Tactile Toe Agnosia and Percept of a "Missing Toe" in Healthy Humans.
A disturbance of body representation is central to many neurological and psychiatric conditions, but the mechanisms by which body representations are constructed by the brain are not fully understood. We demonstrate a directional disturbance in tactile identification of the toes in healthy humans. Nineteen young adult participants underwent tactile stimulation of the digits with the eyes closed and verbally reported the identity of the stimulated digit. In the majority of individuals, responses to the second and third toes were significantly biased toward the laterally neighboring digit. The directional bias was greater for the nondominant foot and was affected by the identity of the immediately preceding stimulated toe. Unexpectedly, 9/19 participants reported the subjective experience of a "missing toe" or "missing space" during the protocol. These findings challenge current models of somatosensory localization, as they cannot be explained simply by a lack of distinct representations for toes compared with fingers, or by overt toe-finger correspondences. We present a novel theory of equal spatial representations of digit width combined with a "preceding neighbor" effect to explain the observed phenomena. The diagnostic implications for neurological disorders that involve "digit agnosia" are discussed.


Characterization of compounds that induce symbiosis between sea anemone and anemone fish.
The species-specific partnership between the sea anemone and anemone fish in many parts of the Indo-Pacific region is a well-known phenomenon. Chemicals secreted by the sea anemone to elicit symbiotic behavior of the fish have been studied for two host-guest pairs, Radianthus kuekenthali (sea anemone)-Amphiprion perideraion (anemone fish) and Stoichactis kenti-A. ocellaris. A new pyridinium compound, amphikuemin, which induces characteristic attracted swimming (toward the chemical stimulus) in A. perideraion, has been isolated from R. kuekenthali and characterized. Several other chemicals that elicit characteristic symbiotic movements have also been identified.


Desmopressin and bleeding during invasive surgery.
Peri-operative bleeding is associated with invasive surgery and has traditionally been compensated for by blood transfusion. Concerns about the risk of transfusion-transmitted disease have led to an increasing interest in synthetic haemostatic agents. Desmopressin (1-deamino-8-D-arginine vasopressin), a synthetic analogue of vasopressin, has been shown to be of benefit in the peri-operative management of von Willebrand's disease or mild haemophilia A. This paper addresses the role of desmopressin and bleeding during invasive surgery, particularly during cardiopulmonary bypass. Clinical trials using desmopressin in open cardiac surgery indicate that it may reduce blood loss in those with an excessive bleeding tendency. However, it is difficult to identify this group pre-operatively.


Impact of a mobile application for heart rate assessment in simulated neonatal resuscitation: a randomised controlled cross-over study.
Clinical assessment of newborn heart rate (HR) at birth has been reported to be inaccurate. NeoTapAdvancedSupport (NeoTapAS) is a free-of-charge mobile application that showed good accuracy in HR estimation. This study aimed to evaluate the impact of NeoTapAS on timing of HR communication and resuscitation interventions. This was a randomised controlled cross-over (AB/BA) study evaluating HR assessment using auscultation plus NeoTapAS compared with auscultation plus mental computation in a high-fidelity simulated newborn resuscitation scenario. Twenty teams each including three paediatric residents were randomly assigned to AB or BA arms. The primary outcome was the timing of the first HR communication. Secondary outcomes included the timing of the following four HR communications and the timing of resuscitation interventions (positive pressure ventilation, chest compressions, intubation and administration of first dose of epinephrine). NeoTapAS reduced the time to the first HR communication (mean difference -13 s, 95% CI -23 to -4; p=0.009), and the time of initiation of chest compressions (mean difference -68 s, 95% CI -116 to -18; p=0.01) and administration of epinephrine (mean difference -76 s, 95% CI -115 to -37; p=0.0004) compared with mental computation. In a neonatal resuscitation simulated scenario, NeoTapAS reduced the time to the first HR communication and the time of initiation of chest compressions and administration of epinephrine compared with mental computation. This app can be especially useful in settings with limited availability of monitoring equipment, but further studies in clinical scenarios are warranted. NCT03730025.


Possible Northward Introgression of a Tropical Lineage of Rhipicephalus sanguineus Ticks at a Site of Emerging Rocky Mountain Spotted Fever.
Increasing rates of Rocky Mountain spotted fever (RMSF) in the southwestern United States and northern Mexico underscore the importance of studying the ecology of the brown dog tick, Rhipicephalus sanguineus, the vector in that region. This species is reported to comprise distinct tropical and temperate lineages that may differ in vectorial capacity for RMSF and are hypothesized to be limited in their geographical range by climatic conditions. In this study, lineage was determined for ticks from 9 locations in California, Arizona, and Mexico by DNA sequencing of 12S, 16S, and D-loop ribosomal RNA. As expected, sites in northern California and eastern Arizona had temperate-lineage ticks, and phylogenetic analysis revealed considerable genetic variability among these temperate-lineage ticks. However, tropical-lineage ticks extended north from Oaxaca, Mexico were well established along the entire border from San Diego, California to western Arizona, and were found as far north as Lytle Creek near Los Angeles, California (a site where both lineages were detected). Far less genetic variability in the tropical lineage despite the large geographical distances is supportive of a hypothesis of rapid northward expansion. Discovery of the tropical lineage north of the identified climatic limitations suggests that more work is needed to characterize this tick's ecology, vectorial capacity, expansion, possible evolution, and response to climate change.


Production of exoenzyme S during Pseudomonas aeruginosa infections of burned mice.
Antisera which distinguished between Pseudomonas aeruginosa exoenzyme S and toxin A neutralized the adenosine diphosphate ribosyl transferase activity of the homologous, but not the heterologous, enzyme. Skin extracts and sera from burned mice infected with the exoenzyme S-producing strain P. aeruginosa 388 contained adenosine diphosphate ribosyl transferase activity that was not found in skin extracts or sera from uninfected mice. On the basis of immunological reactivity and enzymatic properties, the adenosine diphosphate ribosyl transferase activity present in skin extracts and sera from P. aeruginosa 388-infected mice was identified as exoenzyme S. Active elongation factor 2 levels in tissues from strain 388-infected mice were normal at 24 h postinfection, indicating that strain 388 does not produce detectable amounts of toxin A in vivo. An unexpected finding in this investigation was the presence of exoenzyme S-inactivating activity in the sera from some nonimmunized animals.


Monitoring receptor-ligand interactions between surfaces by thermal fluctuations.
We describe a new method for determining receptor-ligand association/dissociation events across the interface of two surfaces (two-dimensional binding) by monitoring abrupt decrease/resumption in thermal fluctuations of a biomembrane force probe. Our method has been validated by rigorous control experiments and kinetic experiments. We show that cellular on-rate of association can be measured by analysis of intervals from a dissociation event to the next association event (waiting times). Similarly, off-rate of molecular dissociation can be measured by analysis of intervals from an association event to the next dissociation event (bond lifetimes). Different types of molecular bonds could be distinguished by different levels of reduction in thermal fluctuations. This novel method provides a powerful tool to study cell adhesion and signaling mediated by single or multiple receptor-ligand species.


Allyl sulfides are privileged substrates in aqueous cross-metathesis: application to site-selective protein modification.
Allyl sulfides undergo efficient cross-metathesis in aqueous media with Hoveyda-Grubbs second generation catalyst 1. The high reactivity of allyl sulfides in cross-metathesis was exploited in the first examples of cross-metathesis on a protein surface. S-Allylcysteine was incorporated chemically into the protein, providing the requisite allyl sulfide handle. Preliminary efforts to genetically incorporate S-allylcysteine into proteins are also reported.


Structure of the Saccharomyces cerevisiae HO gene and analysis of its upstream regulatory region.
The HO gene product of Saccharomyces cerevisiae is a site-specific endonuclease that initiates mating type interconversion. We have determined the nucleotide sequence of a 3,129-base-pair (bp) segment containing HO. The segment contains a single long open reading frame encoding a polypeptide of 586 amino acids, which has unusual (unbiased) codon usage and is preceded by 762 bp of upstream region. The predicted HO protein is basic (16% lysine and arginine) and is calculated to have a secondary structure that is 30% helical. The corresponding transcript is initiated approximately 50 nucleotides prior to the presumed initiation codon. Insertion of an Escherichia coli lacZ gene fragment into the putative HO coding segment inactivated HO and formed a hybrid HO-lacZ gene whose beta-galactosidase activity was regulated by the mating type locus in the same manner as HO (repressed by a 1-alpha 2). Upstream regions of 1,360 and 762 bp conferred strong repression; 436 bp led to partial constitutivity and 301 bp to full constitutivity. Thus, DNA sequences that confer repression of HO by a1-alpha 2 are at least 250 nucleotides upstream of the transcription start point and are within 436 nucleotides of the HO initiation codon. The progressive loss of repression suggests that both the -762 to -436 and the -436 to -301 intervals contain sites for regulation by a1-alpha 2. The HO gene contains two distinct regions that promote autonomous replication of plasmids in S. cerevisiae. These regions contain sequences that are homologous to the two conserved sequences that are associated with ARS activity.


Building workforce capacity for the surgical management of cervical cancer in a fragile, low-income African nation-Democratic Republic of the Congo.
Surgery is a cornerstone of the management of cervical cancer. Women diagnosed with cervical cancer in sub-Saharan Africa have very little access to specialised (gynaecologic oncology) surgical services. We describe our experiences and challenges of training local general gynaecologists to surgically treat early stage invasive cervical cancer at a private sector healthcare facility in a fragile, low-income African nation. Implementation of the training curriculum began with assigned self-directed learning. It continued with on-site training which consisted of preoperative surgical video reviews, pre- and intra-operative assessment of disease status, deconstruction of the designated surgical procedure into its critical subcomponents and trainees orally communicating the steps of the surgical procedure with the master trainers. High-volume repetition of a single surgical procedure over a short time interval, intra-operative bedside mentoring, post-operative case review and mental narration were critical to the process of surgical skills transfer. Nineteen radical abdominal hysterectomies were successfully performed over four training visits; trainees were able to perform the procedure alone after eight cases; surgical complications decreased over time. The trainees have continued to perform the surgical procedures independently. Life-saving surgical capacity for the treatment of cervical cancer has been established and sustained at a private sector healthcare facility in a fragile, low-income African setting, through an innovative model of surgical training.


