Impacts of farmland application of antibiotic-contaminated manures on the occurrence of antibiotic residues and antibiotic resistance genes in soil: A meta-analysis study.
A meta-analysis of 94 published studies was conducted to explore the impacts of farmland application of antibiotic-contaminated manures on antibiotic concentrations and ARG abundances in manure-amended soil. Forty-nine antibiotics were reported, in which chlortetracycline, oxytetracycline, doxycycline, tetracycline, enrofloxacin, ciprofloxacin and norfloxacin were the most prevalent and had relatively high concentrations. The responses of ARG and mobile genetic element (MGE) abundances to farmland application of antibiotic-contaminated manures varied considerably under different management strategies and environmental settings. On average, compared to unamended treatments, farmland application of antibiotic-contaminated manures significantly increased the total ARG and MGE abundances by 591% and 351%, respectively (P < 0.05). Of all the included ARG classes, the largest increase was found for sulfonamide resistance genes (1121%), followed by aminoglycoside (852%) and tetracycline (763%) resistance genes. Correlation analysis suggested that soil organic carbon (SOC) was significantly negatively correlated with antibiotic concentrations in manured soil (P < 0.05) due to the formation of covalent bonds and nonextractable residues. Soil silt content was significantly positively correlated with antibiotic concentration (P < 0.05), which was attributed to greater sorption capacities. The ARG abundances were significantly positively correlated with soil silt content, antibiotic concentrations, mean annual temperature, SOC, MGEs and soil pH (P < 0.05), suggesting that changes in these factors may shape the ARG profiles. Collectively, these findings advanced our understanding of the occurrence of antibiotics and ARGs in manure-amended soil and potential factors affecting them and will contribute to better management of these contaminants in future agricultural production.


Cyclic AMP modulation of human B cell proliferative responses: role of cAMP-dependent protein kinases in enhancing B cell responses to phorbol diesters and ionomycin.
The ability of cyclic AMP (cAMP) to modulate human B cell proliferative responses and the possible role of cAMP-dependent kinases (PKA) in cAMP modulation of proliferative responses were investigated. The addition of dibutyl cAMP (Bt2 cAMP) or the cAMP-elevating agent forskolin to B cells stimulated by crosslinking surface immunoglobulins (sIg) resulted in a concentration-dependent inhibition of proliferative responses. By contrast, Bt2 cAMP or forskolin enhanced the proliferative responses of B cells after direct stimulation by phorbol myristate acetate (PMA) and the calcium ionophore ionomycin. The inhibition and enhancement of B cell proliferative responses by Bt2 cAMP were observed at different incubation intervals and were not due to temporal shifts of optimal responses. Also, Bt2 cAMP caused only small changes in B cell RNA synthesis compared to modulation of proliferative responses. Exposure of B cells to Bt2 cAMP rapidly activated PKA. Blocking Bt2 cAMP activation of PKA with the kinase inhibitor HA1004 prevented Bt2 cAMP enhancement of B cell responses after direct stimulation by PMA and ionomycin. In reciprocal experiments, the kinase inhibitor H7 resulted in some inhibition of PKC activation but did not inhibit Bt2 cAMP activation of PKA or Bt2 cAMP enhancement of proliferative responses. Other experiments demonstrated that B cells treated with Bt2 cAMP had selective increases in the de novo phosphorylations of two endogenous substrates which reflected PKA activation. Furthermore, concentrations of HA1004 or H8 which inhibited Bt2 cAMP enhancement of proliferative responses also inhibited PKA phosphorylations of these substrates whereas H7 did not. Thus, elevations of cAMP can enhance or inhibit human B cell proliferative responses to different stimuli and the activation of PKA is important for cAMP enhancement of certain responses.


