In-group biases and oculomotor responses: beyond simple approach motivation.
An in-group bias describes an individual's bias towards a group that they belong to. Previous studies suggest that in-group bias facilitates approach motor responses, but disrupts avoidance ones. Such motor biases are shown to be more robust when the out-group is threatening. We investigated whether, under controlled visual familiarity and complexity, in-group biases still promote pro-saccade and hinder anti-saccades oculomotor responses. Participants first learned to associate an in-group or out-group label with an arbitrary shape. They were then instructed to listen to the group-relevant auditory cue (name of own and a rival university) followed by one of the shapes. Half of the participants were instructed to look towards the visual target if it matched the preceding group-relevant auditory cue and to look away from it if it did not match. The other half of the participants received reversed instructions. This design allowed us to orthogonally manipulate the effect of in-group bias and cognitive control demand on oculomotor responses. Both pro- and anti-saccades were faster and more accurate following the in-group auditory cue. Independently, pro-saccades were performed better than anti-saccades, and match judgements were faster and more accurate than non-match judgements. Our findings indicate that under higher cognitive control demands individuals' oculomotor responses improved following the motivationally salient cue (in-group). Our findings have important implications for learning and cognitive control in a social context. As we included rival groups, our results might to some extent reflect the effects of out-group threat. Future studies could extend our findings using non-threatening out-groups instead.


Glaucoma in aphakia and pseudophakia in the Chennai Glaucoma Study.
To determine the prevalence of glaucoma among aphakes and pseudophakes in a rural population of southern India. 3924 subjects aged 40 years or above underwent complete ophthalmic examination. Glaucoma in aphakia/pseudophakia was diagnosed using International Society of Geographical and Epidemiological Ophthalmology criteria in aphakic/pseudophakic people. 54 subjects (37 aphakes, 17 pseudophakes) (1.38% of 3924 subjects, 11.2% of 482 aphakes/pseudophakes) had glaucoma in aphakia/pseudophakia. Aphakia, age, intraocular pressure (IOP), pseudoexfoliation, and peripheral anterior synechiae greater than or equal to 180 degrees of the angle were risk factors for glaucoma on univariate analysis. On multivariate analysis, IOP and aphakia were independent risk factors for glaucoma. 39 people (72.22%) with glaucoma had normal IOP at presentation. None of the people with glaucoma were aware of the disease. Blindness in one or both eyes was seen in 12 subjects (10 unilateral and two bilateral)-that is, 22.22% of people with glaucoma in aphakia/pseudophakia. Glaucoma is an important cause of ocular morbidity among aphakes and pseudophakes in this rural population of south India. This glaucoma, responsible for unilateral or bilateral blindness in 22.2% of those affected, was entirely undetected in this study population.


Advocacy of home telehealth care among consumers with chronic conditions.
To describe use of home telehealth care as an alternative for chronic disease management from users' perspective. As the population ages, telehealth is increasingly being used to tackle problems related to the fast growing ageing population. Home telehealth care therefore poses challenges and offers opportunities for patients and healthcare providers. A qualitative approach was adopted with a purposeful sample of 20 patients residing in Taiwan. Patients who had received the service for three months and were willing to share their experiences were recruited. Data were collected by face-to-face interviews with semi-structured interview guideline (n = 8) and a focus group discussion (n = 12) in 2010. A qualitative content analysis was used. Four key themes were identified: perceived support and security, enhanced disease self-management, concerned with using the devices and worries about the cost by patients. Most users favoured using the service to control their chronic conditions because of its convenience and accessibility, and their condition could be measured daily to enhance their sense of security. Users could determine and understand changes in their condition and improve medical regimen compliance, and they were empowered to revise their lifestyles for better disease self-management. However, users were concerned about the utility of the service, because they were unfamiliar with the operating procedures and doubted its quality. As the service is still in stage of pilot testing, users worried about possible cost and reimbursement policy changes in the future. Most users perceived telehealth care was a convenient and useful model for healthcare-delivery. It increased the availability of health care and improved the self-care ability of patients. To advocate for home telehealth care, nurses must play an active role in providing consumers with proper training and support for any problems when adopting the system to foster patients' willingness to use this service.


Biogenic habitat shifts under long-term ocean acidification show nonlinear community responses and unbalanced functions of associated invertebrates.
Experiments have shown that increasing dissolved CO2 concentrations (i.e. Ocean Acidification, OA) in marine ecosystems may act as nutrient for primary producers (e.g. fleshy algae) or a stressor for calcifying species (e.g., coralline algae, corals, molluscs). For the first time, rapid habitat dominance shifts and altered competitive replacement from a reef-forming to a non-reef-forming biogenic habitat were documented over one-year exposure to low pH/high CO2 through a transplant experiment off Vulcano Island CO2 seeps (NE Sicily, Italy). Ocean acidification decreased vermetid reefs complexity via a reduction in the reef-building species density, boosted canopy macroalgae and led to changes in composition, structure and functional diversity of the associated benthic assemblages. OA effects on invertebrate richness and abundance were nonlinear, being maximal at intermediate complexity levels of vermetid reefs and canopy forming algae. Abundance of higher order consumers (e.g. carnivores, suspension feeders) decreased under elevated CO2 levels. Herbivores were non-linearly related to OA conditions, with increasing competitive release only of minor intertidal grazers (e.g. amphipods) under elevated CO2 levels. Our results support the dual role of CO2 (as a stressor and as a resource) in disrupting the state of rocky shore communities, and raise specific concerns about the future of intertidal reef ecosystem under increasing CO2 emissions. We contribute to inform predictions of the complex and nonlinear community effects of OA on biogenic habitats, but at the same time encourage the use of multiple natural CO2 gradients in providing quantitative data on changing community responses to long-term CO2 exposure.


Validity of New Ballard Score until 7th day of postnatal life in moderately preterm neonates.
The New Ballard Score (NBS) has been evaluated only until 96 h of age. We studied the validity and reliability of NBS for gestational age (GA) assessment on days 1, 5 and 7 of postnatal age (PNA). This prospective, analytical study was conducted in a level III neonatal unit. Neonates born at a GA of 29-35 weeks (based on accurate last menstrual period (LMP)) were eligible. Encephalopathy, malformations, and unstable vitals were exclusion criteria. LMP-based GA was the gold standard. NBS was assessed within 24 h of birth by one rater, and two raters assessed NBS on days 5 and 7. All were blinded to LMP and one anothers' ratings. Recruitment continued until >100 subjects were enrolled with > or =25 in each LMP-based GA group: 29-30 weeks, 31-32 weeks, 33-34 weeks and 35 weeks. Correlation of GA assessed on day 7 with gold standard. 129 neonates were studied. NBS-based GA on days 5 or 7 did not differ from the gold standard GA by more than 2 weeks in any subject. On day 7, NBS overestimated GA in 26.7% and underestimated GA in 19.8% cases; all discrepancies were < or =2 wks. Compared to gold standard GA, the intra-class correlations (ICCs) of the gold standard GA and the NBS-based GA of the day 1 rater, day 5 rater and day 7 rater were 0.94, 0.94 and 0.92, respectively. ICCs for inter-rater reliability on day 5 and day 7 were 0.97 and 0.96, respectively. Compared to the day 1 rater's raw NBS, the ICCs of day 5 and day 7 raters' total scores were 0.98 and 0.97, respectively; of day 5 and day 7 raters' neurological scores were 0.98 and 0.97; and of day 5 and day 7 raters' physical scores were 0.92 and 0.88. All ICCs mentioned above had p values <0.001. NBS is a valid and reliable clinical tool for GA assessment until day 7. It slightly overestimates the GA with increasing PNA. Neurological signs are more reliable than physical ones.


Effects of histone deacetylase inhibitors on estradiol-induced proliferation and hyperplasia formation in the mouse uterus.
It is suggested that estrogen hormones recruit mechanisms controlling histone acetylation to bring about their effects in the uterus. However, it is not known how the level of histone acetylation affects estrogen-dependent processes in the uterus, especially proliferation and morphogenetic changes. Therefore, this study examined the effects of histone deacetylase blockers, trichostatin A and sodium butyrate, on proliferative and morphogenetic reactions in the uterus under long-term estrogen treatment. Ovari-ectomized mice were treated with estradiol dipropionate (4 microg per 100 g; s.c., once a week) or vehicle and trichostatin A (0.008 mg per 100 g; s.c., once a day) or sodium butyrate (1% in drinking water), or with no additional treatments for a month. In animals treated with estradiol and trichostatin A or sodium butyrate, uterine mass was increased, and abnormal uterine glands and atypical endometrial hyperplasia were found more often. Both histone deacetylase inhibitors produced an increase in the numbers of mitotic and bromodeoxyuridine-labelled cells in luminal and glandular epithelia, in stromal and myometrial cells. Levels of estrogen receptor-alpha and progesterone receptors in uterine epithelia, stromal and myometrial cells were decreased in mice treated with estradiol and trichostatin A or sodium butyrate. Expression of beta-catenin in luminal and glandular epithelia was attenuated in mice treated with estradiol with trichostatin A or sodium butyrate. Both histone deacetylase inhibitors have similar unilateral effects; however the action of trichostatin A was more expressed than that of sodium butyrate. Thus, histone deacetylase inhibitors exert proliferative and morphogenetic effects of estradiol. The effects of trichostatin A and sodium butyrate are associated with changes in expression of estrogen receptor-alpha, progesterone receptors and beta-catenin in the uterus.


A synthesis of qualitative research exploring the barriers to staying in work with chronic musculoskeletal pain.
Qualitative research can help to advance our understanding, management and prevention of work disability. Our aim was to integrate qualitative research findings in order to increase our understanding of barriers to stay in work with chronic pain. We searched five electronic bibliographic databases until September 2012, supplemented by citation tracking and hand-searching. We used meta-ethnography to synthesis our findings. Central to meta-ethnography is identifying "concepts" and developing a conceptual model. Concepts were compared and organised into categories. The following categories can have an impact on the decision to remain in work: struggling to affirm myself as a good worker; balancing life and work in the face of unpredictable symptoms; my work colleagues don't believe me; the system does not facilitate return to work; the battle for legitimacy. Our innovation is to present an internationally relevant model based on a conceptual synthesis. This model highlights the adversarial work experience of people with chronic. The papers span 15 years of qualitative research. A significant finding is that these themes continue to pervade the current work environment for those in pain, and this has clear implications for education, social care and policy. People with chronic pain face an adversarial struggle to maintain their credibility at work. Strategies to maintain personal credibility can have an adverse effect on working lives. Changes at a systems level are needed to facilitate continuance and return to work. Cultural changes in the way that we view people with pain would help to keep people in work.


