 months of age, representing two specific timeframes within the vaccination schedule. Volumetric analyses showed that exposed animals did not undergo the maturational changes over time in amygdala volume that was observed in unexposed animals. After controlling for left amygdala volume, the binding of the opioid antagonist [(11)C]diprenorphine (DPN) in exposed animals remained relatively constant over time, compared with unexposed animals, in which a significant decrease in [(11)C]DPN binding occurred. These results suggest that maturational changes in amygdala volume and the binding capacity of [(11)C]DPN in the amygdala was significantly altered in infant macaques receiving the vaccine schedule. The macaque infant is a relevant animal model in which to investigate specific environmental exposures and structural/functional neuroimaging during neurodevelopment.


Single Versus Multiple Solid Organ Injuries Following Blunt Abdominal Trauma.
We aimed to describe the pattern of solid organ injuries (SOIs) and analyze the characteristics, management and outcomes based on the multiplicity of SOIs. A retrospective study in a Level 1 trauma center was conducted and included patients admitted with blunt abdominal trauma between 2011 and 2014. Data were analyzed and compared for patients with single versus multiple SOIs. A total of 504 patients with SOIs were identified with a mean age of 28 ± 13 years. The most frequently injured organ was liver (45%) followed by spleen (30%) and kidney (18%). One-fifth of patients had multiple SOIs, of that 87% had two injured organs. Patients with multiple SOIs had higher frequency of head injury and injury severity scores (p < 0.05). The majority of SOIs were treated nonoperatively, whereas operative management was required in a quarter of patients, mostly in patients with multiple SOIs (p = 0.01). Blood transfusion, sepsis and hospital stay were greater in multiple than single SOIs (p < 0.05). The overall mortality was 11% which was comparable between the two groups. In patients with single SOIs, the mortality was significantly higher in those who had pancreatic (28.6%) or hepatic injuries (13%) than the other SOIs. SOIs represent one-tenth of trauma admissions in Qatar. Although liver was the most frequently injured organ, the rate of mortality was higher in pancreatic injury. Patients with multiple SOIs had higher morbidity which required frequent operative management. Further prospective studies are needed to develop management algorithm based on the multiplicity of SOIs.


Infanticide and cannibalism by male chimpanzees at Ngogo, Kibale National Park, Uganda.
Researchers have documented infanticide by adult males in four wild chimpanzee populations. Males in three of these have killed infants from outside of their own communities, but most infanticides, including one from Kanyawara, in Kibale National Park, Uganda, took place within communities. Here we report two new cases of infanticide by male chimpanzees at a second Kibale site, Ngogo, where the recently habituated chimpanzee community is the largest yet known. Both infanticides happended during boundary patrols, which occur at a high frequency there. Patrolling males attacked solitary females who were unable to defend their infants successfully. The victims were almost certainly not members of the Ngogo community. Males cannibalized both infants and completely consumed their carcasses. These observations show that infanticide by males is widespread in the Kibale population and that between-community infanticide also happens there. We discuss our observations in the context of the sexual selection hypothesis and other proposed explanations for infanticide by male chimpanzees. The observations support the arguments that infanticide has been an important selective force in chimpanzee social evolution and that females with dependent infants can be at great risk near range boundaries, but why male chimpanzees kill infants is still uncertain.


SacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformation.
Agrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transformation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce plant transformation efficiency. The SacB-SacR proteins are toxic to most Agrobacterium tumefaciens strains when they are grown on culture medium supplemented with sucrose. Therefore, SacB-SacR genes can be used as negative selection markers to suppress the overgrowth of A. tumefaciens in the plant tissue culture process. We generated a mutant A. tumefaciens strain GV2260 (recA-SacB/R) that has the SacB-SacR cassette inserted into the bacterial genome at the recA gene locus. The mutant Agrobacterium strain is sensitive to sucrose but maintains its ability to transform plant cells in both transient and stable transformation assays. We demonstrated that the mutant strain GV2260 (recA-SacB/R) can be inhibited by sucrose that reduces the overgrowth of Agrobacterium and therefore improves the plant transformation efficiency. We employed GV2260 (recA-SacB/R) to generate stable transgenic N. benthamiana plants expressing a CRISPR-Cas9 for knocking out a WRKY transcription factor.


