Coronary Artery Disease in Patients with Critical Limb Ischemia Undergoing Major Amputation or Not.
Due to the increase of elderly and diabetes patients, surgeons encounter patients requiring treatment of critical limb ischemia (CLI) in the presence of systemic arteriosclerotic diseases. In this study, we retrospectively investigated the prevalence of coronary artery disease (CAD) in patients with CLI who underwent major (above-the-ankle) amputation or nonmajor amputation (below-the-ankle amputation or debridement of wound). We retrospectively investigated 129 consecutive patients surgically managed for CLI in our institution between January 2013 and December 2015. The prevalence of CAD was defined as a cardiac treatment history or significant vascular stenosis (stenosis of > 75%). The outcomes were compared between patients who underwent major amputation (n = 36) and nonmajor amputation (n = 93). Additionally, archived record of 566 patients treated nonsurgically by percutaneous transluminal angioplasty in our institution was investigated to evaluate patients with milder peripheral artery disease. CAD was present in 83 patients (69%), including 82% of patients who underwent major amputation and 63% of nonmajor amputation group. The prevalence of CAD was significantly higher in the major amputation group (P = 0.042). Ejection fraction was not significantly different (P > 0.05). Among the 566 CLI patients treated by only percutaneous transluminal angioplasty, 227 (40%) had CAD, which was a significantly lower prevalence than those surgically treated (P < 0.001). The presence of CAD is more frequent in CLI patients who require extended surgical management of the limb than in those who do not. Evaluation of CAD and careful perioperative management are important for patients with CLI patients.


Unusually large sialolith of Wharton's duct.
The formation of calcific concretions in the salivary duct or glands is a common disorder, especially in the submandibular glands. Most of the salivary calculi are small in size, in contrast to those that reach several centimeters, which are reported as megaliths or giant calculi in the literature. They may occur in any of the salivary gland ducts but are most common in Wharton's duct and the submandibular gland. This report presents clinical and radiographical sign of an unusually large sialolith. There was painless swelling on the floor of the edentulous mouth and patient was unaware of it. Radiographical examination revealed large irregular radio-opaque mass superimposed on right canine and premolar areas. This case report describes a patient presenting with an unusually large submandibular gland duct sialolith, the subsequent patient management, the aetiology, diagnosis and its treatment.


Effect of mechanical loading on periodontal cells.
Mechanical loading is an important regulatory factor in alveolar bone homeostasis, and plays an essential role in maintaining the structure and mass of the alveolar processes throughout lifetime. A better understanding of the cellular and molecular responses of periodontal cells is a prerequisite for further improvements of therapeutic approaches in orthodontics, periodontal and alveolar bone repair and regeneration, implantology, and post-surgical wound healing. The purpose of this review is to provide an insight into some cell culture and animal models used for studying the effects of mechanical loading on periodontal cells, and into the recent developments and utilization of new in vivo animal models. There has been an increased awareness about the need for improvement and development of in vivo models to supplement the widely used cell culture models, and for biological validation of in vitro results, especially in the light of evidence that developmental models may not always reflect bone homeostasis in an adult organism. Due to the limitations of in vivo models, previous studies on mechanical regulation of alveolar bone osteoblasts and cementoblasts mostly focused on proliferative responses, rather than on the stimulation of cell differentiation. To address this problem, we have recently characterized and implemented a mouse osteoinductive tooth movement model for studying mechanically induced regulation of osteoblast- and cementoblast-associated genes. In this model, a defined and reproducible mechanical osteogenic loading is applied during a time course of up to two weeks. Regulation of gene expression in either wild-type or transgenic animals is assessed by a relative quantitative measurement of the level of target mRNAs directly within the subpopulations of periodontal cells. To date, results demonstrate a defined temporal pattern of cell-specific gene regulation in periodontal osteoblasts mechanically stimulated to differentiate and deposit bone matrix. The responses of osteoblast-associated genes to mechanical loading were 10- to 20-fold greater than the increase in the numbers of these cells, indicating that the induction of differentiation and an increase of cell function are the primary responses to osteogenic loading. The progression of the osteoblast phenotype in the intact mouse periodontium was several-fold faster compared with that in cultured cells, suggesting that the mechanical signal may be targeting osteoblast precursors in the state of readiness to respond to an environmental challenge, without the initial proliferative response. An early response of alkaline phosphatase and bone sialoprotein genes was detected after 24 hrs of treatment, followed by a concomitant stimulation of osteocalcin and collagen I between 24 and 48 hrs, and deposition of osteoid after 72 hrs. Although cementoblasts constitutively express biochemical markers similar to those of osteoblasts, distinct responses of osteocalcin, collagen I, and bone sialoprotein genes to mechanical loading were observed in the two cell phenotypes. This finding indicates that differential genetic responses to mechanical loading provide functional markers for distinction of the cementoblast and osteoblast phenotypes.


Executive functions are employed to process episodic and relational memories in children with autism spectrum disorders.
Long-term memory functioning in autism spectrum disorders (ASDs) is marked by a characteristic pattern of impairments and strengths. Individuals with ASD show impairment in memory tasks that require the processing of relational and contextual information, but spared performance on tasks requiring more item-based, acontextual processing. Two experiments investigated the cognitive mechanisms underlying this memory profile. A sample of 14 children with a diagnosis of high-functioning ASD (age: M = 12.2 years), and a matched control group of 14 typically developing (TD) children (age: M = 12.1 years), participated in a range of behavioral memory tasks in which we measured both relational and item-based memory abilities. They also completed a battery of executive function measures. The ASD group showed specific deficits in relational memory, but spared or superior performance in item-based memory, across all tasks. Importantly, for ASD children, executive ability was significantly correlated with relational memory but not with item-based memory. No such relationship was present in the control group. This suggests that children with ASD atypically employed effortful, executive strategies to retrieve relational (but not item-specific) information, whereas TD children appeared to use more automatic processes. The relational memory impairment in ASD may result from a specific impairment in automatic associative retrieval processes with an increased reliance on effortful and strategic retrieval processes. Our findings allow specific neural predictions to be made regarding the interactive functioning of the hippocampus, prefrontal cortex, and posterior parietal cortex in ASD as a neural network supporting relational memory processing.


8-Hydroxydaidzein Induces Apoptosis and Inhibits AML-Associated Gene Expression in U-937 Cells: Potential Phytochemical for AML Treatment.
8-hydroxydaidzein (8-OHD) is a compound derived from daidzein, known for its anti-inflammatory and anti-proliferative properties in K562 human chronic myeloid leukemia (CML) cells. However, its effects on acute myeloid leukemia (AML) cells have not been fully understood. To investigate its potential anti-AML mechanism, we employed an integrated in vitro-in silico approach. Our findings demonstrate that 8-OHD suppresses the expression of CDK6 and CCND2 proteins and induces cell apoptosis in U-937 cells by activating Caspase-7 and cleaving PARP-1. Microarray analysis revealed that 8-OHD downregulates differentially expressed genes (DEGs) associated with rRNA processing and ribosome biogenesis pathways. Moreover, AML-target genes, including CCND2, MYC, NPM1, FLT3, and TERT, were downregulated by 8-OHD. Additionally, molecular docking software predicted that 8-OHD has the potential to interact with CDK6, FLT3, and TERT proteins, thereby reducing their activity and inhibiting cell proliferation. Notably, we discovered a synergic pharmacological interaction between 8-OHD and cytarabine (Ara-C). Overall, this study provides insights into the therapeutic applications of 8-OHD in treating AML and elucidates its underlying mechanisms of action.


Continuous synthesis of carbon dots with full spectrum fluorescence and the mechanism of their multiple color emission.
Carbon dots with different emission fluorescence have a great number of potential applications for various areas from in vitro imaging and biotherapy, due to the good biosafety of red fluorescent CDs, to efficient ion detection and photocatalysis, due to the excellent photoluminescence properties of blue fluorescent carbon dots. Traditional methods for the synthesis of full-spectrum carbon dots require 24 h of synthesis and complex column chromatography. In this paper, a facile and efficient microfluidic method to continuously synthesize small and uniform carbon dots with full-spectrum emission fluorescence is developed for the first time. The synthesis process could be reduced to 20 minutes. Through XPS analysis and DFT calculations, it is quantitatively revealed that the number of primary amino groups determines the energy gap of the carbon dots and thus determines the fluorescence emission wavelength of the carbon dots. Applications for precise Fe3+ detection and in vitro bio-imaging were successfully implemented, showing great potential application value of the carbon dots.


Patterns and controls of the dynamics of net primary production by understory macroalgal assemblages in giant kelp forests.
Macroalgae are important primary producers in many subtidal habitats, yet little information exists on the temporal and spatial dynamics of net primary production (NPP) by entire subtidal assemblages. This knowledge gap reflects the logistical challenges in measuring NPP of diverse macroalgal assemblages in shallow marine habitats. Here, we couple a simple primary production model with nondestructive estimates of taxon-specific biomass on subtidal reefs off Santa Barbara, California to produce a 4-year time series of net primary production by intact assemblages of understory macroalgae in giant kelp forests off Santa Barbara, California, USA. Daily bottom irradiance varied significantly throughout the year, and algal assemblages were on average exposed to saturating irradiance for only 1.3-4.5 h per day, depending on the time of year. Despite these variable light-limiting conditions, biomass rather than irradiance explained the vast majority of variation observed in daily NPP at all times of the year. Measurements of peak biomass in spring and summer proved to be good predictors of NPP for the entire year, explaining as much as 76% of the observed variation. In contrast, bottom irradiance was a poor predictor of NPP, explaining <10% of the variation in NPP when analyzed seasonally and ~2% when evaluated annually. Our finding that annual NPP by macroalgal assemblages can be predicted from a single, nondestructive measurement of biomass should prove useful for developing time series data that are necessary to evaluate natural and anthropogenic changes in NPP by one of the world's most productive ecosystems.


Breast cancer.
Several issues of clinical importance were in the forefront of discussion during 1989. Heated debates continue regarding whether alcohol, oral contraceptives or hormonal replacement therapy modifies a patient's risk for developing breast cancer. It is quite apparent, and rightfully so, that a major impetus in the arena of research in breast cancer seems to be towards cure of the disease. The rationale for offering adjuvant systemic treatment to a larger group of primary breast cancer patients who have a potential for cure certainly seems justifiable. However, one still needs to individualize patients based on their prognosis and objectively weigh the potential benefits and risks of adjuvant therapy in a given clinical situation, especially in patients with less than 1 cm primary tumors and negative axillary nodes as they have been shown to have a 20-year recurrence-free survival of 86%. A better understanding of several molecular and genetic parameters has begun. Further developments in this area may lead to clinically appropriate means of preventing or treating this disease. Metastatic disease continues to remain an incurable entity. Efforts aimed at treating metastatic disease with more 'intensive dose chemotherapy' have not been demonstrated to be beneficial for affected patients.