Structural and Hydrogeological Controls on Hydrocarbon and Brine Migration into Drinking Water Aquifers in Southern New York.
Environmental concerns regarding the potential for drinking water contamination in shallow aquifers have accompanied unconventional energy development in the northern Appalachian Basin. These activities have also raised several critical questions about the hydrogeological parameters that control the naturally occurring presence and migration of hydrocarbon gases in shallow aquifers within petroliferous basins. To interrogate these factors, we analyzed the noble gas, dissolved ion, and hydrocarbon gas (molecular and isotopic composition) geochemistry of 98 groundwater samples from south-central New York. All samples were collected ≫1km from unconventional drilling activities and sample locations were intentionally targeted based on their proximity to various types of documented fault systems. In agreement with studies from other petroliferous basins, our results show significant correlations between elevated levels of radiogenic [4 He], thermogenic [CH4 ], and dissolved ions (e.g., Cl, Br, Sr, Ba). In combination, our data suggest that faults have facilitated the transport of exogenous hydrocarbon-rich brines from Devonian source rocks into overlying Upper Devonian aquifer lithologies over geologic time. These data conflict with previous reports, which conclude that hydrodynamic focusing regulates the occurrence of methane and salt in shallow aquifers and leads to elevated levels of these species in restricted flow zones within valley bottoms. Instead, our data suggest that faults in Paleozoic rocks play a fundamental role in gas and brine transport from depth, regulate the distribution of their occurrence in shallow aquifers, and influence the geochemistry of shallow groundwater in this petroliferous basin.


Use of formative research in developing a knowledge translation approach to rotavirus vaccine introduction in developing countries.
Rotavirus gastroenteritis is the leading cause of diarrheal disease mortality among children under five, resulting in 450,000 to 700,000 deaths each year, and another 2 million hospitalizations, mostly in the developing world. Nearly every child in the world is infected with rotavirus at least once before they are five years old. Vaccines to prevent rotavirus or minimize its severity are now becoming available, and have already been introduced into the public vaccine programs of several Latin American countries. The World Health Organization (WHO) has made rotavirus vaccine introduction in developing countries a high priority. The WHOs Guidelines for Vaccine Introduction indicates that a key determinant to achieving vaccine introduction is the public health priority of the disease, suggesting that where the disease is not a priority uptake of the vaccine is unlikely. WHO recommends conducting a qualitative analysis of opinions held by the public health community to determine the perceptions of the disease and the priority given to the vaccine. This paper presents the formative research results of a qualitative survey of public health providers in five low- and middle-income countries to determine if and to what degree rotavirus is perceived to be a problem and the priority of a vaccine. Open-ended surveys were carried out through focus group discussions and one-on-one interviews. Researchers discovered that in all five countries knowledge of rotavirus was extremely low, and as a result was not considered a high priority. However, diarrhea among young children was considered a high priority among public health providers in the three poorest countries with relatively high levels of child mortality: India, Indonesia, and Nicaragua. In the poorest countries, advocacy and communication efforts to raise awareness about rotavirus sufficient for prioritization and accelerated vaccine introduction might benefit from a knowledge translation approach that delivers information and evidence about rotavirus through the broader context of diarrheal disease control, an existing priority, and including information about other new interventions, specifically low-osmolarity oral rehydration solution and zinc treatment.


Internal sphincterotomy reduces postoperative pain after Milligan Morgan haemorrhoidectomy.
Over the last few years, there has been increasing attention on surgical procedures to treat haemorrhoids. The Milligan-Morgan haemorrhoidectomy is still one of the most popular surgical treatments of haemorrhoids. The aim of the present work is to assess postoperative pain, together with other early and late complications, after Milligan-Morgan haemorrhoidectomy as we could observe in our experience before and after performing an internal sphincterotomy. from January 1980 to May 2007, we operated 850 patients, but only 699 patients (median age 53) were included in the present study because they satisfied our inclusion criteria. The patients were divided into two groups: all the patients operated on before 1995 (group A); all the patients operated on after 1995 (group B). Since 1995 an internal sphincterotomy of about 1 cm has been performed at the end of the procedure. The data concerning the complications of these two groups were compared. All the patients received a check-up at one and six months after operation and a telephone questionnaire three years after operation to evalue medium and long term results. after one month 507 patients (72.5%) did not have any postoperative complication. Only 192 patients (27.46%) out of 699 presented postoperative complication and the most frequent one (23.03%) was pain. The number of patients who suffered from postoperative pain decreased significantly when performing internal sphincterotomy, going from 28.8% down to 10.45% (chi(2): 10,880; p = 0,0001); 95% Confidence Interval (CI) 24.7 to 28.9 (group A) and 10.17 to 10.72 (group B). In 51 cases (7.29%) urinary retention was registered. Six cases of bleeding (0.85%) were registered. Medium and long term follow up did not show any difference among the two groups. internal sphincterotomy: reduces significantly pain only in the first postoperative period, but not in the medium-long term follow up; does not increase the incidence of continence impairment when performed; does not influence the incidence of the other postoperative complications especially as regard medium and long term results.


Prevalence of attention deficit hyperactivity disorder and comorbid psychiatric and behavioral problems among primary school students in western Saudi Arabia.
To determine the prevalence of attention deficit hyperactivity disorder (ADHD), subtypes of ADHD, and psychiatric, academic, and behavioral comorbidity in public primary school students in Jeddah, Saudi Arabia. Methods: This is a cross-sectional study. A simple random sample of 6 primary government schools in Jeddah, Saudi Arabia, was identified (3 male, 3 female), and a random sample of classes in each of grades 1-6 were selected. Between July and November 2016, teachers in these classes were asked to complete the Vanderbilt ADHD scale on all students in their classes. Results: A total of 929 students were screened. The overall prevalence of ADHD was 5% (5.3% in girls, 4.7% in boys). The most prevalent subtype of ADHD was combined type (2.7%), followed by hyperactive type (1.2%), and inattentive type (1.1%). The highest prevalence of ADHD overall was in grade 3 (7.1%) and the lowest prevalence in grade 6 (3.4%). Among students with ADHD, prevalence of comorbid psychiatric, academic, and behavioral problems was widespread (56.5% oppositional defiant disorder/conduct disorder, 54.4% impaired academic performance, 44.4% classroom behavioral problems, 41.3% depression/anxiety). Comorbid problems were especially prevalent in combined ADHD subtype and in boys. Conclusions: Attention deficit hyperactivity disorder is common in primary school children in Jeddah, and is associated with widespread psychiatric, academic, and behavioral problems, especially in boys. These findings have implications for the diagnosis and treatment of this serious neurobehavioral disorder.


A soybean acyl carrier protein, GmACP, is important for root nodule symbiosis.
Legumes (members of family Fabaceae) establish a symbiotic relationship with nitrogen-fixing soil bacteria (rhizobia) to overcome nitrogen source limitation. Single root hair epidermal cells serve as the entry point for bacteria to infect the host root, leading to development of a new organ, the nodule, which the bacteria colonize. In the present study, the putative role of a soybean acyl carrier protein (ACP), GmACP (Glyma18g47950), was examined in nodulation. ACP represent an essential cofactor protein in fatty acid biosynthesis. Phylogenetic analysis of plant ACP protein sequences showed that GmACP was classified in a legume-specific clade. Quantitative reverse-transcription polymerase chain reaction analysis demonstrated that GmACP was expressed in all soybean tissues but showed higher transcript accumulation in nodule tissue. RNA interference-mediated gene silencing of GmACP resulted in a significant reduction in nodule numbers on soybean transgenic roots. Fluorescent protein-labeled GmACP was localized to plastids in planta, the site of de novo fatty acid biosynthesis in plants. Analysis of the fatty acid content of root tissue silenced for GmACP expression, as determined by gas chromatography-mass spectrometry, showed an approximately 22% reduction, specifically in palmitic and stearic acid. Taken together, our data provide evidence that GmACP plays an important role in nodulation.


Patterns of energy intake in patients with cirrhosis and healthy volunteers.
Fuel utilization and N economy are optimized in patients with cirrhosis by provision of several small meals throughout the day and a late-night snack of complex carbohydrate. Currently, however, only limited information is available on the patterns of energy intake in patients with chronic liver disease. The aims of the present study were to determine the number of days required to undertake such an investigation and to observe the daily distribution of energy intake in this patient population. Eight patients with cirrhosis and eight matched healthy volunteers kept weighed dietary intake records for fifteen separate days over a 6-month period. The records were analysed for energy intake per hour and the number and size of energy intake episodes per 24 h calculated. Intake was verified against resting energy expenditure. Fourteen separate observational days were required to investigate the pattern of energy intake in the cirrhotic patients while 20 d were required for healthy volunteers. Considerable inter- and intrasubject variations in the number and size of energy intake episodes were observed in both the patients and healthy volunteers. However, no significant differences were observed between the mean total number of daily energy intake episodes (6.3 (SD 1.6) v. 7.0 (SD 1.4)) or in the distribution of daily energy intake between the two groups. Most patients and volunteers tended to eat frequent small meals, often including a late-night snack, rather than two or three large meals daily. It should, therefore, be possible to establish optimum patterns of energy intake in these patients in line with recent guidelines.


Discovery of Plant Viruses From Tea Plant (Camellia sinensis (L.) O. Kuntze) by Metagenomic Sequencing.
The tea plant (Camellia sinensis (L.) O. Kuntze) is an economically important woody species. In this study, we collected 26 tea plant samples with typical discoloration symptoms from different tea gardens and performed metagenomic analysis based on next-generation sequencing. Homology annotation and PCR sequencing validation finally identified seven kinds of plant viruses from tea plant. Based on abundance distribution analysis, the two most abundant plant viruses were highlighted. Genetic characterization suggested that they are two novel virus species with relatively high homology to Blueberry necrotic ring blotch virus and American plum line pattern virus. We named the newly discovered viruses tea plant necrotic ring blotch virus (TPNRBV) and tea plant line pattern virus (TPLPV). Evolutionary relationship analysis indicated that TPNRBV and TPLPV should be grouped into the Blunervirus and the Ilarvirus genera, respectively. TPLPV might have same genome activation process with known ilarviruses based on sequence analysis. Moreover, specific primers for both viruses detection were designed and validated. The symptoms and ultrastructure of TPNRBV infected leaves were first recorded. Virus detections in the symptomatic and asymptomatic tissues from field plants showing tea plant necrotic ring blotch disease suggest that TPNRBV has a systemic movement feature. In summary, we first identified seven kinds of putative plant viruses by metagenomic analysis and report two novel viruses being latent pathogens to tea plant. The results will advance our understanding of tea plant virology and have significance for the genetic breeding of tea plants in the future.