Reforming the Swedish pharmaceuticals market: consequences for costs per defined daily dose.
In 2009 and 2010, the Swedish pharmaceuticals market was reformed. One of the stated policy goals was to achieve low costs for pharmaceutical products dispensed in Sweden. We use price and sales data for off-patent brand-name and generic pharmaceuticals to estimate a log-linear regression model, allowing us to assess how the policy changes affected the cost per defined daily dose. The estimated effect is an 18 % cost reduction per defined daily dose at the retail level and a 34 % reduction in the prices at the wholesale level (pharmacies' purchase prices). The empirical results suggest that the cost reductions were caused by the introduction of a price cap, an obligation to dispense the lowest-cost generic substitute available in the whole Swedish market, and the introduction of well-defined exchange groups. The reforms thus reduced the cost per defined daily dose for consumers while being advantageous also for the pharmacies, who saw their retail margins increase. However, pharmaceutical firms supplying off-patent pharmaceuticals experienced a clear reduction in the price received for their products.


Oxidative stress pathways in noncancerous human liver tissue to predict hepatocellular carcinoma recurrence: a prospective, multicenter study.
The prediction of cancer recurrence holds the key to improvement of the postoperative prognosis of patients. In this study, the recurrence of early-stage hepatocellular carcinoma (HCC) after curative hepatectomy was analyzed by the genome-wide gene-expression profiling on cancer tissue and the noncancerous liver tissue. Using the training set of 78 cases, the cytochrome P450 1A2 (CYP1A2) gene in noncancerous liver tissue was identified as the predictive candidate for postoperative recurrence (hazard ratio [HR], 0.447; 95% confidence interval [CI], 0.249-0.808; P = 0.010). Multivariate analysis revealed the statistically significant advantage of CYP1A2 down-regulation to predict recurrence (odds ratio, 0.534; 95% CI, 0.276-0.916; P = 0.036), and the expression of CYP1A2 protein was confirmed immunohistochemically. An independently multi-institutional cohort of 211 patients, using tissue microarrays, validated that loss of expression of CYP1A2 in noncancerous liver tissue as the only predictive factor of recurrence after curative hepatectomy for early-stage HCC (HR, 0.480; 95% CI, 0.256-0.902; P = 0.038). Gene set-enrichment analysis revealed close association of CYP1A2 down-regulation with oxidative stress pathways in liver tissue (P < 0.001, false discovery rate [FDR] = 0.042; P = 0.006, FDR = 0.035). Our results indicate these pathways as the molecular targets to prevent recurrence, as well as the potential prediction of the super high-risk population of HCC using liver tissue.


The global distribution and spread of the mobilized colistin resistance gene mcr-1.
Colistin represents one of the few available drugs for treating infections caused by carbapenem-resistant Enterobacteriaceae. As such, the recent plasmid-mediated spread of the colistin resistance gene mcr-1 poses a significant public health threat, requiring global monitoring and surveillance. Here, we characterize the global distribution of mcr-1 using a data set of 457 mcr-1-positive sequenced isolates. We find mcr-1 in various plasmid types but identify an immediate background common to all mcr-1 sequences. Our analyses establish that all mcr-1 elements in circulation descend from the same initial mobilization of mcr-1 by an ISApl1 transposon in the mid 2000s (2002-2008; 95% highest posterior density), followed by a marked demographic expansion, which led to its current global distribution. Our results provide the first systematic phylogenetic analysis of the origin and spread of mcr-1, and emphasize the importance of understanding the movement of antibiotic resistance genes across multiple levels of genomic organization.


