Disparate mechanisms of antifolate resistance provoked by methotrexate and its metabolite 7-hydroxymethotrexate in leukemia cells: implications for efficacy of methotrexate therapy.
Methotrexate (MTX) is one of the leading drugs in the treatment of leukemia, but extensive metabolism to 7-hydroxymethotrexate (7-OHMTX) can limit its therapeutic efficacy. In this study we investigated whether 7-OHMTX itself can provoke anti-folate resistance that may further disrupt MTX efficacy. For this purpose, we developed resistance to 7-OHMTX as well as MTX in 2 human leukemia cell lines (CCRF-CEM and MOLT-4) by stepwise exposure to increasing concentrations of 7-OHMTX and MTX. Consequently, both leukemia cell lines displayed marked levels of resistance to 7-OHMTX (> 10-fold) and MTX (> 75-fold). The underlying mechanism of resistance in the MTX-exposed cells was a marked decrease (> 10-fold) in reduced folate carrier (RFC)-mediated cellular uptake of MTX. This was associated with transcriptional silencing of the RFC gene in MTX-resistant CCRF-CEM cells. In contrast, the molecular basis for the resistance to 7-OHMTX was due solely to a marked decreased (> 95%) in folylpolyglutamate synthetase (FPGS) activity, which conferred more than 100-fold MTX resistance upon a short-term exposure to this drug. This is the first demonstration that 7-OHMTX can provoke distinct modalities of antifolate resistance compared with the parent drug MTX. The implications of this finding for MTX efficacy and strategies to circumvent MTX resistance are discussed.


Individual patient data meta-analysis of enoxaparin vs. unfractionated heparin for venous thromboembolism prevention in medical patients.
Unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) are both recommended for venous thromboembolism (VTE) prophylaxis in hospitalized medical patients. To perform an individual patient data meta-analysis to evaluate the relative efficacy and safety of the LMWH enoxaparin and UFH in preventing VTE in hospitalized medical patients. Randomized clinical trials comparing subcutaneous enoxaparin (4000 IU once-daily) and UFH (5000 IU subcutaneous two- or three-times daily) for VTE prevention were identified by a systematic search. Individual patient data were obtained from each eligible trial. Overall, four trials were eligible, including 3600 patients randomized to receive enoxaparin (n = 1799) or UFH (n = 1801). Median patient age was 71 years, and 49.3% were female. Compared with UFH, enoxaparin was associated with risk reductions of 37% for total VTE [relative risk (RR) 0.63, 95% confidence interval (CI) 0.51-0.77] and 62% for symptomatic VTE (RR 0.38, 95% CI 0.17-0.85) at day 15. RR for total VTE in stroke and non-stroke patients was 0.59 (95% CI 0.47-0.74) and 0.87 (95% CI 0.51-1.50), respectively. Major bleeding rates were consistently low and similar between treatment groups at day 15 (RR 1.13, 95% CI 0.53-2.44). There was a trend towards reduced risk for mortality in patients receiving enoxaparin (RR 0.83, 95% CI 0.64-1.08), compared with UFH. Enoxaparin significantly reduces VTE in hospitalized medical patients, compared with UFH, without increasing the risk for major bleeding, and was associated with a trend towards reduced all-cause mortality.


Cerebrospinal fluid biomarkers for Alzheimer's disease in Down syndrome.
Down syndrome (DS), present in nearly six million people, is associated with an extremely high risk to develop Alzheimer's disease (AD). Amyloid-β and tau pathology are omnipresent from age 40 years onward, but clinical symptoms do not appear in all DS individuals. Dementia diagnostics is complex in this population, illustrating the great need for predictive biomarkers. Although blood biomarkers have not yet proven useful, cerebrospinal fluid (CSF) biomarkers (low amyloid-β42, high t-tau, and high p-tau) effectively contribute to AD diagnoses in the general population and are increasingly used in clinical practice. Surprisingly, CSF biomarkers have been barely evaluated in DS. Breaking the taboo on CSF analyses would finally allow for the elucidation of its utility in (differential) diagnoses and staging of disease severity. A sensitive and specific biomarker profile for AD in DS would be of paramount importance to daily care, adaptive caregiving, and specific therapeutic interventions.


Dependency of intraocular pressure elevation and glaucomatous changes in DBA/2J and DBA/2J-Rj mice.
In this study parameters relevant for glaucoma in DBA/2J (D2J) mice were compared with those in age-matched DBA/2J-Rj (D2Rj) mice, to challenge the postulated role of D2J mice as a model for secondary high-tension glaucoma. Genotyping for three known short nucleotide polymorphisms (SNPs) in the Tyrp1 gene and the Gpnmb gene by MALDI-TOF-MS and immunohistochemical staining for Gpnmb was performed in D2J and D2Rj mice. Twelve C57Bl/6 (B6), 8 D2Rj, and 11 D2J mice between 1 and 4 months of age were screened qualitatively and quantitatively for morphologic differences within the anterior eye segment. The IOP progression of 25 D2Rj and 18 D2J mice were investigated between 4 to 10.5 months after birth. At the end of this study, in 10 randomly selected individuals of each D2J and D2Rj cohort, correlation of IOP progression and optic nerve damage were determined in each eye. D2J and D2Rj strains were homozygous for both Tyrp 1 amino acid substitutions, so far only described in D2J mice. The Gpnmb(R150X) point mutation present in D2J mice was not detected in D2Rj. Accordingly, immunoreactivity (IR) for Gpnmb was present only in D2Rj and B6 eyes, but not in D2J. Compared with B6, both DBA/2 mice (D2) showed a significantly narrowed chamber angle caused by an anteriorly displaced ciliary body. IOP measurements showed an average IOP of approximately 14 mm Hg between age 4 and 7 months in D2Rj, which decreased to approximately 11 mm Hg in the period from 8 to 10.5 months. In D2J the average IOP showed a steady increase in the observed period from 4 to 10.5 months (from 8.65 to 15.58 mm Hg). Individuals with IOP peaks up to 30 mm Hg were detected in D2Rj, but none of these mice showed signs of an optic neuropathy after 10.5 months. In contrast, 30% of the investigated D2J mice at the age of 10.5 months showed a severe optic neuropathy. Individual data analyses, however, showed no significant correlation between elevated IOP and glaucomatous changes within the D2J population. Individual correlations of IOP course with axon loss in the single eyes confirmed that in D2J mice, hypertension is not the only causative factor in glaucomatous optic neuropathy. For further investigations on the pathogenesis of glaucoma in D2J mice, the D2Rj strain without a Gpnmb(R150X) mutation and without glaucomatous changes, but with individual IOP elevation, can be used as an interstrain control for D2J.


Catechol O-methyltransferase haplotype predicts immediate musculoskeletal neck pain and psychological symptoms after motor vehicle collision.
Genetic variations in the catechol-O-methyltransferase (COMT) gene have been associated with experimental pain and risk of chronic pain development, but no studies have examined genetic predictors of neck pain intensity and other patient characteristics after motor vehicle collision (MVC). We evaluated the association between COMT genotype and acute neck pain intensity and other patient characteristics in 89 Caucasian individuals presenting to the emergency department (ED) after MVC. In the ED in the hours after MVC, individuals with a COMT pain vulnerable genotype were more likely to report moderate-to-severe musculoskeletal neck pain (76 versus 41%, RR = 2.11 (1.33-3.37)), moderate or severe headache (61 versus 33%, RR = 3.15 (1.05-9.42)), and moderate or severe dizziness (26 versus 12%, RR = 1.97 (1.19-3.21)). Individuals with a pain vulnerable genotype also experienced more dissociative symptoms in the ED, and estimated a longer time to physical recovery (median 14 versus 7 days, P = .002) and emotional recovery (median 8.5 versus 7 days, P = .038). These findings suggest that genetic variations affecting stress response system function influence the somatic and psychological response to MVC, and provide the first evidence of genetic risk for clinical symptoms after MVC. The association of COMT genotype with pain symptoms, psychological symptoms, and recovery beliefs exemplifies the pleiotropic effects of stress-related genes, which may provide the biological substrate for the biopsychosocial model of post-MVC pain. The identification of genes associated with post-MVC symptoms may also provide new insights into pathophysiology.


Robust Interfacial Effect in Multi-interface Environment through Hybrid Reconstruction Chemistry for Enhanced Energy Storage.
Electrochemical-oxidation-driven reconstruction has emerged as an efficient approach for developing advanced materials, but the reconstructed microstructure still faces challenges including inferior conductivity, unsatisfying intrinsic activity, and active-species dissolution. Herein, we present hybrid reconstruction chemistry that synergistically couples electrochemical oxidation with electrochemical polymerization (EOEP) to overcome these constraints. During the EOEP process, the metal hydroxides undergo rapid reconstruction and dynamically couple with polypyrrole (PPy), resulting in an interface-enriched microenvironment. We observe that the interaction between PPy and the reconstructed metal center (i.e., Mn > Ni, Co) is strongly correlated. Theoretical calculation results demonstrate that the strong interaction between Mn sites and PPy breaks the intrinsic limitation of MnO2, rendering MnO2 with a metallic property for fast charge transfer and enhancing the ion-adsorption dynamics. Operando Raman measurement confirms the promise of EOEP-treated Mn(OH)2 (E-MO/PPy) to stably work under a 1.2 V potential window. The tailored E-MO/PPy exhibits a high capacitance of 296 F g-1 at a large current density of 100 A g-1. Our strategy presents breakthroughs in upgrading the electrochemical reconstruction technique, which enables both activity and kinetics engineering of electrode materials for better performance in energy-related fields.


Measurement of external shame: an inside view.
The aims of this study were to investigate the construct validity of the Other as Shamer scale (OAS) using confirmatory factor analysis (CFA) and to examine the psychometric properties of its Italian version in a sample of 687 nonclinical individuals. The CFA results indicated that the hypothesized hierarchical model (with 1 higher order factor and 3 first-order factors) was the best fitting solution. Cronbach's alpha indexes, as well as test-retest stability, provided satisfactory results. Correlations of the OAS total score and its subscales with the Beck Depression Inventory-II (rs = .30-.48) and the Teate Depression Inventory (rs = .32-.45) were both substantial and significant (p < .01). Receiver operating characteristic curves were constructed to indicate sensitivity and specificity of the OAS and its subscales when determining those nonclinical subjects who met clinical thresholds for depression symptoms. A series of cutoff scores for the OAS scale and its subscales was developed, with sensitivity values between .70 and .62, and specificity values between .71 and .62, indicating good to fair discrimination between the 2 groups (depressed vs. nondepressed). The theoretical and practical implications of these results were discussed.


