Comparison of different analytical and precipitation methods for direct estimation of serum high-density lipoprotein cholesterol.
In this study two commercial enzymatic methods for the determination of serum cholesterol have been adapted for a continuous-flow analyser and a discrete multichannel analyser. The selective precipitation methods for lipoproteins using either dextran sulphate/MgCl2, heparin/MnCl2, phosphotungstate/MgCl2 or polyethylene glycol (mol. wt 6000) were compared with each other and with ultracentrifugal analysis as well as with agarose gel electrophoresis for lipoproteins. The results indicated that the dextran sulphate/MgCl2 precipitation procedure was the method of choice for the determination of HDL-cholesterol from clear and hyperlipemic sera. HDL-cholesterol values for 391 apparently health women (X +/- SD) were 1.58 +/- 0.34 mmol/l and for 432 men 1.27 +/- 0.29 mmol/L. The corresponding HDL-cholesterol/cholesterol ratios for women were 0.292 +/- 0.068 and for men 0.229 +/- 0.069. The HDL-cholesterol in serum was slightly age dependent in women, while the HDL-cholesterol/cholesterol ratio showed highly significant negative correlation with the age in both women and men.


[Chest granuloma secondary to methyl methacrylate. Case report].
We present the case of a patient with a history of a massive left hemithorax crushing injury in 1985; the exact management of the lesion is unknown. Twenty years later he had a thoracic fistula with a culture that was reported as positive for Enteroccocus faecalis and Staphyloccocus epidermidis. The patient was referred by the chest surgery service with the diagnosis of rib osteomyelitis once complementary imaging tests were performed (plain X-rays, CAT scan and MRI). The patient underwent surgery at our service; a granulomatous reaction secondary to a foreign body (methyl methacrylate and Ethibon) was reported. Chest reconstruction for massive lesions is possible with methyl methacrylate. Imaging studies involve the well-known difficulty to identify this material, given that it may produce signals and densities that are difficult to interpret by specialized physicians.


The Effect of a Combination of Recombinant EGF Cosmetic Serum and a Crosslinked Hyaluronic Acid Serum as Compared to a Fibroblast-Conditioned Media Serum on the Appearance of Aging Skin.
Anti-aging cosmeceutical efficacy is hampered by lack of active ingredient purity and lack of dosing standardization. These are two important key factors necessary to insure consistent, reproducible, and documentable skin effects. Without this type of standardization, it is not possible for cosmeceutical science to advance. Growth factors are interesting cosmeceutical ingredients with established cosmetic skin effects that can now be standardized due to the recent ability to manufacture recombinant epidermal growth factor. The concomitant use of a recombinant epidermal growth factor with a filler grade hyaluronic acid (EGF/RHA) was studied over 12 weeks in 60 females with mild to moderate photoaging as compared to a currently marketed spent fibroblast growth media and moisturizer (TNS). Investigator, noninvasive, and subject assessments were collected at baseline and weeks 2, 4, 8, and 12. The blinded investigator noted a statistically significant preference for the EGF/RHA at week 2 in terms of smoothness (P =0.003) and firmness (P =0.003). This improvement continued into weeks 4 and 8 with continued superior EGF/RHA results in fine lines (P =0.002), radiance (P =0.014), and overall appearance (P =0.027) by week 12. Transepidermal water loss was reduced for the EGF/RHA over the TNS at week 12 (P =0.005). The subjects gave high ratings to both study products. This research demonstrates the utility of recombinant growth factors, when combined with hyaluronic acid hydration, in improving skin cosmetic attributes. The ability to manufacture consistent pure recombinant growth factors lays the foundation for improved scientific study of this category of cosmeceutical actives.

 J Drugs Dermatol. 2016;15(6):738-741.


[Up-regulated expression of miR-576 inhibits ALK4 expression, regulates JAK/STAT signaling pathway and promotes proliferation and migration of prostatic cancer cells].
To explore the mechanism of the action of the miR-576/ALK4 axis on the progression of prostate cancer (PCa). PCa cells were transfected with miR-576 mimics/inhibitor, the proliferation and migration distance of the cells were detected by MTT and scratch wound healing assay, respectively. The targeted regulation effect of miR-576 on ALK4 was verified by dual-luciferase reporter assay. The effects of miR-576 on the mRNA and protein expressions and phosphorylation levels of the ALK4 and JAK/STAT signaling pathway factors JAK2 and STAT3 were determined by qPCR and Western blot, respectively. The C4-2 cells were co-treated with sh-ALK4 and Ruxolitinib for measurement of the proliferation and migration of the PCa cells. Bioinformatics analysis and binding site prediction showed that miR-576 was up-regulated in the PCa cells, and dual-luciferase reporter assay revealed its targeted regulation effect on ALK4 and its impact on the phosphorylation levels of JAK2 and STAT3. Overexpressed miR-576 promoted while knocked-down miR-576 inhibited the proliferation and migration of the PCa cells. sh-ALK4 increased the proliferation and migration of the cells, while Ruxolitinib suppressed the promoting effect of sh-ALK4. The expression of miR-576 is up-regulated in PCa, inhibits the expression of ALK4, regulates the activity of the JAK and STAT signaling pathways, and promotes the proliferation and migration of PCa cells.


An enzyme-linked immunosorbent assay for the quantitation of IgG anti-D and IgG subclasses in the sera of alloimmunized patients.
IgG subclass composition of maternal alloantibodies to the D antigen seems to play a role in the severity of hemolytic disease of the newborn. The subclassing of IgG anti-D is usually performed by hemagglutination techniques, but the results are not quantitative and sometimes are difficult to interpret. Thus, there is a need for quantitative methods. The aim of this study was to develop an enzyme-linked immunosorbent assay (ELISA) for the quantitation of specific IgG anti-D and IgG subclasses in the sera of alloimmunized patients. Group O R1R2 red cells were sensitized with anti-D. Red cell membranes were solubilized with nonionic detergent. IgG and IgG subclasses were measured by a sensitive and reproducible immunocapture ELISA. A serum calibrated for its IgG subclass content was used as a reference, and the anti-D preparation 68/419 was used as an internal control. Optimal conditions for the detection of IgG anti-D and IgG subclasses by ELISA were studied. The absolute concentration and the proportions of IgG subclasses were determined in the sera of 14 pregnant women. A close parallelism was observed between dilutions of the IgG reference serum and the IgG anti-D solubilized from sensitized RBCs. The sum of IgG anti-D subclass concentrations, determined by the ELISA, correlated well with other quantitative methods. The method described is sensitive and can be used routinely for the quantitative determination of specific IgG anti-D and IgG subclasses in sera.


Catastrophic visual loss due to Cryptococcus neoformans meningitis.
We have reviewed our experience with 17 of our own patients with cryptococcal meningitis and 32 cases from the literature. Although this complication is an uncommon event, patients with cryptococcal meningitis may develop visual loss in the absence of other ocular lesions (endophthalmitis or cryptococcomas in the visual pathway) that could explain the visual symptoms. There are 2 distinct patterns of visual loss: rapid visual loss and slow visual loss. Rapid visual loss is characterized by onset of profound visual loss over a period as short as 12 hours before or early in the course of therapy and a clinical syndrome that is strongly suggestive of optic neuritis. Direct invasion of the optic nerve by C. neoformans is demonstrated by cases in this and other reports. Slow visual loss is characterized by slow but progressive visual loss which typically begins later during therapy and may be due to the effects of increased intracranial pressure. While the initial deficit may be mild, patients with slow visual loss can progress to severe visual loss over weeks to months. The only factors that appear to predict either pattern of visual loss are the presence of papilledema, an elevated CSF opening pressure, and a positive CSF India ink preparation. In the 25 visual loss patients for whom data were available for all 3 items, 10 (40%) were positive for all 3, as opposed to only 4 of 114 (3.5%) from a reference group of cryptococcal meningitis patients without visual loss (p < 0.00001). The only therapeutic measures with any degree of consistent success were those directed at reducing intracranial pressure. When begun early and used aggressively, such therapy halted and sometimes even reversed the course of visual loss, particularly in the slow visual loss group. Corticosteroids did not appear to be of value in the small number of patients who received them.


A graph kernel method for DNA-binding site prediction.
Protein-DNA interactions play important roles in many biological processes. Computational methods that can accurately predict DNA-binding sites on proteins will greatly expedite research on problems involving protein-DNA interactions. This paper presents a method for predicting DNA-binding sites on protein structures. The method represents protein surface patches using labeled graphs and uses a graph kernel method to calculate the similarities between graphs. A new surface patch is predicted to be interface or non-interface patch based on its similarities to known DNA-binding patches and non-DNA-binding patches. The proposed method achieved high accuracy when tested on a representative set of 146 protein-DNA complexes using leave-one-out cross-validation. Then, the method was applied to identify DNA-binding sites on 13 unbound structures of DNA-binding proteins. In each of the unbound structure, the top 1 patch predicted by the proposed method precisely indicated the location of the DNA-binding site. Comparisons with other methods showed that the proposed method was competitive in predicting DNA-binding sites on unbound proteins. The proposed method uses graphs to encode the feature's distribution in the 3-dimensional (3D) space. Thus, compared with other vector-based methods, it has the advantage of taking into account the spatial distribution of features on the proteins. Using an efficient kernel method to compare graphs the proposed method also avoids the demanding computations required for 3D objects comparison. It provides a competitive method for predicting DNA-binding sites without requiring structure alignment.


Robotically Assisted Recipient Site Preparation in Hair Restoration Surgery: Surgical Safety and Clinical Outcomes in 31 Consecutive Patients.
Recent advances in robotic surgery have extended to hair restoration surgery, using a robotic recipient site creation device. This study aimed to assess the surgical safety and postoperative clinical outcomes of using this robotic system. Thirty-one men diagnosed with androgenetic alopecia, who underwent hair transplantation with robotic recipient site creation, were retrospectively reviewed. Their mean age was 38.7 ± 9.5 (range, 22‒67) years. The total number of robotically created recipient sites was 36,273. The average site creation speed was 1,593 ± 544 sites per hour. Postoperative crusting (54.8%) was the most frequent complication in the recipient area, followed by pruritus (12.9%), asymmetry (6.5%), and folliculitis (6.5%). The mean score of cosmetic outcomes and patient satisfaction, scored on a 5-point scale, was 4.10 ± 0.54 and 4.13 ± 0.85, respectively. No significant differences in cosmetic outcomes and patient satisfaction were found between 3 operators. The current device can automatically make slit incisions in the recipient area with speed and consistency noninferior to manual site creation. It is both safe and reliable for clinical use, and it is also easily managed by different hair surgeons without a long learning curve.