Aspirin does not prevent the attenuation of myocardial stunning by the ACE inhibitor ramiprilat.
The attenuation of myocardial stunning by the ACE inhibitor ramiprilat is prevented by cyclooxygenase inhibition with indomethacin. In the clinical setting of ischemia/reperfusion however, the cyclooxygenase inhibitor aspirin is widely used to prevent platelet aggregation. The present study therefore investigated whether aspirin in dosages sufficient to inhibit platelet aggregation interferes with the attenuation of myocardial stunning by ramiprilat. Fifteen dogs received either 1 mg/(kg.day) (group I, n = 7) or 10 mg/(kg.day) (group II, n = 8) aspirin orally for 1 week. Both dosages of aspirin inhibited ADP-induced platelet aggregation. The dogs were then anesthetized thoracotomized and subjected to 15 min LCx-occlusion and 4 h reperfusion. Before LCx-occlusion, groups I and II received ramiprilat (20 micrograms/kg, i.v.). Systemic hemodynamics, posterior myocardial blood flow (PMBF, colored microspheres) and wall thickening (PWT, sonomicrometry) of these groups were measured and data compared to placebo-controls (group III, n = 11) and dogs receiving only ramiprilat before LCx-occlusion (group IV, n = 11). Four dogs received 10 mg/(kg.day) aspirin without ramiprilat (group V). Mean aortic pressure was kept constant by an intra-aortic balloon, and heart rate did not change. PMBF was not different between the five groups. Under control conditions and during myocardial ischemia PWT was also not different. At 4 h reperfusion PWT was still depressed in group III (-5 +/- 20% of control) and group V (-23 +/- 6%) whereas PWT recovered to the same extent in groups I (46 +/- 23%), II (50 +/- 15%) and IV (58 +/- 18%) (all P < 0.05 v groups III and V). The attenuation of myocardial stunning by the ACE inhibitor ramiprilat is not prevented by aspirin in dosages which are nevertheless sufficient to inhibit platelet aggregation.


5 flourouracil-induced apical ballooning syndrome: a case report.
The apical ballooning syndrome (ABS) is a recently described stress-mediated acute cardiac syndrome characterized by transient wall-motion abnormalities involving the apex and midventricle with hyperkinesis of the basal left ventricular (LV) segments without obstructive epicardial coronary disease. Cardiotoxicity is not an uncommon adverse effect of chemotherapeutic agents. However, there are no reports of ABS secondary to chemotherapeutic agents. We describe the case of a woman who developed the syndrome after chemotherapy for metastatic cancer. A 79-year-old woman presented with typical ischemic chest pain, elevated cardiac enzymes with significant ST-segment abnormalities on her electrocardiogram. She underwent recent chemotherapy with fluorouracil for metastatic colorectal cancer. Echocardiography revealed a wall-motion abnormality involving the apical and periapical segments which appeared akinetic. Coronary angiography revealed no obstructive coronary lesions. The patient was stabilized with medical therapy. Four weeks later she remained completely asymptomatic. Echocardiogram revealed a normal ejection fraction and a resolution of the apical akinesis. Pathogenetic mechanisms of cardiac complications in cancer patients undergoing chemotherapy include coronary vasospasm, endothelial damage and consequent thrombus formation. In our patient, both supraphysiologic levels of plasma catecholamines and stress related neuropeptides caused by cancer diagnosis as well as chemotherapy may have contributed the development of ABS.


Analysis of Ocular Firework-Related Injuries and Common Eye Traumata: a 5-year Clinical Study.
Background A comparative study of eye injuries related to fireworks or acts of violence around New Year's Eve and the Swiss National Day on August 1st. The two groups were compared with respect to the overall numbers of eye accidents within the period of review. Patients and Methods Retrospective analysis of emergency consultations at the Department of Ophthalmology, University Hospital Zurich with eye accidents around the Swiss National Day on August 1st and New Year's Eve over the last 5 years. Two subgroups were formed: (1) Firework-related eye traumata, (2) Eye injuries due to acts of violence. The groups were analysed by age, gender, active participant or bystander, eye involved, severity of trauma (from clinical findings), surgical interventions, time of follow-up and visits, visual acuity and outcome. Results The study included 97 patients (100 eyes) with 74 male (76 %) and 23 female (24 %) victims. After filtering out 67 common traumata cases (all unilateral), 17 patients (18 eyes) with firework-related injuries and 13 patients (15 eyes) with damage due to an act of violence remained. Firework injuries accounted for 18 % of cases (65 % men); eye injuries caused by an act of violence accounted for 15 % of cases (92 % men). In the fireworks group, women were significantly older than men (mean age men 32 ± 14 years versus women 38 ± 16 years, p = 0.002). 65 % of cases were bystanders. The two subgroups contained 30 patients (33 eyes) with 22 left eyes (67 %, p < 0.001). The anterior segment was most frequently involved (79 %), significantly more often than the posterior part of the eye (p < 0.001). The posterior segment was injured more often in the fireworks group (28 %), than in the violence group (13 %). 87 % of the victims in the group of common traumata were mild trauma, 10 % moderate and 3 % severe. In the fireworks group the distribution was 53 % mild, 12 % moderate and 35 % severe, in the violence group 46 %, 23 % and 31 % respectively. Severe trauma cases occurred significantly more often around the New Year, with 36 % versus August 1st with 18 % (p = 0.0028). They are more often related to firework injuries than caused by violence (40 % versus 33 %). In summary, 7 victims (23 %) needed 8 interventions, with 3 having additional issues with their intraocular pressure. Full final visual acuity was noted in 88 %, which was alike in the two subgroups. On the contrary common traumata happened significantly more often around August 1st (66 %) than New Year (34 %; p < 0.001). Conclusions The number of injured eyes in the two subgroups of fireworks and violence was twice as high around New Year's Eve than around the Swiss National Day. If the two events were combined, eye injuries were caused by an act of violence in 45 % and by fireworks in 55 % of cases. Injuries from fireworks can be decreased by professional fireworks. Both types of accident can be decreased by public awareness.


Effects of training on indices of iron status of young female cross-country runners.
Hematological status and selected indices of iron status were assessed in a group of eight high-school female cross-country runners at weeks 0, 1, 2, 4, and 8 of the competitive season and at 1, 5, and 11 wk after the season. Over the season, a training effect occurred, as was shown by a significant decrease (12.8%) in step-test heart rate and a significant increase (7.0%) in VO2max (ml X min-1 X kg-1). The runners experienced "sports anemia" in that their hemoglobin (Hb) concentration and packed red blood cell volume (PCV) declined significantly during the first week of training (8.8 and 8.3%, respectively), whereas changes did not occur in a comparison group of 11 nonrunners who were followed for the first 3 wk of the season. Between weeks 1 and 8 of the season, the runners' Hb and PCV values gradually returned toward preseason values while their serum iron (SeFe) and percent transferrin saturation (% Sat) showed steady, but nonsignificant, declines, and their total iron-binding capacity (TIBC) rose by 9.4% and continued to rise through the first week after the season. Concomitantly, the runners' free erythrocyte porphyrin (FEP) concentrations rose by 15%. By the end of the detraining period, all indices of iron status had returned to initial values except for TIBC, which still was significantly higher than preseason values. With the exception of TIBC, no significant changes occurred in the iron-status indices of the comparison group during the 2 wk they were followed. The results suggest that in young women recovery from sports anemia may impose a demand on body iron reserves and that it would be prudent to assess iron status as well as hematological status in such women before and during aerobic training.


Genomic diversity of 2010 Haitian cholera outbreak strains.
The millions of deaths from cholera during the past 200 y, coupled with the morbidity and mortality of cholera in Haiti since October 2010, are grim reminders that Vibrio cholerae, the etiologic agent of cholera, remains a scourge. We report the isolation of both V. cholerae O1 and non-O1/O139 early in the Haiti cholera epidemic from samples collected from victims in 18 towns across eight Arrondissements of Haiti. The results showed two distinct populations of V. cholerae coexisted in Haiti early in the epidemic. As non-O1/O139 V. cholerae was the sole pathogen isolated from 21% of the clinical specimens, its role in this epidemic, either alone or in concert with V. cholerae O1, cannot be dismissed. A genomic approach was used to examine similarities and differences among the Haitian V. cholerae O1 and V. cholerae non-O1/O139 strains. A total of 47 V. cholerae O1 and 29 V. cholerae non-O1/O139 isolates from patients and the environment were sequenced. Comparative genome analyses of the 76 genomes and eight reference strains of V. cholerae isolated in concurrent epidemics outside Haiti and 27 V. cholerae genomes available in the public database demonstrated substantial diversity of V. cholerae and ongoing flux within its genome.


Structure of the amino terminus of a gap junction protein.
Charged amino acid residues in the amino terminus of gap junction forming proteins (connexins) form part, if not all, of the transjunctional voltage sensor of gap junction channels and play a fundamental role in ion permeation. Results from studies of the voltage dependence of N-terminal mutants predict that residues 1-10 of Group I connexins lie within the channel pore and that the N-terminus forms the channel vestibule by the creation of a turn initiated by the conserved G12 residue. Here we report that intercellular channels containing mutations of G12 in Cx32 to residues that are likely to interfere with flexibility of this locus (G12S, G12Y, and G12V) do not express junctional currents, whereas a connexin containing a proline residue at G12 (Cx32G12P), which is expected to maintain a structure similar to that of the G12 locus, forms nearly wild-type channels. We have solved the structure of an N-terminal peptide of Cx26 (MDWGTLQSILGGVNK) using 1H 2D NMR. The peptide contains two structured domains connected by a flexible hinge (domain-hinge-domain motif) that would allow the placement of the amino terminus within the channel pore. Residues 1-10 adopt a helical conformation and line the channel entrance while residues 12-15 form an open turn. Overall, there is good agreement between the structural and dynamic features of the N-terminal peptide provided by NMR and the functional studies of the voltage dependence of channels formed by wild-type and N-terminal mutations.