[Use of angiotensin II receptor blockaders in heart failure].
Angiotensin converting enzyme (ACE) inhibitors and diuretics represent the first line of therapy in patients with symptomatic heart failure. Inhibition of angiotensin II production is, however, incomplete with ACE inhibitors, due to non-ACE dependent conversion pathways. Moreover, some patients are intolerant to ACE inhibitors due to side effects or renal insufficiency. Angiotensin II receptor blockers may be an alternative to, or an additional treatment in heart failure. Preliminary studies comparing the angiotensin II receptor blocker losartan with placebo have demonstrated improved haemodynamic parameters, reduced hospitalisation and mortality in patients with heart failure. Reduced morbidity and mortality have also been found with losartan treatment, as compared to the ACE inhibitor captopril. This paper discusses the role of angiotensin II receptor blockers in the treatment of heart failure. Some results from published studies and a short description of ongoing trials are presented.


IGF-1 Influences Gonadotropin-Releasing Hormone Regulation of Puberty.
The pubertal process is initiated as a result of complex neuroendocrine interactions within the preoptic and hypothalamic regions of the brain. These interactions ultimately result in a timely increase in the secretion of gonadotropin-releasing hormone (GnRH). Researchers for years have believed that this increase is due to a diminished inhibitory tone which has applied a prepubertal brake on GnRH secretion, as well as to the gradual development of excitatory inputs driving the increased release of the peptide. Over the years, insulin-like growth factor-1 (IGF-1) has emerged as a prime candidate for playing an important role in the onset of puberty. This review will first present initial research demonstrating that IGF-1 increases in circulation as puberty approaches, is able to induce the release of prepubertal GnRH, and can advance the timing of puberty. More recent findings depict an early action of IGF-1 to activate a pathway that releases the inhibitory brake on prepubertal GnRH secretion provided by dynorphin, as well as demonstrating that IGF-1 can also act later in the process to regulate the synthesis and release of kisspeptin, a potent stimulator of GnRH at puberty.


Mapping local matrix remodeling induced by a migrating tumor cell using three-dimensional multiple-particle tracking.
Mesenchymal cell migration through a three-dimensional (3D) matrix typically involves major matrix remodeling. The direction of matrix deformation occurs locally in all three dimensions, which cannot be measured by current techniques. To probe the local, 3D, real-time deformation of a collagen matrix during tumor cell migration, we developed an assay whereby matrix-embedded beads are tracked simultaneously in all three directions with high resolution. To establish a proof of principle, we investigated patterns of collagen I matrix deformation near fibrosarcoma cells in the absence and presence of inhibitors of matrix metalloproteinases and acto-myosin contractility. Our results indicate that migrating cells show patterns of local matrix deformation toward the cell that are symmetric in magnitude with respect to the axis of cell movement. In contrast, patterns of matrix release from the cell are asymmetric: the matrix is typically relaxed first at the back of the cell, allowing forward motion, and then at the cell's leading edge. Matrix deformation in regions of the matrix near the cell's leading edge is elastic and mostly reversible, but induces irreversible matrix rupture events near the trailing edge. Our results also indicate that matrix remodeling spatially correlates with protrusive activity. This correlation is mediated by myosin II and Rac1, and eliminated after inhibition of pericellular proteolysis or ROCK. We have developed an assay based on high-resolution 3D multiple-particle tracking that allows us to probe local matrix remodeling during mesenchymal cell migration through a 3D matrix and simultaneously monitor protrusion dynamics.


Clinical and Dermoscopic Features Associated With Difficult-to-Recognize Variants of Cutaneous Melanoma: A Systematic Review.
The clinical and dermoscopic features of the vast majority of uncommon variants of cutaneous melanoma have been rarely reported, leading to difficulty in making accurate diagnoses. To define the main clinical and dermoscopic features of the most frequently reported but uncommon histologic variants of cutaneous melanoma. A 2-step systematic review of the literature was performed (from inception to November 2018) using PubMed, Embase, and Cochrane Central Register of Controlled Trials databases. The first step identified those uncommon variants of melanoma for which at least 1 case reporting dermoscopy was described in the literature. The keywords searched were melanoma, uncommon, rare, dermoscopy, and dermatoscopy. In the second step, each previously identified uncommon variant was searched for in the same databases by combining the following terms with melanoma, dermoscopy, and dermatoscopy: amelanotic, hypopigmented, animal, melanocytoma, balloon, desmoplastic, follicular, nested, nevoid, dermal, spitz*, spindle, and verrucous. The institution's database was also searched from January 2012 to September 2019 for histopathologically confirmed cases of the same melanoma variants. Each reviewer also assessed the quality of reporting in the included articles based on previously described guidelines. In total, 62 articles met the inclusion criteria, reporting 433 melanoma cases. An additional 56 cases of uncommon melanoma variants were retrieved from the institution's database for a total of 489 cases: 283 cases of amelanotic superficial spreading melanoma, 18 cases of animal-type and pigmented epithelioid melanocytoma, 7 cases of balloon cell melanoma, 71 cases of desmoplastic melanoma, 3 cases of follicular melanoma, 10 cases of nested melanoma, 33 cases of nevoid melanoma, 2 cases of primary dermal melanoma, 57 cases of spitzoid melanoma, and 5 cases of verrucous melanoma. These variants of melanoma occurred more frequently in women than men (147 cases vs 132 cases). Clinically, these tumors were mainly palpable (162 of 217 [74.7%]) or amelanotic (283 of 489 [57.9%]) lesions that could resemble other benign or malignant skin conditions; dermoscopy typically revealed a homogeneous pinkish background, white structures, and polymorphic vessels. The mean age of all included was 58 years (range, 1-89 years). Uncommon melanoma variants may resemble both inflammatory disorders and other cutaneous neoplasms, representing a diagnostic pitfall even for the most experienced dermatologist. The purpose of this systematic review was to provide an extensive and detailed overview of specific clinical and dermoscopic features of each uncommon melanoma variant, highlighting the main criteria for differentiating these variants from other benign or malignant skin lesions.


Review of tracheostomy videos for staff education.
Tracheostomy care is a basic nursing skill. While it is a matter of routine procedure in the daily practice of otolaryngology and critical care nurses, general nurses in other areas may perform it infrequently. Therefore, many otolaryngology nurses provide staff education when a tracheostomy patient is present in a nonspecialty area. This educational effort can be time consuming and repetitious for the specialty nurse. Audiovisual materials can be useful as a basis for an educational session or as a substitute when the specialty nurse is unavailable. The author conducted a thorough search for available videorecordings on nursing care of adults with tracheostomies. Four videos were identified and acquired. Each one was viewed twice, and then subjectively rated for completeness and accuracy of content in a comparative, "Consumer Reports"-like manner. Recommendations are made considering content, cost, and availability. The review is written for experienced clinicians and presumes a thorough knowledge of tracheostomy care.


Coupled binding and folding of intrinsically disordered proteins: what can we learn from kinetics?
Protein or protein regions that are not forming well-defined structures in their free states under native-like conditions are called intrinsically disordered proteins. Such proteins are very common in protein-protein interactions, where their disorder apparently gives several advantages including optimal binding properties. To fully appreciate why protein disorder is advantageous for protein-protein interactions we need to understand the mechanism(s) of interaction. However, elucidating mechanisms in protein-protein interactions is usually very challenging. Here we discuss how kinetics in combination with protein engineering and structural information can be used to depict details of protein-protein interactions involving intrinsically disordered proteins.


Social cognition in schizophrenia: cognitive and affective factors.
Social cognition refers to how people conceive, perceive, and draw inferences about mental and emotional states of others in the social world. Previous studies suggest that the concept of social cognition involves several abilities, including those related to affect and cognition. The present study analyses the deficits of individuals with schizophrenia in two areas of social cognition: Theory of Mind (ToM) and emotion recognition and processing. Examining the impairment of these abilities in patients with schizophrenia has the potential to elucidate the neurophysiological regions involved in social cognition and may also have the potential to aid rehabilitation. Two experiments were conducted. Both included the same five tasks: first- and second-level false-belief ToM tasks, emotion inferencing, understanding of irony, and matrix reasoning (a WAIS-R subtest). The matrix reasoning task was administered to evaluate and control for the association of the other tasks with analytic reasoning skills. Experiment 1 involved factor analysis of the task performance of 75 healthy participants. Experiment 2 compared 30 patients with schizophrenia to an equal number of matched controls. Results. (1) The five tasks were clearly divided into two factors corresponding to the two areas of social cognition, ToM and emotion recognition and processing. (2) Schizophrenics' performance was impaired on all tasks, particularly on those loading heavily on the analytic component (matrix reasoning and second-order ToM). (3) Matrix reasoning, second-level ToM (ToM2), and irony were found to distinguish patients from controls, even when all other tasks that revealed significant impairment in the patients' performance were taken into account. The two areas of social cognition examined are related to distinct factors. The mechanism for answering ToM questions (especially ToM2) depends on analytic reasoning capabilities, but the difficulties they present to individuals with schizophrenia are due to other components as well. The impairment in social cognition in schizophrenia stems from deficiencies in several mechanisms, including the ability to think analytically and to process emotion information and cues.


