Acute complications of pulmonary artery catheter insertion in critically ill patients.
Of 142 critically ill patients undergoing pulmonary artery catheter (PAC) insertion, 1.4% suffered pneumothorax and 7.7% experienced arterial puncture during central venous access. Catheterization was successful in all cases; however, 8.4% of patients required special maneuvers for pulmonary artery cannulation. The 52.3% incidence of cardiac arrhythmias during PAC insertion was primarily due to ventricular arrhythmia (VA), which was more common among patients with complicated myocardial infarction (p less than .01) and less common in patients with sepsis (p less than .05). The development of VA was significantly related to the duration of PAC insertion. Our study suggests that PAC placement carries certain risks and complications which should be weighed against the advantages of a PAC in each patient.


Patterns of bilingual language use and response inhibition: A test of the adaptive control hypothesis.
Given prior studies that provided inconsistent results, there is an ongoing debate on the issue of whether bilingualism benefits cognitive control. We tested the Adaptive Control Hypothesis, according to which only the intense use of different languages in the same situation without mixing them in single utterances (called dual-language context) confers a bilingual advantage in response inhibition. In a large-scale correlational study, we attempted to circumvent several pitfalls of previous research on the bilingual advantage by testing a relatively large sample of participants and employing a more reliable and valid measurement of constructs (i.e., latent variable approach accompanied by Bayesian estimation). Our results do not support the Adaptive Control Hypothesis' prediction: the intensity of the dual-language context experience was unrelated to the efficiency of response inhibition in bilinguals. The results suggest that the Adaptive Control Hypothesis is not likely to account for the inconsistent results regarding the bilingual advantage hypothesis, at least in the case of the response-inhibition mechanism. At the same time, the study points to the problem of measuring the response-inhibition construct at the behavioral level. No evidence for a robust response-inhibition construct adds to the growing skepticism on this issue in the literature.


Pyranocarbazoles from Murraya koenigii (L.) Spreng. as antimicrobial agents.
The bioassay guided fractionation of methanolic extract of Murraya koenigii (L.) Spreng. leaves resulted in the isolation of seven pyranocarbazoles. These were evaluated against four bacterial strains and ten Candida sp. including two matched pair of fluconazole sensitive/resistant clinical isolates. Out of seven, three i.e. Koenine (mk279), Koenigine (mk309) and Mahanine (mk347) exhibited significant antibacterial activity MIC90 3.12-12.5 μg/mL against bacterial strains Streptococcus aureus and Klebsiella pneumonia compared with standard drug Kanamycin MIC90 12.5 μg/mL. However, only mk309 was found active against variety of Candida species MIC90 12.5-100 μg/mL. It was observed that hydroxylation at C-6 and C-7 positions in the studied pyranocarbazoles activate the bioactivity. Simultaneously, decrease in Log P value compares with -H and -O-CH3 substituted derivatives. The study is focused on selective antifungal and antibacterial activity of pyranocarbazoles on bacterial strains S. aureus, K. pneumonia and variety of Candida species with structure activity relationship observations.


Prostatic steroid binding protein: organisation of C1 and C2 genes.
Prostatic steroid binding protein, whose expression is stimulated by androgens, consists of two subunits: one containing the polypeptides C1 and C3 and the other containing C2 and C3. We have characterised genomic clones containing the C1 and C2 genes by restriction enzyme analysis and DNA sequencing. Both genes are 3.2 Kb, have similar exon/intron arrangements and share considerable DNA sequence homologies in their coding regions, intervening sequences and 5' upstream DNA sequence which suggests that they have probably arisen from the duplication of an ancestral gene. The 5' termini of C1 and C2 mRNA have been mapped; the sequence TATAAA appears 30 nucleotides upstream but a CAAT-like sequence at -60 - -80 is absent. Finally, homologous human genes have not been detected.


Role of immune cells in protection against and control of reovirus infection in neonatal mice.
We studied the role of T cells in resistance to reovirus intestinal and central nervous system infection. Transfer of reovirus-immune adult spleen cells protected neonatal mice from (i) lethal infection with reovirus serotype 3 Dearing (T3D, footpad inoculation) and serotype 3 clone 9 (T3C9, oral inoculation) and (ii) hydrocephalus caused by serotype 1 Lang (T1L, intracranial [i.c.] inoculation). Cell-mediated protection was not serotype specific. While immune cells protected against T1L i.c., they failed to protect against 1/5,000 of the dose of T3D i.c. Two types of experiments showed that both CD4 and CD8 T cells are involved in reovirus resistance. First, immune cell-mediated protection against T3D was abrogated by in vivo treatment with anti-CD4 monoclonal antibody (MAb) and significantly inhibited by in vivo treatment with anti-CD8 MAb. Second, T3C9-infected neonatal mice treated with anti-CD4 and/or anti-CD8 developed a novel disease phenotype, an oily hair syndrome, associated with severe hepatobiliary pathology and increased viral titer in heart and liver. Immune cells and an MAb to the cell attachment protein sigma 1 (MAb G5) protected by different mechanisms. Immune cells were more effective than sigma 1 MAb G5 at controlling primary replication, while sigma 1 MAb G5 was more effective than immune cells at inhibiting neural spread of virus. We conclude that both CD4 and CD8 T cells are important for reovirus resistance, that cells and antibody act preferentially at different stages in pathogenesis in vivo, and that adoptively transferred immune cells can protect both the central nervous system and intestine.


[Adrenergic regulation of adenylate cyclase activity in the myocardium of rats of different ages].
The mechanism of regulation of adenylate cyclase activity has been studied in adult and old rats. The basal and fluoride-stimulated adenylate cyclase activity was found to remain unchanged, while the epinephrine-stimulated enzyme activity rised in myocardium of adult and old rats. In chemical denervation the basal and stimulated activity of adenylate cyclase increased irrespective of age, the basal and epinephrine-stimulated activity of the enzyme was significantly higher in old rats. The age-related differences in regulation of myocardial adenylate cyclase appear to be determined by changes in receptor-mediated properties of adenylate cyclase complex. This phenomenon is responsible for peculiarities of the humoral regulation of the heart functions during ageing.


[A new approach to treatment selection in dogs with cruciate ligament rupture: patient-specific treatment recommendations].
Cranial cruciate ligament rupture is one of the most important diseases in canine orthopedics. Despite the frequent occurrence of the disease and the extensive literature available, there is still controversy about the best treatment method. The aim of this review article is to present a new, more specific approach to treatment selection in dogs with cranial cruciate ligament rupture. Patients are divided into different groups and particular treatment methods are then recommended according to group membership. In order to develop the treatment recommendations, the patient groups were initially defined based on criteria that are important for treatment selection, such as type of cranial cruciate ligament rupture, chronicity, degree of instability, size and weight of the patient, stage of osteoarthritis, the presence of bone deformities, concurrent medial patellar luxation or rotational instability. A detailed literature search was conducted through MEDLINE/PUBMED; CAB Abstracts, Google -Scholar and in conference proceedings abstracts from 1990-2019. Based on the available literature, treatment recommendations were developed for each patient group. These patient group-specific recommendations based on best available evidence are intended to simplify the decision-making process for treatment selection in dogs with cranial cruciate ligament disease.


Characterization of cardiac troponin subunit release into serum after acute myocardial infarction and comparison of assays for troponin T and I. American Association for Clinical Chemistry Subcommittee on cTnI Standardization.
We examined the release of cardiac troponin T (cTnT) and I (cTnI) into the blood of patients after acute myocardial infarction (AMI). Three postAMI serum samples were applied in separate analytical runs onto a calibrated gel filtration column (Sephacryl S-200), and the proteins were separated by molecular weight. Using commercial cTnT and cTnI assays measured on collected fractions, we found that troponin was released into blood as a ternary complex of cTnT-I-C, a binary complex of cTnI-C, and free cTnT, with no free cTnI within the limits of the analytical methodologies. The serum samples were also examined after incubation with EDTA and heparin. EDTA broke up troponin complexes into individual subunits, whereas heparin had no effect on the assays tested. We added free cTnC subunits to 24 AMI serum samples and found no marked increase in the total cTnI concentrations, using an immunoassay that gave higher values for the cTnI-C complex than free cTnI. To characterize the cross-reactivity of cTnT and cTnI assays, purified troponin standards in nine different forms were prepared, added to serum and plasma pools, and tested in nine quantitative commercial and pre-market assays for cTnI and one approved assay for cTnT. All nine cTnI assays recognized each of the troponin I forms (complexed and free). In five of these assays, the relative responses for cTnI were nearly equimolar. For the remainder, the response was substantially greater for complexed cTnI than for free cTnI. Moreover, there was a substantial difference in the absolute concentration of results between cTnI assays. The commercial cTnT assay recognized binary and ternary complexes of troponin on a near equimolar basis. We conclude that all assays are useful for detection of cardiac injury. However, there are differences in absolute cTnI results due to a lack of mass standardization and heterogeneity in the cross-reactivities of antibodies to various troponin I forms.


Genotypes of chronic progressive external ophthalmoplegia in a large adult-onset cohort.
Chronic progressive external ophthalmoplegia (CPEO) is a common presentation of mitochondrial disease. We performed a retrospective evaluation of the molecular genetic testing and genotype-phenotype correlations in a large cohort of adult-onset CPEO patients (N = 111). One hundred percent of patients tested had at least one mitochondrial DNA (mtDNA) deletion. Genetic testing of nuclear genes encoding mitochondrial proteins identified pathogenic/likely pathogenic variants likely to be associated with CPEO in 7.6% of patients. As expected, the nuclear gene most associated with DNA variation was POLG. A single likely pathogenic mitochondrial DNA variant (m.12278T>C) was identified in two unrelated patients. No significant differences were noted in the clinical phenotypes of patients with pathogenic or likely pathogenic nuclear variants in comparison to those with negative nuclear gene testing. Analysis of deletion size and heteroplasmy in muscle-derived mtDNA showed significant correlations with age of symptom onset but not disease severity (number of canonical CPEO features). Results suggest that smaller mtDNA deletions (p = 0.0127, r2 = 0.1201) and higher heteroplasmy of single mtDNA deletions (p = 0.0112, r2 = 0.2483) are associated with an earlier age of onset in CPEO patients.