Molecular Taxonomic Profiling of Bacterial Communities in a Gilthead Seabream (Sparus aurata) Hatchery.
As wild fish stocks decline worldwide, land-based fish rearing is likely to be of increasing relevance to feeding future human generations. Little is known about the structure and role of microbial communities in fish aquaculture, particularly at larval developmental stages where the fish microbiome develops and host animals are most susceptible to disease. We employed next-generation sequencing (NGS) of 16S rRNA gene reads amplified from total community DNA to reveal the structure of bacterial communities in a gilthead seabream (Sparus aurata) larviculture system. Early- (2 days after hatching) and late-stage (34 days after hatching) fish larvae presented remarkably divergent bacterial consortia, with the genera Pseudoalteromonas, Marinomonas, Acinetobacter, and Acidocella (besides several unclassified Alphaproteobacteria) dominating the former, and Actinobacillus, Streptococcus, Massilia, Paracoccus, and Pseudomonas being prevalent in the latter. A significant reduction in rearing-water bacterial diversity was observed during the larviculture trial, characterized by higher abundance of the Cryomorphaceae family (Bacteroidetes), known to populate microniches with high organic load, in late-stage rearing water in comparison with early-stage rearing-water. Furthermore, we observed the recruitment, into host tissues, of several bacterial phylotypes-including putative pathogens as well as mutualists-that were detected at negligible densities in rearing-water or in the live feed (i.e., rotifers and artemia). These results suggest that, besides host-driven selective forces, both the live feed and the surrounding rearing environment contribute to shaping the microbiome of farmed gilthead sea-bream larvae, and that a differential establishment of host-associated bacteria takes place during larval development.


Dissecting human antibody responses: useful, basic and surprising findings.
Human memory B cells and plasma cells represent a rich source of antibodies that have been selected in response to human pathogens. In the last decade, different methods have been developed to interrogate the human memory repertoire and isolate monoclonal antibodies. I will discuss how a target-agnostic approach based on high-throughput screening of antibodies produced by cultured B cells and plasma cells has not only provided potent and broadly neutralizing antibodies against a range of pathogens, but has also advanced our understanding of basic aspects of the immune response, from host-pathogen interaction to the role of somatic mutations in affinity maturation and in the diversification of the antibody response. Most surprisingly, this approach has also revealed a new mechanism of diversification based on templated insertion of non-Ig DNA into antibody genes that we discovered in the context of the immune response to malaria infection.


Tetramethoxystilbene Inhibits NLRP3 Inflammasome Assembly via Blocking the Oligomerization of Apoptosis-Associated Speck-like Protein Containing Caspase Recruitment Domain: In Vitro and In Vivo Evaluation.
Nucleotide-binding domain leucine-rich repeat family pyrin domain containing 3 (NLRP3) inflammasome complex regulates the caspase-1 activity and subsequent processing of interleukin-1β (IL-1β). Various inflammatory diseases involve the activation of inflammasome complexes; thus, the intervention in complex formation via small molecules offers a new therapeutic opportunity. The structure-guided design and synthesis of a series of methoxystilbenes and methoxy-2-phenylnaphthalenes identified new inhibitors of NLRP3 inflammasome complex. The tetramethoxystilbene 4o and trimethoxy 2-phenylnaphthalene 1t inhibit the release of a mature form of IL-1β in J774A.1 cells with IC50 values of 1.39 and 2.07 μM, respectively. Mechanistic investigation revealed that tetramethoxystilbene 4o blocks the oligomerization of apoptosis-associated speck-like protein (ASC), which is the vital step in the formation of NLRP3 inflammasome assembly, thus preventing the activation of caspase-1 and the IL-1β release. Treatment of LPS+ATP challenged mice with 20 mg/kg of 4o significantly suppressed the levels of IL-1β. The data presented herein warrant further investigation of methoxystilbenes in disease-specific models of inflammatory diseases.


Redox regulation of the signaling pathways leading to eNOS phosphorylation.
Oxidative stress mediates positive and negative effects on physiological processes. Recent reports show that H(2)O(2) induces phosphorylation and activation of endothelial nitric oxide synthase (eNOS) through an Akt-phosphorylation-dependent pathway. In this study, we assessed activation of eNOS and Akt by determining their phosphorylation status. Whereas moderate levels of H(2)O(2) (100 microM) activated the Akt/eNOS pathway, higher levels (500 microM) did not, suggesting differential effects by differing levels of oxidative stress. We then found that two pro-oxidants with activity on sulfhydryl groups, 1-chloro-2,4-dinitrobenzene (CDNB) and diethyl maleate (DEM), blocked the phosphorylation events induced by 100 microM H(2)O(2). GSH was not a target thiol in this system because buthionine sulfoximine did not inhibit this phosphorylation. However, down-regulation of cell membrane surface and intracellular free thiols was associated with the inhibition of phosphorylation, suggesting that oxidation of non-GSH thiols inhibits the H(2)O(2)-induced phosphorylation of eNOS and Akt. DTT reversed the inhibitory effects of CDNB and DEM on Akt phosphorylation and concomitantly restored cell surface thiol levels more efficiently than it restored intracellular thiols, suggesting a more prominent role for the former. Similarly, DEM and CDNB inhibited TNF-alpha-induced Akt and eNOS phosphorylation, suggesting that thiol modification is involved in eNOS inductive pathways. Our findings suggest that eNOS activation is exquisitely sensitive to regulation by redox and that cell surface thiols, other than glutathione, regulate signal transduction leading to phosphorylation of Akt and eNOS.


Non-additive genetic effects on weights and performance of a Brazilian Bos taurus x Bos indicus beef composite.
The aim of the present study was to evaluate the heterosis effects on weaning weight at 205 days (WW, n = 146,464), yearling weight at 390 days (YW, n = 69,315) and weight gain from weaning to yearling (WG, n = 59,307) in composite beef cattle. The fixed models were: RM, which included contemporary groups, class of age of dam, outcrossing percentages for direct and maternal effects, and additive direct and maternal (AM) breed effects; R, RM model, minus AM breed effects, and H, RM model, minus additive breed effects. The estimates for W205 were in general positive (P < 0.01). The R and H models resulted in similar estimates, but they were very different from the ones estimated by the RM model. For W390, the R and H models resulted in general positive estimates (P < 0.05). For WG, the RM model resulted in general significant heterosis effects (P < 0.05). It can be concluded that the RM model seems to supply estimates of better quality (P < 0.01).


Neuroprotective effects of hypothermia.
The neuroprotective effects of hypothermia have been shown in case reports and animal studies. Therapeutic hypothermia is used to provide neuroprotection during certain types of surgery and after serious events that pose a threat to the brain. The aim of this review is to describe the efficacy of such treatment in adults. All articles retrieved from five searches in PubMed were examined. Studies were included if they had a hypothermia protocol and a measurement of neuroprotection. The list of randomised studies was completed using studies identified from five international review articles. In all, 103 of 678 studies fulfilled the inclusion criteria, of which 48 were clinical trials. Ten of the clinical trials were randomised, using a normothermic control group. Several randomised clinical trials have suggested that avoidance of hyperthermia provides the same neuroprotection as therapeutic hypothermia after cardiac arrest and traumatic brain injury, but prognostic factors and inclusion criteria vary markedly between the patient populations, including time to target temperature. Two studies found that cognitive function after prolonged aortic surgery under deep hypothermia was equivalent to that after brief normothermic interventions. Animal studies show a neuroprotective effect of hypothermia, but this is dependent on the extent of anoxic damage as well as the rate of cooling. It remains uncertain how best to implement therapeutic hypothermia to achieve neuroprotection after acute events that pose a threat to the brain. Hypothermia during aortic surgery seems to provide adequate neuroprotection for prolonged interventions.


Photophysics of the Singlet Oxygen Sensor Green Chromophore: Self-Production of 1O2 Explained by Molecular Modeling.
We report a combined computational and experimental study to rationalize the behavior of a well-known singlet oxygen (1O2) probe, that is, the chromophore of the Singlet Oxygen Sensor Green: a fluoresceine-based sensor. In particular, we evidence that the presence of an intramoleculer charge transfer state that is no more present upon reaction with 1O2 explains the fluorescence enhancement observed in the presence of reactive oxygen species. Furthermore, we also unequivocally show the photophysical pathways leading to the fluorescence enhancement of fluoresceine upon irradiation with UVA lights and also in the absence of any oxygen activator. More specifically, we evidence that the presence of a possible intersystem crossing upon population of higher energy singlet electronic excited states will lead to the population of the fluoresceine triplet manifold and hence to the self-production of 1O2.


Evaluating Injury Severity in Neonatal Encephalopathy Using Automated Quantitative Electroencephalography Analysis: A Pilot Study.
Quantitative analysis of electroencephalography (qEEG) is a potential source of biomarkers for neonatal encephalopathy (NE). However, prior studies using qEEG in NE were limited in their generalizability due to individualized techniques for calculating qEEG features or labor-intensive pre-selection of EEG data. We piloted a fully automated method using commercially available software to calculate the suppression ratio (SR), absolute delta power, and relative delta, theta, alpha, and beta power from EEG of neonates undergoing 72 h of therapeutic hypothermia (TH) for NE between April 20, 2018, and November 4, 2019. We investigated the association of qEEG with degree of encephalopathy (modified Sarnat score), severity of neuroimaging abnormalities following TH (National Institutes of Child Health and Development Neonatal Research Network [NICHD-NRN] score), and presence of seizures. Thirty out of 38 patients met inclusion criteria. A more severe modified Sarnat score was associated with higher SR during all phases of TH, lower absolute delta power during all phases except rewarming, and lower relative delta power during the last 24 h of TH. In 21 patients with neuroimaging data, a worse NICHD-NRN score was associated with higher SR, lower absolute delta power, and higher relative beta power during all phases. QEEG features were not significantly associated with the presence of seizures after correction for multiple comparisons. Our results are consistent with those of prior studies using qEEG in NE and support automated qEEG analysis as an accessible, generalizable method for generating biomarkers of NE and response to TH. Additionally, we found evidence of an immature relative frequency composition in neonates with more severe brain injury, suggesting that automated qEEG analysis may have a use in the assessment of brain maturity.


