OpenCASA: A new open-source and scalable tool for sperm quality analysis.
In the field of assisted reproductive techniques (ART), computer-assisted sperm analysis (CASA) systems have proved their utility and potential for assessing sperm quality, improving the prediction of the fertility potential of a seminal dose. Although most laboratories and scientific centers use commercial systems, in the recent years certain free and open-source alternatives have emerged that can reduce the costs that research groups have to face. However, these open-source alternatives cannot analyze sperm kinetic responses to different stimuli, such as chemotaxis, thermotaxis or rheotaxis. In addition, the programs released to date have not usually been designed to encourage the scalability and the continuity of software development. We have developed an open-source CASA software, called OpenCASA, which allows users to study three classical sperm quality parameters: motility, morphometry and membrane integrity (viability) and offers the possibility of analyzing the guided movement response of spermatozoa to different stimuli (useful for chemotaxis, thermotaxis or rheotaxis studies) or different motile cells such as bacteria, using a single software. This software has been released in a Version Control System at Github. This platform will allow researchers not only to download the software but also to be involved in and contribute to further developments. Additionally, a Google group has been created to allow the research community to interact and discuss OpenCASA. For validation of the OpenCASA software, we analysed different simulated sperm populations (for chemotaxis module) and evaluated 36 ejaculates obtained from 12 fertile rams using other sperm analysis systems (for motility, membrane integrity and morphology modules). The results were compared with those obtained by Open-CASA using the Pearson's correlation and Bland-Altman tests, obtaining a high level of correlation in all parameters and a good agreement between the different used methods and the OpenCASA. With this work, we propose an open-source project oriented to the development of a new software application for sperm quality analysis. This proposed software will use a minimally centralized infrastructure to allow the continued development of its modules by the research community.


Congenital syphilis: trends in mortality and morbidity in the United States, 1999 through 2013.
Congenital syphilis (CS) results when an infected pregnant mother transmits syphilis to her unborn child prior to or at delivery. The severity of infection can range from a delivery at term without signs of infection to stillbirth or death after delivery. We sought to describe CS morbidity and mortality during 1999 through 2013. National CS case data reported to Centers for Disease Control and Prevention during 1999 through 2013 were analyzed. Cases were classified as dead (stillbirths and deaths up to 12 months after delivery), morbid (cases with strong [physical, radiographic, and/or nonserologic laboratory] evidence of CS), and nonmorbid (cases with a normal physical examination reported, without strong evidence of infection). Annual rates of these cases were calculated. Cases were compared using selected maternal and infant criteria. During 1999 through 2013, 6383 cases of CS were reported: 6.5% dead, 33.6% morbid, 53.9% nonmorbid, and 5.9% unknown morbidity; 81.8% of dead cases were stillbirths. Rates of dead, morbid, and nonmorbid cases all decreased over this time period, but the overall proportions that were dead or morbid cases did not significantly change. The overall case fatality ratio during 1999 through 2013 was 6.5%. Among cases of CS, maternal race/ethnicity was not associated with increased morbidity or death, although most cases (83%) occurred among black or Hispanic mothers. No or inadequate treatment for maternal syphilis, <10 prenatal visits, and maternal nontreponemal titer ≥1:8 increased the likelihood of a dead case; risk of a dead case increased with maternal nontreponemal titer (χ(2) for trend P < .001). Infants with CS born alive at <28 weeks' gestation (relative risk, 107.4; P < .001) or born weighing <1500 g (relative risk, 43.9; P < .001) were at greatly increased risk of death. CS remains an important preventable cause of perinatal morbidity and mortality, with comparable case fatality ratios during 1999 through 2013 (6.5%) and 1992 through 1998 (6.4%). Detection and treatment of syphilis early during pregnancy remain crucial to reducing CS morbidity and mortality.


Maternal practice and attitudes toward breastfeeding.
An age-graded sample of 411 New York City mothers is assessed to evaluate the direction and amount of change in incidence of breastfeeding. Social and demographic factors are considered and are found to have little trend effect. This limited sample fails to indicate any increase in breastfeeding in the 1970s, as compared to earlier decades. However, maternal attitudes are more positive toward breastfeeding than toward bottlefeeding; and some ethnic, economic, and sociocultural factors do affect decision to breastfeed.


ONTOGENETIC STUDIES OF FLORETS IN POA (GRAMINEAE): ALLOMETRY AND HETEROCHRONY.
Species distinctions in grasses are based on differences in sizes of parts, particularly floral parts, and on differences in distribution of hairs; both sets of characters are apparently subject to repeated parallel evolution. Developmental studies of florets in the genus Poa reveal that there are only two developmental trajectories for lemma length relative to anther length (a measure of time to maturity): either lemma growth is continuous with anther growth, or the lemma reaches its adult size well before the anthers do. The tuft of long, cobweb-like hairs on the callus of many species forms early in lemma development, as might be expected for a character that marks a large group of species. The five possible patterns of hair distribution on the lemma itself represent stages of a single developmental pathway; differences between species presumably reflect differences in timing of onset or truncation of the developmental program (heterochrony).


α-Radical Phosphines: Synthesis, Structure, and Reactivity.
A series of phosphines featuring a persistent radical were synthesized in two steps by condensation of dialkyl-/diarylchlorophosphines with stable cyclic (alkyl)(amino)carbenes (cAACs) followed by one-electron reduction of the corresponding cationic intermediates. Structural, spectroscopic, and computational data indicate that the spin density in these phosphines is mainly localized on the original carbene carbon from the cAAC fragment; thus, it remains in the α-position with respect to the central phosphorus atom. The potential of these α-radical phosphines to serve as spin-labeled ligands is demonstrated through the preparation of several AuI derivatives, which were also structurally characterized by single-crystal X-ray diffraction.


Anatomy of news consumption on Facebook.
The advent of social media and microblogging platforms has radically changed the way we consume information and form opinions. In this paper, we explore the anatomy of the information space on Facebook by characterizing on a global scale the news consumption patterns of 376 million users over a time span of 6 y (January 2010 to December 2015). We find that users tend to focus on a limited set of pages, producing a sharp community structure among news outlets. We also find that the preferences of users and news providers differ. By tracking how Facebook pages "like" each other and examining their geolocation, we find that news providers are more geographically confined than users. We devise a simple model of selective exposure that reproduces the observed connectivity patterns.


Caffeic Acid-Zinc Basic Salt/Chitosan Nanohybrid Possesses Controlled Release Properties and Exhibits In Vivo Anti-Inflammatory Activities.
Caffeic acid (CA) exhibits a myriad of biological activities including cardioprotective action, antioxidant, antitumor, anti-inflammatory, and antimicrobial properties. On the other hand, CA presents low water solubility and poor bioavailability, which have limited its use for therapeutic applications. The objective of this study was to develop a nanohybrid of zinc basic salts (ZBS) and chitosan (Ch) containing CA (ZBS-CA/Ch) and evaluate its anti-edematogenic and antioxidant activity in dextran and carrageenan-induced paw edema model. The samples were obtained by coprecipitation method and characterized by X-ray diffraction, Fourier transform infrared (FT-IR), scanning electron microscope (SEM) and UV-visible spectroscopy. The release of caffeate anions from ZBS-CA and ZBS-CA/Ch is pH-dependent and is explained by a pseudo-second order kinetics model, with a linear correlation coefficient of R2 ≥ 0.99 at pH 4.8 and 7.4. The in vivo pharmacological assays showed excellent anti-edematogenic and antioxidant action of the ZBS-CA/Ch nanoparticle with slowly releases of caffeate anions in the tissue, leading to a prolongation of CA-induced anti-edematogenic and anti-inflammatory activities, as well as improving its inhibition or sequestration antioxidant action toward reactive species. Overall, this study highlighted the importance of ZBS-CA/Ch as an optimal drug carrier.


Solid state characterization of spray-dried powders of recombinant human deoxyribonuclease (rhDNase).
rhDNase is a recombinant human protein approved as an aqueous solution for human use by inhalation. To study the feasibility of preparation of dry powders for inhalation, spray-dried powders of pure rhDNase and co-spray-dried mixtures of rhDNase with an excipient approved for inhalation products, lactose, were prepared. Both types of powders were initially amorphous. The lactose, however, was found to crystallize after exposure to a humid environment. The crystals in the powder were identified as the alpha-monohydrate polymorph of lactose by hot-stage optical and scanning electron microscopy, differential scanning calorimetry and thermogravimetry, FTIR spectroscopy, and X-ray powder diffraction. Moisture sorption isotherms indicated that crystallization occurred at high relative humidities (70-85%), depending on the temperature of the environment (5-40 degreesC). The practical implications for the manufacturing and storage of protein powders for inhalation are discussed.


Epilepsy and clefting disorders: lack of evidence of a familial association.
Offspring of women with epilepsy who are taking anticonvulsant medications have an increased incidence of clefting abnormalities. However, there is disagreement as to the extent to which this increase is due to a teratogenic effect of anticonvulsant medications or to a familial association of epilepsy and clefting. Some investigators suggested, but were unable to demonstrate adequately, that clefting and epilepsy associate together in families. In order to address this issue, individuals with a clefting disorder and their relatives were ascertained to determine whether clefting and epilepsy aggregate together in families. The incidence of epilepsy was not increased over that in the general population with a standardized morbidity ratio (SMR) for epilepsy of 0.9 (95% CI 0.2-2.6) for first-degree relatives (excluding parents) and 0.0 for second-degree relatives. The SMRs ranged from 0.7-2.2 for the individual relative types (parents 1.5, sibs 0.7, children 2.2) and were also not increased. These results do not support suggestions that clefting and epilepsy segregate together in families.


How much POCUS for Czech internists?
The introduction of point-of-care ultrasonography into practice in internal medicine and subsequently into the educational program for our specialty brought the need to define a curriculum for training in this method. We solve the question of "what to teach" - i.e. what core ultrasound competencies internists should learn for their practice. It is also necessary to define the procedure "how to teach" - the form and content of the education program, what the basic course should contain, and above all how the subsequent training should take place in practice. The third major problem to be solved is "who should teach", i.e. the definition of the requirements for trainers who will lead the training.


