Tardigrades exhibit robust interlimb coordination across walking speeds and terrains.
Tardigrades must negotiate heterogeneous, fluctuating environments and accordingly utilize locomotive strategies capable of dealing with variable terrain. We analyze the kinematics and interleg coordination of freely walking tardigrades (species: Hypsibius exemplaris). We find that tardigrade walking replicates several key features of walking in insects despite disparities in size, skeleton, and habitat. To test the effect of environmental changes on tardigrade locomotor control circuits we measure kinematics and interleg coordination during walking on two substrates of different stiffnesses. We find that the phase offset between contralateral leg pairs is flexible, while ipsilateral coordination is preserved across environmental conditions. This mirrors similar results in insects and crustaceans. We propose that these functional similarities in walking coordination between tardigrades and arthropods is either due to a generalized locomotor control circuit common to panarthropods or to independent convergence onto an optimal strategy for robust multilegged control in small animals with simple circuitry. Our results highlight the value of tardigrades as a comparative system toward understanding the mechanisms-neural and/or mechanical-underlying coordination in panarthropod locomotion.


Assessing utilization of a marketwide price transparency tool.
Most transparency tools are provided by individual insurers for enrollees shopping for services within their networks. This paper seeks to understand the impact of a marketwide price transparency tool with an embedded randomized experiment to offer provider-level charge information. In September 2017, FAIR Health released an independent, publicly accessible statewide consumer shopping tool, New York Healthcare Online Shopping Tool, or NYHOST, that displays individual provider charges (list prices) for common procedures in each of New York State's 3-digit geozips, as well as the estimated insurer-allowed amounts and educational resources. The rollout was accompanied by an extensive, multipronged marketing effort. It also incorporated a randomized experiment: The set of procedures with provider-level information varied across areas. We characterized the types of services that were most searched on the consumer shopping tool. Utilizing negative binomial models with procedure and area fixed effects, we report on market and procedural characteristics that influence price search. Consumers utilized the tool strategically, searching more in procedure markets with provider-specific price information availability, more frequent out-of-network utilization, higher charges, significant charge dispersion, and substantial provider competition. We also found that the majority of searches using the tool were for emergent services not usually classified as shoppable, suggesting that consumers may also have used the tool to compare searchable prices against those in bills for services already received. Our findings confirm aspects of price search theory that have been developed by studying other industries and may prove instructive for further study of price transparency tools.


Modulating birth weight heritability in mice.
Expected genetic response is proportional to the heritability of the trait, and this parameter is considered inherent of a specific trait in a particular population. However, models assuming heterogeneity in residual variance lead to different estimates of heritability across combinations of systematic (environmental) effects. Modifying the residual variance of the birth weight by artificial selection was shown to be feasible in a divergent selection experiment in mice. The objectives of this work were to 1) estimate the evolution of the heritability of birth weight in mice in the mentioned experiment, and 2) estimate different heritability regarding systematic effects. Data came from eleven generations of a divergent selection experiment to modify the residual variability of birth weight in mice. A total of 15,431 birth weight records from 959 females and 1,641 litters in combination with 14,786 pedigree records were used. The model used for analysis included generation, litter size, sex, and parity number as systematic effects. Each record of birth weight was assigned to the mother of the pup in the model which assumes that the residual variance is heterogeneous and partially under genetic control. Differences in heritability between lines reached values of 0.06 in the last generations. Choosing the most extreme values of systematic effects, the birth weight heritability ranged from 0.04 to 0.22. From these results, the possibility of modulating the heritability for this trait could be explored in 1 of 2 ways: selecting to decrease the residual variability, or choosing the specific levels of the systematic effects.


A common methodological phylogenomics framework for intra-patient heteroplasmies to infer SARS-CoV-2 sublineages and tumor clones.
All diseases containing genetic material undergo genetic evolution and give rise to heterogeneity including cancer and infection. Although these illnesses are biologically very different, the ability for phylogenetic retrodiction based on the genomic reads is common between them and thus tree-based principles and assumptions are shared. Just as the different frequencies of tumor genomic variants presupposes the existence of multiple tumor clones and provides a handle to computationally infer them, we postulate that the different variant frequencies in viral reads offers the means to infer multiple co-infecting sublineages. We present a common methodological framework to infer the phylogenomics from genomic data, be it reads of SARS-CoV-2 of multiple COVID-19 patients or bulk DNAseq of the tumor of a cancer patient. We describe the Concerti computational framework for inferring phylogenies in each of the two scenarios.To demonstrate the accuracy of the method, we reproduce some known results in both scenarios. We also make some additional discoveries. Concerti successfully extracts and integrates information from multi-point samples, enabling the discovery of clinically plausible phylogenetic trees that capture the heterogeneity known to exist both spatially and temporally. These models can have direct therapeutic implications by highlighting "birth" of clones that may harbor resistance mechanisms to treatment, "death" of subclones with drug targets, and acquisition of functionally pertinent mutations in clones that may have seemed clinically irrelevant. Specifically in this paper we uncover new potential parallel mutations in the evolution of the SARS-CoV-2 virus. In the context of cancer, we identify new clones harboring resistant mutations to therapy.


Signal detection theory approach to gastroesophageal reflux disease: a new method for symptom analysis of impedance-pH data.
An accurate reflux-symptom relationship analysis method is an unmet need in gastroesophageal reflux disease (GERD) diagnosis. The aim of this study was to adapt signal detection theory (SDT) approach to reflux-symptom relationship analysis to develop a new diagnosis method. Patients with predominant symptoms of heartburn and regurgitation were enrolled. Proton pump inhibitor (PPI)-responsive and PPI-unresponsive groups were created via interview and PPI trial. Patients then underwent stationary esophageal manometry and 24-hour multichannel intraluminal impedance-pH monitoring. SDT measurement parameters (discriminability: d' and criterion: c) were calculated using empirically selected time windows (0.5, 1, 2, 3, 4 and 5 minutes). The time window that provided the highest d' value was selected as the optimal time window. A cut-off d' value that optimally separates two groups was found using receiver operating characteristics analysis. Sixty-three patients completed the study (45 PPI responsive). Optimal time window and cut-off d' value were found as 1 and 0.767 minute, respectively. Symptom association probability (SAP) index values showed good correlation (rS = 0.7182, P < 0.0001) with d' values. SDT approach to reflux-symptom relationship analysis showed sensitivity (89% vs. 78%) and negative predictive values (75% vs. 60%) favorable over SAP index analysis. SDT approach using 1-minute time window and 0.767 cut-off d' value provides us a new and more accurate measure of reflux-symptom relationship than SAP index analysis.


A double blind trial of extra-amniotic oestriol and prostaglandin F2 alpha gels in cervical ripening.
Forty nulliparous patients with Bishop score of 3 or less were given 15 mg oestriol gel, 10 mg PGF2 alpha gel or gel alone via the extra-amniotic route. Both the oestriol and prostaglandin treated patients had a significant increase in Bishop score and a significant reduction in induction-delivery interval compared to controls. Oestriol gel had significantly less stimulatory effect on uterine activity than prostaglandin gel, indicating a possible local action.


Plasma Thermogram Parameters Differentiate Status and Overall Survival of Melanoma Patients.
Melanoma is the fifth most common cancer in the United States and the deadliest of all skin cancers. Even with recent advancements in treatment, there is still a 13% two-year recurrence rate, with approximately 30% of recurrences being distant metastases. Identifying patients at high risk for recurrence or advanced disease is critical for optimal clinical decision-making. Currently, there is substantial variability in the selection of screening tests and imaging, with most modalities characterized by relatively low accuracy. In the current study, we built upon a preliminary examination of differential scanning calorimetry (DSC) in the melanoma setting to examine its utility for diagnostic and prognostic assessment. Using regression analysis, we found that selected DSC profile (thermogram) parameters were useful for differentiation between melanoma patients and healthy controls, with more complex models distinguishing melanoma patients with no evidence of disease from patients with active disease. Thermogram features contributing to the third principal component (PC3) were useful for differentiation between controls and melanoma patients, and Cox proportional hazards regression analysis indicated that PC3 was useful for predicting the overall survival of active melanoma patients. With the further development and optimization of the classification method, DSC could complement current diagnostic strategies to improve screening, diagnosis, and prognosis of melanoma patients.


Clinical value of mammography in diagnosis and identification of breast mass.
To study the effect and clinical value of mammography in the diagnosis of breast lump so as to improve the diagnosis level of breast cancer. A retrospective analysis was carried out on clinical data of 110 patients with mammary lump confirmed by pathology to study the compliance of mammography diagnosis and Pathology diagnosis in breast lump, and the detection of microcalcifications, phyllode, and observe the image performance of mammography. Taking infitrating ductal carcinoma (IDC) as an example, the correlation of image performance and clinical pathological features of different types was studied so as to predict if mammography performance was effective in the treatment and prognosis in breast cancer. Taking Breast Imaging Reporting and Data System (BI-RADS) grade 4A as the critical point, the sensitivity, specificity and accuracy of mammography was 90.80% (109/120), 84.60% (126/149) and 87.40% (235/269); taking BI-RADS grade 4B as the critical point, the sensitivity, specificity and accuracy of mammography was 85.00% (102/120), 93.30% (139/149) and 89.60% (241/269); the correlation analysis suggested that, there was some kind of correlation between the mammography performance and clinical features of breast cancer. Mammography is worth being promoted in clinic for its significant clinical value in diagnosing and identifying breast lump.


Chaintweak: sampling from the neighbourhood of a protein conformation.
When searching for an optimal protein structure, it is often necessary to generate a set of structures similar, e.g., within 4A Root Mean Square Deviation (RMSD), to some base structure. Current methods to do this are designed to produce only small deviations (< 0.1A RMSD) and are inefficient for larger deviations. The method proposed in this paper, ChainTweak, can generate conformations with larger deviations from the base much more efficiently. For example, in 18 seconds it can generate 100 backbone conformations, each within 1-4A RMSD of a given 45-residue conformation. Moreover, each conformation has correct bond lengths, angles and omega torsional angles; its phi-psi angles have energetically favorable values; and there are rarely any backbone steric clashes. The method uses the insight that loop closure techniques can be used to perform compensatory changes of dihedral angles so that only a part of the conformation is changed. It is demonstrated, using decoys from the Decoys 'R Us data-set, that ChainTweak can be used to construct good decoys. It also provides a novel and intuitive way of analyzing the energy landscape of a protein. In addition, ChainTweak can improve the accuracy and performance of the loop modeling program RAPPER by an order of magnitude (1.1 min. vs. 36 min. for an 8-residue chain).