Determinants of overweight with concurrent stunting among Ghanaian children.
Malnutrition (undernutrition and overnutrition) is a major public health problem in Ghana -affecting growth and development of individuals and the nation. Stunting and overweight are of particular interest, as recent national surveys show a rising trend of overnutrition and stubbornly high burden of stunting among Ghanaian children. There are currently no data on the simultaneous occurrence of overweight and stunting within individuals in Ghana. This paper presents the burden, the individual-level, and contextual determinants of overweight with concurrent stunting among Ghanaian children. This study analyzed data set of the fourth round of the Ghana Multiple Indicator Cluster Survey (MICS4). Bivariate analyses were used to describe selected characteristics of survey respondents and their children. Hierarchical modelling approach facilitated identification of significant distal, intermediate and proximal factors/determinants of concurrent stunting and overweight. Both crude and adjusted prevalence ratios via a multivariable Poison regression model with their corresponding 95% Confidence Intervals (CI) are reported. Variables with p ≤ 0.25 at the bivariate level were included in the multivariable analysis. An alpha value of 5% was used to indicate significance. Of 7550 cases (children) analyzed, the prevalence of stunting was 27.5%; underweight was 17.3%; and wasting was 7.7%. The prevalence of overweight and concurrent overweight and stunting were respectively 2.4% and 1.2%. Children who belonged to the fourth wealth quintile, were more likely to be overweight and concurrently stunted as against children belonging to the poorest quintile (aPR = 1.010; 95% CI, 1.003-1.017). Compared to religious (Christians/Muslim/Traditionalist) household heads, children whose household heads did not belong to any religion had 2 times the rates of the Overweight with concurrent stunting (PR = 2.024; 95% CI, 1.016-4.034). Children with mothers aged 20-34 and 35-49 had an increased though insignificant prevalence ratio of association (aPR = 1.001; 95% CI, 0.994-1.005) and (aPR = 1.001; 95% CI, 0.998-1.012) respectively. This analysis determined the prevalence of concurrent stunting and overweight among Ghanaian children to be 1.2%. Four contextual variables (breastfeeding status, religion, geographic region, and wealth index quintile) were associated with overweight with concurrent stunting. We conclude that, only contextual factors are predictive of DBM among children under five living in Ghana.


Correlation of serum VEGF-C, ANGPTL4, and activin A levels with frailty.
Secretory factors linked to lymphogenesis, such as vascular endothelial growth factor C (VEGF-C), angiopoietin like protein 4 (ANGPTL4), and activin A (ACV-A), have been recognized as potential markers of chronic inflammatory status and age-related diseases. Furthermore, these factors may also be linked to frailty. The primary objective of this study was to examine the serum VEGF-C, ANGPTL4, and ACV-A levels in young individuals, healthy older individuals, and older individuals with pre-frailty and frailty, and to determine their association with pro-inflammatory factor levels. We conducted an observational study, enrolling a total of 210 older individuals and 20 young healthy volunteers. Participants were divided into four groups based on the Freid frailty phenotype: healthy young group, older patients without frailty group, pre-frail older group, and frail older group. Plasma and peripheral blood mononuclear cells (PBMCs) were collected from all four groups. ELISA was used to measure the serum levels of VEGF-C, ANGPTL4, ACV-A, and pro-inflammatory cytokines, while RT-qPCR was used to measure the transcription level of VEGF-C, ANGPTL4 and ACV-A in PBMCs. In comparison to healthy young individuals and older participants without frailty, older participants with frailty exhibited lower renal function, higher serum levels and transcription levels of VEGF-C, ANGPTL4, ACV-A, and elevated levels of pro-inflammatory cytokines (CRP, IL-1β, and TNF-α). Multiple linear regression analysis revealed that serum levels of VEGF-C, ANGPTL4, and ACV-A were positively correlated with the frailty index, independent of age, eGFR, and comorbidities. Furthermore, the receiver operating characteristic (ROC) curve analysis demonstrated that serum levels of VEGF-C, ANGPTL4, and ACV-A have great accuracy in predicting frailty. Elevated serum levels of VEGF-C, ANGPTL4, and ACV-A are associated with frailty status.


Transition of chemolithotroph Ferrobacillus ferrooxidans to obligate organotrophy and metabolic capabilities of glucose-grown cells.
Transition of chemolithotrophic Ferrobacillus ferrooxidans to organotrophy occurred after 60 hr of incubation in an organic medium. Three distinct phases, based on metabolic activities of cells, were observed during the course of transition. Conversion of cellular nutrition to organotrophy resulted in a gradual loss of Fe(2+) oxidation and cessation of CO(2) fixation. These changes were concomitant with a rapid increase in uptake of glucose and phosphate during the latter part of transition period. The outcome of transition was governed by the pH of the medium, temperature of incubation, availability of oxygen, age of the chemolithotrophic cells, and the type of energy and carbon source available to the bacterium. Presence or absence of p-aminobenzoic acid and Fe(2+) ions did not influence transition of cells. A defined medium containing glucose, mineral salts, and p-aminobenzoic acid at pH 2.5 was found to be most suitable for transition and for culture of heterotrophic convertants. Maximum growth rate of the heterotrophic cells was attained with vigorous aeration at 35 C. The bacterium could be cultured on a variety of organic compounds, including complex organic media, provided they were used in low concentrations. Serological studies on autotrophic cells and the heterotrophic convertant have shown a definite antigenic relationship between the two cell types.


Significant variation between SNP-based HLA imputations in diverse populations: the last mile is the hardest.
Four single nucleotide polymorphism (SNP)-based human leukocyte antigen (HLA) imputation methods (e-HLA, HIBAG, HLA*IMP:02 and MAGPrediction) were trained using 1000 Genomes SNP and HLA genotypes and assessed for their ability to accurately impute molecular HLA-A, -B, -C and -DRB1 genotypes in the Human Genome Diversity Project cell panel. Imputation concordance was high (>89%) across all methods for both HLA-A and HLA-C, but HLA-B and HLA-DRB1 proved generally difficult to impute. Overall, <27.8% of subjects were correctly imputed for all HLA loci by any method. Concordance across all loci was not enhanced via the application of confidence thresholds; reliance on confidence scores across methods only led to noticeable improvement (+3.2%) for HLA-DRB1. As the HLA complex is highly relevant to the study of human health and disease, a standardized assessment of SNP-based HLA imputation methods is crucial for advancing genomic research. Considerable room remains for the improvement of HLA-B and especially HLA-DRB1 imputation methods, and no imputation method is as accurate as molecular genotyping. The application of large, ancestrally diverse HLA and SNP reference data sets and multiple imputation methods has the potential to make SNP-based HLA imputation methods a tractable option for determining HLA genotypes.


Approaches for studying the pathogenic T cells in autoimmune patients.
Our provisional conclusions from this work are as follows. (1) For screening responses of established lines, native human AChR is not prohibitively scarce, especially if it is concentrated onto beads, and class II-transfected TE671 cells may be useful too; both may give vital evidence of AChR-specificity, but it is still crucial to confirm that with synthetic peptides. (2) For mapping epitopes, panels of full-length and shorter recombinant human polypeptides, and of synthetic peptides, are invaluable complementary material: longer peptides tend to stimulate particularly strongly. (3) Initial selection with pooled synthetic peptides can easily generate interesting lines from both patients and controls, but they may depend on the artificial processing sites that are an inevitable consequence of arbitrarily chosen start and stop points. Of course, these might conceivably be employed in unusual antigen-presenting cells (such as thymic myoid cells), so we cannot totally dismiss such "cryptic" epitopes. This system can sometimes select T cells responding to "natural" epitopes too, as now reported for tetanus toxin. Nevertheless, for these and other reasons, at present, we strongly favor using the longest human recombinant material possible, because it is apparently processed more naturally. This must be combined with rigorous screening for reactivity to E. coli-derived contaminants plus concomitant mapping of epitopes as above. Use of intact AChR for initiating lines may yet become feasible. (4) The T cells thus isolated and characterized so far are proving to be heterogeneous in the epitopes and presenting class II molecules they recognize, and in their T-cell receptor gene usage. It is premature to claim key myasthenogenic epitopes or clonotypes, but HLA-DR3 and the linked -DQw2 do not appear to monopolize presentation. (5) Assessing the disease-relevance of these T cells is a separate problem, highlighted by their apparent similarity in healthy controls. In the meantime, to test their potential pathogenicity, we are assaying their cytokine profiles and ability to help specific antibody production in vitro. In the hope that they do prove to be relevant, we are also using some of them to test possible therapeutic strategies that might prove applicable in the patients.


Towards a unified framework to study causality in Earth-life systems.
There is considerable interest in better understanding how earth processes shape the generation and distribution of life on Earth. This question, at its heart, is one of causation. In this article I propose that at a regional level, earth processes can be thought of as behaving somewhat deterministically and may have an organized effect on the diversification and distribution of species. However, the study of how landscape features shape biology is challenged by pseudocongruent or collinear variables. I demonstrate that causal structures can be used to depict the cause-effect relationships between earth processes and biological patterns using recent examples from the literature about speciation and species richness in montane settings. This application shows that causal diagrams can be used to better decipher the details of causal relationships by motivating new hypotheses. Additionally, the abstraction of this knowledge into structural equation metamodels can be used to formulate theory about relationships within Earth-life systems more broadly. Causal structures are a natural point of collaboration between biologists and Earth scientists, and their use can mitigate against the risk of misassigning causality within studies. My goal is that by applying causal theory through application of causal structures, we can build a systems-level understanding of what landscape features or earth processes most shape the distribution and diversification of species, what types of organisms are most affected, and why.


Racial Differences in Prevalence and Risk for Intracranial Atherosclerosis in a US Community-Based Population.
Intracranial atherosclerotic disease (ICAD) is an important cause of stroke; however, little is known about racial differences in ICAD prevalence and its risk factors. To determine racial differences in ICAD prevalence and the risk factors (both midlife and concurrent) associated with its development in a large, US community-based cohort. Analysis of 1752 black and white participants recruited from the Atherosclerosis Risk in Communities (ARIC) cohort study who underwent 3-dimensional intracranial vessel wall magnetic resonance imaging from October 18, 2011 to December 30, 2013; data analysis was performed from October 18, 2011 to May 13, 2015. Midlife and concurrent cardiovascular risk factors. Intracranial plaque presence, size (maximum normalized wall index) and number were assessed by vessel wall magnetic resonance imaging. Midlife and concurrent vascular risk factor associations were determined by Poisson regression (plaque presence), negative binominal regression (plaque number), and linear regression (plaque size), and compared between races. Of the 1752 study participants (mean [SD] age, 77.6 [5.3] years; range, 67-90 years), 1023 (58.4%) were women and 518 (29.6%) were black. Black men had the highest prevalence (50.9% vs 35.9% for black women, 35.5% for white men, and 30.2% for white women; P < .001) and the highest frequency (22.4% vs 12.1% for black women, 10.7% for white men, and 8.7% for white women; P < .01) of multiple plaques. Prevalence increased with age, reaching 50% before ages 68, 84, and 88 years in black men, white men, and white women, respectively (ICAD prevalence remained <50% in black women). Midlife hypertension and hyperlipidemia were associated with 29% (prevalence ratio [PR], 1.29; 95% CI, 1.08-1.55) and 18% (PR, 1.18; 95% CI, 0.98-1.42), respectively, increased ICAD prevalence with no significant differences between races. Midlife hypertension was also associated with larger plaques (1.11 higher mean maximum normalized wall index; 95% CI, 0.21-2.01). Midlife smoking and diabetes were associated with increased risk of ICAD in black individuals (102% [PR, 2.02; 95% CI, 1.12-3.63] and 57% [PR, 1.57; 95% CI, 1.13- 2.19], respectively), but not in white participants (P < .05 interaction by race). The prevalence of ICAD was highest in black men. Midlife smoking and diabetes were strongly associated with late-life ICAD in blacks only, whereas midlife hypertension and hyperlipidemia were associated with late-life ICAD in both races. These associations may help to explain racial differences in US stroke rates and offer insight into preventive risk-factor management strategies.