The role of information system in multiple sclerosis management.
Multiple sclerosis (MS) is a chronic disease of central nervous system. The multiple sclerosis information system (MSIS), such as other information system (IS), depends on identification, collection and processing of data for producing useful information. Lack of the integrated IS for collecting standard data causes undesirable effects on exchanging, comparing, and managing. The aim of this study was to recognize the role of the IS in the MS management and determine the advantages and barriers in implementing of the MSIS. The present study was a nonsystematized review that was done in order to recognize the role of the IS in the MS management. In this study, electronic scientific resources such as scientific magazines and books and published topics at conferences were used. We used key words (IS, chronic disease management, and multiple sclerosis), their combination or their synonyms in title, key words, abstracts, and text of English articles and published reports from 1980 until 2013, and by using search engines such as Google, Google Scholar and scientific databases and electronic issues such as iPubMed, sufficiently important difference, Scopus, Medlib, and Magiran for gathering information. More than 200 articles and reports were collected and assessed and 139 of them. Findings showed that the MSIS can reduce of disease expenses through continuously collecting correct, accurate, sufficient, and timely patients and disease nature information; recoding; editing; processing; exchanging, and distributing among different health care centers. Although the MSIS has many advantages; but, we cannot ignore cultural, economic, technical, organizational, and managerial barriers. Therefore, it is necessary to do studies for preventing, reducing, and controlling them. One of the ways is to recognize the advantages of the MSIS and usage information technology in optimizing disease management.


Comparison of bone SUV obtained from different SPECT/CT systems.
In bone scintigraphy, it is difficult to compare quantitative values, such as standardized uptake value (SUV), obtained from 2 different single-photon emission computed tomography combined with computed tomography (SPECT/CT) devices owing to differences of imaging acquisition and analysis methods. Therefore, the purpose of this study was to compare the SUV obtained from different SPECT/CT devices using the ratio to normal bone, and to analyze the correlation between them. A total of 27 prostate cancer patients who underwent bone scintigraphy either using Symbia T16 or Symbia Intevo (Siemens Medical Solutions, Erlangen, Germany) were retrospectively analyzed. In each patient, spherical voxels of interest were placed on the following 5 reference points: humeral head (humerus), femoral neck (femur), lower part of the ilium (ilium), first lumbar vertebra (L1), fifth lumbar vertebra (L5), and the maximum SUV (SUVmax) and average SUV (SUVave) of these regions were measured. The mean and variance of SUVave (humerus) was the smallest on both SPECT/CT. To compare the SUV obtained from the 2 devices, the SUVave ratio and SUVmax ratio of each region were calculated as the SUVave and SUVmax of each region divided by the SUVave of the humeral head in each patient. Median values of the SUVmax ratio and SUVave ratio of each region showed similar tendencies in both devices, with correlation coefficients between 0.93 and 1.19. Our results demonstrated that by expressing the quantitative value of SUVave of each region as a ratio to the SUVave of the humeral head, accumulation in the targeted bone can be compared even when the imaging acquisition and analysis methodsare different.


Fluoxetine Affects Differentiation of Midbrain Dopaminergic Neurons In Vitro.
Recent meta-analyses found an association between prenatal exposure to the antidepressant fluoxetine (FLX) and an increased risk of autism in children. This developmental disorder has been related to dysfunctions in the brains' rewards circuitry, which, in turn, has been linked to dysfunctions in dopaminergic (DA) signaling. The present study investigated if FLX affects processes involved in dopaminergic neuronal differentiation. Mouse neuronal precursors were differentiated into midbrain dopaminergic precursor cells (mDPCs) and concomitantly exposed to clinically relevant doses of FLX. Subsequently, dopaminergic precursors were evaluated for expression of differentiation and stemness markers using quantitative polymerase chain reaction. FLX treatment led to increases in early regional specification markers orthodenticle homeobox 2 (Otx2) and homeobox engrailed-1 and -2 (En1 and En2). On the other hand, two transcription factors essential for midbrain dopaminergic (mDA) neurogenesis, LIM homeobox transcription factor 1 α (Lmx1a) and paired-like homeodomain transcription factor 3 (Pitx3) were downregulated by FLX treatment. The stemness marker nestin (Nes) was increased, whereas the neuronal differentiation marker β3-tubulin (Tubb3) decreased. Additionally, we observed that FLX modulates the expression of several genes associated with autism spectrum disorder and downregulates the estrogen receptors (ERs) α and β Further investigations using ERβ knockout (BERKO) mDPCs showed that FLX had no or even opposite effects on several of the genes analyzed. These findings suggest that FLX affects differentiation of the dopaminergic system by increasing production of dopaminergic precursors, yet decreasing their maturation, partly via interference with the estrogen system.


Outcome of rest with or without bandaging for treatment of carpal flexural contracture deformities in puppies: 47 puppies and 75 joints (2000-2018).
To review outcome of dogs with carpal flexural contracture deformities treated with rest alone or with rest and bandaging. 47 dogs (75 joints). Medical records of dogs with unilateral or bilateral carpal flexural contracture deformities were reviewed, and dogs were grouped according to deformity severity grade (graded on a scale from 1 to 3) at the time of diagnosis. Two treatment groups were compared: rest only and rest with a modified Robert-Jones bandage. All dogs were reevaluated weekly until recovery (ie, resolution of the deformity and lameness). All dogs responded to conservative management, with all dogs regaining full extension of the antebrachiocarpal joint and ambulating normally at the time of the final visit. Mean ± SD time from initial diagnosis to recovery (ie, resolution of the deformity and lameness) was 2.9 ± 2.2 weeks (median, 2 weeks; range, 1 to 9 weeks). For dogs with grade 1 or 2 severity, mean time to recovery did not differ significantly between treatment groups. For dogs with grade 3 severity, however, mean time to recovery was significantly shorter for dogs treated with rest and bandaging than for dogs treated with rest alone. Results suggested that conservative management (rest alone or rest and bandaging) was a successful treatment option for puppies with carpal flexural contracture deformity and that bandaging resulted in a shorter time to recovery for dogs that were severely affected.


The first genome sequence of Anopheles squamous from Macha, Zambia.
Despite efforts to minimize the impacts of malaria and reduce the number of primary vectors, malaria has yet to be eliminated in Zambia. Understudied vector species may perpetuate malaria transmission in pre-elimination settings. Anopheles squamosus is one of the most abundantly caught mosquito species in southern Zambia and has previously been found with Plasmodium falciparum sporozoites, a causal agent of human malaria. This species may be a critical vector of malaria transmission, however, there is a lack of genetic information available for An. squamosus. We report the first genome data and the first complete mitogenome (Mt) sequence of An. squamosus. The sequence was extracted from one individual mosquito from the Chidakwa area in Macha, Zambia. The raw reads were obtained using Illumina Novaseq 6000 and assembled through NOVOplasty alignment with related species. The length of the An. squamosus Mt was 15,351 bp, with 77.9 % AT content. The closest match to the whole mitochondrial genome in the phylogenetic tree is the African malaria mosquito, Anopheles gambiae. Its genome data is available through National Center for Biotechnology Information (NCBI) Sequencing Reads Archive (SRA) with accession number SRR22114392. The mitochondrial genome was deposited in NCBI GenBank with the accession number OP776919. The ITS2 containing contig sequence was deposited in GenBank with the accession number OQ241725. Mitogenome annotation and a phylogenetic tree with related Anopheles mosquito species are provided.


Effects of IL-3 on promoter usage, attenuation and antisense transcription of the c-myc oncogene in the IL-3-dependent Ba/F3 early pre-B cell line.
The c-myc gene uses two major promoters, P1 and P2, for production of mRNA. In most proliferating normal cells, transcripts initiated from P2 predominate over the ones from P1. Furthermore, transcription of normal and translocated c-myc genes is bidirectional and overlapping. In this study, we have measured the effects of interleukin 3 (IL-3) deprivation and restimulation on c-myc promoter usage in the IL-3-dependent pre-B cell line Ba/F3. The rapid drop in c-myc mRNA expression observed in Ba/F3 cells upon IL-3 deprivation is reversible upon restimulation with interleukin. The use of P1 and P2 promoters, as judged by P1/P2 ratio, shifted from 0.2 in cells in exponential growth or in quiescence to 1.3 following IL-3 stimulation. This change was not due to selective instability of one of the two myc transcripts. In vitro nuclear run-on experiments indicated that IL-3 addition resulted in a large release of transcriptional attenuation, as well as a significant increase in transcriptional initiation. These results are consistent with the hypothesis that P1/P2 promoter usage is involved in the control of transcriptional elongation. Furthermore, deprivation of IL-3 resulted in a dramatic increase in antisense transcription, whereas little change was observed in the rate of initiation of elongation of c-myc mRNA precursors. This correlation suggests a negative role for antisense transcription in expression of c-myc mRNA levels in IL-3-deprived Ba/F3 cells.


Biomechanical performance of cranial implants with different thicknesses and material properties: A finite element study.
This study investigated the effect of implant thickness and material on deformation and stress distribution within different components of cranial implant assemblies. Using the finite element method, two cranial implants, differing in size and shape, and thicknesses (1, 2, 3 and 4 mm, respectively), were simulated under three loading scenarios. The implant assembly model included the detailed geometries of the mini-plates and micro-screws and was simulated using a sub-modeling approach. Statistical assessments based on the Design of Experiment methodology and on multiple regression analysis revealed that peak stresses in the components are influenced primarily by implant thickness, while the effect of implant material is secondary. On the contrary, the implant deflection is influenced predominantly by implant material followed by implant thickness. The highest values of deformation under a 50 N load were observed in the thinnest (1 mm) Polymethyl Methacrylate implant (Small defect: 0.296 mm; Large defect: 0.390 mm). The thinnest Polymethyl Methacrylate and Polyether Ether Ketone implants also generated stresses in the implants that can potentially breach the materials' yield limit. In terms of stress distribution, the change of implant thickness had a more significant impact on the implant performance than the change of Young's modulus of the implant material. The results indicated that the stresses are concentrated in the locations of fixation; therefore, the detailed models of mini-plates and micro-screws implemented in the finite element simulation provided a better insight into the mechanical performance of the implant-skull system.