Caffeine intake and low birth weight: a population-based case-control study.
The authors conducted a matched case-control study to investigate the effects of caffeine intake during pregnancy on birth weight. From January to November 1992, in the first 24 hours after delivery, 1,205 mothers (401 cases and 804 controls) were interviewed and their newborns were examined to assess birth weight and gestational age by means of the method of Capurro et al. (J Pediatr 1978;93:120-2). The cases were children with birth weight < 2,500 g and gestational age > or = 28 weeks. Cases and controls were matched for time of birth and hospital of delivery and were recruited from the four maternity hospitals in Pelotas, southern Brazil. Daily maternal caffeine intake during pregnancy for each trimester was estimated. To assess caffeine intake, 10% of the mothers were reinterviewed at their households and samples of reported information on drip coffee and maté (a caffeine-containing drink widely used in South America) were collected and sent to the laboratory for caffeine determination through liquid chromatography. When instant coffee was reported, the weight of powder was measured using a portable scale, and caffeine intake was estimated from a reference table. Caffeine intake from tea, chocolate, soft drinks, and medicines was estimated from a reference table. Analyses were performed by conditional logistic regression. Crude analyses showed no effect of caffeine on low birth weight, preterm births or intrauterine growth retardation. The results did not change after allowing for confounders.


EBARDenovo: highly accurate de novo assembly of RNA-Seq with efficient chimera-detection.
High-accuracy de novo assembly of the short sequencing reads from RNA-Seq technology is very challenging. We introduce a de novo assembly algorithm, EBARDenovo, which stands for Extension, Bridging And Repeat-sensing Denovo. This algorithm uses an efficient chimera-detection function to abrogate the effect of aberrant chimeric reads in RNA-Seq data. EBARDenovo resolves the complications of RNA-Seq assembly arising from sequencing errors, repetitive sequences and aberrant chimeric amplicons. In a series of assembly experiments, our algorithm is the most accurate among the examined programs, including de Bruijn graph assemblers, Trinity and Oases. EBARDenovo is available at http://ebardenovo.sourceforge.net/. This software package (with patent pending) is free of charge for academic use only. Supplementary data are available at Bioinformatics online.


How to define variation of physical properties normal to an undulating one-dimensional object.
One-dimensional flexible objects are abundant in physics, from polymers to vortex lines to defect lines and many more. These objects structure their environment and it is natural to assume that the influence these objects exert on their environment depends on the distance from the line object. But how should this be defined? We argue here that there is an intrinsic length scale along the undulating line that is a measure of its stiffness (i.e., orientational persistence), which yields a natural way of defining the variation of physical properties normal to the undulating line. We exemplify how this normal variation can be determined from a computer simulation for the case of a so-called bottle-brush polymer, where side chains are grafted onto a flexible backbone.


Development and testing of hierarchically wrinkled coatings for marine antifouling.
We report on the formation and testing of novel marine coatings comprising hierarchically wrinkled surface topographies (HWTS) having wrinkles of different length scales (generations) ranging from tens of nanometers to a fraction of a millimeter. The individual wrinkle generations are arranged in nested patterns, where each larger wrinkle resides underneath and represents a scaled-up version of the smaller wrinkle. We present and discuss results from field tests in seawater and laboratory experiments. The results of our field tests reveal that while coatings with flat topographies foul after relatively short time periods (4-15 weeks), the HWST coatings with the same chemistries as flat coatings remain relatively free of biofouling even after prolonged exposure to seawater (18 months). In contrast to flat coatings, the HWST substrates are not colonized by barnacles. These observations suggest that surface topography plays a dominant role in governing the coating defense against barnacle fouling even without fine-tuning the chemical composition of the overcoat. Laboratory experiments indicate that settlement of zoospores of the green alga Ulva and the strength of attachment of sporelings (young plants) depend on the chemical composition of the coating as well as surface topography.


Occurrence and Behavior of Macrolide Antibiotics in Municipal Wastewater Treatment: Possible Importance of Metabolites, Synthesis Byproducts, and Transformation Products.
A one-year study on the occurrence and fate of macrolide antibiotics and their metabolites, synthesis byproducts, and transformation products (TPs) was performed in the wastewater treatment plant of the city of Zagreb (Croatia). The target compounds were found in all analyzed influent and effluent samples with the total concentrations of azithromycin-, clarithromycin-, and erythromycin-related compounds reaching up to 25, 12, and 0.25 μg/L, respectively. The most prominent individual constituents were the parent macrolides azithromycin and clarithromycin. However, a substantial contribution of their derivatives, formed by deglycolysation and microbial phosphorylation, was also detected. In addition, widespread presence of several linearized nontarget TPs was confirmed for the first time in real wastewater samples by suspect screening analysis. Complex characterization of macrolide-derived compounds enabled decoupling of industrial and therapeutic sources from the in situ transformations. Due to the high inputs and incomplete removal and/or formation of several TPs during the conventional wastewater treatment, the average mass load of azithromycin-related compounds in secondary effluents exceeded 3.0 g/day/1000 inhabitants. This is the first study to reveal the importance of metabolites, byproducts, and TPs for the overall mass balance of macrolide antibiotics in urban wastewater systems.


Fifteen-minute consultation: the child with systemic arterial hypertension.
Hypertension in children is a relatively uncommon condition but its prevalence is increasing with the rising rates of excess weight during childhood. It is important that hypertension is diagnosed early in children, particularly in those with symptoms, in those with secondary causes and in those with target organ damage. This article presents a systematic approach to the evaluation of a child with arterial hypertension, highlighting important points on history and examination, out-of-office monitoring and baseline investigations before consideration for more detailed investigations and treatment.


Advanced multifocal tuberculous spondylitis without disk involvement and with multidrug-resistant bacilli.
In this brief report, we describe a 23-year-old immunocompetent Nigerian patient with extensive multifocal tuberculous spondylitis without disk involvement. Cultures of sputum samples and biopsy samples from the L4 vertebra were positive for tuberculosis; drug-susceptibility testing of the isolates revealed multidrug resistance. Treatment with second-line drugs resulted in an excellent interim outcome after 6 months, without the need for surgical intervention.


Influence of bilateral cochlear implants on vocal control.
Receiving a cochlear implant (CI) can improve fundamental frequency (F0) control for deaf individuals, resulting in increased vocal pitch control. However, it is unclear whether using bilateral CIs, which often result in mismatched pitch perception between ears, will counter this benefit. To investigate this, 23 bilateral CI users were asked to produce a sustained vocalization using one CI, the other CI, both CIs, or neither. Additionally, a set of eight normal hearing participants completed the sustained vocalization task as a control group. The results indicated that F0 control is worse with both CIs compared to using the ear that yields the lowest vocal variability. The results also indicated that there was a large range of F0 variability even for the relatively stable portion of the vocalization, spanning from 6 to 46 cents. These results suggest that bilateral CIs can detrimentally affect vocal control.


Deferoxamine Counteracts Cisplatin Resistance in A549 Lung Adenocarcinoma Cells by Increasing Vulnerability to Glutamine Deprivation-Induced Cell Death.
Glutamine, like glucose, is a major nutrient consumed by cancer cells, yet these cells undergo glutamine starvation in the cores of tumors, forcing them to evolve adaptive metabolic responses. Pharmacologically targeting glutamine metabolism or withdrawal has been exploited for therapeutic purposes, but does not always induce cancer cell death. The mechanism by which cancer cells adapt to resist glutamine starvation in cisplatin-resistant non-small-cell lung cancer (NSCLC) also remains uncertain. Here, we report the potential metabolic vulnerabilities of A549/DDP (drug-resistant human lung adenocarcinoma cell lines) cells, which were more easily killed by the iron chelator deferoxamine (DFO) during glutamine deprivation than their parental cisplatin-sensitive A549 cells. We demonstrate that phenotype resistance to cisplatin is accompanied by adaptive responses during glutamine deprivation partly via higher levels of autophagic activity and apoptosis resistance characteristics. Moreover, this adaptation could be explained by sustained glucose instead of glutamine-dominant complex II-dependent oxidative phosphorylation (OXPHOS). Further investigation revealed that cisplatin-resistant cells sustain OXPHOS partly via iron metabolism reprogramming during glutamine deprivation. This reprogramming might be responsible for mitochondrial iron-sulfur [Fe-S] cluster biogenesis, which has become an "Achilles' heel," rendering cancer cells vulnerable to DFO-induced autophagic cell death and apoptosis through c-Jun N-terminal kinase (JNK) signaling. Finally, in vivo studies using xenograft mouse models also confirmed the growth-slowing effect of DFO. In summary, we have elucidated the adaptive responses of cisplatin-resistant NSCLC cells, which balanced stability and plasticity to overcome metabolic reprogramming and permitted them to survive under stress induced by chemotherapy or glutamine starvation. In addition, for the first time, we show that suppressing the growth of cisplatin-resistant NSCLC cells via iron chelator-induced autophagic cell death and apoptosis was possible with DFO treatment. These findings provide a solid basis for targeting mitochondria iron metabolism in cisplatin-resistant NSCLC for therapeutic purposes, and it is plausible to consider that DFO facilitates in the improvement of treatment responses in cisplatin-resistant NSCLC patients.


Treatment of an amelogenesis imperfecta with restorations prepared using a modified clear matrix technique.
The rehabilitation of a patient with amelogenesis imperfecta (AI) from both functional and esthetic standpoints represents a challenge. A number of treatment options have been proposed. Recently, the use of adhesive restorations has gained popularity because of the improved physical properties of these materials. This article describes a treatment with direct resin composite for the restoration of teeth affected by the hypomature type of AI. A modified clear matrix technique was used during the preparation and restoration process. The use of the technique provides clinicians with reduced chair time, and the matrix can be repeated when needed; restorations can be performed relatively quickly with a minimal post-operative finishing process. This article highlights the use of direct-bonded resin composites providing satisfactory esthetics and function in restoring AI-affected teeth.