Aortic arch variant with a common origin of the innominate and left carotid artery as a determinant of thoracic aortic disease: a systematic review and meta-analysis.
The aim of this study was to investigate whether the 'bovine' arch [i.e. arch variant with a common origin of the innominate and left carotid artery (CILCA)] is associated with an increased risk of thoracic aortic disease (TAD). The study was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The PubMed, EMBASE and Cochrane databases were searched to identify all case series reporting about CILCA arch and TAD between January 2008 and December 2018. A total of 485 studies were screened. The prevalence of CILCA arch was assessed, and data analysis was performed considering the difference in the risk of TAD for presence versus absence of CILCA arch. Eight studies enrolling 11 381 subjects were retrieved for quantitative analysis. The proportion of TAD among CILCA arch patients was higher [41.5% (28.1-56.4)] than the proportion among patients with standard arch configuration 34.0% (20.1-51.4). The odds ratio of developing TAD was 1.4 times higher in subjects with CILCA arch (95% confidence interval 1.068-1.839). The test for an overall effect indicated a significant association between CILCA arch and TAD (P < 0.015). The I2 was 78.1% with a value of P < 0.001 for heterogeneity. The Egger test did not show evidence of publication bias (P = 0.317). In conclusion, our meta-analysis supports the hypothesis of a correlation between the presence of CILCA arch and the onset of TAD. Our results warrant a specific and long-term surveillance for patients with this anatomical variant, and a thorough awareness of its potential clinical implications during image interpretation.


Fatalities from road traffic accidents among the young in Bahrain.
In Bahrain and other Gulf countries, road traffic accidents (RTA) are recognized as a growing public health problem. This study described the trend of fatalities from RTA in Bahrain among those aged < 25 years from 2003-2010. The proportional mortality ratios of RTA deaths (up to 30 days from the accident) among the young were compared with those of all ages. The average proportion of young males killed by RTA in Bahrain during the 8-year period was 82.7%, with 2006 marking the lowest proportion among Bahraini nationals (72.2%) and the total population (79.2%). RTA fatalities constituted 51.3% among the Bahrainis, 20.2% of non-Bahrainis and 39.0% of all fatalities in both sexes combined. The young in Bahrain were 3.5 times more likely to die from RTA than the general population. This paper calls on policy-makers to consolidate efforts to control this public health problem.


NY-ESO-1 expression in meningioma suggests a rationale for new immunotherapeutic approaches.
Meningiomas are the most common primary intracranial tumors. Surgical resection remains the treatment of choice for these tumors. However, a significant number of tumors are not surgically accessible, recur, or become malignant, necessitating the repetition of surgery and sometimes radiation. Chemotherapy is rarely used and is generally not recognized as an effective treatment. Cancer/testis (CT) genes represent a unique class of genes, which are expressed by germ cells, normally silenced in somatic cells, but activated in various cancers. CT proteins can elicit spontaneous immune responses in patients with cancer and this feature makes them attractive targets for immunotherapy-based approaches. We analyzed mRNA expression of 37 testis-restricted CT genes in a discovery set of 18 meningiomas by reverse transcription PCR. The overall frequency of expression of CT genes ranged from 5.6% to 27.8%. The most frequently expressed was NY-ESO-1, in 5 patients (27.8%). We subsequently analyzed NY-ESO-1 protein expression in a larger set of meningiomas by immunohistochemistry and found expression in 108 of 110 cases. In some cases, NY-ESO-1 expression was diffused and homogenous, but in most instances it was heterogeneous. Importantly, NY-ESO-1 expression was positively correlated with higher grade and patients presenting with higher levels of NY-ESO-1 staining had significantly worse disease-free and overall survival. We have also shown that NY-ESO-1 expression may lead to humoral immune response in patients with meningioma. Considering the limited treatment options for patients with meningioma, the potential of NY-ESO-1-based immunotherapy should be explored.


An Infratemporal Meningioma: A Diagnostic Dilemma.
A 46-year-old male presented with painless, recurrent bilateral ear discharge and an enlarging right temporal swelling. There were no neurological deficits. Imaging revealed an enhancing, soft tissue mass at the right infratemporal region involving the right temporalis muscle with a small, enhancing intradural component and associated hyperostosis of the greater wing of the right sphenoid bone. Tumour debulking of the right temporalis tumour was performed. Tumour invasion of the right temporalis muscle was noted intraoperatively. Histopathological result was consistent with fibrous meningioma WHO Grade 1 involving surgical resection margins. Follow-up MRI revealed residual right temporal extracranial component. Thus, plans were made for a second stage tumour debulking, however at time of writing, surgery had not been performed. This case highlights the differing appearances of the common meningioma occurring extracranially with elaboration of its differential diagnosis and management.


Identification and verification of novel ferroptosis biomarkers predicts the prognosis of hepatocellular carcinoma.
Big data mining and experiments are widely used to mine new prognostic markers. Candidate genes were identified from CROEMINE and FerrDb. Kaplan-Meier survival and Cox regression analysis were applied to assess the association of genes with Overall survival time (OS) and Disease-free survival time (DFS) in two HCC cohorts. Real-time quantitative polymerase chain reaction (RT-qPCR) and Immunohistochemistry were performed in HCC samples. 21 and 15 genes that can predict OS and DFS, which had not been reported before, were identified from 719 genes, respectively. Survival analysis showed elevated mRNA expression of GLMP, SLC38A6, and WDR76 were associated with poor prognosis, and three genes combination signature was an independent prognostic factor in HCC. RT-qPCR and Immunohistochemistry confirmed the results. We established a novel computational process, which identified the expression levels of GLMP, SLC38A6, and WDR76 as potential ferroptosis-related biomarkers indicating the prognosis of HCC.


The existence and abundance of ghost ancestors in biparental populations.
In a randomly-mating biparental population of size N there are, with high probability, individuals who are genealogical ancestors of every extant individual within approximately log2(N) generations into the past. We use this result of J. Chang to prove a curious corollary under standard models of recombination: there exist, with high probability, individuals within a constant multiple of log2(N) generations into the past who are simultaneously (i) genealogical ancestors of each of the individuals at the present, and (ii) genetic ancestors to none of the individuals at the present. Such ancestral individuals-ancestors of everyone today that left no genetic trace-represent 'ghost' ancestors in a strong sense. In this short note, we use simple analytical argument and simulations to estimate how many such individuals exist in finite Wright-Fisher populations.


Seeing through the stratum corneum.
The stratum corneum (SC) provides a vital barrier membrane between the external environment and the vulnerable internal tissues of the skin. It impedes the flow of water, the penetration of xenobiotics, and invasion of pathogenic micro-organisms. It also has protective capacity against ultraviolet radiation and thermal injury. As routine histopathology provides a misleading picture of a disorganized and shadowy SC, we would recommend the skin surface biopsy technique. This painless technique is easy and reliable in obtaining information from the SC. It demonstrates the geometric patterns of the surface, the openings of the eccrine ducts and hair follicles. The skin surface biopsy technique is also ideal for the investigation of the in situ microbiology of skin. Staining with periodic acid Schiff reagent makes it possible to see ringworm fungi, pityriasis versicolor, candida species, or erythrasma micro-organisms. Scanning electron microscopy can be employed when the higher magnification is needed. Histochemical applications include silver staining for melanin particle, potassium ferricyanide staining for blood pigments and lipid staining with Sudan red, for sebum. The rate of movement of topically applied drugs into the skin can be measured using the skin surface biopsy technique. The concentration of radiolabelled drugs can be counted and compared. Comedogenicity and DNA analysis are other applications of this non-invasive technique.


Single-cell profiling reveals GPCR heterogeneity and functional patterning during neuroinflammation.
GPCR expression was intensively studied in bulk cDNA of leukocyte populations, but limited data are available with respect to expression in individual cells. Here, we show a microfluidic-based single-cell GPCR expression analysis in primary T cells, myeloid cells, and endothelial cells under naive conditions and during experimental autoimmune encephalomyelitis, the mouse model of multiple sclerosis. We found that neuroinflammation induces characteristic changes in GPCR heterogeneity and patterning, and we identify various functionally relevant subgroups with specific GPCR profiles among spinal cord-infiltrating CD4 T cells, macrophages, microglia, or endothelial cells. Using GPCRs CXCR4, S1P1, and LPHN2 as examples, we show how this information can be used to develop new strategies for the functional modulation of Th17 cells and activated endothelial cells. Taken together, single-cell GPCR expression analysis identifies functionally relevant subpopulations with specific GPCR repertoires and provides a basis for the development of new therapeutic strategies in immune disorders.


Generation of thermostable Moloney murine leukemia virus reverse transcriptase variants using site saturation mutagenesis library and cell-free protein expression system.
We attempted to increase the thermostability of Moloney murine leukemia virus (MMLV) reverse transcriptase (RT). The eight-site saturation mutagenesis libraries corresponding to Ala70-Arg469 in the whole MMLV RT (Thr24-Leu671), in each of which 1 out of 50 amino acid residues was replaced with other amino acid residue, were constructed. Seven-hundred and sixty eight MMLV RT clones were expressed using a cell-free protein expression system, and their thermostabilities were assessed by the temperature of thermal treatment at which they retained cDNA synthesis activity. One clone D200C was selected as the most thermostable variant. The highest temperature of thermal treatment at which D200C exhibited cDNA synthesis activity was 57ºC, which was higher than for WT (53ºC). Our results suggest that a combination of site saturation mutagenesis library and cell-free protein expression system might be useful for generation of thermostable MMLV RT in a short period of time for expression and selection.


