Fusion of two distinct peptide exosite inhibitors of Factor VIIa.
Highly potent bifunctional inhibitors of Factor VIIa (FVIIa) were generated by linking two distinct peptides, recently shown to bind to two discrete exosites on the FVIIa protease domain [Dennis, Eigenbrot, Skelton, Ultsch, Santell, Dwyer, O'Connell and Lazarus (2000) Nature (London) 404, 465-470; Dennis, Roberge, Quan and Lazarus (2001) Biochemistry 40, 9513-9521; Roberge, Santell, Dennis, Eigenbrot, Dwyer and Lazarus (2001) Biochemistry 40, 9522-9531]. Fusion peptides consisting of an N-terminal A-series peptide followed by flexible linkers, an E-series peptide, and the Z-domain of protein A were expressed in Escherichia coli and purified using IgG-Sepharose affinity chromatography. The fusion peptides were potent anticoagulants and had steep concentration dependence curves in tissue factor-dependent prothrombin time (PT) assays in comparison to the individual peptides or their noncovalent combination. This phenomenon was dependent on the length of the linker joining the A- and E-peptides. The fusion of the peptides increased the extent of inhibition of Factor X (FX) activation to 100% at saturating peptide concentrations, but did not improve the binding affinity for Factor VIIa (FVIIa) at the A- and E- binding sites or the IC(50) for the inhibition of FX activation. Differences between the peptides in the PT fold prolongation in normal and FVII-deficient plasma, in conjunction with the inhibition of (125)I-FVII activation, suggest that the enhanced effects of the fusion peptides involve the inhibition of FVII autoactivation.


No contribution of the ABCB11 p.444A polymorphism in Japanese patients with drug-induced cholestasis.
European studies have revealed that the ABCB11 c.1331T>C (V444A) polymorphism (rs2287622) C-allele frequency is higher among patients with drug-induced cholestasis. Given the low incidence of this disease, however, this association has not been sufficiently elucidated. We aimed to investigate the significance of this polymorphism in Japanese patients. We determined ABCB11 V444A polymorphism frequencies and HLA genotypes in two independent drug-induced cholestasis cohorts. Expression and taurocholate transport activity of proteins from 444A variants were analyzed using Madin-Darby canine kidney II cells. In cohort 1 (n = 40), the V444A polymorphism C-allele frequency (66%) was lower than that in controls (n = 190, 78%), but this difference was not significant (P = 0.09). In cohort 2 (n = 119), comprising patients with cholestatic (n = 19), hepatocellular (n = 74), and mixed (n = 26) liver injuries, the C-allele frequency was lower among patients with cholestatic liver injury (68%) than among those with hepatocellular (75%) or mixed liver injury (83%), although this difference was not significant. In cohort 1, HLA-A*0201 was observed more frequently in patients (22%) than in controls [11%; P = 0.003; odds ratio, 2.4 (95% confidence interval, 1.4-4.0)]. Taurocholate transport activity of 444A-encoded protein was significantly lower than that of 444V-encoded protein (81% of 444V, P < 0.05) because of the reduced protein stability. In conclusion, ABCB11 444A had slightly reduced transport activity, but it did not contribute to the occurrence of drug-induced cholestasis in Japanese patients. Therefore, genetic susceptibility to acquired cholestasis may differ considerably by ethnicity.


Multilocus sequence type analysis reveals both clonality and recombination in populations of Candida glabrata bloodstream isolates from U.S. surveillance studies.
The human commensal yeast Candida glabrata is becoming increasingly important as an agent of nosocomial bloodstream infection. However, relatively little is known concerning the genetics and population structure of this species. We have analyzed 230 incident bloodstream isolates from previous and current population-based surveillance studies by using multilocus sequence typing (MLST). Our results show that in the U.S. cities of Atlanta, GA; Baltimore, MD; and San Francisco, CA during three time periods spanning 1992 to 2009, five populations of C. glabrata bloodstream isolates are defined by a relatively small number of sequence types. There is little genetic differentiation in the different C. glabrata populations. We also show that there has been a significant temporal shift in the prevalence of one major subtype in Atlanta. Our results support the concept that both recombination and clonality play a role in the population structure of this species.


RET mutational spectrum in Hirschsprung disease: evaluation of 601 Chinese patients.
Rare (RVs) and common variants of the RET gene contribute to Hirschsprung disease (HSCR; congenital aganglionosis). While RET common variants are strongly associated with the commonest manifestation of the disease (males; short-segment aganglionosis; sporadic), rare coding sequence (CDS) variants are more frequently found in the lesser common and more severe forms of the disease (females; long/total colonic aganglionosis; familial).Here we present the screening for RVs in the RET CDS and intron/exon boundaries of 601 Chinese HSCR patients, the largest number of patients ever reported. We identified 61 different heterozygous RVs (50 novel) distributed among 100 patients (16.64%). Those include 14 silent, 29 missense, 5 nonsense, 4 frame-shifts, and one in-frame amino-acid deletion in the CDS, two splice-site deletions, 4 nucleotide substitutions and a 22-bp deletion in the intron/exon boundaries and 1 single-nucleotide substitution in the 5' untranslated region. Exonic variants were mainly clustered in RET the extracellular domain. RET RVs were more frequent among patients with the most severe phenotype (24% vs. 15% in short-HSCR). Phasing RVs with the RET HSCR-associated haplotype suggests that RVs do not underlie the undisputable association of RET common variants with HSCR. None of the variants were found in 250 Chinese controls.


MARTs and MARylation in the Cytosol: Biological Functions, Mechanisms of Action, and Therapeutic Potential.
Mono(ADP-ribosyl)ation (MARylation) is a regulatory post-translational modification of proteins that controls their functions through a variety of mechanisms. MARylation is catalyzed by mono(ADP-ribosyl) transferase (MART) enzymes, a subclass of the poly(ADP-ribosyl) polymerase (PARP) family of enzymes. Although the role of PARPs and poly(ADP-ribosyl)ation (PARylation) in cellular pathways, such as DNA repair and transcription, is well studied, the role of MARylation and MARTs (i.e., the PARP 'monoenzymes') are not well understood. Moreover, compared to PARPs, the development of MART-targeted therapeutics is in its infancy. Recent studies are beginning to shed light on the structural features, catalytic targets, and biological functions of MARTs. The development of new technologies to study MARTs have uncovered essential roles for these enzymes in the regulation of cellular processes, such as RNA metabolism, cellular transport, focal adhesion, and stress responses. These insights have increased our understanding of the biological functions of MARTs in cancers, neuronal development, and immune responses. Furthermore, several novel inhibitors of MARTs have been developed and are nearing clinical utility. In this review, we summarize the biological functions and molecular mechanisms of MARTs and MARylation, as well as recent advances in technology that have enabled detection and inhibition of their activity. We emphasize PARP-7, which is at the forefront of the MART subfamily with respect to understanding its biological roles and the development of therapeutically useful inhibitors. Collectively, the available studies reveal a growing understanding of the biochemistry, chemical biology, physiology, and pathology of MARTs.


Snus use, smoking and survival among prostate cancer patients.
Smoking is associated with prostate cancer mortality. The Scandinavian smokeless tobacco product snus is a source of nicotine but not the combustion products of smoke and has not been studied with respect to prostate cancer survival. The study is nested among 9,582 men with incident prostate cancer within a prospective cohort of 336,381 Swedish construction workers. Information on tobacco use was collected at study entry between 1971 and 1992, and categorized into (i) never users of any tobacco, (ii) exclusive snus: ever users of snus only, (iii) exclusive smokers: ever smokers (cigarette, cigar and/or pipe) only and (iv) ever users of both snus and smoking. Hazard ratios for prostate cancer-specific and total mortality for smoking and snus use based on Cox proportional hazards models adjusted for age, calendar period at diagnosis and body mass index at baseline. During 36 years of follow-up, 4,758 patients died-2,489 due to prostate cancer. Compared to never users of tobacco, exclusive smokers were at increased risk of prostate cancer mortality (HR 1.15, 95% CI: 1.05-1.27) and total mortality (HR 1.17, 95% CI: 1.09-1.26). Exclusive snus users also had increased risks for prostate cancer mortality (HR 1.24, 95% CI: 1.03-1.49) and total mortality (HR 1.19, 95% CI: 1.04-1.37). Among men diagnosed with nonmetastatic disease, the HR for prostate cancer death among exclusive snus users was 3.17 (95% CI: 1.66-6.06). The study is limited by a single assessment of tobacco use prior to diagnosis. Snus use was associated with increased risks of prostate cancer and total mortality among prostate cancer patients. This suggests that tobacco-related components such as nicotine or tobacco-specific carcinogens may promote cancer progression independent of tobacco's combustion products.


[Fetal atrioventricular septal defect associated with Patau and Edwards syndromes, as well as trisomy 22].
The atrioventricular septal defect is usually associated with trisomy 21 and it may be observed in the heterotaxia syndromes. Atrioventricular septal defect may be associated with 8p deletion. There are reported cases of familial atrioventricular septal defect. Atrioventicular septal defect is rarely associated with other chromosomal abnormalities. We are reporting three unusual cases of atrioventricular septal defect that were associated with trisomy 13, 18 and 22. This association may be due to effect of genetic loci on the 13, 18 and 22 chromosome which could play the role in the development and fusion of endocardial cushion and atrioventricular septal defect.


[Professional strategy and institutional isomorphism: the dental health insurance industry in Brazil].
This article analyzes the organizational model of the dental health industry. The main organizational leaders in this industry are the professional cooperatives and group dental insurance companies. The theoretical basis of the article is the organizational theory developed by Di Maggio and Powell. The dental health industry consists of a great number of small and very dynamic companies, however an expressive part of clients and profit are concentrated in a few large companies. The results show that the industry has expanded the number of clients after the creation of the National Health Insurance Agency. The regulation regime has forced institutional changes in the firms with regard to the market entry, permanence or exit patterns. There was no evidence that the regulatory rules have interfered with the development and financial conditions of the industry. The average profitability of the sector, especially among the group dental insurance companies, is extremely high.


