Twenty-Four Genes are Upregulated in Patients with Hypospadias.
Hypospadias is a congenital hypoplasia of the penis, with displacement of the urethral opening along the ventral surface, and has been reported to be one of the most common congenital anomalies, occurring in approximately 1:250 to 1:300 live births. As hypospadias is reported to be an easily diagnosed malformation at the crossroads of genetics and environment, it is important to study the genetic component in order to elucidate its etiology. In this study, the gene expression profiles both in human hypospadias tissues and normal penile tissues were studied by Human Gene Expression Array. Twenty-four genes were found to be upregulated. Among these, ATF3 and CYR61 have been reported previously. Other genes that have not been previously reported were also found to be upregulated: BTG2, CD69, CD9, DUSP1, EGR1, EIF4A1, FOS, FOSB, HBEGF, HNRNPUL1, IER2, JUN, JUNB, KLF2, NR4A1, NR4A2, PTGS2, RGS1, RTN4, SLC25A25, SOCS3 and ZFP36 (p <0.05). Further studies including genome-wide association studies (GWAS) with expression studies in a large patient group will help us for identifiying the candidate gene(s) in the etiology of hypospadias.


T-cell factor-1 expression during human natural killer cell development and in circulating CD56(+) bright natural killer cells.
Transcription factors are essential to govern differentiation along the lymphoid lineage from uncommitted hematopoietic stem cells. Although many of these transcription factors have putative roles based on murine knockout experiments, their function in human lymphoid development is less known and was studied further. Transcription factor expression in fresh and cultured adult human bone marrow and umbilical cord blood progenitors was evaluated. We found that fresh CD34(+)Lin(-) cells that are human leukocyte antigen (HLA)-DR(-) or CD38(-) constitutively express GATA-3 but not T-cell factor-1 (TCF-1) or Id-3. Culture with the murine fetal liver cell line AFT024 and defined cytokines was capable of inducing TCF-1 mRNA. However, no T-cell receptor gene rearrangement was identified in cultured progeny. Id-3, a basic helix loop helix factor with dominant negative function for T-cell differentiation transcription factors, also was upregulated and may explain unsuccessful T-cell maturation. To better understand the developmental link between natural killer (NK) cells derived from progenitors, we studied NK cell subsets circulating in blood. CD56(+bright), but not CD56(+dim), NK cells constitutively express TCF-1 by reverse transcriptase polymerase chain reaction and Western blot analysis. The TCF-1 isoform found in CD56(+bright) cells, which express lectin but not immunoglobulin class I recognizing inhibitory receptors, was identical to that induced in NK cell differentiation culture and was distinctly different from isoforms in T cells. These results suggest that TCF-1 does not target human killer immunoglobulin receptor genes, TCF-1 is uniquely expressed in circulating CD56(+bright) NK cells, and specific TCF-1 isoforms may play an important role in regulating NK differentiation from a common NK/T-cell progenitor.


Steric strain versus hyperconjugative stabilization in ethane congeners.
Rotation barriers in the group IVB ethane congeners H(3)X-YH(3) (X, Y = C, Si, Ge, Sn, Pb) have been systematically studied and deciphered using the ab initio valence bond theory in terms of the steric strain and hyperconjugation effect. Our results show that in all cases the rotation barriers are dominated by the steric repulsion whereas the hyperconjugative interaction between the X-H bond orbitals and the vicinal Y-H antibond orbitals (and vice versa) plays a secondary role, although indeed the hyperconjugation effect favors staggered structures. By the independent estimations of the hyperconjugative and steric interactions in the process of rotations, we found that the structural effect which mainly refers to the central X-Y bond relaxation makes a small contribution to the rotational barriers. Therefore, we conclude that both the rigid and fully relaxed rotations in the group IVB ethane congeners H(3)X-YH(3) observe the same mechanism which is governed by the conventional steric repulsion.


Paracrine regulation of lung maturation.
Although there is much yet to be learned about the regulation of fetal lung maturation, the study of this process has yielded much information that may serve as a paradigm for the understanding of fetal growth regulation in general. Two particularly interesting fetal adjustments of general regulatory phenomena have emerged: the ability of local tissue metabolism to anatomically or temporally isolate endocrine effects and the role of paracrine factors in coordinating the responses of different cells in complex organs.


Liquid-liquid miscibility gaps in drug-water binary systems: crystal structure and thermodynamic properties of prilocaine and the temperature-composition phase diagram of the prilocaine-water system.
EMLA cream, a "eutectic mixture of local anesthetics", was developed in the early 1980s by Astra Pharmaceutical Production. The mixture of anesthetics containing lidocaine, prilocaine, and water is liquid at room temperature, which is partly due to the eutectic equilibrium between prilocaine and lidocaine at 293 K, as was clear from the start. However, the full thermodynamic background for the stability of the liquid and its emulsion-like appearance has never been elucidated. In the present study of the binary system prilocaine-water, a region of liquid-liquid demixing has been observed, linked to a monotectic equilibrium at 302.4 K. It results in a prilocaine-rich liquid containing approximately 0.7 mol fraction of anesthetic. Similar behavior has been reported for the binary system lidocaine-water (Céolin, R.; et al. J. Pharm. Sci. 2010, 99 (6), 2756-2765). In the ternary mixture, the combination of the monotectic equilibrium and the above-mentioned eutectic equilibrium between prilocaine and lidocaine results in an anesthetic-rich liquid that remains stable below room temperature. This liquid forms an emulsion-like mixture in the presence of an aqueous solution saturated with anesthetics. Physical properties and the crystal structure of prilocaine are also reported.


Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. IV. The less frequently expressed VL are heterogeneous.
We previously demonstrated that the human anti-Haemophilus influenzae type b polysaccharide (Hib-PS) VL repertoire is dominated by a product of the V kappa II gene, A2, and that V kappa II-A2 anti-Hib-PS antibodies have little or no somatic mutation in VL. To further study this VL repertoire, we studied non-A2 anti-Hib-PS antibodies that were identified either serologically or by amino-terminal amino acid sequence analysis. Of 15 non-A2 anti-Hib-PS antibodies from 12 vaccinated adults, we found four V lambda, five V kappa I, one non-A2 V kappa II, four V kappa III, and one V kappa IV antibodies. As expected, all but two of these subjects also produced V kappa II-A2 antibodies. Interestingly, one of these subjects lacks the A2 gene in the germ line. However, both subjects who did not produce detectable V kappa II antibody did produce normal amounts of total anti-Hib-PS antibody after vaccination. Candidate V kappa genes for the non-A2 antibodies were identified by comparison of up to 60 VL amino acid residues, including CDR1 and CDR2, with all sequenced V kappa genes. V kappa I antibodies appear to be products of three newly sequenced V kappa I genes, O8, O18, and L11, that are reported here. The O8 and O18 genes encode identical amino acid sequences. The non-A2 V kappa II antibody is a likely product of the A1 or A17 genes, the V kappa III antibodies are likely products of the A27 gene, and the V kappa IV antibody is a product of the single V kappa IV gene, B3. Unlike V kappa II-A2 antibodies, the V kappa I, V kappa III, and V kappa IV antibodies differed by one to five CDR residues from the germ line product of the candidate genes, suggesting the presence of somatic mutations. Thus, anti-Hib-PS antibodies can be divided into two types, the most frequently observed A2 antibodies with little or no somatic mutation and non-A2 antibodies that likely contain somatic mutations.


Lymph node examination. What is an adequate workup?
Lymph node examination has traditionally been difficult to perform satisfactorily. Both pathologists and clinicians often lack confidence in the resulting diagnosis, and repeated biopsy or routine requests for consultative opinion may follow. Artifactual distortion of lymph nodes during removal and processing are partially responsible for these unsatisfactory circumstances. More recently, a developing awareness of the inherent complexity of lymphoproliferative diseases and the availability of more sophisticated studies have further compounded these traditional problems. This article describes the specific methods recommended to allow community pathologists to scale lymph node examination to the diagnostic difficulty of the disease present. In particular, simple and inexpensive procedures allow tissue samples to be saved for immunologic phenotyping and electron microscopy while routine histopathologic sections are being evaluated.


Intermittent self-catheterisation.
Intermittent self-catheterisation (ISC) has reduced infection hazards and greatly improved the lives of many patients with micturition disorders. The technique is used by male and female patients who experience neurogenic bladder dysfunction or voiding difficulties. Some of the most beneficial results associated with ISC have been reported when the technique is used by people with spina bifida, paraplegia or multiple sclerosis. ISC is the preferred course of management for patients with a hypotonic bladder (a floppy bladder that is unable to contract partially or totally during the micturition cycle).


Engineering an E. coli Near-Infrared Light Sensor.
Optogenetics is a technology wherein researchers combine light and genetically engineered photoreceptors to control biological processes with unrivaled precision. Near-infrared (NIR) wavelengths (>700 nm) are desirable optogenetic inputs due to their low phototoxicity and spectral isolation from most photoproteins. The bacteriophytochrome photoreceptor 1 (BphP1), found in several purple photosynthetic bacteria, senses NIR light and activates transcription of photosystem promoters by binding to and inhibiting the transcriptional repressor PpsR2. Here, we examine the response of a library of output promoters to increasing levels of Rhodopseudomonas palustris PpsR2 expression, and we identify that of Bradyrhizobium sp. BTAi1 crtE as the most strongly repressed in Escherichia coli. Next, we optimize Rps. palustris bphP1 and ppsR2 expression in a strain engineered to produce the required chromophore biliverdin IXα in order to demonstrate NIR-activated transcription. Unlike a previously engineered bacterial NIR photoreceptor, our system does not require production of a second messenger, and it exhibits rapid response dynamics. It is also the most red-shifted bacterial optogenetic tool yet reported by approximately 50 nm. Accordingly, our BphP1-PpsR2 system has numerous applications in bacterial optogenetics.