The Importance of Gender-Related Anticancer Research on Mitochondrial Regulator Sodium Dichloroacetate in Preclinical Studies In Vivo.
Sodium dichloroacetate (DCA) is an investigational medicinal product which has a potential anticancer preparation as a metabolic regulator in cancer cells' mitochondria. Inhibition of pyruvate dehydrogenase kinases by DCA keeps the pyruvate dehydrogenase complex in the active form, resulting in decreased lactic acid in the tumor microenvironment. This literature review displays the preclinical research data on DCA's effects on the cell pyruvate dehydrogenase deficiency, pyruvate mitochondrial oxidative phosphorylation, reactive oxygen species generation, and the Na+-K+-2Cl- cotransporter expression regulation in relation to gender. It presents DCA pharmacokinetics and the hepatocarcinogenic effect, and the safety data covers the DCA monotherapy efficacy for various human cancer xenografts in vivo in male and female animals. Preclinical cancer researchers report the synergistic effects of DCA combined with different drugs on cancer by reversing resistance to chemotherapy and promoting cell apoptosis. Researchers note that female and male animals differ in the mechanisms of cancerogenesis but often ignore studying DCA's effects in relation to gender. Preclinical gender-related differences in DCA pharmacology, pharmacological mechanisms, and the elucidation of treatment efficacy in gonad hormone dependency could be relevant for individualized therapy approaches so that gender-related differences in treatment response and safety can be proposed.


Competing quantum effects in the free energy profiles and diffusion rates of hydrogen and deuterium molecules through clathrate hydrates.
Clathrate hydrates hold considerable promise as safe and economical materials for hydrogen storage. Here we present a quantum mechanical study of H2 and D2 diffusion through a hexagonal face shared by two large cages of clathrate hydrates over a wide range of temperatures. Path integral molecular dynamics simulations are used to compute the free-energy profiles for the diffusion of H2 and D2 as a function of temperature. Ring polymer molecular dynamics rate theory, incorporating both exact quantum statistics and approximate quantum dynamical effects, is utilized in the calculations of the H2 and D2 diffusion rates in a broad temperature interval. We find that the shape of the quantum free-energy profiles and their height relative to the classical free energy barriers at a given temperature, as well as the rate of diffusion, are strongly affected by competing quantum effects: above 25 K, zero-point energy (ZPE) perpendicular to the reaction path for diffusion between cavities decreases the quantum rate compared to the classical rate, whereas at lower temperatures tunneling outcompetes the ZPE and as a result the quantum rate is greater than the classical rate.


The Korean Cough Guideline: Recommendation and Summary Statement.
Cough is one of the most common symptom of many respiratory diseases. The Korean Academy of Tuberculosis and Respiratory Diseases organized cough guideline committee and cough guideline was developed by this committee. The purpose of this guideline is to help clinicians to diagnose correctly and treat efficiently patients with cough. In this article, we have stated recommendation and summary of Korean cough guideline. We also provided algorithm for acute, subacute, and chronic cough. For chronic cough, upper airway cough syndrome (UACS), cough variant asthma (CVA), and gastroesophageal reflux disease (GERD) should be considered. If UACS is suspicious, first generation anti-histamine and nasal decongestant can be used empirically. In CVA, inhaled corticosteroid is recommended in order to improve cough. In GERD, proton pump inhibitor is recommended in order to improve cough. Chronic bronchitis, bronchiectasis, bronchiolitis, lung cancer, aspiration, angiotensin converting enzyme inhibitor, habit, psychogenic cough, interstitial lung disease, environmental and occupational factor, tuberculosis, obstructive sleep apnea, peritoneal dialysis, and idiopathic cough can be also considered as cause of chronic cough. Level of evidence for treatment is mostly low. Thus, in this guideline, many recommendations are based on expert opinion. Further study regarding treatment for cough is mandatory.


Seroprevalence and risk factors associated with Helicobacter pylori infection in blood donors in Salvador, Northeast-Brazil.
Helicobacter pylori plays an important role in the etiology of peptic ulcer disease. Its prevalence appears to be higher in developing countries. We evaluated the seroprevalence of H. pylori and risk factors associated with infection in voluntary blood donors who attended the main blood center of the city of Salvador, Brazil. The subjects responded to an epidemiological questionnaire, with information about sex, age, race, lifestyle, social-economic level indicators, and residence and hygiene conditions. Anti-H. pylori antibody was determined by ELISA (Cobas Core, Roche). Three hundred and seven subjects were included in the study. Anti-H. pylori antibody results were indeterminate in 33 individuals (10.8%), who were excluded from analysis. Among the remaining 274 subjects, 187 (68.2%) were anti-H. pylori positive. Based on multivariate logistic regression analysis three variables were found to be significantly associated with a higher prevalence of H. pylori infection: absence of plumbing in the residence during childhood, a history of rainwater invading the dwelling during childhood, and low ingestion of milk.