The antiapoptotic effect of galectin-3 in human endometrial cells under the regulation of estrogen and progesterone.
Galectin-3 (Gal-3), a ubiquitously expressed gene involved in many cellular processes, has been recently recognized as a factor related to endometrial receptivity. However, the precise biological function of Gal-3 in the endometrium and its regulation is still unclear. In this study, we detected the antiapoptotic role of Gal-3 in endometrial cells and the expression of Gal-3 regulated by estrogen and progesterone. We found that expression of Gal-3 increased when exposed to the apoptosis inducer staurosporine. Gal-3-silenced endometrial cells were more sensitive to the apoptosis inducer. Estradiol (E2) and progesterone (P4) up-regulated Gal-3 expression, which in turn decreased the apoptotic rate of endometrial cells. Our results strongly suggested that hormonal activation of Gal-3 by E2 and P4 is involved in inhibiting endometrial cell apoptosis, playing key roles in embryo implantation.


The Distal Stump of the Intramuscular Motor Branch of the Obturator Nerve Is Useful for the Reconstruction of Long-Standing Facial Paralysis Using a Double-Powered Free Gracilis Muscle Flap Transfer.
Double innervation of the transferred muscle with the contralateral facial nerve and the ipsilateral masseteric nerve has recently been reported by some authors. The aim of this study was to assess the utility of our procedure of double innervation of free gracilis muscle for reconstruction of long-standing facial palsy. In our department, 6 cases of long-standing facial paralysis (4 cases of complete palsy and 2 of incomplete palsy) were reconstructed using a free gracilis muscle double innervated with the masseteric and contralateral facial nerves. The patient age ranged from 37 to 79 years (average 56.7 ± 15.7). In our procedure, the intramuscular motor branch of the transferred muscle was identified and sutured to the ipsilateral masseteric nerve in an end-to-end fashion, and the obturator nerve of the transferred muscle was sutured to the cross-facial nerve graft, which was coapted with the contralateral facial nerve by end-to-end suturing. All patients were followed up for >18 months and recovered their smiling function. The voluntary movement of the transferred muscle with teeth clenching was observed at approximately 4.7 months, and this movement combined with contralateral mouth angle elevation was observed at approximately 9.5 months after the surgery. Our experience suggests that the distal stump of the intramuscular motor branch of the obturator nerve may be useful for facial reanimation via double-powered free gracilis muscle flap transfer.


Vascular endothelial growth factor-A mediates ultraviolet B-induced impairment of lymphatic vessel function.
UVB irradiation of the skin induces erythema, epidermal hyperplasia, vascular hyperpermeability, and edema formation. Previous studies have revealed that the cutaneous blood vasculature plays a critical role in the mediation of photodamage. In contrast, the role of lymphatic vessels, which play an essential role in the maintenance of tissue fluid balance, in the response to UVB irradiation has remained unknown. We report here that both acute and chronic UVB irradiation of murine skin results in prominent enlargement of lymphatic vessels. Surprisingly, these enlarged lymphatic vessels were functionally impaired and hyperpermeable, as detected by intravital lymphangiography. The expression levels of vascular endothelial growth factor (VEGF)-A but not of the known lymphangiogenesis factors VEGF-C or VEGF-D, were enhanced in UVB-irradiated epidermis. Targeted overexpression of VEGF-A in the epidermis of transgenic mice led to increased enlargement and leakage of lymphatic vessels after acute UVB irradiation, whereas systemic blockade of VEGF-A signaling largely prevented lymphatic vessel abnormalities and photodamage induced by UVB. Together, these findings identify lymphatic vessels as novel targets for UVB-induced cutaneous photodamage and suggest that VEGF-A mediates impairment of lymphatic vessel function, thereby contributing to the adverse effects of UVB irradiation on the skin.


Effects of wheat or corn distillers dried grains with solubles on feedlot performance, fecal shedding, and persistence of Escherichia coli O157:H7.
Distillers dried grains with solubles (DDGS) are a coproduct of the ethanol industry and are often used as a replacement for grain in livestock production. Feeding corn DDGS to cattle has been linked to increased fecal shedding of Escherichia coli O157:H7, although in Canada, DDGS are often produced from wheat. This study assessed the effects of including 22.5% wheat or corn DDGS (DM basis) into barley-based diets on performance, carcass characteristics, animal health, and fecal E. coli O157:H7 shedding of commercial feedlot cattle. Cattle (n = 6,817) were randomly allocated to 10 pens per treatment group: WDDGS (diets including 22.5% wheat DDGS), CDDGS (diets including 22.5% corn DDGS), or CTRL (barley substituted for DDGS). Freshly voided fecal pats (n = 588) were collected and pooled monthly for fecal pH measurement and screened for naturally occurring E. coli O157:H7 by immunomagnetic separation (IMS) and direct plating (DP). Hide swabs (n = 367) were collected from randomly selected cattle from each pen before slaughter. Pen-floor fecal samples (n = 18) were collected from treatment groups at entry to the feedlot (<14 d on the finishing diet) and after adapting to the finishing diet for ≥ 14 d, inoculated (10(9) cfu of a 5 strain naldixic acid-resistant E. coli O157:H7 mixture), incubated (20°C) and evaluated weekly (IMS and DP) to assess fecal E. coli O157:H7 persistence. The WDDGS group had 3.0% poorer ADG (P = 0.007), 5.3% poorer G:F (P < 0.001), and a decreased proportion of Canada Quality Grade AAA carcasses (P = 0.022) compared with CTRL cattle. The CDDGS group had a similar ADG (P = 0.06), a decreased proportion of Canada Yield Grade (YG) 1 (P < 0.001), and greater proportions of Canada YG 2 (P = 0.003) and YG 3 (P < 0.001) carcasses compared with the CTRL group. There were no differences among groups in any of the animal health parameters assessed. Inclusion of DDGS in cattle finishing diets had no effect on fecal shedding (P = 0.650) or persistence (P = 0.953) of E. coli O157:H7. However, feces from cattle on starter diets <14 d had longer persistence of E. coli O157:H7 (week) than cattle on finishing diets ≥ 14 d (P < 0.003). Inclusion of DDGS in feedlot diets depends on commodity pricing relative to that of barley and for WDDGS must also include the risk of feedlot performance and carcass grading disadvantages. Feeding cattle barley based-diets with 22.5% corn or wheat DDGS did not affect fecal shedding of E. coli O157:H7.


Destruction of the germinal disc region of an immature preovulatory follicle suppresses follicular maturation and ovulation.
Previous work has suggested that the germinal disc and the overlying layer of granulosa cells located near the germinal disc, collectively referred to as the germinal disc region (GDR), is a primary "growth center," regulating granulosa cell proliferation within follicles in the rapid growth phase. This study was designed to determine whether the presence of a viable GDR is required for the completion of follicular maturation and ovulation of the avian follicle. Twelve or 24 h before the expected time of ovulation, the GDR of the largest preovulatory follicle (F1) was destroyed by applying solid CO2 (2 x 2 mm) for 20 sec. A region of the follicle wall equal in size to but opposite the GDR was destroyed in control birds. Blood samples were taken 24 h and 6 h before and at the time of ovulation. The effects of GDR destruction on plasma progesterone (P4), LH, and ovulation were determined. Destruction of the F1 GDR 24 h before ovulation resulted in an absence of the preovulatory rise in plasma P4, attenuation of the LH surge, blocked ovulation, and atresia of the F1 follicle. Controls displayed typical preovulatory profiles of plasma P4 and LH and ovulated at the expected time. In contrast to the data collected after destruction of the GDR 24 h before ovulation, destruction of the GDR 12 h before ovulation did not disrupt ovulation. Furthermore, destruction of the GDR 24 h before ovulation had no effect on basal or LH-stimulated P4 production by granulosa cell cultures prepared 12 h after destruction of the GDR.(ABSTRACT TRUNCATED AT 250 WORDS)


Suicide and the MMPI: a cross-validation of predictors.
Compared a suicide-completion group to a psychiatric control sample on the 13 traditional MMPI scales, three experimental item pools, and eight profile patterns earlier described as indicative of suicidal tendencies (N = 84). Also compared the frequencies with which the groups endorsed each of the 566 MMPI items. The number of resulting significant differences was less than chance. The results argue against the use of the MMPI to predict suicide at this time.


The dystrophic hamster: an animal model of alveolar hypoventilation.
The BIO 14.6 dystrophic hamster (DH) is a genetically determined animal model of alveolar hypoventilation (AH) that exhibits a ventilatory control pattern of compensation and then decompensation similar to that in progressive forms of muscular dystrophy and nonprogressive congenital myopathies in humans. Possible causes of AH in the DH include respiratory muscle weakness, ventilation-to-perfusion inequalities, and an inadequate drive to breathe. Histochemical and contractile abnormalities of the diaphragm, reduced lung surface area available for gas exchange, abnormal pulmonary microvascular reactivity to hypoxia, altered levels of neurochemicals, and abnormal cellular regulation of calcium are among the specific factors that may contribute to the development of AH. The potential role of hypothyroidism in the development of AH is reviewed because many hypothyroid patients exhibit AH and other ventilatory dysfunctions, hypothyroidism is present in human patients and animals with muscular dystrophy, and thyroid status is known to influence lung architecture, myocyte function, and neural activity. Additional studies linking neurohormonal signals, transcellular signal processing, and control of ventilation in the DH may help us understand the etiology of AH in human disease.


Gastrointestinal neuroendocrine neoplasms (GI-NENs): hot topics in morphological, functional, and prognostic imaging.
Neuroendocrine neoplasms (NENs) are heterogeneous tumours with a common phenotype descended from the diffuse endocrine system. NENs are found nearly anywhere in the body but the most frequent location is the gastrointestinal tract. Gastrointestinal neuroendocrine neoplasms (GI-NENs) are rather uncommon, representing around 2% of all gastrointestinal tumours and 20-30% of all primary neoplasms of the small bowel. GI-NENs have various clinical manifestations due to the different substances they can produce; some of these tumours appear to be associated with familial syndromes, such as multiple endocrine neoplasm and neurofibromatosis type 1. The current WHO classification (2019) divides NENs into three major categories: well-differentiated NENs, poorly differentiated NENs, and mixed neuroendocrine-non-neuroendocrine neoplasms. The diagnosis, localization, and staging of GI-NENs include morphology and functional imaging, above all contrast-enhanced computed tomography (CECT), and in the field of nuclear medicine imaging, a key role is played by 68Ga-labelled-somatostatin analogues (68Ga-DOTA-peptides) positron emission tomography/computed tomography (PET/TC). In this review of recent literature, we described the objectives of morphological/functional imaging and potential future possibilities of prognostic imaging in the assessment of GI-NENs.