NMR relaxation characteristics of rubidium-87 in perfused rat salivary glands.
Rubidium is a good substitute for potassium in many biological systems, and it has been suggested that rubidium-87 nuclear magnetic resonance (87Rb-NMR) spectroscopy could be used to measure K+ fluxes across membranes in intact tissues. To evaluate this possibility, isolated rat mandibular salivary glands were perfused with solutions containing Rb+ in place of K+. The 87Rb signals arising from the intra- and extracellular compartments were first separated by spectral subtraction and then subjected to line-shape analysis. The narrow extracellular signal was a single Lorentzian (line-width 156 Hz), whereas the broader intracellular signal consisted of two Lorentzian components (ca. 530 and 3080 Hz). Double-quantum filtering of the 87Rb signal from the glands revealed two components of transverse relaxation in antiphase (rate constants 1.8 and 13.3 ms-1), showing the probable involvement of quadrupolar interactions in the relaxation of intracellular Rb+. We conclude, therefore, that both line-shape analysis and double-quantum filtering could provide a basis for the measurement of unidirectional K+ fluxes in intact tissues.


Endothelial activation by angiotensin II through NFkappaB and p38 pathways: Involvement of NFkappaB-inducible kinase (NIK), free oxygen radicals, and selective inhibition by aspirin.
Angiotensin-II (AII), the dominant effector of the renin-angiotensin system, is involved in the pathogenesis of cardiovascular diseases, such as atherosclerosis. Upregulation of the adhesion molecules VCAM-1, ICAM-1, and E-selectin in endothelial cells by inflammatory cytokines through nuclear factor kappa B (NFkappaB) activation is implicated in formation and progression of atherosclerotic plaque. Here we show that AII induces NFkappaB-dependent transcription in primary endothelial cell lines, leading to the upregulation of ICAM-1 and VCAM-1 expression. NFkappaB activation by AII is mediated by the NFkappaB-inducing kinase (NIK), a common mediator of NFkappaB activation by inflammatory cytokines, such as TNF-alpha. However, NFkappaB stimulation by AII differs from that of TNF-alpha since a TNF-receptor associated factor 2 (TRAF-2) dominant negative mutant does not prevent AII-mediated NFkappaB activation. In analogy with TNF-alpha-dependent activation of NFkappaB, treatment with either the anti-oxidant N-acetyl cysteine (NAC) or the cyclooxygenase (COX) inhibitor acetyl salicylic acid (aspirin), but not indometacin, prevents the induction of NFkappaB-dependent transcription by AII. Thus, production of reactive oxygen species, aspirin (asp)-sensitive enzymes of the arachidonate metabolism, and NIK are common transducers of AII- and TNF-dependent pathways to NFkappaB. AII also activates the inflammatory p38 kinase in endothelial cells, an effect inhibited by exposure to either NAC or asp. Pharmacological interference of the p38 pathway, with the inhibitor SB 202190, prevented AII-mediated activation of the NFkappaB target V-CAM, without affecting degradation of IkappaBalpha. These results support a pro-inflammatory effect of the vasoactive peptide AII in endothelial cells, through at least two pathways-NFkappaB and p38-both of which are sensitive to asp and antioxidants.


The use of cannabis and perceptions of its effect on fertility among infertility patients.
What is the prevalence of cannabis use and the perceptions of its impact on fertility among infertility patients? A total of 13% of infertility patients used cannabis within the last year, and current usage is associated with patient perceptions of negative effects of cannabis on fertility and pregnancy. Cannabis use is increasing among the general population and pregnant women, particularly in places where cannabis use is legal despite having known and potential negative effects on fertility and pregnancy. A cross-sectional patient survey study was performed between July 2017 and September 2017. Patients attending a university-affiliated hospital-based fertility clinic (n = 290) were invited to complete a written survey. Inclusion criteria were limited to the ability to read English. There were no exclusion criteria. Of the 290 patients approached, 270 (93%) agreed to participate. The questions covered demographics, cannabis usage, perceptions of the effect of cannabis on fertility and pregnancy, cessation of use due to infertility and personal history of disclosing cannabis use to healthcare providers (HCP). The results showed that 13% of respondents disclosed use of cannabis in the past year (past year users) and 38% had not used cannabis in the past year but had previously used cannabis (>1 year users) while 49% had never used cannabis (never users). Baseline demographics were similar for the three groups, but across four measures of fertility and pregnancy health, past-year users perceived less of a negative effect compared to >1 year users, and never users (P values of 0.02, 0.03, 0.01, <0.001 for questions on pregnancy, offspring health, male fertility and female fertility, respectively). Of past year users, 72% said they had or would disclose use to their HCP, but only 9.4% reported that their HCP had actually instructed them to discontinue use. Self-reported patient surveys are subject to reporting bias and may not reflect actual use and perceptions. This study suggests that cannabis use is common among infertility patients. Given the known negative impacts of cannabis on pregnancy, the authors would have expected informed infertility patients to cease cannabis use as part of their efforts to conceive. As the prevalence of cannabis use in the last year among infertility patients is similar to that in the general Canadian population, it is unclear whether the prevalence of cannabis use in the sample population merely reflects the average usage in society or, after taking into account those who reduced their usage to improve their fertility, is a factor contributing to infertility and thus prompting fertility referral. Given concern about the potential negative impact of cannabis use on fertility, and that only 9% of past year users had been instructed by an HCP to cease cannabis use, HCPs should consider the benefits of counselling about cannabis cessation for patients who are attempting to conceive. Future research should focus on analysing the effects of cannabis use on female fertility and determining whether a reduction in use among patients with infertility can improve conception rates. Michelle Shin, Clinical Research Associate, is supported by the University of Toronto GREI Fellowship Fund, which is sponsored by unrestricted research grants from EMD Serono, Merck Canada and Ferring Pharmaceuticals. The authors have no potential conflicts of interest to disclose.


Axonal caliber and neurofilaments are proportionately decreased in galactose neuropathy.
Feeding galactose to rats induces nerve conduction abnormalities, increased levels of nerve galactitol, endoneurial edema, elevated pressure and hypoxia of endoneurial fluid, and pathological abnormalities of nerve fibers. To investigate the cellular mechanisms of the fiber lesions and their possible relationship to alterations in the nerve microenvironment, rat peroneal nerves were morphometrically evaluated eight months after the commencement of galactose feeding. Whereas the density of neurofilaments (NF/micron2) in the transverse axonal area of myelinated fibers was not significantly different between the nerves of galactose-fed and control rats, axonal areas and the number of NF/axon, when related to myelin spiral length, were significantly less in nerves of galactose-fed rats. Myelin alterations, characteristic of axonal atrophy, were also significantly increased. The present data provide evidence of a proportionate decrease in axonal caliber and the number of NF/axon in myelinated fibers in experimental galactose neuropathy, suggesting that galactose induces fibers in experimental galactose neuropathy, suggesting that galactose induces either decreased NF synthesis, assembly or transport. The possible role of microenvironmental alterations, including endoneurial hypoxia and hyperosmolarity, in the production of this axonal atrophy is discussed.


Pharmacodynamic Effects of Topical Omiganan in Patients With Mild to Moderate Atopic Dermatitis in a Randomized, Placebo-Controlled, Phase II Trial.
Omiganan is an indolicidin analog with antimicrobial properties that could be beneficial for patients with atopic dermatitis. In this randomized, double-blind, placebo-controlled, phase II trial we explored the efficacy, pharmacodynamics, and safety of topical omiganan once daily in 36 patients with mild to moderate atomic dermatitis. Patients were randomized to apply topical omiganan 1%, omiganan 2.5%, or vehicle gel to one target lesion once daily for 28 consecutive days. Small but significant improvements in local objective SCORing Atopic Dematitis index and morning itch were observed in the omiganan 2.5% group compared with the vehicle gel group (-18.5%; 95% confidence interval, -32.9 to -1.0; P = 0.04; and -8.2; 95% confidence interval, -16.3 to -0.2; P = 0.05, respectively). A shift from lesional to nonlesional skin microbiota was observed in both omiganan treatment groups, in contrast to the vehicle group. Thus, treatment with topical omiganan improved dysbiosis in patients with mild to moderate atopic dermatitis, and small but statistically significant improvements in clinical scores were detected. Our findings warrant further exploration in future clinical trials.


Clustering of two genes putatively involved in cyanate detoxification evolved recently and independently in multiple fungal lineages.
Fungi that have the enzymes cyanase and carbonic anhydrase show a limited capacity to detoxify cyanate, a fungicide employed by both plants and humans. Here, we describe a novel two-gene cluster that comprises duplicated cyanase and carbonic anhydrase copies, which we name the CCA gene cluster, trace its evolution across Ascomycetes, and examine the evolutionary dynamics of its spread among lineages of the Fusarium oxysporum species complex (hereafter referred to as the FOSC), a cosmopolitan clade of purportedly clonal vascular wilt plant pathogens. Phylogenetic analysis of fungal cyanase and carbonic anhydrase genes reveals that the CCA gene cluster arose independently at least twice and is now present in three lineages, namely Cochliobolus lunatus, Oidiodendron maius, and the FOSC. Genome-wide surveys within the FOSC indicate that the CCA gene cluster varies in copy number across isolates, is always located on accessory chromosomes, and is absent in FOSC's closest relatives. Phylogenetic reconstruction of the CCA gene cluster in 163 FOSC strains from a wide variety of hosts suggests a recent history of rampant transfers between isolates. We hypothesize that the independent formation of the CCA gene cluster in different fungal lineages and its spread across FOSC strains may be associated with resistance to plant-produced cyanates or to use of cyanate fungicides in agriculture.