SARS-CoV-2 Nsp8 induces mitophagy by damaging mitochondria.
Autophagy plays an important role in the interaction between viruses and host cells. SARS-CoV-2 infection can disrupt the autophagy process in target cells. However, the precise molecular mechanism is still unknown. In this study, we discovered that the Nsp8 of SARS-CoV-2 could cause an increasing accumulation of autophagosomes by preventing the fusion of autophagosomes and lysosomes. From further investigation, we found that Nsp8 was present on mitochondria and can damage mitochondria to initiate mitophagy. The results of experiments with immunofluorescence revealed that Nsp8 induced incomplete mitophagy. Moreover, both domains of Nsp8 orchestrated their function during Nsp8-induced mitophagy, in which the N-terminal domain colocalized with mitochondria and the C-terminal domain induced auto/mitophagy. This novel finding expands our understanding of the function of Nsp8 in promoting mitochondrial damage and inducing incomplete mitophagy, which helps us to understand the etiology of COVID-19 as well as open up new pathways for creating SARS-CoV-2 treatment methods.


Hemophagocytic Lymphohistiocytosis Secondary to Ehrlichia Chaffeensis in Adults: A Case Series From Oklahoma.
Hemophagocytic lymphohistiocytosis (HLH) is a syndrome of pathologic immune activation that occurs as either a familial disorder or as a sporadic condition in association with a variety of triggers. Infections are the most common cause of HLH in adults and should be searched for as early treatment usually results in a favorable outcome. Human monocytotropic ehrlichiosis (HME) is a very rare cause of HLH. Failure to consider ehrlichiosis can result in misdiagnosis and an increased length of hospitalization and healthcare cost as described in our report. Treatment for secondary HLH is aimed at reducing hypercytokinemia and eradicating inflammatory and infected cells. It is important to promptly initiate doxycycline when tick-borne diseases are being entertained as a possible trigger, as the antibiotic is effective, safe and inexpensive.


Relationship between appearance of tongue carcinoma on intraoral ultrasonography and histopathologic findings.
To clarify whether intraoral ultrasonography (I-US) is effective for predicting metastasis of tongue cancer to the cervical lymph nodes. Participants comprised 29 patients with tongue carcinoma classified as T1-T4 using the TNM staging system. All patients underwent I-US preoperatively. Postoperatively, resected specimens were evaluated histopathologically. I-US found that cases with invasive depth ≥3 mm had higher potential for cervical lymph node metastasis than those with invasive depth <3 mm (P < .05). No other significant relationships were identified between observations on I-US and cervical lymph node metastasis. Cases with histopathologic blood vessel infiltration or lymph duct infiltration had a significant difference in risk of cervical lymph node metastasis. I-US is useful for preoperatively assessing the invasive depth of tongue carcinoma. Furthermore, observations from I-US and invasive depth of the tumor allowed presumptive diagnosis with regard to cervical lymph node metastasis.


Salicylic acid degradation by advanced oxidation processes. Coupling of solar photoelectro-Fenton and solar heterogeneous photocatalysis.
A 3.0 L solar flow plant with a Pt/air-diffusion (anode/cathode) cell, a solar photoreactor and a photocatalytic photoreactor filled with TiO2-coated glass spheres has been utilized to couple solar photoelectro-Fenton (SPEF) and solar heterogeneous photocatalysis (SPC) for treating a 165mgL(-1) salicylic acid solution of pH 3.0. Organics were destroyed by OH radicals formed on the TiO2 photocatalyst and at the Pt anode during water oxidation and in the bulk from Fenton's reaction between added Fe(2+) and cathodically generated H2O2, along with the photolytic action of sunlight. Poor salicylic acid removal and mineralization were attained using SPC, anodic oxidation with electrogenerated H2O2 (AO-H2O2) and coupled AO-H2O2-SPC. The electro-Fenton process accelerated the substrate decay, but with low mineralization by the formation of byproducts that are hardly destroyed by OH. The mineralization was strongly increased by SPEF due to the photolysis of products by sunlight, being enhanced by coupled SPEF-SPC due to the additional oxidation by OH at the TiO2 surface. The effect of current density on the performance of both processes was examined. The most potent SPEF-SPC process at 150mAcm(-2) yielded 87% mineralization and 13% current efficiency after consuming 6.0AhL(-1). Maleic, fumaric and oxalic acids detected as final carboxylic acids were completely removed by SPEF and SPEF-SPC.