Navigating complex labyrinths: optimal paths from chemical waves.
The properties of excitable media are exploited to find minimum-length paths in complex labyrinths. Optimal pathways are experimentally determined by the collection of time-lapse position information on chemical waves propagating through mazes prepared with the Belousov-Zhabotinsky reaction. The corresponding velocity fields provide maps of optimal paths from every point in an image grid to a particular target point. Collisions of waves that were temporarily separated by obstacles mark boundary lines between Significantly different paths with the same absolute distance. The pathfinding algorithm is tested in very complex mazes with a simple reaction-diffusion model.


Impact of breast structure on lesion detection in breast tomosynthesis, a simulation study.
This study aims to characterize the effect of background tissue density and heterogeneity on the detection of irregular masses in breast tomosynthesis, while demonstrating the capability of the sophisticated tools that can be used in the design, implementation, and performance analysis of virtual clinical trials (VCTs). Twenty breast phantoms from the extended cardiac-torso (XCAT) family, generated based on dedicated breast computed tomography of human subjects, were used to extract a total of 2173 volumes of interest (VOIs) from simulated tomosynthesis images. Five different lesions, modeled after human subject tomosynthesis images, were embedded in the breasts and combined with the lesion absent condition yielded a total of [Formula: see text] VOIs. Effects of background tissue density and heterogeneity on the detection of the lesions were studied by implementing a composite hypothesis signal detection paradigm with location known exactly, lesion known exactly or statistically, and background known statistically. Using the area under the receiver operating characteristic curve, detection performance deteriorated as density was increased, yielding findings consistent with clinical studies. A human observer study was performed on a subset of the simulated tomosynthesis images, confirming the detection performance trends with respect to density and serving as a validation of the implemented detector. Performance of the implemented detector varied substantially across the 20 breasts. Furthermore, background tissue density and heterogeneity affected the log-likelihood ratio test statistic differently under lesion absent and lesion present conditions. Therefore, considering background tissue variability in tissue models can change the outcomes of a VCT and is hence of crucial importance. The XCAT breast phantoms have the potential to address this concern by offering realistic modeling of background tissue variability based on a wide range of human subjects, comprising various breast shapes, sizes, and densities.


Actin and microtubule crosslinkers tune mobility and control co-localization in a composite cytoskeletal network.
Actin and microtubule filaments, with their auxiliary proteins, enable the cytoskeleton to carry out vital processes in the cell by tuning the organizational and mechanical properties of the network. Despite their critical importance and interactions in cells, we are only beginning to uncover information about the composite network. The challenge is due to the high complexity of combining actin, microtubules, and their hundreds of known associated proteins. Here, we use fluorescence microscopy, fluctuation, and cross-correlation analysis to examine the role of actin and microtubules in the presence of an antiparallel microtubule crosslinker, MAP65, and a generic, strong actin crosslinker, biotin-NeutrAvidin. For a fixed ratio of actin and microtubule filaments, we vary the amount of each crosslinker and measure the organization and fluctuations of the filaments. We find that the microtubule crosslinker plays the principle role in the organization of the system, while, actin crosslinking dictates the mobility of the filaments. We have previously demonstrated that the fluctuations of filaments are related to the mechanics, implying that actin crosslinking controls the mechanical properties of the network, independent of the microtubule-driven re-organization.


Determinants of utilisation differences for cancer medicines in Belgium, Scotland and Sweden.
Little comparative evidence is available on utilisation of cancer medicines in different countries and its determinants. The aim of this study was to develop a statistical model to test the correlation between utilisation and possible determinants in selected European countries. A sample of 31 medicines for cancer treatment that obtained EU-wide marketing authorisation between 2000 and 2012 was selected. Annual data on medicines' utilisation covering the in- and out-patient public sectors were obtained from national authorities between 2008 and 2013. Possible determinants of utilisation were extracted from HTA reports and complemented by contacts with key informants. A longitudinal mixed effect model was fitted to test possible determinants of medicines utilisation in Belgium, Scotland and Sweden. In the all-country model, the number of indications reimbursed positively correlated with increased consumption of medicines [one indication 2.6, 95% CI (1.8-3.6); two indications 2.4, 95% CI (1.4-4.3); three indications 4.9, 95% CI (2.2-10.9); all P < 0.01], years since EU-wide marketing authorisation [1.2, 95% CI (1.02-1.4); p < 0.05], price per DDD [0.9, 95% CI (0.998-0.999), P < 0.01], and Prescrire rating [0.5, 95% CI (0.3-0.9), P < 0.05] after adjusting for time and other covariates. In this study, the most important correlates of increased utilisation in a sample of cancer medicines introduced in the past 15 years were: medicines coverage and time since marketing authorisation. Prices had a negative effect on consumption in Belgium and Sweden. The positive impact of financial MEAs in Scotland suggests that the latter may remove the regressive effect of list prices on consumption.


Four-step enzyme-linked immunosorbent assay for detection of Treponema pallidum antibody.
Further studies of a four-step enzyme-linked immunosorbent assay procedure to detect Treponema pallidum antibody are described. High-titered antibody, produced in rabbits by intravenous injection of T. pallidum, was used to coat polyvinyl chloride microtiter plates. To these plates a known concentration of T. pallidum was added, followed in successive steps by serial dilutions of human sera and appropriately diluted peroxidase-labeled anti-human immunoglobulin G antibody. O-Phenylenediamine was the substrate. A total of 340 sera were obtained from the DeKalb County Sexually Transmitted Diseases Clinic, Atlanta, Ga., and examined within 3 days of receipt. Ninety-six percent test agreement between the enzyme-linked immunosorbent assay and the fluorescent treponemal antibody absorption-double staining test was obtained. A total of 372 additional sera stored at -20 degrees C were examined. The overall sensitivity of the enzyme-linked immunosorbent assay with sera from patients with various stages of syphilis was 96%. With sera from uninfected individuals, the specificity of the enzyme-linked immunosorbent assay was 95%. No antigen instability was noted with the two antigen preparations used during this evaluation.


Programmed death 1 is a marker of angioimmunoblastic T-cell lymphoma and B-cell small lymphocytic lymphoma/chronic lymphocytic leukemia.
Programmed death 1 (PD-1) is a lymphoid receptor that negatively regulates immune responses. PD-1 expression was recently reported in some T-cell non-Hodgkin lymphoma (NHL) subtypes, but the expression profile of PD-1 and its ligands (PD-L1 and PD-L2) in B-NHLs remains largely to be characterized. To investigate this issue, monoclonal antibodies against PD-1, PD-L1, and PD-L2 were generated by immunization of balb-c mice. A series of 161 lymphoma tissue and 11 blood samples was analyzed using either immunohistochemistry or flow cytometry. In reactive lymph nodes, PD-1 was mainly expressed in follicular T cells. In B-NHLs, PD-1 was mainly expressed in reactive T cells; but expression was also noted in neoplastic B cells from small lymphocytic lymphoma (SLL, 12/13), grade III follicular lymphoma (3/3), and diffuse large cell lymphoma (2/25). In contrast, neoplastic B cells from mantle cell lymphoma (0/11), marginal zone lymphoma (0/12), Burkitt lymphoma (0/3), and grade 1 to 2 follicular lymphoma (0/40) were PD-1 negative. PD-L1 and PD-L2 were negative in small B-cell lymphomas, including B-SLL. Flow cytometry showed that blood cells from chronic lymphocytic leukemia (B-CLL) also displayed PD-1 expression, which could be increased by CD40 stimulation. PD-1 expression in T-NHLs was restricted to the angioimmunoblastic subtype (8/8). These results show that PD-1 expression among B-NHLs is mainly associated with SLL/CLL and is influenced by activation of the CD40/CD40L pathway. Because the anti-PD-1.6.4 antibody works on paraffin sections, it represents a useful tool to differentiate SLL/CLL from other small B-cell lymphomas.


Complexes of Trypanosoma cruzi sterol 14α-demethylase (CYP51) with two pyridine-based drug candidates for Chagas disease: structural basis for pathogen selectivity.
Chagas disease, caused by the eukaryotic (protozoan) parasite Trypanosoma cruzi, is an alarming emerging global health problem with no clinical drugs available to treat the chronic stage. Azole inhibitors of sterol 14α-demethylase (CYP51) were proven effective against Chagas, and antifungal drugs posaconazole and ravuconazole have entered clinical trials in Spain, Bolivia, and Argentina. Here we present the x-ray structures of T. cruzi CYP51 in complexes with two alternative drug candidates, pyridine derivatives (S)-(4-chlorophenyl)-1-(4-(4-(trifluoromethyl)phenyl)-piperazin-1-yl)-2-(pyridin-3-yl)ethanone (UDO; Protein Data Bank code 3ZG2) and N-[4-(trifluoromethyl)phenyl]-N-[1-[5-(trifluoromethyl)-2-pyridyl]-4-piperi-dyl]pyridin-3-amine (UDD; Protein Data Bank code 3ZG3). These compounds have been developed by the Drugs for Neglected Diseases initiative (DNDi) and are highly promising antichagasic agents in both cellular and in vivo experiments. The binding parameters and inhibitory effects on sterol 14α-demethylase activity in reconstituted enzyme reactions confirmed UDO and UDD as potent and selective T. cruzi CYP51 inhibitors. Comparative analysis of the pyridine- and azole-bound CYP51 structures uncovered the features that make UDO and UDD T. cruzi CYP51-specific. The structures suggest that although a precise fit between the shape of the inhibitor molecules and T. cruzi CYP51 active site topology underlies their high inhibitory potency, a longer coordination bond between the catalytic heme iron and the pyridine nitrogen implies a weaker influence of pyridines on the iron reduction potential, which may be the basis for the observed selectivity of these compounds toward the target enzyme versus other cytochrome P450s, including human drug-metabolizing P450s. These findings may pave the way for the development of novel CYP51-targeted drugs with optimized metabolic properties that are very much needed for the treatment of human infections caused by eukaryotic microbial pathogens.