Evaluation of Effect of Zirconia Surface Treatment, Using Plasma of Argon and Silane, on the Shear Bond Strength of Two Composite Resin Cements.
Yttria stabilised tetragonal zirconia opens new vistas for all ceramic restoration by the mechanism of transformation toughening, making it much stronger compared to all other ceramic materials. Currently, it is the most recent core material for all ceramic fixed partial dentures due to its ability to withstand high simulated masticatory loads. Problems which have been reported with zirconia restorations involve the core cement interface leading to loss of retention of the prosthesis. Different reasons which have been reported for the same include the lack of adhesion between zirconia and commonly used cements due to absence of silica phase which makes zirconia not etchable. In addition, the hydrophobic nature of zirconia causes low wettability of zirconia surface by the adhesive cements which are commonly used. The purpose of this in vitro study was to compare and evaluate the effect of two pre-treatments of zirconia, using plasma of argon and silane, on the shear bond strength values of two composite resin cements to zirconia and to evaluate the failure pattern of the debonded areas using stereomicroscopic analysis. Sixty zirconia discs (10 mm×2 mm) were randomly divided into three groups (n=20), following surface treatment, with airborne particle abrasion, using 110 µm Al2O3: Group I (control), Group II (plasma of argon cleaning), and Group III (application of silane primer). Each group had two subgroups based on the type of resin cement used for bonding: subgroup A; Rely X Ultimate (3M ESPE) and subgroup B; Panavia F (Kuraray). In subgroup A, Rely X universal silane primer and in subgroup B Clearfil ceramic primer was used. Shear bond strengths were determined after water storage for one day and thermocycling for 5000 cycles. Data (megapascal) were analyzed using ANOVA and Bonferroni test. Specimens were subjected to stereomicroscopic analysis, for evaluation of failure pattern. Group III produced the highest shear bond strength followed by Group II and Group I. Subgroup A showed higher shear bond strength than Subgroup B. Stereomicroscopic analysis showed cohesive failure in Group III, while in Group I adhesive failure was seen. Silane primer application caused maximum increase in shear bond strength due to increased wettability. Argon plasma treatment was less effective in comparison to silane treatment. Air abrasion when used alone resulted in lower bond strength values, thereby making it necessary to use a combination of surface treatments. Rely X Ultimate cement was superior to Panavia F in terms of adhesive bonding to zirconia.


Low-Bias RNA Sequencing of the HIV-2 Genome from Blood Plasma.
Accurate determination of the genetic diversity present in the HIV quasispecies is critical for the development of a preventative vaccine: in particular, little is known about viral genetic diversity for the second type of HIV, HIV-2. A better understanding of HIV-2 biology is relevant to the HIV vaccine field because a substantial proportion of infected people experience long-term viral control, and prior HIV-2 infection has been associated with slower HIV-1 disease progression in coinfected subjects. The majority of traditional and next-generation sequencing methods have relied on target amplification prior to sequencing, introducing biases that may obscure the true signals of diversity in the viral population. Additionally, target enrichment through PCR requires a priori sequence knowledge, which is lacking for HIV-2. Therefore, a target enrichment free method of library preparation would be valuable for the field. We applied an RNA shotgun sequencing (RNA-Seq) method without PCR amplification to cultured viral stocks and patient plasma samples from HIV-2-infected individuals. Libraries generated from total plasma RNA were analyzed with a two-step pipeline: (i) de novo genome assembly, followed by (ii) read remapping. By this approach, whole-genome sequences were generated with a 28× to 67× mean depth of coverage. Assembled reads showed a low level of GC bias, and comparison of the genome diversities at the intrahost level showed low diversity in the accessory gene vpx in all patients. Our study demonstrates that RNA-Seq is a feasible full-genome de novo sequencing method for blood plasma samples collected from HIV-2-infected individuals.IMPORTANCE An accurate picture of viral genetic diversity is critical for the development of a globally effective HIV vaccine. However, sequencing strategies are often complicated by target enrichment prior to sequencing, introducing biases that can distort variant frequencies, which are not easily corrected for in downstream analyses. Additionally, detailed a priori sequence knowledge is needed to inform robust primer design when employing PCR amplification, a factor that is often lacking when working with tropical diseases localized in developing countries. Previous work has demonstrated that direct RNA shotgun sequencing (RNA-Seq) can be used to circumvent these issues for hepatitis C virus (HCV) and norovirus. We applied RNA-Seq to total RNA extracted from HIV-2 blood plasma samples, demonstrating the applicability of this technique to HIV-2 and allowing us to generate a dynamic picture of genetic diversity over the whole genome of HIV-2 in the context of low-bias sequencing.


History of Mosquitoborne Diseases in the United States and Implications for New Pathogens.
The introduction and spread of West Nile virus and the recent introduction of chikungunya and Zika viruses into the Americas have raised concern about the potential for various tropical pathogens to become established in North America. A historical analysis of yellow fever and malaria incidences in the United States suggests that it is not merely a temperate climate that keeps these pathogens from becoming established. Instead, socioeconomic changes are the most likely explanation for why these pathogens essentially disappeared from the United States yet remain a problem in tropical areas. In contrast to these anthroponotic pathogens that require humans in their transmission cycle, zoonotic pathogens are only slightly affected by socioeconomic factors, which is why West Nile virus became established in North America. In light of increasing globalization, we need to be concerned about the introduction of pathogens such as Rift Valley fever, Japanese encephalitis, and Venezuelan equine encephalitis viruses.


Mutations in FAM20C are associated with lethal osteosclerotic bone dysplasia (Raine syndrome), highlighting a crucial molecule in bone development.
The generation and homeostasis of bone tissue throughout development and maturity is controlled by the carefully balanced processes of bone formation and resorption. Disruption of this balance can give rise to a broad range of skeletal pathologies. Lethal osteosclerotic bone dysplasia (or, Raine syndrome) is an autosomal recessive disorder characterized by generalized osteosclerosis with periosteal bone formation and a distinctive facial phenotype. Affected individuals survive only days or weeks. We have identified and defined a chromosome 7 uniparental isodisomy and a 7p telomeric microdeletion in an affected subject. The extent of the deleted region at the 7p telomere was established by genotyping microsatellite markers across the telomeric region. The region is delimited by marker D7S2563 and contains five transcriptional units. Sequence analysis of FAM20C, located within the deleted region, in six additional affected subjects revealed four homozygous mutations and two compound heterozygotes. The identified mutations include four nonsynonymous base changes, all affecting evolutionarily conserved residues, and four splice-site changes that are predicted to have a detrimental effect on splicing. FAM20C is a member of the FAM20 family of secreted proteins, and its mouse orthologue (DMP4) has demonstrated calcium-binding properties; we also show by in situ hybridization its expression profile in mineralizing tissues during development. This study defines the causative role of FAM20C in this lethal osteosclerotic disorder and its crucial role in normal bone development.


Foot pathology in patients with Paget's disease of bone.
The characteristic bone deformities associated with advanced Paget's disease of bone may result in abnormal gait and plantar foot pressures, which contribute to the development of pressure-related skin problems. This study aimed to characterize the foot problems and foot-care needs in this patient group and to investigate the effect of disease distribution on the occurrence of foot pathology. One hundred thirty-four patients with Paget's disease were assessed clinically, and the Foot Structure Index was completed for each patient. Patients completed self-administered questionnaires concerning foot function (Foot Function Index) and quality of life (12-Item Short Form). The results of this study suggest that foot-health and footwear problems are common in patients with Paget's disease. Forty percent of the patients required professional foot care in addition to those already receiving it. The site of pagetic involvement did not affect the occurrence of foot pathology. Further research is required on the impact of the extent of any femoral or tibial deformity on foot symptoms and pathology. Biomechanical studies of the forces and motion in the foot related to different degrees of femoral and tibial deformity might also help determine the impact of deformity on foot pathology.


An Msh2 conditional knockout mouse for studying intestinal cancer and testing anticancer agents.
Mutations in the DNA mismatch repair (MMR) gene MSH2 cause Lynch syndromes I and II and sporadic colorectal cancers. Msh2(null) mice predominantly develop lymphoma and do not accurately recapitulate the colorectal cancer phenotype. We generated and examined mice with a conditional Msh2 disruption (Msh2(LoxP)), permitting tissue-specific gene inactivation. ECMsh2(LoxP/LoxP) mice carried an EIIa-Cre transgene, and VCMsh2(LoxP/LoxP) mice carried a Villin-Cre transgene. We combined the VCMsh2(LoxP) allele with either Msh2(Delta7null) (VCMsh2(LoxP/null)) or Msh2(G674D) mutations (VCMsh2(LoxP/G674D)) to create allelic phase mutants. These mice were given cisplatin or 5-fluorouracil/leucovorin and oxaliplatin (FOLFOX), and their tumors were measured by magnetic resonance imaging. Embryonic fibroblasts from ECMsh2(LoxP/LoxP) mice do not express MSH2 and are MMR deficient. Reverse transcription, polymerase chain reaction, and immunohistochemistry from VCMsh2(LoxP/LoxP) mice demonstrated specific loss of Msh2 messenger RNA and protein from epithelial cells of the intestinal tract. Microsatellite instability was observed in all VCMsh2 strains and limited to the intestinal mucosa. Resulting adenomas and adenocarcinomas had somatic truncation mutations to the adenomatous polyposis coli (Apc) gene. VCMsh2(LoxP/LoxP) mice did not develop lymphoma. Comparison of allelic phase tumors revealed significant differences in multiplicity and size. When treated with cisplatin or FOLFOX, tumor size was reduced in VCMsh2(LoxP/G674D) but not VCMsh2(LoxP/null) tumors. The apoptotic response to FOLFOX was partially sustained in the intestinal mucosa of VCMsh2(LoxP/G674D) animals. Msh2(LoxP/LoxP) mice in combination with appropriate Cre recombinase transgenes have excellent potential for preclinical modeling of Lynch syndrome, MMR-deficient tumors of other tissue types, and use in drug development.


How nervous systems evolve in relation to their embodiment: what we can learn from octopuses and other molluscs.
Cephalopods such as the octopus show the most advanced behavior among invertebrates, which they accomplish with an exceptionally flexible body plan. In this review I propose that the embodied organization approach, developed by roboticists to design efficient autonomous robots, is useful for understanding the evolution and development of the efficient adaptive interaction of animals with their environment, using the octopus as the leading example. The embodied organization approach explains adaptive behavior as emerging from the continuous dynamical and reciprocal physical and informational interactions between four elements: the controller, the mechanical and the sensory systems and the environment. In contrast to hierarchical organization, in embodied organization, self-organization processes can take part in the emergence of the adaptive properties. I first discuss how the embodiment concept explains covariation of body form, nervous system organization, and level of behavioral complexity using the Mollusca as an example. This is an ideal phylum to test such a qualitative correlation between body/brain/behavior, because they show the greatest variations of body plan within a single phylum. In some cases the covariation of nervous system and body structure seems to arise independently of close phylogenetic relationships. Next, I dwell on the octopus as an ideal model to test the embodiment concept within a single biological system. Here, the unusual body morphology of the octopus exposes the uniqueness of the four components comprising the octopus' embodiment. Considering together the results from behavioral, physiological, anatomical, and motor control research suggests that these four elements mutually influence each other. It is this mutual interactions and self-organization which have led to their unique evolution and development to create the unique and highly efficient octopus embodiment.


