Differences in the enzymatic efficiency of human and bony fish AID are mediated by a single residue in the C terminus modulating single-stranded DNA binding.
Activation-induced cytidine deaminase (AID) mediates antibody diversification by deaminating deoxycytidines to deoxyuridine within immunoglobulin genes. However, it also generates genome-wide DNA lesions, leading to transformation. Though the biochemical properties of AID have been described, its 3-dimensional structure has not been determined. Hence, to investigate the relationship between the primary structure and biochemical characteristics of AID, we compared the properties of human and bony fish AID, since these are most divergent in amino acid sequence. We show that AIDs of various species have different catalytic rates that are thermosensitive and optimal at native physiological temperatures. Zebrafish AID is severalfold more catalytically robust than human AID, while catfish AID is least active. This disparity is mediated by a single amino acid difference in the C terminus. Using functional assays supported by models of AID core and surface structure, we show that this residue modulates activity by affecting ssDNA binding. Furthermore, the cold-adapted catalytic rates of fish AID result from increased ssDNA binding affinity at lower temperatures. Our work suggests that AID may generate DNA damage with variable efficiencies in different organisms, identifies residues critical in regulating AID activity, and provides insights into the evolution of the APOBEC family of enzymes.


An efficient framework for automated screening of Clinically Significant Macular Edema.
The present study proposes a new approach to automated screening of Clinically Significant Macular Edema (CSME) and addresses two major challenges associated with such screenings, i.e., exudate segmentation and imbalanced datasets. The proposed approach replaces the conventional exudate segmentation based feature extraction by combining a pre-trained deep neural network with meta-heuristic feature selection. A feature space over-sampling technique is being used to overcome the effects of skewed datasets and the screening is accomplished by a k-NN based classifier. The role of each data-processing step (e.g., class balancing, feature selection) and the effects of limiting the region of interest to fovea on the classification performance are critically analyzed. Finally, the selection and implication of operating points on Receiver Operating Characteristic curve are discussed. The results of this study convincingly demonstrate that by following these fundamental practices of machine learning, a basic k-NN based classifier could effectively accomplish the CSME screening.


Imposex in Bolinus brandaris from the Ria formosa lagoon (southern Portugal): usefulness of "single-site baselines" for environmental monitoring.
The present study aimed at creating a so called "single-site baseline" (SSB) of imposex in the purple dye murex (Bolinus brandaris) from the Ria Formosa lagoon (Algarve coast, southern Portugal), at the moment that the IMO Antifouling System Convention (banning globally the use of organotin compounds in antifouling paints on ships' hulls) entered into force (September 2008). In the context of monitoring long-term temporal trends of pollution levels, SSBs are proposed whenever it is difficult to obtain adequate numbers of bioindicator specimens in most of the study area or if significant short-term variability in imposex levels is likely to occur. In order to deal with these constraints, this study created a SSB by sampling on a monthly-basis (from October 2008 to September 2009) in a single site considered to be representative of the Ria Formosa as a whole (vicinities of the Culatra Island) and where B. brandaris is abundant all year round. The SSB provided a good characterisation of the monthly variation in the imposex indices, which ranged throughout the year between 72.7 < I% < 100; 0.08 < FPLI < 1.73; 1.20 < RPLI < 23.27; 1.29 < VDSI < 3.54. Only a SSB could have detected this large variability, which otherwise might have been unnoticed and would have produced biased temporal and spatial comparisons, leading to erroneous interpretations of environmental monitoring data. Therefore, this SSB of imposex in B. brandaris constitutes valuable reference data to assess the future trend of organotin pollution in the Ria Formosa, after the benchmark date of the complete eradication of organotin from antifouling agents was implemented worldwide.


Acute renal failure in pregnancy: Tertiary centre experience from north Indian population.
Obstetrical acute renal failure ARF is now a rare entity in the developed countries but still a common occurrence in developing countries. Delay in the diagnosis and late referral is associated with increased mortality. This study aimed to evaluate the contributing factors responsible for pregnancy-related acute kidney failure, its relation with mortality and morbidity and outcome measures in these patients. Total 520 patients of ARF of various aetiology were admitted, out of these 60 (11.5%) patients were pregnancy-related acute renal failure. ARF Acute renal failure occurred in 32 (53.3%) cases in early part of their pregnancy, whereas in 28 (46.7%) cases in later of the pregnancy. Thirty-two (53.3%) patients had not received any antenatal visit, and had home delivery, 20 (33.4%) patients had delivered in hospitals but without antenatal care and eight (13.3%) patients received antenatal care and delivered in the hospitals. Anuria was observed in 23 (38.3%) cases, remaining 37 (61.7%) cases presented with oliguria. Septicemia was present in 25 (41.7%), hypertensive disorder of pregnancy in 20 (33.3%), haemorrhage in eight (13.3%), abortion in 5 (8.3%), haemolysis elevated liver enzymes low platelets counts (HELLP) syndrome in one (1.67%) and disseminated intravascular coagulation in one (1.67%). (61.7%) patients were not dialyzed, 33 (55%) recovered normal renal function with conservative treatment. Complete recovery was observed in 45 (75%) patients, five (8.4%) patients developed irreversible renal failure. Maternal mortality was nine (15%) and foetal loss was 25 (41.7%). Pregnancy-related ARF is usually a consequence of obstetric complications; it carries very high morbidity and mortality.


Growth of Salmonella Typhimurium DT104 at 30°C is not affected by anatomical location on the chicken carcass.
Development of models for growth of Salmonella in the chicken food matrix is time-consuming and expensive. The current study was undertaken to examine growth of Salmonella on different anatomical locations of the chicken carcass. The purpose was to determine whether anatomical location should be included as an independent variable in predictive models for chicken. Eleven anatomical locations were studied: skin (wing, breast, drumstick, and thigh), meat surface (wing, breast, drumstick, and thigh), and meat interior (breast, drumstick, and thigh). Background microflora, pH, and growth (lag time, λ; growth rate, μ; and time for a 3-log increase, t(3)) at 30°C for a small inoculum size (0.92 ± 0.30 log per portion) of Salmonella Typhimurium DT104 were examined. Four or six replicate storage trials were conducted per anatomical location (n = 46 growth curves). Portion sizes were 1.12 ± 0.17 g (mean ± standard deviation) for meat and 0.25 ± 0.08 g for skin. A two-phase linear model was used to determine λ and μ. The effect of anatomical location on dependent variables was assessed by one-way analysis of variance. pH values differed (P < 0.001) among anatomical locations, with skin (6.86 ± 0.20). dark meat (6.39 ± 0.20) . white meat (5.97 ± 0.20). Background microflora (4.32 ± 1.66 log per portion) was variable and not affected (P > 0.05) by anatomical location. Likewise, λ (1.90 ± 0.75 h), μ (0.648 ± 0.120 log/h), and t(3) (6.71 ± 0.82 h) at 30°C were not affected (P > 0.05) by anatomical location. Although there were differences in pH among anatomical locations, these differences were not sufficient to affect growth of Salmonella Typhimurium DT104 at 30°C. If this observation holds for other storage conditions and strains, then anatomical location does not need to be included as an independent variable in predictive models for chicken. This would save significant time and money for the predictive microbiologist.


A novel ultraviolet assay for testing side reactions of carbodiimides.
Carbodiimides possess considerable absorbance in the ultraviolet region; the extinction coefficient of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) in water is epsilon(214) = 6.3. 10(3) L x mol(-1) x cm(-1). It provides a very simple method for testing possible side reactions of carbodiimides. This technique was used to study effects of pH, different buffers, and other components typically present in biological samples on EDC stability. It was shown that the hydrolysis rate in pure water increased from 1.5. 10(-5) to 5. 10(-4) s(-1) by pH decreasing from 7 to 4. A strong increase of the rate of EDC loss was observed in the presence of different components. This effect can be described by the following row of hydrolysis rates: citrate >> acetate approximately phosphate >> SDS. The results can be used to optimize carbodiimide-mediated reactions of peptide bond formation in organic chemistry or peptide synthesis or during immobilization or cross-linking of biological molecules.


Pantoea peritonitis in a patient receiving chronic ambulatory peritoneal dialysis.
Pantoea agglomerans is usually the most common organism transmitted through plant thorn injuries. This report is of a female patient maintained on chronic ambulatory peritoneal dialysis (CAPD) who developed peritonitis attributed to P. agglomerans. Peritonitis is an uncommon complication of P. agglomerans and there is no previous report of peritonitis associated with this organism in a CAPD patient. The source of infection was thought to be due to rose-thorn injury. Antibiotic therapy with ceftazidime and amikacin i.p. led to a clinical improvement, with disappearance of the organism in the peritoneal fluid.


Sources of Carbon for Export from Spinach Leaves throughout the Day.
Rates of net carbon exchange, export, starch, and sucrose synthesis were measured in leaves of spinach (Spinacia oleracea L.) throughout a 14-hour period of sinusoidal light to determine the sources of carbon contributing to export. Net carbon exchange rate closely followed light level, but export remained relatively constant throughout the day. In the morning when photosynthesis was low, starch degradation provided most of the carbon for export, while accumulated sucrose was exported during the evening. At high photosynthesis rate, the regulatory metabolite fructose 2,6-bisphosphate was low, allowing more of the newly fixed carbon to flow to sucrose through cytosolic fructose bisphosphatase. When the rate of sucrose synthesis exceeded the rate of export from the leaf, sucrose accumulated and soon thereafter sucrose synthesis declined. A decreasing sucrose synthesis rate resulted in additional carbon moving to the synthesis of starch, which was maintained throughout the remainder of the day. The declining sucrose synthesis rate coincided with decreasing activity of sucrose phosphate synthase present in gel-filtered leaf extracts. A rise in the leaf levels of uridine diphosphoglucose and fructose 6-phosphate throughout the day was consistent with this declining activity.


Development and Validation of Multi-Residue Method for Drugs Analysis in Human Feces by Liquid Chromatography-Tandem Mass Spectrometry.
The use of veterinary drugs in animal production is a common practice to secure animal and human health. However, residues of administrated drugs could be present in animal food products. Levels of drugs in food of animal origin are regulated within the European Union. In recent years, residues have been detected not only in food, but also in the environmental elements such as water or soil, meaning that humans are involuntarily exposed to these substances. This article presents a multiclass method for the analysis of various therapeutic groups of pharmaceuticals in human feces. Pharmaceuticals are extracted from feces with an acid extraction solvent, and after filtration the extract was analyzed by HPLC-MS/MS. A limit of detection of 10 ng/g was achieved for 9 pharmaceuticals, with linearity over 0.99 and repeatability and reproducibility lower than 20%. The method was satisfactorily applied in 25 feces samples of individuals that had declared not to be under medical treatment for the last two months. Results indicate the presence of six different compounds at concentration between 10 and 456 ng/g. This preliminary study showed the involuntary exposure of human gut microbiota to active substances such as pharmaceuticals.