Inhibition of nitric oxide synthetase at mid-gestation in rats is associated with increases in arterial pressure, serum tumor necrosis factor-alpha, and placental apoptosis.
Reduced uteroplacental perfusion and maternal cardiovascular dysfunction have been considered to be the main pathophysiological features of preeclampsia. In order to determine whether inhibition of nitric oxide synthetase (NOS) during the initial stage of placentation is associated with impaired placental development and maternal cardiovascular dysfunction, we studied the effect of N-nitro-arginine methyl ester (L-NAME), a NOS inhibitor, on morphological changes in the placenta, maternal blood pressure, and serum tumor necrosis factor-alpha (TNF-alpha) in pregnant rats during the initial stage of placentation. Pregnant Wister rats were treated during mid-gestation (days 8-14) with either L-NAME or saline. On day 20 of pregnancy the rats were killed, and maternal blood and placentas were extracted and examined. In comparison with pregnant saline-treated control rats (blood pressure 119 +/- 9 mm Hg), pregnant rats treated with L-NAME displayed significant hypertension (blood pressure 178 +/- 7 mm Hg), which continued even after the withdrawal of L-NAME administration (P < 0.01). In L-NAME-treated pregnant rats, morphological examination showed decreased populations of placental trophoblast lineages, and a significant increase in placental trophoblast apoptosis. Serum TNF-alpha levels at day 20 of pregnancy were significantly higher in treated pregnant rats (21.2 +/- 9.6 pg/ml) than in control pregnant rats (3.3 +/- 2.8 pg/ml) (P < 0.01). Inhibition of NOS at mid-gestation in pregnant rats is associated with increases in arterial pressure, placental apoptosis, and serum TNF-alpha, all of which have been implicated as being pathophysiological features of preeclampsia.


Do trauma systems work? A comparison of major trauma outcomes between Aberdeen Royal Infirmary and Massachusetts General Hospital.
Trauma is an important matter of public health and a major cause of mortality. Since the late 1980s trauma care provision in the United Kingdom is lacking when compared to the USA. This has been attributed to a lack of organisation of trauma care leading to the formation of trauma networks and Major Trauma Centres in England and Wales. The need for similar centres in Scotland is argued currently. We assessed the activity of two quite different trauma systems by obtaining access to comparative data from two hospitals, one in the USA and the other in Scotland. Aggregate data on 5604 patients at Aberdeen Royal Infirmary (ARI) from 1993 to 2002 was obtained from the Scottish Trauma Audit Group. A comparable data set of 16,178 patients from Massachusetts General Hospital (MGH). Direct comparison of patient demographics; injury type, mechanism and Injury Severity Score (ISS); mode of arrival; length of stay and mortality were made. Statistical analysis was carried out using Chi-squared and Cochran-Mantel-Haenszel. There were significant differences in the data sets. There was a higher proportion of penetrating injuries at MGH, (8.6% vs 2.6%) and more severely injured patients at MGH, patients with an ISS>16 accounted for nearly 22.1% of MGH patients compared to 14.0% at ARI. ISS 8-15 made up 54.6% of ARI trauma with 29.6% at MGH. Falls accounted for 50.1% at ARI and 37.9% at MGH. Despite the higher proportion of severe injuries at MGH and crude mortality rates showing no difference (4.9% ARI vs 5.2% MGH), pooled odds ratio of mortality was 1.4 (95% confidence interval 1.2-1.6) showing worse mortality outcomes at ARI compared to MGH. In conclusion, there were some differences in case mix between both data sets making direct comparison of the outcomes difficult, but the effect of consolidating major trauma on the proportion and number of severely injured patients treated in the American Level 1 centre was clear with a significant improvement in mortality in all injury severity groups.


The potential for antitick plants as components of an integrated tick control strategy.
Extracts from various African plants have been shown to possess strong acaricidal and/or tick repellent properties. Some of the plant extracts are capable of reducing tick feeding, molting, fecundity, and viability of eggs. Pasture grasses capable of repelling, trapping, and killing ticks have also been reported. These grasses possess hairs (trichomes) that retard ticks from climbing to the top of the grasses in order to attach themselves to passing animals. Furthermore, some grass species, such as Stylosanthes, produce viscous fluids that poison and kill ticks. Results obtained with antitick plants are presented in this paper and feasibility of using them in an integrated tick control strategy is discussed.


Ankle-Brachial Index as a Predictor of Mortality in Hemodialysis: A 5-Year Cohort Study.
Abnormal ankle-brachial index (ABI) has been found to be a strong predictor of mortality in some hemodialysis populations in studies with relatively short periods of follow-up, lower than 2 years. This study aimed to assess the predictive value of abnormal ABI as a risk factor for death among patients on maintenance hemodialysis after a 5-year follow-up. A total of 478 patients on hemodialysis for at least 12 months were included in the study. ABI measurement was performed using a mercury column sphygmomanometer and portable Doppler. Patients were divided into 3 groups according to ABI (low: <0.9; normal: 0.9 to 1.3; and high: >1.3) and followed for a 60-month period. The prevalence rates of low, normal and high ABI were 26.8%, 64.6% and 8.6%, respectively. The 5-year survival rate was lower in the groups with low ABI (44.1%, P<0.0001) and high ABI (60.8%, P= 0.025) than in the group with normal ABI (71.7%). Cox regression was used to evaluate the association between ABI and mortality, adjusting for potential confounders. Using normal ABI as reference, a low, but not a high ABI was found to be an independent risk factor for all-cause mortality (HR2.57; 95% CI, 1.84-3.57 and HR 1.62; 95% CI, 0.93-2.83, respectively). long-term survival rates of patients with either low or high ABI were lower than the one from those with normal ABI. However, after adjustment for potential confounders, only low ABI persisted as an independent risk factor for all-cause mortality among hemodialysis patients.


Duplication and diversification of insulin genes in ray-finned fish.
Insulin is a key hormone for the regulation of metabolism in vertebrates. Insulin is produced by pancreatic islet cells in response to elevated glucose levels and leads to the uptake of glucose by tissues such as liver and adipose tissue to store energy. Insulin also has additional functions in regulating development. Previous work has shown that the proglucagon gene, which encodes hormones counter regulating insulin, is duplicated in teleost fish, and that the peptide hormones encoded by these genes have diversified in function. I sought to determine whether similar processes have occurred to insulin genes in these species. Searches of fish genomes revealed an unexpected diversity of insulin genes. A triplication of the insulin gene occurred at the origin of teleost fish, however one of these three genes, insc, has been lost in most teleost fish lineages. The two other insulin genes, insa and insb, have been retained but show differing levels of selective constraint suggesting that they might have diversified in function. Intriguingly, a duplicate copy of the insa gene, which I named insab, is found in many fish. The coding sequence encoded by insab genes is under weak selective constraint, with its predicted protein sequences losing their potential to be processed into a two-peptide hormone. However, these sequences have retained perfectly conserved cystine residues, suggesting that they maintain insulin's three-dimensional structure and therefore might modulate the processing and secretion of insulin produced by the other genes.


From beads on a string to the pearls of regulation: the structure and dynamics of chromatin.
The assembly of eukaryotic chromatin, and the bearing of its structural organization on the regulation of gene expression, were the central topics of a recent conference organized jointly by the Biochemical Society and Wellcome Trust. A range of talks and poster presentations covered topical aspects of this research field and illuminated recent advances in our understanding of the structure and function of chromatin. The two-day meeting had stimulating presentations complemented with lively discourse and interactions of participants. In the present paper, we summarize the topics presented at the meeting, in particular highlighting subjects that are reviewed in more detail within this issue of Biochemical Society Transactions. The reports bring to life the truly fascinating molecular and structural biology of chromatin.


Determinants of urinary bisphenol A concentrations in Mexican/Mexican--American pregnant women.
Prenatal exposure to bisphenol A (BPA) may be associated with adverse health effects in the developing fetus; however, little is known about predictors of BPA exposure during pregnancy. We examined BPA exposure in 491 pregnant women from the Center for the Health Assessment of Mothers and Children of Salinas (CHAMACOS) cohort and explored the role of living in the United States on significant dietary predictors of BPA exposure. Women provided urine samples up to two times during pregnancy (n=866 total samples). We computed the intraclass correlation coefficient (ICC) to evaluate variability in concentrations between collections and used generalized estimating equation (GEE) models to assess predictors of exposure. Geometric mean (GSD) BPA concentrations were 0.9 (2.8)μg/L and 1.0 (2.6)μg/L at the first and second prenatal visits, respectively. We observed greater within- than between-woman variability in urinary BPA concentrations (ICC=0.22). GEE models suggest that women who lived in the United States their entire life had 38% (CI: -0.1, 89.3) higher urinary BPA concentrations compared with other immigrant women. Additionally, women who consumed ≥3 sodas per day or hamburgers three times a week or more had 58% (CI: 18.0, 112.1) and 20% (CI: -0.2, 45.2) higher urinary BPA concentrations, respectively, compared with women who consumed no sodas or hamburgers. A higher percentage of women who lived their entire life in the United States reported increased consumption of sodas and hamburgers compared with other immigrant women. Independent of other factors, BPA urinary concentrations were slightly higher when the sample was collected later in the day. As in previous studies, high within-woman variability in urinary BPA concentrations confirms that several samples are needed to properly characterize exposure during pregnancy. Results also suggest that some factors could be modified to minimize exposures during pregnancy in our study participants (e.g., reducing soda and hamburger intake) and that factors associated with acculturation might increase BPA concentrations.


Posttraumatic stress responses in children: awareness and practice among a sample of pediatric emergency care providers.
Research suggests that up to 4 of 5 children experience symptoms of an acute stress response (ASR) after a motor vehicle-related injury, and approximately 25% will develop posttraumatic stress disorder (PTSD). The degree to which physicians recognize this problem has not been reported. Our objective was to evaluate current awareness and practices of a cohort of pediatric emergency care providers regarding posttraumatic stress in children. Participants were identified from a list of the American Academy of Pediatrics Section on Emergency Medicine and surveyed on their awareness of ASR after motor vehicle-related injury, risk factors for developing PTSD, and practices regarding emergency department (ED) interventions. Surveys from physicians not practicing clinical emergency medicine were excluded. Of 322 surveys returned, 287 responses met inclusion criteria. Among these respondents, 198 (69%) were pediatric emergency medicine board certified or eligible and 260 (91%) practiced in a designated pediatric ED. Only 20 of 287 respondents (7%) believed that children were likely to develop symptoms of posttraumatic stress at levels previously described. Also in contrast to recent literature, 248 respondents (86%) felt that severity of injury was associated with future development of PTSD. Associated parental injury was identified accurately as a risk factor by 250 respondents (87%). Of interest, only 31 respondents (11%) were aware of any available tools to assess risk for PTSD. In addition, 56 of 287 respondents (20%) indicated that they would not use such tools in the ED, most commonly citing time and cost constraints. Finally, only 52 respondents (18%) reported giving any verbal guidance and only 9 (3%) provided any written instructions about posttraumatic stress to their patients and families. Findings suggest that physicians underestimate the likely development of an ASR and PTSD in the pediatric population. At present, few physicians offer written or even verbal instruction related to the development of posttraumatic symptoms. Physician education along with a systematic approach of assessment and intervention is necessary to address the gap between underrecognition of this concern and desired clinical practice.