Automatic feature-based grouping during multiple object tracking.
Contour interpolation automatically binds targets with distractors to impair multiple object tracking (Keane, Mettler, Tsoi, & Kellman, 2011). Is interpolation special in this regard or can other features produce the same effect? To address this question, we examined the influence of eight features on tracking: color, contrast polarity, orientation, size, shape, depth, interpolation, and a combination (shape, color, size). In each case, subjects tracked 4 of 8 objects that began as undifferentiated shapes, changed features as motion began (to enable grouping), and returned to their undifferentiated states before halting. We found that intertarget grouping improved performance for all feature types except orientation and interpolation (Experiment 1 and Experiment 2). Most importantly, target-distractor grouping impaired performance for color, size, shape, combination, and interpolation. The impairments were, at times, large (>15% decrement in accuracy) and occurred relative to a homogeneous condition in which all objects had the same features at each moment of a trial (Experiment 2), and relative to a "diversity" condition in which targets and distractors had different features at each moment (Experiment 3). We conclude that feature-based grouping occurs for a variety of features besides interpolation, even when irrelevant to task instructions and contrary to the task demands, suggesting that interpolation is not unique in promoting automatic grouping in tracking tasks. Our results also imply that various kinds of features are encoded automatically and in parallel during tracking.


A comprehensive investigation of cardiac arrest before and after arrival of emergency medical services.
Many of the factors that affect survival from out-of-hospital cardiac arrest are not relevant in patients who arrest after arrival of emergency medical services (EMS). Because all arrests that occur after arrival of EMS are witnessed and care is immediate, one might expect survival to be very high. Several studies have described communities' experiences of arrest after arrival but few have compared survival rates stratified by rhythm and witness status. The purpose of this paper was to describe the characteristics of patients who arrested after arrival of EMS and to compare survival in this population to those who had witnessed and unwitnessed arrests before EMS arrival. We conducted a retrospective cohort study in King County, WA, USA. Descriptive statistics were calculated in patients whose arrests were not witnessed, in patients whose arrests were witnessed by citizens, and in those whose arrests were witnessed by EMS personnel. The majority of bystander- and EMS-witnessed arrests were initially in ventricular fibrillation (VF), but EMS-witnessed arrests were more likely to initially have been in pulseless electrical activity (PEA) than bystander-witnessed events. Patients whose arrests were witnessed by EMS had the greatest likelihood of survival compared to patients whose arrests were not witnessed or were witnessed by bystanders. Patients whose arrests were witnessed by EMS and were initially in VF had the highest rates of survival (59%). Patients whose arrests were witnessed by EMS were more likely to have survived their cardiac arrests than those who arrested before EMS arrived. We suggest that survival rates from VF arrests that occur after EMS arrival should be widely reported in order to measure overall EMS performance since many factors such as response times, bystander actions, and witness status are equalized in this subset of patients.


Incidence of Preventable Nonfatal Craniofacial Injuries and Implications for Facial Transplantation.
The number of patients who may benefit from evaluation for face transplantation in the United States (US) remains largely unknown. The goal of our study was to better delineate the pool of patients who might benefit from face transplant evaluation based on the characteristics and mechanisms of injury of previously reported face transplant recipients. The authors utilized data from the National Electronic Injury Surveillance System-All Injury Program in this study. The US Census Bureau data were used for population estimates. Inclusion and exclusion criteria were determined based on the characteristics of face transplant recipients to date, and the mechanisms of injury they sustained ultimately necessitating face transplantation. Statistical significance was reached if P <0.05. The estimated annual incidence of preventable craniofacial injuries from firearms (44,266-58,299; 31.7% increase), burns (5712-19,433; 240.2% increase), and animal attacks (5355-14,666; 173.9% increase) increased from 2005 to 2014, whereas the estimated annual incidence of craniofacial injuries from machinery (3927-2933; 25.3% decrease) decreased between 2005 and 2014. The authors estimate the annual incidence rate to fall between 32.1 per 100,000 and 58.1 per 100,000 among individuals aged 20 to 64 in the US. In this study, the authors estimate the annual incidence rate of individuals aged 20 to 64 in the US who may benefit from face transplant evaluation and believe that this quantification has the potential to initiate actionable discussions regarding geographical and financial factors affecting access to care in this patient population.


Lineage specification revealed by single-cell gene expression analysis in porcine preimplantation embryos.
After zygotic genome activation and lineage specification, zygotes develop into late blastocysts comprising three distinct cell types. The molecular mechanisms underlying this progress are largely unknown in pigs. Here, we intended to analyze an extensive set of regulators at the single-cell level to define the events involved in the development of the porcine blastocysts. Using a quantitative microfluidics approach in single cells, we detected mRNA levels of 96 genes known to function in early embryonic development and maintenance of stem cell pluripotency simultaneously in 480 individual cells derived from porcine preimplantation embryos. The developmental transitions can be distinguished based on distinctive gene expression profiles, and we identified paired box 6 (PAX6) and aquaporin 3 (AQP3) expressed in early and late developmental stages, respectively. Two lineages can be segregated in porcine early and late blastocysts by the expression patterns of lineage-specific genes such as DAB2, clathrin adaptor protein (DAB2) for trophectoderm (TE), platelet derived growth factor receptor alpha (PDGFRA), Nanog homeobox (NANOG), fibronectin 1 (FN1), hepatocyte nuclear factor 4 alpha (HNF4A), goosecoid homeobox (GSC), nuclear receptor subfamily 5 group A member 2 (NR5A2), and lysine acetyltransferase 6A (KAT6A; previously known as MYST3) for inner cell mass (ICM). However, the epiblast and primitive endoderm cannot be identified in late blastocysts, and those TE or ICM lineage-specific genes were low expressed in blastomeres from the morula. Our results shed light on early cell fate determination in porcine preimplantation embryos and offer theoretical support for deriving porcine embryonic stem cells.


An Original Suprapannicular Incision Technique for Cesarean Delivery in the Morbidly Obese Parturient.
Obese patients are at an increased risk of cesarean delivery and its associated wound complications. We present an alternative incision technique for obese parturients that avoids making a Pfannenstiel incision under the panniculus while still providing access to the lower uterine segment. For our technique, an assistant uses Allis clamps to exert caudal traction on the patient's panniculus and a transverse skin incision is made under the umbilicus, 3 cm below the line joining the anterosuperior iliac crests. On entry to the abdominal cavity, a wound retractor is placed to optimize access to the lower uterine segment so that a low transverse uterine incision can be made. We have used this incision technique on 17 obese patients with no incidence of intraoperative complications, no requirement to perform a vertical hysterotomy incision, and no postoperative wound infections. Our technique is a feasible alternative incision for obese parturients that avoids making a subpannicular Pfannenstiel incision and still allows for good access to the lower uterine segment.


Randomized clinical trial of rifampin-trimethoprim and sulfamethoxazole-trimethoprim in the treatment of localized urinary tract infections.
To investigate whether 10 days of rifampin-trimethoprim (RIF-TMP) or 6 weeks of sulfamethoxazole-trimethoprim (SMX-TMP) would decrease the relapse rate in patients with acute uncomplicated upper urinary tract infections in comparison with 10 days of SMX-TMP, we randomized 189 patients to receive RIF-TMP or SMX-TMP in a ratio of 1:2. After the site of infection was established by the antibody-coated bacterium (ACB) test, patients with upper-tract infections who received SMX-TMP were again randomized and received either a total of 6 weeks or 10 days of therapy. All patients who received RIF-TMP were treated for 10 days. Clinical and microbiological evaluations were repeated at 2 and 6 weeks posttreatment. Eighty-five patients (54 ACB positive) received 10 days of RIF-TMP, 71 patients (45 ACB positive) received 10 days of SMX-TMP, and 18 patients (18 ACB positive) received 6 weeks of SMX-TMP. The overall recurrence rates in patients who received 10 days of therapy were 32% for RIF-TMP and 23% for SMX-TMP (P = 0.13). There were 12 (14%) relapses in the RIF-TMP group compared with 2 (3%) relapses in the SMX-TMP group (P = 0.01). In patients with upper-tract infections, the relapse rates were not statistically significantly different (P = 0.13). There were two (11%) recurrences (one relapse and one reinfection) in the 6-week treatment group. This 6% relapse rate was not different from the 4% relapse rate observed in patients with upper-tract infections who received 10 days of SMX-TMP. The number of patients who discontinued treatment because of an adverse effect in the 6-week SMX-TMP treatment group was significantly greater than those in the 10-day SMX-TMP treatment group (P=0.003) and the RIF-TMP treatment group (P=0.05). Ten days of SMX-TMP was as effective as 6 weeks of SMP-TMP or 10 days of RIF-TMP in the treatment of uncomplicated upper urinary tract infections and caused the fewest untoward effects.


Neuroproteomics and systems biology-based discovery of protein biomarkers for traumatic brain injury and clinical validation.
The rapidly growing field of neuroproteomics has expanded to track global proteomic changes underlying various neurological conditions such as traumatic brain injury (TBI), stroke, and Alzheimer's disease. TBI remains a major health problem with approximately 2 million incidents occurring annually in the United States, yet no affective treatment is available despite several clinical trials. The absence of brain injury diagnostic biomarkers was identified as a significant road-block to therapeutic development for brain injury. Recently, the field of neuroproteomics has undertaken major advances in the area of neurotrauma research, where several candidate markers have been identified and are being evaluated for their efficacy as biological biomarkers in the field of TBI. One scope of this review is to evaluate the current status of TBI biomarker discovery using neuroproteomics techniques, and at what stage we are at in their clinical validation. In addition, we will discuss the need for strengthening the role of systems biology and its application to the field of neuroproteomics due to its integral role in establishing a comprehensive understanding of specific brain disorder and brain function in general. Finally, to achieve true clinical input of these neuroproteomic findings, these putative biomarkers should be validated using preclinical and clinical samples and linked to clinical diagnostic assays including ELISA or other high-throughput assays.