Ultrasonographic assessment of bladder neck mobility in continent bitches and bitches with urinary incontinence attributable to urethral sphincter mechanism incompetence.
To determine bladder neck positional changes between standing and recumbent positions in bitches and whether change is related to continence status or general anesthesia, or both, and to evaluate reproducibility of measurements. 45 continent animals and 46 incontinent bitches with urethral sphincter mechanism incompetence (SMI). Distance between the bladder neck and perineal skin was measured ultrasonographically via the perineum while each dog was conscious in standing and right lateral recumbency and in right lateral recumbency under general anesthesia. Measurements of the bladder neck position obtained in right lateral recumbency under anesthesia were compared with radiographic measurements of the same parameter. Reproducibility of the ultrasonographic measurements of the distance between bladder neck and perineum was assessed on 3 occasions for each position in 50 dogs. In all dogs, differences in bladder neck position between standing and recumbent conscious-associated positions were not significant. However, caudal bladder neck movement between standing conscious- and recumbent anesthesia-associated positions and between recumbent conscious- and anesthesia-associated positions was significant for all dogs. Incontinent bitches had greater degree of caudal bladder movement during anesthesia. Mean difference in bladder neck positions between recumbent conscious- and recumbent anesthesia-associated positions was 0.24 cm in continent, compared with 0.73 cm in incontinent, bitches. Radiographic measurements were significantly greater than ultrasonographic measurements. Differences between repeated measurements for standing position were not significant, but those for recumbent conscious- and recumbent anesthesia-associated positions were significant. Additional vesicourethral support mechanisms in continent dogs, are deficient in bitches with SMI, allowing the bladder neck to move further caudad.


Options for treating carbapenem-resistant Enterobacteriaceae.
To address the therapeutic management of carbapenem-resistant Enterobacteriaceae on the basis of literature of the last 12 months. Retrospective and prospective (nonrandomized noncontrolled) studies provide data regarding the management of infections due to carbapenem-resistant Enterobacteriaceae. The combination of a carbapenem with colistin or high-dose tigecycline or aminoglycoside or even triple carbapenem-containing combinations if the minimum inhibitory concentration (MIC) range of carbapenem (meropenem and imipenem) resistance is 8 mg/l or less seems to have an advantage over monotherapy with either colistin or tigecycline or fosfomycin. For Enterobacteriaceae with MIC for carbapenems over 8 mg/l, combination regimens involve colistin, tigecycline usually administered in a double dose than that suggested by its manufacturer, fosfomycin and aminoglycosides in various combinations. Suggestions based on the limited literature cannot be made safely. Combination regimens involving carbapenems for Enterobacteriaceae with MICs 8 mg/l or less for carbapenems (in dual combination with colistin or high-dose tigecycline or aminoglycoside or even triple combinations) seem to confer some therapeutic advantage over monotherapy. For Enterobacteriaceae with higher than the above-mentioned MICs, a combination of two or even three antibiotics among colistin, high-dose tigecycline, aminoglycoside and fosfomycin seems to confer decreased mortality.


The Role of Carboplatin in the Neoadjuvant Chemotherapy Treatment of Triple Negative Breast Cancer.
Triple negative breast (TNBC) cancer constitutes a heterogeneous group of disease with histologic and molecular differences. Complete pathologic response to neoadjuvant chemotherapy (NACT) in TNBC is associated with improved outcomes. Efforts have been made in identifying drug combinations that will increase the response rate to preoperative chemotherapy. In this review we present recent studies that have incorporated carboplatin (Cb) in the NACT of TNBC. We discuss the homologous recombination deficiency score and the somatic or germline mutation for BRCA as potential biomarkers for future selection of patients that could benefit from the addition of Cb to NACT.


Two-site immunofluorometric assay for pregnancy-specific beta 1-glycoprotein (SP1).
We describe an immunoassay for human pregnancy-specific beta 1-glycoprotein by use of fluorescein-labeled antibodies to this protein. The technique is generally similar to that of a "two-site" immunoradiometric assay but has the advantage of stability of tracer and avoidance of radiation hazard. However, the procedure requires relatively large quantities of antibody. Furthermore, the within-assay coefficient of variation (11.8%) is greater than that for radioimmunoassay (6.5%). This results from imprecision of the end-point detection, and some inherent difficulties in optimizing a "two-site" immunometric system. Concentrations of endogenous material as measured by the fluorometric assay are 1.9-fold higher than those obtained with radioimmunoassay, probably because the former detects multiple forms of this heterogeneous material.


Conserved regulation and role of Pitx2 in situs-specific morphogenesis of visceral organs.
Pitx2 is expressed in developing visceral organs on the left side and is implicated in left-right (LR) asymmetric organogenesis. The asymmetric expression of Pitx2 is controlled by an intronic enhancer (ASE) that contains multiple Foxh1-binding sites and an Nkx2-binding site. These binding sites are essential and sufficient for asymmetric enhancer activity and are evolutionarily conserved among vertebrates. We now show that mice that lack the ASE of Pitx2 (Pitx2(Delta)(ASE/)(Delta)(ASE) mice) fail to manifest left-sided Pitx2 expression and exhibit laterality defects in most visceral organs, although the position of the stomach and heart looping remain unaffected. Asymmetric Pitx2 expression in some domains, such as the common cardinal vein, was found to be induced by Nodal signaling but to be independent of the ASE of Pitx2. Expression of Pitx2 appears to be repressed in a large portion of the heart ventricle and atrioventricular canal of wild-type mice by a negative feedback mechanism at a time when the gene is still expressed in its other domains. Rescue of the early phase of asymmetric Pitx2 expression in the left lateral plate of Pitx2(Delta)(ASE/)(Delta)(ASE) embryos was not sufficient to restore normal organogenesis, suggesting that continuous expression of Pitx2 in the lineage of the left lateral plate is required for situs-specific organogenesis.


[Clinical evaluation of a bleomycin-group antitumor antibiotic bleomycetin].
The efficacy of bleomycetin or bleomycin A5 was studied in 128 patients with different malignant neoplasms. The antibiotic was used as a systemic or intracavitary chemotherapeutic agent. Bleomycetin was effective in 75-80, 81.8, 58.3, 70 and 50 per cent of the cases with disseminated derminogenic tumor of the testicle, squamous cell carcinoma of the head and neck, cancer of the penis, carcinoma of the skin and lymphogranulomatosis, respectively. When used intracavitarily the drug was effective in 41.2 per cent of the patients with cancer of the ovaries and lungs, teratoblastoma of the ovaries, cancer of the mammary gland and sarcoma of the soft tissues. Hyperthermia and focal hyperkeratosis as the adverse reactions were observed in 40.6 and 5.4 per cent of the patients, respectively. No toxicity with respect to the lungs was registered.


[Insertion of Trypanosoma cruzi DNA in the genome of mammal host cell through infection].
Trypanosoma cruzi chromatin was observed in macrophage chromosome spreads obtained at different periods of infections in BALB/c mice. Immunofluorescent studies showed that genetic materials associated with the chromosomes were T. cruzi products. In situ hybridization showed the protozoon DNA insertion in the host cell genome. In addition, selective 3H-DNA insertion in chromosomes 3, 6 and 11 was observed, which suggested that transxenogene rearrangement may take place in T. cruzi infections of mammals.


Determining the Optimum Bar Velocity in the Barbell Hip Thrust Exercise.
To identify the bar velocities that optimize power output in the barbell hip thrust exercise. A total of 40 athletes from 2 sports disciplines (30 track-and-field sprinters and jumpers and 10 rugby union players) participated in this study. Maximum bar-power outputs and their respective bar velocities were assessed in the barbell hip thrust exercise. Athletes were divided, using a median split analysis, into 2 groups according to their bar-power outputs in the barbell hip thrust exercise ("higher" and "lower" power groups). The magnitude-based inferences method was used to analyze the differences between groups in the power and velocity outcomes. To assess the precision of the bar velocities for determining the maximum power values, the coefficient of variation (CV%) was also calculated. Athletes achieved the maximum power outputs at a mean velocity, mean propulsive velocity, and peak velocity of 0.92 (0.04) m·s-1 (CV: 4.1%), 1.02 (0.05) m·s-1 (CV: 4.4%), and 1.72 (0.14) m·s-1 (CV: 8.4%), respectively. No meaningful differences were observed in the optimum bar velocities between higher and lower power groups. Independent of the athletes' power output and bar-velocity variable, the optimum power loads frequently occur at very close bar velocities.


Tumor growth and angiogenesis is impaired in CIB1 knockout mice.
Pathological angiogenesis contributes to various ocular, malignant, and inflammatory disorders, emphasizing the need to understand this process more precisely on a molecular level. Previously we found that CIB1, a 22 kDa regulatory protein, plays a critical role in endothelial cell function, angiogenic growth factor-mediated cellular functions, PAK1 activation, MMP-2 expression, and in vivo ischemia-induced angiogenesis. Since pathological angiogenesis is highly dependent on many of these same processes, we hypothesized that CIB1 may also regulate tumor-induced angiogenesis. To test this hypothesis, we allografted either murine B16 melanoma or Lewis lung carcinoma cells into WT and CIB1-KO mice, and monitored tumor growth, morphology, histology, and intra-tumoral microvessel density. Allografted melanoma tumors that developed in CIB1-KO mice were smaller in volume, had a distinct necrotic appearance, and had significantly less intra-tumoral microvessel density. Similarly, allografted Lewis lung carcinoma tumors in CIB1-KO mice were smaller in volume and mass, and appeared to have decreased perfusion. Intra-tumoral hemorrhage, necrosis, and perivascular fibrosis were also increased in tumors that developed in CIB1-KO mice. These findings suggest that, in addition to its other functions, CIB1 plays a critical role in facilitating tumor growth and tumor-induced angiogenesis.