Trypanosoma vivax infections: pushing ahead with mouse models for the study of Nagana. II. Immunobiological dysfunctions.
Trypanosoma vivax is the main species involved in trypanosomosis, but very little is known about the immunobiology of the infective process caused by this parasite. Recently we undertook to further characterize the main parasitological, haematological and pathological characteristics of mouse models of T. vivax infection and noted severe anemia and thrombocytopenia coincident with rising parasitemia. To gain more insight into the organism's immunobiology, we studied lymphocyte populations in central (bone marrow) and peripherical (spleen and blood) tissues following mouse infection with T. vivax and showed that the immune system apparatus is affected both quantitatively and qualitatively. More precisely, after an initial increase that primarily involves CD4(+) T cells and macrophages, the number of splenic B cells decreases in a step-wise manner. Our results show that while infection triggers the activation and proliferation of Hematopoietic Stem Cells, Granulocyte-Monocyte, Common Myeloid and Megacaryocyte Erythrocyte progenitors decrease in number in the course of the infection. An in-depth analysis of B-cell progenitors also indicated that maturation of pro-B into pre-B precursors seems to be compromised. This interferes with the mature B cell dynamics and renewal in the periphery. Altogether, our results show that T. vivax induces profound immunological alterations in myeloid and lymphoid progenitors which may prevent adequate control of T. vivax trypanosomosis.


Curcumin inhibits cyclooxygenase-2 transcription in bile acid- and phorbol ester-treated human gastrointestinal epithelial cells.
We investigated whether curcumin, a chemopreventive agent, inhibited chenodeoxycholate (CD)- or phorbol ester (PMA)-mediated induction of cyclooxygenase-2 (COX-2) in several gastrointestinal cell lines (SK-GT-4, SCC450, IEC-18 and HCA-7). Treatment with curcumin suppressed CD- and PMA-mediated induction of COX-2 protein and synthesis of prostaglandin E2. Curcumin also suppressed the induction of COX-2 mRNA by CD and PMA. Nuclear run-offs revealed increased rates of COX-2 transcription after treatment with CD or PMA and these effects were inhibited by curcumin. Treatment with CD or PMA increased binding of AP-1 to DNA. This effect was also blocked by curcumin. In addition to the above effects on gene expression, we found that curcumin directly inhibited the activity of COX-2. These data provide new insights into the anticancer properties of curcumin.


Prevalence of the uncinate process, agger nasi cell and their relationship in a Taiwanese population.
The uncinate process (UP) and the agger nasi (AN) cell are both important anatomical landmarks in endoscopic sinus surgery. The superior attachment of the UP and the prevalence of the AN cell show great anatomical variability and affect the anatomy of the frontal recess. The aim of this study was to investigate the superior attachment types of the UP, the presence of the AN cell, and the relationship between the two. Two hundred sixty-four sides of 132 patients with identifiable superior attachments of the UP were selected from our 2007-08 computed tomography (CT) scan records. Exclusion criteria were previous nasal or paranasal sinus surgery, neoplastic disease, and a history of nasal trauma. The superior attachment type of the UP and the prevalence of the AN cell were evaluated. Three case series from the English-language literature were selected for racial comparison. Single insertion of the UP into the lamina papyracea was the most common type (70.4%). The other types of UP superior attachment were found to have the following distribution: 10.2% into the middle turbinate; 7.6% into the lamina papyracea and the skull base; 6.1% into the skull base, 4.9% into the lamina papyracea and the middle turbinate; and 0.8% into the middle turbinate and the skull base. The distribution of the UP insertion types with respect to ethnicity was statistically significant (P < 0.01). The AN cell was found in 216 (81.8%) of 264 sides. The AN cell was present bilaterally in 95.3% of the cases (206 sides, 103 patients) and unilaterally in 4.7% (10 sides). The prevalence of the AN cell according to superior attachment of the UP types was not statistically significant (chi(2) = 8.6, p = 0.126). The nasofrontal anatomy did show some variations among different races. The single superior attachment of the UP into the lamina papyracea had the highest prevalence. The AN cell was found to be present in 81.8% of cases. The relationship between the presence of the AN cell and the superior attachment types of the UP was not statistically significant.


Combination of agrin and laminin increase acetylcholine receptor clustering and enhance functional neuromuscular junction formation In vitro.
Clustering of acetylcholine receptors (AChR) at the postsynaptic membrane is a crucial step in the development of neuromuscular junctions (NMJ). During development and after denervation, aneural AChR clusters form on the sarcolemma. Recent studies suggest that these receptors are critical for guiding and initiating synaptogenesis. The aim of this study is to investigate the effect of agrin and laminin-1; agents with known AChR clustering activity; on NMJ formation and muscle maturation. Primary myoblasts were differentiated in vitro on collagen, laminin or collagen and laminin-coated surfaces in the presence or absence of agrin and laminin. The pretreated cells were then subject to innervation by PC12 cells. The number of neuromuscular junctions was assessed by immunocytochemical co-localization of AChR clusters and the presynaptic marker synaptophysin. Functional neuromuscular junctions were quantitated by analysis of the level of spontaneous as well as neuromuscular blocker responsive contractile activity and muscle maturation was assessed by the degree of myotube striation. Agrin alone did not prime muscle for innervation while a combination of agrin and laminin pretreatment increased the number of neuromuscular junctions formed and enhanced acetylcholine based neurotransmission and myotube striation. This study has direct clinical relevance for treatment of denervation injuries and creating functional neuromuscular constructs for muscle tissue repair.


Complementary striped expression patterns of NK homeobox genes during segment formation in the annelid Platynereis.
NK genes are related pan-metazoan homeobox genes. In the fruitfly, NK genes are clustered and involved in patterning various mesodermal derivatives during embryogenesis. It was therefore suggested that the NK cluster emerged in evolution as an ancestral mesodermal patterning cluster. To test this hypothesis, we cloned and analysed the expression patterns of the homologues of NK cluster genes Msx, NK4, NK3, Lbx, Tlx, NK1 and NK5 in the marine annelid Platynereis dumerilii, a representative of trochozoans, the third great branch of bilaterian animals alongside deuterostomes and ecdysozoans. We found that most of these genes are involved, as they are in the fly, in the specification of distinct mesodermal derivatives, notably subsets of muscle precursors. The expression of the homologue of NK4/tinman in the pulsatile dorsal vessel of Platynereis strongly supports the hypothesis that the vertebrate heart derived from a dorsal vessel relocated to a ventral position by D/V axis inversion in a chordate ancestor. Additionally and more surprisingly, NK4, Lbx, Msx, Tlx and NK1 orthologues are expressed in complementary sets of stripes in the ectoderm and/or mesoderm of forming segments, suggesting an involvement in the segment formation process. A potentially ancient role of the NK cluster genes in segment formation, unsuspected from vertebrate and fruitfly studies so far, now deserves to be investigated in other bilaterian species, especially non-insect arthropods and onychophorans.


History and Future of Nucleic Acid Amplification Technology Blood Donor Testing.
The introduction of blood donor screening by virus nucleic acid amplification technology (NAT) in the mid to late 1990s was driven by the so-called AIDS and hepatitis C virus (HCV) epidemic, with thousands of recipients of infected blood products and components. Plasma fractionators were the first to introduce NAT testing besides pathogen reduction procedures, to reduce the virus transmission risk through their products. To achieve a similar safety standard, NAT was then also introduced for labile blood components. German transfusion centres were the first to start in-house NAT testing of their donations in pools of up to 96 samples for HCV, hepatitis B virus (HBV), and human immunodeficiency virus-1 (HIV-1). Years later the diagnostics industry provided commercial HCV and HIV-1 and later HBV NAT tests on automated platforms. NAT tests for HIV-2, hepatitis A virus, and Parvovirus B19 followed, again driven by transfusion centres with their in-house tests. When severe acute respiratory syndrome corona virus (SARS-CoV) and West Nile Virus emerged it was the NAT that enabled the manufacturers and transfusion centres to instantly introduce sensitive and specific screening tests. Subsequent automation including sample preparation has significantly reduced the costs and complexity of the procedure and made it affordable to middle income countries as well. Currently more than 60 million donations per year are NAT tested worldwide and the remaining residual risk of virus transmission by blood components and products could be reduced to almost zero. Automation rendered possible the reduction of pool size in conjunction with increased throughput and sensitivity. Thus, antibody and antigen testing may be dispensable in the long run, particularly in the combination of NAT testing with pathogen reduction. There are new technologies on the horizon like digital droplet PCR, next-generation sequencing, lab-on-a-chip, and digital antigen assays, which are comparably sensitive. However, each of these has limitations, either in throughput, costs, automation, time to result, specificity, or the need for NAT as an integral part of the technology. Thus, NAT is still the shortest and most efficient means to the result. Donor screening NAT also contributed significantly to our knowledge on how fast viruses replicate, and on the respective diagnostic window. In conjunction with animal and patient studies, we have learned more about the minimal infectious dose and the epidemics in the donor population.