Coordinated involvement of mast cells and T cells in allergic mucosal inflammation: critical role of the CC chemokine ligand 1:CCR8 axis.
CCL1 is the predominant chemokine secreted from IgE-activated human and mouse mast cells in vitro, colocalizes to mast cells in lung biopsies, and is elevated in asthmatic airways. CCR8, the receptor for CCL1, is expressed by approximately 70% of CD4(+) T lymphocytes recruited to the asthmatic airways, and the number of CCR8-expressing cells is increased 3-fold in the airways of asthmatic subjects compared with normal volunteers. In vivo, CCL1 expression in the lung is reduced in mast cell-deficient mice after aeroallergen provocation. Neutralization of CCL1 or CCR8 deficiency results in reduced mucosal lung inflammation, airway hyperresponsiveness, and mucus hypersecretion to a similar degree as detected in mast cell-deficient mice. Adenoviral delivery of CCL1 to the lungs of mast cell-deficient mice restores airway hyperresponsiveness, lung inflammation, and mucus hypersecretion to the degree observed in wild-type mice. The consequences of CCR8 deficiency, including a marked reduction in Th2 cytokine levels, are comparable with those observed by depletion of CD4(+) T lymphocytes. Thus, mast cell-derived CCL1- and CCR8-expressing CD4(+) effector T lymphocytes play an essential role in orchestrating lung mucosal inflammatory responses.


Real-time qualitative PCR for 57 human adenovirus types from multiple specimen sources.
Human adenoviruses (HAdVs) are ubiquitous double-stranded DNA viruses that cause a wide array of diseases in humans including pharyngitis, pneumonia, gastroenteritis, hemorrhagic cystitis, and keratoconjunctivitis. They also cause life-threatening opportunistic infections in immunocompromised individuals and are responsible for outbreaks in certain populations. Diagnosis is traditionally by cell culture or antigen detection methods. However, some HAdVs can take up to 4 weeks to isolate, and diarrheagenic types 40 and 41 will not grow in routine cell culture. Therefore, the goal of this study was to design a rapid, real-time PCR assay to detect all known 57 HAdV types from multiple different specimen sources. Primers and fluorescence resonance energy transfer hybridization probes were designed to target a 185-bp region of the penton base gene of HAdV. The analytical sensitivity was determined to be 10 copies/μl for HAdV types showing exact primer/probe homology in specimen matrix. Using whole-virus strains, the analytical sensitivity for representative HAdV types ranged from 10(-1) to 10(3) 50% tissue culture infective dose (TCID(50))/ml. The assay demonstrated 100% sensitivity and 99% specificity. This real-time PCR assay provides a rapid method for the detection of all 57 known HAdV types from respiratory specimens, blood, stool, urine, and ocular swabs.


Cyclooxygenase-2--specific inhibitors and cardiorenal function: a randomized, controlled trial of celecoxib and rofecoxib in older hypertensive osteoarthritis patients.
Arthritis and hypertension are common comorbid conditions affecting elderly adults. Use of nonsteroidal anti-inflammatory drugs in patients treated with antihypertensive medication can lead to destabilization of blood pressure control and other cardiorenal events. The potential for similar interactions with cyclooxygenase-2-specific inhibitors has not been fully explored. The authors evaluated the cardiorenal safety of two new cyclooxygenase-2-specific inhibitors, celecoxib and rofecoxib. This study was a 6-week, randomized, parallel-group, double-blind trial in patients with osteoarthritis who were > or =65 years of age and were taking antihypertensive agents. Patients received once-daily celecoxib 200 mg or rofecoxib 25 mg. The primary endpoints were the development of edema, changes in systolic blood pressure, and changes in diastolic blood pressure as measured at any time point in the study. Measurements occurred at baseline and after 1, 2, and 6 weeks of treatment. Eight hundred ten patients received study medication (celecoxib, n = 411; rofecoxib, n = 399). Nearly twice as many rofecoxib- compared with celecoxib-treated patients experienced edema (9.5% vs. 4.9%, P = 0.014). Systolic blood pressure increased significantly in 17% of rofecoxib- compared with 11% of celecoxib-treated patients (P = 0.032) at any study time point. Diastolic blood pressure increased in 2.3% of rofecoxib- compared with 1.5% of celecoxib-treated patients (P = 0.44). At week 6, the change from baseline in mean systolic blood pressure was +2.6 mmHg for rofecoxib compared with -0.5 mmHg for celecoxib (P = 0.007). Patients taking antihypertensive therapy and receiving cyclooxygenase-2-specific inhibitors should be monitored for the development of cardiorenal events. Patients receiving celecoxib experienced less edema and less destabilization of blood pressure control compared with those receiving rofecoxib.


Right heart overload contributes to cardiac natriuretic hormone elevation in patients with heart failure.
Atrial and brain natriuretic peptides (ANP and BNP) plasma concentration increases and holds a prognostic significance in patients with left ventricular dysfunction. We assessed the hypothesis that right ventricular (RV) overload might significantly contribute to plasma elevation of cardiac natriuretic hormones in patients with heart failure. Forty-one patients with cardiomyopathy and depressed left ventricular (LV) function (ejection fraction, EF, <40%), underwent cardiac magnetic resonance imaging (MRI) and resting plasma determination of ANP and BNP. Nineteen healthy subjects were also studied as control group. Ventricular volumes and function were assessed by MRI. In the group of patients, LVEF was 22.6+/-1.2% (controls: 61.2+/-1.3%, P<0.001, mean+/-S.E.M.), while RVEF was 48.2+/-2.5% (controls: 66.7+/-1.6%, P<0.001); LV and RV end diastolic/systolic volumes, corrected by body surface area, were 143+/-7/114+/-7 ml/m2 (controls 70+/-3/27+/-2 ml/m2, both P<0.001) and 66+/-3/37+/-4 ml/m2 (controls: 63+/-4/21+/-2 ml/m2, P<0.01 only for end-systolic volume). BNP plasma value was on average 324+/-39 pg/ml (range: 23-1280, controls 10+/-2 pg/ml), ANP value was 144+/-17 pg/ml (range: 26-534, controls 15+/-1 pg/ml). BNP positively correlated with either end-diastolic or end-systolic RV volume in patients, less with LV systolic, and not with LV diastolic volume. Moreover, a significant negative correlation was observed between BNP and either LVEF or RVEF. Conversely, ANP showed a significant correlation only with end-systolic RV volume and with both RVEF and LVEF. When multivariate stepwise linear regression analysis was applied LVEF resulted the only independent predictor for ANP plasma values (R=0.591, P<0.001), while LVEF and RV end-diastolic volume for BNP (R=0.881, P<0.001, and R=0.881, P=0.035, respectively). Right heart overload contributes independently to plasma elevation of natriuretic peptides. RV involvement, which is known to independently worsen prognosis in patients with cardiomyopathy, might contribute to their established prognostic power, inducing compensatory secretion of plasma cardiac natriuretic hormones.


Parsing genetically influenced risk pathways: genetic loci impact problematic alcohol use via externalizing and specific risk.
Genome-wide association studies (GWAS) identify genetic variants associated with a trait, regardless of how those variants are associated with the outcome. Characterizing whether variants for psychiatric outcomes operate via specific versus general pathways provides more informative measures of genetic risk. In the current analysis, we used multivariate GWAS to tease apart variants associated with problematic alcohol use (ALCP-total) through either a shared risk for externalizing (EXT) or a problematic alcohol use-specific risk (ALCP-specific). SNPs associated with ALCP-specific were primarily related to alcohol metabolism. Genetic correlations showed ALCP-specific was predominantly associated with alcohol use and other forms of psychopathology, but not other forms of substance use. Polygenic scores for ALCP-total were associated with multiple forms of substance use, but polygenic scores for ALCP-specific were only associated with alcohol phenotypes. Polygenic scores for both ALCP-specific and EXT show different patterns of associations with alcohol misuse across development. Our results demonstrate that focusing on both shared and specific risk can better characterize pathways of risk for substance use disorders. Parsing risk pathways will become increasingly relevant as genetic information is incorporated into clinical practice.


HLA-DR restriction in suppression of hematopoiesis by T cells from allogeneic bone marrow transplants.
Three allogeneic bone marrow transplantation patients who exhibited a suppressive subset of T cells for in vitro hematopoiesis have been investigated to determine whether this T cell suppressive effect was genetically restricted. In the three cases, T cells separated by sheep red cell rosetting inhibited blood colony-forming units granulocyte-monocyte (CFU-GM) and burst-forming unit erythroid (BFU-E) growth from the patients and from the bone marrow donors who were HLA identical, but not from randomly chosen unrelated subjects. In one case, cocultures were performed between the patient T cells and the T-depleted cells from eight siblings and from the mother. A marked inhibition (30 to 60%) of CFU-GM and BFU-E growth was found in the relatives who shared a haplo-identical HLA-DR 5. The same degree of suppression was found with respect to whether the siblings were homozygous or heterozygous for the HLA-DR 5 antigen, and whether or not they shared common class I antigens. This inhibition was totally abolished when a monoclonal antibody against HLA-DR was added, whereas a monoclonal antibody against class I histocompatibility antigen had no effect. To additionally demonstrate that this inhibition was mediated by a single HLA-DR haplotype, T cells from the patient were co-cultured with cells from three normal unrelated individuals, one with a phenotypically identical DR and two with only one haploidentical DR. Inhibition was similarly found in the subject exhibiting complete DR identity, and the subject with only the DR 5 haploidentical phenotype. These results demonstrate that a unique subset of T cells present in allogeneic bone marrow transplants specifically suppress differentiation of hemopoietic progenitors that bear one phenotypically haplo-identical HLA-DR antigen.


A prospective study of postoperative lymphedema after surgery for cervical cancer.
Lymphedema is a severe postoperative complication in oncological surgery. Multifrequency bioelectrical impedance analysis (MFBIA) is a new method for early lymphedema detection. The objective was to establish the methodology of MFBIA for lower-limb lymphedema and to detect a lymphedema in patients undergoing cervical cancer surgery. From a population of 60 patients undergoing cervical cancer surgery, 39 underwent radical hysterectomy Wertheim III (RAD group), and 21 underwent conservative surgery (laparoscopic lymphadenectomy plus simple trachelectomy/simple hysterectomy--CONS group). A control group of 29 patients (CONTR group) was used to determine the SD of impedance at zero frequency (R0). Patients were examined before surgery and at 3 and 6 months after surgery by MFBIA and by measuring the circumference of the lower limbs. No differences were found between the CONS and RAD groups on age, height, weight, and histopathologic type of tumor. However, the number of dissected lymph nodes differed significantly between the groups (17.3 in the CONS group vs 25.8 in the RAD group, P = 0.0012). The SD of R0 in the CONTR group was 36.0 and 39.0 for the right and the left leg, respectively. No difference in prevalence of lymphedema based on circumference method was found (35.9% in the RAD and 47.6% in the CONS groups, not statistically significant). No difference in the prevalence of lymphedema was found between the CONS and RAD groups. A methodology for MFBIA for the detection of lower-limb lymphedema was described.