Ab Initio Investigation of the Ground States of F2P(S)N, F2PNS, and F2PSN.
A recent spectroscopic experiment identified difluorothiophosphoryl nitrene (F2P(S)N) and found that it showed rich photochemistry. However, a discrepancy between the experimental results and the quantum chemical calculations was reported. Thus, high-level ab initio calculations using the coupled cluster singles and doubles with perturbative triples and second-order multiconfigurational perturbation theory were performed to elucidate this inconsistency. The discrepancy arose due to the failure to consider the triplet state of difluoro(thionitroso)phosphine (F2PNS). In this work, we identify that the global minimum of the system is the triplet state of F2PNS, which allows us to explain the inconsistency between the experimental and theoretical results. All calculated results give consistent results with the recent experimental results.


Sex steroid blockade enhances thymopoiesis by modulating Notch signaling.
Paradoxical to its importance for generating a diverse T cell repertoire, thymic function progressively declines throughout life. This process has been at least partially attributed to the effects of sex steroids, and their removal promotes enhanced thymopoiesis and recovery from immune injury. We show that one mechanism by which sex steroids influence thymopoiesis is through direct inhibition in cortical thymic epithelial cells (cTECs) of Delta-like 4 (Dll4), a Notch ligand crucial for the commitment and differentiation of T cell progenitors in a dose-dependent manner. Consistent with this, sex steroid ablation (SSA) led to increased expression of Dll4 and its downstream targets. Importantly, SSA induced by luteinizing hormone-releasing hormone (LHRH) receptor antagonism bypassed the surge in sex steroids caused by LHRH agonists, the gold standard for clinical ablation of sex steroids, thereby facilitating increased Dll4 expression and more rapid promotion of thymopoiesis. Collectively, these findings not only reveal a novel mechanism underlying improved thymic regeneration upon SSA but also offer an improved clinical strategy for successfully boosting immune function.


Structural and biochemical insights into the allosteric activation mechanism of AMP-activated protein kinase.
The AMP-activated protein kinase (AMPK), a complicated αβγ heterotrimer, can sense cellular energy status and maintain energy homeostasis via switching catabolic and anabolic pathways. AMPK also participates in the regulation of many other life activities, including the cell cycle, cell polarity, autophagy, and life span. Therefore, AMPK is widely studied as a potential drug target for treatment of type 2 diabetes and some other metabolic diseases, cancers, and cardiovascular diseases. Similar to other kinases, the phosphorylation of α-Thr172 in the activation loop by upstream kinases is crucial for the activation of AMPK. In addition, the binding of AMP and its analogues to the γ subunit causes further allosteric activation, which makes AMPK distinctive from other kinases. Here, we give a brief introduction to the domain constitutions of mammalian AMPK and then systematically review its allosteric activation mechanism from a structural and biochemical view.


Ecosystem Services, Global Diversity, and Rate of Stonefly Species Descriptions (Insecta: Plecoptera).
Stoneflies (Insecta: Plecoptera) provide ecosystem services as indicators of water quality, as food for predators, as mediators of energy flow and nutrient cycling, and through cultural services related to recreation and artistic creativity. The Plecoptera Species File (PSF) aggregates stonefly nomenclature, distribution, and literature to help society and scientists understand the value of services stoneflies provide. Using PSF data, we examined global and regional diversity, compared species description rates, and predicted future species description numbers through the year 2100. Through 2018, extant species totaled 3,718 with Temperate Asia having the greatest regional diversity at 1178 species. The Perlidae was the most species-rich of the 16 families at 1120 species. The recent global rate of species description was 43.6 species/yr, with Temperate Asia having the highest regional rate at 13.7 species/yr, followed by China and South America adding approximately 9.0 species/yr. We predicted that 1140 ± 130 new species would be described globally by 2050, and 2130 ± 330 by the year 2100, most of the increase occurring in China and South America. We discuss the possibility of reaching these predicted values.


Inhibition of collar-induced carotid atherosclerosis by recombinant apoA-I cysteine mutants in apoE-deficient mice.
The previous studies in our laboratory revealed that seven cysteine mutants of apolipoprotein A-I (apoA-I) have different structural features and biological activities in vitro and in vivo. To investigate the potential cardioprotective effects of apolipoprotein A-I(N74C) [apoA-I(N74C)], we examined the anti-inflammatory, antioxidant, and antiatherosclerotic effects of this cysteine mutant in a rapid atherosclerosis model induced by perivascular carotid collar placement in apoE⁻/⁻ mice. Lipid-free apoA-I(N74C) showed a significant increased antioxidant potency in low density lipoprotein (LDL) oxidation in vitro and reduced intracellular lipid accumulation in THP-1-derived macrophages, relative to wild-type apoA-I (apoA-Iwt). Mice injected with recombinant HDL (rHDL) reconstituted with apoA-I(N74C) (named rHDL74) through tail veins (40 mg/kg of body weight, three injections) had a significant lower level of serum interleukin-6 (IL-6) and enhanced serum antioxidation compared with mice receiving rHDL reconstituted with apoA-Iwt (named rHDLwt). Moreover, compared with rHDLwt, the rHDL74 in vivo injection resulted in a significant decrease in plaque size, ratio of aorta intima to media, arterial remodeling, and macrophage content in lesions. In summary, intravenous injection with rHDL74 reconstituted with apoA-I cysteine mutant apoA-I (N74C) dramatically delays the development of atherosclerosis induced by perivascular carotid collar placement and reduces vascular remodeling in the carotid artery in apoE⁻/⁻ mice.


Modeling the influence of motor skills on literacy in third grade: Contributions of executive functions and handwriting.
Several nonlanguage factors influence literacy development, and motor skills are among those most studied. Despite the publication of several studies that have supported the existence of this relationship, the type of influence and underlying mechanisms have been little explored. Herein, we propose modeling the relationship between motor skills and literacy through structural equation modeling, testing the contribution of executive functions and handwriting skills as the possible mediators of this relationship. In a study of 278 third-grade children, we used a wide range of measures related to written language (reading, spelling, reading comprehension, and written production), fine motor skills (dominant hand, nondominant hand, and bimanual dexterity), executive functions (verbal and visuospatial working memory, inhibition, and shifting), and handwriting. Structural equation modeling of the relationship between these different variables indicated that in the third grade, the influence of fine motor skills on literacy is fully mediated by both executive functions and handwriting skills. These motor skills effects are observed for both low levels of processing (reading, spelling) and high levels of processing (reading comprehension, written production). The results are discussed in terms of the potential mechanisms underlying different literacy skills and their implications for pedagogical programs.


[The overexpression of NCAM (CD56) in human hearts is specific for ischemic damage].
The main reason for myocardial dysfunction is chronical myocardial ischemia. Recently we could show, that NCAM (CD56), a neural cell adhesion molecule and member of the immunoglobuline superfamily, and the transcription factor AML1 (RUNX1) are overexpressed in chronic ischemic human heart failure compaired to normal hearts. Here we demonstrate, that the overexpression of NCAM (CD56) is specific for ischemic damage as compaired to other heart diseases including congestive cardiomyopathy, hypertrophic obstrutive cardiomyopathy, myocarditis and sarcoidosis. Concerning the transcriptional regulation of NCAM (CD56) by AML1 (RUNX1) we isolated 3 novel isoforms of AML 1 (RUNX1) with different transactivating function, that might be a regulatory element of the NCAM (CD56) overexpression in chronical myocardial ischemia.


Genetic variation of DNA methyltransferase-3A contributes to protection against persistent MRSA bacteremia in patients.
The role of the host in development of persistent methicillin-resistant Staphylococcus aureus (MRSA) bacteremia is not well understood. A cohort of prospectively enrolled patients with persistent methicillin-resistant S. aureus bacteremia (PB) and resolving methicillin-resistant S. aureus bacteremia (RB) matched by sex, age, race, hemodialysis status, diabetes mellitus, and presence of implantable medical device was studied to gain insights into this question. One heterozygous g.25498283A > C polymorphism located in the DNMT3A intronic region of chromosome 2p with no impact in messenger RNA (mRNA) expression was more common in RB (21 of 34, 61.8%) than PB (3 of 34, 8.8%) patients (P = 7.8 × 10-6). Patients with MRSA bacteremia and g.25498283A > C genotype exhibited significantly higher levels of methylation in gene-regulatory CpG island regions (Δmethylation = 4.1%, P < 0.0001) and significantly lower serum levels of interleukin-10 (IL-10) than patients with MRSA bacteremia without DNMT3A mutation (A/C: 9.7038 pg/mL vs. A/A: 52.9898 pg/mL; P = 0.0042). Expression of DNMT3A was significantly suppressed in patients with S. aureus bacteremia and in S. aureus-challenged primary human macrophages. Small interfering RNA (siRNA) silencing of DNMT3A expression in human macrophages caused increased IL-10 response upon S. aureus stimulation. Treating macrophages with methylation inhibitor 5-Aza-2'-deoxycytidine resulted in increased levels of IL-10 when challenged with S. aureus In the murine sepsis model, methylation inhibition increased susceptibility to S. aureus These findings indicate that g.25498283A > C genotype within DNMT3A contributes to increased capacity to resolve MRSA bacteremia, potentially through a mechanism involving increased methylation of gene-regulatory regions and reduced levels of antiinflammatory cytokine IL-10.