High-efficiency BODIPY-based organic photovoltaics.
A benzannulated boron dipyrromethene (BODIPY, bDIP) molecule exhibiting strong absorption at 640 nm was synthesized. The organic dye was used in an organic solar cell as the electron donor with C60 as the acceptor. The BODIPY dye demonstrated the best performance in lamellar architecture (indium tin oxide (ITO)/bDIP/C60/bathocuproine/Al), giving power conversion efficiency up to 4.5% with short-circuit current (JSC) of 8.7 mA/cm(2) and an open-circuit voltage (VOC) of 0.81 V. Neutron reflectivity experiments were performed on the bilayer film to investigate the thickness dependence of JSC. A 13 nm mixed layer was found to be present at the donor/acceptor interface in the bilayer device, formed when the C60 was deposited onto a room temperature bDIP film. Planar-mixed heterojunction devices were fabricated to understand the extent of spontaneous mixing between the donor and acceptor materials. The native mixed region in the bilayer device was shown to most resemble 1:3 bDIP:C60 layer in the structure: (ITO/bDIP/bDIP:C60 blend/C60/bathocuproine/Al).


A quartz crystal microbalance study of polycation-supported single and double stranded DNA surfaces.
A quartz crystal microbalance with dissipation monitoring (QCM-D) was used to investigate the properties and formation of a genomic mammalian DNA surface on a polycationic poly(ethylenimine) (PEI) film. We show that both single- and double-stranded DNA films can be deposited on the PEI surface by modulating the DNA adsorption time. The two distinct DNA surfaces can be confirmed by their interactions with urea, a common DNA denaturant, and ethidium bromide, a common DNA intercalator, both of which lead to characteristic changes in the QCM-D frequency and dissipation. The hybridization process between surface-bound single-stranded DNA to complementary strands in solution can be resolved in real-time. Moreover, we have also investigated the effects of incorporating NaCl in the various PEI-DNA assemblies and have shown that higher ionic strengths lead to greater DNA adsorption to the PEI surface. An increase in the QCM-D resonant frequency and a decrease in dissipation occur when these assemblies are rinsed with salt-free water. We interpret these changes as a loss of counterions from the film and an increase in intrinsic ion-pair complexation, leading to a more rigid PEI-DNA assembly. Varying the salt content in the DNA film can be used to control the film thickness and morphology.


Association between serum and oral matrix metalloproteinase-8 levels and periodontal health status.
Matrix metalloproteinase-8 (MMP-8) is involved in a wide range of pathologies including periodontitis and cardiovascular diseases (CVD). The association between periodontitis and CVD has been repeatedly recognized. The aim of the study was to analyze to what extent circulating active MMP-8 (aMMP-8) is associated with periodontal disease status and oral fluid aMMP-8 levels in otherwise healthy subjects. In a cross-sectional study, aMMP-8 was measured in serum of 59 volunteers, comprising 19 periodontally healthy subjects, 20 patients with gingivitis as well as 20 with periodontitis. All study subjects were characterized regarding aMMP-8 concentrations in different oral fluids as well as clinically and microbiologically with respect to periodontal disease. aMMP-8 levels in gingival crevicular fluid were measured using the enzyme-linked immunosorbent assay. Saliva enzyme levels as well as circulating aMMP-8 were determined by a time-resolved immunofluorometric assay. Both methods utilized the same monoclonal antibodies. Correlation and regression analyses were performed to study the potential association between serum aMMP-8 and oral parameters. Oral aMMP-8 levels were significantly higher in patients with periodontitis compared to periodontally healthy or gingivitis subjects. Highest serum aMMP-8 concentration was also found in the periodontitis group. The serum levels correlated significantly with oral aMMP-8 as well as with clinical parameters in a dose-dependent manner. These results were confirmed in a multivariate regression analysis. After adjusting for potential confounders, saliva aMMP-8 concentrations as well as periodontitis severity were significant predictors of serum aMMP-8. The associations between circulating aMMP-8 and oral aMMP-8 as well as periodontal findings in a dose-dependent manner may contribute to linking periodontal disease with increased CVDsusceptibility.


Influenza A Virus Utilizes Low-Affinity, High-Avidity Interactions with the Nuclear Import Machinery To Ensure Infection and Immune Evasion.
The incoming influenza A virus (IAV) genome must pass through two distinct barriers in order to establish infection in the cell: the plasma membrane and the nuclear membrane. A precise understanding of the challenges imposed by the nuclear barrier remains outstanding. Passage across is mediated by host karyopherins (KPNAs), which bind to the viral nucleoprotein (NP) via its N-terminal nuclear localization sequence (NLS). The binding affinity between the two molecules is low, but NP is present in a high copy number, which suggests that binding avidity plays a compensatory role during import. Using nanobody-based technology, we demonstrate that a high binding avidity is required for infection, though the absolute value differs between cell types and correlates with their relative susceptibility to infection. In addition, we demonstrate that increasing the affinity level caused a decrease in avidity requirements for some cell types but blocked infection in others. Finally, we show that genomes that become frustrated by low avidity and remain cytoplasmic trigger the type I interferon response. Based on these results, we conclude that IAV balances affinity and avidity considerations in order to overcome the nuclear barrier across a broad range of cell types. Furthermore, these results provide evidence to support the long-standing hypothesis that IAV's strategy of import and replication in the nucleus facilitates immune evasion.IMPORTANCE We used intracellular nanobodies to block influenza virus infection at the step prior to nuclear import of its ribonucleoproteins. By doing so, we were able to answer an important but outstanding question that could not be addressed with conventional tools: how many of the ∼500 available NLS motifs are needed to establish infection? Furthermore, by controlling the subcellular localization of the incoming viral ribonucleoproteins and measuring the cell's antiviral response, we were able to provide direct evidence for the long-standing hypothesis that influenza virus exploits nuclear localization to delay activation of the innate immune response.


Synthesis of aryl, glycosyl, and azido septanosides through ring expansion of 1,2-cyclopropanated sugars.
A ring-expansion methodology for the preparation of aryl septanosides, arabinofuranosyl and glucopyranosyl septanoside disaccharides, and azido septanosides is reported. A cyclopropanated adduct of the oxyglycal upon reaction with phenols, sugars, and azide led to the formation of ring-expanded septanoside derivatives. The ring expansion was found to be stereoselective with sugars, whereas phenols and the azide afforded an anomeric mixture of the ring expanded product. It was observed further that the conversion of the intermediate diketones to the diols, using NaBH(4), occurred with high diastereoselectivities for the alpha-anomers of the septanosides. This report consolidates further the generality of the oxyglycal ring-expansion method to prepare septanosides, possessing different substituents at their reducing ends.


Toll-like receptor-2 exacerbates murine acute viral hepatitis.
Viral replication in the liver is generally detected by cellular endosomal Toll-like receptors (TLRs) and cytosolic helicase sensors that trigger antiviral inflammatory responses. Recent evidence suggests that surface TLR2 may also contribute to viral detection through recognition of viral coat proteins but its role in the outcome of acute viral infection remains elusive. In this study, we examined in vivo the role of TLR2 in acute infections induced by the highly hepatotrophic mouse hepatitis virus (MHV) type 3 and weakly hepatotrophic MHV-A59 serotype. To address this, C57BL/6 (wild-type; WT) and TLR2 knockout (KO) groups of mice were intraperitoneally infected with MHV3 or MHV-A59. MHV3 infection provoked a fulminant hepatitis in WT mice, characterized by early mortality and high alanine and aspartate transaminase levels, histopathological lesions and viral replication whereas infection of TLR2 KO mice was markedly less severe. MHV-A59 provoked a comparable mild and subclinical hepatitis in WT and TLR2 KO mice. MHV3-induced fulminant hepatitis in WT mice correlated with higher hepatic expression of interferon-β, interleukin-6, tumour necrosis factor-α, CXCL1, CCL2, CXCL10 and alarmin (interleukin-33) than in MHV-A59-infected WT mice and in MHV3-infected TLR2 KO mice. Intrahepatic recruited neutrophils, natural killer cells, natural killer T cells or macrophages rapidly decreased in MHV3-infected WT mice whereas they were sustained in MHV-A59-infected WT mice and MHV3-infected TLR2 KO. MHV3 in vitro infection of macrophagic cells induced rapid and higher viral replication and/or interleukin-6 induction in comparison to MHV-A59, and depended on viral activation of TLR2 and p38 mitogen-activated protein kinase. Taken together, these results support a new aggravating inflammatory role for TLR2 in MHV3-induced acute fulminant hepatitis.


Formulation and Evaluation of a Self-microemulsifying Drug Delivery System Containing Bortezomib.
The purposes of the present study were to develop a self-microemulsifying drug delivery system (SMEDDS) containing bortezomib, a proteasome inhibitor. The solubility of the drug was evaluated in 15 pharmaceutical excipients. Combinations of oils, surfactants and cosurfactants were screened by drawing pseudo-ternary phase diagrams. The system exhibiting the largest region of microemulsion was considered optimal. Bortezomib SMEDDS spontaneously formed a microemulsion when diluted with an aqueous medium with a median droplet size of approximately 20-30 nm. In vitro release studies showed that the SMEDDS had higher initial release rates for the drug when compared with the raw drug material alone. Measurement of the viscosity, size, and ion conductivity indicated that a phase inversion from water in an oil system to oil in a water system occurred when the weight ratio of the water exceeded 30% of the entire microemulsion system. In a pharmacokinetics study using rats, the bortezomib microemulsion failed to improve the bioavailability of the drug. The reason was assumed to be degradation of the drug in the microemulsion in the gastrointestinal tract. However, bortezomib in Labrasol(®) solution (an aqueous solution containing 0.025% Labrasol(®)) showed significantly increased area under the curve from 0-24 h (AUC0-24 h) and maximum plasma concentration (Cmax) values compared to the drug suspension. The findings of this study imply that oral delivery of a bortezomib and colloidal system containing Labrasol(®) could be an effective strategy for the delivery of bortezomib.


Online Conductimetric Flow-Through Analyzer Based on Membrane Diffusion for Ammonia Control in Wastewater Treatment Process.
Ammonia is a necessary monitoring parameter that should be controlled within an optimum range in the whole process of wastewater treatment and recycling, but few reliable real-time monitoring technologies are available currently under such harsh water conditions. This study proposes a continuous conductometric flow-through analyzer for ammonia monitoring (CFAA) in the wastewater treatment process. It is developed based on the gas diffusion mechanisms, and the proposed analytical principle has been validated in which the real-time conductivity increment rate is linearly proportional to the real-time ammonia concentration in the sample. This method could be generally applicable in monitoring a wide ammonia concentration range (10.2 μg L-1 to 500 mg L-1), and it is capable of achieving long-term ammonia monitoring by periodic renewal of the receiving solution. The potential impact factors and corresponding calibration principles are also developed to avoid tedious ongoing calibration. The field application results demonstrate that CFAA can effectively and directly achieve real-time and average ammoniacal nitrogen monitoring at different treatment stages regardless of the complexity of wastewater, not requiring any sample pretreatment. Compared with other ammonia online monitoring technologies, the proposed CFAA shows remarkable advantages in high reliability, durability, and accuracy, especially under severe monitoring condition. It can be a useful monitoring tool for continuous ammonia control in the wastewater treatment process.