Proteus syndrome.
The term Proteus syndrome was coined in 1983 to describe a disorder of skeletal, hamartomatous, and other mesodermal malformations. The syndrome was named after the Greek god Proteus, whose name means "the Polymorphous." Clinical features of this new syndrome are currently being defined. Including the case reported herein, we have found 34 patients with Proteus syndrome described in the English literature. Major clinical findings, defined as those findings seen in more than half of the cases, include hemihypertrophy, macrodactyly, exostoses, epidermal nevi, characteristic cerebriform masses involving the plantar or palmar surfaces, a variety of subcutaneous masses, and scoliosis. Histologic examination of subcutaneous masses has identified a variety of lipomatous, hamartomatous, and angiomatous tumors.


Effective and durable genetic modification of human mesenchymal stem cells via controlled release of rAAV vectors from self-assembling peptide hydrogels with a maintained differentiation potency.
Controlling the release of recombinant adeno-associated virus (rAAV) vectors from biocompatible materials is a novel, attractive approach to increase the residence time and effectiveness of a gene carrier at a defined target site. Self-assembling peptides have an ability to form stable hydrogels and encapsulate cells upon exposure to physiological pH and ionic strength. Here, we examined the capacity of the peptide hydrogel RAD16-I in a pure (RAD) form or combined with hyaluronic acid (RAD-HA) to release rAAV vectors as a means to genetically modify primary human bone marrow-derived mesenchymal stem cells (hMSCs), a potent source of cells for regenerative medicine. Specifically, we demonstrate the ability of the systems to efficiently encapsulate and release rAAV vectors in a sustained, controlled manner for the effective transduction of hMSCs (up to 80%) without deleterious effects on cell viability (up to 100%) or on their potential for chondrogenic differentiation over time (up to 21days). The present study demonstrates that RAD16-I is an advantageous material with tunable properties to control the release of rAAV vectors as a promising tool to develop new, improved therapeutic approaches for tissue engineering in vivo.


A phylogenomic approach to bacterial phylogeny: evidence of a core of genes sharing a common history.
It has been claimed that complete genome sequences would clarify phylogenetic relationships between organisms, but up to now, no satisfying approach has been proposed to use efficiently these data. For instance, if the coding of presence or absence of genes in complete genomes gives interesting results, it does not take into account the phylogenetic information contained in sequences and ignores hidden paralogies by using a BLAST reciprocal best hit definition of orthology. In addition, concatenation of sequences of different genes as well as building of consensus trees only consider the few genes that are shared among all organisms. Here we present an attempt to use a supertree method to build the phylogenetic tree of 45 organisms, with special focus on bacterial phylogeny. This led us to perform a phylogenetic study of congruence of tree topologies, which allows the identification of a core of genes supporting similar species phylogeny. We then used this core of genes to infer a tree. This phylogeny presents several differences with the rRNA phylogeny, notably for the position of hyperthermophilic bacteria.


Hormonal regulation of the adrenocortical cell.
Monolayer cultures of bovine and human adrenocortical cells have been used to study regulation of growth and function. Homogeneous bovine adrenocortical cells exhibit a finite life span of approximately 60 generations in culture. Full maintenance of differentiated function (steroid hormone synthesis) requires an inducer such as ACTH and antioxidizing conditions. Full induction of differentiated function occurs only when cellular hypertrophy is stimulated by growth factors such as fibroblast growth factor and serum. ACTH and other agents that increase cellular cAMP inhibit replication but do not block growth factor-induced cellular hypertrophy. ACTH and growth factors together result in a hypertrophied, hyperfunctional cell. Replication ensues only when desensitization to the growth inhibitory effects of ACTH occurs. Cultures of the definitive and fetal zones of the human fetal adrenal cortex synthesize the steroids characteristic of the two zones in vivo. ACTH stimulates production of dehydroepiandrosterone (DHA), the major steroid product of the fetal zone, and of cortisol, the characteristic steroid product of the definitive zone. Prolonged ACTH treatment of fetal zone cultures results in a preferential increase in cortisol production so that the pattern of steroid synthesis becomes that of the definitive zone. The preferential increase in cortisol production by fetal Zone cultures results from induction of 3 beta-hydroxysteroid dehydrogenase, delta 4,5 isomerase activity, which is limiting in fetal zone cells. ACTH thus causes a phenotypic change in fetal zone cells to that of definitive zone cells. In both bovine and human adrenocortical cells, the principal effect of ACTH is to induce full expression of differentiated function. This occurs only under conditions where growth substances and nutrients permit full amplification.


Cardiovascular and renal safety of metformin in patients with diabetes and moderate or severe chronic kidney disease: Observations from the EXSCEL and SAVOR-TIMI 53 cardiovascular outcomes trials.
To provide evidence on the cardiovascular and renal safety of metformin in chronic kidney disease (CKD) stages 3 to 4. This post hoc analysis compared participants with an estimated glomerular filtration rate (eGFR) of 15 to 59 mL/min/1.73m2 in the Exenatide Study of Cardiovascular Event Lowering (EXSCEL) and the Saxagliptin and Cardiovascular Outcomes in Patients With Type 2 Diabetes Mellitus (SAVOR-TIMI 53) trials taking metformin, with those not exposed to metformin during these trials, using a propensity-matching approach. Adjusted Cox proportional hazards models were used to assess risk of major adverse cardiovascular events (MACE) and all-cause mortality (ACM). Metformin effect on eGFR slope was calculated using a mixed-model repeated measures analysis, and the number of lactic acidosis events was tabulated. No strong trend for lower metformin doses with lower eGFR values was observed in either the EXSCEL or SAVOR-TIMI 53 trials. In the 1745 metformin-using participants matched to non-metformin users, metformin had neutral effects on MACE (hazard ratio [HR] 0.91, 95% confidence interval [CI] 0.76-1.08; P = 0.28) and ACM (HR 0.86, 95% CI 0.70-1.07; P = 0.18), with no interaction by CKD stage, or with use of exenatide or saxagliptin. An improvement in eGFR slope was observed with metformin in the CKD stage 3B cohort in SAVOR-TIMI 53, but not in other groups. This analysis of participants with CKD stages 3 to 4 from two cardiovascular outcomes trials supports the cardiorenal safety of metformin, but does not suggest a consistent benefit on MACE, ACM, or eGFR slope across this population.


[Recent advances in the diagnosis and treatment of colorectal cancers].
We review recent developments in the diagnosis and treatment of colorectal cancer. In diagnosis, the detection of the APC gene mutation in stool samples has had a strong impact because of its practical usage for mass screening. Several reports have indicated that the molecular detection of a very small number of cancer cells in the peripheral blood, bone marrow, or lymph nodes yields adequate information on recurrent disease. Several new markers were reported as a significant indicators of cancer development, invasion, or metastasis. In treatment, determination of microsatellite instability may be useful to select good candidates for 5-fluorouracil-based chemotherapy. New molecular targets have been reported, including epidermal growth factor receptor inhibitor, matrix metalloproteinase inhibitors, etc. Gene therapy with p53 or FHIT had undergone successful clinical trials. Tumor-specific immunotherapy with dendritic cell vaccination has also been achieved. In conclusion, many efforts to overcome colorectal cancer will soon open new therapeutic windows.


A Review of Corneal Collagen Cross-linking - Current Trends in Practice Applications.
To review the literature on current applications of corneal Collagen Cross-Linking (CXL). A review of publications on corneal cross-linking was conducted. This included systemic reviews, randomized controlled clinical trials, cohort studies, case-controlled studies and case series. A summary of the publications is tabulated. The original indication of riboflavin - Ultraviolet-A (UVA) induced corneal collagen cross-linking is to arrest the progression of keratoconus. Studies show that it is effective in arresting the progression of keratoconus and post-LASIK ectasia with the standard Dresden protocol (epithelium-off). There are also improvements in visual, keratometric and topographic measurements over time. Severe complications of cross-linking are rare. The epithelium-on techniques have less efficacy than the Dresden protocol. Accelerated protocols have variable results, with some studies reporting comparable outcomes to the Dresden protocol while other studies reporting less efficacious outcomes. Cross-linking combined with refractive procedures provide better visual outcome but long term studies are warranted. Cross-linking for the treatment of infective keratitis is a promising new treatment modality. Initial studies show that it is more effective for superficial rather than deep infections and for bacterial rather than fungal infections. Corneal cross-linking is a procedure with an expanding list of indications from the treatment of corneal ectasias to infective keratitis. While the standard Dresden protocol is established as the gold standard treatment for progressive keratoconus, the more recent protocols may require further refinements, investigative and long-term studies.


Wear testing of a canine hip resurfacing implant that uses highly cross-linked polyethylene.
Hip resurfacing offers advantages for young, active patients afflicted with hip osteoarthritis and may also be a beneficial treatment for adult canines. Conventional hip resurfacing uses metal-on-metal bearings to preserve bone stock, but it may be feasible to use metal-on-polyethylene bearings to reduce metal wear debris while still preserving bone. This study characterized the short-term wear behavior of a novel hip resurfacing implant for canines that uses a 1.5 mm thick liner of highly cross-linked polyethylene in the acetabular component. This implant was tested in an orbital bearing machine that simulated canine gait for 1.1 million cycles. Wear of the liner was evaluated using gravimetric analysis and by measuring wear depth with an optical scanner. The liners had a steady-state mass wear rate of 0.99 ± 0.17 mg per million cycles and an average wear depth in the central liner region of 0.028 mm. No liners, shells, or femoral heads had any catastrophic failure due to yielding or fracture. These results suggest that the thin liners will not prematurely crack after implantation in canines. This is the first hip resurfacing device developed for canines, and this study is the first to characterize the in vitro wear of highly cross-linked polyethylene liners in a hip resurfacing implant. The canine implant developed in this study may be an attractive treatment option for canines afflicted with hip osteoarthritis, and since canines are the preferred animal model for human hip replacement, this implant can support the development of metal-on-polyethylene hip resurfacing technology for human patients. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1196-1205, 2018.


Common Sampling Errors in Research Studies.
Proper sample selection is based on the study purpose, research question(s), and study design. Investigators must use care to select a sample population that is representative of the source and target populations. Well-defined inclusion and exclusion criteria serve as guidance when screening potential candidates for eligibility for participation in a study. Sampling and non-sampling errors may influence study outcomes and generalizability of results. The purpose of this short report is to review common sampling errors made when designing a study and when reporting study outcomes.