Environmental impact of desalination technologies: A review.
Due to the limited availability of freshwater supplies, desalination has become an increasingly reliable process for water supply worldwide, with proved technical and economic feasibility and advantages. Recently, desalination capacity significantly increased from approximately 35 million m3 daily (MCM/day) in 2005 to about 95 MCM/day in 2018. Seawater desalination accounts for about 61% of global desalination capacity, while brackish water desalination accounts for 30%. Membrane desalination, mainly using reverse osmosis (RO), accounts for ¾ of global desalination capacity, with the rest mostly used for thermal desalination using multi-stage flash distillation (MSF), and multi-effect distillation (MED). Despite the undeniable role of desalination for securing water supply in areas where natural freshwater supplies are scarce, desalination impacts the natural environment at different aspects. Environmental impacts (EIs) of the desalination process are different and vary significantly according to the nature of the utilized feedwater, the desalination technology in use, and the management of waste brine generated. In this work, the EIs of each desalination technology were thoroughly investigated, with careful consideration given to different feedwater qualities, and various brine management techniques. Although the different aspects of desalination EIs have been extensively studied in the literature, the literature lacks comprehensive reviews and summaries of all the associated EIs. This article compiles the different EIs associated with the whole desalination process in one-hub, applying an intake-to-outfall approach. The leading desalination technologies of RO, MSF, and MED were analyzed, along with different feedwaters. This article provides a mapping of the different technologies involving feedwater and brine management techniques and a detailed description of their impact on the environment. Finally, recommendations and conclusions were given to minimize the negative impacts of desalination on both the local and global environments.


Effects of Molecular Weight and Concentration of Poly(Acrylic Acid) on Biomimetic Mineralization of Collagen.
Inspired by nature, poly(acrylic acid) (PAA) and other polyelectrolytes have been used as noncollagenous proteins (NCPs) surrogates for biomimetic intrafibrillar mineralization of collagen fibrils and thus, to model the ultrastructure of bone, to study the mechanism of bone mineralization and, more scarcely to fabricate scaffolds for hard tissue engineering. The objective of this study was to systematically investigate the effect of the molecular weight (MW) and the concentration of PAA on the rate and pattern of biomineralization of collagen matrices. Densified type I collagen films were mineralized in supersaturated PAA-stabilized amorphous calcium-phosphate (PAA-ACP) solutions containing increasing MW (2 kDa, 50 kDA, 450 kDa) and concentrations (10, 25, 50 mg/L) of PAA up to 7 days. The stability and physical properties of collagen-free PAA-ACP solutions were also investigated. In our system, lowering PAA MW and increasing PAA concentration resulted in solutions with increasing stability. Over stable PAA-ACP solutions that fully inhibited mineralization of the collagen matrices were achieved using PAA 2k-50. Conversely, unstable solutions were obtained using high PAA MW at low concentrations. Nucleation and growth of significant amount of extrafibrillar minerals on the collagen fibrils was obtained using these solutions. In a wide range of combined MW and concentration of PAA we obtained intrafibrillar mineralization of collagen with hydroxyapatite crystals aligned parallel to the collagen fibril as in natural tissues. Intrafibrillar mineralization was correlated with PAA-ACP stability and growth of the PAA-ACP particles in solution. Our results support using PAA to surrogate NCPs function as selective inhibitors or promoters of biological mineralization and provide parameters to manufacture new biomimetic scaffolds and constructs for bone and dentin tissue engineering.


Antibacterial free fatty acids: activities, mechanisms of action and biotechnological potential.
Amongst the diverse and potent biological activities of free fatty acids (FFAs) is the ability to kill or inhibit the growth of bacteria. The antibacterial properties of FFAs are used by many organisms to defend against parasitic or pathogenic bacteria. Whilst their antibacterial mode of action is still poorly understood, the prime target of FFA action is the cell membrane, where FFAs disrupt the electron transport chain and oxidative phosphorylation. Besides interfering with cellular energy production, FFA action may also result from the inhibition of enzyme activity, impairment of nutrient uptake, generation of peroxidation and auto-oxidation degradation products or direct lysis of bacterial cells. Their broad spectrum of activity, non-specific mode of action and safety makes them attractive as antibacterial agents for various applications in medicine, agriculture and food preservation, especially where the use of conventional antibiotics is undesirable or prohibited. Moreover, the evolution of inducible FFA-resistant phenotypes is less problematic than with conventional antibiotics. The potential for commercial or biomedical exploitation of antibacterial FFAs, especially for those from natural sources, is discussed.


Silexan in generalized anxiety disorder: investigation of the therapeutic dosage range in a pooled data set.
Silexan, a special active substance produced from Lavandula angustifolia, is efficacious in subsyndromal anxiety at a dose of 80 mg/day, but its effective dosage in generalized anxiety disorder (GAD) has yet to be defined. In two double-blind, placebo-controlled trials, daily doses of 10, 40, 80, and 160 mg silexan were administered for 10 weeks. A total of 925 adults with GAD according to Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria and a Hamilton Anxiety Scale (HAMA) total score of at least 18 points were analyzed for efficacy. We assessed the change versus baseline for the HAMA and the Covi Anxiety Scale, the Clinical Global Impressions scale, the Sheehan Disability Scale, and the SF-36 health status questionnaire using analysis of variance and covariance. Silexan 160 mg/day was superior to placebo for all efficacy outcomes investigated, with responder rates exceeding 60% on the basis of HAMA and Clinical Global Impressions criteria. For the 80 mg/day dosage, superiority over placebo could be shown in one trial as well as in the pooled analysis. The risk of adverse events under silexan was similar to placebo for all dosages investigated. In GAD silexan 160 mg/day is efficacious whereas 80 mg/day may represent the lower end of the therapeutic range. Daily doses up to 160 mg were well tolerated.


Functional genomic analysis of constitutive and inducible defense responses to Fusarium verticillioides infection in maize genotypes with contrasting ear rot resistance.
Fusarium verticillioides causes ear rot in maize (Zea mays L.) and accumulation of mycotoxins, that affect human and animal health. Currently, chemical and agronomic measures to control Fusarium ear rot are not very effective and selection of more resistant genotypes is a desirable strategy to reduce contaminations. A deeper knowledge of molecular events and genetic basis underlying Fusarium ear rot is necessary to speed up progress in breeding for resistance. A next-generation RNA-sequencing approach was used for the first time to study transcriptional changes associated with F. verticillioides inoculation in resistant CO441 and susceptible CO354 maize genotypes at 72 hours post inoculation. More than 100 million sequence reads were generated for inoculated and uninoculated control plants and analyzed to measure gene expression levels. Comparison of expression levels between inoculated vs. uninoculated and resistant vs. susceptible transcriptomes revealed a total number of 6,951 differentially expressed genes. Differences in basal gene expression were observed in the uninoculated samples. CO441 genotype showed a higher level of expression of genes distributed over all functional classes, in particular those related to secondary metabolism category. After F. verticillioides inoculation, a similar response was observed in both genotypes, although the magnitude of induction was much greater in the resistant genotype. This response included higher activation of genes involved in pathogen perception, signaling and defense, including WRKY transcription factors and jasmonate/ethylene mediated defense responses. Interestingly, strong differences in expression between the two genotypes were observed in secondary metabolism category: pathways related to shikimate, lignin, flavonoid and terpenoid biosynthesis were strongly represented and induced in the CO441 genotype, indicating that selection to enhance these traits is an additional strategy for improving resistance against F. verticillioides infection. The work demonstrates that the global transcriptional analysis provided an exhaustive view of genes involved in pathogen recognition and signaling, and controlling activities of different TFs, phytohormones and secondary metabolites, that contribute to host resistance against F. verticillioides. This work provides an important source of markers for development of disease resistance maize genotypes and may have relevance to study other pathosystems involving mycotoxin-producing fungi.


Stacking-Based Deep Neural Network: Deep Analytic Network for Pattern Classification.
Stacking-based deep neural network (S-DNN) is aggregated with pluralities of basic learning modules, one after another, to synthesize a deep neural network (DNN) alternative for pattern classification. Contrary to the DNNs trained from end to end by backpropagation (BP), each S-DNN layer, that is, a self-learnable module, is to be trained decisively and independently without BP intervention. In this paper, a ridge regression-based S-DNN, dubbed deep analytic network (DAN), along with its kernelization (K-DAN), are devised for multilayer feature relearning from the pre-extracted baseline features and the structured features. Our theoretical formulation demonstrates that DAN/K-DAN relearn by perturbing the intra/interclass variations, apart from diminishing the prediction errors. We scrutinize the DAN/K-DAN performance for pattern classification on datasets of varying domains-faces, handwritten digits, generic objects, to name a few. Unlike the typical BP-optimized DNNs to be trained from gigantic datasets by GPU, we reveal that DAN/K-DAN are trainable using only CPU even for small-scale training sets. Our experimental results show that DAN/K-DAN outperform the present S-DNNs and also the BP-trained DNNs, including multiplayer perceptron, deep belief network, etc., without data augmentation applied.


Effects of lane departure warning on police-reported crash rates.
To evaluate the effects of lane departure warning (LDW) on single-vehicle, sideswipe, and head-on crashes. Police-reported data for the relevant crash types were obtained from 25 U.S. states for the years 2009-2015. Observed counts of crashes with fatalities, injuries, and of all severities for vehicles with LDW were compared with expected counts based on crash involvement rates for the same passenger vehicles without LDW, with exposure by vehicle series, model year, and lighting system standardized between groups. For relevant crashes of all severities and those with injuries, Poisson regression was used to estimate the benefits of LDW while also controlling for demographic variables; fatal crashes were too infrequent to be modeled. Without accounting for driver demographics, vehicles with LDW had significantly lower involvement rates in crashes of all severities (18%), in those with injuries (24%), and in those with fatalities (86%). Adding controls for driver demographics in the Poisson regression reduced the estimated benefit of LDW only modestly in crashes of all severities (11%, p < 0.05) and in crashes with injuries (21%, p < 0.07). Lane departure warning is preventing the crash types it is designed to address, even after controlling for driver demographics. Results suggest that thousands of lives each year could be saved if every passenger vehicle in the United States were equipped with a lane departure warning system that performed like the study systems. Purchase of LDW should be encouraged, and, because drivers do not always keep the systems turned on, future efforts should focus on designing systems to encourage greater use and educating consumers about the benefits of using the systems.


Effect of iron depletion on cardiovascular risk factors: studies in carbohydrate-intolerant patients.
Controversy surrounds the role of iron (Fe) in atherosclerosis (ASCVD), mainly due to the inaccuracy of assessing body Fe stores with serum ferritin and transferrin saturation. Quantitative phlebotomy was used to test whether or not (a) Fe stores are increased in individuals at high risk for ASCVD and (b) Fe depletion to near-deficiency (NID) levels is associated with reduction of risk factors for ASCVD. Thirty-one carbohydrate-intolerant subjects completed the study. Fe stores were within normal limits (1.5 +/- 0.1 g). At NID, a significant increase of HDL-cholesterol (p < 0.001) and reductions of blood pressure (p < 0.001), total and LDL-cholesterol (p < 0.001), triglyceride (p < 0.001), fibrinogen (p < 0.001) and glucose and insulin responses to oral glucose loading (p < 0.001) were noted, while homocysteine plasma concentration remained unchanged. These effects were largely reversed by a 6-month period of Fe repletion with reinstitution of Fe sufficiency. Thus, although individuals at high risk for ASCVD are not Fe-overloaded, they seem to benefit, metabolically and hemodynamically, from lowering of body Fe to levels commonly seen in premenopausal females.