Comparison of the effects of bupropion and nicotine on locomotor activation and dopamine release in vivo.
Bupropion is an atypical anti-depressant that is approved for smoking cessation. In addition to inhibiting dopamine reuptake, bupropion has been reported to block nicotinic acetylcholine receptors in vitro, and this action might contribute to its efficacy for smoking cessation. In this study we investigated if nicotinic receptor-mediated responses in vivo are decreased in the presence of a behaviorally effective dose of bupropion. In separate experiments we measured locomotor activation and dopamine overflow in the nucleus accumbens core, using in vivo microdialysis in freely moving rats. Bupropion (30 mg/kg i.p.) increased locomotor activity, which remained elevated for up to 2 h. Nicotine (0.4 mg/kg s.c.) also increased locomotor activity but for a shorter duration. When given 20 min after bupropion, hyperlocomotion was significantly enhanced, compared to the response to either nicotine or bupropion alone, consistent with the effects of the two drugs being additive. Systemic administration of bupropion (30 mg/kg i.p.) also elicited a significant increase in dopamine overflow (113+/-16% above basal levels). Nicotine (3 mM; delivered into the nucleus accumbens core via the microdialysis probe) increased dopamine overflow by 126 +/- 35%. Nicotine delivered during the response to bupropion resulted in enhanced dopamine overflow of 294 +/- 50%, also consistent with the actions of the two drugs being additive. This study suggests that behaviorally effective concentrations of bupropion in the rat do not diminish the effects of nicotine by blocking nicotinic receptors.


Structural and functional insights into the mode of action of a universally conserved Obg GTPase.
Obg proteins are a family of P-loop GTPases, conserved from bacteria to human. The Obg protein in Escherichia coli (ObgE) has been implicated in many diverse cellular functions, with proposed molecular roles in two global processes, ribosome assembly and stringent response. Here, using pre-steady state fast kinetics we demonstrate that ObgE is an anti-association factor, which prevents ribosomal subunit association and downstream steps in translation by binding to the 50S subunit. ObgE is a ribosome dependent GTPase; however, upon binding to guanosine tetraphosphate (ppGpp), the global regulator of stringent response, ObgE exhibits an enhanced interaction with the 50S subunit, resulting in increased equilibrium dissociation of the 70S ribosome into subunits. Furthermore, our cryo-electron microscopy (cryo-EM) structure of the 50S·ObgE·GMPPNP complex indicates that the evolutionarily conserved N-terminal domain (NTD) of ObgE is a tRNA structural mimic, with specific interactions with peptidyl-transferase center, displaying a marked resemblance to Class I release factors. These structural data might define ObgE as a specialized translation factor related to stress responses, and provide a framework towards future elucidation of functional interplay between ObgE and ribosome-associated (p)ppGpp regulators. Together with published data, our results suggest that ObgE might act as a checkpoint in final stages of the 50S subunit assembly under normal growth conditions. And more importantly, ObgE, as a (p)ppGpp effector, might also have a regulatory role in the production of the 50S subunit and its participation in translation under certain stressed conditions. Thus, our findings might have uncovered an under-recognized mechanism of translation control by environmental cues.


Tailoring the dependency between rigidity and water uptake of a microfabricated hydrogel with the conformational rigidity of a polymer cross-linker.
Many diverse applications utilize hydrogels as carriers, sensors, and actuators, and these applications rely on the refined control of physical properties of the hydrogel, such as elastic modulus and degree of swelling. Often, hydrogel properties are interdependent; for example, when elastic modulus is increased, degree of swelling is decreased. Controlling these inverse dependencies remains a major barrier for broader hydrogel applications. We hypothesized that polymer cross-linkers with varied chain flexibility would allow us to tune the inverse dependency between the elastic modulus and the degree of swelling of the hydrogels. We examined this hypothesis by using alginate and poly(acrylic acid) (PAA) modified with a controlled number of methacrylic groups as model inflexible and flexible cross-linkers, respectively. Interestingly, the polyacrylamide hydrogel cross-linked by the inflexible alginate methacrylates exhibited less dependency between the degree of swelling and the elastic modulus than the hydrogel cross-linked by flexible PAA methacrylates. This critical role of the cross-linker's inflexibility was related to the difference of the degree of hydrophobic association between polymer cross-linkers, as confirmed with pyrene probes added in pregel solutions. Furthermore, hydrogels cross-linked with alginate methacrylates could tune the projection area of adhered cells by solely altering elastic moduli. In contrast, gels cross-linked with PAA methacrylates failed to modulate the cellular adhesion morphology due to a lower, and smaller, elastic modulus range to be controlled. Overall, the results of this study will significantly advance the controllability of hydrogel properties and greatly enhance the performance of hydrogels in various biological applications.


Protection against Chikungunya virus induced arthralgia following prophylactic treatment with adenovirus vectored interferon (mDEF201).
Recent outbreaks of Chikungunya virus (CHIKV) infection have resulted in millions of cases of disease with significant morbidity. No approved antiviral treatments exist for the prevention or treatment of this viral disease. Infection with CHIKV results in a high rate of symptomatic disease that primarily includes a debilitating arthralgia. To model this cardinal disease manifestation, adult DBA/1J mice were challenged with CHIKV by footpad injection. Viremia and hind limb virus titers increased ∼100-fold while spleen virus increased >1000-fold within 1day post-virus infection (dpi). Footpad swelling was measured over a 10-day period, with peak swelling observed between 6 and 7dpi. Histology of the hind leg at the site of virus challenge showed evidence of myositis and synovitis starting on 5dpi. Cytokine profiling of the hind limb at the site of inoculation revealed a biphasic inflammatory response represented by an increase in IL-6, MCP-1, IFN-γ, MIP-1α, RANTES, and IL-17. To investigate the prophylactic capacity of IFN, mice were treated with mDEF201, an adenovirus-vectored IFN-α. Intranasal administration of a single 10(7)pfu/ml dose of mDEF201 administered 21days to 24h prior to infection, significantly reduced footpad swelling, virus titers in the hind leg and spleen, and several inflammatory cytokines. Efficacy was not observed when treatment was initiated 24h after virus challenge. This arthralgia model of CHIKV recapitulates relevant disease features commonly observed in human disease making it applicable to preclinical testing of therapies that target both viral replication and the associated joint disease.


Optical coherence tomography for rapid tissue screening and directed histological sectioning.
In pathology, histological examination of the "gold standard" to diagnose various diseases. It has contributed significantly toward identifying the abnormalities in tissues and cells, but has inherent drawbacks when used for fast and accurate diagnosis. These limitations include the lack of in vivo observation in real time and sampling errors due to limited number and area coverage of tissue sections. Its diagnostic yield also varies depending on the ability of the physician and the effectiveness of any image guidance technique that may be used for tissue screening during excisional biopsy. In order to overcome these current limitations of histology-based diagnostics, there are significant needs for either complementary or alternative imaging techniques which perform non-destructive, high resolution, and rapid tissue screening. Optical coherence tomography (OCT) is an emerging imaging modality which allows real-time cross-sectional imaging with high resolutions that approach those of histology. OCT could be a very promising technique which has the potential to be used as an adjunct to histological tissue observation when it is not practical to take specimens for histological processing, when large areas of tissue need investigating, or when rapid microscopic imaging is needed. This review will describe the use of OCT as an image guidance tool for fast tissue screening and directed histological tissue sectioning in pathology.


Specificity and mechanism of action of some commonly used protein kinase inhibitors.
The specificities of 28 commercially available compounds reported to be relatively selective inhibitors of particular serine/threonine-specific protein kinases have been examined against a large panel of protein kinases. The compounds KT 5720, Rottlerin and quercetin were found to inhibit many protein kinases, sometimes much more potently than their presumed targets, and conclusions drawn from their use in cell-based experiments are likely to be erroneous. Ro 318220 and related bisindoylmaleimides, as well as H89, HA1077 and Y 27632, were more selective inhibitors, but still inhibited two or more protein kinases with similar potency. LY 294002 was found to inhibit casein kinase-2 with similar potency to phosphoinositide (phosphatidylinositol) 3-kinase. The compounds with the most impressive selectivity profiles were KN62, PD 98059, U0126, PD 184352, rapamycin, wortmannin, SB 203580 and SB 202190. U0126 and PD 184352, like PD 98059, were found to block the mitogen-activated protein kinase (MAPK) cascade in cell-based assays by preventing the activation of MAPK kinase (MKK1), and not by inhibiting MKK1 activity directly. Apart from rapamycin and PD 184352, even the most selective inhibitors affected at least one additional protein kinase. Our results demonstrate that the specificities of protein kinase inhibitors cannot be assessed simply by studying their effect on kinases that are closely related in primary structure. We propose guidelines for the use of protein kinase inhibitors in cell-based assays.


Aqueous Extract of Phyllanthus niruri Leaves Displays In Vitro Antioxidant Activity and Prevents the Elevation of Oxidative Stress in the Kidney of Streptozotocin-Induced Diabetic Male Rats.
P. niruri has been reported to possess antidiabetic and kidney protective effects. In the present study, the phytochemical constituents and in vitro antioxidant activity of P. niruri leaf aqueous extract were investigated together with its effect on oxidative stress and antioxidant enzymes levels in diabetic rat kidney. Results. Treatment of diabetic male rats with P. niruri leaf aqueous extract (200 and 400 mg/kg) for 28 consecutive days prevents the increase in the amount of lipid peroxidation (LPO) product, malondialdehyde (MDA), and the diminution of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity levels in the kidney of diabetic rats. The amount of LPO showed strong negative correlation with SOD, CAT, and GPx activity levels. P. niruri leaf aqueous extract exhibits in vitro antioxidant activity with IC50 slightly lower than ascorbic acid. Phytochemical screening of plant extract indicates the presence of polyphenols. Conclusion. P. niruri leaf extract protects the kidney from oxidative stress induced by diabetes.