Interaction of arginine, lysine, and guanidine with surface residues of lysozyme: implication to protein stability.
Additives are widely used to suppress aggregation of therapeutic proteins. However, the molecular mechanisms of effect of additives to stabilize proteins are still unclear. To understand this, we herein perform molecular dynamics simulations of lysozyme in the presence of three commonly used additives: arginine, lysine, and guanidine. These additives have different effects on stability of proteins and have different structures with some similarities; arginine and lysine have aliphatic side chain, while arginine has a guanidinium group. We analyze atomic contact frequencies to study the interactions of the additives with individual residues of lysozyme. Contact coefficient, quantified from contact frequencies, is helpful in analyzing the interactions with the guanidine groups as well as aliphatic side chains of arginine and lysine. Strong preference for contacts to the additives (over water) is seen for the acidic followed by polar and the aromatic residues. Further analysis suggests that the hydration layer around the protein surface is depleted more in the presence of arginine, followed by lysine and guanidine. Molecular dynamics simulations also reveal that the internal dynamics of protein, as indicated by the lifetimes of the hydrogen bonds within the protein, changes depending on the additives. Particularly, we note that the side-chain hydrogen-bonding patterns within the protein differ with the additives, with several side-chain hydrogen bonds missing in the presence of guanidine. These results collectively indicate that the aliphatic chain of arginine and lysine plays a critical role in the stabilization of the protein.


Magnetic resonance imaging in a patient with nitrous oxide-induced subacute combined degeneration of the spinal cord.
Chronic nitrous oxide use can lead to neurological findings that are clinically and radiographically identical to those found in patients with pernicious anemia, specifically subacute combined degeneration of the spinal cord and peripheral neuropathy. A 22-year-old man presented with lower extremity weakness and ataxia in the setting of inhaling 250 nitrous oxide cartridges two to three times weekly for two years. Magnetic resonance imaging showed T2 hyperenhancement of the dorsal columns of the cervical spine from the first to the sixth vertebrae, which helped to establish a diagnosis of nitrous oxide-induced subacute combined degeneration of the spinal cord. Chronic nitrous oxide use should be included in the differential diagnosis of any patient with otherwise unexplained neurological complaints that localize to the dorsal columns and has the changes on magnetic resonance imaging described here.


Gemcitabine sensitivity-related mRNA expression in endoscopic ultrasound-guided fine-needle aspiration biopsy of unresectable pancreatic cancer.
The aim of this study was to determine a predictive indicator of gemcitabine (GEM) efficacy in unresectable pancreatic cancer using tissue obtained by endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNA). mRNAs extracted from 35 pancreatic tubular adenocarcinoma tissues obtained by EUS-FNA before GEM-treatment were studied. mRNAs were amplified and applied to a Focused DNA Array, which was restricted to well-known genes, including GEM sensitivity-related genes, deoxycytidine kinase (dCK), human equilibrative nucleoside transporter 1 (hENT1), hENT2, dCMP deaminase, cytidine deaminase, 5'-nucleotidase, ribonucleotide reductase 1 (RRM1) and RRM2. mRNA levels were classified into high and low expression based on a cut-off value defined as the average expression of 35 samples. These 35 patients were divided into the following two groups. Patients with partial response and those with stable disease whose tumor markers decreased by 50% or more were classified as the effective group. The rest of patients were classified as the non-effective group. The relationship between GEM efficacy and mRNA expression was then examined by chi-squared test. Among these GEM sensitivity-related genes, dCK alone showed a significant correlation with GEM efficacy. Eight of 12 patients in the effective group had high dCK expression, whereas 16 of 23 patients in non-effective group had low dCK expressions (P = 0.0398). dCK mRNA expression is a candidate indicator for GEM efficacy in unresectable pancreatic cancer. Quantitative mRNA measurements of dCK using EUS-FNA samples are necessary for definitive conclusions.


Dysregulation of the proteasome increases the toxicity of ALS-linked mutant SOD1.
A hallmark of protein conformational disease, exemplified by neurodegenerative disorders, is the expression of misfolded and aggregated proteins. The relationship between protein aggregation and cellular toxicity is complex, and various models of experimental pathophysiology have often yielded conflicting or controversial results. In this study, we examined the biophysical properties of amyotrophic lateral sclerosis (ALS)-linked mutations of Cu/Zn superoxide dismutase 1 (SOD1) expressed in human tissue culture cells. Fluorescence correlation spectroscopy (FCS) and Förster resonance energy transfer (FRET) analyses revealed that changes in proteasome activity affected both the expression of FCS- and FRET-detected oligomers and cellular toxicity. Under normal conditions, highly aggregation-prone mutant SOD1 exhibited very little toxicity. However, when the activity of the proteasome was transiently inhibited, only upon recovery did we observe the appearance of ordered soluble oligomers, which were closely correlated with cellular toxicity. These results shed light on the importance of balance in proteostasis and suggest that transient shifts of activity in the cellular machinery can alter the course of protein conformational transitions and dysregulate modulation of proteasome activity. In neurodegenerative disorders including ALS, such changes may be a risk factor for pathogenesis.


Electrochemical principles for active control of liquids on submillimeter scales
Electrochemical methods were combined with redox-active surfactants to actively control the motions and positions of aqueous and organic liquids on millimeter and smaller scales. Surfactant species generated at one electrode and consumed at another were used to manipulate the magnitude and direction of spatial gradients in surface tension and guide droplets of organic liquids through simple fluidic networks. Solid microparticles could be transported across unconfined surfaces. Electrochemical control of the position of surface-active species within aqueous films of liquid supported on homogeneous surfaces was used to direct these films into periodic arrays of droplets with deterministic shapes and sizes.


[Construction of lentiviral vector pTK208-mVEGF-IRES-DsRed contains dsred and mVEGF and its expression in 293T cells].
To construct red fluorescent protein (DsRed) and mouse vascular endothelial growth factor (mVEGF) lentiviral expression vector in order to obtain the DsRed and VEGF expression in 293T cells. The mVEGF was amplified from the lungs of fetal mouse by RT-PCR. The IRES-DsRed was obtained from plasmid pLV-tTRKRAB-red which contains IRES-DsRed sequence by PCR. Then they were cloned into lentiviral expression vector pTK-208. Human embryonic kidney 293T cells were co-transfected with the three plasmids by lipofectamine, and the expression of DsRed was examined under fluorescent microscope 24 h and 48 h after transfection. 48 h and 72 h later the viral supernatant was used to infect 293T cells, the expression of DsRed was examined under fluorescent microscope and Western blot assays was used to examine the expression of mVEGF in cells and the culture medium. The recombinant lentiviral vector pTK208-mVEGF-IRES-DsRed was constructed, the virus titres were above 10(6) PFU/mL in the supernatant. After infection, DsRed and mVEGF proteins were successfully expressed in 293T cells. The lentiviral vector pTK208-mVEGF-IRES-DsRed contains DsRed and mVEGF were successfully constructed and expressed in 293T cells, it could provide a basis for further researches on the pathophysiological mechanism of VEGF and lay the foundation of gene therapy.


Coronal plane socket stability during gait in persons with transfemoral amputation: Pilot study.
Little research describes which transfemoral socket design features are important for coronal plane stability, socket comfort, and gait. Our study objectives were to (1) relate socket comfort during gait to a rank order of changes in ischial containment (IC) and tissue loading and (2) compare socket comfort during gait when tissue loading and IC were systematically manipulated. Six randomly assigned socket conditions (IC and tissue compression) were assessed: (1) IC and high, (2) IC and medium, (3) IC and low, (4) no IC and high, (5) no IC and medium, and (6) no IC and low. For the six subjects in this study, there was a strong, negative relationship between comfort and changes in IC and tissue loading (rho = -0.89). With the ischium contained, tissue loading did not influence socket comfort (p = 0.47). With no IC, the socket was equally comfortable with high tissue loading (p = 0.36) but the medium (p = 0.04) and low (p = 0.02) tissue loading conditions decreased comfort significantly. Coronal plane hip moments, lateral trunk lean, step width, and walking speed were invariant to changes in IC and/or tissue loading. Our results suggest that in an IC socket, medial tissue loading mattered little in terms of comfort. Sockets without IC required high tissue loading to be as comfortable as those with IC, while suboptimal tissue loading compromised comfort.


High accuracy modeling of photodiode quantum efficiency.
We propose a new silicon photodiode model optimized for high-accuracy measurement usage. The new model differs from previous models in that the contribution to the quantum efficiency from the diode front region is described by an integral transform of the equilibrium minority carrier concentration. This description is accurate as long as the recombination of excess minority carriers in the front region occurs only at the front surface and the diode is operating linearly.


Isolated conjunctival lichen planus: a diagnostic challenge.
Lichen planus is a common inflammatory autoimmune condition of unknown etiology that commonly affects the skin and mucous membranes. Isolated ocular lichen planus is an extremely rare presentation that most commonly involves the eyelids, conjunctiva, and cornea, leading to severe scarring, and is clinically indistinguishable from other causes of cicatricial conjunctivitis. A 79-year-old man complained of a chronic keratoconjunctivitis refractory to multiple topical treatments. Slit-lamp examination revealed diffuse bilateral conjunctival hyperemia, subepithelial fibrosis, and symblepharon, with a marked shortening of the lower conjunctival fornix. There were no other skin or mucosal lesions. Hematoxylin-eosin staining revealed acanthosis, focal thickening of the basement membrane, and a dense subepithelial mononuclear infiltrate. Direct immunofluorescence demonstrated a linear shaggy fibrinogen deposition along the basement membrane, suggestive of lichen planus. Ultrastructural examination revealed a marked widening of the epithelium-lamina propria interphase, with prominent fragmentation, reduplication, and reticulation of the lamina densa of the basement membrane. The patient was successfully treated with systemic immunosuppressive agents. Isolated conjunctival lichen planus is an exceptional and severe cause of cicatricial conjunctivitis. Distinguishing this unusual presentation from other inflammatory diseases with conjunctival involvement is crucial to initiate an appropriate therapy early to avoid irreversible damage of the visual function.


Climate, urbanization, and species traits interactively drive flowering duration.
A wave of green leaves and multi-colored flowers advances from low to high latitudes each spring. However, little is known about how flowering offset (i.e., ending of flowering) and duration of populations of the same species vary along environmental gradients. Understanding these patterns is critical for predicting the effects of future climate and land-use change on plants, pollinators, and herbivores. Here, we investigated potential climatic and landscape drivers of flowering onset, offset, and duration of 52 plant species with varying key traits. We generated phenology estimates using >270,000 community-science photographs and a novel presence-only phenometric estimation method. We found longer flowering durations in warmer areas, which is more obvious for summer-blooming species compared to spring-bloomers driven by their strongly differing offset dynamics. We also found that higher human population density and higher annual precipitation are associated with delayed flowering offset and extended flowering duration. Finally, offset of woody perennials was more sensitive than herbaceous species to both climate and urbanization drivers. Empirical forecast models suggested that flowering durations will be longer in 2030 and 2050 under representative concentration pathway (RCP) 8.5, especially for summer-blooming species. Our study provides critical insight into drivers of key flowering phenophases and confirms that Hopkins' Bioclimatic Law also applies to flowering durations for summer-blooming species and herbaceous spring-blooming species.