The abrasive effect of a porcelain and a nickel-chromium alloy on the wear of human enamel and the influence of a carbonated beverage on the rate of wear.
This study was conducted to determine the abrasive effect of a porcelain and an Ni-Cr alloy on the wear of human enamel, and the influence of a carbonated beverage on the rate of wear. Tooth specimens were prepared by embedding 48 freshly extracted mandibular first premolars in acrylic. Twenty-four of these specimens were abraded against Ni-Cr, and the remaining 24 against porcelain in artificial saliva and carbonated beverage media, respectively (n = 12), on a specially designed abrasive testing machine at a constant load of 40 N with 6 mm amplitude for 15,000 cycles. The cusp heights of the tooth specimens were measured both before and after abrasion using a profile projector. The abraded cast specimens were subjected to profilometry for computing the surface roughness; the abrading media was subjected to atomic absorption spectrophotometry for analyzing Ni and Cr ion levels. Data obtained were statistically analyzed. Porcelain specimens in a medium of carbonated beverage caused the highest wear of tooth specimens. The lowest wear of tooth specimens was Ni-Cr specimens in artificial saliva medium. Carbonated beverage caused significantly higher wear of tooth specimens when abraded against Ni-Cr and porcelain specimens than did artificial saliva. The mean quantitative surface roughness of porcelain specimens was significantly higher than that of Ni-Cr specimens, irrespective of the medium in which abrasion testing was conducted. There was no statistically significant difference between the concentrations of Ni ions released in artificial saliva and carbonated beverage media. Also, there was no statistically significant difference between the concentrations of Cr ions released in artificial saliva and carbonated beverage media. The wear of human enamel was significantly higher in the presence of carbonated beverage than artificial saliva and against porcelain when compared with Ni-Cr. The surface roughness of porcelain in the presence of carbonated beverage was found to be highest, and the release of Ni and Cr was not affected by carbonated beverage.


Complex, Isolated Pulmonary Arteriovenous Malformation in Two Children With Severe Cyanosis.
We present two cases of Isolated complex pulmonary arteriovenous malformations in two children presenting with cyanosis and exercise intolerance. We present the anatomical features and the management of each case. One patient was treated with surgical ligation of the draining vein, and one patient was treated percutaneously by closure of the feeding segmental artery.


Managing risk and exerting control: determining follow through with falls prevention.
This study used in-depth interviews to explore the perspectives of nine older women who had not followed through with environmental modification recommendations to reduce their risk of falls in the home. It was found that the core concept of 'exerting control' provided an understanding of their experience following an occupational therapy home visit. Exerting control was a behavioural, cognitive and affective process whereby the women made decisions about whether or not to follow through with environmental modification recommendations based on their knowledge of environmental risks, perceptions of degree of risk, perceived ability to mediate these risks through behaviour and the degree of freedom she had in decision making. Exerting control meant that the women made daily choices about their home environment which increased or decreased the risk of falls with identified home hazards. The findings suggest that, for some women, health professionals need to understand and work with the phenomenon of exerting control, in order to work with clients to reduce environmental hazards.


Neuropeptides in skin disease: increased VIP in eczema and psoriasis but not axillary hyperhidrosis.
The neuropeptides vasoactive intestinal polypeptide (VIP), substance P and somatostatin were studied in skin biopsies from patients with eczema, psoriasis and axillary hyperhidrosis. VIP concentrations were elevated in skin affected by eczema and psoriasis, whereas substance P and somatostatin levels did not differ from controls. There was a higher concentration of VIP, but not of substance P or somatostatin, in normal axillary skin when compared to adjacent trunk skin, with abundant VIP-containing fibres surrounding eccrine sweat glands. The VIP concentration was unchanged in skin affected by axillary hyperhidrosis. VIP may increase local blood flow in eczema and psoriasis, but does not appear to play a role in axillary hyperhidrosis.


Recent progress in plant-derived polysaccharides with prebiotic potential for intestinal health by targeting gut microbiota: a review.
Natural products of plant origin are of high interest and widely used, especially in the food industry, due to their low toxicity and wide range of bioactive properties. Compared to other plant components, the safety of polysaccharides has been generally recognized. As dietary fibers, plant-derived polysaccharides are mostly degraded in the intestine by polysaccharide-degrading enzymes secreted by gut microbiota, and have potential prebiotic activity in both non-disease and disease states, which should not be overlooked, especially in terms of their involvement in the treatment of intestinal diseases and the promotion of intestinal health. This review elucidates the regulatory effects of plant-derived polysaccharides on gut microbiota and summarizes the mechanisms involved in targeting gut microbiota for the treatment of intestinal diseases. Further, the structure-activity relationships between different structural types of plant-derived polysaccharides and the occurrence of their prebiotic activity are further explored. Finally, the practical applications of plant-derived polysaccharides in food production and food packaging are summarized and discussed, providing important references for expanding the application of plant-derived polysaccharides in the food industry or developing functional dietary supplements.


[Ropivacaine 0.2% versus bupivacaine 0.125% plus sufentanil for continuous peridural analgesia following extended abdominal operations].
An improved opioid-free analgesia concept using thoracic epidural anlgesia with a low concentrated local anaesthetic alone could be of practical interest for hospitals which are unable to guarantee continuous monitoring on the wards. We have compared in a prospective, randomized, double-blind study, the analgesic effects and side effects of ropivacaine 0.2% (Group 1) with bupivacaine 0.125% plus 0.5 microgram ml-1 sufentanil (Group 2) via a thoracic epidural catheter during the first 24 hours after major abdominal surgery in 28 gynaecological tumour patients. Assessments were performed 8, 12, 16, 20 and 24 hours after surgery. No differences were seen in duration of surgery, intraoperative opioid and supplemental analgesic requirements. As a main result VAS values for pain during mobilisation were higher in Group 1 when compared with Group 2 (p < 0.01) after 20 and 24 hours of continuous epidural infusion (10 ml h-1) whereas VAS values at rest were comparable (p < 0.1) at all measurepoints. Intensity of motor block and side effects did not differ between groups. The present study shows that continuous epidural infusion of 0.2% ropivacaine alone provides comparable analgesia at rest, but less profound pain relief during mobilisation within the first 24 hours after major abdominal surgery than bupivacaine 0.125% with sufentanil.


SaeR binds a consensus sequence within virulence gene promoters to advance USA300 pathogenesis.
This investigation examines the role of the SaeR/S 2-component system in USA300, a prominent circulating clone of community-associated methicillin-resistant Staphylococcus aureus. Using a saeR/S isogenic deletion mutant of USA300 (USA300DeltasaeR/S) in murine models of sepsis and soft-tissue infection revealed that this sensory system is critical to pathogenesis of USA300 during both superficial and invasive infection. Oligonucleotide microarray and real-time reverse-transcriptase polymerase chain reaction identified numerous extracellular virulence genes that are down-regulated in USA300DeltasaeR/S. Unexpectedly, an up-regulation of mecA and mecR1 corresponded to increased methicillin resistance in USA300DeltasaeR/S. 5'-RACE analysis defined transcript start sites for sbi, efb, mecA, lukS-PV, hlb, SAUSA300_1975, and hla, to underscore a conserved consensus sequence within promoter regions of genes under strong SaeR/S transcriptional regulation. Electrophoretic mobility shift assay experiments illustrated direct binding of SaeR(His) to promoter regions containing the conserved consensus sequence. Collectively, the findings of this investigation demonstrate that SaeR/S directly interacts with virulence gene promoters to significantly influence USA300 pathogenesis.


Neuroleptic malignant syndrome associated with risperidone and olanzapine in first-episode schizophrenia.
This case report describes an 18-year-old African American male who presented to a state psychiatric facility with symptoms of bizarre behavior, delusions and auditory hallucination. He was diagnosed with schizophrenia and treated with eight days of risperidone followed by five days of olanzapine. During this two-week period, this patient began to develop signs and symptoms of neuroleptic malignant syndrome (NMS). Two weeks after all neuroleptic medications were discontinued, all his vital signs and labs returned to baseline. This case report and others from literature suggest that risperidone and olanzapine may induce NMS in some patients.


Knee Implant Identification by Fine-Tuning Deep Learning Models.
Identification of implant model from primary knee arthroplasty in pre-op planning of revision surgery is a challenging task with added delay. The direct impact of this inability to identify the implants in time leads to the increase in complexity in surgery. Deep learning in the medical field for diagnosis has shown promising results in getting better with every iteration. This study aims to find an optimal solution for the problem of identification of make and model of knee arthroplasty prosthesis using automated deep learning models. Deep learning algorithms were used to classify knee arthroplasty implant models. The training, validation and test comprised of 1078 radiographs with a total of 6 knee arthroplasty implant models with anterior-posterior (AP) and lateral views. The performance of the model was calculated using accuracy, sensitivity, and area under the receiver-operating characteristic curve (AUC), which were compared against multiple models trained for comparative in-depth analysis with saliency maps for visualization. After training for a total of 30 epochs on all 6 models, the model performing the best obtained an accuracy of 96.38%, the sensitivity of 97.2% and AUC of 0.985 on an external testing dataset consisting of 162 radiographs. The best performing model correctly and uniquely identified the implants which could be visualized using saliency maps. Deep learning models can be used to differentiate between 6 knee arthroplasty implant models. Saliency maps give us a better understanding of which regions the model is focusing on while predicting the results.


Granulocyte-macrophage colony-stimulating factor enhances the efficacy of hepatitis B virus vaccine in previously unvaccinated haemodialysis patients.
The response to vaccination with recombinant hepatitis B virus (HBV) vaccine is poor in haemodialysis patients. A defect in the antigen-presenting cells may be responsible for this hyporesponsiveness. To overcome this and to improve the response to HBV vaccine in dialysis patients, we used granulocyte-macrophage colony-stimulating factor (GM-CSF) as a vaccine adjuvant. Fifteen consecutive patients with chronic renal failure (CRF), commenced on dialysis, were stratified to receive either 40microg HBV vaccine (Engerix-B) at 0, 1, 2 and 6 months (group A, n=9) or 3microg kg-1 GM-CSF (Leucomax) on day 1 followed by the vaccination schedule described above (group B, n=6). All patients were negative for hepatitis B surface antigen (HBsAg), antibodies to hepatitis C virus (anti-HCV) and human immunodeficiency virus (HIV) serology. Titres of antibody to HBsAg (HBsAb) were quantitatively assayed, using enzyme-linked immunosorbent assay (ELISA), at 1, 2, 6 and 7 months from the first dose of vaccination. Only 44% of the patients in group A developed protective antibody levels (mean HBsAb: 22 IU l-1) Fifty per cent of responders developed protective antibody levels (HBsAb >10 IU l-1) only after the fourth dose of vaccination. In contrast, all six patients (100%) in group B developed protective levels of HBsAb (mean HBsAb: 70 IU l-1) (P<0.02). Sixty-seven per cent of the responders were protected after only the second dose of vaccination (P=0.046). No serious adverse effects of GM-CSF were observed in group B. Hence, haemodialysis patients respond poorly to HBV vaccine. GM-CSF is a safe vaccine adjuvant capable of stimulating an earlier and a stronger antibody response to HBV vaccine in haemodialysis patients.