Characterization of human upper airway epithelial progenitors.
New epithelial cells are generated through the proliferation and differentiation of resident progenitor cells in the nasal cavity. In several upper airway diseases, such as cystic fibrosis and chronic rhinosinusitis, self-renewing progenitor cells may be functionally defective, or compromised in their ability, to regenerate cells that maintain normal mucociliary clearance. Herein, we describe our early work to define and characterize a rare population of human nasal epithelial putative progenitors. Single-cell suspensions of human ethmoid sinus tissues were prepared following endoscopic sinus surgery. Cell surface antibodies were analyzed as candidate markers for detecting progenitor cells. A panel of antibodies, including epithelial cell adhesion molecule (EpCAM, epithelial cells), CD45 (hematopoietic cells), nerve growth factor receptor (NGFR/CD271), intercellular adhesion molecule-1 (ICAM1/CD54), and integrin-α6 (ITGA6/CD49f) were used to resolve epithelial progenitor candidates by high-dimensional flow cytometry and the gating technique of fluorescence minus one (FMO) controls. A rare population of approximately 0.06% of total ethmoid cells was discriminated as EpCAM(-) CD45(-) NGFR(+) ICAM1(+) by surface markers. Use of ITGA6 was excluded based on FMO control analysis. This lineage-negative population was purified to 99% homogeneity by cell sorting and analyzed by immunofluorescence microscopy. Sorted cells were subsequently confirmed to uniformly express the transcription factor p63. Upon in vitro culture, lineage-negative clonal cells were confirmed to spontaneously differentiate into epithelial lineage-positive cells. Using the NGFR and ICAM1 cellular coordinates, we have identified a promising population of native human nasal epithelial progenitor cells that require more formal investigation for their role in upper airway regeneration.


From the exposome to mechanistic understanding of chemical-induced adverse effects.
The exposome encompasses an individual's exposure to exogenous chemicals, as well as endogenous chemicals that are produced or altered in response to external stressors. While the exposome concept has been established for human health, its principles can be extended to include broader ecological issues. The assessment of exposure is tightly interlinked with hazard assessment. Here, we explore if mechanistic understanding of the causal links between exposure and adverse effects on human health and the environment can be improved by integrating the exposome approach with the adverse outcome pathway (AOP) concept that structures and organizes the sequence of biological events from an initial molecular interaction of a chemical with a biological target to an adverse outcome. Complementing exposome research with the AOP concept may facilitate a mechanistic understanding of stress-induced adverse effects, examine the relative contributions from various components of the exposome, determine the primary risk drivers in complex mixtures, and promote an integrative assessment of chemical risks for both human and environmental health.


Membrane ionic current photomodification by rose bengal and menadione: role of singlet oxygen.
Photosensitized modification of ionic leak current and potassium current was studied in frog cardiac atrial cells using whole cell patch clamp techniques. Rose bengal (RB) and menadione (MQ) were used as photosensitizers. Separate photophysical studies of the photosensitizers in deuterium oxide solution demonstrated that MQ did not produce singlet oxygen as evidenced by the lack of luminescence at 1270 nm, whereas RB was an efficient singlet oxygen generator. Both photosensitizers sensitized block of potassium current in atrial cells, and both sensitized an increase of ionic leak current. However, when photosensitizer concentrations and illumination intensities were adjusted to match the rate of block of potassium current by the two photosensitizers, there were dramatic differences in leak current increase, both quantitatively and qualitatively. Menadione sensitized a much slower increase in leak current than did RB. Further, the leak current sensitized by MQ had a more positive reversal potential than that sensitized by RB, suggesting a less potassium-selective leak current pathway. The results suggest that, while the effects of singlet oxygen and non-singlet oxygen modification of cell membranes may be similar, there may also be significant differences in the resulting membrane permeabilities. The results also demonstrate that MQ and RB may be useful agents to study the role of singlet oxygen versus non-singlet oxygen modification of biological systems.


Upper Limb Neurodynamic Test 1 on Healthy Individuals: Intra- and Intersession Reliability of the Angle between Pain Onset and Submaximal Pain.
Assessment of nerve trunk mechanosensitivity using the upper limb neurodynamic test 1 (ULNT1) often includes measurement of the angle of occurrence in the range of pain onset (PO) and submaximal pain (SP). A measurement that better fits the idea of mechanosensitivity could be the angle between PO and SP (AbOS). This study investigated the intra- and intersession reliability of AbOS, PO, and SP during the ULNT1. Forty-four healthy volunteers underwent three ULNT1 to the point of PO and SP, twice in the first session and once in the second. AbOS, PO, and SP angles of occurrence reliability were examined using the Intraclass Correlation Coefficient (ICC 3,1) and Bland-Altman plots. The intra- and intersession ICC values for AbOS were 0.71 (95% CI: 0.47; 0.85) and 0.79 (95% CI: 0.60; 0.89), respectively. The intra- and intersession mean difference and 95% limits of agreement (±1.96 SD) in the Bland-Altman plots were 2.3° (-18.3°; 23.1°) and 2.8° (-14.7°; 20.4°), respectively. The intra- and intersession reliability of the AbOS during the ULNT1 in healthy individuals is high and higher than the reliability of PO and SP angles of occurrence. The AbOS could be a preferable variable in the assessment of neural mechanosensitivity.


Population Structure and Genetic Diversity Within the Endangered Species Pityopsis ruthii (Asteraceae).
Pityopsis ruthii (Ruth's golden aster) is a federally endangered herbaceous perennial endemic to the Hiwassee and Ocoee Rivers in southeastern Tennessee, United States. Comprehensive genetic studies providing novel information to conservationists for preservation of the species are lacking. Genetic variation and gene flow were evaluated for 814 individuals from 33 discrete locations using polymorphic microsatellites: seven chloroplast and twelve nuclear. A total of 198 alleles were detected with the nuclear loci and 79 alleles with the chloroplast loci. Gene flow was estimated, with the Hiwassee River (Nm = 2.16; FST = 0.15) showing higher levels of gene flow and lower levels of population differentiation than the Ocoee River (Nm = 1.28; FST = 0.19). Population structure was examined using Bayesian cluster analyses. Nuclear and chloroplast analyses were incongruent. From the chloroplast microsatellites, three clusters were identified; all were present in sampling sites at both rivers, indicating a lack of allele fixation along rivers. Nuclear markers revealed two clusters and separated by river. When the Hiwassee River locations were analyzed, four clusters were identified for both the chloroplast and nuclear microsatellites, though the individuals clustered differently. Analysis of the Ocoee River revealed two clusters for the chloroplast microsatellites and three for the nuclear microsatellites. We recommend P. ruthii be managed as four populations for the Hiwassee River and three populations for the Ocoee River. Our results provide critical genetic information for P. ruthii that can be used for species management decisions to drive future population augmentation/reintroduction and ex situ conservation efforts.


Promotion and Prevention Focused Feeding Strategies: Exploring the Effects on Healthy and Unhealthy Child Eating.
There is a general lack of research addressing the motivations behind parental use of various feeding practices. Therefore, the present work aims to extend the current literature on parent-child feeding interactions by integrating the traditional developmental psychological perspective on feeding practices with elements of Regulatory Focus Theory (RFT) derived from the field of motivational psychology. In this paper, we seek to explain associations between parental feeding practices and child (un)healthy eating behaviors by categorizing parental feeding practices into promotion and prevention focused strategies, thus exploring parent-child feeding interactions within the framework of RFT. Our analyses partly supported the idea that (1) child healthy eating is positively associated with feeding practices characterized as promotion focused, and (2) child unhealthy eating is negatively associated with feeding practices characterized as prevention focused. However, a general observation following from our results suggests that parents' major driving forces behind reducing children's consumption of unhealthy food items and increasing their consumption of healthy food items are strategies that motivate rather than restrict. In particular, parents' provision of a healthy home food environment seems to be essential for child eating.


Resistance to Plant-Parasitic Nematodes in Chickpea: Current Status and Future Perspectives.
Plant-parasitic nematodes constrain chickpea (Cicer arietinum) production, with annual yield losses estimated to be 14% of total global production. Nematode species causing significant economic damage in chickpea include root-knot nematodes (Meloidogyne artiella, M. incognita, and M. javanica), cyst nematode (Heterodera ciceri), and root-lesion nematode (Pratylenchus thornei). Reduced functionality of roots from nematode infestation leads to water stress and nutrient deficiency, which in turn lead to poor plant growth and reduced yield. Integration of resistant crops with appropriate agronomic practices is recognized as the safest and most practical, economic and effective control strategy for plant-parasitic nematodes. However, breeding for resistance to plant-parasitic nematodes has numerous challenges that originate from the narrow genetic diversity of the C. arietinum cultigen. While levels of resistance to M. artiella, H. ciceri, and P. thornei have been identified in wild Cicer species that are superior to resistance levels in the C. arietinum cultigen, barriers to interspecific hybridization restrict the use of these crop wild relatives, as sources of nematode resistance. Wild Cicer species of the primary genepool, C. reticulatum and C. echinospermum, are the only species that have been used to introgress resistance genes into the C. arietinum cultigen. The availability of genomic resources, including genome sequence and re-sequence information, the chickpea reference set and mini-core collections, and new wild Cicer collections, provide unprecedented opportunities for chickpea improvement. This review surveys progress in the identification of novel genetic sources of nematode resistance in international germplasm collections and recommends genome-assisted breeding strategies to accelerate introgression of nematode resistance into elite chickpea cultivars.


One-step continuous extrusion process for the manufacturing of solid dispersions.
The purpose of this study was to evaluate the performance of synthetic magnesium aluminometasilicate (MAS) as a novel inorganic carrier in hot melt extrusion (HME) processing of indomethacin (IND) for the development of solid dispersions. A continuous extrusion process at various IND/excipient blend ratios (20%, 30% and 40%) was performed using a twin-screw extruder. Physicochemical characterization carried out by SEM, DSC, and XRPD demonstrated the presence of IND in amorphous nature within the porous network of the inorganic material for all extruded formulations. Further, AFM and FTIR studies revealed a single-phase amorphous system and intermolecular H-bonding formation. The IND/MAS extrudates showed enhanced INM dissolution rates within 100% been released within 1h. Stability studies under accelerated conditions (40°C, RH 75%) showed that MAS retained the physical stability of the amorphous solid dispersions even at high drug loadings for 12 months.