Reverse vaccinology and subtractive genomics approaches for identifying common therapeutics against Mycobacterium leprae and Mycobacterium lepromatosis.
Mycobacterium leprae and Mycobacterium lepromatosis are gram-positive bacterial pathogens and the causative agents of leprosy in humans across the world. The elimination of leprosy cannot be achieved by multidrug therapy alone, and highlights the need for new tools and drugs to prevent the emergence of new resistant strains. In this study, our contribution includes the prediction of vaccine targets and new putative drugs against leprosy, using reverse vaccinology and subtractive genomics. Six strains of Mycobacterium leprae and Mycobacterium lepromatosis (4 and 2 strains, respectively) were used for comparison taking Mycobacterium leprae strain TN as the reference genome. Briefly, we used a combined reverse vaccinology and subtractive genomics approach. As a result, we identified 12 common putative antigenic proteins as vaccine targets and three common drug targets against Mycobacterium leprae and Mycobacterium lepromatosis. Furthermore, the docking analysis using 28 natural compounds with three drug targets was done. The bis-naphthoquinone compound Diospyrin (CID 308140) obtained from indigenous plant Diospyros spp. showed the most favored binding affinity against predicted drug targets, which can be a candidate therapeutic target in the future against leprosy.


The management of personal protective equipment during the COVID-19 pandemic: The case of the province of Quebec.
Like other Canadian provinces, Quebec managed shortages of Personal Protective Equipment (PPE) in the early weeks of the COVID-19 pandemic. Two years later, with hindsight, what lessons can we learn from this logistics crisis? It is precisely to better understand the decisions made by the supply chain players in the province of Quebec that this paper was written. To fully understand the events, this paper first describes the Quebec healthcare system. Then it retraces the series of events and actions during the first wave, at a time when it was most challenging to procure PPE. It also specifies the main characteristics of the supply chain in the Quebec healthcare sector. Finally, it analyzed these data to come up with recommendations to help public decision makers adopt better supply chain management practices.


Fertility Regulation in an Economic Crisis.
Substantial international aid is spent reducing the cost of contraception in developing countries, as part of a larger effort to reduce global fertility and increase investment per child worldwide. The importance for fertility behaviors of keeping contraceptive prices low, however, remains unclear. Targeting of subsidies and insufficient price variation have hindered prior attempts to estimate the effect of monetary and non-monetary contraceptive costs on fertility behavior. Using longitudinal survey data from the Indonesia Family Life Survey, we exploit dramatic variation in prices and incomes that was induced by the economic crisis in the late 1990s to pin down the effect of contraceptive availability and costs as well as household resources on contraceptive use and method choice. The results are unambiguous: monetary costs of contraceptives and levels of family economic resources have a very small (and well-determined) impact on contraceptive use and choice of method.