New anthocyanins from purple pods of pea (Pisum spp.).
Two new anthocyanins were isolated from purple pods of pea (Pisum spp.). Their structures were identified as delphinidin 3-xylosylgalactoside-5-acetylglucoside and its deacetylated derivative by the usual chemical degradation methods and by spectroscopic methods such as UV-VIS, MS and NMR. Both pigments showed moderate stability and antioxidative activity in a neutral aqueous solution.


Acupuncture upregulates G protein coupled activity in SAMP8 mice.
Transmembrane and intracellular signal transduction of G protein is closely related to the pathophysiology of Alzheimer's disease (AD). To explore the effects of Sanjiao acupuncture on G protein signal transduction pathways in the pathogenesis of AD. 36 senescence-accelerated (SAM) prone 8 mice were divided into three groups that remained untreated (SAMP8, n=12) or received Sanjiao acupuncture (SAMP8+SA, n=12) or control acupuncture (SAMP8+CA, n=12). An additional control group of SAM resistant 1 mice was included (SAMR1 group, n=12). Morris water maze tests were used to investigate learning and memory abilities. Immunoprecipitation and Western blotting were used to study expression of G protein subunits and their activities in the cortex/hippocampus. Behavioural analysis showed that acupuncture attenuated the severe cognitive deficits observed in untreated/CA-treated SAMP8 mice. The findings of the G protein activation assays via immunoprecipitation and Western blots were that the physiologically coupled activation rate (PCAR) and maximal coupled activation rate (MCAR) of Gαs and Gαi were decreased in the cortex of SAMP8 vs SAMR1 mice. Sanjiao acupuncture induced an upregulation in the PCAR of Gαs and Gαi. In the hippocampus of untreated SAMP8 mice, the PCAR of Gαs and MCAR of both Gαs and Gαi declined, and Sanjiao acupuncture was associated with an upregulation in the MCAR of Gαs and Gαi. There were no significant differences in Gαs and Gαi expression between the groups. Sanjiao acupuncture attenuates cognitive deficits in a mouse model of AD via upregulation of G protein activity and stabilisation of the cellular signal.


Low-dose glucocorticoids in hyperandrogenism.
To investigate the effect of low-doses of glucocorticoids on androgen and cortisol secretion during the course of the day, we evaluated clinical signs of hyperandrogenism and total, free and bioavailable testosterone, SHBG, and cortisol following two different protocols: A) fourteen patients received betAmethasone 0.6 mg/day (n=8) or methylprednisolone 4 mg/day (n=6), as single daily oral dose at 11.00 PM, during 30 days, B) fourteen patients were evaluated under betamethasone 0.3 mg in a single daily dose at 11.00 PM during six months, 11 out of whom were re-evaluated six months later. Twenty eight women with hyperandrogenism were included and seven normal females were used as control. Blood samples were taken in follicular phase at 8 AM and 7 PM to determine SHBG, cortisol, total, free and bioavailable testosterone. In both protocols, a significant morning and evening decrease in cortisol and testosterone (p<0.05 to < 0.01), which was more marked with betamethasone (p<0.05), was shown. In protocol B, morning SHBG levels showed a significant increase (p<0.05) and betamethasone also improved clinical hyperandrogenism along the trial. Although morning and evening cortisol significantly decreased during treatment, no side effects were reported. The 11 patients reevaluated after therapy withdrawal, showed a rise in serum total testosterone and its fractions to pre-treatment values and a normalization of cortisol levels. It is concluded that glucocorticoids in low-doses effectively normalize serum androgens, independently of their origin. They may be used therapeutically, mainly whenever a hyperandrogenic woman presents with cycle irregularities or seeking fertility.


The pediatrician's role in the evaluation and preparation of pediatric patients undergoing anesthesia.
Pediatricians play a key role in helping prepare patients and families for anesthesia and surgery. The questions to be answered by the pediatrician fall into 2 categories. The first involves preparation: is the patient in optimal medical condition for surgery, and are the patient and family emotionally and cognitively ready for surgery? The second category concerns logistics: what communication and organizational needs are necessary to enable safe passage through the perioperative process? This revised statement updates the recommendations for the pediatrician's role in the preoperative preparation of patients.


Underestimation of hepatic glucose production by radioactive and stable tracers.
Although negative hepatic glucose production rates are physiologically impossible, they have been observed when hepatic glucose production is measured with the tracer-dilution technique during the hyperinsulinemic, euglycemic glucose clamp. Because hepatic glucose production is determined from the difference between tracer-derived glucose disposal and the known exogenous glucose infusion rate, the negative values for hepatic glucose production must result from an underestimation of glucose disposal by the tracer technique. In the current investigation, tracer-derived glucose disposal was measured in 25 subjects undergoing hyperinsulinemic, euglycemic clamps. Glucose disposal was measured with both radioactive and stable isotopes that utilize different methodologies, to determine whether discriminant metabolism of the isotopes versus methodological error leads to underestimation of tracer-derived glucose disposal. Both the radioactive and stable methodologies underestimated the exogenous glucose infusion rate during the hyperinsulinemic euglycemic clamp by 27 and 17%, respectively. Mean hepatic glucose production was -2.1 +/- 0.2 and -1.3 +/- 0.2 mg X kg-1 X min-1 as determined by the radioactive and stable isotope methodologies, respectively. Methodological error was an unlikely cause of this underestimation because it occurred with two different methodologies. The most likely explanation for underestimated rates of glucose disposal determined by the two types of isotope methodologies is discrepant metabolism of glucose tracers in comparison with unlabeled glucose.


Informational support to family members of intensive care unit patients: the perspectives of families and nurses.
The receiving information about the patients hospitalized in the intensive care unit is classified among the most important needs of the family members of such patients. Meeting the informational needs of families is a major goal for intensive care workers. Delivering honest, intelligible and effective information raises specific challenges in the stressful setting of the intensive care unit (ICU). The aim of this qualitative study was to explain perspectives of families of Intensive Care Unit patients and nurses about informational support. Using a conventional content analysis approach, semi-structured interviews were conducted with participants to explore their perspectives of providing informational support to families of ICU patients. A purposeful sampling method was used to recruit nineteen family members of thirteen patients hospitalized in the ICU and twelve nurses from three teaching hospitals. In general, 31 persons participated in this study. Data collection continued to achieve data saturation. A conventional content analysis of the data produced three categories and seven sub-categories. The three main categories were as followed, a) providing information, b) handling information and c) using information. Providing information had three sub-categories consisting of "receiving admission news", "receiving truthful and complete information" and receiving general information. Handling information had two sub-categories consisting "keeping information" and "gradual revelation". Lastly, using information has two sub-categories consisting of "support of patient" and "support of family members". The results of this study revealed perspectives of families of Intensive Care Unit patients and nurses about informational support. It also determines the nurses' need to know more about the influence of their supportive role on family's ICU patients informing. In addition, the results of present study can be used as a basis for further studies and for offering guidelines about informational support to the families of the patients hospitalized in the ICU.


[A suction irrigation endoscope for surgery of the sinuses and skull base (author's transl)].
For optimal transnasal surgery of the paranasal sinuses or of the skull base, free endoscopic exposure of the operative field is necessary. In our experience, use of instrumentation adjacent to the endoscope allows better removal of tissue specimens in contrast to the use of smaller forceps placed through the endoscope itself. A suction irrigation endoscope is described which allows manipulation under direct endoscopic control, but without disturbance by blood or secretions.


Constitutive production of a non-interferon macrophage activating factor by HTLV-transformed T cells.
Culture supernatants of several human T cell leukemia virus infected cell lines were assessed for the induction of tumoricidal activity against human melanoma target cells in a 72-hr assay. Two cell lines, MT-2 and C10/MJ2, were found to produce high levels of MAF activity constitutively. However, the MT-2 cell line, unlike C10/MJ2, produced little interferon. MT-2 MAF was distinct from interferon-gamma by a number of biochemical and serological criteria. The MAF activity was associated with a non-glycosylated, acid stable protein which has an apparent molecular weight of 55,000 and an isoelectric point of 5.5. Furthermore, this molecule is not related to any of the known lymphokines or cytokines such as the interleukins or colony stimulating factors.


Effects of sodium bentonite, buffer salts, cement kiln dust and clinoptilolite on rumen characteristics of beef steers fed a high roughage diet.
Five ruminally cannulated Hereford steers (average weight 340 kg) were used in a 5 x 5 Latin square design experiment to examine the influences of dietary sodium bentonite (SB), McDougall's buffer salts (MB), cement kiln dust (KD) and clinoptilolite (CLN) on rumen liquid dilution rate, fermentation patterns and roughage intake. Treatments were administered as 350-g additions to a basal protein/energy supplement of 2.62 kilograms. The basal diet of cottonseed hulls was offered ad libitum. Mineral analyses of the four treatment substances revealed fairly high levels of potentially toxic trace minerals (e.g., Pb, Cd and F) in the KD. Intake of cottonseed hulls was not significantly affected by treatment but tended to be higher for MB-supplemented steers (7.19 kg) than for those given the control or other treatments (average 6.72 kg). Some rejection of supplements containing MB and KD was noted. At e hr after supplement feeding rumen pH of MB-treated steers (6.68) was higher (P less than .05) than that of steers fed KD (6.50) and CLN (6.44) but was not different from that od control or SB-treated steers (6.56). Seven hours after supplement was offered rumen pH of MB steers (6.74 was higher (P less than .05) for steers fed MB, KD and CLN than for SB-an control-fed steers. Volatile fatty acids, rumen ammonia, osmolality and protein and ash contents of a crude bacterial fraction were not significantly influenced by mineral compounds, nor was rumen liquid volume. Values for rumen liquid volume were as follows: MB (60.27 liters), SB (50.43 liters), CLN (45.53 liters), control (43.14 liters) and KD (41.81 liters). Liquid dilution rate (percentage per hour) was high for steers on all treatments (average of 9.11) and was not significantly altered by mineral supplementation.