Positive and negative consequences with regard to cancer during adolescence. Experiences two years after diagnosis.
The purpose was to explore negative and positive consequences of cancer during adolescence experienced two years after diagnosis. Two years after diagnosis 38 persons, 15-21 years old, were asked two questions over the telephone: What, if anything, is bad for you due to the cancer disease? and What, if anything, is good for you due to the cancer disease? The answers were analysed by content analysis. Four categories of negative experiences were identified: a problematic body; unpleasant thoughts and feelings; outside the circle of friends; and difficulties with schoolwork. Six categories of positive experiences were identified: a more positive view of life; good self-esteem; knowledge and experience with regard to disease and hospital care; good relations; broader perspectives; and material gains. Two years after diagnosis those struck by cancer during adolescence experience not only a number of negative, but also positive, consequences of the cancer disease and its treatment.


An integrated physical and genetic map of the rice genome.
Rice was chosen as a model organism for genome sequencing because of its economic importance, small genome size, and syntenic relationship with other cereal species. We have constructed a bacterial artificial chromosome fingerprint-based physical map of the rice genome to facilitate the whole-genome sequencing of rice. Most of the rice genome ( approximately 90.6%) was anchored genetically by overgo hybridization, DNA gel blot hybridization, and in silico anchoring. Genome sequencing data also were integrated into the rice physical map. Comparison of the genetic and physical maps reveals that recombination is suppressed severely in centromeric regions as well as on the short arms of chromosomes 4 and 10. This integrated high-resolution physical map of the rice genome will greatly facilitate whole-genome sequencing by helping to identify a minimum tiling path of clones to sequence. Furthermore, the physical map will aid map-based cloning of agronomically important genes and will provide an important tool for the comparative analysis of grass genomes.


Masking of low frequency information in short-range apparent motion.
When an array of random dots is displaced, the ability to report the direction of apparent motion is subject to an upper spatial limit (dmax). Using spatially low-pass filtered random dot kinematograms we show that dmax is dependent on the upper cut-off frequency of the stimulus (Fh). The extent of this dependence is critically dependent on the size of the stimulus. Our results suggest a process whereby low spatial frequency motion information is masked by the presence of high spatial frequencies in the same region of the field, analogous to phenomena occurring in the perception of static form (e.g. the Abraham Lincoln effect). The effects of stimulus size on dmax, found for broad-band stimuli by ourselves and others, result from a loss of high frequency sensitivity at increased retinal eccentricities; this loss reduces the masking effect of high frequencies, as stimulus size increases.


Explaining direct care resource use of nursing home residents: findings from time studies in four states.
To explain variation in direct care resource use (RU) of nursing home residents based on the Resource Utilization Groups III (RUG-III) classification system and other resident- and unit-level explanatory variables. Primary data were collected on 5,314 nursing home residents in 156 nursing units in 105 facilities from four states (CO, IN, MN, MS) from 1998 to 2004. Study Design. Nurses and other direct care staff recorded resident-specific and other time caring for all residents on sampled nursing units. Care time was linked to resident data from the Minimum Data Set assessment instrument. Major variables were: RUG-III group (34-group), other health and functional conditions, licensed and other professional minutes per day, unlicensed minutes per day, and direct care RU (wage-weighted minutes). Resident- and unit-level relationships were examined through hierarchical linear modeling. Time study data were recorded with hand-held computers, verified for accuracy by project staff at the data collection sites and then merged into resident and unit-level data sets. Resident care time and RU varied between and within nursing units. RUG-III group was related to RU; variables such as length of stay and unit percentage of high acuity residents also were significantly related. Case-mix indices (CMIs) constructed from study data displayed much less variation across RUG-III groups than CMIs from earlier time studies. Results from earlier time studies may not be representative of care patterns of Medicaid and private pay residents. New RUG-III CMIs should be developed to better reflect the relative costs of caring for these residents.


Does IGFR1 inhibition result in increased muscle mass loss in patients undergoing treatment for pancreatic cancer?
IGF-1 plays a role in the growth of multiple tumor types, including pancreatic cancer. IGF-1 also serves as a growth factor for muscle. The impact of therapeutic targeting of IGF-1 on muscle mass is unknown. We evaluated muscle mass at L3 in patients enrolled in a randomized phase II study of MK-0646 (M), a monoclonal antibody directed against the IGF-1 protein, in patients with metastatic pancreatic cancer (MPC). Two different doses of M were tested, 5 and 10 mg/kg. We used the Slice-o-matic (ver 4.3) software to segregate CT images into muscle and fat components and measured muscle area (cm(2)) at baseline and after 2 and 4 months of treatment. Patients received either gemcitabine with erlotinib (G + E), G + E + M, or G + M. Differences between the groups were compared using t tests. Fifty-three patients had both baseline and 2-month imaging available for analysis. Of these, 42 received M with their chemo, and 11 had G + E only. After 2 months of treatment, both groups demonstrated decrease in muscle mass. G + E patients lost 5.6 % of muscle mass; M patients lost 9.1 and 8.6 % after treatment with 5 and 10 mg/kg, respectively (p = 0.53). Patients demonstrating a response lost less muscle (median 4.6 %) than those with stable disease (9.6 %) and progressive disease (8.9 %, p = 0.14). Muscle retention from baseline to 2-month imaging, defined as loss of <6 cm(2) of muscle, correlated with better survival than those patients demonstrating a muscle loss (HR 0.51, p = 0.03). MPC patients can be expected to lose muscle mass even while having clinical benefit (PR or SD) from chemotherapy. Muscle loss correlated with a risk of study drop-out and death. There was a non-significant trend toward greater muscle mass loss in patients on anti-IGF-1R therapy. However, it is unclear if this loss translates into functional differences between patients.


Report of the Council on Scientific Affairs: ultrasonic imaging of the heart: report of the Ultrasonography Task Force.
The use of ultrasonography in cardiology has progressed so dramatically that not only is anatomic information available but information can also be derived about cardiac hemodynamics. Applications range from intravascular ultrasonic imaging of coronary atherosclerosis to predictions of the severity of fetal valvular pulmonic stenosis detected in utero. We reviewed cardiac ultrasonography as utilized in B-mode imaging, pulsed and continuous-wave spectral Doppler, and Doppler color flow mapping. We reviewed specialized areas, including stress echo for wall motion analysis, valvular and congenital heart disease applications, and new applications in intraoperative, transesophageal, contrast echography, coronary imaging, and fetal echocardiography. Finally, future applications of quantitative flow mapping and intraluminal and interventional ultrasonography were considered along with the required technological advances.


Modulated Zika virus NS1 conjugate offers advantages for accurate detection of Zika virus specific antibody in double antigen binding and Ig capture enzyme immunoassays.
The accurate diagnosis and seroprevalence investigations of Zika virus (ZKV) infections remain complex due to cross reactivity with other flaviviruses. Two assay formats, both using labelled Zika virus NS1 antigen as a revealing agent (a double antigen binding assay, DABA, and an immunoglobulin Ig capture assay, G capture) were initially developed and compared with the indirect EuroimmunZ assay for the detection of anti-Zika antibody. Of 147 pre-Zika period serum samples, 39 (27%) were reactive in the EuroimmunZ or the DABA assays, 28 sera concordantly so. Such false reactivity was influenced by the serotype of Dengue virus (DV) to which individuals had been exposed to. Thus, of sera from patients undergoing secondary Dengue virus infection of known serotype, 91%, 45% and 28% of Dengue virus serotype 2, 3 and 4 respectively were reactive in one or more of the three assays. A novel method of quenching false sero-reactivity was therefore developed for the DABA and G capture assays. Initial addition of a single homologous Dengue virus serotype 3 NS1Ag quench significantly ablated false reactivities in the pre-Zika period sera. An equipotent quadrivalent quench comprising homologous Dengue virus serotypes 1 to 4 NS1Ag was shown to be optimum yet retained sensitivity for the detection of specific anti-Zika antibody. Comparing DABA and G capture assays using quenched and unquenched conjugates in comparison with EuroimmunZ early in the course of PCR-confirmed infection indicated that a significant component of the apparent early anti-ZIKA antibody response is likely to be due to a Zika virus-driven anamnestic anti-Dengue virus response. The increased specificity provided by homologous antigen quenching is likely to provide a significant improvement in sero-diagnostics and to be of clinical value.


Retargeting vesicular stomatitis virus using measles virus envelope glycoproteins.
Oncolytic vesicular stomatitis virus (VSV) has potent antitumor activity, but infects a broad range of cell types. Here, we used the measles virus (MV) hemagglutinin (H) and fusion (F) envelope glycoproteins to redirect VSV entry and infection specifically to tumor-associated receptors. Replication-defective VSV, deleted of its glycoprotein gene (VSVΔG), was pseudotyped with MV-F and MV-H displaying single-chain antibodies (scFv) specific for epidermal growth factor receptor (EGFR), folate receptor (FR), or prostate membrane-specific antigen (PSMA). Viral titers were ∼10(5) PFU/ml, but could be concentrated to 10(7) PFU/ml. Immunoblotting confirmed incorporation of the MV-H-scFv and MV-F into functional VSV virions. Although VSV-G was able to infect all tumor cell lines tested, the retargeted VSV infected only cells that expressed the targeted receptor. In vivo specificities of the EGFR-, FR-, and PSMA-retargeted VSV were assessed by intratumoral injection into human tumor xenografts. Analysis of green fluorescent protein reporter gene expression indicated that VSV infection was restricted to receptor-positive tumors. In summary, we have demonstrated for the first time that VSV can be efficiently retargeted to different cellular receptors using the measles display technology, yielding retargeted VSV vectors that are highly specific for tumors that express the relevant receptor.


A novel endoscopic treatment for ureteric remnant hemorrhage post laparoscopic radical nephrectomy.
Laparoscopic radical nephrectomy is commonly used to treat renal masses. Given the ubiquitous presence of this technique, rare complications are becoming more commonplace. It is thus essential that practicing urologists be aware of all possible complications as well as the novel management approaches that exist. This report presents a situation in which a patient developed rapid-onset, postoperative gross hematuria. This complication is rare and multiple sources of bleeding must be considered. In the situation reported here, the ureteric remnant was the cause of the unremitting gross hematuria. While others have described surgical exploration as the primary treatment, the authors were successful in using a minimally invasive endoscopic approach with fulguration and instillation of a fibrin sealant. Indeed, they propose that the endoscopic approach described herein may be considered first line in cases of unremitting gross hematuria originating from the ureteric remnant.