Comparison of demographics, treatment patterns, health care utilization, and costs among elderly patients with extensive-stage small cell and metastatic non-small cell lung cancers.
Limited data exist regarding real-world treatment patterns, resource utilization, and costs of extensive-stage small cell lung cancer (esSCLC) among elderly patients in the United States. While abundant data are available on treatment patterns in metastatic non-small cell lung cancer (mNSCLC), to our knowledge no data exist comparing costs and resource use between patients with esSCLC or mNSCLC. We retrospectively analyzed administrative claims data (2000-2008) of patients aged ≥65 years from the linked Surveillance, Epidemiology and End Results (SEER)-Medicare database. Patients were selected on the basis of having newly diagnosed esSCLC (n=5,855) or mNSCLC (n=24,090) during 1/1/2000-12/31/2005, and were required to have received cancer-directed therapy. Survival and other measures were compared between esSCLC and mNSCLC patients using Kaplan-Meier log-rank and univariate chi-square and t-tests. Study measures were followed from first diagnosis date of either esSCLC or mNSCLC until the earlier of death or end of the database. Survival between the cohorts did not differ significantly: mean of 10.4 months for esSCLC patients versus 11.1 months for mNSCLC; median survival was 7.4 months versus 5.9 months. A higher percentage of mNSCLC patients (vs. esSCLC) received radiation therapy (75.6% vs. 65.4%; P < 0.001) and surgery (13.6% vs. 7.8%; P < 0.001) during the metastatic disease period. Conversely, a higher percentage of esSCLC patients than mNSCLC patients received chemotherapy (85.5% vs. 60.3%; P < 0.001), red blood-cell transfusion (20.7% vs. 10.9%; P < 0.001), platelet transfusion (5.6% vs. 1.8%; P < 0.001), and growth-factor support (59.0% vs. 39.5%; P < 0.001). esSCLC patients incurred higher lifetime disease-related costs ($44,167 vs. $37,932; P < 0.001) and all-cause costs ($70,549 vs. $67,176; P < 0.001) than mNSCLC patients. Lifetime total and disease-related costs per patient were high. Increased use of chemotherapy, supportive care therapies (including growth factors), and disease-related hospitalizations were observed in esSCLC patients as compared with mNSCLC patients. Disease-related and all-cause costs for esSCLC also exceeded those of mNSCLC, except for hospice and skilled nursing services. Survival and per-patient costs for both groups underscore the unmet medical need for more effective therapies in patients with esSCLC or mNSCLC.


Metabolic heat production and evaporation of poultry.
Accurate measurements of gas exchange between an animal and its environment is critical in determining metabolic heat production and respiratory functions of broilers. Information on non-invasive methods to measure gas exchange of broiler chicks and chickens under uncontrolled environmental conditions is lacking in the literature. The aims of this study were: (1) to develop an indirect calorimetric system including a hood that allows gas exchange for chickens, (2) to measure gas exchange and respiratory functions (respiration rate, ventilation rate, and tidal volume) of broiler chickens weighing greater than 250 g, and (3) to calculate heat production and respiratory evaporation of the birds based on measured gas and vapor exchanges. We conducted two trials. The first trial involved 6 broiler chicks evaluated for 6 days in 6 different schedules (6 × 6 Latin square). The chicks were kept inside a heat exchanger with a continuous air flow of 150 mL min-1. The second trial involved 12 birds evaluated for 12 days in 12 different schedules (12 × 12 Latin square). Metabolic heat production and evaporation were influenced by live weight of chicks, varying between evaluation days (P < 0.05). The respiratory functions (tidal volume, ventilation rate, and respiratory rate) varied between days, and were strongly influenced by live weight of the broilers (P < 0.05).


Qualifying label components for effective biosensing using advanced high-throughput SEIRA methodology.
The need for technological progress in bio-diagnostic assays of high complexity requires both fundamental research and constructing efforts on nano-scaled assay recognition elements that can provide unique selectivity and design-enhanced sensitivity features. Nanoparticle induced sensitivity enhancement and its application related to multiplexed capability Surface-Enhanced InfraRed Absorption (SEIRA) assay formats are well suitable for these purposes. The potential of diverse fluorophore-antibody conjugates, being chemisorbed onto low-cost gold nanoparticulate SEIRA substrates, has been explored with respect to their spectral discriminability. These novel biolabels deliver molecular SEIRA fingerprints that have been successfully analyzed by both uni- and multivariate analyzing tools, to discriminate their multiplexing capabilities. We show that this robust spectral encoding via SEIRA fingerprints opens up new opportunities for a fast, reliable and multiplexed high-end screening in biodiagnostics.