Standardization of Small Animal Imaging-Current Status and Future Prospects.
The benefit of small animal imaging is directly linked to the validity and reliability of the collected data. If the data (regardless of the modality used) are not reproducible and/or reliable, then the outcome of the data is rather questionable. Therefore, standardization of the use of small animal imaging equipment, as well as of animal handling in general, is of paramount importance. In a recent paper, guidance for efficient small animal imaging quality control was offered and discussed, among others, the use of phantoms in setting up a quality control program (Osborne et al. 2016). The same phantoms can be used to standardize image quality parameters for multi-center studies or multi-scanners within center studies. In animal experiments, the additional complexity due to animal handling needs to be addressed to ensure standardized imaging procedures. In this review, we will address the current status of standardization in preclinical imaging, as well as potential benefits from increased levels of standardization.


Reduction of waste product excretion via nutrient control: Possible strategies for maximizing product and cell yields on serum in cultures of mammalian cells.
Mammalian cells grown in culture excrete lactic acid and ammonium ions in quantities that may limit growth and reduce product synthesis. Frequent replenishment of the culture medium is often necessary to prevent waste product accumulation which could inhibit cell growth. Since increased medium replenishment results in increased usage of animal serum, the most expensive raw material, excessive production of waste products lowers the cell and product yield on serum, and hence increases production costs. Strategies for reducing the production of lactic acid and ammonium bymammalian cells via controlled addition of glucose and glutamine will be demonstrated. Mathematical relations coupling ammonium and glutamine kinetics will be described. Additionally, a method for automatic on-line estimation of the cell concentration was developed. This method involves calculating the ATP production rate from the oxygen uptake rate and the lactic acid production rate. Automatic online estimation of the cell concentration is critical if nutrient levels in large-scale mammaliancell cultures are to be accurately maintained via process control.


Di-5-ANEQ(F)PTEA Offers Better Performance than Di-4-ANEQ(F)PTEA for In-Situ Cardiac Optical Mapping.
Cardiac optical mapping has traditionally been performed in ex-vivo, motion-arrested hearts. Recently, in-situ cardiac optical mapping has been made possible by both motion correction techniques and long-wavelength voltage sensitive dyes (VSDs). However, VSDs have been observed to wash out quickly from blood-perfused in-situ hearts. In this study, we evaluate the performance of a newly developed VSD, di-5-ANEQ(F)PTEA, relative to an earlier VSD, di-4-ANEQ(F)PTEA. We find that di-5-ANEQ(F)PTEA persists over 3 times longer, produces improved signal-to-noise ratio, and does not prolong loading unacceptably.Clinical Relevance-Optical mapping has provided many insights into cardiac arrhythmias, but has traditionally been limited to ex-vivo preparations. The present findings extend the utility of optical mapping in the more realistic in-vivo setting and may eventually enable its use in patients.


Immunological sterilization of male dogs by BCG.
The effect of a single intratesticular injection of Freund's complete adjuvant (FCA), incomplete Freund's adjuvant (IFA) and various components has been investigated in dogs. FCA caused severe granulomatous reaction with wide-spread degeneration and vacuolation of the tubules. IFA and Arlacel A induced oligo - but not azoospermia, which was of short duration (6 weeks). Bacillus Calmette Guerin (BCG) suspended in saline given with uniform distribution at doses of 10 units or higher was effective in inducing aspermatogenesis. The sperm count declined to near zero level within 3 to 6 weeks after injection of BCG, and sperm suppression was maintained for 6 to 11 months of the observation period. At moderate doses of 10 to 25 units partial to complete inhibition of spermatogenesis was seen in different tubules with leukocyte infiltration in the interstitium. The basement membrane of the tubules, the peritubular cells as well as the Leydig cells were intact. At high doses (75-110 units) and at sites where bacillary deposits were high even in moderate dose animals, there was massive leukocyte infiltration with nonspecific damage to the tubules. The animals, on low or high dose, retained libido and copulated with bitches in oestrus. The mating was, however, infertile. The treatment, especially with low and moderate doses, was largely free of significant side effects. At higher doses, scrotal pain and swelling lasting for 7-15 days was observed; a small decrease in the size of the testes was also noted.


[Pharmacokinetics and pharmacodynamics of morphinomimetics in the central nervous system].
The penetration of opioids into interstitial space of the brain is related to their physicochemical properties. All opioids are weak bases with different lipid solubilities; the octanol-water partition coefficient varies from 1.4 to 1700 for morphine and sufentanil, respectively. Lipid solubility is an essential factor for diffusion capacity across the capillary walls but the rate at which drug enters the brain by simple passive diffusion is more related to the diffusible fraction (unbound, unionized) for fentanyl and its derivatives. The relative potency of the analgesic effect is related to the characteristics of the opiate receptor interaction (KD, Ki). Sufentanil is the most potent of the opioids currently available for use in man, it has a very high binding affinity for the mu-opioid receptor. The duration of the pharmacological effect is determined by the time of dissociation of stereospecific drug-receptor binding and also by pharmacokinetic half-life; the recovery would be expected in consideration of the different terminal half-times.


Rediscovery of Bipalium admarginatum de Beauchamp, 1933 (Platyhelminthes, Tricladida, Geoplanidae) in Malaysia, with molecular characterisation including the mitogenome.
We present here the first observation of Bipalium admarginatum de Beauchamp, 1933 since its original description 90 years ago. Three specimens were found on Perhentian Kecil Island, off Terengganu State, Malaysia and photographed in the field, and two were collected. This report thus includes the first colour photographs published for this species, from a locality close to the type-locality, Tioman Island (which is ca. 200 km south of the locality in this study, on the east coast of Peninsula Malaysia). We describe the external morphology and colour pattern of the species, which correspond well to the original description, itself based only on two preserved specimens. We performed an in-depth molecular characterisation of the species, including its complete mitochondrial genome, the 18S sequence and elongation 1-alpha (EF1-α) sequence. In addition, EF1-α sequences were also retrieved for 5 additional geoplanid species. No tRNA-Thr could be detected in the mitogenome of B. admarginatum, a lack already reported in several species of geoplanids, but we found a 13 bp sequence that contains the anticodon loop and seems to be conserved among geoplanids and might thus possibly represent a non-canonical undetected tRNA. We discuss the difficulties encountered in trying to reconstruct the cluster of nuclear ribosomal genes, a problem already mentioned for other Triclads. Three phylogenies, based respectively on all mitochondrial proteins, 18S, and EF1-α, were computed; the position of B. admarginatum within the Bipaliinae was confirmed in each tree, as sister-group to various bipaliine species according to the sequences available for each tree. In the mitochondrial proteins tree, which had high support, B. admarginatum was sister to Bipalium kewense and Diversibipalium multilineatum.


Spontaneous metastasis of rat liver epithelial cells transformed with v-raf and v-raf/v-myc: association with different phenotypic properties.
Cloned v-raf, v-raf/v-myc, and spontaneously transformed rat liver epithelial (RLE) cell lines were examined for meastatic capability in nude mice, using the LacZ gene as a marker for quantitation of micrometastases. Six cloned lines (R3611-T lines) derived from nude mouse xenografts of the v-raf transformed R3611-3 cells displayed variable metastatic capabilities. Three of six subcutaneously inoculated R3611-TlacZ lines produced spontaneous lung metastasis in nude mice. One of the lines, R3611-T2lacZ was highly efficient at metastatic conversion and produced more lung colonies than a faster growing v-raf/v-myc-transformed RJ2-14lacZ line. The spontaneously transformed RLElacZ line (C4T) was nonmetastatic, although it produced larger subcutaneous tumors than the metastatic R3611-T2lacZ line. Metastatic conversion correlated with upregulation of urokinase-type plasminogen activator receptor RNA expression and downregulation of plasminogen activator inhibitor-1, collagen alpha1 (I), and cytokeratin 14 (K14) RNA expression. These findings indicate that proteolytic activities associated with plasminogen activation play a role in the metastatic development in this model. Decreased production of extracellular proteins and cytoskeletal changes associated with lack of K14 expression are also likely to have contributed to the metastatic conversion of the RLE transformants.


Regulation of Kv4.3 closed state inactivation and recovery by extracellular potassium and intracellular KChIP2b.
Mechanisms underlying Kv4 channel inactivation and recovery are presently unclear, although there is general consensus that the basic characteristics of these processes are not consistent with Shaker (Kv1) N- and P/C-type mechanisms. Kv4 channels also differ from Shaker in that they can undergo significant inactivation from pre-activated closed-states (closed-state inactivation, CSI), and that inactivation and recovery kinetics can be regulated by intracellular KChIP2 isoforms. To gain insight into the mechanisms regulating Kv4.3 CSI and recovery, we have analyzed the effects of increasing [K(+)](o) from 2 mM to 98 mM in the absence and in the presence of KChIP2b, the major KChIP2 isoform expressed in the mammalian ventricle. In the absence of KChIP2b, high [K(+)](o) promoted Kv4.3 inactivated closed-states and significantly slowed the kinetics of recovery from both macroscopic and closed-state inactivation. Coexpression of KChIP2b in 2 mM [K(+)](o) promoted non-inactivated closed-states and accelerated the kinetics of recovery from both macroscopic and CSI. In high [K(+)](o), KChIP2b eliminated or significantly reduced the slowing effects on recovery. Attenuation of CSI by the S4 charge-deletion mutant R302A, which produced significant stabilization of non-inactivated closed-states, effectively eliminated the opposing effects of high [K(+)](o) and KChIP2b on macroscopic recovery kinetics, confirming that these results were due to alterations of CSI. Elevated [K(+)](o) therefore slows Kv4.3 recovery by stabilizing inactivated closed-states, while KChIP2b accelerates recovery by destabilizing inactivated closed-states. Our results challenge underlying assumptions of presently popular Kv4 gating models and suggest that Kv4.3 possesses novel allosteric mechanisms, which are absent in Shaker, for coupling interactions between intracellular KChIP2b binding motifs and extracellular K(+)-sensitive regulatory sites.


A novel missense mutation (N258S) in the KCNQ2 gene in a Turkish family afflicted with benign familial neonatal convulsions (BFNC).
Benign familial neonatal convulsions (BFNC) is a rare monogenic subtype of idiopathic epilepsy exhibiting autosomal dominant mode of inheritance. The disease is caused by mutations in the two homologous genes KCNQ2 and KCNQ3 that encode the subunits of the voltage-gated potassium channel. Most KCNQ2 mutations are found in the pore region and the cytoplasmic C domain. These mutations are either deletions/insertions that result in frameshift or truncation of the protein product, splice-site variants or missense mutations. This study reveals a novel missense mutation (N258S) in the KCNQ2 gene between the S5 domain and the pore of the potassium channel in two BFNC patients in a Turkish family. The absence of the mutation both in the healthy members of the family and in a control group, and the lack of any other change in the KCNQ2 gene of the patients indicate that N258S substitution is a pathogenic mutation leading to epileptic seizures in this family.