[The value of radiotherapy after modified radical mastectomy. 10-year results of the Heidelberg study].
Modified radical mastectomy was performed in 142 female patients with operable carcinoma of the breast. One of the two prospectively randomised groups received postoperative radiotherapy with 6000 rad. Survival rates were equal for both groups. Follow-up radiotherapy had a beneficial influence on survival rate and rate of distant metastases in median tumour sites. In lateral tumour localization the course of disease was less favourable with radiotherapy than without. Local relapse rates were slightly, however not significantly, lower after irradiation. The decisive prognostic criterium is T- and N-stage at the time of diagnosis.


Types of 'reticulin' antibodies detected in human sera by immunofluorescence.
Reticulin antibodies have been classified by immunofluorescence into five types reacting with distinct antigens of intra- and extracellular components in mesenchyme. Two types of fibrillar antigens can be distinguished on the basis of the staining patterns, anatomical distribution, and species specificity. A third antibody reacts with either small fibres, amorphous proteins, or mucopolysaccharides lining the hepatic sinusoids (ground substance antigens). In addition, at least two kinds of intrasinusoidal cells show cytoplasmic fluorescence, ie, Kupffer cells and glass-adherent, blood-borne cells antigenically related to peritoneal macrophages. Some sera may contain antibodies reacting with sinusoidal endothelial cells though this has not yet been proven. It has been confirmed that all these distinct antibodies related to reticulin antigens are most frequent in dermatitis herpetiformis and coeliac disease, but they are also found with increased frequency in chronic heroin addicts and in rheumatoid and Sjögren's syndromes. About 5% of normal individuals had such antibodies and no significant increase could be demonstrated in autoimmune disorders or in liver cirrhosis. The antibodies appear to be stimulated by bacterial or nutritional antigens and are likely to represent anamnestic responses rather than direct cross reactions with a multiplicity of foreign antigens.


Adenovirus-mediated expression of Fas ligand induces hepatic apoptosis after Systemic administration and apoptosis of ex vivo-infected pancreatic islet allografts and isografts.
Fas ligand (FasL) mediates apoptosis of Fas-bearing cells and is expressed on a limited number of tissues, predominantly activated T lymphocytes. We describe the construction and biological activity of a replication-deficient type-5 adenovirus encoding murine FasL under the control of the cytomegalovirus (CMV) promoter (adCMV-FasL). In vitro, Jurkat cells undergo apoptosis when co-incubated with adCMV-FasL-infected COS cells. Systemic administration of adCMV-FasL to Wistar rats or DBA/2J mice results in widespread hepatic apoptosis and death in a dose-dependent manner within 72 hr, an effect not seen in lpr mice, or animals administered equivalent doses of adCMV-beta gal. Murine pancreatic islets also undergo apoptosis when infected ex vivo with adCMV-FasL, resulting in uniform primary nonfunction when transplanted into syngeneic or allogeneic diabetic recipients. These results indicate that adCMV-FasL is a potentially useful tool to study Fas/FasL biology.


Repetitive Mild Traumatic Brain Injury in Rats Impairs Cognition, Enhances Prefrontal Cortex Neuronal Activity, and Reduces Pre-synaptic Mitochondrial Function.
A major hurdle preventing effective interventions for patients with mild traumatic brain injury (mTBI) is the lack of known mechanisms for the long-term cognitive impairment that follows mTBI. The closed head impact model of repeated engineered rotational acceleration (rCHIMERA), a non-surgical animal model of repeated mTBI (rmTBI), mimics key features of rmTBI in humans. Using the rCHIMERA in rats, this study was designed to characterize rmTBI-induced behavioral disruption, underlying electrophysiological changes in the medial prefrontal cortex (mPFC), and associated mitochondrial dysfunction. Rats received 6 closed-head impacts over 2 days at 2 Joules of energy. Behavioral testing included automated analysis of behavior in open field and home-cage environments, rotarod test for motor skills, novel object recognition, and fear conditioning. Following rmTBI, rats spent less time grooming and less time in the center of the open field arena. Rats in their home cage had reduced inactivity time 1 week after mTBI and increased exploration time 1 month after injury. Impaired associative fear learning and memory in fear conditioning test, and reduced short-term memory in novel object recognition test were found 4 weeks after rmTBI. Single-unit in vivo recordings showed increased neuronal activity in the mPFC after rmTBI, partially attributable to neuronal disinhibition from reduced inhibitory synaptic transmission, possibly secondary to impaired mitochondrial function. These findings help validate this rat rmTBI model as replicating clinical features, and point to impaired mitochondrial functions after injury as causing imbalanced synaptic transmission and consequent impaired long-term cognitive dysfunction.


Use of polymer casts or metal particle infusion of ducts to study antigen uptake in the guinea pig mammary gland.
Microcorrosion casts were made of the duct system of guinea pig mammary glands by intramammary infusion of Mercox polyester resin following involution of the glands after the first lactation. The acinar configuration of the involuted gland was apparent on examination of the casts by scanning electron microscopy (SEM). Surface features, which were readily identified as those of imprints of ductal epithelium, were visible at higher magnifications. The morphology of these casts corresponded to the patterns observed by SEM of ethanol cryofractured specimens of mammary tissue. Cryofractured specimens of guinea pig mammary glands were also examined by SEM following intramammary infusion of tantalum. Tantalum particles were observed within the lumina of many ducts. Large phagocytic cells within the lumina were shown to contain tantalum by using back scatter imaging in conjunction with secondary imaging.


Functionally redundant formate dehydrogenases enable formate-dependent growth in Methanococcus maripaludis.
Methanogens are essential for the complete remineralization of organic matter in anoxic environments. Most cultured methanogens are hydrogenotrophic, using H2 as an electron donor to reduce CO2 to CH4, but in the absence of H2 many can also use formate. Formate dehydrogenase (Fdh) is essential for formate oxidation, where it transfers electrons for reduction of coenzyme F420 or to a flavin-based electron bifurcating reaction catalyzed by heterodisulfide reductase (Hdr), the terminal reaction of methanogenesis. Furthermore, methanogens that use formate encode at least two isoforms of Fdh in their genomes, but how these different isoforms participate in methanogenesis is unknown. Using Methanococcus maripaludis, we undertook a biochemical characterization of both Fdh isoforms involved in methanogenesis. Both Fdh1 and Fdh2 interacted with Hdr to catalyze the flavin-based electron bifurcating reaction, and both reduced F420 at similar rates. F420 reduction preceded flavin-based electron bifurcation activity for both enzymes. In a Δfdh1 mutant background, a suppressor mutation was required for Fdh2 activity. Genome sequencing revealed that this mutation resulted in loss of a specific molybdopterin transferase (moeA), allowing for Fdh2-dependent growth, and metal content of the proteins suggested that isoforms are dependent on either molybdenum or tungsten for activity. These data suggest that both isoforms of Fdh are functionally redundant, but their activities in vivo may be limited by gene regulation or metal availability under different growth conditions. Together these results expand our understanding of formate oxidation and the role of Fdh in methanogenesis.


Analysis of SecA dimerization in solution.
The Sec pathway mediates translocation of protein across the inner membrane of bacteria. SecA is a motor protein that drives translocation of preprotein through the SecYEG channel. SecA reversibly dimerizes under physiological conditions, but different dimer interfaces have been observed in SecA crystal structures. Here, we have used biophysical approaches to address the nature of the SecA dimer that exists in solution. We have taken advantage of the extreme salt sensitivity of SecA dimerization to compare the rates of hydrogen-deuterium exchange of the monomer and dimer and have analyzed the effects of single-alanine substitutions on dimerization affinity. Our results support the antiparallel dimer arrangement observed in one of the crystal structures of Bacillus subtilis SecA. Additional residues lying within the preprotein binding domain and the C-terminus are also protected from exchange upon dimerization, indicating linkage to a conformational transition of the preprotein binding domain from an open to a closed state. In agreement with this interpretation, normal mode analysis demonstrates that the SecA dimer interface influences the global dynamics of SecA such that dimerization stabilizes the closed conformation.


A randomized controlled trial testing an HIV prevention intervention for Latino youth.
To test the efficacy of a prevention intervention to reduce sexual risk behavior among Latino adolescents. Randomized controlled trial from April 2000 through March 2003, with data collection before and after intervention and at 3, 6, and 12 months. Northeast Philadelphia schools. Latinos aged 13 through 18 years (249 males and 304 females); 81.6% retained at 12-month follow-up. The HIV and health-promotion control interventions consisted of six 50-minute modules delivered by adult facilitators to small, mixed-gender groups in English or Spanish. Main Outcome Measure Self-reported sexual behavior. Analyses using generalized estimation equations over the follow-up period revealed that adolescents in the HIV intervention were less likely to report sexual intercourse (odds ratio, 0.66; 95% confidence interval [CI], 0.46-0.96), multiple partners (odds ratio, 0.53; 95% CI, 0.31-0.90), and days of unprotected intercourse (relative risk, 0.47; 95% CI, 0.26-0.84) and more likely to report using condoms consistently (odds ratio, 1.91; 95% CI, 1.24-2.93). Baseline sexual experience and language use moderated intervention efficacy. Adolescents assigned to the HIV intervention who were sexually inexperienced at baseline reported fewer days of unprotected sex (relative risk, 0.22; 95% CI, 0.08-0.63); Spanish speakers were more likely to have used a condom at last intercourse (odds ratio, 4.73; 95% CI, 1.72-12.97) and had a greater proportion of protected sex (mean difference, 0.35; P<.01) compared with similar adolescents in the health-promotion intervention. Results provide evidence for the efficacy of HIV intervention in decreasing sexual activity and increasing condom use among Latino adolescents.


POST-WAR GEOLOGY.
Believing as I do that geology in the immediate future will have a greater application, that with that application we shall see its expansion, I present the suggestions in this paper. They are based upon careful consideration, analyses, deductions from assembled views and opinions. Specialization will mean more and better geologists but also a greater graduate school population which must be fed from the undergraduate curriculum. There, and there particularly, is the basement complex on which to build. The greatest immediate need in geology is a broad, adequate undergraduate preparation, not so much geology itself, but in a slightly appalling list of collateral "must" subjects. Once this foundation is attained, graduate work in its several fields will care for itself.