Multiple interactions between maternally-activated signalling pathways control Xenopus nodal-related genes.
We have investigated the induction of the six Xenopus nodal-related genes, Xnr1-Xnr6, by maternal determinants. The beta-catenin pathway was modelled by stimulation using Xwnt8, activin-like signalling was modelled by activin, and VegT action was studied by overexpression in animal cap explants. Combinations of factors were examined, and previously unrecognised interactions were revealed in animal caps and whole embryos. For the induction of Xnr5 and Xnr6 in whole embryos, using a beta-catenin antisense morpholino oligonucleotide or a dominant negative XTcf3, we have demonstrated an absolute permissive requirement for the beta-catenin/Tcf pathway, in addition to the requirement for VegT action. In animal caps Xnr5 and Xnr6 are induced in response to VegT overexpression, and this induction is dependent upon the concomitant activation of the beta-catenin pathway that VegT initiates in animal caps. For the induction of Xnr3, VegT interacts negatively so as to inhibit the induction otherwise observed with wnt-signalling alone. The negative effect of VegT is not the result of a general inhibition of wnt-signalling, and does not result from an inhibition of wnt-induced siamois expression. A 294 bp proximal promoter fragment of the Xnr3 gene is sufficient to mediate the negative effect of VegT. Further experiments, employing cycloheximide to examine the dependence of Xnr gene expression upon proteins translated after the mid-blastula stage, demonstrated that Xnrs 4, 5 and 6 are 'primary' Xnr genes whose expression in the late blastula is solely dependent upon factors present before the mid-blastula stage.


Treatment of Femur Fractures in Preschool Children with Double Pin Technique: Immediate Incorporated Hip Spica Casting by Two K-Wires.
The immediate hip spica casting is a popular treatment method for femoral fractures in the pediatric age group. Femoral shortening is the unacceptable result for the treatment. In this technique, we tried to describe the immediate spica casting technique pinning with double K-wire in preschool children and evaluate the results with this method. Sixteen patients (ten boys and six girls) are treated with this technique. Patients were treated under general anesthesia in the operating room. K-wire was introduced in the supracondylar area of the femur passing from the lateral to the medial side through both cortexes, and removed from the skin in the medial section. A second K-wire was introduced through the lateral side of the proximal fragment. While an assistant was applying traction to the pin to maintain the reduction, the lower extremity was put in a spica cast on the fractured side, while the knee joint on the other side was excluded from the spica casting process. The angulation after hip spica cast applied was 9.1° (range 7°-12°) on the frontal plane and 8.9° (range 5°-17°) on the sagittal plane. After spica cast removal, it was 8.5° (range 5°-13°) on the frontal plane and 9° (range 5°-17°) on the sagittal plane. No significant difference was found. The final mean leg length discrepancy was 4.8 mm (range 0-10 mm) at the time of cast removal. This technique may be effective for femoral diaphyseal fractures in preschool children to combine the spirits of external fixation with hip spica casting in one construct in order to correct angular deformities and length discrepancies.


Flow cytometry in impression cytology specimens. A new method for evaluation of conjunctival inflammation.
To investigate feasibility and potential uses of flow cytometry in impression cytology as a new procedure to assess and quantify conjunctival inflammation. Specimens for cytology were collected by impression from 30 patients with various chronic ocular surface disorders and from 10 normal subjects. Two specimens were obtained in each eye: One was transferred onto a glass slide and processed by immunofluorescence with antibodies to human leukocyte antigen (HLA)-DR antigens; cells from the other were suspended in phosphate-buffered saline for flow cytometry. Monoclonal antibodies to HLA-DR antigens and CD23, the low affinity receptor to immunoglobulin E, were used. Abnormal expression of HLA-DR and CD23 by conjunctival cells was found in 13 of 18 dry eyes and in 20 of 22 eyes with chronic conjunctivitis, whereas specimens remained almost negative (less than 10% of cells were positive) in normal eyes. Percentages of positive cells ranged between 20% and 98% of all conjunctival cells. Correlation between the two methods, immunocytology and flow cytometry, was highly significant (coefficient of correlation 0.77, P = 0.0001). Moreover, HLA-DR positivity, at its strongest intensity, was observed in a minority of cells (1% to 12%), most of which were resident class II-expressing dendritic cells. Percentages of those cells expressing high levels of HLA-DR were 3 +/- 1.2% in normal eyes, 5.8 +/- 4% in dry eyes (P = 0.05), and 5.9 +/- 3.5% in eyes with chronic conjunctivitis (P = 0.02). Results of this preliminary study confirm that conjunctival epithelial cells may abnormally express inflammatory markers in chronic ocular surface disorders. Development of flow cytometry in analysis of cytologic specimens provides a new, sensitive, and objective tool for exploring conjunctival pathology.


Autoantibodies in scleroderma and polymyositis: an update.
Cutaneous manifestations of systemic connective tissue diseases, such as scleroderma and its variants, polymyositis, and dermatomyositis, often prompt early dermatologic consultation. Indirect immunofluorescent autoantibody determinations using tissue culture substrates are initial screening tests that are highly positive in the majority of patients with scleroderma and its variants, but are less frequently positive in patients with polymyositis and dermatomyositis. When combined with second-level analyses for the multiplicity of precipitin autoantibodies that have been defined in both these major classes of rheumatic diseases, most autoantibodies of both diagnostic and prognostic significance can be defined efficiently and cost-effectively. The major autoantibody specificities characteristic of these connective tissue diseases are summarized in this article, with emphasis on current concepts of their clinical molecular, and possible pathogenetic significance.


Mesolimbic dopamine neurons in the brain reward circuit mediate susceptibility to social defeat and antidepressant action.
We previously reported that the activity of mesolimbic dopamine neurons of the ventral tegmental area (VTA) is a key determinant of behavioral susceptibility vs resilience to chronic social defeat stress. However, this was based solely on ex vivo measurements, and the in vivo firing properties of VTA dopamine neurons in susceptible and resilient mice, as well as the effects of antidepressant treatments, remain completely unknown. Here, we show that chronic (10 d) social defeat stress significantly increased the in vivo spontaneous firing rates and bursting events in susceptible mice but not in the resilient subgroup. Both the firing rates and bursting events were significantly negatively correlated with social avoidance behavior, a key behavioral abnormality induced by chronic social defeat stress. Moreover, the increased firing rates, bursting events, and avoidance behavior in susceptible mice were completely reversed by chronic (2 week), but not acute (single dose), treatments with the antidepressant medication fluoxetine (20 mg/kg). Chronic social defeat stress increased hyperpolarization-activated cation current (I(h)) in VTA dopamine neurons, an effect that was also normalized by chronic treatment with fluoxetine. As well, local infusion of I(h) inhibitors ZD7288 (0.1 μg) or DK-AH 269 (0.6 μg) into the VTA exerted antidepressant-like behavioral effects. Together, these data suggest that the firing patterns of mesolimbic dopamine neurons in vivo mediate an individual's responses to chronic stress and antidepressant action.


Determining corneal power using Pentacam after myopic photorefractive keratectomy.
To assess the accuracy of Pentacam Scheimpflug camera for corneal power measurement in eyes with previous photorefractive keratectomy for myopia. In this comparative interventional case series, 35 eyes of 35 patients who had myopic photorefractive keratectomy were studied. Corneal power was measured by conventional topography and Pentacam Scheimpflug camera, and equivalent keratometry readings (EKR) in different central corneal rings (0.5 to 4.5 mm), true net power and simulated keratometry (K) measurements as well as those obtained using Shammas no-history, Koch-Maloney and Haigis methods were compared with clinical history method. All corneal power measurements except for the topography simulated K and true net power values were statistically similar to the clinical history values. Simulated keratometry and 4.5-mm EKR values were more closely correlated with clinical history method. Shammas formula, Pentacam simulated K and 3-, 4- and 4.5-mm EKR provided a 95% confidence interval within +/-0.50 D of the mean clinical history method value, among these, the width of the 95% limits of agreement (LoA) was narrower for Shammas and Pentacam simulated K and 4.5-mm EKR values; however, considerably large 95% LoA were found between each of these values and those obtained with the clinical history method. Estimated preoperative keratometry was statistically similar to the preoperative measurement; however, estimated refractive change was different from actual value. The Pentacam 4.5-mm EKR and simulated keratometry may be used as an alternative to clinical history method to predict corneal power when pre-keratorefractive surgery data are unavailable; however, wide LoA should be considered in the calculations.


The role of school physical education on adolescents' fitness levels during the pandemic period from COVID-19: An observational study of the Italian scientific high school-section sport and physical activity.
In Italy, in 2013, a new school curriculum with a sport character was established in high schools, called Sports High School (SHS). The aims of this study were: (1) to assess the fitness levels of SHS students who, respecting all the safety rules to limit the spread of COVID-19, practiced physical education (PE) at school with continuity for almost all of 2021, and to compare them with Traditional High School (THS) students; (2) to evaluate as the SHS may have influenced the fitness levels in adolescents. This is a case-control study in which thirty participants were enrolled (SHS: n = 15; THS: n = 15). To assess the fitness levels, the following tests were administered: the Static Baropodometric and Stabilometric Analyses, the Counter Movement Jump (CMJ), and the Handgrip test. All these tests were administered when the non-pharmaceutical interventions (NPIs) for COVID-19 allowed the resumption of PE lessons (T0) and 2 months after their resumption (T1). Unpaired t-test between SHS (T0) vs. THS (T0) showed significant differences between: Handgrip test Dx and Handgrip test Sx (both p < 0.001), Surface Sx foot and Surface Dx foot (both p < 0.05), and CMJ (p < 0.001). These results were also confirmed in T1. The performance analysis between T0 and T1 of both SHS and THS showed improvements in SHS: Handgrip test Dx (p < 0.05; d = 0.57), Handgrip test Sx (p < 0.01; d = 0.87), and CMJ (p < 0.05; d = 0.59). Pearson's analysis of the results of the tests showed significant strong correlations between: Handgrip test Dx and Handgrip test Sx (R = 0.959; p < 0.001), Handgrip test Dx and CMJ (R = 0.881; p < 0.001), Handgrip test Sx and CMJ (R = 0.893; p < 0.001). The same analysis showed significant but moderate correlations between: Surface foot Sx and CMJ (R = 0.489; p < 0.01), Surface foot Sx and y-mean (R = 0.485; p < 0.01), Surface foot Dx and CMJ (R = 0.444; p < 0.05). This study is in agreement with the literature showing that the quarantine period and the NPIs for COVID-19 caused a decrease in fitness levels in adolescents. Our results showed that students of SHS recorded higher strength performance both in the Handgrip test and in the CMJ.