"Assault-related shame mediates the association between negative social reactions to disclosure of sexual assault and psychological distress": Correction to Decou et al. (2017).
Reports an error in "Assault-related shame mediates the association between negative social reactions to disclosure of sexual assault and psychological distress" by Christopher R. DeCou, Trevor T. Cole, Shannon M. Lynch, Maria M. Wong and Kathleen C. Matthews (Psychological Trauma: Theory, Research, Practice, and Policy, 2017[Mar], Vol 9[2], 166-172). In the article, there was an error in the coding of missing values thus effecting the abstract, Methods, Results, and Discussion sections. The frequency counts for sexual assault victimization, reactions to social disclosure, and assault-related shame were calculated incorrectly due to an error in the coding of missing values, and have been corrected in the description of participants and in the results and discussion sections. In addition, the sample size was incorrectly reported as N = 207, and should have appeared as "N = 208." The sample size and corresponding percentages have been corrected throughout the text. Two transcription errors for the indirect effects via PTSD and global distress were also corrected. These indirect effects were incorrectly reported as "PCL-C; β = .27," and "OQ-45.2;β = .21," and should have appeared as "PCL-C;β = .26," and "OQ-45.2; β = .20." (The following abstract of the original article appeared in record 2016-43136-001.) Objective: Several studies have identified associations between social reactions to disclosure of sexual assault and psychological distress; however, no studies have evaluated shame as a mediator of this association. This study evaluated assault-related shame as a mediator of the associations between negative social reactions to disclosure of sexual assault and symptoms of posttraumatic stress disorder (PTSD), depression, and global distress and hypothesized that there would be an indirect effect of social reactions to disclosure upon symptoms of psychopathology via assault-related shame. Participants were 207 female psychology undergraduates who reported past history of completed or attempted sexual assault and had disclosed the assault to at least 1 other person. Participants completed self-report measures of social reactions to sexual assault disclosure, assault-related shame, and symptoms of psychopathology. Participants reported significant histories of attempted or completed sexual assault and indicated clinically significant symptoms of depression and subthreshold symptoms of PTSD and global distress, on average. Evaluation of structural models confirmed the hypothesized indirect effect of negative social reactions to sexual assault disclosure upon symptoms of PTSD (z = 5.85, p < .001), depression (z = 4.56, p < .001), and global distress (z = 4.82, p < .001) via assault-related shame. These findings offer new insight concerning the intervening role of assault-related shame and highlight the importance of shame as a target for therapeutic intervention. This study suggests the need for future research concerning the role of shame in the etiology of PTSD and process of disclosure among survivors of attempted or completed sexual assault. (PsycINFO Database Record


[Application of plasmapheresis in complex of preoperative preparation in patients with autoimmune thyroiditis and diffuse toxic goiter].
The article is devoted to actulal problem of endocrinology and surgery-treatment of severe diseases of the thyroid gland. Results of plasmapheresis application in complex preoperative preparation in 90 patients, aged from 20 to 60 yrs, in severe course of the disease, were adduced. After plasmapheresis preparation conduction the reduction of the autoimmune processes in the patients organisms, the antibodies titer lowering were noted, witnessing the stable remission achievement. Plasmapheresis application in complex of treatment of patients with autoimmune thyroiditis and diffuse toxic goiter causes significant positive influence on hormonal and immune state.


The association between psychosis and violent crime: a study of offenders evaluated at a court psychiatric clinic.
Examination of 2100 psychiatric evaluations conducted by a court psychiatric clinic to determine competency or for a presentence evaluation indicated no association between a diagnosis of psychosis and commission of a crime of violence. However, offenders referred for a pretrial competency examination were more likely to have committed a crime of violence than were defendants seen for a presentence evaluation. The implications of these results are discussed.


Hyperzincemia in anencephaly and spina bifida: a clue to the pathogenesis of neural tube defects?
Zinc is essential for normal embryogenesis and may have particular importance for closure of the human neural tube. Compared to 258 controls, we found increased zinc content in umbilical cord serum in eight of nine newborn anencephalics (p less than 0.02) and three infants with spina bifida (p less than 0.001). Increased zinc levels were bound to serum albumin or alpha-2-macroglobulin (alpha 2M) in infants with neural tube defects (NTD). In NTD-mothers, total serum zinc was similar to controls, but there was a shift in the distribution of zinc from alpha 2M to albumin. Fetal hyperzincemia and elevated maternal albumin-bound zinc suggest that the NTD-fetus receives but does not use zinc normally.


Scanning electron microscopy of the lungs of vitamin E-deficient rats exposed to a low concentration of ozone.
The acute effects of a low concentration of ozone on the surface morphologic features of the conducting airways and alveoli were studied by scannging electron microscopy. Rats were exposed to approximately 0.3 ppm of ozone for 3 hours daily for as many as 16 consecutive days. Animals were killed at intervals of time, and their lungs were processed. Throughout the airways, cilia were seen to be swollen, showing hemispheric extrusions and considerable surface roughness. Groups of severely injured cilia had a tendency to adhere to each other. Small, round bodies were frequently noted, mainly in the large airways and the proximal bronchioles, in some areas thickly covering the airway epithelial surface. In addition, the luminal surfaces of the epithelium of the regions were often covered with a pseudomembrane. The surfaces of protruding nonciliated cells (Clara cells) showed many small swellings and discrete, round bodies. These changes were most marked in the terminal airways. The surfaces of alveolar ducts and alveolar walls showed scattered areas of cytoplasmic swelling and attachment of round bodies throughout the parenchyma. The findings described were more prominent in vitamin E-deficient rats than in rats fed a diet supplemented with vitamin E, suggesting an antioxidant effect of the vitamin.


Effect of Water Content on Enthalpic Relaxations in Porcine Septal Cartilage.
Cartilage thermoforming is an emerging surgical technology which uses heat to accelerate stress relaxation in mechanically deformed tissue specimens. Heat induced shape change in cartilage is associated with complex thermo mechanical behavior of which the mechanisms are still a subject of debate. Differential scanning calorimetry (DSC) was used to characterize the threshold temperatures and enthalpies in cartilage as a function of water content. The DSC identified two enthalpic events in porcine nasal septal cartilage, which depend on the water content. The change in the water content of cartilage impacts the interactions between matrix macromolecules and water molecules, which may be associated with a bound-free water transformation (reversible process) and a denaturation of cartilage (irreversible process).


Predictive Value of RAPID Assessed Perfusion Thresholds on Final Infarct Volume in SWIFT PRIME (Solitaire With the Intention for Thrombectomy as Primary Endovascular Treatment).
Computed tomography perfusion imaging can estimate the size of the ischemic core, which can be used for the selection of patients for endovascular therapy. The relative cerebral blood volume (rCBV) and relative cerebral blood flow (rCBF) thresholds chosen to identify ischemic core influence the accuracy of prediction. We aimed to analyze the accuracy of various rCBV and rCBF thresholds for predicting the 27-hour infarct volume using RAPID automated analysis software from the SWIFT PRIME trial (Solitaire With the Intention for Thrombectomy as Primary Endovascular Treatment) data. Patients from the SWIFT PRIME study who achieved complete reperfusion based on time until the residue function reached its peak >6 s perfusion maps obtained at 27 hours were included. Patients from both the intravenous tissue-type plasminogen activator only and endovascular groups were included in analysis. Final infarct volume was determined on magnetic resonance imaging (fluid-attenuated inversion recovery images) or computed tomography scans obtained 27 hours after symptom onset. The predicted ischemic core volumes on rCBV and rCBF maps using thresholds ranging between 0.2 and 0.8 were compared with the actual infarct volume to determine the most accurate thresholds. Among the 47 subjects, the following baseline computed tomography perfusion thresholds most accurately predicted the actual 27-hour infarct volume: rCBV=0.32, median absolute error (MAE)=9 mL; rCBV=0.34, MAE=9 mL; rCBF=0.30, MAE=8.8 mL; rCBF=0.32, MAE=7 mL; and rCBF=0.34, MAE=7.3. Brain regions with rCBF 0.30 to 0.34 or rCBV 0.32 to 0.34 thresholds provided the most accurate prediction of infarct volume in patients who achieved complete reperfusion with MAEs of ≤9 mL. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01657461.


Histological and Histomorphometrical Analysis on a Loaded Implant With Platform-Switching and Conical Connection: A Case Report.
The association of Morse taper implant-abutment design with the use of a smaller abutment diameter (platform switching) may improve dental implant success rate and prevent peri-implant bone loss. The aim of the present study was to histologically and histomorphometrically evaluate the behavior of peri-implant tissues around an implant with a conical connection associated with platform switching. A platform-switched Morse-cone connection implant was inserted in the left posterior mandible of a 61-year-old patient. The implant was inserted at the level of the alveolar crest. After 11 months from placement and 6 months of loading, the implant was retrieved for psychological reasons and processed for histological evaluation. The retrieved implant was wholly surrounded by bone tissue, except for a small area in the apical portion. At higher magnification, in the coronal portion of the implant, it was possible to observe bone directly at the implant platform level. No resorption of the coronal bone was present, except for 0.2 mm on the vestibular aspect. Crestally, bone remodeling with areas of newly formed bone was detected; the bone-implant contact was 73.9%. Apposition of bone was detected even upon the platform. Peri-implant crestal bone preservation can be achieved with the combination of Morse taper conical internal implant-abutment connection with the use of a smaller abutment diameter (platform-switching).


GENETICS OF PHYSIOLOGICAL AND BEHAVIORAL RESISTANCE TO HOST FURANOCOUMARINS IN THE PARSNIP WEBWORM.
Depressaria pastinacella, the parsnip webworm, feeds almost exclusively on the flowers and fruits of Pastinaca sativa, the wild parsnip. Resistance to webworms in wild parsnip populations is largely attributable to genetically based variation in furanocoumarin chemistry; by differentially reducing fruit set among chemical phenotypes, parsnip webworms may act as selective agents on wild parsnip populations. To determine whether wild parsnip chemistry can act as a selective agent on webworm populations, it is necessary to establish that resistance mechanisms in the webworm to furanocoumarins are genetically based. In this study, we estimated the amount of genetic variation in behavioral and physiological responses of webworms to parsnip furanocoumarins. Virtually no variation was found among webworm families for feeding preferences for diets varying as much as fourfold in furanocoumarin content. Nor was significant variation found for mean furanocoumarin intake over the assay period, except in one case, in which maternal effects may account for differences among families. In contrast, substantial familial variation existed for cytochrome P450-mediated metabolism of bergapten and xanthotoxin, two host furanocoumarins. The presence of additive genetic variation in metabolism, and the absence of such variation in discriminative feeding behavior, suggests that adaptation to changes in furanocoumarin chemistry, resulting either from changes in the distribution of chemical phenotypes in parsnip populations or from shifts to new chemically different host plants, is likely to be facilitated by physiological rather than behavioral means.