Global distributions and strain diversity of avian infectious bronchitis virus: a review.
The poultry industry faces challenge amidst global food security crisis. Infectious bronchitis is one of the most important viral infections that cause huge economic loss to the poultry industry worldwide. The causative agent, infectious bronchitis virus (IBV) is an RNA virus with great ability for mutation and recombination; thus, capable of generating new virus strains that are difficult to control. There are many IBV strains found worldwide, including the Massachusetts, 4/91, D274, and QX-like strains that can be grouped under the classic or variant serotypes. Currently, information on the epidemiology, strain diversity, and global distribution of IBV has not been comprehensively reported. This review is an update of current knowledge on the distribution, genetic relationship, and diversity of the IBV strains found worldwide.


Distinct and common expression of receptors for inflammatory mediators in vagal nodose versus jugular capsaicin-sensitive/TRPV1-positive neurons detected by low input RNA sequencing.
Capsaicin-sensitive sensory C-fibers derived from vagal ganglia innervate the visceral organs, and respond to inflammatory mediators and noxious stimuli. These neurons play an important role in maintenance of visceral homeostasis, and contribute to the symptoms of visceral inflammatory diseases. Vagal sensory neurons are located in two ganglia, the jugular ganglia (derived from the neural crest), and the nodose ganglia (from the epibranchial placodes). The functional difference, especially in response to immune mediators, between jugular and nodose neurons is not fully understood. In this study, we microscopically isolated murine nodose and jugular capsaicin-sensitive / Trpv1-expressing C-fiber neurons and performed transcriptome profiling using ultra-low input RNA sequencing. RNAseq detected genes with significantly differential expression in jugular and nodose neurons, which were mostly involved in neural functions. Transcriptional regulators, including Cited1, Hoxb5 and Prdm12 showed distinct expression patterns in the two C-fiber neuronal populations. Common and specific expression of immune receptor proteins was characterized in each neuronal type. The expression of immune receptors that have received little or no attention from vagal sensory biologists is highlighted including receptors for certain chemokines (CXCLs), interleukins (IL-4) and interferons (IFNα, IFNγ). Stimulation of immune receptors with their cognate ligands led to activation of the C-fibers in isolated functional assays.


Activation of system L heterodimeric amino acid exchangers by intracellular substrates.
System L-type transport of large neutral amino acids is mediated by ubiquitous LAT1-4F2hc and epithelial LAT2-4F2hc. These heterodimers are thought to function as obligatory exchangers, but only influx properties have been studied in some detail up until now. Here we measured their intracellular substrate selectivity, affinity and exchange stoichiometry using the Xenopus oocyte expression system. Quantification of amino acid influx and efflux by HPLC demonstrated an obligatory amino acid exchange with 1:1 stoichiometry. Strong, differential trans-stimulations of amino acid influx by injected amino acids showed that the intracellular substrate availability limits the transport rate and that the efflux selectivity range resembles that of influx. Compared with high extracellular apparent affinities, LAT1- and LAT2-4F2hc displayed much lower intracellular apparent affinities (apparent K(m) in the millimolar range). Thus, the two system L amino acid transporters that are implicated in cell growth (LAT1-4F2hc) and transcellular transport (LAT2-4F2hc) are obligatory exchangers with relatively symmetrical substrate selectivities but strongly asymmetrical substrate affinities such that the intracellular amino acid concentration controls their activity.


Up-regulation of LncRNA SNHG20 Predicts Poor Prognosis in Hepatocellular Carcinoma.
Recent studies indicated that long noncoding RNAs (lncRNAs) played important regulatory roles in carcinogenesis and cancer progression. However, the contribution of small nucleolar RNA host gene 20 (SNHG20) to cancer development remains largely unknown. The aim of the study is to investigate the expression of SNHG20 and its clinical significance in hepatocellular carcinoma (HCC). Our results showed that the expression of SNHG20 was remarkably up-regulated in HCC tissues compared with adjacent non-tumor liver tissues from 49 fresh HCC samples (cohort 1) detected by quantitative reverse-transcription polymerase chain reaction (qRT-PCR, P = 0.004). The results were confirmed in 144 formalin-fixed, paraffin-embedded HCC tissues (cohort 2) by in situ hybridization (ISH). Meanwhile, the expression of SNHG20 was associated with tumor size (P = 0.027 for cohort 1 and P = 0.046 for cohort 2) and clinical stage (P = 0.027 for cohort 1 and P = 0.028 for cohort 2). Importantly, patients with high expression of SNHG20 had a shorter overall survival (OS, P < 0.001) and disease-free survival (DFS, P < 0.001) than those with low expression of SNHG20. Univatiate and multivariate analysis showed that SNHG20 was a significant and independent prognostic predictor for OS of HCC patients (hazard ratio = 3.985, 95% CI = 1.981-8.017, P < 0.001). In addition, a total of 331 HCC patients' data from the Caner Genome Atlas project (TCGA) were used to validate our findings. Consistently, the results from TCGA HCC cohort demonstrated that SNHG20 were overexpressed in HCC tissues compared with non-tumor liver tissues (P < 0.001). Patients with higher expression levels of SNHG20 had poorer OS (P = 0.021) and DFS (P < 0.001). Functionally, knockdown of SNHG20 in SK-Hep-1 cells significantly inhibited cellular proliferation, migration, and invasion. In conclusion, SNHG20, up-regulated in patients with HCC, may serve as an independent prognostic predictor for HCC patients.


Enhancing Psychosis-Spectrum Nosology Through an International Data Sharing Initiative.
The latent structure of schizotypy and psychosis-spectrum symptoms remains poorly understood. Furthermore, molecular genetic substrates are poorly defined, largely due to the substantial resources required to collect rich phenotypic data across diverse populations. Sample sizes of phenotypic studies are often insufficient for advanced structural equation modeling approaches. In the last 50 years, efforts in both psychiatry and psychological science have moved toward (1) a dimensional model of psychopathology (eg, the current Hierarchical Taxonomy of Psychopathology [HiTOP] initiative), (2) an integration of methods and measures across traits and units of analysis (eg, the RDoC initiative), and (3) powerful, impactful study designs maximizing sample size to detect subtle genomic variation relating to complex traits (the Psychiatric Genomics Consortium [PGC]). These movements are important to the future study of the psychosis spectrum, and to resolving heterogeneity with respect to instrument and population. The International Consortium of Schizotypy Research is composed of over 40 laboratories in 12 countries, and to date, members have compiled a body of schizotypy- and psychosis-related phenotype data from more than 30000 individuals. It has become apparent that compiling data into a protected, relational database and crowdsourcing analytic and data science expertise will result in significant enhancement of current research on the structure and biological substrates of the psychosis spectrum. The authors present a data-sharing infrastructure similar to that of the PGC, and a resource-sharing infrastructure similar to that of HiTOP. This report details the rationale and benefits of the phenotypic data collective and presents an open invitation for participation.


The Ecology and Evolutionary Dynamics of Meiotic Drive.
Meiotic drivers are genetic variants that selfishly manipulate the production of gametes to increase their own rate of transmission, often to the detriment of the rest of the genome and the individual that carries them. This genomic conflict potentially occurs whenever a diploid organism produces a haploid stage, and can have profound evolutionary impacts on gametogenesis, fertility, individual behaviour, mating system, population survival, and reproductive isolation. Multiple research teams are developing artificial drive systems for pest control, utilising the transmission advantage of drive to alter or exterminate target species. Here, we review current knowledge of how natural drive systems function, how drivers spread through natural populations, and the factors that limit their invasion.


Functional interaction of GATA1 with erythroid Krüppel-like factor and Sp1 at defined erythroid promoters.
GATA and CACC elements commonly are codistributed within the regulatory domains of a variety of erythroid genes. Using Drosophila S2 cells, the actions of GATA1, Sp1, and erythroid Kruppel-like factor (EKLF) at these elements within model erythroid promoters have been assessed. For each promoter studied (erythroid pyruvate kinase, glycophorin B, and a murine betamaj globin-derived construct, GCT) Sp1 and EKLF each activated transcription despite differences in CACC element sequence, orientation, and positioning. However, GATA1 acted in apparent cooperativity with Sp1 at the pyruvate kinase promoter; with EKLF at the betamaj globin-derived GCT promoter; and with either Sp1 or EKLF at the glycophorin B promoter. Thus, GATA1 may functionally interact with each of these Krüppel-like factors depending on promoter context; and at the GCT promoter, transcriptional activation by GATA1 and EKLF was > or = 10-fold higher than levels attributable to additive effects. The possibility that interactions between these activators may be direct was supported by the specific binding of baculoviral-expressed EKLF to GATA1. This report underlines the likelihood that discrete roles exist for Sp1 and EKLF in erythroid gene activation, and supports a mechanism of direct cooperativity for EKLF and GATA1 as coregulators.


Effects of pulse shaping in laser spectroscopy and nuclear magnetic resonance.
Pulsed excitation fields are routinely used in most laser and nuclear magnetic resonance (NMR) experiments. In the NMR case, constant amplitude (rectangular) pulses have traditionally been used; in laser spectroscopy the exact pulse shape is often unknown or changes from shot to shot. This article is an overview of the effects of radio-frequency and laser pulse shapes and the instrumental requirements for pulse shaping. NMR applications to selective excitation, solvent suppression, elimination of phase roll, and reduced power dissipation are discussed, as are optical applications to soliton generation, velocity selective excitation, and quantitative population transfer.