Efficacy of a novel, protein-based pneumococcal vaccine against nasopharyngeal carriage of Streptococcus pneumoniae in infants: A phase 2, randomized, controlled, observer-blind study.
Conserved pneumococcal proteins are potential candidates for inclusion in vaccines against pneumococcal diseases. In the first part of a two-part study, an investigational vaccine (PHiD-CV/dPly/PhtD-30) containing 10 pneumococcal serotype-specific polysaccharide conjugates (10VT) combined with pneumolysin toxoid and pneumococcal histidine triad protein D (30μg each) was well tolerated by Gambian children. Part two, presented here, assessed the efficacy of two PHiD-CV/dPly/PhtD formulations against pneumococcal nasopharyngeal carriage (NPC) prevalence in infants. In this phase 2, randomized, controlled, observer-blind trial, healthy infants aged 8-10weeks, recruited from a peri-urban health center, were randomized (1:1:1:1:1:1) into six groups. Four groups received PHiD-CV/dPly/PhtD (10 or 30μg of each protein), PHiD-CV, or 13-valent pneumococcal conjugate vaccine at ages 2-3-4months (3+0 infant schedule) and two groups PHiD-CV/dPly/PhtD-30 or PHiD-CV at 2-4-9months (2+1 infant schedule). The primary objective was impact on non-10VT NPC at ages 5-9-12months. Secondary objectives included confirmatory analysis of protein dose superiority and safety/reactogenicity. Impact on pneumococcal NPC acquisition, bacterial load, and ply and phtD gene sequencing were explored. 1200 infants were enrolled between June 2011 and May 2012. Prevalences of pneumococcal (60-67%) and non-10VT (55-61%) NPC were high at baseline. Across all post-vaccination time points, efficacy of PHiD-CV/dPly/PhtD-10 and PHiD-CV/dPly/PhtD-30 against non-10VT NPC (3+0 schedule) was 1.1% (95% CI -21.5, 19.5) and 2.1% (-20.3, 20.3), respectively; efficacy of PHiD-CV/dPly/PhtD-30 (2+1 schedule) was 0.5% (-22.1, 18.9) versus PHiD-CV. No differences were observed in pneumococcal NPC acquisition, clearance, or bacterial load. Both protein-based vaccines elicited immune responses to pneumococcal proteins. In this high carriage prevalence setting, inclusion of pneumococcal proteins in the PHiD-CV/dPly/PhtD investigational vaccine had no impact on pneumococcal NPC in infants, regardless of protein dose or schedule. Future evaluations will assess its impact against pneumococcal disease endpoints. PATH, GlaxoSmithKline Biologicals SA. ClinicalTrials.gov identifier NCT01262872.


Halo and subsurface scattering in the transparent coating on top of a diffusing material.
Strongly scattering supports coated with thick transparent medium display a bright halo with a characteristic ring shape when illuminated in one point by a thin pencil of light. The halo, whose size is related to the coating thickness, is due to the Fresnel internal reflections of the light scattered by the diffusing support at the coating-air interface. The angular distribution of the reflected light strongly varies over the halo according to the distance from the point initially illuminated, a fact that cannot be observed when a large area of the surface is illuminated as in usual reflectance and bidirectional reflectance distribution function measurements. By considering a Lambertian background and a transparent layer on top of it, both of them being possibly absorbing, we develop a bidirectional subsurface scattering reflectance distribution function model, based on analytical equations and matrix numerical computation, which enables a detailed description of the spatial and angular distribution of the scattered light including the multiple reflections between the background and the coating-air interface. Some applications in which this subsurface scattering phenomenon can be an issue are addressed, such as the reflectance measurement, which can be undervalued when the geometry is not adapted to the coating thickness, or the impact of the phenomenon on heterogeneously colored surfaces such as coated or laminated halftone prints.


Reproductive health and desire for children among young female sex workers in Bangladesh brothels.
Globally, reproductive health research among female sex workers (FSWs) often focuses on pregnancy prevention, but many women who sell sex aspire to have children in the future. In Bangladesh, where early marriage and parenthood is the norm, we examine reproductive histories and childbearing desires of young women who sell sex in brothels. We interviewed 1061 FSWs aged 18 to 24 in eight brothels in three Bangladesh divisions. Interviewers elicited information on sociodemographic characteristics, contraceptive use, pregnancy history, and childbearing desire. Bivariate and multivariate analyses were conducted to examine correlates of wanting to have a child within 24 months.