Statistical analysis of bicyclists' injury severity at unsignalized intersections.
This study investigated factors correlated with the severity of injuries sustained by bicyclists in bicycle-motor vehicle crashes at unsignalized intersections to develop site-specific countermeasures and interventions to improve bicycle safety. Crash data were extracted from accident reports entered into the Kentucky State Police's Kentucky Collision Database in 2002-2012. A partial proportional odds model was developed for data analysis. According to our modeling results, stop-controlled intersections, one-lane approaches, helmet usage, and lower speed limits were associated with decreased injury severity, whereas uncontrolled intersections, older (age > 55) drivers and bicyclists, child (age < 16) bicyclists, foggy and rainy weather, inadequate use of lights in dark conditions, and wet road surfaces were linked with increased injury severity. Based on these results, we suggest the development of educational programs focused on the following groups: child bicyclists, older bicyclists, and older drivers. Investigating and modifying street lighting could improve bicycle safety. Implementing road diets/traffic calming methods could create a safer traffic environment. Certain traffic control strategies (e.g., stop control) could be considered for uncontrolled intersections with high bicycle exposure, and helmet campaigns should be launched to increase helmet awareness and use. The study also suggests some interesting future research directions, including examining driver/bicyclist behaviors at uncontrolled intersections and studying the riding behaviors of child bicyclists in Kentucky.


Intracellular drug delivery nanocarriers of glutathione-responsive degradable block copolymers having pendant disulfide linkages.
Self-assembled micelles of amphiphilic block copolymers (ABPs) with stimuli-responsive degradation (SRD) properties have a great promise as nanotherapeutics exhibiting enhanced release of encapsulated therapeutics into targeted cells. Here, thiol-responsive degradable micelles based on a new ABP consisting of a pendant disulfide-labeled methacrylate polymer block (PHMssEt) and a hydrophilic poly(ethylene oxide) (PEO) block were investigated as effective intracellular nanocarriers of anticancer drugs. In response to glutathione (GSH) as a cellular trigger, the cleavage of pendant disulfide linkages in hydrophobic PHMssEt blocks of micellar cores caused the destabilization of self-assembled micelles due to change in hydrophobic/hydrophilic balance. Such GSH-triggered micellar destabilization changed their size distribution with an appearance of large aggregates and led to enhanced release of encapsulated anticancer drugs. Cell culture results from flow cytometry and confocal laser scanning microscopy for cellular uptake as well as cell viability measurements for high anticancer efficacy suggest that new GSH-responsive degradable PEO-b-PHMssEt micelles offer versatility in multifunctional drug delivery applications.


Enhanced electron transfer and hydrogen peroxide activation capacity with N, P-codoped carbon encapsulated CeO2 in heterogeneous electro-Fenton process.
Designing catalysts that can effectively activate oxygen and hydrogen peroxide is a huge challenge in electro-Fenton (EF) process. Considering the superior ability of electrons transport and activation of H2O2, ceria encapsulated with N, P-codoped carbon material was a promising catalyst for EF reaction. Herein, CeO2-NPCTX (where T and X represented the calcination temperature and the initial mass of CeO2, respectively) materials were synthesized via pyrolysis process and used as catalysts to degrade ciprofloxacin (CIP) in EF process. The results indicated that CeO2-NPC1000100 catalyst had good degradation performance under the optimal conditions. Compared with CeO2 and CeO2-NC1000100 catalysts, CeO2-NPC1000100 catalyst had more content of graphite N and more oxygen vacancies, which were beneficial to activation of oxygen and hydrogen peroxide. Scavenging experiments and electron paramagnetic resonance analysis confirmed ·O2- and ·OH were the main reactive oxygen species in the CIP degradation process. And three logical degradation routes of CIP were given. In addition, CeO2-NPC1000100 catalyst still had good stability after three times of continuous operation, and presented good universality for the treatment of a variety of antibiotic wastewaters. Finally, a convincing mechanism in the EF system with CeO2-NPC1000100 for CIP degradation was proposed.


Social interactions, information use, and the evolution of collective migration.
Migration of organisms (or cells) is typically an adaptive response to spatiotemporal variation in resources that requires individuals to detect and respond to long-range and noisy environmental gradients. Many organisms, from wildebeest to bacteria, migrate en masse in a process that can involve a vast number of individuals. Despite the ubiquity of collective migration, and the key function it plays in the ecology of many species, it is still unclear what role social interactions play in the evolution of migratory strategies. Here, we explore the evolution of migratory behavior using an individual-based spatially explicit model that incorporates the costs and benefits of obtaining directional cues from the environment and evolvable social interactions among migrating individuals. We demonstrate that collective migratory strategies evolve under a wide range of ecological scenarios, even when social encounters are rare. Although collective migration appears to be a shared navigational process, populations typically consist of small proportions of individuals actively acquiring directional information from their environment, whereas the majorities use a socially facilitated movement behavior. Because many migratory species face severe threat through anthropogenic influences, we also explore the microevolutionary response of migratory strategies to environmental pressures. We predict a gradual decline of migration due to increasing habitat destruction and argue that much greater restoration is required to recover lost behaviors (i.e., a strong hysteresis effect). Our results provide insights into both the proximate and ultimate factors that underlie evolved migratory behavior in nature.


A cellular model for sporadic ALS using patient-derived induced pluripotent stem cells.
Development of therapeutics for genetically complex neurodegenerative diseases such as sporadic amyotrophic lateral sclerosis (ALS) has largely been hampered by lack of relevant disease models. Reprogramming of sporadic ALS patients' fibroblasts into induced pluripotent stem cells (iPSC) and differentiation into affected neurons that show a disease phenotype could provide a cellular model for disease mechanism studies and drug discovery. Here we report the reprogramming to pluripotency of fibroblasts from a large cohort of healthy controls and ALS patients and their differentiation into motor neurons. We demonstrate that motor neurons derived from three sALS patients show de novo TDP-43 aggregation and that the aggregates recapitulate pathology in postmortem tissue from one of the same patients from which the iPSC were derived. We configured a high-content chemical screen using the TDP-43 aggregate endpoint both in lower motor neurons and upper motor neuron like cells and identified FDA-approved small molecule modulators including Digoxin demonstrating the feasibility of patient-derived iPSC-based disease modeling for drug screening.


Interactive patient-specific 3D approximation of scapula bone shape from 2D X-ray images using landmark-constrained statistical shape model fitting.
We report on an interactive tool for patientspecific 3D approximation of scapula bone shape from 2D X-ray images using landmark-constrained statistical shape model (SSM) fitting. The 3D localization of points on the 2D X-ray images was done through X-ray stereophotogrammetry. The inferior angle, acromion and the coracoid process were identified as reliable landmarks from the anteroposterior (AP) and oblique lateral views in a landmark selection study. The 3D scapula surface was approximated through fitting the scapula SSM to the 3D reconstructed coordinates of the selected landmarks. 3D point localization yielded average (X, Y, Z) coordinate reconstruction errors of (X=0.14, Y=0.07, Z=0.04) mm. The landmark-constrained fitting algorithm yielded an average error between the mean posterior model landmarks and the corresponding target landmarks of 0.49 mm using the three landmarks, and later 0.19 mm with sixteen landmarks. Average surface to surface error between the CT ground truth model and approximated model from within the dataset improved from 3.20 mm to 2.46 mm using three landmarks and using sixteen landmarks, respectively. Average surface to surface error between the CT ground truth model and the approximated model from outside the dataset improved from 4.28 mm to 3.20 mm using three landmarks and using sixteen landmarks, respectively.


Improving self-management skills: a whole systems approach.
This article outlines the rationale behind an approach developed at the National Primary Care Research and Development Centre (NPCRDC) to improve patients' abilities to self-manage their conditions with the support of services provided by the NHS. The approach is systematic and requires involvement of patients in the development of information, changing access arrangements to health services and promoting a patient-centred approach to care. A programme of research and development is currently being undertaken at The University of Manchester by researchers based at NPCRDC to investigate the effectiveness of this approach. The evidence base for the strategy under investigation is discussed and proposals are made for the role of nurses as facilitators for the self-management agenda in the NHS.


The role of psychotherapy in bipolar disorder.
Adjunctive psychosocial interventions for bipolar disorder target many of the issues that are not addressed by medication alone, including non-adherence, efficacy-effectiveness gap and functionality. Psychosocial interventions have been found to reduce relapse, particularly for the depressive pole, and improve functionality. Approaches such as psychoeducation, cognitive behaviour therapy, interpersonal and social rhythm therapy, and family therapy have shown benefits as adjunctive treatments. Each of the various psychosocial interventions has a unique emphasis, but they share common elements. These include: providing information and education; developing a personal understanding of the illness, such as triggers and early warning signs; having prepared strategies in place for early intervention, should symptoms of illness develop; and promoting a collaborative approach. Evidence to date supports the use of adjunctive psychosocial interventions in the management of bipolar disorder.


Intestinal TMEM16A control luminal chloride secretion in a NHERF1 dependent manner.
TMEM16A (Transmembrane protein 16A or Anoctamin1) is a calcium-activated chloride channel. (CaCC),that exerts critical roles in epithelial secretion. However, its localization, function, and regulation in intestinal chloride (Cl-) secretion remain obscure. Here, we show that TMEM16A protein abundance correlates with Cl- secretion in different regions of native intestine activated by the Ca2+-elevating muscarinic agonist carbachol (CCH). Basal, as well as both cAMP- and CCH-stimulated Isc, was largely reduced in Ano1 ± mouse intestine. We found CCH was not able to increase Isc in the presence of apical to serosal Cl- gradient, strongly supporting TMEM16A as primarily a luminal Cl- channel. Immunostaining demonstrated apical localization of TMEM16A where it colocalized with NHERF1 in mouse colonic tissue. Cellular depletion of NHERF1 in human colonic T84 cells caused a significant reduction of both cAMP- and CCH-stimulated Isc. Immunoprecipitation experiments revealed that NHERF1 forms a complex with TMEM16A through a PDZ-based interaction. We conclude that TMEM16A is a luminal Cl- channel in the intestine that functionally interacts with CFTR via PDZ-based interaction of NHERF1 for efficient and specific cholinergic stimulation of intestinal Cl- secretion.