A nuclear-targeted effector of Rhizophagus irregularis interferes with histone 2B mono-ubiquitination to promote arbuscular mycorrhisation.
Arguably, symbiotic arbuscular mycorrhizal (AM) fungi have the broadest host range of all fungi, being able to intracellularly colonise root cells in the vast majority of all land plants. This raises the question how AM fungi effectively deal with the immune systems of such a widely diverse range of plants. Here, we studied the role of a nuclear-localisation signal-containing effector from Rhizophagus irregularis, called Nuclear Localised Effector1 (RiNLE1), that is highly and specifically expressed in arbuscules. We showed that RiNLE1 is able to translocate to the host nucleus where it interacts with the plant core nucleosome protein histone 2B (H2B). RiNLE1 is able to impair the mono-ubiquitination of H2B, which results in the suppression of defence-related gene expression and enhanced colonisation levels. This study highlights a novel mechanism by which AM fungi can effectively control plant epigenetic modifications through direct interaction with a core nucleosome component. Homologues of RiNLE1 are found in a range of fungi that establish intimate interactions with plants, suggesting that this type of effector may be more widely recruited to manipulate host defence responses.


Predictors of loss of lung function in the elderly: the Cardiovascular Health Study.
Pulmonary function, as measured by spirometry (FEV1 or FVC), is an important independent predictor of morbidity and mortality in elderly persons. In this study we examined the predictors of longitudinal decline in lung function for participants of the Cardiovascular Health Study (CHS). The CHS was started in 1990 as a population-based observational study of cardiovascular disease in elderly persons. Spirometry testing was conducted at baseline, 4 and 7 yr later. The data were analyzed using a random effects model (REM) including an AR(1) error structure. There were 5,242 subjects (57.6% female, mean age 73 yr, 87.5% white and 12.5% African-American) with eligible FEV1 measures representing 89% of the baseline cohort. The REM results showed that African-Americans had significantly lower spirometry levels than whites but that their rate of decline with age was significantly less. Subjects reporting congestive heart failure (CHF), high systolic blood pressure (> 160 mm Hg), or taking beta-blockers had significantly lower spirometry levels; however, the effects of high blood pressure and taking beta-blockers diminished with increasing age. Chronic bronchitis, pneumonia, emphysema, and asthma were associated with reduced spirometry levels. The most notable finding of these analyses was that current smoking (especially for men) was associated with more rapid rates of decline in FVC and FEV1. African-Americans (especially women) had slower rates of decline in FEV1 than did whites. Although participants with current asthma had a mean 0.5 L lower FEV1 at their baseline examination, they did not subsequently experience more rapid declines in FEV1.


Swim bladder enhances lagenar sensitivity to sound pressure and higher frequencies in female plainfin midshipman (Porichthys notatus).
The plainfin midshipman fish (Porichthys notatus) is an established model for investigating acoustic communication because the reproductive success of this species is dependent on the production and reception of social acoustic signals. Previous work showed that female midshipman have swim bladders with rostral horn-like extensions that project close to the saccule and lagena, while nesting (type I) males lack such rostral swim bladder extensions. The relative close proximity of the swim bladder to the lagena should increase auditory sensitivity to sound pressure and higher frequencies. Here, we test the hypothesis that the swim bladder of female midshipman enhances lagenar sensitivity to sound pressure and higher frequencies. Evoked potentials were recorded from auditory hair cell receptors in the lagena in reproductive females with intact (control condition) and removed (treated condition) swim bladders while pure tone stimuli (85-1005 Hz) were presented by an underwater speaker. Females with intact swim bladders had auditory thresholds 3-6 dB lower than females without swim bladders over a range of frequencies from 85 to 405 Hz. At frequencies from 545 to 1005 Hz, only females with intact swim bladders had measurable auditory thresholds (150-153 dB re. 1 µPa). The higher percentage of evoked lagenar potentials recorded in control females at frequencies >505 Hz indicates that the swim bladder extends the bandwidth of detectable frequencies. These findings reveal that the swim bladders in female midshipman can enhance lagenar sensitivity to sound pressure and higher frequencies, which may be important for the detection of behaviorally relevant social signals.


Biochemical Characterization of Medaka (Oryzias latipes) Transglutaminases, OlTGK1 and OlTGK2, as Orthologues of Human Keratinocyte-Type Transglutaminase.
Calcium-dependent transglutaminases (TGs) are a family of enzymes that catalyze protein cross-linking and/or attachment of primary amines in a variety of organisms. Mammalian TGs are implicated in multiple biological events such as skin formation, blood coagulation, and extracellular matrix stabilization. Medaka (Oryzias latipes) has been used as a model fish to investigate the physiological functions of mammalian proteins. By analysis of the medaka genome, we found seven TGs orthologues, some of which apparently corresponded to the mammalian TG isozymes, TG1, TG2, and Factor XIII. All orthologues had preserved amino acid residues essential for enzymatic activity in their deduced primary structures. In this study, we analyzed biochemical properties of two orthologues (OlTGK1 and OlTGK2) of mammalian epithelium-specific TG (TG1) that are significantly expressed at the transcriptional level. Using purified recombinant proteins for OlTGK1 and OlTGK2, we characterized their catalytic reactions. Furthermore, immunohistochemical analyses of fish sections revealed higher expression in the pancreas (OTGK1), intervertebral disk (OlTGK2) and pharyngeal teeth (OlTGK2) as well as in the skin epidermis.


A hot hole-programmed and low-temperature-formed SONOS flash memory.
In this study, a high-performance TixZrySizO flash memory is demonstrated using a sol-gel spin-coating method and formed under a low annealing temperature. The high-efficiency charge storage layer is formed by depositing a well-mixed solution of titanium tetrachloride, silicon tetrachloride, and zirconium tetrachloride, followed by 60 s of annealing at 600°C. The flash memory exhibits a noteworthy hot hole trapping characteristic and excellent electrical properties regarding memory window, program/erase speeds, and charge retention. At only 6-V operation, the program/erase speeds can be as fast as 120:5.2 μs with a 2-V shift, and the memory window can be up to 8 V. The retention times are extrapolated to 106 s with only 5% (at 85°C) and 10% (at 125°C) charge loss. The barrier height of the TixZrySizO film is demonstrated to be 1.15 eV for hole trapping, through the extraction of the Poole-Frenkel current. The excellent performance of the memory is attributed to high trapping sites of the low-temperature-annealed, high-κ sol-gel film.


Infants with esophageal atresia and right aortic arch: Characteristics and outcomes from the Midwest Pediatric Surgery Consortium.
Right sided aortic arch (RAA) is a rare anatomic finding in infants with esophageal atresia with or without tracheoesophageal fistula (EA/TEF). In the presence of RAA, significant controversy exists regarding optimal side for thoracotomy in repair of the EA/TEF. The purpose of this study was to characterize the incidence, demographics, surgical approach, and outcomes of patients with RAA and EA/TEF. A multi-institutional, IRB approved, retrospective cohort study of infants with EA/TEF treated at 11 children's hospitals in the United States over a 5-year period (2009 to 2014) was performed. All patients had a minimum of one-year follow-up. In a cohort of 396 infants with esophageal atresia, 20 (5%) had RAA, with 18 having EA with a distal TEF and 2 with pure EA. Compared to infants with left sided arch (LAA), RAA infants had a lower median birth weight, (1.96 kg (IQR 1.54-2.65) vs. 2.57 kg (2.00-3.03), p = 0.01), earlier gestational age (34.5 weeks (IQR 32-37) vs. 37 weeks (35-39), p = 0.01), and a higher incidence of congenital heart disease (90% vs. 32%, p < 0.0001). The most common cardiac lesions in the RAA group were ventricular septal defect (7), tetralogy of Fallot (7) and vascular ring (5). Seventeen infants with RAA underwent successful EA repair, 12 (71%) via right thoracotomy and 5 (29%) through left thoracotomy. Anastomotic strictures trended toward a difference in RAA patients undergoing right thoracotomy for primary repair of their EA/TEF compared to left thoracotomy (50% vs. 0%, p = 0.1). Side of thoracotomy in RAA patients undergoing EA/TEF repair was not significantly associated with mortality, anastomotic leak, recurrent laryngeal nerve injury, recurrent fistula, or esophageal dehiscence (all p > 0.29). RAA in infants with EA/TEF is rare with an incidence of 5%. Compared to infants with EA/TEF and LAA, infants with EA/TEF and RAA are more severely ill with lower birth weight and higher rates of prematurity and complex congenital heart disease. In neonates with RAA, surgical repair of the EA/TEF is technically feasible via thoracotomy from either chest. A higher incidence of anastomotic strictures may occur with a right-sided approach. Level III.


Mechanism of action of the peptide antibiotic nisin in liposomes and cytochrome c oxidase-containing proteoliposomes.
The interaction of the peptide antibiotic nisin with liposomes has been studied. The effect of this interaction was analyzed on the membrane potential (inside negative) and the pH gradient (inside alkaline) in liposomes made from Escherichia coli phosphatidylethanolamine and egg phosphatidylcholine (9:1, wt/wt). The membrane potential and pH gradient were generated by artificial ion gradients or by the oxidation of ascorbate, N,N,N',N'-tetramethyl-p-phenylenediamine, and cytochrome c by the beef heart cytochrome c oxidase incorporated in the liposomal membranes. Nisin dissipated the membrane potential and the pH gradient in both types of liposomes and inhibited oxygen consumption by cytochrome c oxidase in proteoliposomes. The dissipation of the proton motive force in proteoliposomes was only to a minor extent due to a decrease of the oxidase activity by nisin. The results in these model systems show that a membrane potential and/or a pH gradient across the membrane enhances the activity of nisin. Nisin incorporates into the membrane and makes the membrane permeable for ions. As a result, both the membrane potential and pH gradient are dissipated. The activity of nisin was found to be influenced by the phospholipid composition of the liposomal membrane.