A novel Halomonas ventosae-specific virulent halovirus isolated from the Qiaohou salt mine in Yunnan, Southwest China.
Although Halomonas phages belonging to the families Myoviridae and Siphoviridae have been reported, no virulent Halomonas siphoviruses are known. In this study, a virulent bacteriophage, QHHSV-1, of the family Siphoviridae that specifically infects H. ventosae QH52-2 was isolated from the Qiaohou salt mine. Restriction analysis indicated that QHHSV-1 is a dsDNA virus with a genome size of 33.5-39.5 kb. Transmission electron microscopy showed that QHHSV-1 is a typical representative of the Siphoviridae, with an icosahedral head (47 nm in diameter) and a non-contractile tail (75 nm in length). We also assessed the adsorption rate of QHHSV-1 for the host bacterium and found significant inhibition after the addition of 10 mM CaCl2. Based on a one-step growth curve, we determined a latent period of 30 min and a burst size of 73 PFU/infected cell. At the optimal pH of 8.0, 25.9 and 15.2 % of the phages survived after a 60-min incubation at 50 and 60 °C, respectively. Phage replication was possible at a wide range of salt concentrations, from 2.0 to 20 % (w/v), with an optimum concentration of 5 %. The survival of QHHSV-1 at different salt concentrations decreased with time and 25 % survival after 25 days at 30 % salt concentration.


Forkhead box transcription factor 1: role in the pathogenesis of diabetic cardiomyopathy.
Diabetic cardiomyopathy (DCM) is a disorder of the heart muscle in people with diabetes that can occur independent of hypertension or vascular disease. The underlying mechanism of DCM is incompletely understood. Some transcription factors have been suggested to regulate the gene program intricate in the pathogenesis of diabetes prompted cardiac injury. Forkhead box transcription factor 1 is a pleiotropic transcription factor that plays a pivotal role in a variety of physiological processes. Altered FOXO1 expression and function have been associated with cardiovascular diseases, and the important role of FOXO1 in DCM has begun to attract attention. In this review, we focus on the FOXO1 pathway and its role in various processes that have been related to DCM, such as metabolism, oxidative stress, endothelial dysfunction, inflammation and apoptosis.


Air embolism and the radial arterial line.
A clinical case of apparent air embolism through a radial artery line served as the impetus to conduct a study in primates to investigate the possibility of air embolism from the radial artery into the cerebral circulation. Using 133Xe cerebral scanning, we demonstrated that greater than 2 ml of air in the radial artery would result in retrograde passage of air into the vertebral system and then into the brain.


Gold Nanoclusters Grown on MoS2 Nanosheets by Pulsed Laser Deposition: An Enhanced Hydrogen Evolution Reaction.
Au nanoparticles were decorated on a 2H MoS2 surface to form an Au/MoS2 composite by pulse laser deposition. Improved HER activity of Au/MoS2 is evidenced by a positively shifted overpotential (-77 mV) at a current density of -10 mA cm-2 compared with pure MoS2 nanosheets. Experimental evidence shows that the interface between Au and MoS2 provides more sites to combine protons to form an active H atom. The density functional theory calculations found that new Au active sites on the Au and MoS2 interface with improved conductivity of the whole system are essential for enhancing HER activity of Au/MoS2.