CD1d-restricted natural killer T cells can down-regulate tumor immunosurveillance independent of interleukin-4 receptor-signal transducer and activator of transcription 6 or transforming growth factor-beta.
It has been shown previously that the suppression of tumor immunosurveillance may be a mechanism by which tumors resist immune detection and elimination. In this study, we evaluated the role of the immunoregulatory natural killer T (NKT) cells in the biology of immunosurveillance of osteosarcoma. The K7M2 mouse osteosarcoma cell line was implanted orthotopically into wild-type and NKT cell-deficient CD1d knockout (KO) BALB/c mice, and mice were monitored for growth of primary tumors. Further, we examined the role of CD4(+) and/or CD8(+) cells by depleting the cells in vivo and measuring CTL activity in vitro. We also asked the role of interleukin (IL)-4 receptor alpha (IL-4Ralpha)-signal transducer and activator of transcription 6 (STAT6) signaling, including IL-13, and transforming growth factor beta (TGF-beta) by using gene-disrupted mice or treating mice with cytokine antagonists. We were surprised to find a high rate of rejection of osteosarcoma primary tumors in 88% (14 of 16) of CD1d KO mice compared with syngeneic wild-type BALB/c mice that showed rejection of tumor in <24% of mice. Further studies suggested that the rejection of tumor in CD1d KO mice was dependent on CD8(+) lymphocytes. Distinct from other murine tumor models, the negative regulation induced by CD1d-restricted NKT cells was not dependent on IL-4Ralpha-STAT6 signaling, including IL-13, or on TGF-beta. These data suggest that a novel CD1d-restricted NKT cell-mediated mechanism for tumor immunosuppression is active in the K7M2 osteosarcoma model and that NKT cells can regulate immunosurveillance through more than one pathway.


Role of Patient Age and Stone Density in Predicting Outcomes of Shockwave Lithotripsy in Lower Ureteral Stones.
Shockwave lithotripsy (SWL) is an important tool for treating ureteral stones, especially when ureteroscoy (URS) is not readily available. The aim of the study was to identify factors associated with the success of shockwave lithotripsy (SWL) in the treatment of lower ureteral stones. Total of 105 consecutive patients with single lower ureteral stones treated with SWL between January 2021 and February 2022 were reviewed retrospectively. Clinical and stone parameters including age, gender, and body mass index (BMI), stone length, and stone density were analyzed. SWL procedures were done using electro-conductive lithotripter Sonolith i-sys (EDAP TMS, Vaulx-en-Velin, France). We evaluated available clinical and radiologic features that might have been related to successful SWL treatment. Treatment was successful in 67/105 patients (63.8%). Multivariate logistic regression revealed that a successful outcome was significantly related to mean stone density (MSD) (95% CI: 1.004- 1.009) and the patients age (95% CI: 1.022- 1.13). Gender, BMI, and stone length were not an independent predicting factors. Stone density and patient age were independent predictors of successful outcome after two SWLs in the lower ureter. However, gender, BMI, and stone length did not independently predict treatment success.


Changes in myopia, visual acuity, and psychological distress after biofeedback visual training.
The effects of auditory biofeedback training on myopia, visual acuity (VA), and psychological distress were evaluated in a controlled prospective study involving 55 mildly myopic (< or = -3.5 D) high school students. These myopes were divided into 2 groups, matched for age and dioptric defect: 33 were treated with visual training and 22 were not; 27 emmetropic subjects formed a further control group. Subjects were evaluated at the baseline (T0), at 10 weeks after the end of the treatment (T1), and after an interval of 12 months (T2) from the baseline for: (1) manifest and cycloplegic refraction, and the difference between them (cycloplegic tonus); (2) VA measured with a conventional optotype in all subjects, and also with a computer-generated optotype in the treated group; and (3) psychometric values and personality profile. At T2, myopia had significantly worsened both in the treated and in the control myopes; VA in the treated myopes appeared significantly improved when measured by the conventional optotype, but unchanged when measured by computer. Psychometric scores improved significantly in the treated myopes and in the emmetropic controls. Objectively the autorefractometer showed that 38% of the myopes had voluntary control of positive accommodation, i.e., the ability to increase spherical defect; no voluntary control of negative accommodation was observed. An increase in VA was associated with a significant increase in the foveation time (i.e., the period of time when the target is imaged on the fovea and the eye is motionless), and was partly attributable to a learning effect. In conclusion, biofeedback visual training had a positive effect on psychological distress and subjective VA improvement, but failed to reduce the existing myopia or delay its evolution.


Factors associated with recurrent emergency department visits among people living with dementia: A retrospective cohort study.
Research on factors associated with recurrent emergency department (ED) visits and their implications for improving dementia care is lacking. The objective of this study was to examine associations between the individual characteristics of older adults living with dementia and recurrent ED visits. We used health administrative databases to conduct a population-based retrospective cohort study among older adults with dementia in Ontario, Canada. We included community-dwelling adults 66 years and older who visited the ED between April 1, 2010, and March 31, 2019 and were discharged home. We recorded all ED visits within one year after the baseline visit. We used recurrent event Cox regression to examine associations between repeat ED visits and individual clinical, demographic, and health service use characteristics. We fit conditional inference trees to identify the most important factors and define subgroups of varying risk. Our cohort included 175,863 older adults with dementia. ED use in the year prior to baseline had the strongest association with recurrent visits (3+ vs.0 adjusted hazard ratio (aHR): 1.92 (1.89, 1.94), 2vs.0 aHR: 1.45 (1.43, 1.47), 1vs.0 aHR: 1.23 (1.21, 1.24)). The conditional inference tree utilized history of ED visits and comorbidity count to define 12 subgroups with ED revisit rates ranging from 0.79 to 7.27 per year. Older adults in higher risk groups were more likely to live in rural and low-income areas and had higher use of anticonvulsants, antipsychotics, and benzodiazepines. History of ED visits may be a useful measure to identify older adults with dementia who would benefit from additional interventions and supports. A substantial proportion of older adults with dementia have a pattern of recurrent visits and may benefit from dementia-friendly and geriatric-focused EDs. Collaborative medication review in the ED and closer follow-up and engagement with community supports could improve patient care and experience.


Properties of permeation pathways induced in the human red cell membrane by malaria parasites.
The intracellular development of malaria parasites in mature erythrocytes imposes on the host cell a major demand for supply of nutrients and disposal of waste products. So as to cope with these demands, the erythrocyte membrane undergoes profound alterations in its basic permeability properties. A few hours after being invaded by Plasmodium falciparum parasites, and before any structural changes are apparent on the surface, the molecular traffic across the red cell membrane changes both in intensity and in composition of permeating substances. The changes are of a gradual nature, developmentally related and dependent on de novo protein synthesis, but do not occur concurrently for all the classes of permeants. Molecules which permeate very poorly into uninfected cells, such as hexitols (e.g., sorbitol and myoinositol), amino acids (e.g., glutamine, threonine, and histidine), a variety of organic acids and metal ions show a marked increase in their permeation rates across the host cell membrane. Likewise, substances whose normal permeation pathways conform with those of facilitated diffusion (e.g., hexoses, nucleosides, choline, and some amino acids), gain access into the host cytosol either by modified or additional permeation pathways. It has been proposed that three major new pathways are induced in the membrane of infected cells: (1) one of pore-like properties, which can accommodate most of the water soluble permeants, including anionic substances; (2) a protein-lipid interface, which can accommodate compounds of relatively higher hydrophobic character; and (3) modified constitutive transporters or modified lipid surroundings with altered transport activities. The pores are blocked by permeant bioflavonoid glycosides whose sites of binding are endofacial, and amount to less than a thousand per cell. In addition to serving as specific targets for transport blockers, the new sites of permeation can also serve as routes for enhanced delivery of cytotoxic agents into parasitized cells.


Relationship between estimated glomerular filtration rate and biological half-life of 131I. Retrospective analysis in patients with differentiated thyroid carcinoma.
This retrospective study sought to investigate the relationship between biological half-life (t1/2 biol) of 131I and estimated glomerular filtration rate (eGFR) in patients with thyroid carcinoma. 96 patients with differentiated thyroid carcinoma (69 women, 27 men, mean age 64.0 ± 13.6 years) and diagnostic and therapeutic administration of 131I were considered. Patients with pronounced specific iodine storage were not included in the study. The eGFR was estimated according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formula, the t1/2 biol via dosimetry. Patients were subdivided in groups with normal clearance (NC) (n = 37, 38.5%), medium clearance (MC) (n = 48, 50.0%), and low clearance (LC) (n = 11, 11.5%) (eGFR ≥ 90; 60-89; 15-59 ml/min per 1.73 m2, respectively). The relationship between eGFR and t1/2 biol of 131I was modeled using a power function. The groups significantly differed in terms of age (NC 53.8, MC 68.6, and 78.0 years, respectively), serum creatinine levels (NC: 0.71; MC: 0.85; LC: 1.18 mg/dl), and t1/2 biol (NC: 0.53; MC: 0.71; LC: 1.01 days). The t1/2 biol was significantly influenced only by eGFR, and not by age, gender, or body weight. The relationship between t1/2 biol of 131I and eGFR was described by the formula t1/2 biol = 20.3 · eGFR-0.782. The calculated relationship between renal function and t1/2 biol of 131I can be used in principle to estimate a dose reduction for patients with renal insufficiency. The model, however, gives erroneous results in individual cases and therefore a routine utilization cannot be recommended. Prospective studies are necessary, based on larger patient numbers and more accurate methods for dose rate measurement and GFR.


Characterization of protease-resistant fragments of laminin mediating attachment and spreading of rat hepatocytes.
In attempts to identify cell binding domains in the basement membrane protein laminin (Mr = 900,000), responsible for the substrate attachment mediating activity of the protein, proteolytic fragments were isolated and compared with respect to their biological activity. The fragments analyzed were generated by digestion of the protein with elastase or pepsin and included the previously described fragments 1 to 4 and three additional fragments, 5, 6, and 7 with molecular weights of 30,000 to 50,000. Fragments 1, 5, and 6 promoted substrate attachment of rat hepatocytes. Fragment 5, and to a lesser extent also fragment 6, but not fragment 1, induced spreading of the cells. Other fragments including the heparin-binding domain were inactive. Structural and immunological analyses indicated that fragment 1 is distinctly different from the other cell-binding fragments, whereas fragment 5 and 6 are similar but not identical. Furthermore, the active fragments were localized to different regions of the three short arms of the cross-shaped laminin molecule. Thus, the data suggest that fragments 1, 5, and 6 represent three separate domains with cell binding capacity. Attachment of hepatocytes to the fragments but not to intact laminin could be inhibited by specific antibodies indicating that the intact protein may contain an additional cell-binding site.