Characterization of the cholera toxin receptor on Balb/c 3T3 cells as a ganglioside similar to, or identical with, ganglioside GM1. No evidence for galactoproteins with receptor activity.
Balb/c 3T3 cells contain a large number [(0.8-1.6) x 10(6)] of high-affinity (half-maximal binding at 0.2 nM) binding sites for cholera toxin that are resistant to proteolysis, but are quantitatively extracted with chloroform/methanol. The following evidence rigorously establishes that the receptor is a ganglioside similar to, or identical with, ganglioside GM1 by the galactose oxidase/NaB3H4 technique on intact cells was inhibited by cholera toxin. (2) Ganglioside GM1 was specifically adsorbed from Nonidet P40 extracts of both surface- (galactose oxidase/NaB3H4 technique) and metabolically ([1-14C]palmitate) labelled cells in the presence of cholera toxin, anti-toxin and Staphylococcus aureus. (3) Ganglioside GM1 was the only ganglioside labelled when total cellular gangliosides separated on silica-gel sheets were overlayed with 125I-labelled cholera toxin, although GM3 and GD1a were the major gangliosides present. In contrast no evidence for a galactoprotein with receptor activity was obtained. Cholera toxin did not protect the terminal galactose residues of cell-surface glycoproteins from labelling by the galactose oxidase/NaB3H4 technique. No toxin-binding proteins could be identified in Nonidet P40 extracts of [35S]-methionine-labelled cells by immunochemical means. After sodium dodecyl sulphate/polyacrylamide-gel electrophoresis none of the major cellular galactoproteins identified by overlaying gels with 125I-labelled ricin were able to bind 125I-labelled cholera toxin. It is concluded that the cholera toxin receptor on Balb/c 3T3 cells is exclusively ganglioside GM1 (or a related species), and that cholera toxin can therefore be used to probe the function and organisation of gangliosides in these cells as previously outlined [Critchley, Ansell, Perkins, Dilks & Ingram (1979) J. Supramol. Struct. 12, 273-291].


Tricyclic antidepressant plasma level monitoring: an improvement over the dose-response approach.
Monitoring plasma levels of tricyclic antidepressants (TCAs) can facilitate treatment response by providing objective guidelines for dosage adjustment. It provides a means of assessing the patient's compliance and ensuring the maintenance of an adequate medication trial while avoiding the complications of drug toxicity. The traditional dose-response method of medication adjustment depends on several assumptions that are not met in the case of TCAs, which makes plasma monitoring particularly useful for patients receiving most TCAs.


Diagnosis of Spotted Fever Group Rickettsioses in U.S. Travelers Returning from Africa, 2007-2016.
Spotted fever group rickettsioses (SFGRs), such as African tick bite fever (ATBF), are among the most commonly diagnosed diseases for ill travelers returning from southern Africa. We summarized demographic, clinical, and diagnostic features of imported SFGR cases in U.S. travelers returning from Africa who had laboratory specimens submitted to the Centers for Disease Control and Prevention. Diagnosis of SFGR was performed by indirect immunofluorescence antibody assay, immunohistochemical staining, polymerase chain reaction (PCR), or culture. Cases were defined as probable SFGR, confirmed SFGR, or confirmed ATBF. Clinical and epidemiological categorical variables were described as counts and proportions; continuous variables were described using geometric mean titers, median, and range. One hundred and twenty-seven patients satisfied laboratory criteria for confirmed or probable SFGR. Fever was the most common symptom (N = 88; 69%), followed by ≥ 1 eschars (N = 70; 55%). Paired serums were submitted for 36 patients (28%); 12 patients (33%) had nonreactive initial serum sample but converted to a titer ≥ 64 with the convalescent sample. Twenty-seven patients (21%) had infection with Rickettsia africae based on PCR analysis of eschar swab (N = 8) or biopsy (N = 23). Fifteen patients had eschar biopsy or swab samples and serum sample(s) submitted together; 9 (60%) had PCR-positive eschar results and nonreactive acute serology. Health-care providers should consider SFGR when evaluating patients for a febrile illness with eschar and compatible foreign travel history. Polymerase chain reaction testing of eschar biopsies or swabs provides a confirmed diagnosis in early stages of disease; eschar swabs or biopsies are an underutilized diagnostic technique.


Artemether Attenuates Gut Barrier Dysfunction and Intestinal Flora Imbalance in High-Fat and High-Fructose Diet-Fed Mice.
Intestinal inflammation is a key determinant of intestinal and systemic health, and when our intestines are damaged, there is disruption of the intestinal barrier, which in turn induces a systemic inflammatory response. However, the etiology and pathogenesis of inflammatory diseases of the intestine are still not fully understood. Artemether (ART), one of the artemisinin derivatives, has been widely used to treat malaria. Nevertheless, the effect of ART on intestinal inflammation remains unclear. The present study intended to elucidate the potential mechanism of ART in diet-induced intestinal injury. A high-fat and high-fructose (HFHF) diet-induced mouse model of intestinal injury was constructed, and the mice were treated with ART to examine their role in intestinal injury. RT-qPCR, Western blotting, immunohistochemical staining, and 16S rRNA gene sequencing were used to investigate the anti-intestinal inflammation effect and mechanism of ART. The results indicated that ART intervention may significantly ameliorate the intestinal flora imbalance caused by the HFHF diet and alleviate intestinal barrier function disorders and inflammatory responses by raising the expression of tight junction proteins ZO-1 and occludin and decreasing the expression of pro-inflammatory factors TNF-α and IL-1β. Moreover, ART intervention restrained HFHF-induced activation of the TLR4/NF-κB p65 pathway in colon tissue, which may be concerned with the potential protective effect of ART on intestinal inflammation. ART might provide new insights into further explaining the mechanism of action of other metabolic diseases caused by intestinal disorders.


A phase II study of sorafenib in patients with platinum-pretreated, advanced (Stage IIIb or IV) non-small cell lung cancer with a KRAS mutation.
Sorafenib inhibits the Ras/Raf pathway, which is overactive in cancer patients with a KRAS mutation. We hypothesized that patients with non-small cell lung cancer (NSCLC) with KRAS mutation will benefit from treatment with sorafenib. In this phase II study, patients with KRAS-mutated, stage IIIb or IV NSCLC that progressed after at least one platinum-containing regimen were treated with sorafenib. Treatment consisted of sorafenib 400 mg twice daily until disease progression or unacceptable toxicity. Pretreatment serum from each patient was obtained to predict outcome using a proteomic assay (VeriStrat). Primary endpoint was disease control rate (DCR) at 6 weeks. Fifty-nine patients were entered between May 2010 and February 2011. Fifty-seven patients started sorafenib. Mean age was 58.5 (SD = ±8.1) years, 16 male/41 female, Eastern Cooperative Oncology Group (ECOG) performance status (PS) 0/1/2 24/30/3. At 6 weeks, 5 partial response, 25 stable disease, and 27 progressive disease were observed; DCR was 52.6%. Median duration of treatment was 9 weeks. The median progression-free survival (PFS) was 2.3 months and median overall survival (OS) was 5.3 months. Patients with a prediction of good prognosis according to VeriStrat serum proteomics assay showed a significantly superior PFS [HR, 1.4; 95% confidence interval (CI), 1.0-1.9] but not OS (HR, 1.3; 95% CI, 0.9-1.7). Sorafenib-related grade III/IV toxicity was reported in 10 patients (17.5%); all but one patient experienced grade III skin toxicity (14.0%) or grade III gastrointestinal toxicity (8.8%). Treatment with sorafenib has relevant clinical activity in patients with NSCLC harboring KRAS mutations. Further randomized study with this agent is warranted as single-agent or combination therapy.


Expression of activated PKC epsilon (PKC epsilon) protects the ischemic heart, without attenuating ischemic H(+) production.
PKC epsilon is a PKC isoform that translocates during preconditioning and may mediate cardioprotection. To investigate whether PKC epsilon activation is cardioprotective, Langendorff-perfused hearts from wild-type (WT) mice and from mice expressing constitutively active mutant PKC epsilon were subjected to 20 min ischemia and 40 min reperfusion while(31)P NMR spectra were acquired. Pre-ischemic glycogen levels were similar in WT and PKC epsilon hearts. During ischemia, ATP fell less in PKC epsilon than in WT hearts. Ischemic intracellular pH, however, was similar in WT and PKC epsilon hearts. During reperfusion, recovery of contractile function and ATP were greater in PKC epsilon than WT hearts. In conclusion, expression of activated PKC epsilon protected hearts from post-ischemic energetic and contractile dysfunction, consistent with the proposed cardioprotective role of PKC epsilon. Protection occurred in the PKC epsilon hearts without attenuation of ischemic H(+) production, implying that, at least in this ischemic model, reduced acidification during ischemia is not necessary for cardioprotection.


[Indices of fast acoustic adaptation in unilateral recording of short latent acoustic potentials in people with normal hearing and chronic functional voice disturbances].
The authors presented the results of the study of the central parts of the acoustic analyzer using the method of recording the short latent acoustic evoked potentials (SAEP), with stimulus frequency of 21.1 and 90 Hz, in the persons suffered from chronic functional voice disturbances (CFVD). A total of 67 persons aged from 23 to 45 years with normal hearing function were examined; among them, the normal voice forming and the acoustic apparatus were found in 38, and CFVD was detected in 29 persons. The state of the central parts of the acoustic analyzer examined by using a method of fast acoustic adaptation (FAA) during recording the SAEP points to the interrelation between the acoustic and vocal systems on the level of brainstem. It should be mentioned that investigating the brainstem responses to auditory stimuli, especially of different frequency, is helpful to reveal the preclinical disturbances in the functional state of the central nervous system, and brainstem structures of the acoustic analyzer, in particular. So, it contributes to early diagnosis of these affections in the persons with voice and hearing pathology. The study of the central mechanisms of the interrelations between the acoustic and voice forming systems contributes to new progressive achievements in phonoaudiology and elaboration of new approaches in diagnosis and treating the voice and acoustic disturbances, which remain one of the burning problems in present otorhinolaryngology.