Super-Expansion of Assembled Reduced Graphene Oxide Interlayers by Segregation of Al Nanoparticle Pillars for High-Capacity Na-Ion Battery Anodes.
The applicability of Na-ion batteries is contingent on breakthroughs in alternative electrode materials that have high capacities and which are economically viable. Unfortunately, conventional graphite anodes for Li-ion battery systems do not allow Na-ion accommodation into their interlayer space owing to the large ionic radius and low stabilizing energy of Na in graphite. Here, we suggest a promising strategy for significantly increasing Na capacity by expanding the axial slab space of graphite. We successfully synthesized reconstructed graphite materials via self-assembly of negative graphite oxide (GO) flakes and Al cation (positive) pillars and by subsequent chemical reaction of the obtained Al-GO materials. Al pillars, atomically distributed in graphite interlayers, can extend the slab space by up to ∼7 Å, which is a 2-fold interlayer distance of pristine graphite. An exceptionally high capacity of 780 mAh/g is demonstrated for reconstructed graphite anodes with Al pillars, compared with rGO materials (210 mAh/g). We investigated the electrochemical reaction mechanism and structural changes associated with discharge and charge to emphasize the benefit of using reconstructed graphite as anodes in Na-ion batteries. Our strategy of modifying the interlayer distance by introducing metallic pillars between the layers can help address the low capacity of carbonaceous anodes.


Lymphatic delivery of etanercept via nanotopography improves response to collagen-induced arthritis.
Evidence suggests lymphatic function mediates local rheumatoid arthritis (RA) flares. Yet biologics that target the immune system are dosed systemically via the subcutaneous (SC) administration route, thereby inefficiently reaching local lymphatic compartments. Nanotopography has previously been shown to disrupt tight cellular junctions, potentially enhancing local lymphatic delivery and potentially improving overall therapeutic efficacy. We first characterized nanotopography (SOFUSA™) delivery of an anti-TNF drug, etanercept, by comparing pharmacokinetic profiles to those obtained by conventional SC, intravenous (IV), and intradermal (ID) routes of administration, and assessed uptake of radiolabeled etanercept in draining lymph nodes (LNs) in single dosing studies. We then compared etanercept efficacy in a progressive rat model of collagen-induced arthritis (CIA), administered systemically via SC route of administration; via the regional lymphatics through ID delivery; or through a nanotopography (SOFUSA™) device at 10, 12, and 14 days post CIA induction. Measurements of hind limb swelling and near-infrared fluorescence (NIRF) imaging of afferent lymph pumping function and reflux were conducted on days 11, 13, and 18 post CIA induction and compared to untreated CIA animals. Univariate and multivariate analysis of variance were used to compare the group differences for percentage swelling and lymphatic contractile activity. Even though all three modes of administration delivered an equal amount of etanercept, SOFUSA™ delivery resulted in increased lymphatic pumping and significantly reduced swelling as compared to untreated, ID, and SC groups. Pharmacokinetic profiles in serum and LN uptake studies showed that using the nanotopography device resulted in the greatest uptake and retention in draining LNs. Locoregional lymphatic delivery of biologics that target the immune system may have more favorable pharmacodynamics than SC or IV administration. Nanotopography may provide a more efficient method for delivery of anti-TNF drugs to reverse impairment of lymphatic function and reduce swelling associated with RA flares.


Genetic alterations in locally advanced stage II/III colon cancer: a search for prognostic markers.
Heterogeneity in advanced colon cancer leads to different results from adjuvant chemotherapy. To identify groups of patients who may need adjuvant treatment, molecular staging and correlation with clinical data may be helpful in classifying histologically similar tumors. Colon cancer develops through a multistep process with an accumulation of multiple genetic alterations that are often the cause of a form of genomic instability. The 2 best known mechanisms of genomic instability are chromosomal instability (CIN) and microsatellite instability (MSI). The CIN phenotype is found in approximately 85% of sporadic colon cancers and is characterized by aneuploidy, multiple chromosomal rearrangements, and an accumulation of somatic mutations in oncogenes such as K-ras and tumor suppressor genes such as TP53 and APC. The MSI phenotype is associated with small insertions and deletions mainly in repetitive sequences (microsatellites) and is found in approximately 15% of cases. This instability, often referred to as high-frequency MSI (MSI-H), is caused by defects of the mismatch repair system, which is involved in repairing DNA errors that arise during DNA replication. Clear-cut correlations between the somatic genetic alterations in tumors and the clinical behavior of the tumor are rare. Only a few markers, such as MSI-H and TP53, seem to have a prognostic value. Mutations in the TP53 gene are associated with an aggressive tumor growth and subsequent reduced survival, whereas MSI-H seems to be correlated with a favorable outcome. In general, predicting biologic behavior of in particular stage III colon cancers is difficult and remains a great clinical problem.


New fatherhood: myths and realities.
Despite evidence for the existence of nurturing fathers, little is known about how and why men increase their involvement with their children. The qualitative study from which these findings were taken explored the experience of 10 Canadian fathers in dual-earner families after the birth of their first infants. The constant comparative method was used to generate a grounded theory explicating the men's process of redefining their roles as spouses, workers, and fathers. Findings from 20 semi-structured interviews conducted with a purposive sample of fathers in their homes revealed that the fathers' experiences consisted of coping with many demands from children, partners, and jobs. These men redefined their roles as fathers after their partners returned to full-time employment. Two stages of monitoring role strain and limiting role strain captured the variation in behavior. This study contributes to community health nurses' understanding of how men respond to the lack of concurrence among societal expectations, others' expectations, and their own realities of fatherhood.


VEGF gene delivery for treatment of ischemic cardiovascular disease.
There are increasing numbers of patients with ischemic myocardial disease not amenable to traditional methods of revascularization. These patients may benefit from new research into the use of naturally occurring angiogenic compounds, such as vascular endothelial growth factor (VEGF) for re-establishing blood flow into regions of hibernating myocardium. Animal studies and human clinical trials evaluating VEGF demonstrate increases in myocardial perfusion after treatment, with some patients reporting improvement in anginal symptoms. Further research into the ideal form of VEGF therapy (protein, plasmid, or adenoviral) and delivery method (intracoronary, intramyocardial, or epicardial) seems justified.


Characterization of the T-cell epitopes of the major peach allergen Pru p 3.
Pru p 3 is the major peach allergen recognized by more than 90% of peach-allergic individuals of the Mediterranean area. Identification of the dominant Pru p 3 T-cell epitopes can improve our understanding of the immune responses against this protein and could be helpful in the development of hypoallergenic immunotherapy. For this purpose, we examined the phenotypes, specificities and cytokine secretion profiles of proliferating T cells in response to Pru p 3 in peach-allergic individuals. Peripheral blood mononuclear cells from 15 peach-allergic patients were incubated with Pru p 3. The proliferation of antigen-specific T-cell lines (TCLs) was assessed by tritiated methylthymidine incorporation. T-cell epitopes were identified by analyzing the reactivity of TCLs against 8 overlapping peptides spanning the entire length of Pru p 3. We characterized the phenotype of Pru-p-3-specific TCLs by flow cytometry and analyzed their production of interleukin (IL) 4 and gamma-interferon (IFN-gamma) by ELISA. Ninety-two Pru-p-3-specific TCLs were isolated (stimulation index > or =5). These TCLs proliferated mainly in response to Pru p 3(12-27) and Pru p 3(57-72). Pru-p-3-specific TCLs were mainly CD4+ (81%) and expressed cell surface CD30. In addition, TCLs produced high levels of IL-4 and low levels of IFN-gamma, indicating a Th2 phenotype. Two immunodominant T-cell-reactive regions of Pru p 3 were identified: Pru p 3(12-27) and Pru p 3(57-72). These peptides showed a differential ability to elicit a Th2 response. Taken together, our results provide a better understanding of the immunological T-cell reactivity against Pru p 3.


[Curative effect of surgery on moyamoya disease: an analysis of 47 patients].
To evaluate the effect of surgery on 47 patients with moyamoya disease by retrospective analysis. Methods: A total of 47 patients with moyamoya disease were enrolled from August, 2010 to According to the improved treatment in August, 2013, all cases were divided into two groups: a pre-improved group and a post-improved group. According to different surgical methods, they were divided into two subgroups: an indirect revascularization subgroup and a combined revascularization subgroup. Results: The cerebral ischemia in 77.4% of patients was relieved after the surgery. There was significant difference in outcomes of patients between the pre-improved group and the post-improved group (P<0.05), while there was no significant difference between the pre-improved indirect revascularization subgroup and the pre-improved combined revascularization subgroup. There was also no significant difference between the post-improved indirect revascularization subgroup and the post-improved combined revascularization subgroups (P>0.05). Conclusion: Surgical treatment can improve the outcomes of patients with moyamoya disease, but there is no significant difference in surgical effects between indirect and combined revascularization.


[Neuropsychologic contribution to dyslexia research. A review of important explanatory models and findings].
Since in a substantial proportion of reading-disabled children constitutional factors appear to play an important pathogenetic role, major contributions to a better understanding of dyslexia have long come from the fields of neuropsychology and behavioral neurology. In this overview an attempt is made to describe the most important hypotheses and models on which neuropsychological research on this problem has been based. The first section is devoted to a discussion of findings on acquisition of reading and spelling skills by brain-damaged children and on the effect of the localization of central nervous system lesions on reading disorders in children. Classic neurological disorders are seen in only a small proportion of children with reading retardation, however. Therefore the second section comprises a discussion of various hypotheses that have been developed to explain the particular performance profiles seen in children with dyslexia but with no evidence of brain damage, i.e. in those children who can be said to have a developmental disorder. It is emphasized that dyslexia is probably not a uniform syndrome but that rather different subgroups of reading-disabled children can be identified.


Metabolic responses and nutritional therapy in patients with severe head injuries.
The severe hypermetabolism and hypercatabolism seen in patients with severe head injuries results in malnutrition that occurs very rapidly and can cause impaired healing and an increased tendency to infection and multiple organ failure. Thus, early adequate nutritional support plays a role in functional outcome. Total enteral nutrition (TEN) is preferred over total parenteral nutrition (TPN), but TPN should be supplied promptly while increasing TEN to a goal of at least 25 to 35 nonprotein kcal/kg/d and 2.0 to 2.5 g protein/kg/d. Nutritional formulas high in branched chain amino acids, glutamine, arginine, vitamins E and C, and zinc may also have some advantages. Growth hormone may improve anabolism. Hyperglycemia, especially glucose levels exceeding 200 mg/dL, must be prevented and/or treated promptly with insulin or decreased glucose intake. Careful monitoring with indirect calorimetry and nitrogen balance studies should help prevent inadequate protein or excessive carbohydrate intake.