Physical activity and autoimmune diseases: Get moving and manage the disease.
Physical activity, by definition, is any skeletal muscle body movement that results in energy expenditure. In the last few decades, a plethora of scientific evidences have accumulated and confirmed the beneficial role of physical activity as a modifiable risk factor for a wide variety of chronic diseases including cardiovascular diseases (CVDs), diabetes mellitus and cancer, among others. Autoimmune diseases are a heterogeneous group of chronic diseases, which occur secondary to loss of self-antigen tolerance. With the advent of biological therapies, better outcomes have recently been noted in the management of autoimmune diseases. Nonetheless, recent research highlights the salient role of modifiable behaviors such as physical inactivity on various aspects of the immune system and autoimmune diseases. Physical activity leads to a significant elevation in T-regulatory cells, decreased immunoglobulin secretion and produces a shift in the Th1/Th2 balance to a decreased Th1 cell production. Moreover, physical activity has been proven to promote the release of IL-6 from muscles. IL-6 released from muscles functions as a myokine and has been shown to induce an anti-inflammatory response through IL-10 secretion and IL-1β inhibition. Physical activity has been shown to be safe in most of autoimmune diseases including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), inflammatory bowel diseases (IBD), as well as others. Additionally, the incidence of RA, MS, IBD and psoriasis has been found to be higher in patients less engaged in physical activity. As a general trend, patients with autoimmune diseases tend to be less physically active as compared to the general population. Physically active RA patients were found to have a milder disease course, better cardiovascular disease (CVD) profile, and improved joint mobility. Physical activity decreases fatigue, enhances mood, cognitive abilities and mobility in patients with MS. In SLE patients, enhanced quality of life and better CVD profile were documented in more physically active patients. Physically active patients with type 1 diabetes mellitus have a decreased risk of autonomic neuropathy and CVD. Both fibromyalgia and systemic sclerosis patients report decreased disease severity, pain, as well as better quality of life with more physical activity. Further, SSc patients improve their grip strength, finger stretching and mouth opening with increased level of exercise. The purpose of this paper is to review the clinical evidence regarding the safety, barriers to engagement, and impact of physical activity on autoimmune diseases.


Clinical management of imported malaria in Italy: results from a national cross-sectional survey in 2015.
In Italy, malaria continues to be one of the most common imported parasitoses; therefore, continuous surveillance of epidemiological data and clinical management is needed. In 2016, the National Institute for Infectious Diseases 'Lazzaro Spallanzani' in Rome promoted a retrospective questionnaire-based survey to assess the clinical management of imported malaria cases in Italy in 2015. The questionnaire was sent to 104 Tropical and/or Infectious Diseases Units in the country, and 37 of them filled out and returned the questionnaires. A total of 399 malaria cases were reported in 2015, mostly caused by Plasmodium falciparum and imported from Africa. Malaria chemoprophylaxis was correctly used by a minority of patients. Most patients presented with uncomplicated malaria and were treated orally. In severe cases, intravenous artesunate or quinine alone or in combination were administered, although one third of these severe cases received oral treatment. This retrospective survey reveals a lack of homogeneity in management of malaria-imported cases in Italy. Improvement of malaria chemoprophylaxis, standardization of clinical management of malaria cases and harmonization of oral and intravenous drug availability are needed throughout the country.


[Arachnoiditis ossificans].
Arachnoiditis ossificans is a uncommon disorder, is generally associated with neurologic deficits, but patients could be only mild sintomatic, regardless of the degree of ossification. Unenhanced CT has been well show to be sensitive for the disorder, useful to evaluate the full extenstion of the ossified mass and their remotion if surgical intervention is needed. On MRI imaging the manifestations could be minimal and variable, being important to access another pathologic conditions (arachnoid cysts, intramedullary cysts and syringomyelia).


Endoscopy management algorithms: role of cyanoacrylate glue injection and self-expanding metal stents in acute variceal haemorrhage.
Mortality from acute variceal bleeding (AVB) has improved markedly over the last 2-3 decades due to increased specialisation and standardisation of medical and endoscopic practice culminating in the production of consensus guidance based on expert opinion. Nonetheless, despite greater exposure, training and endoscopic practices, 30-day mortality still remains high at around 30%. This is a reflection of the high morbidity with liver disease, and limited endoscopic experience and/or endoscopic techniques used by the majority of general endoscopists. Clinical necessity defines our drive for further endoscopic innovation to improve 'best practice' and, therefore, clinical outcomes accordingly. Sclerotherpy, variceal band ligation and/or rescue balloon tamponade have been entrenched in most treatment algorithms over the decades. However, in recent years and albeit limited to specialised liver centres, cyanoacrylate glue injection therapy (for oesophageal and gastric varices), and the placement of a self-expanding metallic stent for oesophageal varices have begun to offer improved endoscopic care in experienced hands. Yet even in specialised centres, their application is sporadic and operator dependent. Here, we discuss the evidence of these newer endoscopic approaches, and hope to propose their inclusion in endoscopic therapy algorithms for 'best practice' management of AVB in all appropriately supported endoscopy units.


The thigh may not be suitable as an injection site for patients self-injecting sumatriptan.
The objective of our study was to evaluate the differences between injection sites in the midthigh and the upper lateral quadrant of the gluteal area regarding the effect, depth of subcutaneous tissue, side effects, and patient preference in patients with cluster headache who self-inject 6 mg (0.5 ml) of sumatriptan. Our open, prospective clinical study was performed at the outpatient department of a Swedish university clinic. There were 19 male and one female subjects, 34 to 68 years old, suffering from cluster headache. We measured the subcutaneous tissue depth by ultrasound. Subjects performed four self-injections of sumatriptan, two in the thigh and two in the gluteal area. We evaluated the subcutaneous tissue depth from the skin surface to the muscle fascia, the effect on headache, side effects, and patient preference regarding the injection site. Subcutaneous tissue depth laterally in the thigh was 2 to 12 mm (median, 4 mm) and in the gluteal area was 34 to 68 mm (median, 45 mm). The needle of the self-injector protrudes 5 to 6 mm. Forty thigh and 39 gluteal injections were recorded. The effect on headache was equal. Following injection in thigh the patients experienced more bleeding (p < 0.001, chi 2); local pain (p < 0.05, chi 2); and a feeling of oppression in the head, neck, and chest area (p < 0.05, chi 2); compared with injections in the gluteal area. Fifteen patients preferred the gluteal area as the injection site after the study, two patients had no preference, and three preferred the thigh. When using the self-injector in the lateral aspect of the thigh, intramuscular injection is liable to occur frequently in male patients. This may explain the differences in local and general side effects observed in this study. The upper lateral quadrant of the gluteal area is a more suitable injection site for male, and some female, patients when using the sumatriptan self-injector.


Low incidence of HIV infection and decreasing incidence of sexually transmitted infections among PrEP users in 2020 in Germany.
Objectives of this study, as part of a nation-wide HIV pre-exposure prophylaxis (PrEP) evaluation project, were to determine the incidence of infections with HIV, chlamydia, gonorrhea, syphilis, hepatitis A/B/C in persons using PrEP, and to describe the health care funded PrEP use in Germany. Additionally, factors associated with chlamydia/gonorrhea and syphilis infections were assessed. Anonymous data of PrEP users were collected at 47 HIV-specialty centers from 09/2019-12/2020. Incidence rates were calculated per 100 person years (py). Using longitudinal mixed models, we analyzed risk factors associated with sexually transmitted infections (STIs). 4620 PrEP users were included: 99.2% male, median age 38 years (IQR 32-45), 98.6% men who have sex with men (MSM). The median duration of PrEP exposure was 451 days (IQR 357-488), totaling 5132 py. Four HIV infections were diagnosed, incidence rate 0,078/100py (95% CI 0.029-0.208). For two, suboptimal adherence was reported and in the third case, suboptimal adherence and resistance to emtricitabine were observed. One infection was likely acquired before PrEP start. Incidence rates were 21.6/100py for chlamydia, 23.7/100py for gonorrhea, 10.1/100py for syphilis and 55.4/100py for any STI and decreased significantly during the observation period. 65.5% of syphilis, 55.6% of chlamydia and 50.1% of gonorrhea cases were detected by screening of asymptomatic individuals. In a multivariable analysis among MSM younger age, PrEP start before health insurance coverage and daily PrEP were associated with greater risk for chlamydia/gonorrhea. Symptom triggered testing and a history of STI were associated with a higher risk for chlamydia/gonorrhea and syphilis. A significantly lower risk for chlamydia/gonorrhea and syphilis was found for observations during the COVID-19 pandemic period. We found that HIV-PrEP is almost exclusively used by MSM in Germany. A very low incidence of HIV infection and decreasing incidence rates of STIs were found in this cohort of PrEP users. The results were influenced by the SARS-CoV-2 pandemic. Rollout of PrEP covered by health insurance should be continued to prevent HIV infections. Increased PrEP availability to people at risk of HIV infection through the elimination of barriers requires further attention. Investigation and monitoring with a longer follow-up would be of value.


Modelling Behaviour of a Carbon Epoxy Composite Exposed to Fire: Part I-Characterisation of Thermophysical Properties.
Thermophysical properties of a carbon-reinforced epoxy composite laminate (T700/M21 composite for aircraft structures) were evaluated using different innovative characterisation methods. Thermogravimetric Analysis (TGA), Simultaneous Thermal analysis (STA), Laser Flash analysis (LFA), and Fourier Transform Infrared (FTIR) analysis were used for measuring the thermal decomposition, the specific heat capacity, the anisotropic thermal conductivity of the composite, the heats of decomposition and the specific heat capacity of released gases. It permits to get input data to feed a three-dimensional (3D) model given the temperature profile and the mass loss obtained during well-defined fire scenarios (model presented in Part II of this paper). The measurements were optimised to get accurate data. The data also permit to create a public database on an aeronautical carbon fibre/epoxy composite for fire safety engineering.