Pravastatin Inhibits the Rho/CCN2/extracellular matrix cascade in human fibrosis explants and improves radiation-induced intestinal fibrosis in rats.
Intestinal complications after radiotherapy are caused by transmural fibrosis and impair the quality of life of cancer survivors. Radiation fibrosis was considered permanent and irreversible, but recently, its dynamic nature was shown, providing new opportunities for the development of antifibrotic therapies. Among these new targets, we identified the Rho/ROCK pathway and thought to investigate whether pravastatin treatment inhibits Rho pathway activation and elicits an antifibrotic action. Rho and ROCK activities were monitored in human explants presenting radiation fibrosis remodeling after incubation with pravastatin. Subsequent modulation of CCN2, type I collagen, and fibronectin expression were assessed ex vivo and in intestinal smooth muscle cells derived from radiation enteropathy. Then, the therapeutic relevance of the antifibrotic action of pravastatin was explored in vivo in a rat model of chronic radiation fibrosis (19 Gy X-rays) treated with 30 mg/kg/d pravastatin in the drinking water. The results obtained with human explants show that pravastatin specifically inhibits Rho activity in submucosal mesenchymal cells. Pravastatin also elicits ROCK inhibition, and subsequent CCN2 production in human explants and smooth muscle cells isolated from radiation enteropathy. Inhibition of type I collagen and fibronectin does occur, showing that pravastatin modulates the secretory phenotype of mesenchymal cells. Lastly, curative pravastatin administration improves radiation enteropathy in rats. This structural improvement is associated with decreased deposition of CCN2 and subsequent decreased extracellular matrix deposition. Targeting established fibrosis with pravastatin is an efficient and safe antifibrotic strategy in radiation-induced enteropathy, and is easily transferable into the clinic.


A novel necroptosis-related lncRNA signature predicts the prognosis and immune microenvironment of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is one of the malignant tumors with high mortality and a worse prognosis globally. Necroptosis is a programmed death mediated by receptor-interacting Protein 1 (RIP1), receptor-interacting Protein 1 (RIP3), and Mixed Lineage Kinase Domain-Like (MLKL). Our study aimed to create a new Necroptosis-related lncRNAs (NRlncRNAs) risk model that can predict survival and tumor immunity in HCC patients. The RNA expression and clinical data originated from the TCGA database. Pearson correlation analysis was applied to identify the NRlncRNAs. The LASSO-Cox regression analysis was employed to build the risk model. Next, the ROC curve and the area under the Kaplan-Meier curve were utilized to evaluate the accuracy of the risk model. In addition, based on the two groups of risk model, we performed the following analysis: clinical correlation, differential expression, PCA, TMB, GSEA analysis, immune cells infiltration, and clinical drug prediction analysis. Plus, qRT-PCR was applied to test the expression of genes in the risk model. Finally, a prognosis model covering six necroptosis-related lncRNAs was constructed to predict the survival of HCC patients. The ROC curve results showed that the risk model possesses better accuracy. The 1, 3, and 5-years AUC values were 0.746, 0.712, and 0.670, respectively. Of course, we also observed that significant differences exist in the following analysis, such as functional signaling pathways, immunological state, mutation profiles, and medication sensitivity between high-risk and low-risk groups of HCC patients. The result of qRT-PCR confirmed that three NRlncRNAs were more highly expressed in HCC cell lines than in the normal cell line. In conclusion, based on the bioinformatics analysis, we constructed an NRlncRNAs associated risk model, which predicts the prognosis of HCC patients. Although our study has some limitations, it may greatly contribute to the treatment of HCC and medical progression.


Comparison of ring 1 parameters in 37-segment multifocal electroretinography between onset and offset conditions of ring 2 to 4 in normal subjects.
To investigate whether the response of a central hexagonal element corresponding to the macular area in conventional multifocal electroretinography (mfERG) tests was the same as that of experimental mfERG using single central hexagonal element stimulation. Prospective, observational study. Thirty healthy subjects were included in this study. mfERG recordings were performed according to two protocols: stimulus with 37 hexagonal elements (protocol 1), and stimulus with a single central element created by deactivating the other 36 hexagonal elements (protocol 2). We compared differences between ring 1 parameters in each protocol. In protocol 1, the first positive component (P1) implicit time and P1 amplitude were 37.8±1.8ms and 6.3±2.7 µV. After single element stimulation (protocol 2), double positive waves appeared. The implicit time and amplitude of P1 were 40.7±2.4ms (P<0.001) and 9.1±3.3 µV (P=0.001), respectively. The implicit time and amplitude of the second positive component (P2) were 68.0±4.5ms (P<0.001, compared with P1 in protocol 1) and 12.3±4.7 µV (P<0.001, compared with P1 in protocol 1), respectively. The amplitude of P2 in protocol 2 was about two times higher than that of P1 in protocol 1. mfERG responses of a central hexagonal element in a single element stimulation protocol are different from those of multiple element stimulation. The positive wave is more enhanced compared to that of the conventional protocol and it elongated into two wavelets.


The Hispanic Clinic for Pediatric Surgery: A model to improve parent-provider communication for Hispanic pediatric surgery patients.
26 million Americans have limited English proficiency (LEP). It is well established that language barriers adversely affect health and health care. Despite growing awareness of language barriers, there is essentially a void in the medical literature regarding the influence of language disparity on pediatric surgery patients. This study was designed to assess the impact of patient-provider language concordance on question-asking behavior and patient satisfaction for pediatric surgery patients. Participants included families of patients in a General Pediatric Surgery Clinic categorized into 3 groups by patient-provider language concordance: concordant English-speaking, LEP concordant Spanish-speaking, and LEP discordant Spanish-speaking using an interpreter. Clinical visits were audio recorded and the number of patient-initiated questions and the length of clinical encounter were measured. Families were administered a surgery-specific, 5-point Likert scale questionnaire modeled after validated surveys concerning communication, trust, perceived discrimination and patient-provider language concordance. Regression models were performed to analyze associations between language concordance and patient's question-asking behavior and between language concordance and survey results. A total of 156 participants were enrolled including 57 concordant-English, 52 LEP concordant-Spanish and 47 LEP-discordant-Spanish. There was significant variation in the mean number of patient-initiated questions among the groups (p=0.002). Both the English and Spanish concordant groups asked a similar number of questions (p=0.9), and they both asked more questions compared to the Spanish-discordant participants (p=0.002 and p=0.001). Language discordance was associated with fewer questions asked after adjustment for socioeconomic status. Language concordant participants rated higher scores of communication. Both Spanish-concordant and Spanish-discordant patients reported significantly increased preference for, and value of language concordant care. Language discordant participants reported that they desired to ask more questions but were limited by a language barrier (p=0.001). In a pediatric surgery clinic, language concordant care increases the number of patient-asked questions during a clinical visit and improves communication suggesting that discordant care is a potential source of disparities in access to information. Future efforts should focus on expanding access to language concordant providers in other surgery subspecialties as a step towards limiting disparities in surgical care for all patients.


Euthanasia and the quality of life debate.
Orthodox Christian ethics is grounded in the sacredness of life principle. Yet, it can accept a quality of life approach where "quality" refers not to capacities or states, but to the relationship between the patient's condition and the quest for transcendent life goals (Walter and Shannon, 1990). The true quality of human life derives from the vocation to stewardship, which enjoins an attitude of humble acceptance toward beneficial or "redemptive" suffering. The proper response to suffering in terminal cases is not active euthanasia or physician-assisted suicide but appropriate pain management and personal care. In cases of PVS or deep coma, only the determination of higher brain death can warrant the withholding or withdrawing of food and hydration. Yet, artificial maintenance of biological existence is also immoral. Death is to be accepted and embraced as a transition to eternal life.


Ventilatory responses to the metabolic acidosis of treadmill and cycle ergometry.
Ventilation and acid-base responses were studied at comparable levels of O2 uptake during cycle ergometer and treadmill exercise, to determine the extent to which the type of exercise affects these responses. Twenty male subjects performed 50-, 100-, and 150-W cycle ergometer exercise and three work rates of similar O2 uptake on a treadmill. At comparable oxygen uptakes, arterial lactate and VE were higher and arterial pH and bicarbonate were lower for cycle ergometer than treadmill exercise. These differences could be accounted for by the greater degree of metabolic acidosis during cycle ergometer work. The increment in VE over that predicted (from an extrapolation of the linear relationship of the VE-VO2 relationship for low work rates) was linearly related to the decrease in arterial bicarbonate; VE was increased by approximately 4 1/min for each meq/1 of bicarbonate decrease for both treadmill and cycle ergometry.


Use of AlloDerm implant to improve cosmesis after parotidectomy.
We evaluated the effectiveness and practicality of using AlloDerm, an acellular human dermal matrix graft, as an interpositional barrier in an attempt to improve the appearance of the surgical defect created by parotidectomy. We performed AlloDerm reconstruction in a series of 10 patients, and we found that normal contour was satisfactorily restored in all 10. We conclude that the use of an AlloDerm implant is a low-risk, practical option for repairing the surgical defect in postparotidectomy patients.


A review of spatial capture-recapture: Ecological insights, limitations, and prospects.
First described by Efford (2004), spatial capture-recapture (SCR) has become a popular tool in ecology. Like traditional capture-recapture, SCR methods account for imperfect detection when estimating ecological parameters. In addition, SCR methods use the information inherent in the spatial configuration of individual detections, thereby allowing spatially explicit estimation of population parameters, such as abundance, survival, and recruitment. Paired with advances in noninvasive survey methods, SCR has been applied to a wide range of species across different habitats, allowing for population- and landscape-level inferences with direct consequences for conservation and management. I conduct a literature review of SCR studies published since the first description of the method and provide an overview of their scope in terms of the ecological questions answered with this tool, taxonomic groups targeted, geography, spatio-temporal extent of analyses, and data collection methods. In addition, I review approaches for analytical implementation and provide an overview of parameters targeted by SCR studies and conclude with current limitations and future directions in SCR methods.