The tissue factor requirement in blood coagulation.
Formation of thrombin is triggered when membrane-localized tissue factor (TF) is exposed to blood. In closed models of this process, thrombin formation displays an initiation phase (low rates of thrombin production cause platelet activation and fibrinogen clotting), a propagation phase (>95% of thrombin production occurs), and a termination phase (prothrombin activation ceases and free thrombin is inactivated). A current controversy centers on whether the TF stimulus requires supplementation from a circulating pool of blood TF to sustain an adequate procoagulant response. We have evaluated the requirement for TF during the progress of the blood coagulation reaction and have extended these analyses to assess the requirement for TF during resupply ("flow replacement"). Elimination of TF activity at various times during the initiation phase indicated: a period of absolute dependence (<10 s); a transitional period in which the dependence on TF is partial and decreases as the reaction proceeds (10-240 s); and a period in which the progress of the reaction is TF independent (>240 s). Resupply of reactions late during the termination phase with fresh reactants, but no TF, yielded immediate bursts of thrombin formation similar in magnitude to the original propagation phases. Our data show that independence from the initial TF stimulus is achieved by the onset of the propagation phase and that the ensemble of coagulation products and intermediates that yield this TF independence maintain their prothrombin activating potential for considerable time. These observations support the hypothesis that the transient, localized expression of TF is sufficient to sustain a TF-independent procoagulant response as long as flow persists.


Childhood cutaneous hemangiomas.
An hemangioma is a common benign growth of vascular endothelium that may have multiple clinical manifestations. Either identified at birth or shortly thereafter, hemangiomas have both proliferative and involutional phases. Certain varieties are associated with congenital anomalies. Numerous therapy options are discussed.


From 2D framework to quasi-1D nanomaterial: preparation, characterization, and formation mechanism of Cu(3)SnS(4) nanorods.
An ion-ion reaction route under solvothermal condition at relatively low temperatures was first put forward to the preparation of tetragonal Cu(3)SnS(4) nanorods, on the basis of the strategy that 2D framework structure of Cu(3)SnS(4) containing layers could provide orientation for the growth of quasi-1D nanomaterials. X-ray diffraction (XRD) pattern, transmission electronic microscope (TEM) images, electronic diffraction (ED) pattern, X-ray photoelectron spectra (XPS), energy-dispersive X-ray analysis (EDXA), Mössbauer spectrum, Raman spectrum, thermal analysis (DTA and TGA), ultraviolet and visible light (UV-vis) spectrum, and photoluminescence (PL) spectrum were used to characterize the products. On the basis of a series of supplementary experiments and the result of infrared absorption spectrum (IR), a reaction mechanism was proposed: ethanol as solvent and reductant and trace water/CH(3)CSNH(2) as sulfur source and acid-making components could form 2D network through hydrogen bonds, which provided the orientation for the formation of a 2D framework structure; appropriate concentration of CH(3)CSNH(2), warming speed, reaction constant temperatures (T(rc)), and reaction time also played important roles.


Generation of Marked and Markerless Mutants in Model Cyanobacterial Species.
Cyanobacteria are ecologically important organisms and potential platforms for production of biofuels and useful industrial products. Genetic manipulation of cyanobacteria, especially model organisms such as Synechocystis sp. PCC6803 and Synechococcus sp. PCC7002, is a key tool for both basic and applied research. Generation of unmarked mutants, whereby chromosomal alterations are introduced into a strain via insertion of an antibiotic resistance cassette (a manipulatable fragment of DNA containing one or more genes), followed by subsequent removal of this cassette using a negative selectable marker, is a particularly powerful technique. Unmarked mutants can be repeatedly genetically manipulated, allowing as many alterations to be introduced into a strain as desired. In addition, the absence of genes encoding antibiotic resistance proteins in the mutated strain is desirable, as it avoids the possibility of 'escape' of antibiotic resistant organisms into the environment. However, detailed methods for repeated rounds of genetic manipulation of cyanobacteria are not well described in the scientific literature. Here we provide a comprehensive description of this technique, which we have successfully used to generate mutants with multiple deletions, single point mutations within a gene of interest and insertion of novel gene cassettes.


The effect of training and epinephrine infusion on ratings of perceived exertion (RPE).
Seven untrained males (mean age [+/-SD] = 25.6+/-3.9 yr, mean ht = 177.0+/-5.9 cm, mean wt = 65.8+/-7.4 kg) completed a 6-week exercise program (cycle ergometer). Prior to training, and at the end of each week of training, each subject performed a 20 min constant-power exercise test (absolute power was the same each week). At the end of the six week training program (within a few days), an additional 20 min constant-power test was performed, during which epinephrine was infused at a rate of 100 ng x kg(-1) x min(-1) over the final 10 min of exercise. Training significantly (P<0.05) reduced end-exercise ratings of perceived exertion (RPE), plasma epinephrine concentration [Epi], plasma norepinephrine concentration [NE], blood lactate concentration [La-], minute ventilation (V(E)), heart rate (HR), and blood glucose concentration [Glc]. Epinephrine infusion failed to increase RPE despite significant (p < 0.05) increases in [Epi], [La-], V(E) and [Glc]. Therefore, the present data indicate that RPE during exercise is not causally related to changes in plasma [Epi]. It also appears that modest changes in plasma [NE], blood [La-], V(E) and blood [Glc] during constant-power cycle ergometry (as observed during Epi infusion) do not impact RPE.


The nutritional needs of the older person.
Older people have particular nutritional requirements that are different from other groups. The nutritional status of many older people is poor in respect of some key nutrients. Older people are at higher risk of malnutrition if they are ill, live alone or have difficulty in eating. Older patients should have their nutritional status and nutritional intake monitored regularly to ensure they are receiving adequate nutrition.


Accounting for outliers and heteroskedasticity in multibreed genetic evaluations of postweaning gain of Nelore-Hereford cattle.
The objectives of this study were to demonstrate the utility of hierarchical Bayesian models combining residual heteroskedasticity with robustness for outlier detection and muting and to evaluate the effects of such joint modeling in multibreed genetic evaluations. A 3 x 2 factorial specification of 6 residual variance models based on several distributional (Gaussian, Student's t, or Slash) and variability (homoskedastic or heteroskedastic) assumptions was used to analyze 22,717 postweaning gain records from a Nelore-Hereford population (40,082 animals in the pedigree). To illustrate the utility of the 2 robust distributional specifications (Student's t and Slash) for outlier detection and muting, 3 records from the same contemporary group (an extreme residual outlier, a mild residual outlier, and a near-zero residual) were chosen for further study. The posterior densities of the corresponding weighting variables of these records were used to assess their degree of Gaussian outlyingness and the ability of the robust models to mute the effects of deviant records. The Student's t heteroskedastic provided the best-fit model among the 6 specifications and was preferred for genetic merit inference. Kendall rank correlations of the posterior means of the additive genetic effects of the animals, used to compare the selection order of the Student's t and Gaussian models, were reasonably high across all animals within the most frequent genotypes, ranging from 0.83 to 0.91 and from 0.89 to 0.95 for the homoskedastic and the heteroskedastic versions, respectively. However, when considering only animals ranked in the top 10% by the customary Gaussian homoskedastic model, these rank correlations were reduced considerably, ranging from 0.29 to 0.57 and from 0.72 to 0.85 between the 2 residual densities within the homoskedastic and heteroskedastic versions, respectively. Rank correlations between the homoskedastic and heteroskedastic versions within each of the Gaussian and Student's t error models tended to be smaller, with a range from 0.68 to 0.90 across all animals and from 0.28 to 0.67 for animals ranked in the top 10%. These results support the implementation of robust models accounting for sources of heteroskedasticity to increase the precision and stability of multibreed genetic evaluations with proper statistical treatment of deviant records.


Agreement and reproducibility in identification of endometriosis using magnetic resonance imaging.
Magnetic resonance imaging (MRI) is a promising technique in the study of endometriosis, allowing a complete mapping of lesions before surgery. However, the value of MRI in the diagnosis of endometriosis in the bladder, in superficial peritoneal lesions, and in ovarian foci and uterosacral ligaments is still under debate. To assess inter- and intra-observer agreement in the evaluation of endometriosis in different anatomical locations using MRI. From June 2006 to February 2008, 83 female patients (mean age 39, range 19-49 years) who had undergone MRI examination for suspected endometriosis were evaluated by two radiologists. MRI at 1.5 Tesla was performed with SE and TSE sequences, T1- and T2-weighted with and without fat suppression. Each examination was completed with gadolinium administration. Each dataset was independently evaluated by the radiologists for the presence or absence of endometriosis. The location (ovaries, uterosacral ligaments (USLs), pouch of Douglas, vagina, rectosigmoid, rectovaginal septum, and bladder) of suspected lesions was recorded. Cohen kappa statistical analysis was performed to calculate agreement between measurements. After 2 months the data were analyzed again by the two observers to assess intra-observer agreement. Of the 83 MRI examinations performed, 12 patients demonstrated no evidence of endometriosis. In the remaining 71 studies, 157 endometriotic lesions ranging in size from 0.4 to 6.2 cm were detected. Of the 157 lesions, 53 (33.75% incidence) were smaller than 1 cm. In the ovaries, the inter-observer agreement was 92.77% and the kappa value was 0.802 (95% CI, 0.695-0.91). In the bladder the inter-observer agreement was 96.39% and the kappa value was 0.553 (95% CI 0.056-1). In the USLs the inter-observer agreement was 90.96% and the kappa value 0.583 (95% CI, 0.381-0.784). In the rectovaginal septum the inter-observer agreement was 94.58% and the kappa value 0.739 (95% CI, 0.572-0.905). In the rectovaginal pouch the inter-observer agreement was 88.55% and the kappa value 0.608 (95% CI, 0.443-0.774). In the vaginal fornix the inter-observer agreement was 94.58% and the kappa value 0.726 (95% CI, 0.552-0.901). In the rectosigmoid the inter-observer agreement was 89.76% and the kappa value 0.589 (95% CI, 0.389-0.768). The results of our study indicate that MRI has a high inter- and intra-observer agreement in the identification of endometriosis located in the ovary, rectosigmoid, and rectovaginal septum, whereas the agreement is suboptimal for the identification of endometriosis located in the USLs.