Crystalline lens thickness determines the perceived chromatic difference in magnification.
Since the origin of the high interindividual variability of the chromatic difference in retinal image magnification (CDM) in the human eye is not well understood, optical parameters that might determine its magnitude were studied in 21 healthy subjects with ages ranging from 21 to 58 years. Two psychophysical procedures were used to quantify CDM. They produced highly correlated results. First, a red and a blue square, presented on a black screen, had to be matched in size by the subjects with their right eyes. Second, a filled red and blue square, flickering on top of each other at 2 Hz, had to be adjusted in perceived brightness and then in size to minimize the impression of flicker. CDM varied widely among subjects from 0.0% to 3.6%. Biometric ocular parameters were measured with low coherence interferometry and crystalline lens tilt and decentration with a custom-built Purkinjemeter. Correlations were studied between CDM and corneal power, anterior chamber depth, lens thickness, lens tilt and lens decentration, and vitreous chamber depths. Lens thickness was found significantly correlated with CDM and accounted for 64% of its variance. Vertical lens tilt and decentration were also significantly correlated. It was also found that CDM increased by 3.5% per year, and part of this change can be attributed to the age-related increase in lens thickness.


Readily functionalized AAA-DDD triply hydrogen-bonded motifs.
Herein we present a new, readily functionalized AAA-DDD hydrogen bond array. A novel AAA monomeric unit (3a-b) was obtained from a two-step synthetic procedure starting with 2-aminonicotinaldehyde via microwave radiation (overall yield of 52-66%). 1H NMR and fluorescence spectroscopy confirmed the complexation event with a calculated association constant of 1.57 × 107 M-1. Likewise, the usefulness of this triple hydrogen bond motif in supramolecular polymerization was demonstrated through viscosity measurements in a crosslinked supramolecular alternating copolymer.


Pharmacologic modulation of periprosthetic osteolysis.
Wear and periprosthetic osteolysis of total joint replacements continue to be the most important problems in arthroplasty surgery. Despite the introduction of improved technologies including alternative bearing surfaces for TJRs, wear is inevitable because of relative movement at different interfaces and processes such as electrolysis and material degradation. Worn, clinically failing implants need to be followed closely and revised when appropriate. However, early wear and minor osteolysis do not result necessarily in progressive failure of the prosthesis. Indeed such cases may be followed up clinically and radiographically to establish the functional and biologic sequelae of wear and the timeline of these events. This scenario provides an opportunity to modulate the adverse biologic reaction associated with wear particles that includes chronic inflammation, the foreign body response, and periprosthetic bone destruction. Currently, immunological events associated with wear particles are becoming understood more clearly. Strategies to mitigate adverse processes associated with wear debris include local or systemic administration of immune modulators, signaling molecules, anti-inflammatory agents and growth factors, and altering osteoclast function. Ultimately, prevention of accelerated wear and periprosthetic osteolysis will be achieved with improved bearing surfaces and prosthetic designs.


Synthesizing Quantitative Evidence for Evidence-based Nursing: Systematic Review.
As evidence-based practice has become an important issue in healthcare settings, the educational needs for knowledge and skills for the generation and utilization of healthcare evidence are increasing. Systematic review (SR), a way of evidence generation, is a synthesis of primary scientific evidence, which summarizes the best evidence on a specific clinical question using a transparent, a priori protocol driven approach. SR methodology requires a critical appraisal of primary studies, data extraction in a reliable and repeatable way, and examination for validity of the results. SRs are considered hierarchically as the highest form of evidence as they are a systematic search, identification, and summarization of the available evidence to answer a focused clinical question with particular attention to the methodological quality of studies or the credibility of opinion and text. The purpose of this paper is to introduce an overview of the fundamental knowledge, principals and processes in SR. The focus of this paper is on SR especially for the synthesis of quantitative data from primary research studies that examines the effectiveness of healthcare interventions. To activate evidence-based nursing care in various healthcare settings, the best and available scientific evidence are essential components. This paper will include some examples to promote understandings.


Cortical-layer-specific effects of PACAP and tPA on interneuron migration during post-natal development of the cerebellum.
During early post-natal development of the cerebellum, granule neurons (GN) execute a centripetal migration toward the internal granular layer, whereas basket and stellate cells (B/SC) migrate centrifugally to reach their final position in the molecular layer (ML). We have previously shown that pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates in vitro the expression and release of the serine protease tissue-type plasminogen activator (tPA) from GN, but the coordinated role of PACAP and tPA during interneuron migration has not yet been investigated. Here, we show that endogenous PACAP is responsible for the transient arrest phase of GN at the level of the Purkinje cell layer (PCL) but has no effect on B/SC. tPA is devoid of direct effect on GN motility in vitro, although it is widely distributed along interneuron migratory routes in the ML, PCL, and internal granular layer. Interestingly, plasminogen activator inhibitor 1 reduces the migration speed of GN in the ML and PCL, and that of B/SC in the ML. Taken together, these results reveal for the first time that tPA facilitates the migration of both GN and fast B/SC at the level of their intersection in the ML through degradation of the extracellular matrix. Crucial role of tissue plasminogen activator (tPA) in interneuron migration. Interneuron migration is a critical step for normal establishment of neuronal network. This study indicates that, in the post-natal cerebellum, tPA facilitates the opposite migration of immature excitatory granule neurons (GN) and immature inhibitory basket/stellate cells (B/SC) along the same migratory route. These data show that tPA exerts a pivotal role in neurodevelopment.


Real-Time Bladder Pressure Estimation for Closed-Loop Control in a Detrusor Overactivity Model.
Overactive bladder (OAB) patients suffer from a frequent urge to urinate, which can lead to a poor quality of life. Current neurostimulation therapy uses open-loop electrical stimulation to alleviate symptoms. Continuous stimulation facilitates habituation of neural pathways and consumes battery power. Sensory feedback-based closed-loop stimulation may offer greater clinical benefit by driving bladder relaxation only when bladder contractions are detected, leading to increased bladder capacity. Effective delivery of such sensory feedback, particularly in real-time, is necessary to accomplish this goal. We implemented a Kalman filter-based model to estimate bladder pressure in real-time using unsorted neural recordings from sacral-level dorsal root ganglia, achieving a 0.88 ± 0.16 correlation coefficient fit across 35 normal and simulated OAB bladder fills in five experiments. We also demonstrated closed-loop neuromodulation using the estimated pressure to trigger pudendal nerve stimulation, which increased bladder capacity by 40% in two trials. An offline analysis indicated that unsorted neural signals had a similar stability over time as compared to sorted single units, which would require a higher computational load. We believe this paper demonstrates the utility of decoding bladder pressure from neural activity for closed-loop control; however, real-time validation during behavioral studies is necessary prior to clinical translation.


A global metabolic shift is linked to Salmonella multicellular development.
Bacteria can elaborate complex patterns of development that are dictated by temporally ordered patterns of gene expression, typically under the control of a master regulatory pathway. For some processes, such as biofilm development, regulators that initiate the process have been identified but subsequent phenotypic changes such as stress tolerance do not seem to be under the control of these same regulators. A hallmark feature of biofilms is growth within a self-produced extracellular matrix. In this study we used metabolomics to compare Salmonella cells in rdar colony biofilms to isogenic csgD deletion mutants that do not produce an extracellular matrix. The two populations show distinct metabolite profiles. Even though CsgD controls only extracellular matrix production, metabolite signatures associated with cellular adaptations associated with stress tolerances were present in the wild type but not the mutant cells. To further explore these differences we examine the temporal gene expression of genes implicated in biofilm development and stress adaptations. In wild type cells, genes involved in a metabolic shift to gluconeogenesis and various stress-resistance pathways exhibited an ordered expression profile timed with multicellular development even though they are not CsgD regulated. In csgD mutant cells, the ordered expression was lost. We conclude that the induction of these pathways results from production of, and growth within, a self produced matrix rather than elaboration of a defined genetic program. These results predict that common physiological properties of biofilms are induced independently of regulatory pathways that initiate biofilm formation.


Preterm infants' T cell responses to inactivated poliovirus vaccine.
The antigen-specific T cell responses of preterm infants to immunization are not well understood. The aim of the present study was to compare the T cell responses of preterm infants after inactivated poliovirus vaccination with those of term infants. We prospectively enrolled 2-month-old preterm (gestational age, 33 weeks) and term (gestational age, 37 weeks) infants to receive 3 doses of diphtheria-tetanus toxoids-acellular pertussis-hepatitis B virus-inactivated poliovirus vaccine. Whole blood and peripheral blood mononuclear cells (PBMCs) were stimulated with poliovirus vaccine, and memory T cell activation was analyzed by flow cytometry and lymphoproliferation, respectively. Levels of poliovirus neutralizing antibodies were measured in serum. We enrolled 33 preterm and 50 term infants. Preterm infants had fewer circulating CD4(+)CD45RO(+) memory (P = .005) and CD4(+)CD69(+)IFN-gamma(+) cells activated by staphylococcus enterotoxin B at 2 (P = .015) and 7 (P = .05) months of age. After immunization, preterm and term infants had comparable frequencies of poliovirus-specific CD4(+)CD45RO(+)CD69(+)IFN-gamma(+) memory T cells (P = .79). PBMCs from preterm infants had diminished poliovirus-specific lymphoproliferation (P<.001). Although all infants developed seroprotective poliovirus antibody titers, serotype 1 titers were lower among preterm infants (P = .03). Preterm infants develop poliovirus-specific T cell responses that are comparable to those of term infants. However, they demonstrate nonspecific and poliovirus-specific functional T cell limitations, suggesting that investigations into whether T cell differences remain as preterm infants mature are warranted.


Epinephrine, glucose, and lactate infusion in exercising adrenodemedullated rats.
The purpose of this study was to determine the metabolic function of the marked increase in plasma epinephrine which occurs in fasted rats during treadmill exercise. Fasted adrenodemedullated (ADM) and sham-operated (SHAM) rats were run on a rodent treadmill (21 m/min, 15% grade) for 30 min or until exhaustion. ADM rats were infused with saline, epinephrine, glucose, or lactate during the exercise bouts. ADM saline-infused rats showed markedly reduced endurance, hypoglycemia, elevated plasma insulin, reduced blood lactate, and reduced muscle glycogenolysis compared with exercising SHAM's. Epinephrine infusion corrected all deficiencies. Glucose infusion restored endurance run times and blood glucose to normal without correcting the deficiencies in blood lactate and muscle glycogenolysis. Infusion of lactate partially corrected the hypoglycemia at 30 min of exercise, but endurance was not restored to normal and rats were hypoglycemic at exhaustion. We conclude that in the fasted exercising rat, actions of epinephrine in addition to provision of gluconeogenic substrate are essential for preventing hypoglycemia and allowing the rat to run for long periods of time.