Macrophage Transplantation Fails to Improve Repair of Critical-Sized Calvarial Defects.
Over 500,000 bone grafting procedures are performed every year in the United States for neoplastic and traumatic lesions of the craniofacial skeleton, costing $585 million in medical care. Current bone grafting procedures are limited, and full-thickness critical-sized defects (CSDs) of the adult human skull thus pose a substantial reconstructive challenge for the craniofacial surgeon. Cell-based strategies have been shown to safely and efficaciously accelerate the rate of bone formation in CSDs in animals. The authors recently demonstrated that supraphysiological transplantation of macrophages seeded in pullalan-collagen composite hydrogels significantly accelerated wound healing in wild type and diabetic mice, an effect mediated in part by enhancing angiogenesis. In this study, the authors investigated the bone healing effects of macrophage transplantation into CSDs of mice. CD1 athymic nude mice (60 days of age) were anesthetized, and unilateral full-thickness critical-sized (4 mm in diameter) cranial defects were created in the right parietal bone, avoiding cranial sutures. Macrophages were isolated from FVB-L2G mice and seeded onto hydroxyapatite-poly (lactic-co-glycolic acid) (HA-PLGA) scaffolds (1.0 × 10 cells per CSD). Scaffolds were incubated for 24 hours before they were placed into the CSDs. Macrophage survival was assessed using three-dimensional in vivo imaging system (3D IVIS)/micro-CT. Micro-CT at 0, 2, 4, 6, and 8 weeks was performed to evaluate gross bone formation, which was quantified using Adobe Photoshop. Microscopic evidence of bone regeneration was assessed at 8 weeks by histology. Bone formation and macrophage survival were compared at each time point using independent samples t tests. Transplantation of macrophages at supraphysiological concentration had no effect on the formation of bones in CSDs as assessed by either micro-CT data at any time point analyzed (all P > 0.05). These results were corroborated by histology. 3D IVIS/micro-CT demonstrated survival of macrophages through 8 weeks. Supraphysiologic delivery of macrophages to CSDs of mice had no effect on bone formation despite survival of transplanted macrophages through to 8 weeks posttransplantation. Further research into the physiological effects of macrophages on bone regeneration is needed to assess whether recapitulation of these conditions in macrophage-based therapy can promote the healing of large cranial defects.


Ror1 is a pseudokinase that is crucial for Met-driven tumorigenesis.
The human kinome includes Ror1, a poorly characterized orphan receptor. Here we report the findings of an investigation of Ror1 contributions to cancer, undertaken through an integrated screening of 43 cancer cell lines where we measured protein expression, tyrosine phosphorylation, and growth response following RNAi-mediated Ror1 suppression. Ror1 was expressed in approximately 75% of the cancer cell lines without apparent histotype distribution. Gastric carcinoma cells (HS746T) and non-small cell lung carcinoma cells (NCI-H1993) exhibited high levels of Ror1 tyrosine phosphorylation, and Ror1 suppression caused growth inhibition. Biochemical assays revealed unexpectedly that Ror1 is a pseudokinase that is devoid of catalytic activity. Intriguingly, the two cell lines featuring tyrosine-phosphorylated Ror1 both exhibited amplification and activation of the Met oncogene. Ror1 phosphorylation was abrogated by Met inhibition, indicating Met-dependent transphosphorylation of Ror1. Conversely, Ror1 was not transphosphorylated by other constitutively active tyrosine kinases, including EGFR and ErbB2. Constitutive silencing of Ror1 in HS746T and NCI-H1993 carcinoma cells impaired proliferation in vitro and induced a dramatic inhibition of tumorigenesis in vivo. Together, our findings suggest a critical role for Ror1 in malignant phenotypes sustained by the Met oncogene.


Endogenous nitrogen excretion by red kangaroos (Macropus rufus): effects of animal age and forage quality.
Red kangaroos (Macropus rufus) are large (>20 kg) herbivorous marsupials common to arid and semiarid Australia. The population dynamics of red kangaroos are linked with environmental factors, operating largely through juvenile survival. A crucial period is the young-at-foot (YAF) stage, when juveniles have permanently left the mother's pouch but still take milk from a teat in the pouch. Forage quantity and quality have been implicated in drought-related mortalities of juvenile kangaroos. Here we compared how forage quality affected nitrogen (N) intake and excretion by YAF, weaned, and mature, nonlactating female red kangaroos. On high-quality forage (chopped lucerne hay, Medicago sativa) low in neutral-detergent fiber (43%+/-1%) and high in N (2.9%+/-0.1%), YAF and weaned kangaroos had ideal growth rates and retained 460-570 mg dietary N kg(-0.75) d(-1). But on poor-quality forage (chopped oaten hay, Avena sativa) high in neutral-detergent fiber (64%+/-1%) and low in N (0.9%+/-0.1%), YAF and weaned kangaroos could not sustain growth and were in negative N balance at -103+/-26 mg and -57+/-31 mg N kg(-0.75) d(-1), respectively. Notably, the YAF kangaroos excreted 64% of their truly digestible N intake from forage as nondietary fecal N (NDFN). By weaning age, the situation had improved, but the juveniles still lost 40% of their truly digestible N intake as NDFN compared with only 30% by the mature females. Our findings support field observations that forage quality, and not just quantity, is a major factor affecting the mortality of juvenile red kangaroos during drought.


Germinal mosaicism for a deletion of the FMR1 gene leading to fragile X syndrome.
Aberrant CGG trinucleotide amplification within the FMR1 gene, which spans approximately 38 Kb of genomic DNA is almost always what leads to fragile X syndrome (FXS). However, deletions of part or the entire FMR1 gene can also cause FXS. Both CGG amplification-induced silencing and deletions result in the absence of the FMR1 gene product, FMRP. Here, we report a rare case of germinal mosaicism of a deletion encompassing approximately 300 Kb of DNA, which by removing the entire FMR1 gene led to FXS. The male proband, carrying the deletion, presented in clinic with the typical features of FXS. His mother was analyzed by FISH on metaphase chromosomes with cosmid probe c22.3 spanning the FMR1 locus, and she was found not to carry the deletion on 30 analyzed cells from peripheral blood lymphocytes. Prenatal examination of the mother's third pregnancy showed that the male fetus also had the same deletion as the proband. Following this prenatal diagnosis, FISH analysis in the mother was expanded to 400 metaphases from peripheral lymphocytes, and a heterozygous FMR1 deletion was found in three. Although this result could be considered questionable from a diagnostic point of view, it indicates that the deletion is in the ovary's germinal cells.


The effect of boundary conditions on epicardial potential distributions.
This study presents a comparison of semi-analytical and numerical solution techniques for solving the passive bidomain equation in simple tissue geometries containing a region of subendocardial ischaemia. When the semi-analytical solution is based on Fourier transforms, recovering the solution from the frequency domain via fast Fourier transforms imposes a periodic boundary condition on the solution of the partial differential equation. On the other hand, the numerical solution uses an insulation boundary condition. When these techniques are applied to calculate the epicardial surface potentials, both yield a three well potential distribution which is identical if fibre rotation within the tissue is ignored. However, when fibre rotation is included, the resulting three-well distribution rotates, but through different angles, depending on the solution method. A quantitative comparison between the semi-analytical and numerical solutiontechniques is presented in terms of the effect fibre rotation has on the rotation of the epicardial potential distribution. It turns out that the Fourier transform approach predicts a larger rotation of the epicardial potential distribution than the numerical solution. The conclusion from this study is that it is not always possible to use analytical or semi-analytical solutions to check the accuracy of numerical solution procedures. For the problem considered here, this checking is only possible when it is assumed that there is no fibre rotation through the tissue.


Endotheliopathy of Trauma is an on-Scene Phenomenon, and is Associated with Multiple Organ Dysfunction Syndrome: A Prospective Observational Study.
Trauma patients are vulnerable to coagulopathy and inflammatory dysfunction associated with endotheliopathy of trauma (EoT). In vitro evidence has suggested that tranexamic acid (TXA) may ameliorate endotheliopathy. We aimed to investigate how soon after injury EoT occurs, its association with multiple organ dysfunction syndrome (MODS), and whether TXA ameliorates it. A prospective observational study included 91 trauma patients enrolled within 60 min of injury and 19 healthy controls. Blood was sampled on enrolment and again 4 to 12 h later. ELISAs measured serum concentrations of syndecan-1 and thrombomodulin as biomarkers of EoT. MODS was compared between groups according to biomarker dynamics: persistently abnormal; abnormal to normal; and persistently normal. Timing of EoT was estimated by plotting biomarker data against time, and then fitting generalized additive models. Biomarker dynamics were compared between those who did or did not receive prehospital TXA. Median age was 38 (interquartile range [IQR] 24-55) years; 78 of 91 were male. Median injury severity score (ISS) was 22 (IQR 12-36). EoT was estimated to occur at 5 to 8 min after injury. There were no significant differences in ISS between those with or without prehospital EoT. Forty-two patients developed MODS; 31 of 42 with persistently abnormal; 8 of 42 with abnormal to normal; and 3 of 42 with persistently normal biomarkers; P < 0.05. There were no significant differences between TXA and non-TXA groups. EoT was present at the scene of injury. MODS was more likely when biomarkers of EoT were persistently raised. There were no significant differences between TXA and non-TXA groups. Prehospital interventions aimed at endothelial restoration may represent a clinically meaningful target for prehospital resuscitation.


Interactions of the platelets in paroxysmal nocturnal hemoglobinuria with complement. Relationship to defects in the regulation of complement and to platelet survival in vivo.
The blood cells of patients with paroxysmal nocturnal hemoglobinuria (PNH) have abnormal interactions with complement. The activity of the alternative pathway C3 convertase on the platelets of 9 out of 19 patients with PNH was elevated. 10 patients had C3 convertase activity within the normal range even though 80-95% of their platelets lacked the complement regulatory protein decay accelerating factor (DAF) that is absent from the affected blood cells in PNH. PNH and normal platelets released factor H when C3 was bound to their surfaces. This may account for the apparent regulation of C3 convertase activity on platelets that lack DAF. The abnormal uptake of the membrane attack complex of complement by PNH III erythrocytes was not seen in PNH platelets. 111Indium-labeled platelet survival times were normal in five of eight patients, which suggests that the lack of the membrane attack complex defect results in normal platelet survival in PNH.


The trophectoderm acts as a niche for the inner cell mass through C/EBPα-regulated IL-6 signaling.
IL-6 has been shown to be required for somatic cell reprogramming into induced pluripotent stem cells (iPSCs). However, how Il6 expression is regulated and whether it plays a role during embryo development remains unknown. Here, we describe that IL-6 is necessary for C/EBPα-enhanced reprogramming of B cells into iPSCs but not for B cell to macrophage transdifferentiation. C/EBPα overexpression activates both Il6 and Il6ra genes in B cells and in PSCs. In embryo development, Cebpa is enriched in the trophectoderm of blastocysts together with Il6, while Il6ra is mostly expressed in the inner cell mass (ICM). In addition, Il6 expression in blastocysts requires Cebpa. Blastocysts secrete IL-6 and neutralization of the cytokine delays the morula to blastocyst transition. The observed requirement of C/EBPα-regulated IL-6 signaling for pluripotency during somatic cell reprogramming thus recapitulates a physiologic mechanism in which the trophectoderm acts as niche for the ICM through the secretion of IL-6.