Investigating DNA adduct formation by flavor chemicals and tobacco byproducts in electronic nicotine delivery system (ENDS) using in silico approaches.
The presence of flavors is one of the commonly cited reasons for use of e-cigarettes by youth; however, the potential harms from inhaling these chemicals and byproducts have not been extensively studied. One mechanism of interest is DNA adduct formation, which may lead to carcinogenesis. We identified two chemical classes of flavors found in tobacco products and byproducts, alkenylbenzenes and aldehydes, documented to form DNA adducts. Using in silico toxicology approaches, we identified structural analogs to these chemicals without DNA adduct information. We conducted a structural similarity analysis and also generated in silico model predictions of these chemicals for genotoxicity, mutagenicity, carcinogenicity, and skin sensitization. The empirical and in silico data were compared, and we identified strengths and limitations of these models. Good concordance (80-100%) was observed between DNA adduct formation and models predicting mammalian mutagenicity (mouse lymphoma sassy L5178Y) and skin sensitization for both chemical classes. On the other hand, different prediction profiles were observed for the two chemical classes for the modeled endpoints, unscheduled DNA synthesis and bacterial mutagenicity. These results are likely due to the different mode of action between the two chemical classes, as aldehydes are direct acting agents, while alkenylbenzenes require bioactivation to form electrophilic intermediates, which form DNA adducts. The results of this study suggest that an in silico prediction for the mouse lymphoma assay L5178Y, may serve as a surrogate endpoint to help predict DNA adduct formation for chemicals found in tobacco products such as flavors and byproducts.


Devising a Method to Study if Wernicke's Aphasia Patients are Aware That They Do Not Comprehend Language or Speak It Understandably.
Wernicke's Aphasia (WA) is characterized by an individual speaking fluent gibberish without the ability to understand anything that is said to them or anything they attempt to read. It is caused by damage to the left posterior temporoparietal cortex, also known as Wernicke's area. An additional intriguing symptom of WA patients is their apparent obliviousness to their own lack of understanding despite their intact reasoning or other cognitive abilities. Their only deficit seems to be in the basic rules of language that define word meaning, also known as phonology. Growing out of a project in an undergraduate class, we devised a phonology-free approach to communicating with WA patients that attempts to answer the questions of whether WA patients know that they do not understand what is said to them, that others do not understand what they have said, and if these patients are distressed by this lack of communication. We here describe the process and the resulting method.


Case report: A toxoplasmic encephalitis in an immunocompromised child detected through metagenomic next-generation sequencing.
There exist numerous pathogens that are capable of causing infections within the central nervous system (CNS); however, conventional detection and analysis methods prove to be challenging. Clinical diagnosis of CNS infections often depends on clinical characteristics, cerebrospinal fluid (CSF) analysis, imaging, and molecular detection assays. Unfortunately, these methods can be both insensitive and time consuming, which can lead to missed diagnoses and catastrophic outcomes, especially in the case of infrequent diseases. Despite the application of appropriate prophylactic regimens and evidence-based antimicrobial agents, CNS infections continue to result in significant morbidity and mortality in hospital settings. Metagenomic next-generation sequencing (mNGS) is a novel tool that enables the identification of thousands of pathogens in a target-independent manner in a single run. The role of this innovative detection method in clinical pathogen diagnostics has matured over time. In this particular research, clinicians employed mNGS to investigate a suspected CNS infection in a child with leukemia, and unexpectedly detected Toxoplasma gondii. A 3-year-old child diagnosed with T-cell lymphoblastic lymphoma was admitted to our hospital due to a 2-day history of fever and headache, along with 1 day of altered consciousness. Upon admission, the patient's Glasgow Coma Scale score was 14. Brain magnetic resonance imaging revealed multiple abnormal signals. Due to the patient's atypical clinical symptoms and laboratory test results, determining the etiology and treatment plan was difficulty.Subsequently, the patient underwent next-generation sequencing examination of cerebrospinal fluid. The following day, the results indicated the presence of Toxoplasma gondii. The patient received treatment with a combination of sulfamethoxazole (SMZ) and azithromycin. After approximately 7 days, the patient's symptoms significantly improved, and they were discharged from the hospital with oral medication to continue at home. A follow-up polymerase chain reaction (PCR) testing after about 6 weeks revealed the absence of Toxoplasma. This case highlights the potential of mNGS as an effective method for detecting toxoplasmic encephalitis (TE). Since mNGS can identify thousands of pathogens in a single run, it may be a promising detection method for investigating the causative pathogens of central nervous system infections with atypical features.


Screening for atypical porcine pestivirus in Swedish boar semen used for artificial insemination and a characterisation of the seminal RNA microbiome including the virome.
This study aimed to characterise the RNA microbiome, including the virome of extended semen from Swedish breeding boars, with particular focus on Atypical porcine pestivirus (APPV). This neurotropic virus, associated with congenital tremor type A-II in piglets, was recently demonstrated to induce the disease through insemination with semen from infected boars. From 124 Artificial Insemination (AI) doses from Swedish breeding boars, APPV was detected in one dose in addition to a sparse seminal RNA virome, characterised by retroviruses, phages, and some fecal-associated contaminants. The detected seminal microbiome was large and characterized by Gram-negative bacteria from the phylum Proteobacteria, mainly consisting of apathogenic or opportunistic bacteria. The proportion of bacteria with a pathogenic potential was low, and no antimicrobial resistance genes (ARGs) were detected in the datasets. Overall, the results indicate a good health status among Swedish breeding boars. The detection of APPV in semen raises the question of whether routine screening for APPV in breeding boars should be instigated.


Importance of the alpha 3-fragment of complement C4 for the binding with C4b-binding protein.
The human regulatory complement component C4b-binding protein (C4BP) is a multimeric plasma protein, which regulates the classical pathway of the complement system. C4BP functions as a cofactor to factor 1 in the degradation of C4b and accelerates the decay rate of the C4b2a complex. Previously, we have demonstrated that monoclonal antibodies (C4-2 and 9) directed against the alpha'-chain of C4b inhibit the binding of C4b to C4BP. In order to identify the structural domain of C4b that binds C4BP, proteolytic fragments of C4 were generated with trypsin and Staphylococcus aureus V8 protease. Sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting and amino acid sequence analysis of the proteolytic fragments reactive with the anti-C4 mAb's revealed that the residues Ala738-Arg826 of the alpha 3-fragment of C4b are important for the interaction with C4BP.


Synthesis of pH-responsive N-acetyl-cysteine modified starch derivatives for oral delivery.
In this study, a novel type of pH-responsive polymer PyHES-NAC (2-hydroxy-3-(2-propynyloxy) propyl hydroxyethyl starch (PyHES)) - (N-acetyl-cysteine (NAC)) was synthesized. First, PyHES was prepared via hydrophobic modification of hydroxyl groups in hydroxyethyl starch (HES) with propynylglycidyl ether (PGE), and then pH-responsive carboxylic acid group was connected to propynyl group via thiol-yne click reaction with NAC. Aqueous PyHES-NAC solutions exhibited a good transference between hydrophobic (or self-assembly) and hydrophilic static along with the change of pH value and protective properties of drugs under acidic conditions. 10.0% DOX was released under artificial gastric fluid after 2 h, whereas an immediate release (above 80%) was observed under artificial intestinal fluid. Drug loading capacity of PyHES-NAC was increased by the increase of degree of substitution (DS) of hydrophobic propynyl groups in PyHES, and 41 wt% DOX Loading capacity was the highest value in our study area.


Using body donor demographics to assist the implementation of donation programs in Brazil.
The use of human material in anatomy education depends upon the generosity of body donors. However, little is known regarding the demographics of body donors in Brazil, where voluntary body donation is a relatively rare phenomenon. Hence, the aim of the present study was to elucidate the demographic profile of applicants to the Body Donation Program (BDP) at the Federal University of Health Sciences of Porto Alegre in Brazil, as well as to assess whether the observed characteristics of body donors are unique to that sample, or if they merely reflect the characteristics of the regional population. Information derived from the specific forms filled out by donors between January 2008 and June 2016 at the time of registration were collected. Data from 416 forms were analyzed. Based on this study, the typical applicant in Brazil is typically a white female (67.4%), over 60 years of age (60.3%), unmarried or single (70.6%), affiliated with a religious group (89.1%), of middle class background (40.4%), who has completed high school and/or holds a university degree (93.8%). The motivation of donors was, in most cases, an altruistic gesture, represented by the desire to help society and science. Elucidating these demographic characteristics of potential donors may help identify the target public to which information regarding body donation campaigns could be directed. Anat Sci Educ 10: 475-486. © 2017 American Association of Anatomists.


Mental steps: Differential activation of internal pacemakers in motor imagery and in mental imitation of gait.
Gait imagery and gait observation can boost the recovery of locomotion dysfunctions; yet, a neurologically justified rationale for their clinical application is lacking as much as a direct comparison of their neural correlates. Using functional magnetic resonance imaging, we measured the neural correlates of explicit motor imagery of gait during observation of in-motion videos shot in a park with a steady cam (Virtual Walking task). In a 2 × 2 factorial design, we assessed the modulatory effect of gait observation and of foot movement execution on the neural correlates of the Virtual Walking task: in half of the trials, the participants were asked to mentally imitate a human model shown while walking along the same route (mental imitation condition); moreover, for half of all the trials, the participants also performed rhythmic ankle dorsiflexion as a proxy for stepping movements. We found that, beyond the areas associated with the execution of lower limb movements (the paracentral lobule, the supplementary motor area, and the cerebellum), gait imagery also recruited dorsal premotor and posterior parietal areas known to contribute to the adaptation of walking patterns to environmental cues. When compared with mental imitation, motor imagery recruited a more extensive network, including a brainstem area compatible with the human mesencephalic locomotor region (MLR). Reduced activation of the MLR in mental imitation indicates that this more visually guided task poses less demand on subcortical structures crucial for internally generated gait patterns. This finding may explain why patients with subcortical degeneration benefit from rehabilitation protocols based on gait observation. Hum Brain Mapp 38:5195-5216, 2017. © 2017 Wiley Periodicals, Inc.