The impact of phone counseling on urinary stone prevention.
We compared the effect of standard office-based consultation (OC) and phone correspondences (PC) on dietary 24-h urinary parameters. The medical record of all patients treated between January and April 2019 was reviewed. Only patients who had at least two consecutive 24-h urine collections were included. Linear and logistic regressions were used to investigate the difference between the changes in urinary parameters after OC and PC. Forty-three patients underwent 135 OC and 34 PC. Twenty-one received OC and PC, and 22 had only OC. Gender, age, the distance to stone clinic, the number of previous stone episodes, and baseline urinary parameters were similar between the groups. Patients who had both OC and PC had a longer follow-up time (51.7 vs 18.5 months, p < 0.0001) as well as more consults (Median 5.4 vs 2.5, p < 0.0001). Six (27%) patients who had only OC, and eight (38%) patients who had both OC and PC, experienced stone recurrence during the study period (p = 0.52). Following PC, there was a greater improvement in urine volume in comparison to OC (0.27 l/day vs -0.06 l/day, p = 0.034), but there was no difference in the absolute values after the consults between the groups. In established stone-clinic patients, PC was associated with a better adherence with follow-up. The 24-h urine results were similar between PC and OC. PC may be an effective alternative for urinary stone management.


Self-assembly of size-controlled liposomes on DNA nanotemplates.
Artificial lipid-bilayer membranes are valuable tools for the study of membrane structure and dynamics. For applications such as the study of vesicular transport and drug delivery, there is a pressing need for artificial vesicles with controlled size. However, controlling vesicle size and shape with nanometre precision is challenging, and approaches to achieve this can be heavily affected by lipid composition. Here, we present a bio-inspired templating method to generate highly monodispersed sub-100-nm unilamellar vesicles, where liposome self-assembly was nucleated and confined inside rigid DNA nanotemplates. Using this method, we produce homogeneous liposomes with four distinct predefined sizes. We also show that the method can be used with a variety of lipid compositions and probe the mechanism of templated liposome formation by capturing key intermediates during membrane self-assembly. The DNA nanotemplating strategy represents a conceptually novel way to guide lipid bilayer formation and could be generalized to engineer complex membrane/protein structures with nanoscale precision.


Fungicidal Impact of Salvadora Persica L. (Miswak) Extract on Growth of Foodborne Pathogens, Aspergillus Species.
Several investigations have focused on studying the suppressing influence of Salvadora persica (miswak) on oral microbes; however, studies regarding its fungicidal activity versus human aspergillosis-related illness are still scarce. The current research was designed to evaluate the fungicidal action of S persica aquatic root extract in terms of radial growth rate and inhibition zone (IZO) versus 3 pathogenic Aspergillus species, namely, A niger, A flavus, and A fumigatus in vitro. The results revealed that the plant extract (50 and 100 mg/mL) exhibited a prohibiting influence on the growth of the tested fungal species. The high concentration (100 mg/mL) of the plant extract was efficient in prohibiting the growing rate of the tested Aspergillus species after 6 days exposure period. Aspergillus niger and A flavus showed the largest inhibition ratios (60% and 54.4%, respectively) and IZO (33.00 ± 0.05 mm and 25.50 ± 0.18 mm, respectively) versus the control counterparts. Aspergillus fumigatus showed the minimum inhibition ratio (39%) and IZO (20.31 ± 0.05). The present data showed that the extract of S persica possesses potential fungicidal influence versus the tested pathogenic Aspergillus species and this may support the utilization of this extract as a promising antifungal agent versus aspergillosis.


Slow-channel depolarization: mechanism and control of arrhythmias.
The secondary inward current that flows through the slow channel is probably carried primarily by calcium ions. This current is responsible, in part, for the plateau phase of the cardiac action potential. Inward calcium current through the slow channel is essential to excitation-contraction coupling, and enhancement of this current exerts a positive inotropic effect. Transmembrane electrical potentials generated as a consequence of this slow inward current, so-called slow-channel depolarizations or slow responses, have been useful models in recent studies of cardiac autonomic interactions. The significance of the slow response in ventricular arrhythmias remains controversial and awaits more definitive experiments. The role of slow-channel depolarization as a basis for SA and AV nodal electrical activity is suggested by the electrophysiological similarities between these tissues and slow responses produced in vitro. This hypothesis is supported by the responses of these nodal tissues to interventions that augment of impede the slow inward current. More direct evidence in support of this notion may not be obtainable in the near future, since the critical voltage-clamp studies of ionic currents in SA and AV nodes are not technically feasible at this time.


The Effect of 12-Week Treatment with Intermittent Negative Pressure on Blood Flow Velocity and Flowmotion, Measured with a Novel Doppler Device (Earlybird). Secondary Outcomes from a Randomized Sham-Controlled Trial in Patients with Peripheral Arterial Disease.
Treatment with intermittent negative pressure (INP) is proposed as an adjunct to standard care in patients with peripheral arterial disease (PAD). The aims of this study were to evaluate the applicability of a novel ultrasound Dopplerdevice (earlybird) to assess blood flow characteristics in patients with PAD during a treatment session with INP, and whether certain flowproperties could determine who could benefit from INP treatment. Secondary outcomes of data from a randomized sham-controlled trial were explored. Patients were randomized to 12 weeks of treatment with 40 mm Hg or 10 mm Hg INP, for one hour twice daily. Earlybird blood flow velocity recordings were made before and after the 12-week treatmentperiod and consists of a 5-min recording in rest, 3-min during INP treatment and 5-min recording after ended INP test-treatment. Mean blood flow velocity (vmean), relative changes in flow and frequency spectrum by Fourier-transform of the respective bandwidths of endothelial, sympathetic, and myogenic functions, were analyzed for the different series of blood flow measurements. In total, 62 patients were eligible for analysis, where 32 patients were treated with 40 mm Hg INP. The acquired recordings were of good quality and were used for descriptive analyses of flow characteristics. An immediate increase in vmean during the negative pressure periods of the INP test-treatment was observed in the 40 mm Hg INP treatment group at both pre- and post-test. There was a significant difference between the treatment groups, with a difference between the medians of 13.7 (P < 0.001) at pre-test and 10.7 (P < 0.001) at posttest. This finding was confirmed with spectrum analysis by Fourier-transform of the bandwidth corresponding to INP treatment. The change in amplitude corresponding to myogenic function after 12-weeks of treatment, was significantly different in favor of the 40 mm Hg INP treatment group. We were not able to detect the specific flow characteristics indicating who would benefit INP-treatment. Earlybird is an applicable tool for assessing blood flow velocity in patients with PAD. Analysis of the flow velocity recordings shows that INP induces an immediate increase in blood flow velocities during INP. The positive effects of INP may be attributed to recruitment of arterioles, and thereby, increasing blood flow. In these analyses, no flow characteristics were determined which could predict who would benefit INP treatment.


Aldosterone blocks adrenal compensatory hypertrophy in the rat.
We report that adrenal compensatory hypertrophy occurs in intact and hypophysectomized anesthetized rats as well as in rats in which endogenous ACTH is suppressed by administration of dexamethasone or of dexamethasone plus low-dose ACTH. However, adrenal compensatory hypertrophy is blocked in intact and hypophysectomized animals when aldosterone alone or the combination of aldosterone, dexamethasone, and ACTH is administered using Alzet pumps. These data support previous reports that questioned the validity of the hypothesis that adrenal compensatory hypertrophy is controlled by the glucocorticoid-ACTH negative feedback system. These results require modification of current hypotheses concerning the mechanism of adrenal compensatory hypertrophy to allow for a central nervous system or other effect of aldosterone.


[Splenic artery embolization for the treatment of hypersplenism in hemophilic, HIV-1 and HCV seropositive patients].
Thrombocytopenia is an important and common hematological abnormality in patients with HIV-1/HCV coinfection. Splenomegaly is a frequent finding in these patients and usually causes hypersplenism and thrombocytopenia. We analyzed the clinical results of a minimal invasive treatment (splenic artery embolization) for thrombocytopenia secondary to hypersplenism and refractory to other therapies in two hemophiliac patients, HIV seropositive and with cirrhosis due to chronic HCV infection. The results suggest that splenic artery embolization is a safe, relatively atraumatic and effective method for the treatment of splenomegaly and hypersplenism in selected patients with HIV-1/HCV coinfection.


Experiments in digital literacy.
Having digital literacy requires more than just the ability to use software or to operate a digital device; it includes a large variety of complex skills such as cognitive, motoric, sociological, and emotional that users need to have in order to use digital environments effectively. A conceptual model that was recently described by the authors suggests that digital literacy comprises five major digital skills: photo-visual skills ("reading" instructions from graphical displays), reproduction skills (utilizing digital reproduction to create new, meaningful materials from preexisting ones), branching skills (constructing knowledge from non-linear, hypertextual navigation), information skills (evaluating the quality and validity of information), and socio-emotional skills (understanding the "rules" that prevail in cyberspace and applying this understanding in online cyberspace communication). The present paper presents results from a performance-based pioneer study that investigated the application of the above digital literacy skills conceptual model among different groups of scholars. Results clearly indicate that the younger participants performed better than the older ones, with photo-visual and branching literacy tasks, whereas the older participants were found to be more literate in reproduction and information literacy tasks. Research results shed light on the cognitive skills that users utilize in performing with digital environments, and provide educators and software developers with helpful guidelines for designing better user-centered digital environments.


Integrated optical device for Structured Illumination Microscopy.
Structured Illumination Microscopy (SIM) is a key technology for high resolution and super-resolution imaging of biological cells and molecules. The spread of portable and easy-to-align SIM systems requires the development of novel methods to generate a light pattern and to shift it across the field of view of the microscope. Here we show a miniaturized chip that incorporates optical waveguides, splitters, and phase shifters, to generate a 2D structured illumination pattern suitable for SIM microscopy. The chip creates three point-sources, coherent and controlled in phase, without the need for further alignment. Placed in the pupil of a microscope's objective, the three sources generate a hexagonal illumination pattern on the sample, which is spatially translated thanks to thermal phase shifters. We validate and use the chip, upgrading a commercial inverted fluorescence microscope to a SIM setup and we image biological sample slides, extending the resolution of the microscope.