MicroRNA Regulation of Atherosclerosis.
Atherosclerosis and its attendant clinical complications, such as myocardial infarction, stroke, and peripheral artery disease, are the leading cause of morbidity and mortality in Western societies. In response to biochemical and biomechanical stimuli, atherosclerotic lesion formation occurs from the participation of a range of cell types, inflammatory mediators, and shear stress. Over the past decade, microRNAs (miRNAs) have emerged as evolutionarily conserved, noncoding small RNAs that serve as important regulators and fine-tuners of a range of pathophysiological cellular effects and molecular signaling pathways involved in atherosclerosis. Accumulating studies reveal the importance of miRNAs in regulating key signaling and lipid homeostasis pathways that alter the balance of atherosclerotic plaque progression and regression. In this review, we highlight current paradigms of miRNA-mediated effects in atherosclerosis progression and regression. We provide an update on the potential use of miRNAs diagnostically for detecting increasing severity of coronary disease and clinical events. Finally, we provide a perspective on therapeutic opportunities and challenges for miRNA delivery in the field.


Social cognition in Wilson's disease: A new phenotype?
Studies focusing on neuropsychological impairments in Wilson's disease (WD) have highlighted that patients showing neurological signs present significant deficits in a wide range of cognitive domains. Attentional and executive impairments have also been described in people with hepatic WD. However, social cognition abilities, i.e. cognitive processes required to perceive the emotions, intentions and dispositions of other people, have not been clearly investigated in WD. In this study we examined the social cognitive functioning in 19 patients with WD depending on their clinical status-Neurological versus Non-Neurological ("hepatic") forms-compared to 20 healthy controls. For the very first time, results highlighted that patients with WD had significant impairments in the three major components of social cognition: emotion recognition, Theory of Mind and attributional style. However, these deficits differ depending on the form of the disease: patients with neurological signs showed a wide range of deficits in the three components that were assessed-results notably revealed impairments in recognizing "fear", "anger", and "disgust", a significant Theory of Mind deficit and an "aggression bias"-whereas Non-Neurological patients only showed deficits on test assessing attributional bias, with a trend to react more "aggressively" to ambiguous social situations than healthy controls, as observed in Neurological WD patients, and a specific impairment in "anger" recognition. Our findings are discussed in the light of both neurocognitive impairments and brain damages, and especially those affecting the basal ganglia, as observed in people with WD.


Human activity classification with inertial sensors.
Monitoring human physical activity has become an important research area and is essential to evaluate the degree of functional performance and general level of activity of a person. The discrimination of daily living activities can be implemented with machine learning techniques. A public dataset provided during the European Symposium on Artificial Neural Networks 2013, with time and frequency domain features extracted from raw signals of the smartphone inertial sensors, was used to implement and evaluate an activity classifier. Using a decision tree classifier, an accuracy of 86% was achieved for the classification of walk, climb stairs, stand, sit, and lay down. The results obtained suggest that the smartphone's inertial sensors could be used for an accurate physical activity classification even with real-time requirements.


Simulations of the effect of an oxide on contact area measurements from conductive atomic force microscopy.
Nanoscale contact area in conductive atomic force microscopy can be determined by analyzing current flow using electron transport theories. However, it is recognized that native oxides on the conductive tip will reduce current flow, thus degrading the accuracy of the measured contact area. To quantify the adverse effect of an oxide on contact area measurements, we use molecular dynamics simulations of an oxide-coated platinum tip and a crystalline platinum substrate, where both the contact size and conductance can be inferred from the positions of atoms in the interface. We develop a method to approximate conductance based on the distance between atoms in platinum channels across the contact. Then, the contact area calculated from conductance using ballistic transport and tunneling theories is compared to that obtained using the known positions of atoms in the contact. The difference is small for very thin (<0.1 nm) or very thick (>1.0 nm) oxides, where ballistic transport and tunneling theories work well; however, the difference is significant for oxides between these limits, which is expected to be the case for platinum in many practical applications.


[Attitude to eating and body weight by 11- to 16-year-old adolescents].
Based on the data of 966 students grade five to eight, a new questionnaire--the "Eating Behaviour and Weight Problems Inventory for Children (EWI-C)"--was developed. Its 60 items are assigned to 10 subscales measuring: (1) hunger level and susceptibility to food cues, (2) importance and impact of eating on sense of well-being, (3) eating as a means of coping with emotional stress, (4) concerns about eating and weight, (5) dietary restraint, (6) attitude toward healthful nutrition, (7) attitude toward the obese, (8) pressures to eat from parents, (9) fear of weight gain, (10) figure dissatisfaction. Neither children's age nor father's educational level were related to their subscale scores to a significant degree. However, on all scales (except scale 3) important gender differences could be observed. From the youngest age groups on, mean values of girls clearly exceeded those of boys on scales 4, 5, 9 and 10, while on scales 1, 2, 7 and 8 mean scores of girls were significantly lower. To analyse relationships between EWI scores and children's relative weight, 620 students' data were combined with those of 445 adolescents attending weight-reduction programs in two obesity clinics. Boys' and girls' individual deviations from mean weight per height were used to form six groups ranging from underweight to severe obesity. In both sexes mean scores on scales 3-5, 9 and 10 significantly increased with increasing (over) weight, while a (partial) decrease could be observed on scales 2 and 8. Scores of subscales 1 and 7 proved to be independent of children's relative weight. Based on the data of the combined sample (n = 1065), norm tables with percentile ranks are provided which allow for children's sex and weight. By means of these tables the position of a boy's or girl's subscale score can be evaluated in relation to his/her weight category (underweight, normal weight, obesity, severe obesity). A child's responses on the EWI-C can be analysed by a DOS-program which is available on diskette.


Wearable Health Technology and Electronic Health Record Integration: Scoping Review and Future Directions.
Due to the adoption of electronic health records (EHRs) and legislation on meaningful use in recent decades, health systems are increasingly interdependent on EHR capabilities, offerings, and innovations to better capture patient data. A novel capability offered by health systems encompasses the integration between EHRs and wearable health technology. Although wearables have the potential to transform patient care, issues such as concerns with patient privacy, system interoperability, and patient data overload pose a challenge to the adoption of wearables by providers. This study aimed to review the landscape of wearable health technology and data integration to provider EHRs, specifically Epic, because of its prevalence among health systems. The objectives of the study were to (1) identify the current innovations and new directions in the field across start-ups, health systems, and insurance companies and (2) understand the associated challenges to inform future wearable health technology projects at other health organizations. We used a scoping process to survey existing efforts through Epic's Web-based hub and discussion forum, UserWeb, and on the general Web, PubMed, and Google Scholar. We contacted Epic, because of their position as the largest commercial EHR system, for information on published client work in the integration of patient-collected data. Results from our searches had to meet criteria such as publication date and matching relevant search terms. Numerous health institutions have started to integrate device data into patient portals. We identified the following 10 start-up organizations that have developed, or are in the process of developing, technology to enhance wearable health technology and enable EHR integration for health systems: Overlap, Royal Philips, Vivify Health, Validic, Doximity Dialer, Xealth, Redox, Conversa, Human API, and Glooko. We reported sample start-up partnerships with a total of 16 health systems in addressing challenges of the meaningful use of device data and streamlining provider workflows. We also found 4 insurance companies that encourage the growth and uptake of wearables through health tracking and incentive programs: Oscar Health, United Healthcare, Humana, and John Hancock. The future design and development of digital technology in this space will rely on continued analysis of best practices, pain points, and potential solutions to mitigate existing challenges. Although this study does not provide a full comprehensive catalog of all wearable health technology initiatives, it is representative of trends and implications for the integration of patient data into the EHR. Our work serves as an initial foundation to provide resources on implementation and workflows around wearable health technology for organizations across the health care industry.


Post-rotavirus vaccine intussusception in identical twins: A case report.
The intussusception is one of the most frequent causes of occlusive syndrome in infants and in children. (1) The mesenteric lymphadenopathy, wich is very rare post rotavirus vaccination, can cause intussusception, (2-5) especially in genetically predisposed individuals. (6) There is an association between intussusception and some classes of genotype. (7-9) Two infants aged 3 months, vaccinated against rotavirus. After about a week, one of the 2 identical infants presented inconsolable crying, vomiting, loose stools mixed with blood, and was diagnosed with bowel obstruction with intussusception. He was operated in urgency. After a few hours, his brother presented vomiting, and was admitted to our Hospital for suspected intussusception. The controls carried out have confirmed the presence of intussusception that was treated early, before the onset of severe symptoms. The incidence of post rotavirus vaccine intussusception is very low. The determining factor hypothetically might be linked to the presence of a genotype that exposes infants to a greater risk of developing mesenteric lymphadenitis and intussusception. In our case, the diagnosis of intussusception occurred in a twin, which allowed us to recognize early symptoms which accused the brother and schedule the surgery with less urgency. Our experience may want to sensitize families and pediatricians to report cases of intussusception given a theoretical familiar association. The study of the genotype could be decisive for or not to exclude the presence of a risk of invagination, thus avoiding vaccination.


Do high childcare costs and low access to Head Start and childcare subsidies limit mothers' employment? A state-level analysis.
Access to affordable childcare is crucial to mothers' employment. Yet, childcare costs and access to Head Start, childcare subsidies, and state-funded preschool vary dramatically across U.S. states. Using data from the 2016 American Community Survey five-year estimates, we apply hierarchical logistic regression models to show mothers are more likely to work in states with inexpensive childcare, higher Head Start enrollment and childcare subsidy participation, and increased availability of state-funded preschool. Childcare subsidy access is associated with higher maternal employment amongst those with lower levels of educational attainment, whereas state-funded preschool is associated with higher employment primarily among the college educated. Additionally, our analysis revealed that Head Start has a stronger association with maternal employment in states where childcare costs are high, reducing the negative relationship of employment with expensive childcare. As national discussions continue to center on the importance of childcare, our research adds evidence that public programs support maternal employment through reducing out-of-pocket childcare costs.