Molecular Characterization Revealed the Role of Thaumatin-Like Proteins of Bread Wheat in Stress Response.
Thaumatin-like proteins (TLPs) are related to pathogenesis-related-5 (PR-5) family and involved in stress response. Herein, a total of 93 TLP genes were identified in the genome of Triticum aestivum. Further, we identified 26, 27, 39, and 37 TLP genes in the Brachypodium distachyon, Oryza sativa, Sorghum bicolor, and Zea mays genomes for comparative characterization, respectively. They could be grouped into small and long TLPs with conserved thaumatin signature motif. Tightly clustered genes exhibited conserved gene and protein structure. The physicochemical analyses suggested significant differences between small and long TLPs. Evolutionary analyses suggested the role of duplication events and purifying selection in the expansion of the TLP gene family. Expression analyses revealed the possible roles of TLPs in plant development and abiotic and fungal stress response. Recombinant expression of TaTLP2-B in Saccharomyces cerevisiae provided significant tolerance against cold, heat, osmotic, and salt stresses. The results depicted the importance of TLPs in cereal crops that would be highly useful in future crop improvement programs.


Spike-associated and spike-independent esophageal contractions in patients with symptomatic diffuse esophageal spasm.
We report the electrical correlates of esophageal and lower esophageal sphincter contraction in humans using a bipolar ring electrode assembly mounted on a catheter. No spike activity was seen in the resting state in the esophagus or lower esophageal sphincter. Swallowing induced both electrical spike activity and contractile activity in both areas. The delay in the proximal and distal esophagus from swallowing to the onset of electrical activity was 0.3 +/- 0.1 s and 3.3 +/- 0.2 s, respectively, and from swallowing to the onset of contractile activity the time was 1.9 +/- 0.1 s and 5.2 +/- 0.2 s. respectively. The rate of propagation of electrical activity was constant throughout the esophageal body while contractile activity was propagated more rapidly in the middle third of the esophagus. Spike activity preceded the peak of esophageal contractions in 100% of swallows. Lower esophageal sphincter relaxation was initiated without spike activity, but the postswallow lower esophageal sphincter contraction was preceded by spike activity in 100% of cases. Basal lower esophageal sphincter pressure was maintained in the absence of spike activity. Spontaneous esophageal contractions, seen in patients with symptomatic diffuse esophageal spasm, were spike-independent in 70% of cases. Prolonged repetitive contractions, once initiated, were also spike-independent. Thus, with the use of intraluminal ring electrodes, we suggest that the human esophagus and lower esophageal sphincter are capable of generating both spike-dependent and spike-independent contractions. The latter may be prominent in patients with esophageal motor disorders.


Risk prediction of developing venous thrombosis in combined oral contraceptive users.
Venous thromboembolism (VTE) is a complex multifactorial disease influenced by genetic and environmental risk factors. An example for the latter is the regular use of combined oral contraceptives (CC), which increases the risk to develop VTE by 3 to 7 fold, depending on estrogen dosage and the type of progestin present in the pill. One out of 1'000 women using CC develops thrombosis, often with life-long consequences; a risk assessment is therefore necessary prior to such treatment. Currently known clinical risk factors associated with VTE development in general are routinely checked by medical doctors, however they are far from being sufficient for risk prediction, even when combined with genetic tests for Factor V Leiden and Factor II G20210A variants. Thus, clinical and notably genetic risk factors specific to the development of thrombosis associated with the use of CC in particular should be identified. Step-wise (logistic) model selection was applied to a population of 1622 women using CC, half of whom (794) had developed a thromboembolic event while using contraceptives. 46 polymorphisms and clinical parameters were tested in the model selection and a specific combination of 4 clinical risk factors and 9 polymorphisms were identified. Among the 9 polymorphisms, there are two novel genetic polymorphisms (rs1799853 and rs4379368) that had not been previously associated with the development of thromboembolic event. This new prediction model outperforms (AUC 0.71, 95% CI 0.69-0.74) previously published models for general thromboembolic events in a cross-validation setting. Further validation in independent populations should be envisaged. We identified two new genetic variants associated to VTE development, as well as a robust prediction model to assess the risk of thrombosis for women using combined oral contraceptives. This model outperforms current medical practice as well as previously published models and is the first model specific to CC use.


Morphologic and physiologic changes induced by Clostridium perfringens type A alpha toxin in the intestine of sheep.
To evaluate the morphologic and physiologic changes induced by Clostridium perfringens type A (alpha toxin in the ileum and colon of sheep. 16 ligated intestinal loops in 4 Merino lambs and 18 explants of ileum and colon from slaughtered lambs. alpha Toxin-induced fluid accumulation was evaluated in ligated ileal and colonic loops of sheep. Tissues were evaluated morphologically by use of gross and histologic examination. Effects of toxin on in vitro intestinal net water transport were tested in modified Ussing chambers. Ovine ileal and colonic loops incubated with C perfringens type A alpha toxin retained more fluid than control loops. Histologically, in the ileum of lambs inoculated with 300 LD50 of alpha toxin/mL, there was a mild to moderate multifocal infiltration of neutrophils in the lamina propria and submucosa. The colonic loops of lambs inoculated with 30 or 300 LD50 of alpha toxin/mL had excessive mucus in the lumen, a moderate amount of neutrophils mixed with mucus in the intestinal lumen, and moderate multifocal infiltration of the lamina propria and submucosa with neutrophils; the blood vessels of these layers were engorged with neutrophils. In vitro measurements of water transport also revealed inhibition of net epithelial water absorption in ileum and colon incubated with alpha toxin on the mucosal side. These results indicate that alpha toxin induces alterations in sheep intestine. Clostridium perfringens type A organisms that produce alpha toxin could be responsible for diseases of intestinal origin in some ruminants.


Isolation, Characterization, and Degradation Performance of the 17β-Estradiol-Degrading Bacterium Novosphingobium sp. E2S.
A 17β-estradiol (E2)-degrading bacterium E2S was isolated from the activated sludge in a sewage treatment plant (STP). The morphology, biological characteristics, and 16S ribosomal RNA (rRNA) gene sequence of strain E2S indicated that it belonged to the genus Novosphingobium. The optimal degrading conditions were 30 °C and pH 7.0. The ideal inoculum volume was 5% (v/v), and a 20-mL degradation system was sufficient to support the removal ability of strain E2S. The addition of extra NaCl to the system did not benefit the E2 degradation in batch culture by this strain. Strain E2S exhibited high degradation efficiency with initial substrate concentrations of 10-50 mg·L-1. For example, in mineral salt medium containing 50 mg·L-1 of E2, the degradation efficiency was 63.29% after seven days. In cow manure samples supplemented with 50 mg·L-1 of E2, strain E2S exhibited 66.40% degradation efficiency after seven days. The finding of the E2-degrading strain E2S provided a promising method for removing E2 from livestock manure in order to reduce the potential environmental risks of E2.


Measurement of directional thermal properties of biomaterials.
This paper presents an experimental technique to measure the directional thermal conductivity and thermal diffusivity of materials. A heated thermistor heats the sample and a sensing thermistor placed about 2.5 mm away measures the temperature rise due the heating pulse at the heated thermistor. An empirical relation between the power delivered by the first thermistor and the temperature rise recorded by the sensing thermistor is used to measure the thermal conductivity of the material along the line joining the thermistors. Diffusivity of the material is determined from the delay between the power pulse in the heated thermistor and the temperature pulse at the sensing thermistor. Signal processing was done to eliminate errors in the measurement due to change of base line temperature. Uncertainty of the measurement technique was found to be 5% when tested in media of known thermal properties. The thermal conductivity and thermal diffusivity of swine left ventricle in normal and ablated conditions were measured using this technique. The thermal conductivity of the tissue dropped significantly from 0.61 to 0.50 W.m(-1).K(-1) after ablation while the diffusivity dropped from 2.1 x 10(-7) to 1.7 x 10(-7)m2.s(-1).


Computational design of p-(dimethylamino)benzylidene-derived push-pull polyenes with high first-hyperpolarizabilities.
Multiple theoretical investigations on three new series of donor-bridge-acceptor substituted compounds are employed to aid in the design of NLO-phores with high first-hyperpolarizability β. The effect of varying the acceptor (rhodanine, thiohydantoin and thiobarbituric acid derivative-based) and bridge parts of these D-π-A systems was analyzed in terms of geometric and optoelectronic parameters such as bond length alternation, ground state dipole moments, HOMO and LUMO energies, UV-vis absorption spectra, transition dipole moments, and electronic absorption energies. Various functionals with the AUG-cc-pVDZ basis set including B3LYP, PBE38, and ωB97XD, and the Hartree-Fock method were employed to calculate β values, and the solvent effect was also considered by employing the SMD model. The variation of first-hyperpolarizabilities has been explained satisfactorily in terms of the PBE38/AUG-cc-pVDZ level calculated spectroscopic properties in the light of the sum-over-states method and the two-level model. The comprehensive study indicates that the most worthwhile targets for development as NLO-phores are compounds that include a longer π-bridge.


Zinc sulphate administered by transdermal iontophoresis improves breaking strength of surgical wounds in skin of alloxan-induced diabetic rats.
To investigate the effect of zinc sulphate administered by transdermal iontophoresis (TDI) on mechanical resistance of surgical wounds performed in the skin of diabetic rats. One hundred and sixty male Wistar rats weighing approximately 250 g were submitted to an incision surgery at the anterior region of abdomen and randomly distributed into four experimental groups with 40 non-diabetic control animals (G1) and 40 untreated diabetic animals (G2), both without any treatment of incisions; 40 non-diabetic animals (G3) and 40 untreated diabetic animals (G4), both with incisions treated with zinc sulphate, administered for a period of four consecutive days after surgery, in sessions of ten minutes duration, using a continuous-current electrostimulator (Zn + TDI). Each experimental group was further divided into four subgroups with ten rats each to be evaluated on the 4th, 7th, 14th, and 21st day after surgery. In each period were analyzed clinical and laboratory from the animals, and measured the breaking strength and hydroxyproline content (OH-P) of the skin scars. Breaking strength (BS) was significantly reduced (p<0.05) in skin scars of untreated diabetic rats (G2) on the 7th, 14th, and 21st postoperative days when compared to non-diabetic control rats (G1). In contrast, BS in skin scars of non-diabetic and untreated diabetic rats (G3, G4) treated with Zn + TDI showed significant increase (p<0.05) in those periods when compared with their respective controls with untreated incisions. The OH-P content of the scars did not show statistically significant variation in all studied groups at four different times evaluated after surgery. Zinc sulphate administered by transdermal iontophoresis had beneficial effect on the mechanical resistance of scars produced in the skin of diabetic rats. This therapeutic may have potential to reduce the complications observed in surgical wounds of the skin in diabetic subjects, mainly in most vulnerable stages of incisions to dehiscences, leakages and infections.