The epidemiology of acute myocardial infarction and ischemic heart disease in Canada: data from 1976 to 1991.
To present national trends in mortality rates for myocardial infarction and cardiovascular disease. Observational study using mortality statistics and hospital separation data from Statistics Canada for the period 1976 to 1991. Despite ageing of the population, there has been a substantial decrease in the number of deaths attributed to ischemic heart disease, from 51,000 in 1976 to 44,000 in 1991, with most of the decrease due to fewer deaths from myocardial infarction. Although age-adjusted death rates remain higher for men, the observed mortality decline has been more pronounced in men than in women. Age-adjusted separation rates have also decreased, suggesting a decrease in the incidence of myocardial infarction, particularly in the 45 to 64 year age group. The duration of hospital stay has shortened dramatically. From 1976 to 1991, mortality rates for ischemic heart disease in Canada decreased sharply, suggesting that advancements observed in clinical trials are being translated to the population level. The decrease appears to be due to both preventive measures and improved hospital care, but further studies are necessary to define better the relative contribution of each factor. The extent of this progress over the past 15 years is similar to the American experience.


In Vitro and In Silico Studies for the Identification of Potent Metabolites of Some High-Altitude Medicinal Plants from Nepal Inhibiting SARS-CoV-2 Spike Protein.
Despite ongoing vaccination programs against COVID-19 around the world, cases of infection are still rising with new variants. This infers that an effective antiviral drug against COVID-19 is crucial along with vaccinations to decrease cases. A potential target of such antivirals could be the membrane components of the causative pathogen, SARS-CoV-2, for instance spike (S) protein. In our research, we have deployed in vitro screening of crude extracts of seven ethnomedicinal plants against the spike receptor-binding domain (S1-RBD) of SARS-CoV-2 using an enzyme-linked immunosorbent assay (ELISA). Following encouraging in vitro results for Tinospora cordifolia, in silico studies were conducted for the 14 reported antiviral secondary metabolites isolated from T. cordifolia-a species widely cultivated and used as an antiviral drug in the Himalayan country of Nepal-using Genetic Optimization for Ligand Docking (GOLD), Molecular Operating Environment (MOE), and BIOVIA Discovery Studio. The molecular docking and binding energy study revealed that cordifolioside-A had a higher binding affinity and was the most effective in binding to the competitive site of the spike protein. Molecular dynamics (MD) simulation studies using GROMACS 5.4.1 further assayed the interaction between the potent compound and binding sites of the spike protein. It revealed that cordifolioside-A demonstrated better binding affinity and stability, and resulted in a conformational change in S1-RBD, hence hindering the activities of the protein. In addition, ADMET analysis of the secondary metabolites from T. cordifolia revealed promising pharmacokinetic properties. Our study thus recommends that certain secondary metabolites of T. cordifolia are possible medicinal candidates against SARS-CoV-2.


Implementing Surviving Sepsis guidelines in a district general hospital.
We describe efforts made at Blackpool Victoria Hospital to implement the well-established international Surviving Sepsis guidelines. These included posters, pocket guides and stickers inserted in patient notes. All doctors and nurses in acute areas received specific information and education. Sepsis teams comprising both doctors and nurses were formed to encourage implementation and to audit and disseminate data. Data collection occurred from February to November 2009. Cases were considered prospectively at the time of initial assessment and 198 patients were identified; 169 (85%) had blood cultures taken; 146 (74%) had lactate levels measured; and 145 (74%) received antibiotics within the target time. We believe these results demonstrate relatively effective implementation of guidelines in the challenging environment of a district general hospital. Our results could be replicated easily and provide a good way of reducing patient mortality at minimal financial cost.


Bladder Cancer Metastasis to the Breast in a Male Patient: Imaging Findings on Mammography and Ultrasonography.
Male breast cancer is rare, accounting for approximately 1% of breast cancers. Metastasis from extra-mammary malignancy to the breast in men is extremely rare. The most common primary tumors that metastasize to the breast in male are prostate, lung, stomach, colorectal cancer, melanoma, and sarcoma. To our knowledge, only a few cases of bladder cancer presenting with metastasis to the male breast have been reported, and metastasis with infiltration rather than mass is extremely rare. We report imaging findings on mammography and ultrasonography in a 59-year-old male with bladder cancer metastatic to the breast.


Solution properties of perfluorinated anionic surfactants with divalent counterion of separate electric charges.
Novel surfactants of perfluorinated double long-chain salts with divalent counterion of separate electric charge, 1,1-(1,omega-alkanediyl)bispyridinium diperfluorononanoate (CnBP(FC9)2, n = 2, 4, 6, 8) were newly synthesized. Their solution properties were investigated by surface tension measurement over the temperature range from 298.2 to 313.2 K, where magnesium diperfluorononanoate (Mg(FC9)2) was employed as a reference surfactant with divalent counterion of concentrated electric charge. From change of surface tension with concentration, the critical micelle concentration (CMC), surface excess (Gamma), apparent molecular surface area (A), and -log(concentration to reduce surface tension of water by 20 mN m-1) (pC20) were determined. The CMC values of CnBP(FC9)2 decreased with increasing charge separation and with increasing temperature, where the values of CnBP(FC9)2 were much smaller than those of Mg(FC9)2. In addition, the pC20 values of the former were also much larger than those of the latter. These results indicate a strong influence of the extent of charge separation or the spacer length of the counterions upon surface activity of the fluorinated surfactants. The surface excess or the corresponding apparent molecular surface area monotonously changed with the spacer length (n < or = 6), whereas the behavior for n = 8 was much different from the other CnBP(FC9)2 due to conformational change in the in-between alkanediyl chain. The entropy changes (Deltas) for the surface adsorption or condensation were found to be mostly negative for CnBP(FC9)2, where the changes approached zero with an increase in the charge separation. On the other hand, the changes for Mg(FC9)2 were positive over the whole concentration below the CMC. In addition, Brewster angle microscopy indicated no condensation of the present surfactants just at the air/solution interface.


Biological activation of zirconia surfaces by chemical modification with IGF-1.
The purpose of this study was to improve the adhesion and extension of human gingival epithelial cells (HGECs) to the yttria-stabilized zirconia polycrystal (Y-TZP) surfaces by immobilization of insulin-like growth factor 1 (IGF-1). Surface analyses by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) showed that IGF-1 was successfully immobilized on the Y-TZP surfaces. There was no significant difference between the number of cells attached to the IGF-1-immobilized Y-TZP surfaces and on the as-polished Y-TZP surfaces either at 3 or 72 h. However, IGF-1-immobilized Y-TZP surfaces yielded a significantly higher expression of integrin β4 mRNA and laminin-5 mRNA, and enhanced adhesion strength of HGECs after 72 h of incubation. There was no difference between the amount of adhered Streptococcus gordonii (S. gordonii) found on the IGF-1-immobilized Y-TZP surfaces and on the as-polished Y-TZP surfaces. These results suggested that the IGF-1-immobilized Y-TZP surfaces developed using the method reported herein enhanced the adhesion and extension of HGECs to the Y-TZP surfaces without enhancing S. gordonii adhesion.


Quality Measurement Gaps and Future Directions in the Assessment of Obesity.
The disease of obesity continues to increase in prevalence and severity yet obesity care, treatment, and coverage are scarce. Progress has been made in the development and implementation of quality measures in clinical practice and organizational performance. However, major gaps and limitations exist in the context of measuring guideline-based clinical care for obesity. Obesity quality measures have entered various stages of testing and development, but only a select few are included in reporting and payment programs. One process measure for adults, "Preventive Care and Screening: Body Mass Index (BMI) Screening and Follow-Up Plan" is used in Medicare. One process measure for pediatrics, "Weight Assessment and Counseling for Nutrition and Physical Activity in Children and Adolescence" is used in Medicare, health insurance plans, and Medicaid. No outcome or digital quality measure exists for the disease of obesity. One quality measure development project is underway that is testing the performance of four measures, including outcome measures for obesity. The general absence of quality measures for obesity means that there are no objective measures to address the quality of obesity care or its outcome. More meaningful efforts are needed to seriously integrate obesity with quality performance measurement and value in healthcare payment programs.


Changes in Anticoagulant Utilization Among United States Nursing Home Residents With Atrial Fibrillation From 2011 to 2016.
Background Nursing home residents with atrial fibrillation are at high risk for ischemic stroke and bleeding events. The most recent national estimate (2004) indicated less than one third of this high-risk population was anticoagulated. Whether direct-acting oral anticoagulant ( DOAC ) use has disseminated into nursing homes and increased anticoagulant use is unknown. Methods and Results A repeated cross-sectional design was used to estimate the point prevalence of oral anticoagulant use on July 1 and December 31 of calendar years 2011 to 2016 among Medicare fee-for-service beneficiaries with atrial fibrillation residing in long-stay nursing homes. Nursing home residence was determined using Minimum Data Set 3.0 records. Medicare Part D claims for apixaban, dabigatran, edoxaban, rivaroxaban, and warfarin were identified and point prevalence was estimated by determining if the supply from the most recent dispensing covered each point prevalence date. A Cochran-Armitage test was performed for linear trend in prevalence. On December 31, 2011, 42.3% of 33 959 residents (median age: 85; Q1 79, Q3 90) were treated with an oral anticoagulant, of whom 8.6% used DOAC s. The proportion receiving treatment increased to 47.8% of 37 787 residents as of December 31, 2016 ( P<0.01); 48.2% of 18 054 treated residents received DOAC s. Demographic and clinical characteristics of residents using DOAC s and warfarin were similar in 2016. Half of the 8734 DOAC users received standard dosages and most were treated with apixaban (54.4%) or rivaroxaban (35.8%) in 2016. Conclusions Increases in anticoagulant use among US nursing home residents with atrial fibrillation coincided with declining warfarin use and increasing DOAC use.


Analysis of cytokine gene expression during fetal thymic ontogeny using the polymerase chain reaction.
The expression of 12 cytokine and cytokine receptor genes during mouse thymic ontogeny has been studied using the polymerase chain reaction. The data reveal that IL-2 gene transcription does not begin in the thymus until fetal day (Fd) 16. However, IL-2R beta-chain mRNA was detectable throughout the period of investigation, from Fd 14 to 20. This suggests that the potential for high or intermediate affinity IL-2 binding is present in the thymus before the IL-2 gene is actually expressed. Overall, cytokine gene induction occurred in two distinct patterns. In the first group, represented by IL-1 beta, IL-4, IL-5, IL-6, IL-7, IFN-gamma, and lymphotoxin (lymphotoxin or TNF-beta) there was detectable mRNA present from Fd 14 to 20. IL-1 alpha, IL-2, IL-3, and GM-CSF followed a second pattern in which gene induction was delayed until Fd 16. The expression of IL-7 and IL-2 genes showed an interesting temporal relationship. IL-7 mRNA transcription occurred before IL-2 but diminished from Fd 16 after the onset of IL-2 gene induction. Because these data reflect the in vivo progression of gene expression during T cell ontogeny, they provide a guideline for assessing the physiologic significance of thymocyte responses to cytokines at different stages of fetal development.