Phonetic discrimination, phonological awareness, and pre-literacy skills in Spanish-English dual language preschoolers.
The current study explores variation in phonemic representation among Spanish-English dual language learners (DLLs, n = 60) who were dominant in English or in Spanish. Children were given a phonetic discrimination task with speech sounds that: 1) occur in English and Spanish, 2) are exclusive to English, and 3) are exclusive to Russian, during Fall (age m = 57 months) and Spring (age m = 62 months, n = 42). In Fall, English-dominant DLLs discriminated more accurately than Spanish-dominant DLLs between English-Spanish phones and English-exclusive phones. Both groups discriminated Russian phones at or close to chance. In Spring, however, groups no longer differed in discriminating English-exclusive phones and both groups discriminated Russian phones above chance. Additionally, joint English-Spanish and English-exclusive phonetic discrimination predicted children's phonological awareness in both groups. Results demonstrate plasticity in early childhood through diverse language exposure and suggest that phonemic representation begins to emerge driven by lexical restructuring.


[Rotavirus prevalence and genotype distribution in children with acute gastroenteritis in Adana province].
Rotavirus is the most common agent of acute gastroenteritis in children under five years of age and still a major cause of mortality and morbidity mainly in devoloping countries. The aim of this study was to determine the circulating rotavirus genotypes in children less than five years of age with acute gastroenteritis in Adana province, before the introduction of the rotavirus vaccine into the national immunization programme. Fecal specimens were collected from children enrolled between December 2012 and November 2016 and rotavirus antigen was searched by a rapid immunochromatographic test method. The antigen positive samples were G and Pgenotyped by hemi-nested type-specific multiplex reverse-transcriptase polymerase chain reaction (RT-PCR). Rotavirus antigen positive fecal specimens from 201 patients of 107 (53.3%) boys and 94 (46.8%) girls were included in this study. Rotavirus infection was most frequently detected in patients with gastroenteritis between the ages of 12 and 23 months. It has been determined that 45% of rotavirus gastroenteritis was determined in children under two years of age. Rotavirus gastroenteritis was more frequent between September and April, but the highest incidence was detected in January. Of 201 rotavirus antigen-positive stool specimens, 167 were PCR positive. Five common G (G1, G2, G3, G4 and G9) and two common P genotypes (P[8] and P[4]) were determined as 91.6% and 96.4% of the isolates, respectively. The most prevalent G and P genotype combination was G1P8 with a frequency of 29.9%, followed by 21% G9P8 About 78% of rotavirus isolates were classified into five common G-P combination genotypes, including G1P8, G9P8 G3P8, G2P4, and G2P8. During the study period, there were significant changes in the distribution of rotavirus genotype. The prevalence of G9P8 genotype sharply decreased from 40% to 8.1% and the prevalence of G1P8 genotype increased from 21.3% to 48.6%. Although no G3P8 isolates were detected in the first two years of the study, this genotype was detected in 18.5% and 13.5% of the samples in 2015 and 2016, respectively. There were no relationship between the age onset of the disease, sex, dehydration development risk and rotavirus genotype. The results from this study indicated that the predominant circulating strains of rotavirus determined in clinical specimens were G1P8 and G9P8. Significant changes were observed in the prevalence of the most common rotavirus isolates over the years. Rotavirus surveillance is important to determine the circulating strains of rotavirus to assess vaccine effectiveness, to monitor vaccine failures, and to detect potential emerging strains.


Nanocellulose/bioactive glass cryogels as scaffolds for bone regeneration.
A major challenge exists in the preparation of scaffolds for bone regeneration, namely, achieving simultaneously bioactivity, biocompatibility, mechanical performance and simple manufacturing. Here, cellulose nanofibrils (CNF) are introduced for the preparation of scaffolds taking advantage of their biocompatibility and ability to form strong 3D porous networks from aqueous suspensions. CNF are made bioactive for bone formation through a simple and scalable strategy that achieves highly interconnected 3D networks. The resultant materials optimally combine morphological and mechanical features and facilitate hydroxyapatite formation while releasing essential ions for in vivo bone repair. The porosity and roughness of the scaffolds favor several cell functions while the ions act in the expression of genes associated with cell differentiation. Ion release is found critical to enhance the production of the bone morphogenetic protein 2 (BMP-2) from cells within the fractured area, thus accelerating the in vivo bone repair. Systemic biocompatibility indicates no negative effects on vital organs such as the liver and kidneys. The results pave the way towards a facile preparation of advanced, high performance CNF-based scaffolds for bone tissue engineering.


A simulation study of motion artefacts in computed tomography.
In order to increase understanding of artefacts caused by patient motion in computed tomography, rigid translational and rotational motions in the plane of reconstruction have been considered. Motion artefacts were simulated on a digital computer and demonstrated in actual CT scans of a phantom. The types of motion artefacts observed included streaking, boundary splitting and subtle distortions of shape. A method for removing artefacts from a CT image without rescanning the patient is discussed, and some examples of possible improvement are shown using the simulation model. It is assumed that perfect knowledge of patient motion is known since this represents the theoretical limit on possible improvement to the degraded image. A qualitative discussion of streaking, the most common type of motion artefact, is also included.


Difficult cases in heart failure: amiodarone lung injury: another heart failure mimic?
Careful thought must be given to the development of bystander pathology that could mimic worsening of heart failure. Recent trials with patients receiving amiodarone record a low rate of amiodarone pulmonary toxicity of 1.6%. Bronchoalveolar lavage in amiodarone toxicity demonstrates an absolute and relative lymphocytic alveolitis, suggesting hypersensitivity, but this finding is neither sensitive nor specific. Recently, KL-6, a mucin-like high molecular weight glycoprotein secreted by proliferating type II alveolar pneumocytes, has been identified as a potential marker of interstitial pneumonitis. A high index of suspicion combined with rapid exclusion of common confounding mimics can help in establishing the diagnosis of amiodarone lung toxicity.


Vancomycin Dosing in Obese Patients: Special Considerations and Novel Dosing Strategies.
To review the literature regarding vancomycin pharmacokinetics in obese patients and strategies used to improve dosing in this population. PubMed, EMBASE (1974 to November 2017), and Google Scholar searches were conducted using the search terms vancomycin, obese, obesity, pharmacokinetics, strategy, and dosing. Additional articles were selected from reference lists of selected studies. Included articles were those published in English with a primary focus on vancomycin pharmacokinetic parameters in obese patients and practical vancomycin dosing strategies, clinical experiences, or challenges of dosing vancomycin in this population. Volume of distribution and clearance are the pharmacokinetic parameters that most often affect vancomycin dosing in obese patients; both are increased in this population. Challenges with dosing in obese patients include inconsistent and inadequate dosing, observations that the obese population may not be homogeneous, and reports of an increased likelihood of supratherapeutic trough concentrations. Investigators have revised and developed dosing and monitoring protocols to address these challenges. These approaches improved target trough attainment to varying degrees. Some of the vancomycin dosing approaches provided promising results in obese patients, but there were notable differences in methods used to develop these approaches, and sample sizes were small. Although some approaches can be considered for validation in individual institutions, further research is warranted. This may include validating approaches in larger populations with narrower obesity severity ranges, investigating target attainment in indication-specific target ranges, and evaluating the impact of different dosing weights and methods of creatinine clearance calculation.


USP32 is an active, membrane-bound ubiquitin protease overexpressed in breast cancers.
USP32, on chromosomal band 17q23.1-17q23.2, is a highly conserved but uncharacterized gene that gave rise during evolution to a well-known hominoid-specific proto-oncogene, USP6. We investigated the expression profile of USP32 in human tissues and examined its functions to gain insight into this novel member of the well-conserved ubiquitination system. We detected ubiquitous USP32 expression across tissues and confirmed the predicted deubiquitination function owing to the presence of conserved peptidase signature aspargine, cysteine, histidine, and aspartic acid domains of ubiquitin-specific proteases. A Golgi localization of GFP-fused USP32 was detected by fluorescent protection assay and BODIPY-TR staining. In addition, stable silencing of USP32 caused a significant decrease in the proliferation and migration rate of cells. Based on these and the fact that USP32 maps to 17q23, which is commonly amplified in breast cancers, we analyzed USP32 expression in breast cancer cells. We detected high expression of USP32 in 50% (9 of 18) of breast cancer cell lines and 22% (9 of 41) of primary breast tumors compared to mammary epithelial cells. In summary, we report the preliminary characterization of this novel deubiquitinating enzyme on 17q23 and demonstrate its functional role in the ubiquitin system and its potential involvement in tumorigenesis.


Inequities in the incidence and safety of abortion in Nigeria.
We know little about the frequency, correlates and conditions under which women induce abortions in Nigeria. This study seeks to estimate the 1-year induced abortion incidence and proportion of abortions that are unsafe overall and by women's background characteristics using direct and indirect methodologies. Data for this study come from a population-based, nationally representative survey of reproductive age women (15-49) in Nigeria. Interviewers asked women to report on the abortion experiences of their closest female confidante and themselves. We adjusted for potential biases in the confidante data. Analyses include estimation of 1-year induced abortion incidence and unsafe abortion, as well as bivariate and multivariate assessment of their correlates. A total of 11 106 women of reproductive age completed the female survey; they reported on 5772 confidantes. The 1-year abortion incidence for respondents was 29.0 (95% CI 23.3 to 34.8) per 1000 women aged 15-49 while the confidante incidence was 45.8 (95% CI 41.0-50.6). The respondent and confidante abortion incidences revealed similar correlates, with women in their 20s, women with secondary or higher education and women in urban areas being the most likely to have had an abortion in the prior year. The majority of respondent and confidante abortions were the most unsafe (63.4% and 68.6%, respectively). Women aged 15-19, women who had never attended school and the poorest women were significantly more likely to have had the most unsafe abortions. Results indicate that abortion in Nigeria is a public health concern and an issue of social inequity. Efforts to expand the legal conditions for abortion in Nigeria are critical. Simultaneously, efforts to increase awareness of the availability of medication abortion drugs to more safely self-induce can help mitigate the toll of unsafe abortion-related morbidity and mortality.