The effect of implementation strength of basic emergency obstetric and newborn care (BEmONC) on facility deliveries and the met need for BEmONC at the primary health care level in Ethiopia.
Basic emergency obstetric and newborn care (BEmONC) is a primary health care level initiative promoted in low- and middle-income countries to reduce maternal and newborn mortality. Tailored support, including BEmONC training to providers, mentoring and monitoring through supportive supervision, provision of equipment and supplies, strengthening referral linkages, and improving infection-prevention practice, was provided in a package of interventions to 134 health centers, covering 91 rural districts of Ethiopia to ensure timely BEmONC care. In recent years, there has been a growing interest in measuring program implementation strength to evaluate public health gains. To assess the effectiveness of the BEmONC initiative, this study measures its implementation strength and examines the effect of its variability across intervention health centers on the rate of facility deliveries and the met need for BEmONC. Before and after data from 134 intervention health centers were collected in April 2013 and July 2015. A BEmONC implementation strength index was constructed from seven input and five process indicators measured through observation, record review, and provider interview; while facility delivery rate and the met need for expected obstetric complications were measured from service statistics and patient records. We estimated the dose-response relationships between outcome and explanatory variables of interest using regression methods. The BEmONC implementation strength index score, which ranged between zero and 10, increased statistically significantly from 4.3 at baseline to 6.7 at follow-up (p < .05). Correspondingly, the health center delivery rate significantly increased from 24% to 56% (p < .05). There was a dose-response relationship between the explanatory and outcome variables. For every unit increase in BEmONC implementation strength score there was a corresponding average of 4.5 percentage points (95% confidence interval: 2.1-6.9) increase in facility-based deliveries; while a higher score for BEmONC implementation strength of a health facility at follow-up was associated with a higher met need. The BEmONC initiative was effective in improving institutional deliveries and may have also improved the met need for BEmONC services. The BEmONC implementation strength index can be potentially used to monitor the implementation of BEmONC interventions.


A third study on predicting NCLEX success with the HESI exit Exam.
This was the third annual validity study designed to assess the accuracy of the HESI Exit Exam (E2) in predicting NCLEX success for graduating registered and practical nursing students. As in year I (N = 2,725) and year II (N = 3,752), in year III (N = 6,277), the E2 was highly predictive of NCLEX success for associate degree nursing, bachelor of science nursing, diploma, and practical nursing students. Unlike previous years, in year IIl, monitoring was not a significant factor in the predictive accuracy of the E2. NCLEX success of low-scoring E2 students, first examined in year II, was also examined in year Ill. As in year II, low-scoring E2 students were significantly more (P = .001) likely to fail the licensure examination than high-scoring E2 students, in year Ill, unlike year II, there was no significant difference in the pass rate of low-scoring E2 students who participated in a remediation program and those who did not. The authors recommended that a more definitive definition of remediation be used in future studies and that such studies focus on E2 implementation strategies and their relationship to NCLEX success.


Methylene blue sensitized dichromated gelatin holograms: a study of their storage life and reprocessibility.
The results of an experimental investigation on the storage life and reprocessibility of methylene blue sensitized dichromated gelatin (MBDCG) holograms are reported. The major conclusions of the investigation are: (i) Storage of MBDCG holograms in normal laboratory conditions for long periods is possible and it diminishes somewhat their diffraction efficiency. (ii) The results on short time storage and long time storage are almost similar, thus indicating that the diffraction efficiency can be stabilized through storage in a relatively short period of time. (iii) The deterioration in the diffraction efficiency on storage is less [D(eta) < 20%] for gratings of low/medium initial efficiency (eta < 70%) and it is more for gratings of high initial efficiency. (iv) About 65-95% restoration of the diffraction efficiency can be accomplished through reprocessing. (v) The restoration of diffraction efficiency is almost perfect [R(eta) > 80%] for gratings of low/medium initial efficiency (eta <75%) whereas it is rather imperfect for gratings having high initial efficiency.


Undetected eye disease in a primary care clinic population.
Population-based data have indicated that a significant proportion of persons with undiagnosed ocular disease in the community are regular users of general medical services. This, combined with the high prevalence of chronic medical disorders known to be risk factors for ocular disease in such clinics, makes them an attractive site for screening. The prevalence of ocular disease was estimated in a sample of 405 general medicine patients attending an adult primary care clinic in an urban teaching hospital. Overall, 205 (50.6%) of 405 patients were found to have clinically important ocular pathology. One third of those affected (n = 68) were unaware of their eye disease, and 26% (n = 18) of these 68 patients required immediate medical or surgical intervention. Patients 65 years or older (odds ratio [OR], 1.76), in fair or poor general health (OR, 1.78), with diabetes mellitus (OR, 2.07), or with self-reported fair or poor vision (OR, 3.03), were at increased risk for the presence of ocular disease. Among patients with eye disease, those who had no insurance coverage for eye care (OR, 3.45), those who had not had an eye examination during the previous 2 years (OR, 4.03), and those whose last eye examination was performed by an optometrist (OR, 7.25, reference ophthalmologist) were more likely to not be aware of their eye disease. Our results underscore the importance of screening for ocular disease in primary health care settings, especially for patients who are older than 65 years, are in poor health, report poor vision, have had infrequent eye examinations, or have inadequate insurance coverage for eye care.


Passive probing of the sound fixing and ranging channel with hydro-acoustic observations from ridge earthquakes.
The International Monitoring System includes a hydro-acoustic part to verify the Comprehensive Nuclear-Test-Ban Treaty. Besides explosive signals, monitoring stations also detect acoustic waves from earthquakes that travel through the SOund Fixing And Ranging (SOFAR) channel. The travel times of such detections are listed in the Reviewed Event Bulletin, which is statistically evaluated for the stations located in the Pacific, Indian, and Atlantic Oceans. The celerities of ridge earthquakes are calculated to build up a homogeneous data-set, based on similar source mechanisms. The celerity is defined as the epicentral distance divided by the travel time. The global characteristics of these celerities can be well understood in terms of temperature variations in the SOFAR channel. A detailed velocity profile has been retrieved for the Atlantic Ocean where large differences (14 m/s) are found between the southern and northern parts of the basin. Propagation modeling with normal modes supports these findings, which shows that the celerity gives an estimate of the sound speed in the SOFAR channel. These results compare remarkably well with those from active experiments, showing the ability of passively probing the SOFAR channel with hydro-acoustic waves from earthquake sources.


Independence between friction and velocity distribution in fluids subjected to severe shearing and confinement.
Friction reduction is more than ever a key point in saving natural resources and energy, and the question of how to achieve this concerns first and foremost every lubricated system. Among the observed phenomena related to lubricated friction, limiting shear stress (LSS) appears to be one of the most challenging to explain, since its origin is still uncertain. Various scenarios have been proposed involving a transition to a glassy state under high pressure, shear banding, shear localization, or even the occurrence of slip at the solid-liquid interface, none of which have proven conclusive. This work provides new insights into the mechanisms leading to LSS and the underlying flow organization. It bridges a gap between the scenarios previously discussed in the literature to explain the mechanisms behind LSS and friction experiments, which provide macroscopic results only. We first present some general molecular dynamics (MD) simulations developed to characterize molecular fluids in their bulk state and then proceed to study their response under severe shearing and confinement. Results from simulations are compared to experimental data derived from friction tests. A further analysis of the pressure and temperature involved hints that LSS and the physical state of the lubricant are strongly interconnected concepts. Additionally, the friction results were uncorrelated to the choice of surfaces, contrary to velocity distribution.


The SET1 Complex Selects Actively Transcribed Target Genes via Multivalent Interaction with CpG Island Chromatin.
Chromatin modifications and the promoter-associated epigenome are important for the regulation of gene expression. However, the mechanisms by which chromatin-modifying complexes are targeted to the appropriate gene promoters in vertebrates and how they influence gene expression have remained poorly defined. Here, using a combination of live-cell imaging and functional genomics, we discover that the vertebrate SET1 complex is targeted to actively transcribed gene promoters through CFP1, which engages in a form of multivalent chromatin reading that involves recognition of non-methylated DNA and histone H3 lysine 4 trimethylation (H3K4me3). CFP1 defines SET1 complex occupancy on chromatin, and its multivalent interactions are required for the SET1 complex to place H3K4me3. In the absence of CFP1, gene expression is perturbed, suggesting that normal targeting and function of the SET1 complex are central to creating an appropriately functioning vertebrate promoter-associated epigenome.


Isolation of clones of rat striatum-specific mRNAs by directional tag PCR subtraction.
We report an improved subtractive cDNA cloning procedure, named "directional tag PCR subtraction," for isolating clones of mRNAs enriched in a target tissue compared to a second tissue, the driver. In this method, the target and driver are prepared from directional cDNA libraries constructed in different vectors, and the target cDNA contains tag sequences at both its 5' and 3' ends for PCR amplification. This method avoids several limitations of previous subtraction procedures, and was demonstrated to be technically easy and efficient. Using the directional tag PCR subtraction and improved screening procedures, cDNA clones corresponding to mRNAs expressed in the striatum but not in the cerebellum of the rat brain were efficiently isolated, including mRNAs encoding calmodulin-dependent phosphodiesterase, a transcriptional regulatory protein, and several previously uncharacterized species. Our data suggest that approximately 1% of the striatal polyA+ RNA mass potentially encoding more than 300 distinct proteins corresponds to RNA species reduced in concentration or absent from the cerebellum, of which about one-third are expressed prominently only in the striatum. This unexpected finding suggests that the striatum has a unique biochemical character within the brain, and that characterization of these mRNAs will be important for understanding the biochemical basis of striatal function.