Nucleophosmin mutations in acute myeloid leukemia: a tale of protein unfolding and mislocalization.
Nucleophosmin (NPM1) is an abundant, ubiquitously expressed protein mainly localized at nucleoli but continuously shuttling between nucleus and cytoplasm. NPM1 plays a role in several cellular functions, including ribosome biogenesis and export, centrosome duplication, chromatin remodeling, DNA repair, and response to stress stimuli. Much of the interest in this protein arises from its relevance in human malignancies. NPM1 is frequently overexpressed in solid tumors and is the target of several chromosomal translocations in hematologic neoplasms. Notably, NPM1 has been characterized as the most frequently mutated gene in acute myeloid leukemia (AML). Mutations alter the C-terminal DNA-binding domain of the protein and result in its aberrant nuclear export and stable cytosolic localization. In this review, we focus on the leukemia-associated NPM1 C-terminal domain and describe its structure, function, and the effect exerted by leukemic mutations. Finally, we discuss the possibility to target NPM1 for the treatment of cancer and, in particular, of AML patients with mutated NPM1 gene.


A reverse hemolytic plaque assay for microscopic visualization of growth hormone release from individual cells: evidence for somatotrope heterogeneity.
Growth hormone (GH) secreting cells direct complement-mediated plaque formation (clear zones of hemolysis surrounding the somatotropes) in mixed pituitary cell cultures incubated as a monolayer with protein-A coupled ovine erythrocytes (oRBC) in the presence of GH antiserum. Plaques were maximal in number after 4 h; growth hormone-releasing hormone (GHRH) and somatostatin increased and decreased, respectively, the rate of formation of plaques and their final sizes. Approximately 30% of all pituitary cells formed GH plaques and a similar fraction stained for GH using peroxidase-antiperoxidase immunocytochemistry (ICC). The plaque areas of individual somatotropes (reflecting the amount of GH released) covered a 20-fold range from the smallest to the largest in the 3 treatment groups. Somatostatin-treated and untreated cells formed frequency distributions of plaque sizes that were unimodal. In contrast, GHRH produced a bimodal frequency distribution suggestive of a sub-population of somatotropes preferentially responsive to this secretagogue. This new assay coupled with other morphological and biochemical techniques that can be applied to single cells will permit further analysis of these sub-populations of somatotropes.


Nationwide evaluation of malaria infections, morbidity, mortality, and coverage of malaria control interventions in Madagascar.
In the last decade, an important scale-up was observed in malaria control interventions. Madagascar entered the process for pre-elimination in 2007. Policy making needs operational indicators, but also indicators about effectiveness and impact of malaria control interventions (MCI). This study is aimed at providing data about malaria infection, morbidity, and mortality, and MCI in Madagascar. Two nationwide surveys were simultaneously conducted in 2012-2013 in Madagascar: a study about non-complicated clinical malaria cases in 31 sentinel health facilities, and a cross-sectional survey (CSS) in 62 sites. The CSS encompassed interviews, collection of biological samples and verbal autopsies (VA). Data from CSS were weighted for age, sex, malaria transmission pattern, and population density. VA data were processed with InterVA-4 software. CSS included 15,746 individuals of all ages. Parasite rate (PR) as measured by rapid diagnostic tests was 3.1%, and was significantly higher in five to 19 year olds, in males, poorer socio-economic status (SES) quintiles and rural areas. Long-lasting insecticidal nets (LLIN) use was 41.7% and was significantly lower in five to 19 year olds, males and wealthier SES quintiles. Proportion of persons covered by indoor residual spraying (IRS) was 66.8% in targeted zones. Proportion of persons using other insecticides than IRS was 22.8%. Coverage of intermittent preventive treatment during pregnancy was 21.5%. Exposure to information, education and communication messages about malaria was significantly higher in wealthier SES for all media but information meetings. The proportion of fever case managements considered as appropriate with regard to malaria was 15.8%. Malaria was attributed as the cause of death in 14.0% of 86 VA, and 50% of these deaths involved persons above the age of five years. The clinical case study included 818 cases of which people above the age of five accounted for 79.7%. In targeted zones, coverage of LLIN and IRS were lower in clinical cases than in general population. This study provides valuable data for the evaluation of effectiveness and factors affecting MCI. MCI and evaluation surveys should consider the whole population and not only focus on under-fives and pregnant women in pre-elimination or elimination strategies.


Role of toll-like receptors and nod-like receptors in acute lung infection.
The respiratory system exposed to microorganisms continuously, and the pathogenicity of these microbes not only contingent on their virulence factors, but also the host's immunity. A multifaceted innate immune mechanism exists in the respiratory tract to cope with microbial infections and to decrease tissue damage. The key cell types of the innate immune response are macrophages, neutrophils, dendritic cells, epithelial cells, and endothelial cells. Both the myeloid and structural cells of the respiratory system sense invading microorganisms through binding or activation of pathogen-associated molecular patterns (PAMPs) to pattern recognition receptors (PRRs), including Toll-like receptors (TLRs) and NOD-like receptors (NLRs). The recognition of microbes and subsequent activation of PRRs triggers a signaling cascade that leads to the activation of transcription factors, induction of cytokines/5chemokines, upregulation of cell adhesion molecules, recruitment of immune cells, and subsequent microbe clearance. Since numerous microbes resist antimicrobial agents and escape innate immune defenses, in the future, a comprehensive strategy consisting of newer vaccines and novel antimicrobials will be required to control microbial infections. This review summarizes key findings in the area of innate immune defense in response to acute microbial infections in the lung. Understanding the innate immune mechanisms is critical to design host-targeted immunotherapies to mitigate excessive inflammation while controlling microbial burden in tissues following lung infection.


Chemical accidents in freshwater: Development of forecasting system for drinking water resources.
Chemical accidents have threatened drinking water safety and aquatic systems when hazardous chemicals flow into inland waterbodies through pipelines in industrial complexes. In this study, a forecasting system was developed for the prevention of drinking water resource pollution by considering chemical transport/fate through both pipelines and river channels. To this end, we coupled a pipe network model (Storm Water Management Model) with a calibrated hydrodynamic model (Environmental Fluid Dynamics Code). In addition, we investigated whether chemical transport through pipelines would make a difference in chemical concentration predictions. For both pipelines and river channels, the results showed lower peak concentrations than those without pipelines, whereas the time of peak concentration did not change significantly. When chemicals were transported with both pipelines and river channels, the peak concentrations were 25.81% and 41.91% lower than those of chemicals carried directly into the Han and Geum Rivers without the pipeline transport. Further, our system is automated from scenario generation to analysis and usage is straightforward, with a simple input of accident information. The results of this study can be utilized to establish a safe water supply system and preliminary countermeasures against accidental water pollution in the future.


Short communication: Identification of Corynebacterium bovis by MALDI-mass spectrometry.
Corynebacterium bovis is a mastitis-causing microorganism responsible for economic losses related to decrease in milk production. The aim of the study was identify Corynebacterium spp. strains recovered from milk samples of subclinical mastitis by using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Samples were collected during a 10-mo mastitis-monitoring program in a high-production dairy farm. In this study, 80 strains were analyzed; from these 54 (67.5%) were identified at species level as Corynebacterium bovis, 24 (31.2%) isolates were identified at the genus level as Corynebacterium spp., and only 1 (1.35%) isolated had unreliable identification. Results demonstrated that MALDI-MS could be an important technique for the identification of Corynebacterium spp. in milk.


Maternal and perinatal outcome among the booked and unbooked pregnancies from catchments area of BP Koirala Institute of Health Sciences, Nepal.
To examine the differences in maternal and perinatal outcome among booked and unbooked pregnant women and their perception for underutilization of services targeted for them. Two thousand two hundred and twenty eight pregnant women, booked and unbooked, attending emergency obstetrics care during Jan.-Dec. 2005, were included in the study; Investigators followed cases till discharge. Tendency for mothers to be booked was found to be significantly associated with age. Booked and unbooked mothers differed by distance to the hospital and parity. There was difference between the periods of gestation at which they presented to the hospital among both the groups (p<0.0001). Nearly eighty percent (81.4%) of unbooked mothers did not know the period of gestation. Most of these women conceived during lactational or depo provera-induced amenorrhea. The maternal mortality rate for unbooked mothers was 16 out of 1056 and none among the booked. Perinatal mortality is 3 times less in booked mothers. Higher perinatal mortalities are associated with unbooked mothers and had a higher proportion of maternal intensive care unit admissions and perinatal deaths compared with the booked group. Focus group discussion revealed that the reasons for underutilizations of services are; Distance, cost, disempowerment of women and attitudes of hospital personnel. The opportunities to enhance utilizations are based on distance, cost and client friendly environment of hospital personnel. It has been observed, building teaching hospitals alone, will not have much impact to maternal and perinatal outcome, unless peripheral health institutions are equipped with facilities and service provisions.


Direct α-Imination of N-Acyl Pyrazoles with Nitrosoarenes.
Unprecedented α-imino N-acyl pyrazoles were efficiently and selectively prepared through the 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD)-catalyzed reaction of nitrosoarenes with N-acyl pyrazoles via an N-nitroso aldol reaction/dehydration sequence. The α-imino acyl pyrazoles were demonstrated to be new versatile intermediates for practical one-pot syntheses of α-imino amides, dipeptide precursors, esters, and β-amino alcohols. The synthetic method competes with known protocols in terms of ready availability of the reagents and catalyst, mild and catalytic reaction conditions, gram-scale applicability, and scope of the α-imino acid derivatives achievable.


The Relation between Vaporization Enthalpies and Viscosities: Eyring's Theory Applied to Selected Ionic Liquids.
Key properties for the use of ionic liquids as electrolytes in batteries are low viscosities, low vapor pressure and high vaporization enthalpies. Whereas the measurement of transport properties is well established, the determination of vaporization enthalpies of these extremely low volatile compounds is still a challenge. At a first glance both properties seem to describe different thermophysical phenomena. However, eighty years ago Eyring suggested a theory which related viscosities and vaporization enthalpies to each other. The model is based on Eyring's theory of absolute reaction rates. Recent attempts to apply Eyring's theory to ionic liquids failed. The motivation of our study is to show that Eyring's theory works, if the assumptions specific for ionic liquids are fulfilled. For that purpose we measured the viscosities of three well selected protic ionic liquids (PILs) at different temperatures. The temperature dependences of viscosities were approximated by the Vogel-Fulcher-Tamann (VFT) relation and extrapolated to the high-temperature regime up to 600 K. Then the VFT-data could be fitted to the Eyring-model. The values of vaporization enthalpies for the three selected PILs predicted by the Eyring model have been very close to the experimental values measured by well-established techniques. We conclude that the Eyring theory can be successfully applied to the chosen set of PILs, if the assumption that ionic pairs of the viscous flow in the liquid and the ionic pairs in the gas phase are similar is fulfilled. It was also noticed that proper transfer of energies can be only derived if the viscosities and the vaporization energies are known for temperatures close to the liquid-gas transition temperature. The idea to correlate easy measurable viscosities of ionic liquids with their vaporization enthalpies opens a new way for a reliable assessment of these thermodynamic properties for a broad range of ionic liquids.