The effect of 4-weeks exposure to music on social bonding between rats.
Interpersonal synchronization of movement induced by music is believed to facilitate social bonding between human beings, but it is unknown whether it also works in animals. We allowed rats to interact and develop social bonding with a specific subject for four weeks under one of the three acoustic conditions: playback of K.448 at its original tempo, playback at its double-tempo, and silence. The strength of social bonding between each pair of rats was then measured. The results showed an increase in preference for rats that had interacted under the original tempo playback compared to the other acoustic conditions. Considering that rats move in synchrony with the beat more robustly and consistently between subjects under the original tempo playback than under the double-tempo playback, this result suggests that motor synchronization between subjects through music may facilitate social bonding between rats.


Whole genome transcriptomics reveals global effects including up-regulation of Francisella pathogenicity island gene expression during active stringent response in the highly virulent Francisella tularensis subsp. tularensis SCHU S4.
During conditions of nutrient limitation bacteria undergo a series of global gene expression changes to survive conditions of amino acid and fatty acid starvation. Rapid reallocation of cellular resources is brought about by gene expression changes coordinated by the signalling nucleotides' guanosine tetraphosphate or pentaphosphate, collectively termed (p)ppGpp and is known as the stringent response. The stringent response has been implicated in bacterial virulence, with elevated (p)ppGpp levels being associated with increased virulence gene expression. This has been observed in the highly pathogenic Francisella tularensis sub spp. tularensis SCHU S4, the causative agent of tularaemia. Here, we aimed to artificially induce the stringent response by culturing F. tularensis in the presence of the amino acid analogue l-serine hydroxamate. Serine hydroxamate competitively inhibits tRNAser aminoacylation, causing an accumulation of uncharged tRNA. The uncharged tRNA enters the A site on the translating bacterial ribosome and causes ribosome stalling, in turn stimulating the production of (p)ppGpp and activation of the stringent response. Using the essential virulence gene iglC, which is encoded on the Francisella pathogenicity island (FPI) as a marker of active stringent response, we optimized the culture conditions required for the investigation of virulence gene expression under conditions of nutrient limitation. We subsequently used whole genome RNA-seq to show how F. tularensis alters gene expression on a global scale during active stringent response. Key findings included up-regulation of genes involved in virulence, stress responses and metabolism, and down-regulation of genes involved in metabolite transport and cell division. F. tularensis is a highly virulent intracellular pathogen capable of causing debilitating or fatal disease at extremely low infectious doses. However, virulence mechanisms are still poorly understood. The stringent response is widely recognized as a diverse and complex bacterial stress response implicated in virulence. This work describes the global gene expression profile of F. tularensis SCHU S4 under active stringent response for the first time. Herein we provide evidence for an association of active stringent response with FPI virulence gene expression. Our results further the understanding of the molecular basis of virulence and regulation thereof in F. tularensis. These results also support research into genes involved in (p)ppGpp production and polyphosphate biosynthesis and their applicability as targets for novel antimicrobials.


Bridging the gap: incorporating exercise evidence into clinical practice in breast cancer care.
To determine the feasibility (recruitment, retention, and adherence rates) of implementing a multi-dimensional knowledge translation (KT) intervention designed for women with breast cancer, and to explore preliminary estimates of effects of the implementation strategy on exercise level, exercise knowledge and behavior, health-related quality of life, and overall health status among breast cancer survivors. Design: Implementation Trial. Community-dwelling women, over 18 years of age, currently undergoing chemotherapy for stage 1-3 BRCA. Randomization and Blinding: A blinded researcher randomized participants on a record-by-record basis to the intervention or control group. A researcher blinded to the group allocation of the participants conducted the statistical analysis. Intervention Group: Eight sessions of moderate intensity aerobic exercise along with eight self-management modules were delivered during chemotherapy within the cancer institution. Usual care. Feasibility of implementation strategy measured through recruitment, retention, and adherence rates. Twenty-nine women were screened for this study. Twenty-seven met inclusion criteria and twenty-six participants were enrolled in the study. The implementation strategy was determined to be feasible and had a recruitment rate of 96%, retention rate of 100%, and adherence rate of 89%. The intervention group had significantly higher exercise levels (mean difference = 25.38, 95%CI = (9.35, 41.42)) post-intervention compared with the control group. No adverse events were reported. The implementation of this KT intervention is feasible and demonstrates preliminary effects for secondary outcomes for women with breast cancer during chemotherapy. Findings support the implementation of this intervention in a multi-center trial. NCT03087461.


Neurofibromin (Nf1) is required for skeletal muscle development.
Neurofibromatosis type 1 (NF1) is a multi-system disease caused by mutations in the NF1 gene encoding a Ras-GAP protein, neurofibromin, which negatively regulates Ras signaling. Besides neuroectodermal malformations and tumors, the skeletal system is often affected (e.g. scoliosis and long bone dysplasia) demonstrating the importance of neurofibromin for development and maintenance of the musculoskeletal system. Here, we focus on the role of neurofibromin in skeletal muscle development. Nf1 gene inactivation in the early limb bud mesenchyme using Prx1-cre (Nf1(Prx1)) resulted in muscle dystrophy characterized by fibrosis, reduced number of muscle fibers and reduced muscle force. This was caused by an early defect in myogenesis affecting the terminal differentiation of myoblasts between E12.5 and E14.5. In parallel, the muscle connective tissue cells exhibited increased proliferation at E14.5 and an increase in the amount of connective tissue as early as E16.5. These changes were accompanied by excessive mitogen-activated protein kinase pathway activation. Satellite cells isolated from Nf1(Prx1) mice showed normal self-renewal, but their differentiation was impaired as indicated by diminished myotube formation. Our results demonstrate a requirement of neurofibromin for muscle formation and maintenance. This previously unrecognized function of neurofibromin may contribute to the musculoskeletal problems in NF1 patients.


Home blood pressure measurement in elderly patients with cognitive impairment: comparison of agreement between relative-measured blood pressure and automated blood pressure measurement.
Home blood pressure measurement (HBPM) is recommended by guidelines for hypertension management. However, this method might be difficult to use in elderly individuals with cognitive disorders. Our aim was to assess the agreement and the feasibility of HBPM by a relative as compared with 24-h ambulatory blood pressure monitoring (ABPM) in elderly patients with dementia. Sixty outpatients with dementia aged 75 years and older with office hypertension (≥140/90 mmHg) were subjected successively to HBPM by a trained relative and 24-h ABPM. The order of the two methods was randomized. Current guidelines' thresholds for the diagnosis of hypertension were used. The mean (SD) age of the patients was 80.8 (6.1) years (55% women) and the mean (SD) mini-mental state examination score was 20.1 (6.9). The feasibility of relative-HBPM was very high, with a 97% success rate (defined by ≥12/18 measurements reported). The blood pressure measurements were highly correlated between the two methods (r=0.75 and 0.64 for systolic blood pressure and diastolic blood pressure, respectively; P<0.001 for both). The agreement between the methods for the diagnosis of sustained hypertension and white-coat hypertension was excellent (overall agreement, 92%; κ coefficient, 0.81; 95% CI, 0.61-0.93). Similar results were found for daytime-ABPM. In cognitively impaired elderly patients, HBPM by a relative using an automated device was a good alternative to 24-h ABPM.


Detection of spectral signatures in multispectral MR images for classification.
This paper presents a new spectral signature detection approach to magnetic resonance (MR) image classification. It is called constrained energy minimization (CEM) method, which is derived from the minimum variance distortionless response in passive sensor array processing. It considers a bank of spectral channels as an array of sensors where each spectral channel represents a sensor and object spectral signature in multispectral MR images are viewed as signals impinging upon the array. The strength of the CEM lies on its ability in detection of spectral signatures of interest without knowing image background. The detected spectral signatures are then used for classification. The CEM makes use of a finite impulse response (FIR) filter to linearly constrain a desired object while minimizing interfering effects caused by other unknown signal sources. Unlike most spatial-based classification techniques, the proposed CEM takes advantage of spectral characteristics to achieve object detection and classification. A series of experiments is conducted and compared with the commonly used c-means method for performance evaluation. The results show that the CEM method is a promising and effective spectral technique for MR image classification.


A 23-watt single-frequency vertical-external-cavity surface-emitting laser.
We report on a single-frequency semiconductor disk laser which generates 23.6 W output power in continuous wave operation, at a wavelength of 1013 nm. The high output power is a result of optimizing the chip design, thermal management and the cavity configuration. By applying passive stabilization techniques, the free-running linewidth is measured to be 407 kHz for a sampling time of 1 ms, while undercutting 100 kHz in the microsecond domain.


Mild/asymptomatic COVID-19 in unvaccinated pregnant mothers impairs neonatal immune responses.
Maternal SARS-CoV-2 infection triggers placental inflammation and alters cord blood immune cell composition. However, most studies focus on outcomes of severe maternal infection. Therefore, we analyzed cord blood and chorionic villi from newborns of unvaccinated mothers who experienced mild/asymptomatic SARS-CoV-2 infection during pregnancy. We investigated immune cell rewiring using flow cytometry, single-cell RNA sequencing, and functional readouts using ex vivo stimulation with TLR agonists and pathogens. Maternal infection was associated with increased frequency of memory T and B cells and nonclassical monocytes in cord blood. Ex vivo T and B cell responses to stimulation were attenuated, suggesting a tolerogenic state. Maladaptive responses were also observed in cord blood monocytes, where antiviral responses were dampened but responses to bacterial TLRs were increased. Maternal infection was also associated with expansion and activation of placental Hofbauer cells, secreting elevated levels of myeloid cell-recruiting chemokines. Moreover, we reported increased activation of maternally derived monocytes/macrophages in the fetal placenta that were transcriptionally primed for antiviral responses. Our data indicate that even in the absence of vertical transmission or symptoms in the neonate, mild/asymptomatic maternal COVID-19 altered the transcriptional and functional state in fetal immune cells in circulation and in the placenta.