Audiovisual segregation in cochlear implant users.
It has traditionally been assumed that cochlear implant users de facto perform atypically in audiovisual tasks. However, a recent study that combined an auditory task with visual distractors suggests that only those cochlear implant users that are not proficient at recognizing speech sounds might show abnormal audiovisual interactions. The present study aims at reinforcing this notion by investigating the audiovisual segregation abilities of cochlear implant users in a visual task with auditory distractors. Speechreading was assessed in two groups of cochlear implant users (proficient and non-proficient at sound recognition), as well as in normal controls. A visual speech recognition task (i.e. speechreading) was administered either in silence or in combination with three types of auditory distractors: i) noise ii) reverse speech sound and iii) non-altered speech sound. Cochlear implant users proficient at speech recognition performed like normal controls in all conditions, whereas non-proficient users showed significantly different audiovisual segregation patterns in both speech conditions. These results confirm that normal-like audiovisual segregation is possible in highly skilled cochlear implant users and, consequently, that proficient and non-proficient CI users cannot be lumped into a single group. This important feature must be taken into account in further studies of audiovisual interactions in cochlear implant users.


Mitogen stimulation of Na+-H+ exchange: differential involvement of protein kinase C.
The mitogen-induced activation of Na+-H+ exchange was investigated in two cultured human fibroblast strains (HSWP and WI-38 cells) that, based on previous studies, differed in their response to the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) (L. M. Vincentini and M. L. Villereal, Proc. Natl. Acad. Sci. USA 82: 8053-8056, 1985). The role of protein kinase C in the activation of Na+-H+ exchange was investigated by comparing the effects of TPA on Na+ influx, in vitro phosphorylation, and in vivo phosphorylation in both cell types. Although both cell types have significant quantities of protein kinase C activity that can be activated by TPA in intact cells, the addition of TPA to intact cells stimulates Na+ influx in WI-38 cells but not in HSWP cells, indicating that in HSWP cells the stimulation of protein kinase C is not sufficient to activate the Na+-H+ exchanger. Cells were then depleted of protein kinase C activity by chronic treatment with high doses of TPA. Both HSWP and WI-38 cells were rendered protein kinase C deficient by this treatment as determined by in vitro and in vivo phosphorylation studies. Protein kinase C-deficient HSWP cells lose the ability for TPA to inhibit the serum-induced activation of Na+-H+ exchange, but there is no reduction in the stimulation of Na+ influx by serum, bradykinin, vasopressin, melittin, or vanadate, indicating that protein kinase C activity is not necessary for the mitogen-induced activation of Na+-H+ exchange in HSWP cells by agents known to stimulate phosphatidylinositol turnover (G. A. Jamieson and M. Villereal. Arch. Biochem. Biophys. 252: 478-486, 1987). In contrast, depletion of protein kinase C activity in WI-38 cells significantly reduces both the TPA- and the serum-induced activation of the Na+-H+ exchange system, suggesting that protein kinase C activity is necessary for at least a portion of the mitogen-induced activation of the Na+-H+ exchanger in WI-38 cells. These results indicate that the mechanisms for regulating Na+-H+ exchange can differ dramatically between different types of fibroblasts.


Implementation of a Novel Adherence Monitoring Strategy in a Phase III, Blinded, Placebo-Controlled, HIV-1 Prevention Clinical Trial.
Placebo-controlled HIV-1 prevention trials of pre-exposure prophylaxis (PrEP) have not generally used concurrent measurement of adherence because of the potential risk of unblinding. However, several pre-exposure prophylaxis trials for HIV-1 prevention among women failed to show effectiveness because of low product adherence. Evaluation of product adherence objectively during a study provides the opportunity for strengthening adherence activities at sites having low adherence. During MTN-020/ASPIRE, a phase III, placebo-controlled trial of the dapivirine intravaginal ring, we implemented an adherence monitoring system. Monitoring began in quarter 1 (Q1) 2013 and continued through the conclusion of the trial. Blood plasma was collected quarterly and tested for dapivirine concentrations while maintaining blinding among study team members involved in participant management. Dapivirine concentrations >95 pg/mL, reflecting >8 hours of continuous use, were assessed as signaling product use. Study leadership monitored results on a monthly basis and provided feedback to site investigators. Experiences were shared across sites to motivate staff and counsel participants to strive toward higher adherence levels. An upward trend in adherence was observed (P < 0.0001); the proportion of samples from subjects in the active arm with dapivirine >95 pg/mL increased from 63% in Q1 2013 to 84% by Q1 2015. Ongoing drug level testing as a marker of adherence in MTN-020/ASPIRE demonstrates the feasibility of real-time adherence monitoring while maintaining study blinding at the level of participants, sites, and study leadership. This approach is novel for large-scale effectiveness studies for HIV-1 prevention.


The enzyme-sensitive release of prodigiosin grafted β-cyclodextrin and chitosan magnetic nanoparticles as an anticancer drug delivery system: Synthesis, characterization and cytotoxicity studies.
In present investigation, two glucose based smart tumor-targeted drug delivery systems coupled with enzyme-sensitive release strategy are introduced. Magnetic nanoparticles (Fe3O4) were grafted with carboxymethyl chitosan (CS) and β-cyclodextrin (β-CD) as carriers. Prodigiosin (PG) was used as the model anti-tumor drug, targeting aggressive tumor cells. The morphology, properties and composition and grafting process were characterized by transmission electron microscope (TEM), Fourier transform infrared spectroscopy (FT-IR), vibration sample magnetometer (VSM), X-ray diffraction (XRD) analysis. The results revealed that the core crystal size of the nanoparticles synthesized were 14.2±2.1 and 9.8±1.4nm for β-CD and CS-MNPs respectively when measured using TEM; while dynamic light scattering (DLS) gave diameters of 121.1 and 38.2nm. The saturation magnetization (Ms) of bare magnetic nanoparticles is 50.10emucm-3, while modification with β-CD and CS gave values of 37.48 and 65.01emucm-3, respectively. The anticancer compound, prodigiosin (PG) was loaded into the NPs with an encapsulation efficiency of approximately 81% for the β-CD-MNPs, and 92% for the CS-MNPs. This translates to a drug loading capacity of 56.17 and 59.17mg/100mg MNPs, respectively. Measurement of in vitro release of prodigiosin from the loaded nanocarriers in the presence of the hydrolytic enzymes, alpha-amylase and chitosanase showed that 58.1 and 44.6% of the drug was released after one-hour of incubation. Cytotoxicity studies of PG-loaded nanocarriers on two cancer cell lines, MCF-7 and HepG2, and on a non-cancerous control, NIH/3T3 cells, revealed that the drug loaded nanoparticles had greater efficacy on the cancer cell lines. The selective index (SI) for free PG on MCF-7 and HepG2 cells was 1.54 and 4.42 respectively. This parameter was reduced for PG-loaded β-CD-MNPs to 1.27 and 1.85, while the SI for CS-MNPs improved considerably to 7.03 on MCF-7 cells. Complementary studies by fluorescence and confocal microscopy and flow cytometry confirm specific targeting of the nanocarriers to the cancer cells. The results suggest that CS-MNPs have higher potency and are better able to target the prodigiosin toxicity effect on cancerous cells than β-CD-MNPs.


Risk of developing seizure after percutaneous endoscopic lumbar discectomy.
A retrospective analysis in patients who underwent percutaneous endoscopic lumbar discectomy (PELD) and developed seizures during the procedure; and to identify the risk of developing seizure during PELD by measuring cervical epidural pressure. To evaluate clinical significance, characteristics, and risk factors for developing seizure and neck pain in patients undergoing PELD. Increased epidural pressure during PELD has been reported earlier. Risk of developing intraoperative seizure has not been investigated till date. We experienced some unexpected complication such as, seizures during PELD, and, therefore, we correlated it with the prodromal symptom and the strategies to avoid such complications during PELD. Four of the total 16,725 patients who underwent PELD between 2000 and 2008 developed intraoperative seizures. A review of their medical records and radiologic files were correlated with the complication. Factors evaluated were the type of seizures, prodromal symptoms, comorbidities and clinical outcome. To postulate a pathophysiologic cause of seizure, we designed a study to monitor the intraoperative cervical epidural pressure in 33 patients undergoing PELD. A striking feature of the 4 patients in this series was that they all complained of neck pain before the seizure event. There was no identifiable pattern of seizure observed. The duration of the procedure in these patients was longer than uninvolved cases. None of the patients developed any type of sequel subsequent to seizure. The outcome of surgery has been similar with the patients that did not have any type of complications after PELD. In the subsequent study of cervical epidural pressure, no patients developed seizure. However, there was occurrence of neck pain in the group with increased cervical epidural pressure. Although rare (0.02%), seizure can occur in patients undergoing PELD, occurrence of neck pain is correlated with increase in cervical epidural pressure, which should be considered as prodromal sign and alert the surgeon. Duration of procedure and speed of infusion are associated risk factor.