Involvement of apoptosis in 4-vinylcyclohexene diepoxide-induced ovotoxicity in rats.
Previous studies have determined that 4-vinylcyclohexene diepoxide (VCD) causes specific destruction of oocytes contained in small pre-antral (primordial and primary) ovarian follicles of Fischer 344 rats following 30 days of daily dosing with VCD. The purposes of this study were to identify the type of VCD-induced cell death occurring in small pre-antral follicles and to determine the earliest time following the onset of dosing when evidence of follicular destruction could first be detected. A significant decrease in the number of oocytes contained in small pre-antral follicles in ovaries of rats after 15 days of daily dosing (ip) with VCD (80 mg/kg) had been observed in preliminary experiments. Therefore, a study was conducted to determine the time of the onset of this follicular destruction by examination of follicular DNA integrity. Female Fischer 344 rats were dosed daily (80 mg/kg, i.p.) for 6, 8, 10, 12, or 14 days, and ovaries were removed 1, 4, or 24 hr after the final dose. Small pre-antral follicles (25-100 microns) were isolated by gentle dissociation of ovaries with collagenase, and follicles were sorted with micropipets. Genomic DNA was isolated from follicles and radiolabeled with [32P]dideoxy ATP, and the degree of fragmentation quantified by agarose gel electrophoresis and autoradiography. Degradation of DNA was evaluated by 32P content in low-molecular-weight fragments ( < 4 kilobase pairs). Degradation of DNA was not observed in follicles collected 24 hr after the final dose on any day. However, a random pattern of DNA degradation was observed, and was significantly greater (p < 0.05) compared with controls, when follicles were collected 4 hr following VCD administration on Days 10 and 12, but not on Days 6 or 8, of dosing. Although not significant, there was also evidence of DNA degradation in dosed animals on Day 14. Histological evaluation of small pre-antral follicles in ovarian sections during the early stages of VCD-induced DNA degradation (Day 10; 4 hr) demonstrated margination of chromatin along the nuclear membrane in oocytes and disruptions in focal contact between granulosa cells and oocytes, both features indicative of apoptosis. Furthermore, there was no sign of ruptured membranes in granulosa cells or oocytes or of an inflammatory response, characteristics of necrosis (pathological cell death). Whereas biochemical and morphological evidence of follicular destruction was seen 4 hr after dosing on Day 10, numbers of oocyte-containing primordial and primary follicles in VCD-treated animals were not different from controls at that time. These results demonstrate that the initial evidence of impending destruction of small pre-antral follicles is first consistently visualized following 10 days of daily dosing with VCD, although a measurable reduction in oocyte numbers has not yet occurred. Despite the fact that internucleosomal cleavage of genomic DNA was not observed, morphological evaluations support that granulosa cells and oocytes in primordial and primary follicles are destroyed via the induction of apoptosis.


In situ measurement of spectral changes in the anterior eye following application of ultraviolet-absorbing compounds.
The ocular structures are very sensitive to damage from ultraviolet (UV) radiation, exposure is linked to corneal and conjunctival damage, cataract formation and may also be implicated in the aetiology of age-related macular degeneration. These structures are usually protected by wearing suitable eyeglasses and goggles. An alternative to conventional eyeglasses/goggles is the concept of "liquid sunglasses" which involve the topical application of eye drops that are designed to block harmful UV radiation reaching the sensitive ocular surfaces. The evaluation of such compounds directly applied to the eye surface requires in situ measurements to compare the efficacy of different formulations. A novel ocular spectrometer system has been used to evaluate changes in the transmission of ultraviolet (UV) radiation through the anterior eye following topical application of candidate UV-absorbing formulations. The key feature of the system is the ability to propagate a beam of light tangentially through the anterior eye using a compact, hand-held lens assembly incorporating UV-transmitting optical fibres. A range of formulations containing UV-absorbing compounds were topically applied to ex vivo rabbit eyes. Significant increases in the absorption of the UV spectrum were detected in seven of the eight formulations studied, demonstrating the potential of this measurement technique in the evaluation of formulations developed as potential topical ocular sunscreens.