Genetic vulnerability to Crohn's disease reveals a spatially resolved epithelial restitution program.
Effective tissue repair requires coordinated intercellular communication to sense damage, remodel the tissue, and restore function. Here, we dissected the healing response in the intestinal mucosa by mapping intercellular communication at single-cell resolution and integrating with spatial transcriptomics. We demonstrated that a risk variant for Crohn's disease, hepatocyte growth factor activator (HGFAC) Arg509His (R509H), disrupted a damage-sensing pathway connecting the coagulation cascade to growth factors that drive the differentiation of wound-associated epithelial (WAE) cells and production of a localized retinoic acid (RA) gradient to promote fibroblast-mediated tissue remodeling. Specifically, we showed that HGFAC R509H was activated by thrombin protease activity but exhibited impaired proteolytic activation of the growth factor macrophage-stimulating protein (MSP). In Hgfac R509H mice, reduced MSP activation in response to wounding of the colon resulted in impaired WAE cell induction and delayed healing. Through integration of single-cell transcriptomics and spatial transcriptomics, we demonstrated that WAE cells generated RA in a spatially restricted region of the wound site and that mucosal fibroblasts responded to this signal by producing extracellular matrix and growth factors. We further dissected this WAE cell-fibroblast signaling circuit in vitro using a genetically tractable organoid coculture model. Collectively, these studies exploited a genetic perturbation associated with human disease to disrupt a fundamental biological process and then reconstructed a spatially resolved mechanistic model of tissue healing.


A novel poxvirus lethal to red squirrels (Sciurus vulgaris).
A parapoxvirus has been implicated in the decline of the red squirrel in the United Kingdom. Virus was isolated from an outbreak of lethal disease in red squirrels in the north-east of England. Experimental infection of captive-bred red squirrels confirmed that this virus was the cause of the severe skin lesions observed. Electron microscopic examination of the virus showed that it had a morphology typical of parapoxviruses whilst preliminary sequence data suggested a genomic G+C composition of approximately 66 %, again similar to that found in other parapoxviruses. However Southern hybridization analysis failed to detect three known parapoxvirus genes, two of which have been found so far only in the genus parapoxvirus. Comparative sequence analysis of two other genes, conserved across the eight recognized chordopoxvirus genera, suggests that the squirrel virus represents a previously unrecognized genus of the chordopoxvirus.


Extended pelvic lymph node dissection in robotic-assisted radical prostatectomy: surgical technique and initial experience.
To describe, and show in the accompanying video segments, a technique for extended pelvic lymph node dissection (ePLND) in robotic-assisted radical prostatectomy (RARP) and report our clinicopathologic and perioperative outcomes. The extent of pelvic lymphadenectomy during radical prostatectomy has not been standardized. However, evidence demonstrates that an ePLND yields a greater number of positive nodes. A total of 32 patients with clinically localized prostate cancer underwent RARP with ePLND by a single surgeon (J.C.) between January and August 2008. The template for the ePLND included the obturator, hypogastric, external iliac, and common iliac lymph nodes up to the bifurcation of the aorta. Systematic review and grading of adverse events were performed. The median number of lymph nodes retrieved was 18 (interquartile range [IQR] 12-28). Four patients (12.5%) had lymph node metastases. Of the 4 patients with lymph node metastases, 1 patient (25%) had the involved lymph node exclusively in the common iliac region. Median operative time for the ePLND was 72 minutes (IQR 66-86). Median hospital length of stay was 2.0 days (IQR 2.0-2.8). Graded complications included 13 grade 1 events and 1 grade 2 event, with 1 grade 1 event being considered related to ePLND. No clinically presenting lymphoceles or thrombotic events were encountered. An ePLND during RARP is technically feasible and appears to have minimal morbidity. It produces a high lymph node yield and may result in improved pathologic staging.


The PEST Model: A Quality Improvement Project to Reduce Self-extubation in the Intensive Care Unit.
Self-extubation is an adverse patient event that can lead to severe complications. Gaps in clinical practice from the lack of nursing awareness and decision making capacity have often resulted in cases of preventable self-extubation. Review of current evidence suggests that initiatives to support nursing clinical decision making can help prevent adverse patient events such as self-extubation. The aim of this study was to reduce the incidence of self-extubation by 50% in a cardiology intensive care unit over 1 year. A quality improvement project was undertaken with a PEST model of nursing care introduced from January 2017 to December 2017 in the cardiology intensive care unit to guide nursing staff to assess and render appropriate interventions along patient domains such as pain, endotracheal tube securement, sedation, and tie to prevent incidences of self-extubation. Incidences of self-extubation have reduced to 5 cases in 2017, reflecting a 50% improvement from 10 cases in 2016. Formalizing practice standards into an easy-to-remember mnemonics or framework can improve patient outcomes. Policy makers must be aware that initiatives to facilitate decision making can improve patient safety.


Isorenieratene biosynthesis in green sulfur bacteria requires the cooperative actions of two carotenoid cyclases.
The cyclization of lycopene to gamma- or beta-carotene is a major branch point in the biosynthesis of carotenoids in photosynthetic bacteria. Four families of carotenoid cyclases are known, and each family includes both mono- and dicyclases, which catalyze the formation of gamma- and beta-carotene, respectively. Green sulfur bacteria (GSB) synthesize aromatic carotenoids, of which the most commonly occurring types are the monocyclic chlorobactene and the dicyclic isorenieratene. Recently, the cruA gene, encoding a conserved hypothetical protein found in the genomes of all GSB and some cyanobacteria, was identified as a lycopene cyclase. Further genomic analyses have found that all available fully sequenced genomes of GSB encode an ortholog of cruA. Additionally, the genomes of all isorenieratene-producing species of GSB encode a cruA paralog, now named cruB. The cruA gene from the chlorobactene-producing GSB species Chlorobaculum tepidum and both cruA and cruB from the brown-colored, isorenieratene-producing GSB species Chlorobium phaeobacteroides strain DSM 266(T) were heterologously expressed in lycopene- and neurosporene-producing strains of Escherichia coli, and the cruB gene of Chlorobium clathratiforme strain DSM 5477(T) was also heterologously expressed in C. tepidum by inserting the gene at the bchU locus. The results show that CruA is probably a lycopene monocyclase in all GSB and that CruB is a gamma-carotene cyclase in isorenieratene-producing species. Consequently, the branch point for the synthesis of mono- and dicyclic carotenoids in GSB seems to be the modification of gamma-carotene, rather than the cyclization of lycopene as occurs in cyanobacteria.


[Erik Essen-Möller and the roots of multiaxial classification in psychiatry].
Against the background of the preparation of ICD-11 and DSM-V, the historical roots of a multiaxial diagnostic assessment in psychiatry are reviewed. The principles of such an approach are traced back to the Swedish psychiatrist Erik Essen-Möller who had proposed a distinction between aetiological and descriptive aspects in the classification of mental disorders. Furthermore, he suggested to break down the descriptive classification into the cross-sectional psychopathological picture and the clinical course. Nowadays, a multiaxial assessment is used in diagnostic systems such as DSM-IV. However, these current concepts differ considerably from Essen-Möller's suggestions. A return to the original approach of multiaxial diagnostic assessment comprising the axes "syndromes", "course types" and "aetiology" would be in line with current neurobiological findings and may provide a bridge between the traditional categorical diagnostic approach and dimensional models.


Agenda-setting for Canadian caregivers: using media analysis of the maternity leave benefit to inform the compassionate care benefit.
The Compassionate Care Benefit was implemented in Canada in 2004 to support employed informal caregivers, the majority of which we know are women given the gendered nature of caregiving. In order to examine how this policy might evolve over time, we examine the evolution of a similar employment insurance program, Canada's Maternity Leave Benefit. National media articles were reviewed (n = 2,698) and, based on explicit criteria, were analyzed using content analysis. Through the application of Kingdon's policy agenda-setting framework, the results define key recommendations for the Compassionate Care Benefit, as informed by the developmental trajectory of the Maternity Leave Benefit. Recommendations for revising the Compassionate Care Benefit are made.


Graded testosterone infusions distinguish gonadotropin negative-feedback responsiveness in Asian and white men--a Clinical Research Center study.
Recently, multicenter clinical trials to determine male contraceptive efficacy disclosed that testosterone-induced suppression of spermatogenesis to azoospermia occurred in about 90% of Asian but only 60-70% of white men. To test whether there are ethnic differences in the sensitivity of gonadotropin secretion to suppression by testosterone, we administered constant infusions of testosterone at 0, 7, 14, and 28 mg/1.7 m2 x 24 h i.v. for 48 h to 9 Asian and 8 white normal male volunteers (22-42 yr old). During the last 8 h of each infusion dose, 10-min frequent blood sampling was carried out for later LH and FSH measurements by sensitive fluoroimmunoassays. Analyses of LH secretory pulses showed that LH pulse width, height, area, and total area under the curve (LH concentration vs. time) were significantly more suppressed in Asians than in whites during the lowest infusion dose of testosterone. With increasing testosterone dose, the suppression of pulsatile LH secretion was not different in the two ethnic groups. In contrast to pulsatile LH secretion, the responsiveness of pulsatile FSH secretion to exogenous testosterone infusion was not different between the two ethnic groups. At baseline, Asian men had a significantly higher mean number of FSH pulses and mean incremental pulse heights than did white men. Serum inhibin B levels were not distinguishable in the two ethnic groups, but the FSH profiles were quantifiably more irregular (higher approximate entropy) in the Asian volunteers. Our data suggest that, compared with white men, Asian men respond earlier and with more marked suppression of pulsatile LH secretion to ramped testosterone infusions. The elevated basal serum FSH concentrations (and more irregular FSH release pattern) observed in Asian men may suggest a small relative decrease in spermatogenic reserve and/or gonadal negative feedback. Whether these differences contribute to the observed differences in suppression of spermatogenesis in Asians vs. non-Asians in male contraceptive studies is not yet known.