Vascular endothelial growth factor induces expression of connective tissue growth factor via KDR, Flt1, and phosphatidylinositol 3-kinase-akt-dependent pathways in retinal vascular cells.
Fibroblastic proliferation accompanies many angiogenesis-related retinal and systemic diseases. Since connective tissue growth factor (CTGF) is a potent mitogen for fibrosis, extracellular matrix production, and angiogenesis, we have studied the effects and mechanism by which vascular endothelial growth factor (VEGF) regulates CTGF gene expression in retinal capillary cells. In our study, VEGF increased CTGF mRNA levels in a time- and concentration-dependent manner in bovine retinal endothelial cells and pericytes, without the need of new protein synthesis and without altering mRNA stability. VEGF activated the tyrosine receptor phosphorylation of KDR and Flt1 and increased the binding of phosphatidylinositol 3-kinase (PI3-kinase) p85 subunit to KDR and Flt1, both of which could mediate CTGF gene induction. VEGF-induced CTGF expression was mediated primarily by PI3-kinase activation, whereas PKC and ERK pathways made only minimal contributions. Furthermore, overexpression of constitutive active Akt was sufficient to induce CTGF gene expression, and inhibition of Akt activation by overexpressing dominant negative mutant of Akt abolished the VEGF-induced CTGF expression. These data suggest that VEGF can increase CTGF gene expression in bovine retinal capillary cells via KDR or Flt receptors and the activation of PI3-kinase-Akt pathway independently of PKC or Ras-ERK pathway, possibly inducing the fibrosis observed in retinal neovascular diseases.


A theoretical study of complexes between fullerenes and concave receptors with interest in photovoltaics.
The complexation of the pristine fullerenes C60 and C70 and the endohedral fullerenes Sc3N@C80 and Sc3N@C68 has been tested using a series of hosts of different nature, including the buckybowls corannulene and sumanene, a zinc porphyrin, a chloro boron subphthalocyanine, and a corannulene pentasubstituted with nitrile groups. A systematic theoretical study has been carried out in order to explore both the strength of the interaction and the feasibility for electron transfer of the dimers. Dispersion is the main stabilizing contribution in these dimers, so both molecules orientate so as to maximize the number of close contacts among atoms. As a consequence, all host molecules interact with C70 by the long axis. C60 and Sc3N@C80 are more spherically shaped, so there is no clear preference for the position of the host molecule, though endohedral fullerenes are encapsulated preferentially by the face without contacts with the inner cluster. Complexation energies increase with the contact surface between molecules in the complex. The most stable complexes with fullerenes are formed by the subphthalocyanine and the CN-pentasubstituted corannulene. Depending on the dimer, complexation energies span from around -15 kcal mol-1 of C60 with corannulene to -24 kcal mol-1 of Sc3N@C80 with the subphthalocyanine. Some of the dimers seem to be capable of acting as a donor-acceptor pair, leading to charge transfer states with a neat separation of charge, thus being candidates for organic photovoltaic devices. Endohedral fullerenes are less prone to these donor-acceptor transitions, with charge transfer taking place from the carbon cage to the endohedral cluster. cora5CN, with its inverted polarity, also shows charge transfer upon excitation but with the fullerene acting as a donor and the buckybowl as an acceptor.


Measuring spatial focusing in a migration system.
Equality indexes used in other geographical contexts may be used to gauge the degree of spatial focusing in an entire migration system or within the gross in- and out-migration fields of specific regions. They provide useful indicators of overall shifts in the patterns of interregional migration and can help give insight into the population redistributive roles played by specific regions. Perhaps the most common equality index used to measure income distribution is the Gini coefficient, yet it appears almost never to have been applied in migration research. In this paper we set forth a variety of Gini indexes to be used for different migration analyses and illustrate their application with recent data on U.S. interstate movements. We argue that the Gini index provides some singularly useful insights that differ from those afforded by other measures more commonly found to date in the migration analyst's tool kit.


Permanent diabetes during the first year of life: multiple gene screening in 54 patients.
The aim of this study was to investigate the genetic aetiology of permanent diabetes mellitus with onset in the first 12 months of age. We studied 46 probands with permanent, insulin-requiring diabetes with onset within the first 6 months of life (permanent neonatal diabetes mellitus [PNDM]/monogenic diabetes of infancy [MDI]) (group 1) and eight participants with diabetes diagnosed between 7 and 12 months of age (group 2). KCNJ11, INS and ABCC8 genes were sequentially sequenced in all patients. For those who were negative in the initial screening, we examined ERN1, CHGA, CHGB and NKX6-1 genes and, in selected probands, CACNA1C, GCK, FOXP3, NEUROG3 and CDK4. The incidence rate for PNDM/MDI was calculated using a database of Italian patients collected from 1995 to 2009. In group 1 we found mutations in KCNJ11, INS and ABCC8 genes in 23 (50%), 9 (19.5%) and 4 (8.6%) patients respectively, and a single homozygous mutation in GCK (2.1%). In group 2, we identified one incidence of a KCNJ11 mutation. No genetic defects were detected in other loci. The incidence rate of PNDM/MDI in Italy is estimated to be 1:210,287. Genetic mutations were identified in ~75% of non-consanguineous probands with PNDM/MDI, using sequential screening of KCNJ11, INS and ABCC8 genes in infants diagnosed within the first 6 months of age. This percentage decreased to 12% in those with diabetes diagnosed between 7 and 12 months. Patients belonging to the latter group may either carry mutations in genes different from those commonly found in PNDM/MDI or have developed an early-onset form of autoimmune diabetes.


Reliability of the perceptual evaluation of adductor spasmodic dysphonia.
Although perceptual assessment by experienced voice clinicians remains the gold standard for the diagnosis and assessment of severity of adductor spasmodic dysphonia (ADSD), the interrater reliability of voice experts for this task has not been assessed. In addition, it is unknown whether telephone-recorded or -transmitted voice samples could be used for this task. The aims of this study were (1) to assess the reliability of perceptual analysis of ADSD severity by voice experts and (2) to compare the results between digitally recorded voice samples and those recorded over the telephone. Five laryngologists randomly selected voice samples from 46 ADSD patients and rated the severity of ADSD on a 5-point rating scale. A set of digital voice recordings and a set of telephone voice recordings made from filtering the digital set via the telephone were rated, and each voice set was rated twice. Measures of intrarater and interrater reliability, as well as a measure of the probability of agreement among the raters, were calculated. There was a high level of agreement on ADSD severity, with excellent interrater and intrarater reliability (Cronbach's alpha, .93 to .96). The probabilities of rater agreement on the digitally recorded and telephone-filtered voice samples were similar (chi2, p = .07). The ratings of digital versus telephone voice samples were highly correlated (Pearson r, 0.99; p < .001). These results demonstrate that voice experts are reliably able to judge and agree on the severity of ADSD. Telephone-filtered voices appear to convey adequate ADSD perceptual cues for expert listeners to judge the severity of spasmodic dysphonia.


Insect-inspired neuromorphic computing.
The steady improvement in the performance of computing systems seen for many decades is levelling off as the miniaturization of semiconducting technology approaches the atomic limit, facing severe physical and technological issues. Neuromorphic computing is an emerging solution which makes use of silicon technology in a different way, inline with the computational principles observed in animal nervous systems. In this article, we argue that the nervous systems of insects in particular offer themselves as an ideal starting point for incorporation into realistic neuromorphic systems and review research in developing insect-inspired neuromorphic systems. We conclude with an exciting yet tangible vision of a full neuromorphic sensory-motor system where a liquid state machine modelling the function of the insect mushroom body links input to output and allows for amalgamation of the work discussed in a hierarchical framework of a full system inspired by the concept of how information flows through insects.


Interleukin-32: a new proinflammatory cytokine involved in hepatitis C virus-related liver inflammation and fibrosis.
Interleukin 32 (IL-32) is a recently described proinflammatory cytokine that activates p38 mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB), thereby inducing proinflammatory cytokines such as IL-1β and tumor necrosis factor alpha (TNF-α). We investigated the role of IL-32 in patients with chronic hepatitis C virus (HCV) infection. Steady-state hepatic messenger RNA (mRNA) levels of IL-32 were determined in a cohort of 90 subjects; anti-IL-32 staining was used in a second cohort of 132 consecutive untreated chronic HCV patients. Correlations with histological features of steatosis, inflammation, and fibrosis were made. In vitro, endogenous IL-32 in monocytes and in the human hepatoma cell line Huh-7.5 were examined. The effects of IL-32-overexpression and IL-32-silencing on HCV replication were studied using HCV luciferase reporter viruses. There were highly significant positive associations between hepatic IL-32 mRNA expression and liver steatosis, inflammation, fibrosis, smooth muscle actin (SMA) area, and serum alanine aminotransferase (ALT) levels. IL-32 protein expression was positively associated with portal inflammation, SMA area, and ALT. In vitro, IL-1β and TNF-α significantly induced IL-32 expression in human Huh-7.5 cells. Alone, stimulation with interferon alpha (IFN-α) did not induce IL-32 expression in Huh-7.5. However, IFN-α exerted a significant additive effect on TNF-α-induced but not IL-1β-induced IL-32 expression, particularly in CD14+ monocytes. This effect was dependent both on NF-κB and Jak/STAT signaling. Viral infection of Huh-7.5 cells resulted in a significant (11-fold) induction of IL-32 mRNA expression. However, modulation of IL-32 in Huh-7.5 cells by overexpression or silencing did not influence HCV virus replication as determined by luciferase assays. IL-32 is a novel proinflammatory cytokine involved in HCV-associated liver inflammation/fibrosis. IL-32 is expressed by human hepatocytes and hepatoma cells and its expression is regulated by proinflammatory stimuli.