Transcriptomic and proteomic analyses reveal new insights into the regulation of immune pathways during adenovirus type 2 infection.
Human adenovirus (Ad) infection leads to the changes of host cell gene expression and biosynthetic processes. Transcriptomics in adenovirus type 2 (Ad2)-infected lung fibroblasts (IMR-90) cells has previously been studied using RNA sequencing. However, this study included only two time points (12 and 24 hpi) using constrained 76 bp long sequencing reads. Therefore, a more detailed study of transcription at different phases of infection using an up-graded sequencing technique is recalled. Furthermore, the correlation between transcription and protein expression needs to be addressed. In total, 3556 unique cellular genes were identified as differentially expressed at the transcriptional level with more than 2-fold changes in Ad2-infected cells as compared to non-infected cells by using paired-end sequencing. Based on the kinetics of the gene expression changes at different times after infection, these RNAs fell into 20 clusters. Among them, cellular genes involved in immune response were highly up-regulated in the early phase before becoming down-regulated in the late phase. Comparison of differentially expressed genes at transcriptional and posttranscriptional levels revealed low correlation. Particularly genes involved in cellular immune pathways showed a negative correlation. Here, we highlight the genes which expose inconsistent expression profiles with an emphasis on key factors in cellular immune pathways including NFκB, JAK/STAT, caspases and MAVS. Different from their transcriptional profiles with up- and down-regulation in the early and late phase, respectively, these proteins were up-regulated in the early phase and were sustained in the late phase. A surprising finding was that the target genes of the sustained activators failed to show response. There were features common to genes which play important roles in cellular immune pathways. Their expression was stimulated at both RNA and protein levels during the early phase. In the late phase however, their transcription was suppressed while protein levels remained stable. These results indicate that Ad2 and the host cell use different strategies to regulate cellular immune pathways. A control mechanism at the post-translational level must thus exist which is under the control of Ad2.


Information content-based gene ontology semantic similarity approaches: toward a unified framework theory.
Several approaches have been proposed for computing term information content (IC) and semantic similarity scores within the gene ontology (GO) directed acyclic graph (DAG). These approaches contributed to improving protein analyses at the functional level. Considering the recent proliferation of these approaches, a unified theory in a well-defined mathematical framework is necessary in order to provide a theoretical basis for validating these approaches. We review the existing IC-based ontological similarity approaches developed in the context of biomedical and bioinformatics fields to propose a general framework and unified description of all these measures. We have conducted an experimental evaluation to assess the impact of IC approaches, different normalization models, and correction factors on the performance of a functional similarity metric. Results reveal that considering only parents or only children of terms when assessing information content or semantic similarity scores negatively impacts the approach under consideration. This study produces a unified framework for current and future GO semantic similarity measures and provides theoretical basics for comparing different approaches. The experimental evaluation of different approaches based on different term information content models paves the way towards a solution to the issue of scoring a term's specificity in the GO DAG.


[Scientific basis for impression taking with silicone elastomers].
Starting from the chemical and physical processes which occur during the cross-linking of silicones, the author represents the material and processing properties of these impression compounds. He reports above all of the results of utilizationoriented studies which evidence the effects of different conditions of processing on the behaviour during impression taking and on the impression itself. Conclusions are drawn as to the appropriate use of silicone impression materials.


Ethylene oxide sterilization of medical devices: a review.
Ethylene oxide (EO) is a well-known sterilizing agent. However, only recently has its use significantly emerged, based on its range of applications in the field of new medical device development and sterilization. This paper describes the progress in terms of EO sterilization and concludes that it remains a promising field to explore and develop. The EO action mechanism and toxicity are analyzed, and a critical analysis is made on how it is possible to use EO sterilization for medical devices advantageously, with emphasis on cycle design and validation. One huge challenge is related with the development of mathematical models to integrate lethality to allow a continuous increase of process flexibility, without compromising its safety. The scientific community should also focus on other important issues, such as EO diffusion in different substrates, taking into account different environmental conditions both for sterilization and aeration.


The use of healthcare services for mental health problems by middle-aged and older adults.
Although mental disorders occur commonly in later life, it has been reported that older adults are reluctant to seek help for their mental health problems. The purpose of this research study was to analyze the contact with healthcare professionals, self-perceived mental health problems and unmet needs, as reported by a nationally representative sample of community-dwelling adults. We report a cross-sectional analysis of all the respondents of the Australian National Survey of Mental Health and Wellbeing aged 55 years and older (N=3178). Results indicated that 306 (9.6%) participants had a DSM-IV classifiable mental disorder based on self-identified symptoms over the preceding 12 months. Of these, 146 (48%) reported that they had not consulted a healthcare professional to deal with their mental health problems. Among those who consulted with a healthcare professional, the general practitioner was the main point of contact. Medication and psychotherapy/counseling were the most frequent form of help obtained. Informational and instrumental help, such as help to sort out practical problems and to look after oneself, were the most reported unmet needs. These results suggest a gap in the provision of healthcare services for mental health problems directed toward the specific needs of aging adults. The reported unmet needs might be met by increasing awareness amongst healthcare professionals regarding mental health problems in later stages of life and by improving the access of older people to the services commonly provided by multidisciplinary teams.


Switchable metal-to-half-metal transition at the semi-hydrogenated graphene/ferroelectric interface.
Tuning the half-metallicity of low-dimensional materials using an electric field is particularly appealing for spintronic applications but typically requires an ultra-high field, hampering practical applications. Interface engineering has been suggested as an alternative practical means to overcome this limitation and control the metal-to-half-metal transition. Here, we show from first-principles calculations that the polarization switching at the interface of semi-hydrogenated graphene (i.e., graphone) and a ferroelectric PbTiO3 layer can reversibly tune a metal to half-metal transition in graphone. Using a simple Hubbard model, this is rationalized using interface atomic orbital hybridization, which also reveals the origin of the high-quality screening of metallic graphone, preserving bulk-like stable ferroelectric polarization in the PbTiO3 film down to a thickness of two unit cells. These findings do not only open a new perspective on engineering half-metallicity at the interface of two-dimensional materials and ferroelectrics, but also identify graphone as a powerful atomically thin electrode, which holds great promise for the design of ultrafast and high integration density information-storage devices.


Overexpressed Fatty Acid Synthase in Gastrointestinal Stromal Tumors: Targeting a Progression-Associated Metabolic Driver Enhances the Antitumor Effect of Imatinib.
Purpose: In gastrointestinal stromal tumors (GIST), lipid-metabolizing enzymes remain underexplored, including fatty acid synthase (FASN).Experimental Design: Forty GISTs were quantitated for FASN mRNA abundance. FASN immunoexpression was informative in 350 GISTs, including 213 with known KIT/PDGFRA/BRAF genotypes. In imatinib-resistant FASN-overexpressing GIST cells, the roles of overexpressed FASN and FASN-targeting C75 in tumor phenotypes, apoptosis and autophagy, KIT transcription, PI3K/AKT/mTOR activation, and imatinib resistance were analyzed by RNAi or myristoylated-AKT transfection. The therapeutic relevance of dual blockade of FASN and KIT was evaluated in vivoResults:FASN mRNA abundance significantly increased from very low/low-risk to high-risk levels of NCCN guidelines (P < 0.0001). FASN overexpression was associated with a nongastric location (P = 0.05), unfavorable genotype (P = 0.005), and increased risk level (P < 0.001) and independently predicted shorter disease-free survival (P < 0.001). In vitro, FASN knockdown inhibited cell growth and migration, inactivated the PI3K/AKT/mTOR pathway, and resensitized resistant GIST cells to imatinib. C75 transcriptionally repressed the KIT promoter, downregulated KIT expression and phosphorylation, induced LC3-II and myristoylated AKT-suppressible activity of caspases 3 and 7, attenuated the PI3K/AKT/mTOR/RPS6/4E-BP1 pathway activation, and exhibited dose-dependent therapeutic additivism with imatinib. Compared with both monotherapies, the C75/imatinib combination more effectively suppressed the growth of xenografts, exhibiting decreased KIT phosphorylation, Ki-67, and phosphorylated PI3K/AKT/mTOR levels and increased TUNEL labeling.Conclusions: We have characterized the prognostic, biological, and therapeutic implications of overexpressed FASN in GISTs. C75 represses KIT transactivation, abrogates PI3K/AKT/mTOR activation, and provides a rationale for dual blockade of KIT and FASN in treating imatinib-resistant GISTs. Clin Cancer Res; 23(16); 4908-18. ©2017 AACR.


Expression changes in human skeletal muscle miRNAs following 10 days of bed rest in young healthy males.
Studies in humans show global changes in mRNA and protein expression occur in human skeletal muscle during bed rest. As microRNAs are important regulators of expression, we analysed the global microRNA expression changes in human muscle following 10 days of sustained bed rest, with the rationale that miRNAs play key roles in atrophy of skeletal muscle. We analysed expression of miRNA and selected target proteins before and after 10 days of bed rest in biopsies obtained from the vastus lateralis muscle of 6 healthy males. Fifteen of 152 miRNAs detected in human muscle tissue were differentially expressed, and all of them with exception of two were downregulated. The downregulated miRNAs include the following: miR-206, a myomir involved in function and maintenance of skeletal muscle; miR-23a, involved in insulin response and atrophy defence; and several members of the let-7 family involved in cell cycle, cell differentiation and glucose homeostasis. Predicted gene targets of these miRNAs are members of the MAPK, TNF receptor, ALK1, TGF-beta receptor and SMAD signalling pathways. All of these pathways were previously indicated to be involved in skeletal muscle response to physical inactivity. We also measured protein expression of selected miRNA targets and observed a decrease in HDAC4. Our data demonstrate that miRNAs in postural muscles are affected by sustained inactivity and unloading, as induced by prolonged bed rest, and hence are potentially involved in regulation of skeletal muscle adjustments to inactivity. We also propose new miRNAs involved in regulation of biological processes in adult human muscle.


Immunogenicity screening in protein drug development.
Most therapeutic proteins in clinical trials or on the market are, to a variable extent, immunogenic. Formation of antidrug antibodies poses a risk that should be assessed during drug development, as it possibly compromises drug safety and alters pharmacokinetics. The generation of these antibodies is critically dependent on the presence of T helper cell epitopes, which have classically been identified by in vitro methods using blood cells from human donors. As a novel development, in silico methods that identify T cell epitopes have been coming on line. These methods are relatively inexpensive and allow the mapping of epitopes from virtually all human leukocyte antigen molecules derived from a wide genetic background. In vitro assays remain important, but guided by in silico data they can focus on selected peptides and human leukocyte antigen haplotypes, thereby significantly reducing time and cost.


Ultrastructural demonstration of terminal deoxynucleotidyl transferase (TdT).
Using an electronmicroscopic peroxidase-antiperoxidase technique in combination with a three-step fixation procedure according to Willingham, the ultrastructural site of terminal deoxynucleotidyl transferase (TdT) could be shown in cells of a human lymphoblastic leukemia cell line (MOLT 4). TdT appeared in the cytoplasm of the cells associated with large membrane bound granules from 150 to 600 nm in diameter usually arranged in aggregates. TdT-positive spaces were completely segregated form the rest of the cytoplasm and seemed to be interconnected by a tubular system. No TdT was detected in the nuclei.