Evaluation of Outcome Following Coronectomy for the Management of Mandibular Third Molars in Close Proximity to Inferior Alveolar Nerve.
Iatrogenic damage to Inferior Alveolar Nerve (IAN) is a significant risk factor following prophylactic or therapeutic removal of impacted mandibular third molar. The risk to IAN injury increases many fold, when the third molar root overlaps the nerve canal as identified by the radiographic imaging. Various methods like orthodontic assisted extraction, staged removal of tooth or coronectomy have been advocated to reduce the incidence of IAN injury in high risk cases with variable outcome. The aim of present study was to evaluate the fate of the root (resorbed, exfoliated, covered by bone) after coronectomy or intentional root retention of impacted mandibular 3(rd) molars in patients with high risk for inferior alveolar nerve damage as evaluated by the intra oral periapical radiograph. Twenty impacted mandibular third molar teeth, in 18 patients with high risk of injury to IAN based on Rood's Criteria in an intra oral periapical radiographic examination, between the age group of 18 to 40 years, were included in the study. Preoperatively the impacted third molars were evaluated clinically as well as radiographically. Pederson Difficulty Index and Winter's Classification of impacted tooth was recorded. Coronectomy was done at the cemento enamel junction leaving the roots 2-3mm below the alveolar crest and primary closure was done. Patients were evaluated periodically for two years at six months interval. Post operative pain, swelling, IAN injury or any other complications were observed and recorded. None of the patients had IAN injury and none required second surgical removal. There was no incidence of post-operative infection and none required second surgical intervention. However, two of our patients had failed coronectomy (10%) due to mobilization of roots intra operatively and the roots were removed. One patient developed profuse bleeding intra-operatively in the failed coronectomy case. One patient had temporary lingual nerve paresthesia. Coronectomy procedure is effective in controlling inferior alveolar nerve injury following third molar surgery, in radiographically evaluated high risk cases and it has very low incidence of complications.


Spontaneous hepatic rupture due to metastatic tumor of lung adenocarcinoma.
A 64-year-old man diagnosed as lung adenocarcinoma with hepatic tumor was admitted to our hospital. He carried the hepatitis B virus but was negative for PIVKA-II and alpha-fetoprotein, and hence we diagnosed a case of stage IV lung adenocarcinoma. We planned to administer systemic chemotherapy, but he experienced sudden-onset abdominal discomfort accompanied with decreased blood pressure. We diagnosed hemorrhagic ascites due to spontaneous rupture of the liver tumor. Emergency angiography and therapeutic embolization stabilized his clinical condition. Hemorrhagic ascites due to metastatic liver tumor is rare and the sudden onset of abdominal symptoms is an indicator of rupture.


(Pro)renin receptor as a therapeutic target for the treatment of cardiovascular diseases?
The discovery of the (pro)renin receptor [(P)RR] 15years ago stimulated ideas on prorenin being more than renin's inactive precursor. Indeed, binding of prorenin to the (P)RR induces a conformational change in the prorenin molecule, allowing it to display angiotensin-generating activity, and additionally results in intracellular signaling in an angiotensin-independent manner. However, the prorenin levels required to observe these angiotensin-dependent and -independent effects of the (P)RR are many orders above its in vivo concentrations, both under normal and pathological conditions. Given this requirement, the idea that the (P)RR has a function within the renin-angiotensin system (RAS) is now being abandoned. Instead, research is now focused on the (P)RR as an accessory protein of vacuolar H+-ATPase (V-ATPase), potentially determining its integrity. Acting as an adaptor between Frizzled co-receptor LRP6 and V-ATPase, the (P)RR appears to be indispensable for Wnt/β-catenin signaling, thus explaining why (P)RR deletion (unlike renin deletion) is lethal even when restricted to specific cells, such as cardiomyocytes, podocytes and smooth muscle cells. Furthermore, recent studies suggest that the (P)RR may play important roles in lipoprotein metabolism and overall energy metabolism. In this review, we summarize the controversial RAS-related effects of the (P)RR, and critically review the novel non-RAS-related functions of the (P)RR, ending with a discussion on the potential of targeting the (P)RR to treat cardiovascular diseases.


Regulation of fatty acid degradation in Escherichia coli: dominance studies with strains merodiploid in gene fadR.
Strains stably merodiploid in the 25-min region of the chromosome of Escherichia coli were constructed and used in dominance tests between various wild-type and mutant alleles of the fadR gene. Whereas the monoploid fadR+ and fadR strains were inducible and constitutive, respectively, for the enzymes involved in fatty acid degradation (fad), merodiploids with at least one fadR+ allele were inducible. This observation was true whether the fadR+ allele resided on the main chromosome or on the episome. These results show that fadR+ is trans dominant to fadR, and they are consistent with the proposal that the fadR gene product is a repressor protein. Complementation tests were also performed by constructing 24 merodiploids harboring fadR alleles on both the main chromosome and the episome. All of these fadR/fadR diploids were able to utilize the noninducing substrate decanoate as sole carbon source, suggesting that only one polypeptide is encoded by the fadR gene.


Multiresistant Salmonella Typhimurium DT104 infections of humans and domestic animals in the Pacific Northwest of the United States.
Salmonella Typhimurium definitive type 104 with chromosomally encoded resistance to five or more antimicrobial drugs (R-type ACSSuT+) has been reported increasingly frequently as the cause of human and animal salmonellosis since 1990. Among animal isolates from the northwestern United States (NWUS), R-type ACSSuT+ Typhimurium isolates increased through the early 1990s to comprise 73% of Typhimurium isolates by 1995, but subsequently decreased to comprise only 30% of isolates during 1998. NWUS S. Typhimurium R-type ACSSuT+ were consistently (99%) phage typed as DT104 or the closely related DTu302. S. Typhimurium isolates from cattle with primary salmonellosis, randomly selected from a national repository, from NWUS were more likely to exhibit R-type ACSSuT+ (19/24, 79%) compared to isolates from other quadrants (17/71, 24%; P < 0.01). Human patients infected with R-type ACSSuT+ resided in postal zip code polygons of above average cattle farm density (P < 0.05), while patients infected with other R-types showed no similar tendency. Furthermore, humans infected with R-type ACSSuT+ Typhimurium were more likely to report direct contact with livestock (P < 0.01) than humans infected with other R-types.


A pathway to personalization of integrated treatment: informatics and decision science in psychiatric rehabilitation.
Personalization of treatment is a current strategic goal for improving health care. Integrated treatment approaches such as psychiatric rehabilitation benefit from personalization because they involve matching diverse arrays of treatment options to individually unique profiles of need. The need for personalization is evident in the heterogeneity of people with severe mental illness and in the findings of experimental psychopathology. One pathway to personalization lies in analysis of the judgments and decision making of human experts and other participants as they respond to complex circumstances in pursuit of treatment and rehabilitation goals. Such analysis is aided by computer simulation of human decision making, which in turn informs development of computerized clinical decision support systems. This inspires a research program involving concurrent development of databases, domain ontology, and problem-solving algorithms, toward the goal of personalizing psychiatric rehabilitation through human collaboration with intelligent cyber systems. The immediate hurdle is to demonstrate that clinical decisions beyond diagnosis really do affect outcome. This can be done by supporting the hypothesis that a human treatment team with access to a reasonably comprehensive clinical database that tracks patient status and treatment response over time achieves better outcome than a treatment team without such access, in a controlled experimental trial. Provided the hypothesis can be supported, the near future will see prototype systems that can construct an integrated assessment, formulation, and rehabilitation plan from clinical assessment data and contextual information. This will lead to advanced systems that collaborate with human decision makers to personalize psychiatric rehabilitation and optimize outcome.


The Effect of the COVID-19 Pandemic on Childhood Immunizations: Ways to Strengthen Routine Vaccination.
Primary care providers play key roles in ensuring that children and the community receive vaccines on time. Sometimes, this role includes the task of reassuring parents who are vaccine hesitant that vaccines are safe and effective. Now, because of coronavirus disease 2019 (COVID-19), providers are presented with the additional challenge of maintaining and strengthening routine vaccination during a pandemic. As COVID-19 cases increased and states implemented stay-at-home orders, outpatient visits declined significantly. As a result, childhood immunization rates also declined. Increasing communication efforts regarding the importance of vaccination will be worthwhile, as the effect of the COVID-19 pandemic has highlighted the threat of an infectious disease and has increased awareness of the vaccine development process. Prior to 2020, many parents had not seen the devastating consequences of an infectious disease. The pandemic may change a parent's perspective, particularly as it relates to the influenza vaccine. Providers should continue to promote the importance of well-child and vaccination visits. [Pediatr Ann. 2020;49(12):e516-e522.].


Effect of chronic stress on expression and secretion of seminal vesicle proteins in adult rats.
Chronic stress (CS) is known to affect men's health especially fertility by reducing semen quality. Although the effects of CS on testicular function and sperm parameters are documented, changes of substances and secreting proteins in the seminal vesicle (SV) have never been reported. This study aimed to demonstrate the alterations of contents and expressions of proteins in seminal vesicle fluid (SVF) under CS. Fourteen adult rats were divided into control and CS groups (n = 7/each). Control rats were not exposed to stressor, while the CS animals were immobilised by restraint cage (4 hr/day) and followed by forced swimming (15 min/day) for consecutive 60 days. Biochemical substances and malondialdehyde (MDA) level in SVF were examined. Expressions of heat-shock protein 70 (Hsp70), caspases (Casp) 3 and 9, and tyrosine-phosphorylated (TyrPho) proteins were investigated in seminal vesicle tissue (SVT) and SVF. It was found that CS caused reductions of seminal epithelial height and secreted substance levels. Significantly, MDA levels in SVF and expressions of Hsp70, Casp and TyrPho proteins were increased in of CS animals. It was concluded that CS affected seminal secretion. Low quality of CS seminal plasma may associate with increase of MDA and expressions of secreted proteins.


Late vascular occlusion of central lines in pediatric malignancies.
To determine whether thrombi or vascular occlusion represent a late complication persisting several years after removal of central venous lines (CVLs) in children and adolescents treated for childhood cancer. Children whose treatment for malignancy included placement of a CVL that had been removed at least 2 months previously were studied during scheduled follow-up that included contrast-enhanced computed tomography. Spiral volume acquisition was used to obtain 3-mm images from the chest apices through the right hilum, and three-dimensional reconstruction of angiograms was performed. Thrombosis/occlusion was defined as narrowing, obstruction, or filling defect of the deep venous system, with or without the formation of collateral veins. Charts were reviewed to document patient characteristics, previous CVL complications, administration of hyperalimentation, use of urokinase, and family history of venous thrombosis. Twenty-three patients treated for solid tumors and 2 treated for B-cell acute lymphocytic leukemia or lymphoma were studied. Lines had been in place from 0.2 to 36 months (median, 7.4) and were removed at 2.3 to 121.8 months (median, 32.5) before study. Nine patients received hyperalimentation for periods ranging from 2 to 38 weeks (median, 12). Four patients had required urokinase instillations, and one developed superior vena cava syndrome; 4 had a CVL-related infection (two superficial and two Candida line infections). Occlusion was seen on computed tomography angiograms in 3 of the 25 patients (12%; 95% confidence interval: 4.5-31%). One of the patients with occlusion had superior vena cava syndrome; none had a family history of thrombosis, use of a double lumen CVL, or multiple instillations of urokinase. Persistent asymptomatic vascular occlusion does occur as a late complication of CVL placement for treatment of childhood malignancies, although the frequency appears low among patients treated primarily for solid tumors. Prospective studies of large numbers of patients with a broader spectrum of diagnoses are necessary to define the incidence of and risk factors for this complication and to assess the need for prevention with anticoagulation or other therapy. Pediatricians caring for patients with a history of cancer and CVLs should be aware that these patients may have persistent vascular occlusion that could predispose them to recurrent thrombosis or postphlebitic syndrome.