Analysis of bile acids and their conjugates by capillary electrochromatography/electrospray ion trap mass spectrometry.
Different macroporous, monolithic capillary columns were prepared to separate various bile acid mixtures through capillary electrochromatography (CEC) at high efficiency. These columns are shown to be ideally suitable for coupling to an electrospray ionization/ion trap mass spectrometer. Detection and structural identification of different bile acid derivatives in either the positive- or negative-ion mode necessitated column technologies with different polarities and the capabilities of a reversed electroosmotic flow. High column efficiencies (610,000 theoretical plates/meter for glycocholic acid in normal-phase separation) were preserved in the coupling to mass spectrometry (MS), with the detection limits of approximately 40 femtomole (for cholic acid) and identification through CEC/MS/MS.


Early diagnosis of acute kidney injury in critically ill patients.
Acute kidney injury (AKI) is a common and serious problem in critically ill patients. Tests currently used to detect AKI (i.e., serum creatinine, serum urea and various urinary indices) often result in delayed detection of injury--becoming abnormal at 48-72 h after the initial insult. This delayed detection translates into a potential missed opportunity for therapeutic interventions at a time when kidney damage may be limitable or reversible. This may also, in particular, account for the poor clinical outcomes commonly associated with AKI. The development of novel serum and urinary biomarkers capable of detecting AKI at an earlier phase of illness is therefore vital. This article will review the pitfalls of current conventional testing in kidney injury and discuss the emergence of novel biomarkers with the potential to revolutionize the field of critical care nephrology.


Topical Application of TGF-β-Activating Peptide, KRFK, Prevents Inflammatory Manifestations in the TSP-1-Deficient Mouse Model of Chronic Ocular Inflammation.
Chronic inflammation of the ocular surface poses a risk of vision impairment. The understanding of the molecular mechanisms that are involved in the inflammatory response is critical to identify novel molecular targets. Recently, thrombospondin-1 (TSP-1) has emerged as a key player in ocular surface homeostasis that efficiently activates the TGF-β2 isoform that is predominantly expressed in the ocular mucosa. Here, the potential of the peptide derived from TSP-1 (KRFK), that can activate TGF-β, is proposed as a potentially applicable therapeutic for chronic ocular surface inflammatory disorders. Our in vitro results confirm that the chosen peptide activates TGF-β, reducing the expression of co-stimulatory molecules on dendritic cells, driving them towards a tolerogenic phenotype. For the in vivo studies, the TSP-1-/- mouse is used as a pre-clinical model of chronic ocular inflammation. We observe that the topical application of KRFK alters the peripheral balance of effectors by reducing the proportion of pathogenic Th1 and Th17 cells while increasing Treg cell proportion in cervical lymph nodes. In line with these findings, the development of chronic ocular surface inflammation is significantly prevented in KRFK-treated TSP-1-/- mice, as assessed by clinical parameters and inflammatory cytokine expression in conjunctival and lacrimal gland tissues. Together, our results identify the KRFK peptide as a novel therapeutic option to prevent the development of chronic inflammatory manifestations of the ocular surface.


[ApoE genotype influences on efficacy and safety of thrombolytic treatment for ischemic stroke].
Apolipoprotein E has been associated with intracerebral hemorrhages and with neurological outcome of ischemic stroke patients treated with rt-PA. Therefore, we hypothesized that ApoE genotype might influence the appearance of post-tPA hemorrhagic transformation and that the favorable outcome of E2 patients might be due to better rates of recanalization. We analyzed the ApoE genotype of 77 patients with ischemic stroke involving the territory of the middle cerebral artery who received rt- PA within 3 h of symptoms onset. The hemorrhagic events were evaluated by computed tomography and the arterial recanalization by transcraneal doppler. We did not observe any association between ApoE genotypes and the rates of hemorrhagic transformation following rt-PA treatment (E2 = 33.3 %, E3 = 32.7%, E4=11.8%; p=0.241). Rates of artery recanalization following thrombolysis were similar regarding ApoE genotypes at 1 hour post-tPA administration (E2=40%, E3+E4=45.9%; p=0.799), at 6 hours post-tPA (E2=80%, E3+E4=62.3 %; p=0.43) or at 24 hours post-tPA (E2= 100%, E3+E4=75%, p=0.203). No association was observed between ApoE genotypes and NIHSS scores at 48 hours (E2=8, E3+E4=12; p=0.811) nor with the modified ranking scale at 3 months (E2=1.5, E3+E4=4; p=0.350). In our group of stroke patients, ApoE genotypes are not related with the presence of hemorrhagic transformations neither with the rates of recanalization following thrombolytic treatment.


Comparison of tracheal tube cuff pressure with two technique: fixed volume and minimal leak test techniques.
Introduction: There is a correlation between endotracheal cuff pressure and airway complication; therefore, cuff pressure measurement is of an essential importance. The gold standard technique is measuring the cuff pressure by a calibrated manometer. However, there are several methods that injects air into balloon pilot and measures the cuff pressure. The aim of this study is to compare the tracheal cuff pressure measurement by two methods: fixed volume and minimal leak test (MLT). Methods: This descriptive study was performed at the emergency department on 110 patients. Patients were randomized into two groups. For one group, fixed volume technique and for the other group MLT was used. Results: Mean cuff pressure was 46.07±23.54 cmH2O in the fixed volume group and 33.72±9.14 cmH2O in the MLT group (P=0.05) which is significantly higher in the fixed volume group (P=0.028). In addition, 56.4% and 78.2% of the subjects had normal cuff pressure in the fixed volume group and MLT group, respectively; indicating a significantly higher rate in MLT group (P=0.025). Conclusion: Both techniques cause above normal intracuff pressure; however, MLT produces more acceptable pressure than fixed volume. It seems that the volume of 10 cc produces high pressures; therefore, fixed values may yield more appropriate results in lower volumes.


[The place of ultrasound diagnosis in obstetrics].
A review is presented of the diagnostic applications of the ultrasonic scan technique in obsterics. In the 1st trimester of pregnancy the demonstration of the gestational sac and of an embryo is most important. During the further course of pregnancy the growth of the gestational sac and motions of the fetus can be observed. A disturbance in the development of the pregnancy can be deduced from these parameters. The various types of abortion can be differentiated by means of the scan. In the 2nd trimester the biometry of the fetus is most important. The motion behaviour of the fetus seems to be a good indicator of its well-being. The location of the placenta and the evidence of twins are easily demonstrable. In the 3rd trimester the biometry of the fetus is a good parameter of its maturity. The diagnosis of the fetal position is important for the management of delivery. Apart from these applications of ultrasound in obstetrics, the procedure is of great value in the diagnosis of tumours and the measurement of the true pelvis in pregnancy.


Anisotropic Interfacial Force Field for Interfaces of Water with Hexagonal Boron Nitride.
This study introduces an anisotropic interfacial potential that provides an accurate description of the van der Waals (vdW) interactions between water and hexagonal boron nitride (h-BN) at their interface. Benchmarked against the strongly constrained and appropriately normed functional, the developed force field demonstrates remarkable consistency with reference data sets, including binding energy curves and sliding potential energy surfaces for various configurations involving a water molecule adsorbed atop the h-BN surface. These findings highlight the significant improvement achieved by the developed force field in empirically describing the anisotropic vdW interactions of the water/h-BN heterointerfaces. Utilizing this anisotropic force field, molecular dynamics simulations demonstrate that atomically flat, pristine h-BN exhibits inherent hydrophobicity. However, when atomic-step surface roughness is introduced, the wettability of h-BN undergoes a significant change, leading to a hydrophilic nature. The calculated water contact angle (WCA) for the roughened h-BN surface is approximately 64°, which closely aligns with experimental WCA values ranging from 52° to 67°. These findings indicate the high probability of the presence of atomic steps on the surfaces of the experimental h-BN samples, emphasizing the need for further experimental verification. The development of the anisotropic interfacial force field for accurately describing interactions at the water/h-BN heterointerfaces is a significant advancement in accurately simulating the wettability of two-dimensional (2D) materials, offering a reliable tool for studying the dynamic and transport properties of water at these interfaces, with implications for materials science and nanotechnology.


Prior exposure to interpersonal violence and long-term treatment response for boys with a disruptive behavior disorder.
Interpersonal violence (IPV) is common in children with a disruptive behavior disorder (DBD) and increases the risk for greater DBD symptom severity, callous-unemotional (CU) traits, and neuroendocrine disruption. Thus, IPV may make it difficult to change symptom trajectories for families receiving DBD interventions given these relationships. The current study examined whether IPV prior to receiving treatment for a DBD predicted trajectories of a variety of associated outcomes, specifically DBD symptoms, CU traits, and cortisol concentrations. Boys with a DBD diagnosis (N = 66; age range = 6-11 years; 54.5% of whom experienced IPV prior to treatment) of either oppositional defiant disorder or conduct disorder participated in a randomized clinical trial and were assessed 3 years following treatment. Multilevel modeling demonstrated that prior IPV predicted smaller rates of change in DBD symptoms, CU traits, and cortisol trajectories, indicating less benefit from intervention. The effect size magnitudes of IPV were large for each outcome (d = 0.88-1.07). These results suggest that IPV is a predictor of the long-term treatment response for boys with a DBD. Including trauma-focused components into existing DBD interventions may be worth testing to improve treatment effectiveness for boys with a prior history of IPV.


Smart, Piezo-Responsive Polyvinylidenefluoride/Polymethylmethacrylate Surface with Triggerable Water/Oil Wettability and Adhesion.
The design of smart surfaces with externally triggerable water/oil wettability and adhesion represents one of the most up-to-date challenges in the field of material science. In this work, the intelligent surface with electrically triggerable wettability and water/oil adhesion is presented. As a basic material background exhibiting electric field (EF) sensitivity, the piezo-responsive polymethylmethacrylate/polyvinylidenefluoride polymer fibers were used. To expand the available range of water/oil contact angles (CAs) and adhesion, the fibers were grafted with hydrophilic or hydrophobic functional groups using diazonium chemistry. The fiber functionality was evaluated using the static CA and wettability hysteresis measurements (increasing/decreasing drop volume and tilting angles), drops adhesion/repellence and graphite self-cleaning test performed with and without the application of EF. It was found that the proposed method enables tuning the surface wettability in the superhydrophobic/superoleophobic-hydrophilic/oleophilic range and changing of surface properties from low adhesive to high adhesive for water and oil. More convincing results were achieved in the case of fiber surface modification by ADT-C8F17, which may result from a rearrangement of the grated -C6H4C8F17 functional group under the application of EF triggering. Moreover, the triggering which can be performed in the extremely fast way (the surface responds to the EF switching on/off in seconds) was found to be fully reversible. Finally, the additional tests indicate the satisfactory stability of created fiber-based coating against the mechanical treatment.