Left Ventricular Diastolic Dysfunction as Predictor of Unfavorable Prognosis After ESUS.
Identification of echocardiographic, hemodynamic and biochemical predictors of unfavorable prognosis after embolic strokes of undetermined etiology (ESUS) in patients at age <65. Out of 520 ischemic stroke patients we selected 64 diagnosed with ESUS and additional 36 without stroke but with similar risk profile. All patients underwent echocardiography, non-invasive assessment of hemodynamic parameters using SphygmoCor tonometer and measurements of selected biomarkers. Follow-up time was 12 months. Nine percent of patients died, and recurrent ischemic stroke occurred in 9% of patients only in the ESUS group. Atrial fibrillation (AF) occurred in 10% of patients and the ESUS group had a significantly poorer outcome of AF in the first 2 months after hospitalization. The outcome of re-hospitalization was 28% in the ESUS group and 17% in the control group. In the multivariate analysis mean early diastolic (E') mitral annular velocity (OR 0.75, 95% CI: 0.6-0.94; p=0.01) was significantly associated with cardiovascular hospitalizations. The only independent predictor of recurrent stroke was the ratio of peak velocity of early diastolic transmitral flow to peak velocity of early diastolic mitral annular motion (E/E') (OR 0.75, 95% CI: 0.6-0.94; p=0.01). E/E' was independently associated with composite endpoint (death, hospitalization and recurrent stroke) (OR 1.90, 95% CI 1.1-3.2, p=0.01). The indices of diastolic dysfunction are significantly associated with unfavorable prognosis after ESUS. There is a robust role for outpatient cardiac monitoring especially during the first 2 months after ESUS to detect potential AF.


Focused assessment with sonography in trauma (FAST): should its role be reconsidered?
Focused assessment with sonography in trauma (FAST) is a limited ultrasound scan performed in the emergency department to assess patients admitted with blunt abdominal trauma (BAT). It is normally undertaken by emergency physicians in order to identify the presence of free fluid, which may represent haemoperitoneum. This potentially allows prompt referral to further imaging, such as a computed tomography (CT) scan, and/or surgery. FAST has been adopted worldwide, and most major trauma centres now have access to an ultrasound machine. Despite the popularity of FAST, there remains a lack of clarity and evidence around any actual contribution to patient survival. An ability to evaluate the true impact of FAST is complicated by factors such as the operator dependence of ultrasound, the improved access, speed and subsequent use of CT (which has reduced the value of FAST in some centres), a lack of standardisation around education and the most effective methods to ensure competence, and variations in the practice, quality and audit of FAST. Enthusiasm for FAST is undiminished among emergency physicians, and in general, point of care ultrasound is escalating in a range of areas, including the emergency department. Small portable and hand held ultrasound machines improve the accessibility of FAST and its benefits are frequently perceived to be an established and essential part of the emergency department service despite its limitations. Since the introduction of FAST, trauma patients are increasingly managed by non-surgical means if haemodynamically stable, and the presence of intraperitoneal fluid does not necessarily influence this decision. Recent developments in multi-detector CT, and in the use of contrast enhanced ultrasound, have broadened the management options for trauma patients, and can support a faster diagnostic pathway than was previously possible. FAST is a limited triage tool, whose actual value in the diagnostic and treatment pathway of patients with BAT remains questionable, and more data are needed to justify its use in the light of these recent developments.


Design of combination biotherapy studies: future goals and challenges.
The recent large-scale production of biomodulators, also known as biologic response modifiers, made possible through recombinant DNA technology, offers the potential for significant advances in the treatment of cancer. The antitumor activity of these agents, such as interferons, interleukins, and tumor necrosis factor, have generated enthusiasm for further investigation. In an effort to improve response rates, combinations of these agents both with and without conventional therapies are currently being examined. Clinical trials have been conducted with various therapeutic combinations, including a biomodulator plus chemotherapy, combinations of different biomodulators, a biomodulator with concomitant chemotherapy and radiation, and multiple combinations of chemotherapies and biomodulators. These approaches are promising and some limited successes have been reported; however, the goal of increased anticancer activity without greater toxicities or antagonism between various agents is not always achieved. Synergism among active agents is not necessarily assured and quite unexpected and unpredictable toxicities have been noted. The studies to date suggest that important new therapies will emerge, but many questions have to be answered before the specific roles of these new treatments are defined.


Probing Dative and Dihydrogen Bonding in Ammonia Borane with Electronic Structure Computations and Raman under Nitrogen Spectroscopy.
Although ammonia borane is isoelectronic with ethane and they have similar structures, BH3NH3 exhibits rather atypical bonding compared to that in CH3CH3. The central bond in ammonia borane is actually a coordinate covalent or dative bond rather than the conventional covalent C-C bond in ethane where each atom donates one electron. In addition, strong intermolecular dihydrogen bonds can form between two or more ammonia borane molecules compared to the relatively weak dispersion forces between ethane molecules. As a result, ammonia borane's physical properties are very sensitive to the environment. For example, gas-phase and solid-state ammonia borane have very different BN bond lengths and BN stretching frequencies, which led to much debate in the literature. It has been demonstrated that the use of cluster models based on experimental crystal structures led to better agreement between theory and experiment. Here, we employ a variety of cluster models to track how the interaction energies, bond lengths, and vibrational normal modes evolve with the size and structural characteristics of the clusters. The M06-2X/6-311++G(2df,2pd) level of theory was selected for this analysis on the basis of favorable comparison with CCSD(T)/aug-cc-pVTZ data for the ammonia borane monomer and dimer. Fourteen unique fully optimized molecular cluster geometries, (BH3NH3)n≤12, and nine crystal models, (BH3NH3)n≤19, were used to elucidate how the local environment impacts ammonia borane's physical properties. Computational results for the BN stretching frequencies are also compared directly to the Raman spectrum of solid ammonia borane at 77 K using Raman under liquid nitrogen spectroscopy (RUNS). A strong linear correlation was found to exist between the BN bond length and stretching frequency, from an isolated monomer to the most distorted BH3NH3 unit in a cluster or crystal structure model. Excellent agreement was seen between the frequencies computed for the largest crystal model and the RUNS experimental spectra (typically within a few wavenumbers).


Electron tunneling of photochemical reactions on metal surfaces: nonequilibrium Green's function-density functional theory approach to photon energy dependence of reaction probability.
We have developed a theoretical model of photoinduced reactions on metal surfaces initiated by the substrate/indirect excitation mechanism using the nonequilibrium Green's function approach. We focus on electron transfer, which consists of (1) electron-hole pair creation, (2) transport of created hot electrons, and (3) tunneling of hot electrons to form an anion resonance. We assume that steps (1), (2), and (3) are separable. By this assumption, the electron dynamics might be restated as a tunneling problem of an open system. Combining the Keldysh time-independent formalism with the simple transport theory introduced by Berglund and Spicer, we present a practical scheme for first-principle calculation of the reaction probability as a function of incident photon energy. The method is illustrated by application to the photoinduced desorption/dissociation of O2 on a Ag(110) surface by adopting density functional theory.


In vivo anti-metastatic effects of uPAR retargeted measles virus in syngeneic and xenograft models of mammary cancer.
The urokinase receptor (uPAR) plays a critical role in breast cancer (BC) progression and metastases and is a validated target for novel therapies. The current study investigates the effects of MV-uPA, an oncolytic measles virus fully retargeted against uPAR in syngeneic and xenograft BC metastases models. In vitro replication and cytotoxicity of MVs retargeted against human (MV-h-uPA) or mouse (MV-m-uPA) uPAR were assessed in human and murine cancer and non-cancer mammary epithelial cells. The in vivo effects of species-specific uPAR retargeted MVs were assessed in syngeneic and xenograft models of experimental metastases, established by intravenous administration of luciferase expressing 4T1 or MDA-MD-231 cells. Metastases progression was assessed by in vivo bioluminescence imaging. Tumor targeting was evaluated by qRT-PCR of MV-N, rescue of viable viral particles, and immunostaining of MV particles in lungs from tumor bearing mice. In vitro, MV-h-uPA and MV-m-uPA selectively infected, replicated, and induced cytotoxicity in cancer compared to non-cancer cells in a species-specific manner. In vivo, MV-m-uPA delayed 4T1 lung metastases progression and prolonged survival. These effects were associated with identification of viable viral particles, viral RNA, and detection of MV-N by immunostaining from lung tissues in treated mice. In the human MDA-MB-231 metastases model, intravenous administration of MV-h-uPA markedly inhibited metastases progression and significantly improved survival, compared to controls. No significant treatment-related toxicity was observed in treated mice. The above preclinical findings strongly suggest that uPAR retargeted measles virotherapy is a novel and feasible systemic therapy strategy against metastatic breast cancer.


[Monitoring ventilatory mechanics].
Monitoring the mechanical properties (both static and dynamic) of the respiratory tract is essential for the diagnosis and prognosis of the disease causing any condition and to take the pertinent therapeutic decisions. These measurements can maintain a lung function in optimum conditions during mechanical ventilation and choose the best time to disconnect the patient. Until recently, monitoring the respiration of patients undergoing mechanical ventilation required sophisticated and complicated apparatuses or difficult techniques. Instrumentalization of the respiratory as a monitor has evolved in such a way that many respirators can graphically reproduce in real time the changes that occur in some variables of the respiratory cycle. Using these monitoring techniques improves knowledge of lung behavior of each patient with ARDS, thus facilitating its management. It may also improve decision-making in each individual patient, improving their prognosis.