Effect of acute noxious stimulation to the leg or back on muscle synergies during walking.
This study aimed to examine how acute muscle pain affects muscle coordination during gait with consideration of muscle synergies (i.e., group of muscles activated in synchrony), amplitude of muscle activity and kinematics. A secondary aim was to determine whether any adaptation was specific to pain location. Sixteen participants walked on a treadmill during 5 conditions [control, low back pain (LBP), washout LBP, calf pain (CalfP), and washout CalfP]. Five muscle synergies were identified for all of the conditions. Cross-validation analysis showed that muscle synergy vectors extracted for the control condition accounted for >81% of variance accounted for from the other conditions. Muscle synergies were altered very little in some participants (n = 7 for LBP; n = 10 for CalfP), but were more affected in the others (n = 9 for LBP; n = 6 for CalfP). No systematic differences between pain locations were observed. Considering all participants, synergies related to propulsion and weight acceptance were largely unaffected by pain, whereas synergies related to other functions (trunk control and leg deceleration) were more affected. Gastrocnemii activity was less during both CalfP and LBP than control. Soleus activity was further reduced during CalfP, and this was associated with reduced plantar flexion. Some lower leg muscles exhibited adaptations depending on pain location (e.g., greater vastus lateralis and rectus femoris activity during CalfP than LBP). Overall, these changes in muscle coordination involve a participant-specific strategy that is important to further explore, as it may explain why some people are more likely to develop persistence of a painful condition.


MED12-related XLID disorders are dose-dependent of immediate early genes (IEGs) expression.
Mediator occupies a key role in protein coding genes expression in mediating the contacts between gene specific factors and the basal transcription machinery but little is known regarding the role of each Mediator subunits. Mutations in MED12 are linked with a broad spectrum of genetic disorders with X-linked intellectual disability that are difficult to range as Lujan, Opitz-Kaveggia or Ohdo syndromes. Here, we investigated several MED12 patients mutations (p.R206Q, p.N898D, p.R961W, p.N1007S, p.R1148H, p.S1165P and p.R1295H) and show that each MED12 mutations cause specific expression patterns of JUN, FOS and EGR1 immediate early genes (IEGs), reflected by the presence or absence of MED12 containing complex at their respective promoters. Moreover, the effect of MED12 mutations has cell-type specificity on IEG expression. As a consequence, the expression of late responsive genes such as the matrix metalloproteinase-3 and the RE1 silencing transcription factor implicated respectively in neural plasticity and the specific expression of neuronal genes is disturbed as documented for MED12/p.R1295H mutation. In such case, JUN and FOS failed to be properly recruited at their AP1-binding site. Our results suggest that the differences between MED12-related phenotypes are essentially the result of distinct IEGs expression patterns, the later ones depending on the accurate formation of the transcription initiation complex. This might challenge clinicians to rethink the traditional syndromes boundaries and to include genetic criterion in patients' diagnostic.


Leveraging Spatial Information in Radiology Reports for Ischemic Stroke Phenotyping.
Classifying fine-grained ischemic stroke phenotypes relies on identifying important clinical information. Radiology reports provide relevant information with context to determine such phenotype information. We focus on stroke phenotypes with location-specific information: brain region affected, laterality, stroke stage, and lacunarity. We use an existing fine-grained spatial information extraction system-Rad-SpatialNet-to identify clinically important information and apply simple domain rules on the extracted information to classify phenotypes. The performance of our proposed approach is promising (recall of 89.62% for classifying brain region and 74.11% for classifying brain region, side, and stroke stage together). Our work demonstrates that an information extraction system based on a fine-grained schema can be utilized to determine complex phenotypes with the inclusion of simple domain rules. These phenotypes have the potential to facilitate stroke research focusing on post-stroke outcome and treatment planning based on the stroke location.


Honokiol, an active compound of Magnolia officinalis, is involved in restoring normal baroreflex sensitivity in hypercholesterolemic rabbits.
This study investigated the effects of methanol extract Magnolia officinalis (MEMO) on baroreceptor reflex sensitivity (BRS) in the hypercholesterolemic rabbits and the involved molecular mechanisms. Male New Zealand white rabbits were randomly divided into Control (normal diet), Cholesterol (0.5% w/w cholesterol diet), and Magnolia groups (0.5% w/w cholesterol diet plus 1% w/w MEMO). The animals were treated with the designated diet for 4 or 8 weeks. BRS in the control of heart rate was assessed by linear regression method. After 8 weeks of treatments, plasma total cholesterol (TC) was significantly elevated in the Cholesterol/Magnolia groups. The arterial blood pressure (aBP) was increased in the Cholesterol and Magnolia groups. The depression of BRS observed in the Cholesterol group was significantly ameliorated in the Magnolia group. After L-NAME (Nω-nitro-Larginine methyl ester, 20 mg/kg, iv), the BRS of the Cholesterol group was significantly improved. Results from our in vitro study further indicated that honokiol, the principle component of MEMO, would protect human umbilical vein endothelial cells (HUVECs) from H2O2-induced damages and inhibit H2O2-induced vascular smooth muscles cells (VSMCs) proliferation, which was evident by the decreased expression of pFAK, and p-Erk1/2. The results of the present study suggested that the improvement of BRS by MEMO in the hypercholesterolemic rabbits might be mediated by the antioxidant property of MEMO as indicated by the results from the L-NAME and in vitro honokiol studies.


Field trial of active remote sensing using a high-power short-wave infrared supercontinuum laser.
Field trial results of a 5 W all-fiber broadband supercontinuum (SC) laser covering the short-wave infrared (SWIR) wavelength bands from ~1.55 to 2.35 μm are presented. The SC laser is kept on a 12 story tower at the Wright Patterson Air Force Base and propagated through the atmosphere to a target 1.6 km away. Beam quality of the SC laser after propagating through 1.6 km is studied using a SWIR camera and show a near diffraction limited beam with an M(2) value of <1.3. The SC laser is used as the illumination source to perform spectral reflectance measurements of various samples at 1.6 km, and the results are seen to be in good agreement with in-lab measurements using a conventional lamp source. Spectral stability measurements are performed after atmospheric propagation through 1.6 km and show a relative variability of ~4%-8% across the spectrum depending on the atmospheric turbulence effects. Spectral stability measurements are also performed in-lab and show a relative variability of <0.6% across the spectrum.


Resistance of ALR/Lt islets to free radical-mediated diabetogenic stress is inherited as a dominant trait.
ALS/Lt and ALR/Lt are inbred mouse strains selected for susceptibility and resistance to alloxan (AL)-induced diabetes. Within 24-h after AL administration in vivo, ALS/Lt islets were distinguished from ALR/Lt islets by more extensive necrotic changes. Within 7 days post-AL, ALS/Lt mice exhibited hyperglycemia and hypoinsulinemia, whereas ALR/Lt mice maintained normal plasma insulin and glucose levels. We have recently shown that resistance in ALR/Lt correlated with constitutively elevated systemic (and pancreatic) free radical defense status. In the present report, we examined whether ability to detoxify free radical stress extended to the level of ALR/Lt pancreatic islets. Cultured ALS/Lt islets exposed for 5 min to increasing (0-3 mmol/l) AL concentrations in vitro exhibited an 80% decline in numbers of intact islets after a subsequent 6-day culture period, as well as a 75% reduction in islet insulin content and a 94% decrease in glucose-stimulated insulin secretory capacity. In contrast, ALR/Lt islets remained viable and retained glucose-stimulated insulin secretory capacity as well as normal insulin content. This ALR/Lt islet resistance extended to hydrogen peroxide, a free radical generator whose entry into beta-cells is not dependent on glucose transporters. The elevated antioxidant defenses previously found in ALR/Lt pancreas were extended to isolated islets, which exhibited significantly higher glutathione and Cu-Zn superoxide dismutase 1 levels compared with ALS/Lt islets. A dominant genetic trait from ALR/Lt controlling this unusual AL resistance was indicated by the finding that reciprocal F1 mice of both sexes were resistant to AL administration in vivo. A backcross to ALS/Lt showed 1:1 segregation for susceptibility/resistance, indicative of a single gene controlling the phenotype. In conclusion, the ALR/Lt mouse may provide important insight into genetic mechanisms capable of rendering islets strongly resistant to free radical-mediated damage.


Ulcerative Colitis in Adults: A Review.
Ulcerative colitis (UC) is a chronic inflammatory condition of the colon, with a prevalence exceeding 400 per 100 000 in North America. Individuals with UC have a lower life expectancy and are at increased risk for colectomy and colorectal cancer. UC impairs quality of life secondary to inflammation of the colon causing chronic diarrhea and rectal bleeding. Extraintestinal manifestations, such as primary sclerosing cholangitis, occur in approximately 27% of patients with UC. People with UC require monitoring of symptoms and biomarkers of inflammation (eg, fecal calprotectin), and require colonoscopy at 8 years from diagnosis for surveillance of dysplasia. Risk stratification by disease location (eg, Montreal Classification) and disease activity (eg, Mayo Score) can guide management of UC. First-line therapy for induction and maintenance of remission of mild to moderate UC is 5-aminosalicylic acid. Moderate to severe UC may require oral corticosteroids for induction of remission as a bridge to medications that sustain remission (biologic monoclonal antibodies against tumor necrosis factor [eg, infliximab], α4β7 integrins [vedolizumab], and interleukin [IL] 12 and IL-23 [ustekinumab]) and oral small molecules that inhibit janus kinase (eg, tofacitinib) or modulate sphingosine-1-phosphate (ozanimod). Despite advances in medical therapies, the highest response to these treatments ranges from 30% to 60% in clinical trials. Within 5 years of diagnosis, approximately 20% of patients with UC are hospitalized and approximately 7% undergo colectomy. The risk of colorectal cancer after 20 years of disease duration is 4.5%, and people with UC have a 1.7-fold higher risk for colorectal cancer compared with the general population. Life expectancy in people with UC is approximately 80.5 years for females and 76.7 years for males, which is approximately 5 years shorter than people without UC. UC affects approximately 400 of every 100 000 people in North America. An effective treatment for mild to moderate UC is 5-aminosalicylic acid, whereas moderate to severe UC can be treated with advanced therapies that target specific inflammation pathways, including monoclonal antibodies to tumor necrosis factor, α4β7 integrins, and IL-12 and IL-23 cytokines, as well as oral small molecule therapies targeting janus kinase or sphingosine-1-phosphate.