Implementation of breast cancer screening.
Mammography, breast physical examination, and breast self-examination are recommended for breast cancer screening. Although in randomized trials of screening, mammography demonstrated a reduction in breast cancer mortality for women over 50, such data are not available for breast physical or self-examination. The Canadian National Breast Screening Study should provide data regarding the efficacy of breast physical examination alone. The World Health Organization has established trials to evaluate self-examination in the USSR and Germany. There is still controversy regarding routine mammographic screening and the optimal screening interval for women under age 50. The costs of mammographic screening remain a concern, but these costs can be lessened considerably through efficiency of mammographic services and a reduction in the number of false-positive interpretations that prompt biopsy. Primary care physicians can help to reduce breast cancer mortality by referring their patients for screening mammography and by performing breast physical examinations.


Impaired regeneration of dystrophin-deficient muscle fibers is caused by exhaustion of myogenic cells.
Duchenne muscular dystrophy is one of the most devastating myopathies. Muscle fibers undergo necrosis and lose their ability to regenerate, and this may be related to increased interstitial fibrosis or the exhaustion of satellite cells. In this study, we used mdx mice, an animal model of Duchenne muscular dystrophy, to assess whether muscle fibers lose their ability to regenerate after repeated cycles of degeneration-regeneration and to establish the role of interstitial fibrosis or exhaustion of satellite cells in this process. Repeated degenerative-regenerative cycles were induced by the injection of bupivacaine (33 mg/kg), a myotoxic agent. Bupivacaine was injected weekly into the right tibialis anterior muscle of male, 8-week-old mdx (N = 20) and C57Bl/10 (control, N = 10) mice for 20 and 50 weeks. Three weeks after the last injection, the mice were killed and the proportion of regenerated fibers was counted and reported as a fibrosis index. Twenty weekly bupivacaine injections did not change the ability of mdx muscle to regenerate. However, after 50 weekly bupivacaine injections, there was a significant decrease in the regenerative response. There was no correlation between the inability to regenerate and the increase in interstitial fibrosis. These results show that after prolonged repeated cycles of degeneration-regeneration, mdx muscle loses its ability to regenerate because of the exhaustion of satellite cells, rather than because of an increase in interstitial fibrosis. This finding may be relevant to cell and gene therapy in the treatment of Duchenne muscular dystrophy.


Implementation of virtual reality (VR) in diagnostics and therapy of nonaffective psychoses.
Immersive virtual reality is a technology that allows the user to immerse in the virtual world in isolation from external stimuli. It enables the simulation of different social situations, often impossible to arrange in reality, with high control over the confounding variables. Thanks to the VR realism, the viewer of this reality behaves similarly and experiences similar emotions to those in natural conditions, which results in high ecological validity of this environment, making it useful for diagnostics and therapy. This review, conducted in a narrative way, presents the results of observational and interventional research using immersive virtual reality (VR) in exploration of mechanisms generating psychotic symptoms (mainly in the scope of paranoia), as well as cognition and social functioning (research with the use of virtual avatars) in persons diagnosed with nonaffective psychosis. The research included in the review has been divided by the authors into two categories, depending on their type and the related level of reliability of the results. Moreover, the authors discuss technological aspects of VR, including the most important ways of presenting it, the differences between VR technology and classical neurocognitive tests, and the use of this technology for diagnostic purposes. As far as the treatment of psychotic disorders is concerned, the authors discuss VR interventions focused mainly on delusions and auditory hallucinations. Finally, the prospects for further development and use of VR technology in psychiatry are discussed.


Impact of Bridging Locoregional Therapies for Hepatocellular Carcinoma on Post-transplant Clinical Outcome.
Long waiting times due to ongoing organ shortage have led to increased utilization of locoregional therapies (LRTs) to bridge patients with hepatocellular carcinoma (HCC) to liver transplantation (LT). We performed this study to evaluate the impact of LRTs on post-LT outcomes. We conducted a retrospective study of patients who were transplanted for HCC at Stanford University Hospital between 2008 and 2018 (n = 302). We found that receipt of ≥5 LRTs was an independent and significant predictor of poor overall 5-year survival (58.3% vs. 83.3%; HR 2.26, p = .03), poor recurrence-free 5-year survival (51.9% vs. 80.4%; HR 2.12, p = .03), and was associated with higher rates of recurrence (25.0% vs. 7.4%, p = .001). Moreover, recurrent HCC was more likely to be the cause of death (58.3% vs. 41.7%, p = .04) in patients who received ≥5 LRTs. Also, patients who required ≥5 LRTs showed an overall lower rate of radiological complete response (46.9% vs. 97.8%, p = .001) and were more likely to have more advanced pathological stage tumors in the explant (65.6% vs. 29.6%, p < .001). In conclusion, receipt of ≥5 bridging LRTs prior to LT is associated with worse post-transplant clinical outcomes.


Preclinical efficacy of dual mTORC1/2 inhibitor AZD8055 in renal cell carcinoma harboring a TFE3 gene fusion.
Renal cell carcinomas (RCC) harboring a TFE3 gene fusion (TfRCC) represent an aggressive subset of kidney tumors. Key signaling pathways of TfRCC are unknown and preclinical in vivo data are lacking. We investigated Akt/mTOR pathway activation and the preclinical efficacy of dual mTORC1/2 versus selective mTORC1 inhibition in TfRCC. Levels of phosphorylated Akt/mTOR pathway proteins were compared by immunoblot in TfRCC and clear cell RCC (ccRCC) cell lines. Effects of the mTORC1 inhibitor, sirolimus, and the dual mTORC1/2 inhibitor, AZD8055, on Akt/mTOR activation, cell cycle progression, cell viability and cytotoxicity were compared in TfRCC cells. TfRCC xenograft tumor growth in mice was evaluated after 3-week treatment with oral AZD8055, intraperitoneal sirolimus and respective vehicle controls. The Akt/mTOR pathway was activated to a similar or greater degree in TfRCC than ccRCC cell lines and persisted partly during growth factor starvation, suggesting constitutive activation. Dual mTORC1/2 inhibition with AZD8055 potently inhibited TfRCC viability (IC50 = 20-50 nM) due at least in part to cell cycle arrest, while benign renal epithelial cells were relatively resistant (IC50 = 400 nM). Maximal viability reduction was greater with AZD8055 than sirolimus (80-90% versus 30-50%), as was the extent of Akt/mTOR pathway inhibition, based on significantly greater suppression of P-Akt (Ser473), P-4EBP1, P-mTOR and HIF1α. In mouse xenograft models, AZD8055 achieved significantly better tumor growth inhibition and prolonged mouse survival compared to sirolimus or vehicle controls. Akt/mTOR activation is common in TfRCC and a promising therapeutic target. Dual mTORC1/2 inhibition suppresses Akt/mTOR signaling more effectively than selective mTORC1 inhibition and demonstrates in vivo preclinical efficacy against TFE3-fusion renal cell carcinoma.


A scenario-based dieting self-efficacy scale: the DIET-SE.
The article discusses a scenario-based dieting self-efficacy scale, the DIET-SE, developed from dieter's inventory of eating temptations (DIET). The DIET-SE consists of items that describe scenarios of eating temptations for a range of dieting situations, including high-caloric food temptations. Four studies assessed the psychometric properties of the 11-item DIET-SE. Exploratory factor analysis (N = 392) and confirmatory factors analysis (N = 124) revealed three internally consistent and reliable factors representing challenges to adhere to a diet (high-caloric food temptations [HCF], social and internal factors [SIF], negative emotional events [NEE]). Convergent validity is established with other measures of dieting self-efficacy, as well as measures of eating disinhibition, susceptibility to hunger, and weight loss competency. Criterion-related validity is provided through the assessment of goal adherence, and predictive validity is established for dieters' actual food intake (N = 68). The DIET-SE represents a short, reliable, and valid scenario-based measure of dieting self-efficacy.


Efficacy of bacterial removal from instrumented root canals in vitro related to instrumentation technique and size.
This in vitro investigation assessed the efficacy of removing radioactively labeled bacteria from infected canals with 2 engine-driven rotary nickel titanium instrumentation techniques differing in sequence and apical enlargement size. A standard quantity of (3)H-thymidine-labeled Enterococcus faecalis (3.70 x 10(4) cpm, 2.0 x 10(7) colony-forming units) was used to inoculate the mesiobuccal canals of 50 extracted mandibular molars. The teeth were incubated for 5 days to allow infection of the surrounding dentin from the canals. Five of the teeth were used as controls to determine the number of cycles of irrigation and drying necessary to reduce the (3)H counts recovered from the canals to baseline levels. After this process, the unbound bacteria in the root canals of the remaining 45 teeth then were washed out with buffer until baseline levels of radioactivity were obtained. The mesiobuccal root of 1 of these 45 teeth was removed, decalcified, and digested, and the total radioactivity released from the root dentin was measured. Of the remaining 44 teeth, 22 then were instrumented with GT and Profile (Dentsply/Tulsa Dental Co, Tulsa, Okla) instruments to apical size #35 (group 1) and 22 teeth with Pow-R instruments (Moyco/Union Broach, York, Pa) to apical size #50 (group 2), in the presence of a standard quantity of phosphate-buffered saline solution placed in the canal. After instrumentation, the medium from each canal was collected with paper points and its radioactivity was counted with liquid scintillation spectrometry. The mean (3)H level recovered with instrumentation of canals in group 1 was 75 cpm (+/- 29, standard deviation) and in group 2 was 123 cpm (+/- 50, standard deviation). A 2-tailed Mann-Whitney test indicated that the radioactivity of samples from group 2 was significantly higher than that of samples from group 1. The results suggested that instrumentation to an apical size of #50, as performed with the Pow-R instruments, was more effective in debriding infected root canals than instrumentation to an apical size of #35, as performed with the GT and Profile instruments.


Changes in B lymphocyte function in rheumatoid arthritis and lupus nephritis after total lymphoid irradiation.
Patients with rheumatoid arthritis and patients with lupus nephritis underwent total lymphoid irradiation (TLI). Peripheral blood mononuclear cells from these patients showed a decrease in pokeweed mitogen-induced immunoglobulin secretion after TLI. A defect in non-T cells contributed to the decreased response. No defect in the response of non-T cells to T cell-independent activators of B cells, Staphylococcus aureus Cowan 1 and Epstein-Barr virus, was observed after TLI. We conclude that TLI induces a selective deficit in B cells, which respond to T cell-dependent mitogen stimulation.