Surgical management of malignant melanotic nerve sheath tumors: an institutional experience and systematic review of the literature.
Malignant melanotic nerve sheath tumors are rare tumors characterized by neoplastic melanin-producing Schwann cells. In this study, the authors report their institution's experience in treating spinal and peripheral malignant melanotic nerve sheath tumors and compare their results with the literature. Data were collected from 8 patients who underwent surgical treatment for malignant melanotic nerve sheath tumors between 1996 and 2023 at Mayo Clinic and 63 patients from the literature. Time-to-event analyses were performed for the combined group of 71 cases to evaluate the risk of recurrence, metastasis, and death based on tumor location and type of treatment received. Unpaired 2-sample t-tests and Fisher's exact tests were used to determine statistical significance between groups. Between 1996 and 2023, 8 patients with malignant melanotic nerve sheath tumors underwent surgery at the authors' institution, while 63 patients were identified in the literature. The authors' patients and those in the literature had the same mean age at diagnosis (43 years). At the authors' institution, 5 patients (63%) experienced metastasis, 6 patients (75%) experienced long-term recurrence, and 5 patients (62.5%) died. In the literature, most patients (60.3%) were males, with a peak incidence between the 4th and 5th decades of life. Nineteen patients (31.1%) were diagnosed with Carney complex. Nerve root tumors accounted for most presentations (n = 39, 61.9%). Moreover, 24 patients (38.1%) had intradural lesions, with 54.2% (n = 13) being intramedullary and 45.8% (n = 11) extramedullary. Most patients underwent gross-total resection (GTR) (n = 41, 66.1%), followed by subtotal resection (STR) (n = 12, 19.4%), STR with radiation therapy (9.7%), and GTR with radiation therapy (4.8%). Sixteen patients (27.6%) experienced metastasis, 23 (39.7%) experienced recurrence, and 13 (22%) died. Kaplan-Meier analyses showed no significant differences among treatment approaches in terms of recurrence-free, metastasis-free, and overall survival (p > 0.05). Similar results were obtained when looking at the differences with respect to intradural versus nerve root location of the tumor (p > 0.05). Malignant melanotic nerve sheath tumors are rare tumors with a high potential for malignancy. They carry a dismal prognosis, with a pooled local recurrence rate of 42%, distant metastasis rate of 27%, and mortality rate of 26%. The findings from this study suggest a trend favoring the use of GTR alone or STR with radiation therapy over STR alone. Mortality was similar regardless, which highlights the need for the development of effective treatment options to improve survival in patients with melanotic schwannomas.


Identification and characterization of an L-type lectin from obscure puffer Takifugu obscurus in response to bacterial infection.
L-type lectins (LTLs) contain a carbohydrate recognition domain homologous to leguminous lectins, and have functions in selective protein trafficking, sorting and targeting in the secretory pathway of animals. In this study, a novel LTL, designated as ToERGIC-53, was cloned and identified from obscure puffer Takifugu obscurus. The open reading frame of ToERGIC-53 contained 1554 nucleotides encoding 517 amino acid residues. The deduced ToERGIC-53 protein consisted of a signal peptide, a leguminous lectin domain (LTLD), a coiled-coil region, and a transmembrane region. Quantitative real-time PCR showed that ToERGIC-53 was expressed in all examined tissues, with the highest expression level in the liver. The expression of ToERGIC-53 was significantly upregulated after infection with Vibrio harveyi and Staphylococcus aureus. Recombinant ToERGIC-53-LTLD (rToERGIC-53-LTLD) protein could not only agglutinate and bind to one Gram-positive bacterium (S. aureus) and three Gram-negative bacteria (V. harveyi, V. parahaemolyticus and Aeromonas hydrophila), but also bind to glycoconjugates on the surface of bacteria such as lipopolysaccharide, peptidoglycan, mannose and galactose. In addition, rToERGIC-53-LTLD inhibited the growth of bacteria in vitro. All these results suggested that ToERGIC-53 might be a pattern recognition receptor involved in antibacterial immune response of T. obscurus.


The metabolically healthy but obese phenotype is associated with lower plasma levels of persistent organic pollutants as compared to the metabolically abnormal obese phenotype.
Although obesity is strongly linked to insulin resistance and type 2 diabetes, a subset of obese individuals termed metabolically healthy but obese (MHO) appears relatively protected from the development of cardiometabolic complications. The origins of this metabolically healthy phenotype remain unclear. Recently, persistent organic pollutants (POPs) have emerged as potential endocrine disruptors. The aim of this study was to test the hypothesis that the MHO phenotype presents lower circulating levels of POPs as compared to the metabolically abnormal obese (MAO) phenotype. We conducted a cross-sectional study of 76 nondiabetic obese (body mass index ≥30 kg/m(2)) postmenopausal women. Plasma concentrations of 21 POPs as well as cardiometabolic risk factors were analyzed. For similar age, body mass index, and fat mass index, MHO women (n = 40) showed higher insulin sensitivity levels and a more favorable cardiometabolic profile than MAO women (n = 36), as evidenced by a 2-fold increase in glucose disposal rates measured by the hyperinsulinemic-euglycemic clamp (P = .001). Among 18 detectable pollutants measured, MAO women had higher plasma concentrations of 12 POPs (fold increase, 1.4-2.9; P < .001-.036). Logistic regression analyses showed that the prevalence of the MAO phenotype was significantly associated with higher levels of total dioxin- and non-dioxin-like polychlorinated biphenyls (odds ratio, 4.7; 95% confidence interval, 1.8-12.5; P = .002), as well as trans-nonachlor (odds ratio, 6.1; 95% CI, 2.2-16.4; P < .001). Our study demonstrates that the metabolically healthy and abnormal phenotypes have distinct plasma POP profiles.


Genetic and Molecular Regulation of Extrasynaptic GABA-A Receptors in the Brain: Therapeutic Insights for Epilepsy.
GABA-A receptors play a pivotal role in many brain diseases. Epilepsy is caused by acquired conditions and genetic defects in GABA receptor channels regulating neuronal excitability in the brain. The latter is referred to as GABA channelopathies. In the last two decades, major advances have been made in the genetics of epilepsy. The presence of specific GABAergic genetic abnormalities leading to some of the classic epileptic syndromes has been identified. Advances in molecular cloning and recombinant systems have helped characterize mutations in GABA-A receptor subunit genes in clinical neurology. GABA-A receptors are the prime targets for neurosteroids (NSs). However, GABA-A receptors are not static but undergo rapid changes in their number or composition in response to the neuroendocrine milieu. This review describes the recent advances in the genetic and neuroendocrine control of extrasynaptic and synaptic GABA-A receptors in epilepsy and its impact on neurologic conditions. It highlights the current knowledge of GABA genetics in epilepsy, with an emphasis on the neuroendocrine regulation of extrasynaptic GABA-A receptors in network excitability and seizure susceptibility. Recent advances in molecular regulation of extrasynaptic GABA-A receptor-mediated tonic inhibition are providing unique new therapeutic approaches for epilepsy, status epilepticus, and certain brain disorders. The discovery of an extrasynaptic molecular mechanism represents a milestone for developing novel therapies such as NS replacement therapy for catamenial epilepsy.


Relationship of ventricular ectopy in men without apparent heart disease to occurrence of ischemic heart disease and sudden death.
The purpose of this investigation was to determine whether ventricular ectopic beats, or ventricular premature beats (VPBs), on routine electrocardiograms in men without apparent heart disease predict the later occurrence of clinical manifestations of ischemic heart disease (IHD). The Manitoba Study cohort consisted of 3983 men predominantly between 25 and 34 years of age and free of IHD at entry. During the 29-year observation period, 401 persons without clinical evidence of heart disease had VPBs on an electrocardiogram at a routine examination. They were followed 10.8 +/- 0.5 (SEM) years and 13.5% (54 men) later manifested IHD. Age-specific total IHD incidence was significantly (p less than 0.05) greater for men 40 to 59 years of age at VPB occurrence compared to men of the same age without VPBs. The clinical manifestation with the strongest association with VPBs was sudden death. VPB characteristics of frequency, configuration, coupling interval, and postextrasystolic T-wave change did not distinguish those who developed IHD. Prematurity index (R-R'/QT) showed a trend toward an association of late coupled ectopic beats (R-R'/QT greater than 1.6) and IHD risk. However, faster basic ventricular rate plus VPBs significantly correlated with greater IHD probability. Thus ventricular ectopic beats on a routine electrocardiogram in men over 40 years of age without apparent heart disease identify those at high risk for a clinical IHD event, especially sudden death.


The epidemiology of autism: a review.
There is some uncertainty about the rate and correlates of autism. Twenty-three epidemiological surveys of autism published in the English language between 1966 and 1998 were reviewed. Over 4 million subjects were surveyed; 1533 subjects with autism were identified. The methodological characteristics of each study are summarized, including case definition, case-finding procedures, participation rates and precision achieved. Across surveys, the median prevalence estimate was 5.2/10000. Half the surveys had 95% confidence intervals consistent with population estimates of 5.4-5.5/10000. Prevalence rates significantly increased with publication year, reflecting changes in case definition and improved recognition; the median rate was 7.2/10 000 for 11 surveys conducted since 1989. The average male/female ratio was 3.8:1, varying according to the absence or presence of mental retardation. Intellectual functioning within the normal range was reported in about 20% of subjects. On average, medical conditions of potential causal significance were found in 6% of subjects with autism, with tuberous sclerosis having a consistently strong association with autism. Social class and immigrant status did not appear to be associated with autism. There was no evidence for a secular increase in the incidence of autism. In eight surveys, rates for other forms of pervasive developmental disorders were two to three times higher than the rate for autism. Based on recent surveys, a minimum estimate of 18.7/10000 for all forms of pervasive developmental disorders was derived, which outlines the needs in special services for a large group of children.


Autoradiographic localization and characterization of tachykinin receptor binding sites in the rat brain and peripheral tissues.
Quantitative receptor autoradiography using several radiolabeled tachykinins was used to localize and characterize tachykinin peptide receptor binding sites in rat CNS and peripheral tissues. Autoradiographic localization and displacement experiments using several radiolabeled tachykinins indicate that in the rat there are at least 3 distinct tachykinin receptor binding sites. One of these is present in both the CNS and peripheral tissues, one is present only in the CNS, and one is present only in peripheral tissues. The first tachykinin receptor binding site, which is detectable in both the CNS and peripheral tissues, appears to prefer substance P (SP) as an endogenous ligand. Areas expressing high concentrations of this binding site include the medial septum, superior colliculus, inferior olive, inner plexiform layer of the retina, external muscle of the bladder, and the muscularis externa of the esophagus. The second type of tachykinin receptor binding site, which is detectable only in the CNS appears to prefer either neuromedin K (NK) and/or substance K (SK) as the endogenous ligand. This receptor binding site is labeled by Bolton-Hunter conjugates of NK, SK, eledoisin, or kassinin and is found in high concentrations in laminae 4 and 5 of the cerebral cortex, the ventral tegmental area, laminae 1 and 2 of the spinal cord, and the inner plexiform layer of the retina. The third type of tachykinin receptor binding site is detectable only in peripheral tissues and appears to prefer SK as the endogenous ligand. This receptor binding site is labeled by SK, eledoisin, or kassinin radioligands and tissues that express high concentrations include the muscularis mucosae of the esophagus, the circular muscle of the colon, and the external muscle of the bladder. These data suggest that SP receptors are expressed in the brain and peripheral tissues, NK receptors are expressed in the CNS, and SK receptors are expressed in peripheral tissue. These data fit well with radioimmunoassay data that suggest that, whereas in the CNS SP, SK and NK are present in high concentrations, in peripheral tissues only SP and SK are present in detectable concentrations. The present classification of tachykinin receptors places a lower limit on the number of mammalian tachykinin receptor types and provides a functional/morphological framework for exploring the diverse actions of tachykinin peptides in both the CNS and peripheral tissues.