Possible biochemical impact of malaria infection in subjects with HIV co-infection in Anambra state, Nigeria.
The present study was designed to determine possible contributory impact of malaria infection on some biochemical markers in subjects with HIV co-infection in order to know if they are adverse or protective. Participants were recruited at the Voluntary Counseling and Testing Unit, Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria and grouped into: (i) Malaria and HIV co-infection group (n = 45); and (ii) HIV infected group without concurrent malaria infection (n = 57). Standard laboratory methods were used for the HIV and Plasmodium falciparum antigen screening, malaria parasite density, CD4+ T-cell count, packed cell volume, white blood cell count, serum iron and albumin concentrations. The results showed that serum iron and albumin were significantly reduced and raised respectively in 'Malaria-HIV co-infection group' compared with 'HIV infection group' (p < 0.05 and p < 0.05). A positive association was observed between age and serum iron concentration in malaria and HIV co-infected group (r = 0.580; p < 0.05) while negative associations were observed between PCV and serum iron (r = - 0.388; p < 0.05) and between CD4+ T-cells and serum iron concentration (r = -0.362; p < 0.05) in malaria and HIV co-infected group. The CD4+ T-cell count, WBC count, PCV were not significantly different between the Malaria-HIV co-infection group and HIV infection group. In the present study serum iron and albumin concentrations were the most sensitive indicators that showed the contributory impact of malaria infection on biochemical index in HIV co-infected subjects. The findings suggest that at the defined stage of HIV infection in the present study, malaria co-infection may moderate the impact of HIV infection on iron metabolism and hepatic synthesis of albumin.


Bacterial flocculation and production of poly- -hydroxybutyrate.
Experiments with a number of bacteria isolated from activated sludge have shown that flocculation is independent of the presence of poly-beta-hydroxybutyrate (PHB) in the cells. Several strains gave flocculent growth without any PHB detectable. Other strains, producing PHB in varying amounts, utilized this compound as an endogenous substrate, and after its disappearance the floc structure remained unchanged. The PHB content of various samples of activated sludge was found to be negligible.


Sex Differences in Compositional Plaque Volume Progression in Patients With Coronary Artery Disease.
This study sought to explore sex-based differences in total and compositional plaque volume (PV) progression. It is unclear whether sex has an impact on PV progression in patients with coronary artery disease (CAD). The study analyzed a prospective multinational registry of consecutive patients with suspected CAD who underwent 2 or more clinically indicated coronary computed tomography angiography (CTA) at ≥2-year intervals. Total and compositional PV at baseline and follow-up were quantitatively analyzed and normalized using the analyzed total vessel length. Multivariate linear regression models were constructed. Of the 1,255 patients included (median coronary CTA interval 3.8 years), 543 were women and 712 were men. Women were older (62 ± 9 years of age vs. 59 ± 9 years of age; p < 0.001) and had higher total cholesterol levels (195 ± 41 mg/dl vs. 187 ± 39 mg/dl; p = 0.002). Prevalence of hypertension, diabetes, and family history of CAD were not different (all p > 0.05). At baseline, men possessed greater total PV (31.3 mm3 [interquartile range (IQR): 0 to 121.8 mm3] vs. 56.7 mm3 [IQR: 6.8 to 152.1 mm3] p = 0.005), and there was an approximately 9-year delay in women in developing total PV than in men. The prevalence of high-risk plaques was greater in men than women (31% vs. 20%; p < 0.001). In multivariate analysis, after adjusting for age, clinical risk factors, medication use, and total PV at baseline, despite similar total PV progression rates, female sex was associated with greater calcified PV progression (β = 2.83; p = 0.004) but slower noncalcified PV progression (β = -3.39; p = 0.008) and less development of high-risk plaques (β = -0.18; p = 0.049) than in men. The compositional PV progression differed according to sex, suggesting that comprehensive plaque evaluation may contribute to further refining of risk stratification according to sex. (NCT02803411).


[Quadruple coronary endarterectomy while off-pump coronary artery bypass surgery - a case report].
Coronary endarterectomy is a controversial procedure. It has got many supporters and enemies among cardiac surgeons. It is very rare, particularly while performing off pump coronary artery bypass grafting. We present a case of a 50-year-old man who underwent coronary artery stenting and than has been admitted to surgery because of the restenosis in coronary arteries. During the surgery four arteries have been grafted. At the same time we performed four endarteriectomies in these vessels. No major problem was observed. We describe our technique and briefly present current literature regarding this problem.


Insecurity, distress and mental health: experimental and randomized controlled trials of a psychosocial intervention for youth affected by the Syrian crisis.
Strengthening the evidence base for humanitarian interventions that provide psychosocial support to war-affected youth is a key priority. We tested the impacts of an 8-week programme of structured activities informed by a profound stress attunement (PSA) framework (Advancing Adolescents), delivered in group-format to 12-18 year-olds in communities heavily affected by the Syrian crisis. We included both Syrian refugee and Jordanian youth. We followed an experimental design, comparing treatment youth and wait-list controls over two programme implementation cycles, randomizing to study arm in cycle 2 (ClinicalTrials.gov ID: NCT03012451). We measured insecurity, distress, mental health difficulties, prosocial behaviour and post-traumatic stress symptoms at three time-points: baseline (n = 817 youth; 55% Syrian, 43% female), postintervention (n = 463; 54% Syrian, 47% female), and follow-up (n = 212, 58% Syrian, 43% female). Regression models assessed: prospective intervention impacts, adjusting for baseline scores, trauma exposure, age, and gender; differential impacts across levels of trauma exposure and activity-based modality; and sustained recovery 1 year later. We analysed cycle-specific and cycle-pooled data for youth exclusively engaged in Advancing Adolescents and for the intent-to-treat sample. We found medium to small effect sizes for all psychosocial outcomes, namely Human Insecurity (β = -7.04 (95% CI: -10.90, -3.17), Cohen's d = -0.4), Human Distress (β = -5.78 (-9.02, -2.54), d = -0.3), and Perceived Stress (β = -1.92 (-3.05, -0.79), d = -0.3); and two secondary mental health outcomes (AYMH: β = -3.35 (-4.68, -2.02), d = -0.4; SDQ: β = -1.46 (-2.42, -0.50), d = -0.2). We found no programme impacts for prosocial behaviour or post-traumatic stress reactions. Beneficial impacts were stronger for youth with exposure to four trauma events or more. While symptoms alleviated for both intervention and control groups over time, there were sustained effects of the intervention on Human Insecurity. Findings strengthen the evidence base for mental health and psychosocial programming for a generation affected by conflict and forced displacement. We discuss implications for programme implementation and evaluation research.


Natural selection in action during speciation.
The role of natural selection in speciation, first described by Darwin, has finally been widely accepted. Yet, the nature and time course of the genetic changes that result in speciation remain mysterious. To date, genetic analyses of speciation have focused almost exclusively on retrospective analyses of reproductive isolation between species or subspecies and on hybrid sterility or inviability rather than on ecologically based barriers to gene flow. However, if we are to fully understand the origin of species, we must analyze the process from additional vantage points. By studying the genetic causes of partial reproductive isolation between specialized ecological races, early barriers to gene flow can be identified before they become confounded with other species differences. This population-level approach can reveal patterns that become invisible over time, such as the mosaic nature of the genome early in speciation. Under divergent selection in sympatry, the genomes of incipient species become temporary genetic mosaics in which ecologically important genomic regions resist gene exchange, even as gene flow continues over most of the genome. Analysis of such mosaic genomes suggests that surprisingly large genomic regions around divergently selected quantitative trait loci can be protected from interrace recombination by "divergence hitchhiking." Here, I describe the formation of the genetic mosaic during early ecological speciation, consider the establishment, effects, and transitory nature of divergence hitchhiking around key ecologically important genes, and describe a 2-stage model for genetic divergence during ecological speciation with gene flow.


Changes in intercellular electrical coupling of smooth muscle accompanying atrophy and hypertrophy.
Longitudinal tissue impedance was determined for cat circular intestinal muscle that was either hypertrophied due to volume overloading or atrophied due to defunctionalization. These conditions were produced by bypassing 50 cm of jejuno-ileum in six cats and, 2-6 mo later, removing segments from the proximal jejunum of the hypertrophied functional gut and from the atrophied proximal end of the bypassed loop. Impedances were compared with those of jejunal circular muscle from 15 normal cats. Specific tissue impedance was determined by a modification of the method of Tomita (J. Physiol. Lond. 201: 145-159, 1969), which employs Krebs and Krebs-sucrose solutions; a tissue shrinkage of 5%, empirically found to occur in Krebs-sucrose solution, was corrected for. Impedance values were determined at 20 frequencies between 30 Hz and 30 kHz. The value at 30 kHz was taken to represent the specific myoplasmic resistance (Rmyo) of each tissue, while the difference between the value of 30 Hz and 30 kHz was taken to represent the specific junctional resistance (Rj). Values (in omega X cm) for Rmyo were control 134 +/- 2, functional 128 +/- 5, bypassed 151 +/- 6 (mean of means +/- SE). Corresponding values for Rj were control 173 +/- 15, functional 96 +/- 27, bypassed 340 +/- 75. Calculated values (in microF/cm) for junctional capacitance were control 2.66, functional 6.10, bypassed 1.97. Acid uncoupling by saturating the bathing solutions with 100% CO2 revealed a pH-sensitive resistive component of Rj, assumed to be attributable to gap junctions.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantitative reversed-phase high performance liquid chromatographic determination of major and modified deoxyribonucleosides in DNA.
We have developed a method to accurately determine (< 3% RSD) the complete major and modified base composition of a few micrograms of unlabeled DNA. The DNA samples were quantitatively hydrolyzed with DNase 1, Nuclease P1, and bacterial alkaline phosphatase. The resulting deoxyribonucleosides were directly separated in 70 min by reversed-phase high performance liquid chromatography with detection by ultraviolet absorption at 254 nm and 280 nm (RP-HPLC). The highly sensitive and selective dual wavelength quantitation greatly enhances the precision and accuracy of the chromatographic analysis. Contamination of DNA preparations with RNA does not interfere with the DNA analysis due to the high resolution of the chromatography. We have used this method for the quantitation of m5dCyd in 5 microgram of calf thymus and salmon sperm DNA in which the m5dCyd comprises only 1 to 2% of the total bases. This method should be a useful research tool in studies on various DNAs and DNA subfractions and should help to elucidate the functions of methylation of DNA.


Raman spectroscopy offers great potential for the nondestructive confirmatory identification of body fluids.
Raman spectroscopy was used to compare body fluids commonly found at crime scenes in a nondestructive manner. The dry traces of semen, vaginal fluid, sweat, saliva, and blood were analyzed using confocal Raman microscopy with a 785-nm excitation. The results show that the five fluids can be differentiated from one another by visual comparison of their Raman spectra, and that the laser radiation does not damage the sample. The Raman signature of each body fluid is specific and correlates with the known composition of the fluid. Dry traces of human and canine semen exhibited distinctly different Raman signatures. Overall, this preliminary study demonstrates the great potential of Raman spectroscopy for nondestructive, confirmatory identification of body fluids for forensic purposes.