Drug management of terminal symptoms in advanced cancer patients.
This review addresses recent research into drug management of terminal major symptoms such as breathlessness, terminal delirium and death rattle. High quality evidence on pharmacotherapy at the end of life is lacking. For delirium currently available data demonstrate no clear advantage of use of atypical antipsychotics. The roles of donepezil and modafinil in management of delirium are yet to be assessed. Regarding dyspnoea, one randomized trial demonstrated that midazolam, in combination treatment, may enhance the effect of morphine on terminal dyspnoea. A recent study suggested that glycopyrronium is at least as effective as hyoscine hydrobromide in dealing with death rattle. Finally, a randomized, placebo-controlled, double-blind trial evaluating dextromethorphan identified no significant improvement in pain management in combined treatment with morphine. Conducting well designed trials in the setting of terminal cancer is complex and ethically questionable. Most data retrieved are based mainly on clinical experience. Further high quality research is required to inform clinical recommendations regarding terminal care.


Rapid adjustment of guard-cell abscisic Acid levels to current leaf-water status.
Detached broad bean (Vicia faba L.) leaflets were water stressed; within 15 minutes, guard-cell abscisic acid (ABA) concentration increased ninefold. This result eliminates the apparent discrepancy raised by reports of no correlation between initial water-stress effects on stomata and leaf ABA concentration. Six hours after stress relief, guard-cell ABA concentration was near the prestress value, which would seem to implicate other factors in stress aftereffects on stomata.


Stoichiometry of proton translocation coupled to substrate oxidation in plant mitochondria.
The proton translocation coupled to the electron flux from succinate, exogenous NADH, and NAD(+)-linked substrates (malate and isocitrate) to cytochrome c and to oxygen was studied in purified potato (Solanum tuberosum) mitochondria using oxygen and ferricyanide pulse techniques. In the presence of valinomycin plus K(+) (used as a charge compensating cation), optimum values of H(+)/2 e(-) were obtained when low amounts of electron acceptors (oxygen or ferricyanide) were added to the mitochondria (1-2 nanogram [2 e(-)] equivalents per milligram protein). The stoichiometry of proton translocation to electron flux was unaffected in the presence of N-ethylmaleimide, an inhibitor of the Pi/H(+) symport. With succinate as substrate, H(+)/2 e(-) ratios were 4.0 +/- 0.2 and 3.7 +/- 0.3 with oxygen and ferricyanide as electron acceptors, respectively. With exogenous NADH, H(+)/2e(-) ratios were 4.1 +/- 0.9 and 3.4 +/- 0.2, respectively. The proton translocation coupled to the oxidation of NAD(+)-linked substrates (malate, isocitrate) was dependent upon the presence of adenylates (ADP, AMP, or ATP). For malate (+ glutamate) oxidation the observed H(+)/2 e(-) ratios were increased from 3.6 +/- 2.2 to 6.5 +/- 0.5 in the presence of 20 micromolar ADP.


The cytological mechanism of low fertility in the naked seed rice.
The low fertility of naked seed rice (NSR) was investigated by the following observations: somatic chromosome constitute, behavior of pollen mother cells (PMCs), the germination of mature pollen grains, the development of male and female gametes and the structure of the anther opening. The results indicated that somatic chromosomal number was 2n = 24, behavior of PMCs were normal and most of pollen grains could regularly develop further to mature male gametophytes in NSR. And dehiscence chamber and thickened endothecium cell (TEC) in numerous anthers of the NSR were developed abnormally after dicaryotic phase, result in few anthers complete opening and most partly opening or failure to opening, therefore much fewer of pollen grains attach on the stigma as compared with normal variety. Furthermore most of embryo sacs possessed abnormal structure and were sterile. All of above illustrated that the failure of the anther opening and the abortion of female gametophyte were main factors controlling the low seed-setting rate of the NSR.


Smoking and physical activity interrelations in health science students. Is smoking associated with physical inactivity in young adults?
Smoking and physical inactivity constitute a major public health concern for Greece. The purpose of this study was to examine smoking behaviour and physical activity (PA) in Greek health science students. A standardised questionnaire and the Greek version of IPAQ-short were given to 2000 health science students, randomly selected from five higher education institutions, in order to record smoking behaviour and PA status. All healthy young adults aged 19-30 years old were eligible. The final cohort size of the study was 1651 students (690 men). In the overall population, smoking prevalence was 37.6%, with 23.1% being heavy smokers (=21 cig/day). Smoking prevalence did not differ significantly between sexes, but heavy smoking was more prevalent in males. Age at smoking initiation was negatively associated with the daily number of cigarettes smoked (smoking volume), but only in females. The prevalence of health-enhancing PA (high PAclass) was only 14.2%, while 45.4% of the study population was classified as insufficiently active (low PAclass). Males were more physically active than females. Logistic regression analysis showed a strong inverse association between smoking and PA that was more pronounced in males. Smoking was associated with significantly decreased odds of being either moderately or highly physically active. Smoking volume was also negatively related with PA, but this relation was more pronounced in females. Smoking prevalence and rates of physical inactivity are considerably high in Greek health science students. Smoking was strongly and inversely associated with PA in this sample of Greek young adults.


Is immunoproliferative small intestinal disease uncommon in India?
Till date only three series of immunoproliferative small intestinal disease (IPSID) describing 22 patients have been reported from India. Seven patients with IPSID in two tertiary referral centers in India are included in the study. Diagnosis was based on typical clinical features [diarrhoea (7/7), weight loss (7/7), clubbing (6/7), fever (3/7), abdominal pain and lump (3/7)], biochemical evidence of malabsorption and duodenal biopsy findings. All patients were young males (mean age 29.8 +/- 11.8 years, range 17-53). Atypical features included gastric involvement (1/7), colonic involvement (1/7) and appearance of pigmented nails following anti-cancer chemotherapy (1/7) which disappeared six months after omitting doxorubin from chemotherapy regimen. Parasitic infestation was common. Ascaris lumbricoides (1/7), Giardia lamblia and hookworm (1/7), Strongyloides stercoralis and Trichuris trichura (1/7). In the latter patient S. stercoralis became disseminated after anti-malignant chemotherapy. One patient had gastric H. pylori infection. Four of the seven patients who were misdiagnosed as tropical sprue were treated with tetracycline. This raises doubt on efficacy of tetracycline alone in treatment of IPSID. One other patient was misdiagnosed and treated as intestinal tuberculosis. Early diagnosis and administration of chemotherapy may improve survival in this disease.


[A case of stage IV diffusely infiltrating colon cancer treated by surgical resection and chemoradiation therapy].
A 62-year-old male patient presented at the hospital because of left lower abdominal tumor. Based on preoperative examination and biopsy results, he was diagnosed with stage IV diffusely infiltrating colon cancer (scirrhous type) with paraaortic lymph node metastases. He underwent sigmoidectomy with D1 lymph node dissection and received systemic infusion of 5-FU 750 mg and l-LV 300 mg once a week. This chemotherapy produced no change in response in the paraaortic lymph node metastases for a long time. One year later, there were distant lymph node metastases including left inguinal and Virchow's lymph node, and systemic infusion of CPT-11 was performed. In addition, left inguinal lymph node was treated with irradiation therapy (total 50 Gy). The patient died of multiple organ failure 18 months after the operation. It is known that the prognosis in cases of diffusely infiltrating colorectal cancer is extremely poor. However, this case might suggest that intensive therapies with surgery and chemoradiation are useful in maintaining quality of life and improving survival.


[Pulmonary vasoreactivity testing in diagnosis and prognosis of pulmonary hypertension].
Right heart catheterization (RHC) is required for confirmation of the diagnosis of pulmonary hypertension (PH) and to asses the prognosis and etiology. Pulmonary vasoreactivity testing (PVT) is one of the most crucial parts of RHC especially in the suitable patient subsets of pulmonary arterial hypertension (PAH) defined by the most recent guidelines. PVT has substantial importance in defining the most appropriate treatment modality. The aim of this review is to provide a comprehensive review of the literature about PVT and the agents used in PVT, to emphasize the importance of this procedure when done with correct indications with a correct technique and to contribute in the accurate management of PH patients.


Epigenotype switching at the CD14 and CD209 genes during differentiation of human monocytes to dendritic cells.
Using an in vitro model for the differentiation of human blood-derived monocytes into dendritic cells (DCs), we have undertaken an investigation of epigenetic changes that take place at CD14 and CD209 (DC-SIGN) genes that encode cell surface antigens that are crucial for the function of monocytes and DCs, respectively. Upon differentiation the cell surface expression of CD14 is lost, whilst CD209 expression is increased. These reciprocal changes are associated with the loss of epigenetic markers of "activation" at the CD14 locus, but the acquisition of the same at the CD209 locus. There is little change in "repressive" histone marks and CpG methylation at the CD14 locus. By contrast there are changes in both the "repressive" histone marks and CpG methylation at the CD209 locus. In particular, two CpG dinucleotides, designated CpG2 and CpG3, show a marked demethylation at the CD209 promoter upon differentiation. These data provide insight into the epigenetic demands that are necessary to effect the reciprocal changes in expression of the CD14 and CD209 genes upon terminal differentiation of monocytes into DCs. For repression of the active CD14 gene the loss of "activation" histone modifications is likely necessary and sufficient for silencing. By contrast the activation of the silent CD209 gene appears to require an acquisition of "active" histone modifications and concomitant loss of both "repressive" histone marks and CpG methylation.


Closed-form Maker fringe formulas for poled polymer thin films in multilayer structures.
We report new closed-form expressions for Maker fringes of anisotropic and absorbing poled polymer thin films in multilayer structures that include back reflections of both fundamental and second-harmonic waves. The expressions, based on boundary conditions at each interface, can be applied to multilayer structures containing a buffer and a transparent conducting oxide layer, which might enhance multiple reflections of fundamental and second-harmonic waves inside a nonlinear thin film layer. This formulation facilitates Maker fringe analysis for a sample containing additional multilayer structures on either side of a poled polymer thin film. Experimental data and numerical simulations are given to indicate the importance of inclusion of such a reflective layer in analyses for reliable characterization of second-harmonic tensor elements.