Thiol-reactive dyes for fluorescence labeling of proteomic samples.
Covalent derivatization of proteins with fluorescent dyes prior to separation is increasingly used in proteomic research. This paper examines the properties of several commercially available iodoacetamide and maleimide dyes and discusses the conditions and caveats for their use in labeling of proteomic samples. The iodoacetamide dyes BODIPY TMR cadaverine IA and BODIPY Fl C(1)-IA were highly specific for cysteine residues and showed little or no nonspecific labeling even at very high dye:thiol ratios. These dyes also showed minimal effects on pI's of standard proteins. Some iodoacetamide dyes, (5-TMRIA and eosin-5-iodoacetamide) and some maleimide dyes (ThioGlo I and Rhodamine Red C(2) maleimide) exhibited nonspecific labeling at high dye:thiol ratios. Labeling by both iodoacetamide and maleimide dyes was inhibited by tris(2-carboxyethyl)phosphine (TCEP); interactions between TCEP and dye were also observed. Thiourea, an important component of sample solubilization cocktails, inhibited labeling of proteins with iodoacetamide dyes but not with maleimide dyes. Maleimide dyes may serve as an alternative for labeling proteins where it is essential to have thiourea in the solubilization buffer. Covalent derivatization by BODIPY TMR cadaverine IA, BODIPY Fl C(1)-IA or Rhodamine Red C(2) maleimide was also demonstrated to be compatible with in-gel digestion and peptide mass fingerprinting by matrix assisted laser desorption/ionization-mass spectrometry and allowed successful protein identification.


The effect of mesenchymal stem cells on osteoclast precursor cell differentiation.
Transplantation of mesenchymal stem cells (MSCs) has been extensively studied in the field of regenerative medicine. Bone regeneration is achieved via the interaction of osteoblasts and osteoclasts. However, the influence of MSCs on osteoclasts is unknown. The purpose of this study was to investigate the effect of MSCs on the expression of genes for osteoclast differentiation factors using qPCR after indirect co-culture of MSCs and RAW264 cells. The numbers of osteoclasts after addition of soluble receptor activator of nuclear factor kappa B (NF-κB) ligand (sRANKL) were also compared. Expression of osteoprotegerin (OPG) by MSCs was significantly elevated in co-culture over time. The differentiation of RAW264 cells into mature osteoclasts following addition of sRANKL was significantly inhibited by co-culture with MSCs. Expression of RANK, colony stimulating factor 1 receptor, NF-κB, and nuclear factor of activated T-cell cytoplasmic 1 in RAW264 cells was significantly inhibited by co-culture with MSCs. Expression of OPG protein was higher in co-culture with RAW264 cells than in MSCs alone, and the expression level was clearly higher than that of RANKL. MSCs appeared to inhibit osteoclast differentiation via expression of OPG.


Nuclear factor-kappa B contributes to excitotoxin-induced apoptosis in rat striatum.
Excitotoxin-induced destruction of striatal neurons, proposed as a model of Huntington's disease, involves a process having the biochemical stigmata of apoptosis. Recent studies suggested that transcription factor nuclear factor (NF)-kappa B may be involved in excitotoxicity. To further analyze the contribution of NF kappa B to excitotoxic neuronal death in vivo, changes in binding activities of NF kappa B and other transcription factors as well as the consequences of inhibiting NF kappa B nuclear translocation were measured after the infusion of quinolinic acid (120 nmol) into rat striatum. Internucleosomal DNA fragmentation and terminal transferase-mediated dUTP digoxigenin nick end labeling-positive nuclei appeared 12 hr later and intensified over the next 12 hr. NF kappa B binding activity increased several-fold from 2 to 12 hr, then gradually declined during the next 12 hr. Other transcription factor changes included AP-1, whose binding peaked about 6 hr after quinolinic acid administration, and E2F-1, which was only modestly and transiently elevated. In contrast, quinolinic acid lead to a reduction in OCT-1, beginning after 12 hr, and briefly in SP-1 binding. The NF kappa B, AP-1, and OCT-1 changes were attenuated both by the N-methyl-D-aspartate receptor antagonist MK-801 and the protein synthesis inhibitor cycloheximide. Moreover, quinolinic acid-induced internucleosomal DNA fragmentation and striatal cell death were significantly reduced by the intrastriatal administration of NF kappa B SN50, a cell-permeable recombinant peptide that blocks NF kappa B nuclear translocation. These results illustrate the complex temporal pattern of transcription factor change attending the apoptotic destruction produced in rat striatum by quinolinic acid. They further suggest that NF kappa B activation contributes to the excitotoxin-induced death of striatal neurons.


Mobile Mammography Screening as an Opportunity to Increase Access of Rural Women to Breast Cancer Research Studies.
Rural women are underrepresented in cancer research. We hypothesized that providing access to a research study to rural, medically underserved women who were receiving their breast cancer screening using a mobile mammography unit would increase the representation of rural women in a cancer cohort study. This study is a cross-sectional study using a cohort of women who have been recruited to a breast cancer study in Arkansas. Recruiters accompanied a mobile mammography unit, the MammoVan, to implement a novel method for reaching and recruiting underrepresented rural Arkansas women into the study. Participants include 5850 women recruited from 2010 through 2012 as part of the Arkansas Rural Community Health (ARCH) study. Participants recruited during their mammography screening on the MammoVan tended to be more rural, less educated, and more likely to be non-Hispanic than those recruited in other venues. A significant difference was not noted for race or age. Collaboration with the MammoVan greatly aided the recruitment of rural participants. These strategies can facilitate the representation of this historically underserved and understudied rural population in future research studies.


Relationships between vascularization and proliferation in invasive breast cancer.
Studies on relationships between angiogenesis and tumour cell proliferation have provided conflicting results. This study has therefore investigated the relationships between the number and location of fully automatically identified CD31-positive microvessels and interactively segmented mitoses and necrotic compartments by image processing. These features were studied in ten invasive breast cancers, in the 'hot spots' and in whole tumour sections. Microvessel and mitosis hot spots were topographically close or overlapping and were always located at the periphery of the tumour. The numbers of mitoses and microvessels per mm(2) in the hot spot were strongly correlated with the respective numbers in the whole tumour section, as well as mutually. The ratio of mitoses in the hot spot to the whole tumour section was significantly higher than the corresponding microvessel ratio. Mitoses were preferentially located at a distance of 50-150 microm from microvessels. No significant difference was found between the average distance between mitoses and microvessels in the whole tumour sections and in the hot spot (79 vs. 72 microm), although considerable inter-tumour differences were found (hot spot 43-101 microm, tumour 47-111 microm). The presence of necrotic areas correlated with the number of mitoses per mm(2) and necrosis was in general observed at a distance of more than 150 microm from the microvessels, suggesting that necrotic areas have outgrown their vascular system. These results indicate the usefulness of image processing of whole tumour sections for the identification of proliferation and vascularization hot spots, which are strong prognostic factors in breast cancer. The results also support a close relationship between tumour necrosis and microvessels.


Infected urocolpos and generalized peritonitis secondary to labia minora adhesions.
Labia minora adhesions (LMA) are a common finding in young girls. Usually, this condition is asymptomatic and spontaneously disappears during adolescence. We report on a case revealed by infected urocolpos and peritonitis and whose treatment finally required surgical reduction labioplasty. A 9-year-old girl presented with a 2-day history of abdominal pain and fever. Urinary continence had never been obtained, with diurnal leaks. Physical examination showed signs of peritoneal irritation and a subtotal vulvar obstruction due to LMA. At surgery, after LMA lysis, a large amount of cloudy urine-like fluid emptied under pressure from the vagina. Laparoscopy showed generalized peritonitis without any intraabdominal cause. The same Escherichia coli was identified in the infected urocolpos and the abdominal fluid. Postoperative course was uneventful. Because of recurrent LMA, the patient underwent several courses of local estrogen therapy. Labia minora hypertrophy with LMA developed 2 years after peritonitis, requiring surgical reduction labioplasty. We used a new technique with interposition of skin flaps. The girl is now well, without LMA or infection, 4 years after labioplasty. Although rare, subtotal vulvar obstruction because of LMA may lead to infected hydrocolpos and peritonitis. Recurrent LMA may necessitate surgical labioplasty.


An update on mortality and morbidity in patients with very low postoperative hemoglobin levels who decline blood transfusion (CME).
Severely anemic patients for whom blood transfusion is not an option provide highly valuable information on risks of anemia and alternative management strategies. This is a retrospective study of consecutive patients at least 18 years old who could not be transfused, had surgery at a patient blood management center between 2003 and 2012, and had at least one hemoglobin (Hb) measurement of not more than 8 g/dL during the postoperative period. The primary outcome measure was mortality, occurring anytime in the period after the surgery until discharge or 30th day after the surgery, whichever was sooner. Postoperative morbidities included the occurrence of any of sepsis, pneumonia, myocardial infarction, deep wound infection, congestive heart failure, and arrhythmia. A total of 293 patients (including 288 who self-identified as "Jehovah's Witness") were eligible and enrolled. The mean ± SD age of the patients was 61.5 ± 16.9 years and 74.1% were female. Overall mortality rate was 8.2% (95% confidence interval [CI], 5%-11.3%). Unadjusted odds ratio (OR) of death per each 1 g/dL decrease in the nadir postoperative Hb was 2.04 (95% CI, 1.52-2.74); OR of death after adjustment for other significant factors (urgency, American Society of Anesthesiology score, and age) was 1.82 (95% CI, 1.27-2.59). Our study confirms the previously reported low risk of mortality in upper nadir Hb ranges of 7 to 8 g/dL and much higher risk in lower ranges, albeit the number of patients reaching extremely low Hb levels were lower than previous report, possibly suggestive of improved management strategy of these patients.