[The toxicological characteristics of the interaction of cholinolytics with aminostigmine--a new reversible cholinesterase inhibitor].
This investigation has been carried out on albino mice. In the first series of experiments, we determined the dependence of LD50 of aminostigmine on different doses of 10 cholinolytics, and conversely, the dependence of LD50 of cholinolytics on aminostigmine administration. The initial slopes of the dose-effect curves were calculated. This data form the basis for evaluation of the character and degree of interactions. We established that benactyzine, spasmolytin, ftoracizin, arpenal, atropine, ganglerone, and methacin at low doses exhibit antagonism (with decreasing activity), whereas at high doses they exhibit synergism to the toxic effect of aminostigmine. In the interaction with aminostigmine, pirenzepine and amitriptyline (M1--cholinolytics) reveal a slightly pronounced antagonism, whereas aetyrophene, a selective central N-cholinolytic, displays mutual synergism. In the second series of experiments we showed that a combination of M1-, M2-cholinolytic atropine with M1- cholinolytics does not change the efficacy of the prophylaxis of aminostigmine and physostigmine poisoning, whereas a combination with N-cholinolytic increases it.


Ocular Surface Disease in Glaucoma: Effect of Polypharmacy and Preservatives.
To evaluate the prevalence of ocular surface disease (OSD) in glaucoma and nonglaucoma subjects using different clinical tests and to determine the effect of number of antiglaucoma medications and preservatives on OSD. This is a cross-sectional, case-comparison study at the Eye Clinic of the University of Malaya Medical Centre, Malaysia, between June 2012 and January 2013. Glaucoma subjects (n = 105) using topical antiglaucoma medications were compared with control subjects (n = 102) who were not on any topical medications. The presence of OSD was assessed using the tear film breakup time (TBUT) test, corneal staining, Schirmer test, and Ocular Surface Disease Index (OSDI) questionnaire grading. The prevalence of OSD varied from 37 to 91% in the glaucoma group, depending on the type of clinical test. More subjects in the glaucoma group had corneal staining (63% vs. 36%, p = 0.004), abnormal Schirmer tests (39% vs. 25%, p = 0.049), and moderate OSDI symptoms (17% vs. 7%, p = 0.028). The percentage with abnormal TBUT increased with higher numbers of topical medications and was high with both benzalkonium chloride-containing and preservative-free eye drops (90% and 94%, respectively, both p < 0.001). Benzalkonium chloride was associated with a nearly three times higher odds ratio of showing abnormal OSDI. Ocular surface disease is common in those using topical antiglaucoma medications. Abnormal TBUT is associated with increasing number of eye drops and benzalkonium chloride-containing eye drops, although this also occurs with the use of preservative-free eye drops.


Longitudinal speech perception performance of young children with cochlear implants and tactile aids plus hearing aids.
Nineteen profoundly deaf children who utilize either cochlear implants (CIs) or multichannel tactile aids plus hearing aids (T + HAs) and who were enrolled in a full-day educational program that specializes in the use of sensory aids were evaluated using a battery of speech perception tests either developed in house or chosen because they were part of a standard battery of tests developed for children with sensory aids. The tests were organized into four perceptual levels ranging from pattern perception at level one to open set word identification at level four. For each level, data were analyzed for changes over time and for differences between performance of CI children and those using T + HAs. The results indicate that overall, across levels, both groups improved significantly over time but no differences were found between users of T + HAs and CIs at any level.


[Prolonged or chronic fatigue of unknown origin].
Although prolonged or chronic fatigue is a very common complaint in primary care medicine, a biomedical obvious cause is often not found. In such a case, for women between 18 and 50 years with a ferritin level of less than 50 µg/l in the absence of anaemia, an iron supplementation may be associated with an improvement in fatigue. Appropriate treatment is also important for depression, anxiety or insomnia. In other cases, the approach is essentially non-pharmacological in the form of lifestyle advice, empathy and cognitive behavioural therapy as well as progressive and adapted physical exercises.


The Building Blocks Collaborative: advancing a life course approach to health equity through multi-sector collaboration.
Too many children are born into poverty, often living in disinvested communities without adequate opportunities to be healthy and thrive. Two complementary frameworks-health equity and life course-propose new approaches to these challenges. Health equity strategies seek to improve community conditions that influence health. The life course perspective focuses on key developmental periods that can shift a person's trajectory over the life course, and highlights the importance of ensuring that children have supports in place that set them up for long-term success and health. Applying these frameworks, the Alameda County Public Health Department launched the Building Blocks Collaborative (BBC), a countywide multi-sector initiative to engage community partners in improving neighborhood conditions in low-income communities, with a focus on young children. A broad cross-section of stakeholders, called to action by the state of racial and economic inequities in children's health, came together to launch the BBC and develop a Bill of Rights that highlights the diverse factors that contribute to children's health. BBC partners then began working together to improve community conditions by learning and sharing ideas and strategies, and incubating new collaborative projects. Supportive health department leadership; dedicated staff; shared vision and ownership; a flexible partnership structure; and broad collective goals that build on partners' strengths and priorities have been critical to the growth of the BBC. Next steps include institutionalizing BBC projects into existing infrastructure, ongoing partner engagement, and continued project innovation-to achieve a common vision that all babies have the best start in life.


Proton leak and hydrogen peroxide production in liver mitochondria from energy-restricted rats.
Energy restriction (ER), without malnutrition, is the only environmental intervention that consistently increases maximum life span in laboratory rodents. One theory proposes that a reduction in energy expenditure and reactive oxygen species production is the mechanism responsible for this action of ER. To further test this theory, proton leak, H2O2 production, lipid peroxidation, and protein carbonyls were measured in mitochondria from FBNF1 rats fed either a control or 40% ER diet (onset at 6 mo of age). Liver mitochondria were isolated at 7 and 12 mo of age. Liver weight decreased 25 and 36% at 1 and 6 mo of ER, respectively (P < 0.05). ER resulted in an increase (P < 0.05) in percent total polyunsaturates, n-6 polyunsaturates, and total unsaturates (6 mo only) in mitochondrial lipids. These changes, however, were not associated with significant alterations in mitochondrial function. State 4 respiration and membrane potential were not different (P > 0.05) between groups at either assessment period. Similarly, proton leak kinetics were not different between control and ER animals. Top-down metabolic control analysis and its extension, elasticity analysis, were used at the 6-mo assessment and revealed no difference in control of the oxidative phosphorylation system between control and ER rats. H2O2 production with either succinate or pyruvate/malate substrates was also not different (P > 0.05) between groups at either time point. In conclusion, ER did not alter proton leak or H2O2 production at this age or stage of restriction in liver.


Glutamine flux imaging using genetically encoded sensors.
Genetically encoded sensors allow real-time monitoring of biological molecules at a subcellular resolution. A tremendous variety of such sensors for biological molecules became available in the past 15 years, some of which became indispensable tools that are used routinely in many laboratories. One of the exciting applications of genetically encoded sensors is the use of these sensors in investigating cellular transport processes. Properties of transporters such as kinetics and substrate specificities can be investigated at a cellular level, providing possibilities for cell-type specific analyses of transport activities. In this article, we will demonstrate how transporter dynamics can be observed using genetically encoded glutamine sensor as an example. Experimental design, technical details of the experimental settings, and considerations for post-experimental analyses will be discussed.


Age-related changes in lymphocyte subsets of quarter horse foals.
To characterize changes in lymphocyte subsets over time in foals from birth to 18 weeks of age, accounting for differences among individuals, and to determine the effect of overnight storage of blood samples on foal lymphocyte subset concentrations. 8 healthy Quarter Horse foals from birth to 18 weeks of age. Blood samples were collected longitudinally from birth to 18 weeks of age and a CBC performed on each sample. The samples were stained for lymphocyte markers, either immediately or after overnight storage and analyzed by flow cytometry. Total leukocytes, total lymphocytes, and the absolute concentrations of all lymphocyte subsets increased significantly with age. The proportions of B29A+, CD21+, and-equine major histocompatability complex class-II molecule+ lymphocytes increased significantly with age. The proportion of equine (Eq) CD5+, EqCD8+, and EqWC4+ lymphocytes decreased significantly with age. Significant differences among foals were found with respect to initial concentrations with respect to initial concentrations, but not with respect to the rate of increase of the various subsets tested. Significant differences were not found in subset values when comparing blood samples stained on the day of collection or after overnight storage at room temperature (approx 21 C) or under refrigeration. These results are consistent with an increase in subset numbers and proportions over time, but with individual differences among foals. The observation of individual differences in subsets among foals suggests that there may be individual differences in susceptibility to infectious disease during the perinatal period. The absence of an effect of overnight storage makes field studies of lymphocyte subset concentrations more feasible.


Satellite cell proliferation and myofiber cross-section area increase after electrical stimulation following sciatic nerve crush injury in rats.
Electrical stimulation has been recommended as an effective therapy to prevent muscle atrophy after nerve injury. However, the effect of electrical stimulation on the proliferation of satellite cells in denervated muscles has not yet been fully elucidated. This study was aimed to evaluate the changes in satellite cell proliferation after electrical stimulation in nerve injury and to determine whether these changes are related to the restoration of myofiber cross-section area (CSA). Sciatic nerve crush injury was performed in 48 male Sprague-Dawley rats. In half (24/48) of the rats, the gastrocnemius was electrically stimulated transcutaneously on a daily basis after injury, while the other half were not stimulated. Another group of 24 male Sprague-Dawley rats were used as sham operation controls without injury or stimulation. The rats were euthanized 2, 4, and 6 weeks later. After 5-bromo-2'-deoxyuridine (BrdU) labeling, the gastrocnemia were harvested for the detection of paired box protein 7 (Pax7), BrdU, myofiber CSA, and myonuclei number per fiber. All data were analyzed using two-way analysis of variance and Bonferroni post-hoc test. The percentages of Pax7-positive nuclei (10.81 ± 0.56%) and BrdU-positive nuclei (34.29 ± 3.87%) in stimulated muscles were significantly higher compared to those in non-stimulated muscles (2.58 ± 0.33% and 1.30 ± 0.09%, respectively, Bonferroni t = 15.91 and 18.14, P < 0.05). The numbers of myonuclei per fiber (2.19 ± 0.24) and myofiber CSA (1906.86 ± 116.51 μm) were also increased in the stimulated muscles (Bonferroni t = 3.57 and 2.73, P < 0.05), and both were positively correlated with the Pax7-positive satellite cell content (R = 0.52 and 0.60, P < 0.01). There was no significant difference in the ratio of myofiber CSA/myonuclei number per fiber among the three groups. Our results indicate that satellite cell proliferation is promoted by electrical stimulation after nerve injury, which may be correlated with an increase in myonuclei number and myofiber CSA.