[The dependence of the adaptive effect of brief stress exposures on the thyroid state of the body].
The effectiveness of adaptation to short-term stresses with regard to the level of thyroid hormones was investigated in experiments on isolated hearts of 94 female rats. Adaptation to stress in euthyroid rats (daily immobilization for 15 days) prevented a 2-fold decrease in the blood concentrations of T3 and thyroxine, caused by 6-hour immobilization stress and determined by radioimmunoassay. In hypothyroid rats multiple sessions of short-term immobilization in itself caused a decrease in the coronary dilating reserve by 29%, in the maximum coronary blood flow developing after 60-second discontinuation of perfusion, by 31%, an increase in intraventricular diastolic pressure; subsequent 6-hour immobilization resulted in a further drop of the coronary reserve, maximum coronary blood flow and an increase in diastolic pressure. Thus the effect of preventing stress changes of the coronary blood flow and myocardial contractility by adaptation to short-term stresses depends on body thyroid function.


Sensitivity of Providence to antiseptics and disinfectants.
Seventy isolates of Providence were obtained from urinary tract infections in patients from five hospitals. They were all identified as Providence B (Providencia stuartii). A typing system was devised for these organisms based on their production of and sensitivity to bacteriophage and bacteriocins. Using this system, 27 distinct strains were recognized, and it was clear that many patients had been infected with the same strain. The minimum inhibitory concentrations of six antiseptics and disinfectants for these strains were determined. Many of the strains were up to 300-fold more resistant to the cationic antiseptics chlorhexidine, cetrimide, and Resiguard than the control strain Escherichia coli NCTC 10418. The sensitivities of the Providence strains to Hycolin, glutaraldehyde, and phenyl mercuric nitrate, however, were similar to those of the control organism. There were significant correlations between the sensitivities of the strains to the three cationic antiseptics. The loss of chlorhexidine resistance from a Providence strain after exposure of the cells to N-methyl-N' nitro N nitrosoguanidine resulted in a concomitant loss of resistance to cetrimide and Resiguard, but there was no significant increase in the sensitivity of the mutants to the other three antibacterials.


Mapping of dominant B-cell epitopes of a human zona pellucida protein (ZP1).
Zona pellucida (ZP) glycoproteins contain numerous antigenic determinants including carbohydrate, protein, and conformational epitopes; and the immunogenicity of these complex glycoproteins varies in different mammalian hosts. Studies have now shown that antibodies from primates immunized with a cDNA-expressed recombinant rabbit ZP protein (the homologue of the human ZP1 [hZP1]) inhibit sperm binding to the ZP without altering ovarian function, unlike immunization with ZP3 and ZP2 protein families. The ZP1 protein or peptides derived from it (recombinant or synthetic) are therefore primary candidates for use in designing safe and reversible human and animal contraceptive vaccines. In order to define peptide epitope(s) that may be critical for eliciting an immune response sufficient to effect immunological contraception without causing any adverse effects on ovarian physiology, studies have been carried out to identify immunodominant B-cell epitopes of the ZP1 protein. The amino acid sequence of the hZP1 was used to design a set of 94 (15-mer) biotinylated peptides having an overlap of 9 amino acids. Using these peptides in a modified enzyme-linked immunoassay, antibodies in sera from rabbits or baboons immunized with native porcine ZP protein were screened for ZP1 peptide recognition. These studies demonstrate that there are a limited number of peptides recognized by primate antibodies but that the overlapping peptides sharing the sequence GPLTLELQI are recognized by both rabbit and baboon antibodies regardless of the adjuvant system used to induce the immune response. This peptide is 100% conserved in amino acid sequence between the human and pig, although the rabbit protein has two conserved amino acid substitutions (100% similar, 77% identical). Because this peptide is immunogenic as well as antigenic in primates, it could play a major role in the development of human contraceptive vaccines.


Synchronization through nonreciprocal connections in a hybrid hippocampus microcircuit.
Synchronization among neurons is thought to arise from the interplay between excitation and inhibition; however, the connectivity rules that contribute to synchronization are still unknown. We studied these issues in hippocampal CA1 microcircuits using paired patch clamp recordings and real time computing. By virtually connecting a model interneuron with two pyramidal cells (PCs), we were able to test the importance of connectivity in synchronizing pyramidal cell activity. Our results show that a circuit with a nonreciprocal connection between pyramidal cells and no feedback from PCs to the virtual interneuron produced the greatest level of synchronization and mutual information between PC spiking activity. Moreover, we investigated the role of intrinsic membrane properties contributing to synchronization where the application of a specific ion channel blocker, ZD7288 dramatically impaired PC synchronization. Additionally, background synaptic activity, in particular arising from NMDA receptors, has a large impact on the synchrony observed in the aforementioned circuit. Our results give new insights to the basic connection paradigms of microcircuits that lead to coordination and the formation of assemblies.


Specific targeting of the EBV lytic phase protein BNLF2a to the transporter associated with antigen processing results in impairment of HLA class I-restricted antigen presentation.
EBV persists for life in the human host while facing vigorous antiviral responses that are induced upon primary infection. This persistence supports the idea that herpesviruses have acquired dedicated functions to avoid immune elimination. The recently identified EBV gene product BNLF2a blocks TAP. As a result, reduced amounts of peptides are transported by TAP from the cytoplasm into the endoplasmic reticulum (ER) lumen for binding to newly synthesized HLA class I molecules. Thus, BNLF2a perturbs detection by cytotoxic T cells. The 60-aa-long BNLF2a protein prevents the binding of both peptides and ATP to TAP, yet further mechanistic insight is, to date, lacking. In this study, we report that EBV BNLF2a represents a membrane-associated protein that colocalizes with its target TAP in subcellular compartments, primarily the ER. In cells devoid of TAP, expression levels of BNLF2a protein are greatly diminished, while ER localization of the remaining BNLF2a is retained. For interactions of BNLF2a with the HLA class I peptide-loading complex, the presence of TAP2 is essential, whereas tapasin is dispensible. Importantly, we now show that in B cells supporting EBV lytic replication, the BNLF2a protein is expressed early in infection, colocalizing and associating with the peptide-loading complex. These results imply that, during productive EBV infection, BNLF2a contributes to TAP inhibition and surface HLA class I down-regulation. In this way, EBV BNLF2a-mediated evasion from HLA class I-restricted T cell immunity contributes to creating a window for undetected virus production.


Comparison of bilateral internal thoracic artery revascularization using in situ or Y graft configurations: a prospective randomized clinical, functional, and angiographic midterm evaluation.
Bilateral internal thoracic arteries (BITA) demonstrated superiority over other grafts to the left coronary system in terms of patency and survival benefit. Several BITA configurations are proposed for left-sided myocardial revascularization, but the ideal BITA assemblage is still unidentified. From 03/2003 to 08/2006, 1297 consecutive patients underwent isolated bypass surgery in our institution. 481 patients met the inclusion criteria for randomization, and 304 (64%) were randomized. Patients were allocated to BITA in situ grafting (n=147) or Y configuration (n=152) then evaluated for clinical, functional, and angiographic outcome after 6 months and 3 years. Patient telephone interviews were conducted every 3 months and a stress test performed twice yearly under the referring cardiologist's supervision. Angiographic follow-up was performed 6 months after surgery. The primary and secondary end points were, respectively, major adverse cerebrocardiovascular events (MACCE) and the proportion of ITA grafts that were completely occluded at follow-up angiography. More arterial anastomoses were performed in patients randomized to the Y than the in situ configuration (3.2 versus 2.4; P<0.001). No significant difference between the 2 groups in terms of hospital mortality or morbidity was found. At follow-up, there was no significant difference in any MACCE rate between the 2 groups. 450 out of 464 anastomosis (97%) in the BITA Y group and 287 of 295 (97%) in the BITA in situ group were controlled patent (P=0.99). Excellent patency rates were achieved using both BITA configurations with no significant differences in terms of MACCE up to 19 months postoperatively, but longer-term results remain to be established.


Warmth from skin-to-skin contact with mother is essential for the acquisition of filial huddling preference in preweanling rats.
During a single, 2-hr session with a scented foster dam, preweanling rat pups form an affiliative attraction to an odor associated with the maternal caregiver, manifest as a huddling preference. To identify maternal stimuli that induce this filial preference, we quantitatively examined behavioral interactions during odor conditioning. Bout duration of skin-to-skin (STS) contact was positively associated with the preference. In contrast, simple physical contact and anogenital licking were not significantly related to the preference. The frequency of nonanogenital licking was negatively associated with the preference as well as with bout duration of STS contact. When odor conditioning was conducted with a warm cylinder, ambient warmth, or stroking as the unconditioned stimulus, only pups exposed to the warm cylinder exhibited a preference for the conditioned odor. These results suggest a positive, affiliative effect of maternal STS contact on pup filial preference, which may be disrupted by maternal licking.


Hippocampal shape analysis using medial surfaces.
In magnetic resonance imaging (MRI) research, significant attention has been paid to the analysis of the hippocampus (HC) within the medial temporal lobe because of its importance in memory and learning, and its role in neurodegenerative diseases. Manual segmentation protocols have established a volume decline in the HC in conjunction with Alzheimer's disease, epilepsy, post-traumatic stress disorder, and depression. Furthermore, recent studies have investigated age-related changes of HC volume which show an interaction with gender; in early adulthood, volume reduction of the HC is found in men but not in women. In this paper, we investigated gender differences in normal subjects in young adulthood by employing a shape analysis of the HC using medial surfaces. For each subject, the most prominent medial manifold of the HC was extracted and flattened. The flattened sheets were then registered using both a rigid and a non-rigid alignment technique, and the medial surface radius was expressed as a height function over them. This allowed for an investigation of the association between subject variables and the local width of the HC. With regard to the effects of age and gender, it could be shown that the previously observed gender differences were mostly due to volume loss in males in the lateral areas of the HC head and tail. We suggest that the analysis of HC shape using medial surfaces might thus serve as a complimentary technique to investigate group differences to the established segmentation protocols for volume quantification in MRI.