How segmental duplications shape our genome: recent evolution of ABCC6 and PKD1 Mendelian disease genes.
The completion of the Human Genome Project has brought the understanding that our genome contains an unexpectedly large proportion of segmental duplications. This poses the challenge of elucidating the consequences of recent duplications on physiology. We have conducted an in-depth study of a subset of segmental duplications on chromosome 16. We focused on PKD1 and ABCC6 duplications because mutations affecting these genes are responsible for the Mendelian disorders autosomal dominant polycystic kidney disease and pseudoxanthoma elasticum, respectively. We establish that duplications of PKD1 and ABCC6 are associated to low-copy repeat 16a and show that such duplications have occurred several times independently in different primate species. We demonstrate that partial duplication of PKD1 and ABCC6 has numerous consequences: the pseudogenes give rise to new transcripts and mediate gene conversion, which not only results in disease-causing mutations but also serves as a reservoir for sequence variation. The duplicated segments are also involved in submicroscopic and microscopic genomic rearrangements, contributing to structural variation in human and chromosomal break points in the gibbon. In conclusion, our data shed light on the recent and ongoing evolution of chromosome 16 mediated by segmental duplication and deepen our understanding of the history of two Mendelian disorder genes.


Cascaded Thermodynamic and Environmental Analyses of Energy Generation Modalities of a High-Performance Building Based on Real-Time Measurements.
This study presents cascaded thermodynamic and environmental analyses of a high-performance academic building. Five different energy efficiency measures and operation scenarios are evaluated based on the actual measurements starting from the initial design concept. The study is to emphasize that by performing dynamical energy, exergy, exergoeconomic, and environmental analyses with increasing complexity, a better picture of building performance indicators can be obtained for both the building owners and users, helping them to decide on different investment strategies. As the first improvement, the original design is modified by the addition of a ground-air heat exchanger for pre-conditioning the incoming air to heat the ground floors. The installation of roof-top PV panels to use solar energy is considered as the third case, and the use of a trigeneration system as an energy source instead of traditional boiler systems is considered as the fourth case. The last case is the integration of all these three alternative energy modalities for the building. It is determined that the use of a trigeneration system provides a better outcome than the other scenarios for decreased energy demand, for cost reduction, and for the improved exergy efficiency and sustainability index values relative to the original baseline design scenario. Yet, an integrated approach combining all these energy generation modalities provide the best return of investment.


Use of Epstein-Barr virus-transformed B cell lines for the generation of immunoglobulin-producing human B cell hybridomas.
HGPRTase-deficient EBV-transformed B cell lines were shown to be effective fusion partners with mitogen-activated human B cells for the construction of Ig-producing human B cell hybridomas. In a series of experiments using these lines and B cells from several tissue sources, approximatley 20% of the cultures plated were consistently positive for growth after hypoxanthine-aminopterin-thymidine selection and approximatley 30% of these synthesized significant new Ig. A marked increase in Ig secretion was observed after hybridization, which was due to new Ig; Ig from the parental lime was shown to disappear in several instances. Special analyses were carried out on a human hybridoma secreting antibody specific for tetanus toxoid and tetanus toxin and stable subclones were derived. These studies suggest that EBV-transformed lines will prove useful in human hybridization studies, thus making a large library of B cell lines available for the generation of human monoclonal antibodies.


[Women and post-natal care: analysis of a rooming-in facility.]
The practice of rooming-in (R-in) is widely but heterogeneously applied in Italian hospitals and consists in keeping babies in the same room with their mothers from the moment of birth through to the discharge. Explore the perception and use of R-in among Italian mothers who gave birth in a context where R-in is offered 24h. We aimed at comparing differences among women who consistently use the R-in practice to those who do not. Sixty-five women in good health (average age=33.8 years old; sd=4.2) were assigned to two different groups: mothers who always kept their baby in the room (n=35) and mothers who referred their babies more than once to the nursery care (n=30). Women completed a questionnaire about their pregnancy, delivery, hospitalization, and R-in. Medical records were also collected. The two groups were compared with regard to medical/obstetrician/socio-demographic factors, as well as their perception of R-in. We observed no differences in terms of maternal age, education, and parity among the two groups. Women who had undergone Caesarean section were more likely to make use of the nursery care up to 72 hours after delivery (χ2=11,11, p<,001). A considerable proportion of the study population (42%) considered the level of information received about R-in unsatisfying; this percentage is higher (55%) among women who referred to the nursery. Women differ in the degree of R-in application use depending on the type of delivery and the subsequent maternal physical condition. It is important to support women by offering them a flexible service that accounts for their possible physical difficulties. Besides, it is crucial to provide an adequate level of information about R-in, so that mothers can feel in control of the situation, especially if they underwent Caesarean section.


[Disorders of the hemostasis system in late pregnancy toxemia and their correction].
The coagulation system has been examined in 30 women with late toxemia before and after heparin and trental therapy. Pretreatment fibrinogen levels were elevated and antithrombin III levels were reduced. A hemolysate-calcium mixture showed an increase in coagulation at 8 and 10 minutes of incubation. Staphylococcus agglutination and ethanol tests revealed a significant increase in early PDF and other constituents of the fibrinogen pool. These changes were indicative of prolonged-type disseminated intravascular coagulation in late toxemia. Comprehensive therapy comprising heparin and trental leads to a significant reduction in fibrinogen concentration, alleviation of the hypercoagulatory shift and increase in antithrombin III level. Early PDF and fibrin monomer complexes showed a decline. In addition, the therapy had beneficial effects on the maternal and fetal clinical status.


The role of networks and artificial intelligence in nanotechnology design and analysis.
Techniques with their origins in artificial intelligence have had a great impact on many areas of biomedicine. Expert-based systems have been used to develop computer-assisted decision aids. Neural networks have been used extensively in disease classification and more recently in many bioinformatics applications including genomics and drug design. Network theory in general has proved useful in modeling all aspects of biomedicine from healthcare organizational structure to biochemical pathways. These methods show promise in applications involving nanotechnology both in the design phase and in interpretation of system functioning.


Large sex differences in chicken behavior and brain gene expression coincide with few differences in promoter DNA-methylation.
While behavioral sex differences have repeatedly been reported across taxa, the underlying epigenetic mechanisms in the brain are mostly lacking. Birds have previously shown to have only limited dosage compensation, leading to high sex bias of Z-chromosome gene expression. In chickens, a male hyper-methylated region (MHM) on the Z-chromosome has been associated with a local type of dosage compensation, but a more detailed characterization of the avian methylome is limiting our interpretations. Here we report an analysis of genome wide sex differences in promoter DNA-methylation and gene expression in the brain of three weeks old chickens, and associated sex differences in behavior of Red Junglefowl (ancestor of domestic chickens). Combining DNA-methylation tiling arrays with gene expression microarrays we show that a specific locus of the MHM region, together with the promoter for the zinc finger RNA binding protein (ZFR) gene on chromosome 1, is strongly associated with sex dimorphism in gene expression. Except for this, we found few differences in promoter DNA-methylation, even though hundreds of genes were robustly differentially expressed across distantly related breeds. Several of the differentially expressed genes are known to affect behavior, and as suggested from their functional annotation, we found that female Red Junglefowl are more explorative and fearful in a range of tests performed throughout their lives. This paper identifies new sites and, with increased resolution, confirms known sites where DNA-methylation seems to affect sexually dimorphic gene expression, but the general lack of this association is noticeable and strengthens the view that birds do not have dosage compensation.


Lengthening of the duration of xylogenesis engenders disproportionate increases in xylem production.
In cold climates, the expected global warming will lead to earlier cambial resumptions in spring, with a resultant lengthening of the growing season but unknown consequences on forest productivity. The phenological traits of cambium activity and xylem formation were analyzed at a short time scale along a thermal gradient represented by an alti-latitudinal range from the 48th to 53rd parallels and covering the whole closed black-spruce [Picea mariana (Mill.) BSP] forest in Quebec, Canada. A hypothesis was tested that warmer temperatures influence cambium phenology, allowing longer duration and higher intensity of growth, and resulting in proportionally increased xylem production. From April to October 2012, cell division in cambium and post-cambial differentiation of xylem were observed on anatomical sections obtained from microcores collected weekly from the stem of fifty trees. The southern and warmer site was characterized by the highest radial growth, which corresponded to both the highest rates and longest durations of cell production. The differences in terms of xylem phenology and growth were marginal between the other sites. Xylem growth was positively correlated with rate and duration of cell production, with the latter explaining most variability in growth. Within the range analyzed, the relationship between temperature and most phenological phases of xylogenesis was linear. On the contrary, temperature was related with cell production according to an exponential pattern. Periods of xylogenesis of 14 days longer (+13.1%) corresponded to a massive increase in cell production (33 cells, +109%). This disproportionate change occurred at a May-September average temperature of ca. 14 °C and a snow-free period of 210-235 days. At the lower boundary of the distribution of black spruce, small environmental changes allowing marginal lengthening of the period of cell division could potentially lead to disproportionate increases in xylem cell production, with substantial consequences for the productivity of this boreal species.


How do novices learn physical examination skills? A systematic review of the literature.
Physical Examination (PE) skills are vital for patient care, and many medical students receive their first introduction to them in their pre-clinical years. A substantial amount of curriculum time is devoted to teaching these skills in most schools. Little is known about the best way to introduce PE skills to novice learners. Our objective was to conduct a systematic review of how medical students are first taught PE skills and the evidence supporting these strategies. We searched ERIC, SCOPUS, MEDLINE, PubMed and EMBASE for descriptions of complete PE curricula for novice learners. Inclusion criteria were: (1) English language; (2) subjects were enrolled in medical school and were in the preclinical portion of their training; (3) description of a method to teach physical examination skills for the first time; (4) description of the study population; (5) Description of a complete PE curriculum. We used the Medical Education Research Study Quality Instrument (MERSQI) score to evaluate the quality of evidence provided. Our search returned 5,418 articles; 32 articles met our inclusion criteria. Two main types of curricula were reported: comprehensive 'head-to-toe' PE curricula (18%) and organ system-based curricula (41%). No studies compared these directly, and only two evaluated trainees' clinical performance. The rest of the articles described interventions used across curricula (41%). Median MERSQI score was 10.1 Interquartile range 8.1-12.4. We found evidence for the use of non-faculty teaching associates, technology-enhanced PE education, and the addition of clinical exposure to formal PE teaching. The current literature on teaching PE is focused on describing innovations to head-to-toe and organ system-based curricula rather than their relative effectiveness, and is further limited by its reliance on short-term outcomes. The optimal strategy for novice PE instruction remains unknown.