Debra, a protein mediating lysosomal degradation, is required for long-term memory in Drosophila.
A central goal of neuroscience is to understand how neural circuits encode memory and guide behavior changes. Many of the molecular mechanisms underlying memory are conserved from flies to mammals, and Drosophila has been used extensively to study memory processes. To identify new genes involved in long-term memory, we screened Drosophila enhancer-trap P(Gal4) lines showing Gal4 expression in the mushroom bodies, a specialized brain structure involved in olfactory memory. This screening led to the isolation of a memory mutant that carries a P-element insertion in the debra locus. debra encodes a protein involved in the Hedgehog signaling pathway as a mediator of protein degradation by the lysosome. To study debra's role in memory, we achieved debra overexpression, as well as debra silencing mediated by RNA interference. Experiments conducted with a conditional driver that allowed us to specifically restrict transgene expression in the adult mushroom bodies led to a long-term memory defect. Several conclusions can be drawn from these results: i) debra levels must be precisely regulated to support normal long-term memory, ii) the role of debra in this process is physiological rather than developmental, and iii) debra is specifically required for long-term memory, as it is dispensable for earlier memory phases. Drosophila long-term memory is the only long-lasting memory phase whose formation requires de novo protein synthesis, a process underlying synaptic plasticity. It has been shown in several organisms that regulation of proteins at synapses occurs not only at translation level of but also via protein degradation, acting in remodeling synapses. Our work gives further support to a role of protein degradation in long-term memory, and suggests that the lysosome plays a role in this process.


The early magnetic resonance imaging features of the knee in juvenile idiopathic arthritis.
Early diagnosis of juvenile idiopathic arthritis (JIA) facilitates earlier more aggressive therapy, and improved outcome. Recognition of the features of early, untreated JIA on magnetic resonance imaging (MRI) will improve disease detection and expedite treatment. This study aims to highlight the relevant MRI features. MRI examinations of the knee joint were performed on 11 children with clinically confirmed, early, untreated JIA. The MRI images were obtained at a mean of 2 months after symptom onset and independently evaluated by two consultant paediatric radiologists. Abnormalities were found on all MRI examinations. Synovial hypertrophy, joint effusions, popliteal lymph nodes and soft tissue swelling were present in all patients. Gadolinium DTPA enhancement improved the detection of synovial hyperplasia. Metaphyseal splaying and condylar overgrowth were seen in five cases (41%), oedema of the lateral collateral ligament in two cases (18%) and superficial cartilage thinning in one case. Bony erosions and deep cartilage destruction were not demonstrated. MRI of the knee joint identifies early joint changes which are distinct from those in later disease. The presence of these features should alert the radiologist to the possible diagnosis of JIA and post gadolinium DTPA sequences should be performed. Gadolinium DPTA enhancement increases the sensitivity for the detection of inflammatory changes in JIA.


Dietetic internship programs in Canada: a comparison between 1935 and 1985.
Dietetic internships for most Canadian university graduates in 1935 were only available in the United States. The first priority of the student training committee established by CDA in 1936 was to integrate the isolated attempts at student training in Canada and standardize the training programs. Fifteen hospitals were tentatively approved by the committee and a standard course outline developed. Internship program development in 1985 continues to emphasize the establishment of national standards and the administration and accreditation of dietetic programs. Many changes have occurred in dietetic training programs between 1935 and 1985 to meet the increasing demands on our profession. The founding members of our Association recognized the need to establish and maintain quality dietetic practice in 1935. Their foresight, determination, and search for excellence in those early years ensured the future of the dietetic profession in Canada.


Prevalence and genetic characterization of Giardia and Cryptosporidium in cats from Italy.
One hundred and eighty one cats living in central Italy were tested for the presence of Giardia and Cryptosporidium infection by IFAT test and specific PCRs. Overall eight (4.4%) samples were IFAT-positive for Giardia. All the IFAT-positive samples for Giardia scored positive for the PCRs, and three more samples IFAT-negative generated PCR products leading to a total 6.1% molecular positivity rate for Giardia. All the examined samples were negative for Cryptosporidium. Sequencing of samples molecularly positive to Giardia indicated that three cats harbored the zoonotic Giardia duodenalis Assemblage A, whereas all other positive animals were infected with the feline-specific G. duodenalis Assemblage F. Phylogenetic analysis carried out on the sequences obtained supported the clustering of the isolates within Assemblages A and F. The results here presented provide data on the occurrence of Giardia genotypes in cats living in close contact with humans highlighting the potential importance of this protozoan disease for the public health.


Sex steroids modulate prolactin action in spontaneously luteinizing porcine granulosa cells in vitro.
Prolactin (20-1000 ng/ml) stimulated progesterone secretion in monolayer cultures of granulosa cells isolated from mature (greater than 8 mm) porcine follicles. The action of 17 beta-estradiol alone and the interaction between estradiol and prolactin in controlling steroidogenesis were critically dependent upon the duration of hormone treatment. Acute administration (within 36 h) of 17 beta-estradiol, but not 17 alpha-estradiol, suppressed both spontaneous and prolactin-stimulated progesterone production. Continuation of estrogen treatment beyond 48 h markedly increased progesterone secretion per cell and with prolactin produced synergistic enhancement of progesterone accumulation. However, 5 alpha-dihydrotestosterone also significantly augmented progesterone production after both acute and prolonged administration. The steroidogenic effect of prolactin was greater in the presence of 5 alpha-dihydrotestosterone, but did not elicit true synergism. Measurements of sex-steroids levels in porcine follicular fluids indicated that estrogen concentrations that were biologically active in vitro are similar to those attained in vivo. These observations suggest a major role for sex-steroid hormones both in the control of basal steroidogenesis and in the modulation of prolactin action in mature spontaneously luteinizing granulosa cells.


Declarative memory is critical for sustained advantageous complex decision-making.
Previous studies have reported conflicting evidence concerning the contribution of declarative memory to advantageous decision-making on the Iowa Gambling Task (IGT). One study, in which the measurement of psychophysiology during the task necessitated a 10-s delay between card selections, found that six participants with amnesia due to hippocampal damage failed to develop a preference for advantageous decks over disadvantageous decks [Gutbrod, K., Krouzel, C., Hofer, H., Muri, R., Perrig, W., & Ptak, R. (2006). Decision-making in amnesia: Do advantageous decisions require conscious knowledge of previous behavioural choices? Neuropsychologia, 44(8), 1315-1324]. However, a single-case study (where psychophysiology was not measured and no delay between card selections occurred) showed that an amnesic patient developed normal preference for advantageous decks [Turnbull, O. H., & Evans, C. E. (2006). Preserved complex emotion-based learning in amnesia. Neuropsychologia, 44(2), 300-306]. We sought to resolve these discrepant findings by examining IGT performances in five patients with profound amnesia (WMS-III General Memory Index M=63) and bilateral hippocampal damage caused by anoxia (n=4) or herpes simplex encephalitis (n=1). In one administration of the IGT, psychophysiology measurements were utilized and a 6-s delay was interposed between card selections. In a second administration, no delay between card selections was interposed. While age-, sex-, and education-matched healthy comparison participants showed significant learning with a gradual preference for advantageous decks in both conditions, amnesic patients, irrespective of IGT administration condition and extent of medial temporal lobe damage, failed to develop this preference. These findings strongly discount the possibility that the delay between card selections explains why amnesic participants fail to learn in the IGT, and suggest instead a significant role for medial temporal lobe declarative memory systems in the type of complex decision-making tapped by the IGT.


Advancing the adoption, integration and testing of technological advancements within existing care systems.
This manuscript reviews the work on uptake and dissemination of health information technologies in mental health populations and settings, with the goal of informing the future research agenda. We reviewed both the formal and "grey" literature describing the rates and correlates of uptake for electronic health records (EHRs) and personal health records (PHRs) for general and specialty mental health settings. Rates of uptake and use of EHRs and PHRs are low in general medical settings, and the limited evidence suggests even lower rates for specialty mental health settings. Many of the patient, provider and system-level characteristics associated with lower rates of use in general populations would be expected to be exacerbated in mental health settings. The findings suggest a need to better understand both the causes and strategies for overcoming barriers to uptake of health information technology (HIT) in mental health settings. Observational studies could help to better elucidate the barriers to adoption of HIT that are unique or disproportionate in mental health populations. Implementation science studies are needed to better identify strategies for addressing these barriers and optimizing uptake of mental health HIT interventions.


Depletion of Janus kinase-2 promotes neuronal differentiation of mouse embryonic stem cells.
Janus kinase 2 (JAK2), a non-receptor tyrosine kinase, is a critical component of cytokine and growth factor signaling pathways regulating hematopoietic cell proliferation. JAK2 mutations are associated with multiple myeloproliferative neoplasms. Although physiological and pathological functions of JAK2 in hematopoietic tissues are well-known, such functions of JAK2 in the nervous system are not well studied yet. The present study demonstrated that JAK2 could negatively regulate neuronal differentiation of mouse embryonic stem cells (ESCs). Depletion of JAK2 stimulated neuronal differentiation of mouse ESCs and activated glycogen synthase kinase 3ꞵ, Fyn, and cyclin-dependent kinase 5. Knockdown of JAK2 resulted in accumulation of GTPbound Rac1, a Rho GTPase implicated in the regulation of cytoskeletal dynamics. These findings suggest that JAK2 might negatively regulate neuronal differentiation by suppressing the GSK-3β/Fyn/CDK5 signaling pathway responsible for morphological maturation. [BMB Reports 2021; 54(12): 626-631].


[The "right choice" in medical imaging studies: the radiologist's viewpoint--application to lumbosciatic pain].
These past few years, very efficient imaging technologies, which are not traumatic for the patient, but often quite expensive, have been developed. Within the context of budgetary restrictions, these progress require a carefully planned strategy on the choice of investigations. First of all basic rules are defined: "the most simple act", "the direct way", "the direct exploitation of results", "the therapeutic relevance", "the multidisciplinary integration". Several erroneous types of behaviour are also described: "the most comprehensive set of examinations", "the examination in fashion", "the exaggerated medico-legal fear". Finally, these principles apply to a common clinical situation: lumbosciatic pain. Relevant examinations are first spotted: standard examination, myelography, discography, CT scan, MRI. A synthetic diagnostic approach is then suggested. The latter rules out any complementary study in the case of non-chronic common lombosciatic pain and in the absence of certain revealing signs. In the other cases, the correct combination of conventional radiology, nuclear medicine, CT and MRI is suggested, according to the clinical context.