Arrival and diversification of caviomorph rodents and platyrrhine primates in South America.
Platyrrhine primates and caviomorph rodents are clades of mammals that colonized South America during its period of isolation from the other continents, between 100 and 3 million years ago (Mya). Until now, no molecular study investigated the timing of the South American colonization by these two lineages with the same molecular data set. Using sequences from three nuclear genes (ADRA2B, vWF, and IRBP, both separate and combined) from 60 species, and eight fossil calibration constraints, we estimated the times of origin and diversification of platyrrhines and caviomorphs via a Bayesian relaxed molecular clock approach. To account for the possible effect of an accelerated rate of evolution of the IRBP gene along the branch leading to the anthropoids, we performed the datings with and without IRBP (3768 sites and 2469 sites, respectively). The time window for the colonization of South America by primates and by rodents is demarcated by the dates of origin (upper bound) and radiation (lower bound) of platyrrhines and caviomorphs. According to this approach, platyrrhine primates colonized South America between 37.0 +/- 3.0 Mya (or 38.9 +/- 4.0 Mya without IRBP) and 16.8 +/- 2.3 (or 20.1 +/- 3.3) Mya, and caviomorph rodents between 45.4 +/- 4.1 (or 43.7 +/- 4.8) Mya and 36.7 +/- 3.7 (or 35.8 +/- 4.3) Mya. Considering both the fossil record and these molecular datings, the favored scenarios are a trans-Atlantic migration of primates from Africa at the end of the Eocene or beginning of the Oligocene, and a colonization of South America by rodents during the Middle or Late Eocene. Based on our nuclear DNA data, we cannot rule out the possibility of a concomitant arrival of primates and rodents in South America. The caviomorphs radiated soon after their arrival, before the Oligocene glaciations, and these early caviomorph lineages persisted until the present. By contrast, few platyrrhine fossils are known in the Oligocene, and the present-day taxa are the result of a quite recent, Early Miocene diversification.


Progression in smoldering Waldenstrom macroglobulinemia: long-term results.
The purpose of this study was to define the risk of progression and survival of patients with smoldering Waldenström macroglobulinemia (SWM). SWM is defined clinically as having a serum monoclonal IgM protein≥3 g/dL and/or≥10% bone marrow lymphoplasmacytic infiltration but no evidence of end-organ damage (anemia, constitutional symptoms, hyperviscosity, lymphadenopathy, or hepatosplenomegaly). We searched a computerized database and reviewed the medical records of all patients at Mayo Clinic who fulfilled the criteria of SWM between 1974 and 1995. During 285 cumulative person-years of follow-up of the 48 patients with SWM (median, 15.4 years), 34 (71%) progressed to symptomatic Waldenström macroglobulinemia (WM) requiring treatment, one to primary amyloidosis, and one to lymphoma (total, 75%). The cumulative probability of progression to symptomatic WM, amyloidosis, or lymphoma was 6% at 1 year, 39% at 3 years, 59% at 5 years, and 68% at 10 years. The major risk factors for progression were percentage of lymphoplasmacytic cells in the bone marrow, size of the serum M-spike, and the hemoglobin value. Patients with SWM should be followed and not treated until symptomatic WM develops. Treatment on a clinical trial for those at greatest risk of progression should be considered.


Harnessing Endogenous Cellular Mechanisms for Bone Repair.
Although autologous tissue transplantation represents a valid approach for bone repair, it has encountered crucial barriers in therapeutic translation, not least the invasive process necessary for stem cell isolation. In recent years, the scientific community has made significant strides for identifying new treatment options, and great emphasis has been placed on the tight interaction between skeletal and immune system in modulating the outcome of bone repair. Within the context of specific injury environmental cues, the cross talk among inflammatory cells and tissue resident and/or circulating progenitor cells is crucial to finely coordinate repair and remodeling processes. The appropriate modulation of the inflammatory response can now be considered a new trend in the field of regenerative medicine, as it raises the attracting possibility to enhance endogenous progenitor cell functions, finally leading to tissue repair. Therefore, new treatment options have been developed considering the wide spectrum of bone-inflammation interplay, considering in particular the cell intrinsic cues responsible for the modulation of the injured environment. In this review, we will provide a panoramic overview focusing on novel findings developed to uphold endogenous bone repair.


Revised Enskog theory for Mie fluids: Prediction of diffusion coefficients, thermal diffusion coefficients, viscosities, and thermal conductivities.
Since the 1920s, the Enskog solutions to the Boltzmann equation have provided a route to predicting the transport properties of dilute gas mixtures. At higher densities, predictions have been limited to gases of hard spheres. In this work, we present a revised Enskog theory for multicomponent mixtures of Mie fluids, where the Barker-Henderson perturbation theory is used to calculate the radial distribution function at contact. With parameters of the Mie-potentials regressed to equilibrium properties, the theory is fully predictive for transport properties. The presented framework offers a link between the Mie potential and transport properties at elevated densities, giving accurate predictions for real fluids. For mixtures of noble gases, diffusion coefficients from experiments are reproduced within ±4%. For hydrogen, the predicted self-diffusion coefficient is within 10% of experimental data up to 200 MPa and at temperatures above 171 K. Binary diffusion coefficients of the CO2/CH4 mixture from simulations are reproduced within 20% at pressures up to 14.7 MPa. Except for xenon in the vicinity of the critical point, the thermal conductivity of noble gases and their mixtures is reproduced within 10% of the experimental data. For other molecules than noble gases, the temperature dependence of the thermal conductivity is under-predicted, while the density dependence appears to be correctly predicted. Predictions of the viscosity are within ±10% of the experimental data for methane, nitrogen, and argon up to 300 bar, for temperatures ranging from 233 to 523 K. At pressures up to 500 bar and temperatures from 200 to 800 K, the predictions are within ±15% of the most accurate correlation for the viscosity of air. Comparing the theory to an extensive set of measurements of thermal diffusion ratios, we find that 49% of the model predictions are within ±20% of the reported measurements. The predicted thermal diffusion factor differs by less than 15% from the simulation results of Lennard-Jones mixtures, even at densities well exceeding the critical density.


Parkinson disease: the enteric nervous system spills its guts.
Lewy pathology in Parkinson disease (PD) extends well beyond the CNS, also affecting peripheral autonomic neuronal circuits, especially the enteric nervous system (ENS). The ENS is an integrative neuronal network also referred to as "the brain in the gut" because of its similarities to the CNS. We have recently shown that the ENS can be readily analyzed using routine colonic biopsies. This led us to propose that the ENS could represent a unique window to assess the neuropathology in living patients with PD. In this perspective, we discuss current evidence which indicates that the presence of ENS pathology may by exploited to improve our understanding and management of PD and likely other neurodegenerative disorders.


Evidence of microplastic ingestion in the shark Galeus melastomus Rafinesque, 1810 in the continental shelf off the western Mediterranean Sea.
Microplastic (<5 mm) ingestion has been recorded in Galeus melastomus, the blackmouth catshark, around the Balearic Islands. In total 125 individuals were analyzed for microplastic ingestion. Results have shown that 16.80% of the specimens had ingested a mean value of 0.34 ± 0.07 microplastics/individual. Stomach fullness index ranged from 0.86 to 38.89% and regression analyses showed that fuller stomachs contained more microplastics. A higher quantity of filament type microplastics were identified compared to granular or hard plastic type. No significant differences were given between ingestion values of two locations over the continental shelf providing further evidence of the ubiquitous distribution of microplastics. The findings in this study reflect the availability of this man made contaminant to marine species in seafloor habitats. Based on results from this study, data on microplastic ingestion could be used to study trends in the amount and composition of litter ingested by marine animals in accordance with descriptor 10 of the Marine Strategy Framework Directive.


Primary thyroid MALT lymphoma with a probable brain secondary in a male patient: A case report.
Primary Mucosa-associated lymphoid tissue (MALT) lymphoma of the thyroid is a rare tumor. A previously well male in his 50s presented to our institution with difficulty in breathing and sleep apnea. He was diagnosed with a large retrosternal multinodular goiter with level 2 unilateral cervical lymphadenopathy. Fine needle aspiration cytology of the thyroid revealed chronic thyroiditis and the enlarged lymph node cytology was inconclusive. He underwent total thyroidectomy and level VI bilateral cervical lymph node clearance. The histology revealed an extra-nodal marginal zone lymphoma of MALT. A whole-body CT scan did not demonstrate any other primary site. The patient received 4 cycles of local radiotherapy. Subsequently, he was diagnosed with a brain tumor not amenable to surgical interventions following persistent headaches. He died shortly after due to complications of probable cerebral metastasis. MALT lymphomas of the thyroid carry a good prognosis; however, no universal guidance exists regarding the optimal therapy and follow-up. This case report highlights the importance of early diagnosis, identification of poor prognostic factors, and patient-tailored therapy and follow-up.


[Endogenic intoxication and free radical oxidation in urolithiasis].
The aim of the study was elucidation of relations between endogenic intoxication (EI) and free radical oxidation (FRO) in patients with urolithiasis and influence of EI and FRO on development of urolithiasis. The study included 35 urolithiasis patients (the study group) and 35 healthy controls aged 30 to 72 years. EI was assessed by content of middle molecular peptides (MMP) in the blood serum and quality-quantity composition of organic substances adsorbed on the surface of erythrocytic membrane. Investigation of adsorbed organic substances on erythrocytic membrane was conducted by assessment of adsorption-transport function of erythrocytes (ATFE) defined by means of erythrocytic wash-outs (EW) obtained according to R. A. Gareev technique (1998). Data processing was made with statistic package Microsoft Excel-2000 in S. N. Lapach modification (2000). A comparative level of MMP washed from erythrocytic membranes in urolithiasis patients is significantly elevated (p < 0.002). Total content of organic substances in the blood of urolithiasis patients is significantly increased (p < 0.01) due to their erythrocytic component. The study group had enhanced FRO processes (p < 0.01). A significant direct correlation was found between LPO product and EW, MMP (r = from +0.42 to +0.61; p < 0.05), inverse correlation between lipid peroxidation (LPO) and EW, MMP (r = from -0.54 to -0.62; p < 0.05). Adirect correlation exists between MMP level and EW (r = from +0.82 to +0.92, p < 0.01). Thus, urolithiasis patients have significant enhancement of LPO and EI. LPO activity in the blood of urolithiasis patients correlates with EI.