[Intensive antibacterial therapy of multiple trauma].
Septic complications are considered among the main causes of death in patients with multiple injuries. In intensive care units their incidence is 33% of all cases. A total of 655 samples of sputum, blood, and urine from 224 patients were examined. The principal agents of septic infections were gram negative aerobic bacilli Pseudomonas aeruginosa and Proteus. The spectrum of activity of penicillins and aminoglycosides has appreciably changed. A decrease of contamination may be attained by arranging a special "cleansing" intensive care unit with continuous exchange of air free of bacteria, a regimen of an operative room, and use of solely cephalosporines combined with fluoroquinolones.


Efficient design of synthetic gene circuits under cell-to-cell variability.
Synthetic biologists use and combine diverse biological parts to build systems such as genetic circuits that perform desirable functions in, for example, biomedical or industrial applications. Computer-aided design methods have been developed to help choose appropriate network structures and biological parts for a given design objective. However, they almost always model the behavior of the network in an average cell, despite pervasive cell-to-cell variability. Here, we present a computational framework and an efficient algorithm to guide the design of synthetic biological circuits while accounting for cell-to-cell variability explicitly. Our design method integrates a Non-linear Mixed-Effects (NLME) framework into a Markov Chain Monte-Carlo (MCMC) algorithm for design based on ordinary differential equation (ODE) models. The analysis of a recently developed transcriptional controller demonstrates first insights into design guidelines when trying to achieve reliable performance under cell-to-cell variability. We anticipate that our method not only facilitates the rational design of synthetic networks under cell-to-cell variability, but also enables novel applications by supporting design objectives that specify the desired behavior of cell populations.


Effects of aerobic interval training on arterial stiffness and microvascular function in patients with metabolic syndrome.
The authors determined the effect of high-intensity aerobic interval training on arterial stiffness and microvascular dysfunction in patients with metabolic syndrome with hypertension. Applanation tonometry was used to measure arterial stiffness and laser Doppler flowmetry to assess microvascular dysfunction before and after 6 months of stationary cycling (training group; n = 23) in comparison to a group that remained sedentary (control group; n = 23). While no variable improved in controls, hypertension fell from 79% (59%-91%) to 41% (24%-61%) in the training group, resulting in lower systolic and diastolic pressures than controls (-12 ± 3 and -6 ± 2 mm Hg, P < .008). Arterial stiffness declined (-17% augmentation index, P = .048) and reactive hyperemia increased (20%, P = .028) posttreatment in the training group vs controls. Blood constituents associated with arterial stiffness and a prothrombotic state (high-sensitivity C-reactive protein, fibrinogen, platelets, and erythrocytes) remained unchanged in the training and control groups. In summary, 6 months of an intense aerobic exercise program reduced both arterial stiffness and microvascular dysfunction in patients with metabolic syndrome despite unchanged blood-borne cardiovascular risk factors. Training lowers blood flow resistance in central and peripheral vascular beds in a coordinated fashion, resulting in clinically relevant reductions in hypertension.


Outcomes of the IMMray PanCan-d Test in High-Risk Individuals Undergoing Pancreatic Surveillance: Pragmatic Data and Lessons Learned.
An effective blood-based test for pancreatic cancer (PC) screening has remained elusive. The IMMray PanCan-d is the first commercially available blood-based test specifically designed for early detection of PC; however, outcomes from its use in clinical practice have not been reported. We performed a blinded spike-in study of 100 individuals who had an IMMray PanCan-d test, including 94 high-risk individuals (HRIs) undergoing PC surveillance and six individuals with known PC. Specimens were processed blindly following the commercial laboratory's standardized operating procedure. Positive predictive value (PPV) and negative predictive value (NPV) were calculated. Cohort characteristics included a median age of 63 (IQR, 55-70) years, 57% female, 96% non-Hispanic White, 57% with a pathogenic variant in a PC risk gene (BRCA2 most commonly-18%), and 83% with a family history of PC. Among IMMray PanCan-d results from 94 HRIs undergoing PC surveillance, there was one positive (1%), seven borderlines (7%), 73 negatives (78%), and 13 tests not performed because of low CA19-9 expression (14%). No PC was diagnosed among these HRIs; however, there were two sub-cm pancreatic neuroendocrine tumors, seven clinically diagnosed side branch intraductal papillary mucinous neoplasms ≥1 cm, and a sub-cm solid mass with indeterminate cytology requiring close follow-up; all these individuals had negative IMMray PanCan-d tests. Of the six spiked-in PCs, four (67%) yielded a positive and two (33%) yielded a negative. With an estimated disease prevalence of 2%, the PPV and NPV are 52% and 99%, respectively, if borderline results are considered negative and 12% and 99%, respectively, if borderline tests are considered positive. In clinical practice, IMMray PanCan-d has a robust NPV; however, PPV is dramatically influenced by whether borderline results are characterized as a positive or negative result.


Silicone-rubber implant arthroplasty of the metacarpophalangeal joints for rheumatoid arthritis.
Forty-six patients with rheumatoid arthritis underwent metacarpophalangeal-joint arthroplasty of the index through little fingers on 210 joints in fifty-five hands using the Swanson-design silicone-rubber spacer. The patients were followed for two to eight years (average, five and one-quarter years). They were evaluated both preoperatively and postoperatively for range of motion, deformity, subjective sense, grip strength, and prehension. In the initial postoperative evaluation, the majority of patients expressed a strong subjective impression of improvement. Ulnar drift improved from the preoperative average of 25 degrees to less than 5 degrees. The preoperative average extension deficit decreased from 56 to 10 degrees, while the average range of motion increased from 17 to 51 degrees. In the long-term postoperative evaluation, the average ulnar drift had increased to 12 degrees, the average extension deficit had increased to 22 degrees, and the average range of motion had decreased to 39 degrees. Grip strength and prehension did not significantly improve at either evaluation. There were no fractures of the prosthesis and no patient had synovitis. We have found the procedure to be useful for the correction of deformity, increasing range of motion of the fingers, and improving the patient's sense of well-being.


Vitamin D supplementation and incident preeclampsia: A systematic review and meta-analysis of randomized clinical trials.
Maternal vitamin D deficiency has been associated with an increased risk for preeclampsia. Despite this, the current evidence regarding the efficacy of vitamin D supplementation in preventing preeclampsia is controversial. To assess the impact of vitamin D supplementation on the risk of preeclampsia, we performed a systematic review of the literature and a meta-analysis of the available randomized clinical trials (RCTs). The primary outcome was preeclampsia. Subgroup analyses were carried out considering the timing of the supplementation, type of intervention and the study design. Meta-regression analysis, including the amount of vitamin D and maternal age, were planned to explore heterogeneity (PROSPERO database registration number: CRD42019119207). Data were pooled from 27 RCTs comprising 59 arms, which included overall 4777 participants, of whom 2487 were in the vitamin D-treated arm and 2290 in the control arm. Vitamin D administration in pregnancy was associated with a reduced risk of preeclampsia (odd ratio [OR] 0.37, 95% confidence interval [CI]: 0.26, 0.52; I2 = 0%). If the vitamin D supplementation was started up to 20 weeks' gestation, the odds was a little lower (OR 0.35, 95% CI: 0.24, 0.50, p < 0.001). The effect was largely independent of the supplementation cessation (until delivery or not), type of intervention (vitamin D alone or in association with calcium), and study design. Increasing dose of vitamin D was associated with reduced incidence of preeclampsia (slope of log OR: -1.1, 95% CI: -1.73, -0.46; p < 0.001). Results suggest that vitamin D supplementation may be useful in preventing preeclampsia. These data are especially useful for health-care providers who engage in the management of pregnant women at risk for preeclampsia. Our findings are a call for action to definitively address vitamin D supplementation as a possible intervention strategy in preventing preeclampsia in pregnancy.


Pretreatment of liver grafts in vivo by γ-aminobutyric acid receptor regulation reduces cold ischemia/warm reperfusion injury in rat.
Gamma-aminobutyric acid (GABA) is found throughout the body. The regulation of GABA receptor (GABAR) reduces oxidative stress (OS). Ischemia/reperfusion injury after orthotopic liver transplantation (OLT) causes OS-induced graft damage. The effects of GABAR regulation in donors in vivo were investigated. Donor rats received saline, a GABAR agonist or GABAR antagonist 4 h before surgery. Recipient rats were divided into four groups according to the donor treatments: laparotomy, OLT with saline, OLT with GABAR agonist and OLT with GABAR antagonist. Histopathological, biochemical and immunohistological examinations were performed at 6, 12 and 24 h after OLT. Protein assays were performed at 6 h after OLT. The 4-hydroxynonenal (4-HNE), ataxia-telangiectasia mutated kinase (ATM), phosphorylated histone H2AX (gammaH2AX), phosphatidylinositol-3 kinase (PI3K), Akt and superoxide dismutase (SOD) were assessed by western blot analysis. In the univariate analysis, histopathological and biochemical profiles verified that the GABAR agonist reduced graft damage. Immunohistology revealed that the GABAR agonist prevented the induction of apoptosis. Measurement of 4-4-HNE levels confirmed OS-induced damage after OLT, and the GABAR agonist improved this damage. In the gammaH2AX, PI3K, Akt and antioxidant enzymes (SODs), ATM and H2AX were greatly increased after OLT, and were reduced by the GABAR agonist. In the multivariate analyses between multiple groups, histopathological assessment, aspartate aminotransferase level, immunohistological examinations for apoptotic induction and gammaH2AX showed statistical differences. A specific agonist demonstrated regulation of GABAR in vivo in the liver. This activation in vivo reduced OS after OLT via the ATM/H2AX pathway.


Ultrasensitive Ultrasound Microvessel Imaging for Characterizing Benign and Malignant Breast Tumors.
Tumor angiogenesis plays an important role during breast tumor growth. However, conventional Doppler has limited sensitivity to detect small blood vessels, resulting in a large overlap of Doppler features between benign and malignant tumors. An ultrasensitive ultrasound microvessel imaging (UMI) technique was recently developed. To evaluate the performance of UMI, we studied 44 patients with 51 breast masses. Tumor pathology served as the gold standard: 28 malignancies and 23 benignities. UMI provided a significant improvement in depicting smaller vessels compared with conventional Doppler. The microvessel morphologies observed on UMI were associated with tumor benign/malignant classification. The diagnostic accuracy of correct Breast Imaging Reporting and Data System (BI-RADS) classification rate (BI-RADS ≥4a: test positive; BI-RADS ≤3: test negative) as a fraction of total mass population was improved by 16% after combining conventional ultrasound with UMI compared with using conventional ultrasound alone. This improvement indicates the potential of UMI in reducing unnecessary benign biopsies and avoiding missed malignant biopsies.