A prospective study on drug monitoring of PEGasparaginase and Erwinia asparaginase and asparaginase antibodies in pediatric acute lymphoblastic leukemia.
This study prospectively analyzed the efficacy of very prolonged courses of pegylated Escherichia coli asparaginase (PEGasparaginase) and Erwinia asparaginase in pediatric acute lymphoblastic leukemia (ALL) patients. Patients received 15 PEGasparaginase infusions (2500 IU/m(2) every 2 weeks) in intensification after receiving native E coli asparaginase in induction. In case of allergy to or silent inactivation of PEGasparaginase, Erwinia asparaginase (20 000 IU/m(2) 2-3 times weekly) was given. Eighty-nine patients were enrolled in the PEGasparaginase study. Twenty (22%) of the PEGasparaginase-treated patients developed an allergy; 7 (8%) showed silent inactivation. The PEGasparaginase level was 0 in all allergic patients (grade 1-4). Patients without hypersensitivity to PEGasparaginase had serum mean trough levels of 899 U/L. Fifty-nine patients were included in the Erwinia asparaginase study; 2 (3%) developed an allergy and none silent inactivation. Ninety-six percent had at least 1 trough level ≥100 U/L. The serum asparagine level was not always completely depleted with Erwinia asparaginase in contrast to PEGasparaginase. The presence of asparaginase antibodies was related to allergies and silent inactivation, but with low specificity (64%). Use of native E coli asparaginase in induction leads to high hypersensitivity rates to PEGasparaginase in intensification. Therefore, PEGasparaginase should be used upfront in induction, and we suggest that the dose could be lowered. Switching to Erwinia asparaginase leads to effective asparaginase levels in most patients. Therapeutic drug monitoring has been added to our ALL-11 protocol to individualize asparaginase therapy.


[Delirium due to intoxication with quetiapine: a systematic literature review].
Quetiapine is a frequently prescribed antipsychotic and therefore often used in overdose. Delirium (with anticholinergic delirium as a specific condition) is described as a serious complication of quetiapine intoxication.
 AIM: To assess the scientific literature on delirium as a side effect of quetiapine intoxication: incidence, symptoms and treatment.
 METHOD: A systematic Medline literature search.
 RESULTS: The systematic literature search resulted in 36 papers: 11 cohort studies, 24 case reports (22 papers) and 3 review papers. The reported incidence varied greatly, probably due to different quality of assessment. The clinical picture is characterized by a varying combination of peripheral and central symptoms, with agitation occurring frequently. Treatment is mainly supportive. Physostigmine is described as a specific treatment for anticholinergic delirium/toxidrome. Effectiveness of other pharmacological interventions remains unclear.
 CONCLUSION: Delirium due to quetiapine intoxication is described repeatedly. Presumably there is an underreporting of this condition and associated symptoms. Better knowledge could lead to better detection and treatment.


Identification of the neighborhood and CA rules from spatio-temporal CA patterns.
Extracting the rules from spatio-temporal patterns generated by the evolution of cellular automata (CA) usually produces a CA rule table without providing a clear understanding of the structure of the neighborhood or the CA rule. In this paper, a new identification method based on using a modified orthogonal least squares or CA-OLS algorithm to detect the neighborhood structure and the underlying polynomial form of the CA rules is proposed. The Quine-McCluskey method is then applied to extract minimum Boolean expressions from the polynomials. Spatio-temporal patterns produced by the evolution of 1D, 2D, and higher dimensional binary CAs are used to illustrate the new algorithm, and simulation results show that the CA-OLS algorithm can quickly select both the correct neighborhood structure and the corresponding rule.


Silver-stained nucleolar organizer regions in the differentiation of prostatic hyperplasia, intraepithelial neoplasia, and adenocarcinoma.
Nucleolar organizer regions (NORs) are loops of chromosomal DNA that ultimately direct the development of the nucleolus. These NORs are associated with argyrophilic acidic nonhistone proteins which have allowed the demonstration of NORs by a simple silver stain. This staining technique has been used to examine the number of silver-stained NORs (AgNORs) in malignant and benign conditions, and, in general, AgNOR numbers are increased in malignant tissues when compared with normal or benign conditions. More recently, this technique has been applied to premalignant lesions with varying results. We applied this silver stain to prostate lesions where nucleolar features are of diagnostic importance. The stain was applied to ten cases of prostatic hyperplasia, ten cases each of prostatic intraepithelial neoplasia (PIN) grades 1 through 3, and ten cases each of low grade, intermediate grade, and high grade adenocarcinoma. The counting was performed by image analysis. A significant mean difference did exist between low grade and the two higher grades of adenocarcinoma. There was no significant difference in AgNOR counts per nucleus in hyperplasia, PIN, or adenocarcinoma or between the three grades of PIN. The AgNOR silver stain is not useful in the differentiation of hyperplasia, the three grades of PIN, and adenocarcinoma. Also because of overlap among the three grades of adenocarcinoma, it is not useful in the differentiation of these lesions.


Phosphatidylinositol 4,5-Bisphosphate Modulates the Affinity of Talin-1 for Phospholipid Bilayers and Activates Its Autoinhibited Form.
Integrins are vital transmembrane receptors that mediate cell-cell and cell-extracellular matrix interactions and signaling. Talin is a 270 kDa protein and is considered a key regulator of integrin activity. The interaction between talin and integrin is commonly regarded as the final step of inside-out activation. In the cytosol, talin adopts an autoinhibited conformation, in which the C-terminal rod domain binds the N-terminal head domain, preventing the interactions of the head domain with the membrane surface and the integrin cytoplasmic domain. It has long been suggested that the presence of phosphatidylinositol 4,5-bisphosphate (PIP2) at focal adhesions plays a role in activating talin. However, a detailed picture and mechanism of PIP2 activation of autoinhibited talin remains elusive. Here, we use a fluorescence resonance energy transfer-based binding assay to measure the affinity of talin and lipid bilayers harboring anionic lipids. Results show that the R9 and R12R13 segments of the talin rod domain inhibit the binding of the talin head domain (THD) to anionic lipid bilayers. In contrast, we show that the binding of the THD to bilayers containing PIP2 is insensitive to the presence of the inhibitor domains, thereby directly implicating PIP2 as an effective activator of talin. Furthermore, we have mapped the activation to the interaction of PIP2 with the F2F3 domain of the talin head, showing that PIP2 plays a critical role in the regulation of the autoinhibited form of talin and stimulates recruitment of talin to the membrane, which is essential for integrin inside-out signaling.


Fundamentals of Endoscopic Surgery cognitive examination: development and validity evidence.
Flexible endoscopy is an integral part of surgical care. Exposure to endoscopic procedures varies greatly in surgical training. The Society of American Gastrointestinal and Endoscopic Surgeons has developed the Fundamentals of Endoscopic Surgery (FES), which serves to teach and assess the fundamental knowledge and skills required to practice flexible endoscopy of the gastrointestinal tract. This report describes the validity evidence in the development of the FES cognitive examination. Core areas in the practice of gastrointestinal endoscopy were identified through facilitated expert focus groups to establish validity evidence for the test content. Test items then were developed based on the content areas. Prospective enrollment of participants at various levels of training and experience was used for beta testing. Two FES cognitive test versions then were developed based on beta testing data. The Angoff and contrasting group methods were used to determine the passing score. Validity evidence was established through correlation of experience level with examination score. A total of 220 test items were developed in accordance with the defined test blueprint and formulated into two versions of 120 questions each. The versions were administered randomly to 363 participants. The correlation between test scores and training level was high (r = 0.69), with similar results noted for contrasting groups based on endoscopic rotation and endoscopic procedural experience. Items then were selected for two test forms of 75 items each, and a passing score was established. The FES cognitive examination is the first test with validity evidence to assess the basic knowledge needed to perform flexible endoscopy. Combined with the hands-on skills examination, this assessment tool is a key component for FES certification.


Interaction between Social/Psychosocial Factors and Genetic Variants on Body Mass Index: A Gene-Environment Interaction Analysis in a Longitudinal Setting.
Obesity, which develops over time, is one of the leading causes of chronic diseases such as cardiovascular disease. However, hundreds of BMI (body mass index)-associated genetic loci identified through large-scale genome-wide association studies (GWAS) only explain about 2.7% of BMI variation. Most common human traits are believed to be influenced by both genetic and environmental factors. Past studies suggest a variety of environmental features that are associated with obesity, including socioeconomic status and psychosocial factors. This study combines both gene/regions and environmental factors to explore whether social/psychosocial factors (childhood and adult socioeconomic status, social support, anger, chronic burden, stressful life events, and depressive symptoms) modify the effect of sets of genetic variants on BMI in European American and African American participants in the Health and Retirement Study (HRS). In order to incorporate longitudinal phenotype data collected in the HRS and investigate entire sets of single nucleotide polymorphisms (SNPs) within gene/region simultaneously, we applied a novel set-based test for gene-environment interaction in longitudinal studies (LGEWIS). Childhood socioeconomic status (parental education) was found to modify the genetic effect in the gene/region around SNP rs9540493 on BMI in European Americans in the HRS. The most significant SNP (rs9540488) by childhood socioeconomic status interaction within the rs9540493 gene/region was suggestively replicated in the Multi-Ethnic Study of Atherosclerosis (MESA) (p = 0.07).


Food for the Future.
There is ample justification for concern about adequate food supplies for future generations of mankind, but this concern should be broadened to include the immediate problem of an adequate standard of living for the world's present population. The successful solution of the immediate problems would furnish the best background of experience for meeting those that will arise in the future. First steps include the acceptance by society of responsibilities for the extension of the benefits of education throughout the world and provision for the type of scientific, economic, social, political, and religious leadership necessary to assure food for all on a continuing basis. Striking improvements in the food supply can be readily made through the application of present knowledge, if the foregoing conditions are met. The rapid pace of modern science, both pure and applied, gives promise that future benefits may be much greater than those thus far experienced. Current advances do not signal the end of a technical road but rather that the great scientific developments still lie ahead. If we have the intelligence and wisdom to recognize human responsibilities and to make constructive use of our natural and human resources, we can look forward to a better world in the future and improved standards of living for all.