Laboratory study of the effects of citric and ascorbic acids on injections prepared with brown heroin.
The addition of acidic substances to brown street heroin to facilitate the solubility of diamorphine in the injection preparation process is commonplace amongst UK injectors. Knowledge of the chemistry behind this process supports the need for this stage in the injection preparation process. It is currently illegal, under the Misuse of Drugs Act 1971, section 9A, to supply acidifiers and other paraphernalia to injectors in the UK. In the current climate of evidence-based practice, any consideration given to changing the law would look for evidence to illustrate that the paraphernalia was necessary. Although the theory behind the use of acidifiers suggests they are essential, no previous work using street heroin has actually been reported to illustrate this fact. Anecdotal information has found that drug users are being told by some service providers that the addition of acids is unnecessary. It is important that drugs services give credible information to their clients. The provision of inaccurate information in one area may lead to a lack of trust of all information provided. The small study reported here investigated, under controlled laboratory conditions, the effects of citric and ascorbic acids (vitamin C) on injections prepared with brown heroin, in order to demonstrate the need for acidifiers in the injection preparation process.


Computational and In Vitro Experimental Investigation of Intrathecal Drug Distribution: Parametric Study of the Effect of Injection Volume, Cerebrospinal Fluid Pulsatility, and Drug Uptake.
Intrathecal drug delivery is an attractive option to circumvent the blood-brain barrier for pain management through its increased efficacy of pain relief, reduction in adverse side effects, and cost-effectiveness. Unfortunately, there are limited guidelines for physicians to choose infusion or drug pump settings to administer therapeutic doses to specific regions of the spine or the brain. Although empiric trialing of intrathecal drugs is critical to determine the sustained side effects, currently there is no inexpensive in vitro method to guide the selection of spinal drug delivery parameters. The goal of this study is to demonstrate current computational capabilities to predict drug biodistribution while varying 3 parameters: (1) infusion settings, (2) drug chemistry, and (3) subject-specific anatomy and cerebrospinal fluid dynamics. We will discuss strategies to systematically optimize these 3 parameters to administer drug molecules to targeted tissue locations in the central nervous system. We acquired anatomical data from magnetic resonance imaging (MRI) and velocity measurements in the spinal cerebrospinal fluid with CINE-MRI for 2 subjects. A bench-top surrogate of the subject-specific central nervous system was constructed to match measured anatomical dimensions and volumes. We generated a computational mesh for the bench-top model. Idealized simulations of tracer distribution were compared with bench-top measurements for validation. Using reconstructions from MRI data, we also introduced a subject-specific computer model for predicting drug spread for the human volunteer. MRI velocity measurements at 3 spinal regions of interest reasonably matched the simulated flow fields in a subject-specific computer mesh. Comparison between the idealized spine computations and bench-top tracer distribution experiments demonstrate agreement of our drug transport predictions to this physical model. Simulated multibolus drug infusion theoretically localizes drug to the cervical and thoracic region. Continuous drug pump and single bolus injection were successful to target the lumbar spine in the simulations. The parenchyma might be targeted suitably by multiple boluses followed by a flush infusion. We present potential guidelines that take into account drug specific kinetics for tissue uptake, which influence the speed of drug dispersion in the model and potentially influence tissue targeting. We present potential guidelines considering drug-specific kinetics of tissue uptake, which determine the speed of drug dispersion and influence tissue targeting. However, there are limitations to this analysis in that the parameters were obtained from an idealized healthy patient in a supine position. The proposed methodology could assist physicians to select clinical infusion parameters for their patients and provide guidance to optimize treatment algorithms. In silico optimization of intrathecal drug delivery therapies presents the first steps toward a possible care paradigm in the future that is specific to personalized patient anatomy and diseases.


Application of microPET imaging approaches in the study of pediatric anesthetic-induced neuronal toxicity.
Advances in pediatric and obstetric surgery have resulted in an increase in the complexity, duration and number of anesthetic procedures. Currently, the general anesthetics that are used most often have either NMDA receptor blocking or GABA receptor activating properties. It has been reported that prolonged exposure of the developing brain to a clinically relevant concentration of anesthetics that have NMDA antagonist or GABA-mimetic properties, and/or their combinations, resulted in an extensive abnormal pattern of neuroapoptosis, and subsequent cognitive deficits in animals. Molecular imaging using positron emission tomography (PET) is a leading modality for obtaining non- or minimally invasive in vivo measurements of multiple biological processes in various organs. The development of microPET imaging applications has provided the ability to collect sensitive and quantitative three-dimensional molecular information from the living brains of a variety of animals. The main aim of this review was to describe molecular imaging approaches that have been used in the study of pediatric anesthetic-induced neuronal toxicity.


Isolation, synthesis and characterization of ω-TRTX-Cc1a, a novel tarantula venom peptide that selectively targets L-type Cav channels.
Spider venoms are replete with peptidic ion channel modulators, often with novel subtype selectivity, making them a rich source of pharmacological tools and drug leads. In a search for subtype-selective blockers of voltage-gated calcium (CaV) channels, we isolated and characterized a novel 39-residue peptide, ω-TRTX-Cc1a (Cc1a), from the venom of the tarantula Citharischius crawshayi (now Pelinobius muticus). Cc1a is 67% identical to the spider toxin ω-TRTX-Hg1a, an inhibitor of CaV2.3 channels. We assembled Cc1a using a combination of Boc solid-phase peptide synthesis and native chemical ligation. Oxidative folding yielded two stable, slowly interconverting isomers. Cc1a preferentially inhibited Ba(2+) currents (IBa) mediated by L-type (CaV1.2 and CaV1.3) CaV channels heterologously expressed in Xenopus oocytes, with half-maximal inhibitory concentration (IC50) values of 825nM and 2.24μM, respectively. In rat dorsal root ganglion neurons, Cc1a inhibited IBa mediated by high voltage-activated CaV channels but did not affect low voltage-activated T-type CaV channels. Cc1a exhibited weak activity at NaV1.5 and NaV1.7 voltage-gated sodium (NaV) channels stably expressed in mammalian HEK or CHO cells, respectively. Experiments with modified Cc1a peptides, truncated at the N-terminus (ΔG1-E5) or C-terminus (ΔW35-V39), demonstrated that the N- and C-termini are important for voltage-gated ion channel modulation. We conclude that Cc1a represents a novel pharmacological tool for probing the structure and function of L-type CaV channels.


Process design for efficient and controlled drug incorporation into polymeric micelle carrier systems.
For the efficient and well-controlled incorporation of the anti cancer drug adriamycin (ADR) into the inner core of a thermo-responsive polymeric micelle carrier system, we have analyzed and optimized the incorporation procedure in this paper. A dialysis method was used for preparing the micelle solution and ADR incorporation simultaneously. Quantities of ADR and triethylamine (TEA) were varied and the effects of their quantities were analyzed. Solvent composition at the starting time of dialysis was also varied. The initial dialysis condition, solvent with 40% water, brought about the largest amount and yield of ADR incorporation. With the initial 40% water content, it was considered that the block polymers formed a micelle-like association with a swollen hydrophobic core. This swollen core may be suitable for a large amount of ADR incorporation, since this core, swollen by an organic solvent-water mixture, is expected to show a liquid-state character to allow ADR molecules entry into the cores. By starting the dialysis procedure at this 40% water content, this swollen core suitable for the ADR incorporation is considered to be maintained for a much longer period than a case starting with a polymer-ADR solution in a solvent with a water content of less than 40%, and, therefore, ADR is expected to be incorporated efficiently. Preparation temperature of 20-25 degrees C was found to provide the most effective ADR incorporation in this thermo-responsive polymeric micelle system. These results indicate that the efficient incorporation of ADR can be achieved in consideration of the dynamic micelle formation and drug incorporation processes.


The development of whole blood titanium levels after instrumented spinal fusion - is there a correlation between the number of fused segments and titanium levels?
Most modern spinal implants contain titanium and remain in the patient's body permanently. Local and systemic effects such as tissue necrosis, osteolysis and malignant cell transformation caused by implants have been described. Increasing tissue concentration and whole blood levels of ions are necessary before a disease caused by a contaminant develops. The aim of the present study was the measurement of whole blood titanium levels and the evaluation of a possible correlation between these changes and the number of fused segments. A prospective study was designed to determine changes in whole blood titanium levels after spinal fusion and to analyze the correlation to the number of pedicle screws, cross connectors and interbody devices implanted.Blood samples were taken preoperatively in group I (n = 15), on the first, second and 10th day postoperatively, as well as 3 and 12 months after surgery.Group II (n = 16) served as a control group of volunteers who did not have any metal implants in the body. Blood samples were taken once in this group.The number of screw-rod-connections and the length of the spinal fusion were determined using radiographic pictures. This study was checked and approved by the ethical committee of the University of Tuebingen. The mean age in group I was 47 ± 22 years (range 16 - 85 years). There were three male (20%) and twelve female (80%) patients. The median number of fused segments was 5 (range 1 to 11 segments).No statistically significant increase in the titanium level was seen 12 months after surgery (mean difference: -7.2 μg/l, 95% CI: -26.9 to 12.5 μg/l, p = 0.446). By observing the individual titanium levels, 4 out of 15 patients demonstrated an increase in titanium levels 12 months after surgery.No statistically significant correlation between fused segments (r = -0.188, p = 0.503) length of instrumentation (r = -0.329, p = 0.231), number of interbody devices (r = -0.202, p = 0.291) and increase of titanium levels over the observation period was seen. Instrumented spinal fusion does not lead to a statistically significant increase in whole blood titanium levels. There seems to be no correlation between the number of pedicle screws, cross connectors and interbody devices implanted and the increase of serum titanium levels.


Malaria pathogenesis.
Malaria is a disease caused by repeated cycles of growth of the parasite Plasmodium in the erythrocyte. Various cellular and molecular strategies allow the parasite to evade the human immune response for many cycles of parasite multiplication. Under certain circumstances Plasmodium infection causes severe anemia or cerebral malaria; the expression of disease is influenced by both parasite and host factors, as exemplified by the exacerbation of disease during pregnancy. This article provides an overview of malaria pathogenesis, synthesizing the recent field, laboratory, and epidemiological data that will lead to the development of strategies to reduce mortality and morbidity.