[Prevalence and significance of coronary collateral circulation in patients with acute myocardial infarct].
Angiograms from consecutive and unselected patients with acute myocardial infarction were studied with respect to the prevalence as well as the significance of coronary collateral circulation to myocardium distal to the acute coronary occlusion. Coronary angiograms were obtained from 700 consecutive and unselected patients with an acute transmural infarction within 3.7 +/- 3 hours (0.5-12) of symptom onset. No patient had undergone i.v. thrombolysis prior to angiography. Complete and acute vessel occlusion was found in 626/700 patients (89%). Coronary collaterals were detected and graded using Rentrop's classification. The grade of collateral circulation was related to the clinical course after 30 days and to the global and regional left ventricular wall motion. Collaterals were found in 334 patients (69%); 242 patients (38%) had collateral flow grade 2 or 3. Collaterals were demonstrated more frequently in women vs men and in patients with multivessel disease. The prevalence of collaterals was unrelated to age and the presence of diabetes mellitus. Patients who had angiography within 3 hours of symptom onset had collaterals detected less frequently than patients who had angiography beyond 6 hours (66% vs 75%, p < 0.05). No collaterals were found in 17/37 patients (47%) in cardiogenic shock and inferior MI but in only 30/164 patients (18%, p < 0.01) without shock. Global and regional left ventricular wall motion after 2 weeks was unrelated to the degree of coronary collateral circulation during acute myocardial infarction. Collateral circulation to myocardium distal to an acutely occluded coronary artery is detected in 2/3 patients during the acute infarct phase. The absence of collaterals is related to the early occurrence of cardiogenic shock in patients with inferior MI but not to the presence of diabetes mellitus. After direct angioplasty of the infarct vessel, the protective effects of coronary collaterals on chronic LV function remain uncertain.


A novel piezoelectric biosensor for the detection of phytohormone beta-indole acetic acid.
A novel piezoelectric immunosensor has been developed for the determination of beta-indole acetic acid (IAA) in dilute solutions. The detection is based on competitive immunoreaction between a hapten (IAA) and an antigen (IAA-BSA, hapten-protein conjugation) bound to an anti-IAA antibody, immobilized on a quartz crystal microbalance (QCM). The frequency change (y) of the sensor caused by antigen is linearly related to the logarithm of the concentration of IAA (x) in the range of 0.5 ng/ml - 5 microg/ml with a regression equation of the form y = -23x + 151 (r = 0.9937).


Role of protein modifications mediated by transglutaminase 2 in human viral diseases.
Transglutaminase 2 (TG2) belongs to a family of calcium-dependent enzymes that catalyze transamidation reaction, producing polymerized, polyaminated or deamidated proteins. Recently, a growing number of viral proteins as well as cellular proteins with which they interact have been found to be modified by TG2, suggesting a novel function for TG2 in viral pathogenesis. This review summarizes the results of relevant research, examines the mechanisms underlying TG2 function in host-virus interactions and proposes a model for viral pathogenesis involving TG2.


Fold-stratified cross-validation for unbiased and privacy-preserving federated learning.
We introduce fold-stratified cross-validation, a validation methodology that is compatible with privacy-preserving federated learning and that prevents data leakage caused by duplicates of electronic health records (EHRs). Fold-stratified cross-validation complements cross-validation with an initial stratification of EHRs in folds containing patients with similar characteristics, thus ensuring that duplicates of a record are jointly present either in training or in validation folds. Monte Carlo simulations are performed to investigate the properties of fold-stratified cross-validation in the case of a model data analysis using both synthetic data and MIMIC-III (Medical Information Mart for Intensive Care-III) medical records. In situations in which duplicated EHRs could induce overoptimistic estimations of accuracy, applying fold-stratified cross-validation prevented this bias, while not requiring full deduplication. However, a pessimistic bias might appear if the covariate used for the stratification was strongly associated with the outcome. Although fold-stratified cross-validation presents low computational overhead, to be efficient it requires the preliminary identification of a covariate that is both shared by duplicated records and weakly associated with the outcome. When available, the hash of a personal identifier or a patient's date of birth provides such a covariate. On the contrary, pseudonymization interferes with fold-stratified cross-validation, as it may break the equality of the stratifying covariate among duplicates. Fold-stratified cross-validation is an easy-to-implement methodology that prevents data leakage when a model is trained on distributed EHRs that contain duplicates, while preserving privacy.


Cutaneous reactive angiomatosis associated with chronic lymphoid leukemia.
Cutaneous reactive angiomatosis is an unusual benign vascular disorder of the skin usually associated to systemic diseases. It is characterized by lobular or diffuse proliferation of small blood vessels with hyperplasia of endothelial cells, pericytes, and sometimes histiocytes. We report a 59-year-old man with asymptomatic erythematous-violaceous patches on back, palms, and elbows for 9 months. Laboratory examination revealed changes consistent with B-chronic lymphocytic leukemia. Cutaneous biopsy showed a predominantly lobular small blood vessel proliferation in dermis with pericytic hyperplasia and mild perivascular lymphoplasmacytic infiltrate. Spontaneous involution of lesions occurred after 6 months. A second biopsy performed at the beginning of clinical involution showed a less prominent vascular component with perivascular giant cells with coexpression of CD68 and CD 31. To our knowledge, this is the first case of cutaneous reactive angiomatosis with documented histopathological findings of clinical involution.


Frequent upregulation of HER2 protein in hormone-receptor-positive HER2-negative breast cancer after short-term neoadjuvant endocrine therapy.
Endocrine resistant metastatic disease develops in ~ 20-25% of hormone-receptor-positive (HR+) breast cancer (BC) patients despite endocrine therapy (ET) use. Upregulation of HER family receptor tyrosine kinases (RTKs) represent escape mechanisms in response to ET in some HR+ tumors. Short-term neoadjuvant ET (NET) offers the opportunity to identify early endocrine escape mechanisms initiated in individual tumors. This was a single arm, interventional phase II clinical trial evaluating 4 weeks (± 1 week) of NET in patients with early-stage HR+/HER2-negative (HER2-) BC. The primary objective was to assess NET-induced changes in HER1-4 proteins by immunohistochemistry (IHC) score. Protein upregulation was defined as an increase of ≥ 1 in IHC score following NET. Thirty-seven patients with cT1-T3, cN0, HR+/HER2- BC were enrolled. In 35 patients with evaluable tumor HER protein after NET, HER2 was upregulated in 48.6% (17/35; p = 0.025), with HER2-positive status (IHC 3+ or FISH-amplified) detected in three patients at surgery, who were recommended adjuvant trastuzumab-based therapy. Downregulation of HER3 and/or HER4 protein was detected in 54.2% of tumors, whereas HER1 protein remained low and unchanged in all cases. While no significant volumetric reduction was detected radiographically after short-term NET, significant reduction in tumor proliferation rates were observed. No significant associations were identified between any clinicopathologic covariates and changes in HER1-4 protein expression on multivariable analysis. Short-term NET frequently and preferentially upregulates HER2 over other HER family RTKs in early-stage HR+/HER2- BC and may be a promising strategy to identify tumors that utilize HER2 as an early endocrine escape pathway. Trial registration number: NCT03219476.


An alternative process for cleaning knives used on meat slaughter floors.
Traditionally on slaughter floors operator knives are cleaned by rinsing in hand wash water at 20-40 degrees C followed by brief immersion in baths termed "sterilisers" which contain water no cooler than 82 degrees C. Under Australian legislation, both domestic and export, it is possible for a meat processing establishment to apply to the Controlling Authority for permission to implement an alternative procedure providing that it is at least the equivalent of that legislated. No firm evidence appears to exist for the 82 degrees C requirement and the possibility of replacing this element of the knife cleaning procedure with an alternative procedure using 60 degrees C water and a longer immersion time was investigated at an abattoir slaughtering cattle and sheep. Knives were tested at a range of work stations located along beef and mutton slaughter floors for Aerobic Plate Counts (APCs) and E. coli. For knives used on the beef chain the mean log APC/cm(2) was 2.18 by the current knife cleaning process and 1.78 by the alternate procedure (P<0.001). Using the current system E. coli was isolated from cleaned knives on 20/230 (8.7%) occasions compared with 21/230 (9.1%) occasions using the alternative system. The mean log E. coli of positive knives was 0.43/cm(2) and 0.61/cm(2) from the current and alternative systems, respectively. On the mutton chain the mean log APC/cm(2) was 1.95 using the current knife cleaning process and 1.69 by the alternative procedure (P=0.014). Using the current system E. coli was isolated from cleaned knives on 24/130 (18.5%) occasions compared with 29/130 (22.3%) occasions using the alternative system. The mean log E. coli of positive knives was 0.90/cm(2) and 0.76/cm(2) from the current and alternative systems, respectively. It is concluded that using two knives alternatively, rinsing them in hand wash water, then immersing them between uses in 60 degrees C water provides a microbiological outcome equivalent to rinsing them and momentary dipping in 82 degrees C water.


Maintenance treatment in metastatic breast cancer.
Metastatic breast cancer (MBC) occurs in 20-30% of women with breast cancer and is an incurable disease. Treatment is palliative and directed to prolong survival, decrease symptoms and improve patients' quality of life. For patients with hormone receptor-negative disease or for hormone receptor-positive disease that has become resistant to endocrine therapy, or is progressing rapidly and life threatening, cytotoxic chemotherapy is indicated. However, the optimal duration of chemotherapy treatment for MBC is still a matter of debate. Studies using maintenance chemotherapy regimens standard in the 1990s showed a consistent benefit with a more prolonged time to progression, although an improvement in survival was only demonstrated in one study. Two recent trials with newer cytotoxic agents showed controversial results; whereas one study concluded that the policy of prolonging treatment in chemotherapy-sensitive patients, after aggressive, modern combination chemotherapy, cannot be recommended for women with MBC, the other study showed that maintenance therapy with pegylated liposomal doxorubicin significantly prolonged time to progression in MBC patients after first-line chemotherapy without significant clinical toxicity. Initial data regarding metronomic chemotherapy indicate that continuously low-dose cyclophosphamide and methotrexate is minimally toxic and effective in heavily pretreated breast cancer patients. In daily practice, maintenance chemotherapy is a reasonable strategy that prolongs time to progression in patients with MBC who did not show progression after first-line chemotherapy. However, this benefit should be considered together with toxicities of treatment and the patient's preference.