Voluntary weight reduction in older men increases hip bone loss: the osteoporotic fractures in men study.
To test the hypothesis that weight loss in older men is associated with increased rates of hip bone loss regardless of adiposity and intention to lose weight, we measured body weight, body composition, hip bone mineral density (BMD), and intention to lose weight in a cohort of 1342 older men enrolled in the Osteoporotic Fractures in Men (MrOS) study and followed them prospectively for an average of 1.8 yr for changes in weight and BMD. The adjusted average rate of change in total hip BMD was 0.1%/yr in men with weight gain, -0.3%/yr in men with stable weight, and -1.4%/yr in men with weight loss (test for trend, P < 0.001). Higher rates of hip bone loss were observed in men with weight loss regardless of category of body mass index, body composition, or intention to lose weight. Even among obese (body mass index, > or =30 kg/m2) men trying to lose weight, those with documented voluntary weight reduction experienced an increase in hip bone loss (average rate of change in total hip BMD, 0.5%/yr in those with weight gain, -0.1%/yr in those with stable weight, and -1.7%/yr in those with weight loss; test for trend, P < 0.001). Older men who experience weight loss have increased rates of hip bone loss, even among overweight and obese men undergoing voluntary weight reduction.


The measurement of radiation dose profiles for electron-beam computed tomography using film dosimetry.
The unique geometry of electron-beam CT (EBCT) scanners produces radiation dose profiles with widths which can be considerably different from the corresponding nominal scan width. Additionally, EBCT scanners produce both complex (multiple-slice) and narrow (3 mm) radiation profiles. This work describes the measurement of the axial dose distribution from EBCT within a scattering phantom using film dosimetry methods, which offer increased convenience and spatial resolution compared to thermoluminescent dosimetry (TLD) techniques. Therapy localization film was cut into 8 x 220 mm strips and placed within specially constructed light-tight holders for placement within the cavities of a CT Dose Index (CTDI) phantom. The film was calibrated using a conventional overhead x-ray tube with spectral characteristics matched to the EBCT scanner (130 kVp, 10 mm A1 HVL). The films were digitized at five samples per mm and calibrated dose profiles plotted as a function of z-axis position. Errors due to angle-of-incidence and beam hardening were estimated to be less than 5% and 10%, respectively. The integral exposure under film dose profiles agreed with ion-chamber measurements to within 15%. Exposures measured along the radiation profile differed from TLD measurements by an average of 5%. The film technique provided acceptable accuracy and convenience in comparison to conventional TLD methods, and allowed high spatial-resolution measurement of EBCT radiation dose profiles.


Hydrogen production from inexhaustible supplies of fresh and salt water using microbial reverse-electrodialysis electrolysis cells.
There is a tremendous source of entropic energy available from the salinity difference between river water and seawater, but this energy has yet to be efficiently captured and stored. Here we demonstrate that H(2) can be produced in a single process by capturing the salinity driven energy along with organic matter degradation using exoelectrogenic bacteria. Only five pairs of seawater and river water cells were sandwiched between an anode, containing exoelectrogenic bacteria, and a cathode, forming a microbial reverse-electrodialysis electrolysis cell. Exoelectrogens added an electrical potential from acetate oxidation and reduced the anode overpotential, while the reverse electrodialysis stack contributed 0.5-0.6 V at a salinity ratio (seawater:river water) of 50. The H(2) production rate increased from 0.8 to 1.6 m(3)-H(2)/m(3)-anolyte/day for seawater and river water flow rates ranging from 0.1 to 0.8 mL/ min. H(2) recovery, the ratio of electrons used for H(2) evolution to electrons released by substrate oxidation, ranged from 72% to 86%. Energy efficiencies, calculated from changes in salinities and the loss of organic matter, were 58% to 64%. By using a relatively small reverse electrodialysis stack (11 membranes), only ~1% of the produced energy was needed for pumping water. Although Pt was used on the cathode in these tests, additional tests with a nonprecious metal catalyst (MoS(2)) demonstrated H(2) production at a rate of 0.8 m(3)/m(3)/d and an energy efficiency of 51%. These results show that pure H(2) gas can efficiently be produced from virtually limitless supplies of seawater and river water, and biodegradable organic matter.


Successful Strategies for Engaging Chinese Breast Cancer Survivors in a Randomized Controlled Trial.
Chinese immigrant breast cancer survivors face various challenges due to cultural and socioecological factors. Research efforts to develop culturally sensitive interventions have been limited by lack of knowledge regarding successful recruitment and implementation practices among Chinese immigrant populations. This paper documents strategies utilized during the development and implementation of a randomized controlled trial of a culturally sensitive psychosocial intervention for Chinese immigrant breast cancer survivors. In partnership with a community agency, we developed culturally and linguistically appropriate research materials, recruited participants from community channels, and conducted longitudinal data collection. Key strategies include building equitable research partnerships with community agencies to engage participants; being responsive to the needs of community agencies and participants; considering within-group diversity of the research population; utilizing recruitment as an opportunity for relationship-building with participants; and developing key strategies to promote retention. Successful participant engagement in cancer intervention research is the result of collaboration among breast cancer survivors, community leaders and agencies, and academic researchers. The engagement process for this study is novel because we have emphasized cultural factors in the process and taken a relational approach to recruitment and retention.


Tube-like structures with co-expression of D2-40 and CD34: newly formed vasculatures?
A great number of in vitro and in vivo studies have suggested that many pathways or factors can stimulate angiogenesis and lymphangiogenesis, which facilitate tumor progression and metastasis. However, the morphological and immunohistochemical profile of newly formed vasculatures has not been elucidated, making it difficult to differentiate them from the pre-existing ones, and to identify their unique molecular profiles for diagnosis and therapeutic interventions. As cytokeratin (CK)-19 is a well-recognized stem cell marker and CK-19-positive cells are frequently detected in the peripheral blood of patients with metastatic cancer, our recent studies have assessed the involvement of CK-19 in the formation of new vasculatures in primary colorectal cancer (CRC) tissues. Our studies showed that a subset of lymph node-positive cases harbored some isolated normal epithelial structures with distinct CK-19 immunostaining within an otherwise CK-19-negative background. These structures are exclusively located within or adjacent to lymphoid follicles and are often surrounded by tube-like structures expressing lymphatic endothelial marker D2-40. Similar structures are more frequently seen at the junctions between pre-invasive and invasive CRC with the following features: (1). they consist of a single layer of endothelial cells that express both D2-40 and CD34, (2). their endothelial walls are often incomplete with disseminated cells protruding into the adjacent stroma, and (3). they are exclusively associated with disseminated CK-19-positive cells Based on these findings, we propose that these tube-like structures represent newly formed vasculatures, which are derived by the convergence of aberrant lymphocyte infiltration and tumor stem cells. Because of their close physical proximity, tumor stem cells within the epithelial and stromal components contribute equally and coordinately to the morphogenesis of new vasculatures, which constitutes the basis for the unique morphologic and immunohistochemical features of newly formed vasculatures. Our hypothesis appears to be applicable to all epithelium-derived cancers.


Three dimensional optical angiography.
With existing optical imaging techniques three-dimensional (3-D) mapping of microvascular perfusion within tissue beds is severely limited by the efficient scattering and absorption of light by tissue. To overcome these limitations we have developed a method of optical angiography (OAG) that can generate 3-D angiograms within millimeter tissue depths by analyzing the endogenous optical scattering signal from an illuminated sample. The technique effectively separates the moving and static scattering elements within tissue to achieve high resolution images of blood flow, mapped into the 3-D optically sectioned tissue beds, at speeds that allow for perfusion assessment in vivo. Its development has its origin in Fourier domain optical coherence tomography. We used OAG to visualize the cerebral microcirculation, of adult living mice through the intact cranium, measurements which would be difficult, if not impossible, with other optical imaging techniques.


Optimum spatiotemporal receptive fields for vision in dim light.
Many nocturnal insects depend on vision for daily life and have evolved different strategies to improve their visual capabilities in dim light. Neural summation of visual signals is one strategy to improve visual performance, and this is likely to be especially important for insects with apposition compound eyes. Here we develop a model to determine the optimum spatiotemporal sampling of natural scenes at gradually decreasing light levels. Image anisotropy has a strong influence on the receptive field properties predicted to be optimal at low light intensities. Spatial summation between visual channels is predicted to extend more strongly in the direction with higher correlations between the input signals. Increased spatiotemporal summation increases signal-to-noise ratio at low frequencies but sacrifices signal-to-noise ratio at higher frequencies. These results, while obtained from a model of the insect visual system, are likely to apply to visual systems in general.


Systematic comparison of sporadic and syndromic pancreatic islet cell tumors.
Pancreatic islet cell tumors (ICTs) occur as sporadic neoplasias or as a manifestation of multiple endocrine neoplasia type 1 (MEN1) and von Hippel-Lindau disease (VHL). Molecular classification of ICTs is mandatory for timely diagnosis and surveillance. Systematic comparison of VHL-ICTs and sporadic ICTs has been lacking. Our registry-based approaches used the German NET-Registry with 259 patients with neuroendocrine tumors (NETs), who were primarily diagnosed with NETs, and the German VHL-Registry with 485 molecular genetically confirmed patients who had undergone magnetic resonance imaging or computed tomography of the abdomen. All patients provided blood DNA for testing of the MEN1 and VHL genes for intragenic mutations and large deletions. In the NET-Registry, 9/101 patients (8.9%) with ICTs had germline mutations, 8 in MEN1 and 1 in VHL. In the VHL-Registry, prevalence of NETs was 52/487 (10.6%), and all were ICTs. Interestingly, of those with VHL p.R167W, 47% developed ICTs, compared to 2% of those with p.Y98H. In total, there were 92 truly sporadic, i.e. mutation-negative ICT patients. Comparing these with the 53 VHL-ICT patients, the statistically significant differences were predominance of female gender (P=0.01), multifocal ICTs (P=0.0029), and lower malignancy rate (P<0.001) in VHL-ICTs compared to sporadic cases. VHL was prevalent in <0.5% of NETs, while NETs occur in ∼10% of VHL, virtually exclusively as ICTs, which are rarely the first presentation. Patients with NETs should not be subjected to genetic testing of the VHL gene, unless they have multifocal ICTs, other VHL-associated tumors, and/or a family history for VHL.