The genetic epidemiology of breast cancer genes.
Genetic susceptibility to breast cancer in women is conferred by a large number of genes, of which six have so far been identified. In the context of multiple-case families, BRCA1 and BRCA2 are the most important. Mutations in these genes confer high lifetime risks of breast cancer and ovarian cancer, and more moderate risks of prostate cancer and some other cancer types. Mutations in the CHEK2 and ATM genes, by contrast, cause much more modest (2-4 fold) risks of breast cancer. Genes so far identified explain approximately 20% of the familial aggregation of breast cancer. The remaining susceptibility genes have, so far, proved illusive, suggesting that they are numerous and confer moderate risks. A variety of techniques including genome-wide association studies, use of quantitative intermediate endpoints, and resequencing of genes may be required to identify them. The identification of such genes can provide a basis for targeted prevention of breast cancer.


Benefit of adding pioglitazone to successful statin therapy in nondiabetic patients with coronary artery disease.
Pioglitazone added to successful statin therapy may be beneficial for coronary artery disease (CAD) patients without diabetes mellitus. Fourteen individuals optimally treated with statin for >6 months were randomized to a pioglitazone or control group. Pioglitazone significantly improved insulin resistance, reduced high-sensitivity C-reactive protein, increased high-molecular-weight adiponectin and high-density lipoprotein cholesterol levels. Ultrasound echogenicity of carotid atheroma assessed by integrated backscatter was significantly increased by pioglitazone and correlated with adiponectin levels. Adding pioglitazone to successful statin therapy may be an effective therapeutic strategy for patients with CAD.


The occurrence and pathology of chlamydiosis in the male reproductive tract of non-human mammals: A review.
Organisms belonging to the Family Chlamydiaceae are responsible for a broad range of diseases in humans, livestock, companion animals and non-domestic species. Infection of the reproductive organs can cause a range of syndromes of which sub- and infertility are the most frequently observed clinical manifestations. While the gross and histological lesions associated with the isolation of Chlamydiaceae from the non-human female reproductive tract are well documented, little attention has been given to the pathological effects of this infection in the male genital system. As such, the occurrence and importance of Chlamydia-associated disease in male non-human mammalian species is less well documented. In order to improve our understanding of the significance of chlamydiosis in domestic, laboratory and wild animals, this review provides an up-to-date summary of Chlamydia-associated male reproductive pathology, whether that infection occurs naturally or experimentally. Although most lesions in males are described as incidental and of minor significance, results of recent studies suggest that infection with Chlamydiaceae can adversely impact male fertility and/or be instrumental in disease transmission. Although in humans, bulls and mice Chlamydia infection has been associated with morphological and functional abnormalities of the spermatozoa, this review will focus on the gross and histological findings linked to the colonisation of the genital system by this pathogen. Advances in our understanding of male reproductive chlamydiosis are necessary for diagnostic and therapeutic strategies, as well as epidemiological and conservation studies.


An Extracellular Tetrathionate Hydrolase from the Thermoacidophilic Archaeon Acidianus Ambivalens with an Activity Optimum at pH 1.
The thermoacidophilic and chemolithotrophic archaeon Acidianus ambivalens is routinely grown with sulfur and CO(2)-enriched air. We had described a membrane-bound, tetrathionate (TT) forming thiosulfate:quinone oxidoreductase. Here we describe the first TT hydrolase (TTH) from Archaea. A. ambivalens cells grown aerobically with TT as sole sulfur source showed doubling times of 9 h and final cell densities of up to 8 × 10(8)/ml. TTH activity (≈0.28 U/mg protein) was found in cell-free extracts of TT-grown but not of sulfur-grown cells. Differential fractionation of freshly harvested cells involving a pH shock showed that about 92% of the TTH activity was located in the pseudo-periplasmic fraction associated with the surface layer, while 7.3% and 0.3% were present in the soluble and membrane fractions, respectively. The enzyme was enriched 54-fold from the cytoplasmic fraction and 2.1-fold from the pseudo-periplasmic fraction. The molecular mass of the single subunit was 54 kDa. The optimal activity was at or above 95°C at pH 1. Neither PQQ nor divalent cations had a significant effect on activity. The gene (tth1) was identified following N-terminal sequencing of the protein. Northern hybridization showed that tth1 was transcribed in TT-grown cells in contrast to a second paralogous tth2 gene. The deduced amino acid sequences showed similarity to the TTH from Acidithiobacillus and other proteins from the PQQ dehydrogenase superfamily. It displayed a β-propeller structure when being modeled, however, important residues from the PQQ-binding site were absent. The soluble, extracellular, and acidophilic TTH identified in TT-grown A. ambivalens cells is essential for TT metabolism during growth but not for the downstream processing of the TQO reaction products in S°-grown cells. The liberation of TTH by pH shock from otherwise intact cells strongly supports the pseudo-periplasm hypothesis of the S-layer of Archaea.


Annual Meeting of the International Society of Cancer Metabolism (ISCaM): Metabolic Networks in Cancer.
Cancers are metabolic entities wherein cancer cells adapt their metabolism to their oncogenic agenda and microenvironmental influences. Metabolically different cancer cell subpopulations collaborate to optimize nutrient delivery with respect to immediate bioenergetic and biosynthetic needs. They can also metabolically exploit host cells. These metabolic networks are directly linked with cancer progression, treatment resistance and relapse. Conversely, metabolic alterations in cancer are exploited for anticancer therapy, imaging and personalized medicine. These topics were addressed at the 3rd annual meeting of the International Society of Cancer Metabolism (ISCaM) in Brussels, Belgium, on 26-29 October 2016.


[The effect of riboflavin on lipid peroxidation in rats].
In order to study the effect of riboflavin on lipid peroxidation in rats, randomized block design was used to study the effect of different riboflavin levels on the riboflavin status and lipid peroxidation of S.D. rats. The rats fed with riboflavin deficient diets for 6 weeks had higher levels of BGRAC, blood malondialdehyde and lower levels of GSH than those with riboflavin sufficient diets. The RBC SOD activity is also reduced in riboflavin deficiency rats. It is concluded that riboflavin deficiency increases the extent of lipid peroxidation.


Predictors of children's involvement in parents' treatment among homeless veterans in community residential care.
This study identified the proportion of homeless veterans who are parents and described characteristics of homeless veterans whose children became involved in their treatment. Of the 9,444 veterans surveyed, 37.7 percent were parents of children younger than 18 years; yet children were involved in parents' treatment in only 10.6 percent of these cases. Parents whose children were involved in their treatment were more likely to have direct custody, be female, have greater social stability, and have more psychiatric and medical problems. These parents were also less likely to have been exposed to combat fire. Services to homeless parents might be improved through coordination of adult- and child-focused funding streams and programmatic efforts to provide comprehensive interventions.


Right Atrial Perforation Leading to Cardiac Tamponade Following Veno-Venous Extracorporeal Membrane Oxygenation Cannulation.
Extracorporeal membrane oxygenation (ECMO), as a supportive modality for cardiopulmonary failure, is increasing in its use due to improved advances in technology and experience lending to availability and ease of implementation. Complications with ECMO are quite common, and with increasing use, an increase in complications are a natural result. These complications can be from the underlying disease process or from the ECMO process itself, including cannula insertion. One such complication includes perforation of surrounding structures at site of insertion. We will present a case of right atrial perforation after single lumen cannula insertion, which led to development of cardiac tamponade and subsequently cardiac arrest. In addition to cannula design, lack of wire rigidity can play a role in wire migration and injury to surrounding structures. We emphasize the importance of ultrasound guidance and surveillance with echocardiogram or fluoroscopy during ECMO cannulation, regardless of cannula type, to prevent fatal complications.


Safety of oral direct acting antiviral regimens for chronic hepatitis C in real life conditions.
Direct acting antivirals (DAA) are extremely effective to treat chronic hepatitis C. The aim of this study was to evaluate, by using objective variables, the safety of DAA combinations under clinical practice conditions. A retrospective study was carried out in mono-infected patients with chronic hepatitis C treated with DAA between January and December 2015 in our centre. Discontinuations, treatment modifications, deaths and laboratory parameters were studied (liver function tests, hemoglobin, creatinine and lipid profile at baseline, weeks 4, 8 and post 12). Temporal variation of laboratory parameters was analyzed by t-test for paired data, and comparison between groups was made by t-test for independent samples and ANOVA. 227 patients were included (40.5% cirrhotic). Sustained virological response (SVR) was achieved in 97.3% of patients. In only one case was the antiviral medication suspended due to toxicity, and there were no voluntary treatment discontinuations. The use of ribavirin (RBV) was associated with mild transient hyperbilirubinemia (41.2%) and anemia (32.6%, with RBV dose reduction in 7.9% of cases). There was an elevation in total cholesterol and LDL-cholesterol (LDL-C) during and after treatment: mean increase of 23 mg/dL (0.59 mmol/L) and 22 mg/dL (0.57 mmol/L), respectively in post 12 (p < .0001). An increment of 20% of patients with cholesterol levels over optimal figures was observed after DAA completion. DAA have an optimum safety profile in real life conditions, with infrequent discontinuation and minor laboratory alterations.


Anaphylactic release of thromboxane A2, prostaglandin D2, and prostacyclin from human lung parenchyma.
Antigen challenge of passively sensitized chopped human lung resulted in the generation of several arachidonic acid cyclooxygenase metabolites (AACM): thromboxane A2 (TxA2) as measured by its stable metabolite TxB2, prostaglandin D2 (PgD2), prostacyclin (PgI2) as measured by its stable metabolite 6-keto-PgF1 alpha, prostaglandin F2 alpha (PgF2 alpha), and prostaglandin E (PgE). The kinetics of AACM release after antigen challenge paralleled histamine release. All AACM were released in an antigen dose-dependent manner and reached maximal release at antigen concentrations lower than those required for maximal histamine release. Quantitatively, of the AACM measured, PgD2 and PgI2 were found to predominate in anaphylactic reactions of human lung parenchyma. Generation of PgD2 and PgI2 were 3- to 7-fold greater than that of other AACM measured. Thromboxane B2 was generated in quantities comparable to PgE and PgF2 alpha. Studies were designed to test the hypothesis that lung smooth muscle contraction per se can account for the generated AACM that are released during anaphylaxis of the lung. The studies compared antigen-induced AACM generation with methacholine-induced (10(-4) M) AACM generation. The failure to confirm this hypothesis was especially evident for PgD2 where release was dependent on mast cell activation. Thromboxane A2, PgD2, and PgI2 have been reported to have potent effects on smooth muscle. Our data suggested that these AACM are generated in such sufficient quantities that they may function in important aspects of the modulation of hypersensitivity responses in human lungs.