The P-Rex1/Rac signaling pathway as a point of convergence for HER/ErbB receptor and GPCR responses.
Guanine nucleotide Exchange Factors (GEFs) are responsible for mediating GDP/GTP exchange for specific small G proteins, such as Rac. There has been substantial evidence for the involvement of Rac-GEFs in the control of cancer cell migration and metastatic progression. We have previously established that the Rac-GEF P-Rex1 is a mediator of actin cytoskeleton rearrangements and cell motility in breast cancer cells downstream of HER/ErbB receptors and the G-Protein Coupled Receptor (GPCR) CXCR4. P-Rex1 is highly expressed in luminal A and B breast cancer compared to normal mammary tissue, whereas expression is very low in basal breast cancer, and its expression correlates with the appearance of metastasis in patients. Here, we discuss the involvement of P-Rex1 as an effector of oncogenic/metastatic receptors in breast cancer and underscore its relevance in the convergence of receptor-triggered motile signals. In addition, we provide an overview of our recent findings describing a cross-talk between HER/ErbB receptors and CXCR4, and how this impacts on the activation of P-Rex1/Rac1 signaling, as well as highlight challenges that lie ahead. We propose a model in which P-Rex1 acts as a crucial node for the integration of upstream inputs from HER/ErbB receptors and CXCR4 in luminal breast cancer cells.


The phenotype of recovery VI: The association between life-history strategies, delay discounting, and maladaptive health and financial behaviors among individuals in recovery from alcohol use disorders.
The life-history theory is a well-established framework that predicts behaviors and explains how and why organisms allocate effort and resources to different life goals. Delay discounting (DD) is associated with risky behaviors and has been suggested as a candidate behavioral marker of addiction. Thus, we investigated the relationship between DD, life-history strategies, and engagement in risky behaviors among individuals in recovery from alcohol use disorder (AUD). Data from 110 individuals in recovery from addiction from The International Quit & Recovery Registry, an ongoing online registry designed to understand recovery phenotype, were included in the analysis. The association between life-history strategies, DD, engagement in risky behaviors, and remission status were assessed. Life-history strategy scores were significantly associated with DD rates and finance, health, and personal development behaviors after controlling for age, sex, race, ethnicity, years of education, marital status, smoking status, and history of other substance use. Remission status was significantly associated with life-history strategy, DD, drug use, fitness, health, and safe driving after controlling for age, sex, race, years of education, marital status, and smoking status. In addition, a mediation analysis using Hayes' methods revealed that the discounting rates partially mediated the association between remission status and life-history strategy scores. Life-history strategies and remission status are both significantly associated with DD and various health and finance behaviors among individuals in recovery from AUD. This finding supports the characterizations of DD as a candidate behavioral marker of addiction that could help differentiate subgroups needing special attention or specific interventions to improve the outcomes of their recovery. Future longitudinal studies are warranted to understand the relationships between changes in life-history strategies, DD, maladaptive health behaviors, and remission status over time.


Activation of toll-like receptor 4 (TLR4) by in vivo and in vitro exposure of rat epididymis to lipopolysaccharide from Escherichia Coli.
This study provides the first evidence that rat epididymis is fully capable of initiating an inflammatory response to lipopolysaccharide (LPS) from Escherichia coli through activation of Toll-like receptor 4 (TLR4). TLR4 functionality was demonstrated by in vivo LPS challenge, which induced a time- and dose-dependent activation of the transcription factor nuclear factor kappa B (NFKB) in caput and cauda epididymides. NFKB activation by LPS in caput epididymidis was abrogated when rats were pretreated with the NFKB inhibitor PDTC, confirming the specificity of this response. Within 2 h of LPS treatment (0.01 and 1 mg/kg, i.v.), NFKB activation in caput and cauda was accompanied by upregulation of Il1b, Nfkbia, and Cd14, but not Tlr4, mRNA. These effects, however, were not sustained after 24 h of LPS treatment. Lipopolysaccharide systemic effects were not restricted to epididymides, since Il1b, Nfkbia, and Cd14 mRNAs were also upregulated in other male reproductive tissues from LPS-treated rats (1 mg/kg, i.v., 2 h). Constitutive TLR4 was immunolocalized in some, but not all, epididymal epithelial cells and in interstitial cells, some of them identified as resident ED2-positive macrophages. No change in TLR4 immunostaining pattern was observed when epididymides from control and LPS-treated rats were compared (1 mg/kg, i.v., 2 h and 24 h). Significant NFKB activation was also achieved within 1 min of in vitro incubation of caput epididymidis with LPS (0.01-5 mug/ml), confirming that components for TLR4 signaling cascade activation are fully active in this tissue. This study contributes to a better understanding of the innate immune response in the epididymis and other tissues from the male reproductive tract.


Clinical and molecular characterization of a combined 17p13.3 microdeletion with partial monosomy 21q21.3 in a 26-year-old man.
We led a clinical and molecular characterization of a patient with mild mental delay and dysmorphic features initially referred for cytogenetic exploration of an azoospermia. We employed FISH and array CGH techniques for a better definition and refinement of a double chromosome aberration associating a 17p microdeletion with partial monosomy 21q due to 1:3 meiotic segregation of a maternal reciprocal translocation t(17;21)(p13.3;q21.2) revealed after banding analysis. Brain MRI depicted partial callosal and mild diffuse cerebral atrophies, but without expected signs of lissencephaly. The patient's karyotype formula was: 45,XY,der(17)t(17;21)(p13.3;q21.2)mat,-21. FISH study confirmed these rearrangements and array CGH analysis estimated the loss sizes to at least 635 kb on chromosome 17 and to 15.6 Mb on chromosome 21. The absence of lissencephaly and major brain malformations often associated with 17p terminal deletions could be attributed to the retention of PAFAH1B1, YWHAE and CRK genes. Dysmorphic features, moderate mental impairment and minor brain malformations could result from the 21q monosomy and particularly the partial deletion of the APP-SOD1 region. Azoospermia should result from gamete apoptosis induced by a control mechanism triggered in response to chromosome imbalances. Our study provides an additional case for better understanding and delineating both 17p and 21q deletions.


Diagnostic relevance of beta2-transferrin for the detection of cerebrospinal fluid fistulas.
The beta(2)-transferrin assay is a specific method to identify cerebrospinal fluid (CSF). Hitherto, this test has not been widely used for the routine screening of patients with suspected CSF leakage. The purpose of this study was to investigate the clinical relevance of the identification of beta(2)-transferrin by comparing the test results with other diagnostic measures and intraoperative findings. Case series. Retrospective analysis of 182 patients tested once or multiple times for beta(2)-transferrin. Information was obtained regarding different diagnostic procedures applied to diagnose CSF leakage. The effectiveness of those diagnostic measures was compared. The main indication to test for beta(2)-transferrin was posttraumatic rhinorrhea (25%), followed by spontaneous (22%) and postsurgical (22%) rhinorrhea. In 35 of 205 cases, beta(2)-transferrin was detected in the tested specimens. Thirteen of these required surgical intervention for treatment of the CSF fistula, and the leakage site was identified in all of them. Taking all results into consideration, the highest correlation was observed between the beta(2)-transferrin assay, intrathecal fluorescein application, and surgical exploration. The beta(2)-transferrin assay is a reliable method for confirming suspected CSF and should be used as a primary screening method in all patients with suspected CSF leakage. Although less invasive, the beta(2)-transferrin assay almost matches the high sensitivity achieved by exploratory surgery and intrathecal application of fluorescein. However, the possibility of bias should be carefully considered, and in particular, negative results should be critically compared with clinical symptoms and with results from other diagnostic procedures.


DFT study on magnetic interaction in an orbitally degenerate Ti3+ dimer complex.
The magnetic interaction in an orbitally degenerate transition metal dimer complex is investigated using a typical example of a d(1)-d(1) dimer complex, the Ti2Cl9(3-) cluster. The local orbital functions are defined by linear combinations of the molecular orbital functions which are calculated by density functional theory (DFT). In the DFT calculation, the Perdew-Burke-Ernzerhof (PBE) functional and hybrid PBE0 functional are utilized. The matrix elements of the effective Hamiltonian of the d-electrons are evaluated by the DFT calculation except for one parameter which is determined by comparing the zero-temperature magnetic susceptibility in the direction along the c axis χ(∥) with the experimental result. By the calculation with the PBE0 functional, the zero-temperature magnetic susceptibility in the perpendicular direction χ(⊥) and the temperature dependence of the susceptibilities in both directions agree with the experiment. On the other hand, by the calculation with the PBE functional, χ(⊥) is smaller than the experimental values because the on-site potential is underestimated.


Threading together patient expertise.
Patients are valuable sources of expertise for other patients in similar situations, but little is understood about the nature of this expertise. To address this knowledge gap, we investigated informational support as a mechanism for peers to help one another learn to cope with the breast cancer experience. We analyzed the types of problems discussed and recommendations offered by correspondents in three online breast cancer communities. Informational support was prevalent and directed towards problems in which correspondents were planning for future events or coping with emergent situations. Peers shared a wealth of patient expertise, including action strategies, recommended knowledge, suggested approaches, and information resources for dealing with problems. Our results highlight how peers are helping one another to learn. These findings bring insight to new support we could provide to patients for developing and sharing patient expertise, such as problem-based information organization and functionality for collaborative problem solving.


Selective Separation of Highly Similar Proteins on Ionic Liquid-Loaded Mesoporous TiO2.
Separating proteins from their mixtures is an important process in a great variety of applications, but it faces difficult challenges as soon as the proteins are simultaneously of similar sizes and carry comparable net charges. To develop both efficient and sustainable strategies for the selective separation of similar proteins and to understand the underlying molecular mechanisms to enable the separation are crucial. In this work, we propose a novel strategy where the cholinium-based amino acid [Cho][Pro] ionic liquid (IL) is used as the trace additive and loaded physically on a mesoporous TiO2 surface for separating two similar proteins (lysozyme and cytochrome c). The observed selective adsorption behavior is explained by the hydration properties of the [Cho][Pro] loaded on the TiO2 surface and their partially dissociated ions under different pH conditions. As the pH is increased from 5.0 to 9.8, the degree of hydration of IL ions also increases, gradually weakening the interaction strength of the proteins with the substrates, more for lysozymes, leading to their effective separation. These findings were further used to guide the detection of the retention behavior of a binary mixture of proteins in high-performance liquid chromatography, where the introduction of ILs did effectively separate the two similar proteins. Our results should further stimulate the use of ILs in the separation of proteins with a high degree of mutual similarity.