Measurement setup to simultaneously explore the location and energy of trapped charges in thin polymer films.
This research proposes a unique system that combines charge density measurements by the laser intensity modulation method with optically excited current acquisitions using the photo-stimulated discharge technique (PSD). The purpose of this setup is to investigate the relationship between space charge properties (such as density, spatial depth, and time evolution) and the photocurrent-associated energies in order to gain new insights into the trap population and detrapping mechanisms in thin polymer films. This paper presents a description of the technical principles of both methods as well as the whole combined system. The results on a 12 μm-thick polyethylene naphthalate film show pyroelectric currents, which after processing indicate the formation of homocharges whose magnitude and depth decrease after light irradiation. The PSD currents allow the identification of two major energy bands at 3.4 and 5.9 eV (360 and 207 nm, respectively), possibly related to charge detrapping. In addition, current transients during constant wavelength irradiation show that incident photons can interact differently with trapped charges depending on the applied field.


[Splanchnic hemodynamics in acute cholecystitis and mechanical jaundice].
The authors analyze results of the investigation of hemodynamics in the abdominal cavity vessels by the method of duplex scanning in 30 patients with acute cholecystitis, in 30 patients with mechanical jaundice and cholelithiasis and in 30 healthy subjects. It was found that these patients had less blood flow along the upper mesenterial artery and the portal vein. As a compensation the blood flow increased along the hepatic artery.


Mn(II) oxidation in Pseudomonas putida GB-1 is influenced by flagella synthesis and surface substrate.
Bacterially mediated manganese(II) oxidation greatly affects the biogeochemical cycling of Mn and other elements. One species of bacteria that are capable of Mn(II) oxidation is the gamma-proteobacterium Pseudomonas putida GB-1. In this organism, Mn(II) oxidation begins in stationary phase on the outer surface of the cell, forming a layer of insoluble Mn(III,IV) oxides. A random transposon mutagenesis screen isolated 12 mutant strains of P. putida GB-1 that exhibited increased Mn(II) oxidation on solid media relative to wild type. In 8 out of the 12 strains, the transposon had inserted into a putative flagellar gene. Those 8 strains each had motility defects, thus the disrupted genes are part of the P. putida GB-1 flagellar regulon. The flagellar genes identified include putative structural components (FliC, FliD, FlgE, and FlgL) and regulatory proteins (FlgM and FleN). Deletion of either the FleN gene (fleN) or the overlapping gene fliA resulted in increased Mn(II) oxidation, while in-frame deletion of fliF, which encodes an essential component of the basal body, did not. In liquid media, the flagellar mutants exhibited delayed Mn(II) oxidation relative to wild type. These results suggest that bacterial Mn(II) oxidation is regulated in part by flagellar-mediated responses to the surface substrate.


Enhancement of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA amplification in a Simian virus 40-transformed Chinese hamster cell line by 3-aminobenzamide.
A Simian virus 40-transformed Chinese hamster cell line (CO 60) amplifies integrated viral DNA sequences as a response to treatment with a variety of carcinogens. To study a possible involvement of poly(ADP-ribose) synthesis, DNA amplification was induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), an alkylating carcinogen that strongly stimulates poly(ADP-ribose) synthesis. In the presence of 3-aminobenzamide (3AB) (2 mM), a competitive inhibitor of poly(ADP-ribose) polymerase, MNNG-induced amplification was increased two to six times the level induced by MNNG alone. Concomitantly, 3AB reduced cellular poly(ADP-ribose) levels and increased MNNG-induced cytotoxicity, as expected. The effect of 3AB on MNNG-induced amplification depended both on the concentration of 3AB and the duration of its presence after MNNG treatment. By contrast, 3-aminobenzoic acid, a noninhibitory structural analogue of 3AB, had no influence on amplification induced by MNNG. These data strongly suggest an involvement of poly(ADP-ribose) in the process of DNA amplification, as it is shown that inhibition of carcinogen-stimulated poly(ADP-ribose) synthesis by 3AB is correlated with an enhancement of inducible DNA amplification in this cell line.


Microgel capsules tailored by droplet-based microfluidics.
Microgel capsules are micrometer-sized particles that consist of a cross-linked, solvent-swollen polymer network complexed with additives. These particles have various applications, such as drug delivery, catalysis, and analytics. To optimize the performance of microgel capsules, it is crucial to control their size, shape, and content of encapsulated additives with high precision. There are two classes of microgel-capsule structures. One class comprises bulk microcapsules that consist of a polymer network spanning the entire particle and entrapping the additive within its meshes. The other class comprises core-shell structures; in this case, the microgel polymer network just forms the shell of the particles, whereas their interior is hollow and hosts the encapsulated payload. Both types of structures can be produced with exquisite control by droplet-based microfluidic templating followed by subsequent droplet gelation. This article highlights some early and recent achievements in the use of this technique to tailor soft microgel capsules; it also discusses applications of these particles. A special focus is on the encapsulation of living cells, which are very sensitive and complex but also very useful additives for immobilization within microgel particles.


Human-Genetic Ancestry Inference and False Positives in Forensic Familial Searching.
In forensic familial search methods, a query DNA profile is tested against a database to determine if the query profile represents a close relative of a database entrant. One challenge for familial search is that the calculations may require specification of allele frequencies for the unknown population from which the query profile has originated. The choice of allele frequencies affects the rate at which non-relatives are erroneously classified as relatives, and allele-frequency misspecification can substantially inflate false positive rates compared to use of allele frequencies drawn from the same population as the query profile. Here, we use ancestry inference on the query profile to circumvent the high false positive rates that result from highly misspecified allele frequencies. In particular, we perform ancestry inference on the query profile and make use of allele frequencies based on its inferred genetic ancestry. In a test for sibling matches on profiles that represent unrelated individuals, we demonstrate that false positive rates for familial search with use of ancestry inference to specify the allele frequencies are similar to those seen when allele frequencies align with the population of origin of a profile. Because ancestry inference is possible to perform on query profiles, the extreme allele-frequency misspecifications that produce the highest false positive rates can be avoided. We discuss the implications of the results in the context of concerns about the forensic use of familial searching.


Single center experience with total body irradiation and melphalan (TBI-MEL) myeloablative conditioning regimen for allogeneic stem cell transplantation (SCT) in patients with refractory hematologic malignancies.
We retrospectively evaluated the tolerability and efficacy of fractionated total body irradiation (TBI) (1,200 cGy) and melphalan (MEL) (100-110 mg/m(2)) myeloablative conditioning in 48 patients with nonremission AML (n = 14), ALL (n = 10), NHL (n = 18), and other refractory hematologic malignancies (n = 6) who received allogeneic stem cell transplantation (SCT) between 2002 and 2011. Median age was 48 years (22 to 68); 14 out of 26 leukemia patients (54 %) had circulating blasts at transplant, 20 (50 %) evaluable patients had poor-risk cytogenetics, 12 (25 %) had prior SCT, and 10 (21 %) received stem cells from a mismatch donor. All patients received tacrolimus with or without methotrexate for GVHD prophylaxis. At the time of analysis, 13 patients (27 %) were alive and disease free. Engraftment was complete in all patients. The median time to ANC recovery (>500) was 12 days (range, 6-28). The most common grade III and IV toxicities were mucositis and infections. Eighteen patients (43 %) developed grade II-IV acute GVHD, and eight (26 %) had extensive chronic GVHD. Of 44 evaluable patients for response, 28 (64 %) achieved a complete remission (CR), and seven (15 %) had a partial remission after the transplant. With a median follow-up of 30 months (4 to 124 months) for surviving patients, the cumulative incidence of relapse was 45 % at 1 year, and the probability of overall survival (OS) at 5 years was 22.5 %. Multivariate analysis showed that platelet count (<80,000/mL) and lactic dehydrogenase (>500 IU/L) at SCT were associated with relapse. Age less than 53 years and CR after SCT were associated with better OS. Our data suggest that TBI-MEL can result in CR in two thirds, durable remission in one third, and 5-year survival in about one quarter of patients with nonremission hematologic malignancies. Further studies with TBI-MEL in standard risk transplant patients are warranted.


Novelty enhances visual perception.
The effects of novelty on low-level visual perception were investigated in two experiments using a two-alternative forced-choice tilt detection task. A target, consisting of a Gabor patch, was preceded by a cue that was either a novel or a familiar fractal image. Participants had to indicate whether the Gabor stimulus was vertically oriented or slightly tilted. In the first experiment tilt angle was manipulated; in the second contrast of the Gabor patch was varied. In the first, we found that sensitivity was enhanced after a novel compared to a familiar cue, and in the second we found sensitivity to be enhanced for novel cues in later experimental blocks when participants became more and more familiarized with the familiar cue. These effects were not caused by a shift in the response criterion. This shows for the first time that novel stimuli affect low-level characteristics of perception. We suggest that novelty can elicit a transient attentional response, thereby enhancing perception.


Two faces of vitamin E in the lung.
Asthma and allergic lung disease occur as complex environmental and genetic interactions. Clinical studies of asthma indicate a number of protective dietary factors, such as vitamin E, on asthma risk. However, these studies have had seemingly conflicting outcomes. In this perspective, we discuss opposing regulatory effects of tocopherol isoforms of vitamin E, mechanisms for tocopherol isoform regulation of allergic lung inflammation, association of vitamin E isoforms with outcomes in clinical studies, and how the variation in global prevalence of asthma may be explained, at least in part, by vitamin E isoforms.