Coordinate regulation of tubulin and microtubule associated protein genes during development of hamster brain.
The present study documents the patterns of mRNA expression for 5 different tubulin genes and 4 of the structural microtubule-associated protein (MAP) genes during normal development of hamster forebrain. Northern blotting in conjunction with densitometric analysis was used to study changes in the levels of the mRNAs for alpha 1-tubulin, classes beta I-, beta II-, beta III- and beta IV-tubulin, as well as the mRNAs for tau, MAP1A, MAP1B and MAP2, using total RNA isolated from hamster forebrain at various embryonic (E) and postnatal (P) stages. Densitometric analyses of the autoradiograms from the Northern blots revealed that each of the tubulin genes exhibited distinct developmental patterns of expression, several of which appeared to be temporally correlated with the expression of specific MAP mRNAs. The beta I-, beta II- and beta III-tubulin mRNAs increased rapidly between late embryonic stages to birth, reached peak levels early in the first postnatal week, and declined thereafter. alpha 1-Tubulin mRNA was easily detected during embryonic stages, rose to peak levels at P7-P9 and then gradually declined. A similar pattern was seen for tau mRNA. After the first postnatal week, the size of the tau mRNA also shifted to a slightly larger size, presumably due to differential splicing. beta IV-Tubulin mRNA levels did not become significant until very late in postnatal development (3-4 weeks). MAP2 mRNA expression was unusual in that peak levels were reached at two different stages of development--an initial peak occurred in the first postnatal week, followed by a decline, and then a second rise occurred during the third and fourth postnatal weeks. The expression of the beta IV-tubulin mRNA coincided temporally with the second peak in MAP2 mRNA expression. MAP1B mRNA abruptly reached high levels at birth, remained abundant during the first two postnatal weeks, and then decreased. In contrast, MAP1A mRNA levels were low in the initial postnatal interval and increased only at very late developmental stages. The findings of a temporal correspondence in expression of high levels of tau and MAP1B with beta I-, beta II-, beta III- and alpha 1-tubulin mRNAs suggest that this profile of gene expression is one that endows greater plasticity to the neuronal cytoskeleton. Conversely, the transition to increased expression of beta IV-tubulin, MAP1A, and a larger tau mRNA species defines a portion of the molecular pattern that underpins the increased stability of neuronal form during maturation.


Harvesting organs for paediatric transplantation: medical features.
The progress in organ transplantation has led to a rise in the demand for organs. Paediatric intensive care units are the main source for obtaining organs. Every "brain dead" patient should be regarded as a potential donor. General contraindications to organ donation are: systemic viral or bacterial infections and extra-cerebral malignancy. They are also organ-specific contraindications. The criteria for the diagnosis of "brain death" have been widely studied and defined. Care of brain dead donors consists of maintenance of cardiac, pulmonary and renal function. Monitoring requires control of central venous and arterial blood pressure, core temperature, urine flow, heart rate and biological data on both urine and blood. A 5% dextrose infusion is maintained with added potassium and sodium chloride according to the urine flow, detectable water loss, and blood and urine composition. Hypotension due to hypovolaemia requires immediate treatment with blood, colloid or albumin infusion. Persistent hypotension with cardiac pump failure is treated with inotropic agents: dopamine and/or dobutamine. Raised urine flow due to diabetes insipidus requires desmopressin (dDAVP). Progress in organ collection requires the consent of the parents and highly motivated medical teams.


The therapeutic potential of neural stem/progenitor cells in murine globoid cell leukodystrophy is conditioned by macrophage/microglia activation.
Twitcher (GALC(twi/twi)) is the murine model of globoid cell leukodystrophy (GLD or Krabbe disease), a disease caused by mutations of the lysosomal enzyme galactocerebrosidase (GALC). To verify the therapeutic potential on twitcher of neural stem/progenitor cells (NSPC), we transduced them with a GALC lentiviral vector. Brain injection of NSPC-GALC increased survival of GALC(twi/twi) from 36.1 +/- 4.1 to 52.2 +/- 5.6 days (P < 0.0001). Detection of GALC activity and flow cytometry showed that NSPC-GALC and NSPC expressing the green fluorescent protein were attracted to the posterior area of twitcher brain, where demyelination occurs first. GALC(twi/twi) microglia, also more abundant in posterior regions of the brain, released significant amounts of the cytotoxic cytokine TNF-alpha when matched with NSPC-GALC. Thus, in murine GLD, and possibly in other demyelinating diseases, NSPC are attracted to regions of active demyelination but have limited survival and therapeutic potential if attacked by activated macrophages/microglia.


Rationale and design of PASSAT - patients' satisfaction with local or general anaesthesia in video-assisted thoracoscopic surgery: study protocol for a randomised controlled trial with a non-randomised side arm.
Although general anaesthesia (GA) with one-lung ventilation is the current standard of care, minor thoracoscopic surgery, i.e. treatment of pleural effusions, biopsies and small peripheral pulmonary wedge resections, can also be performed using local anaesthesia (LA), analgosedation and spontaneous breathing. Whilst the feasibility and safety of LA have been demonstrated, its impact on patient satisfaction remains unclear. Most studies evaluating patient satisfaction lack control groups or do not meet psychometric criteria. We report the design of the PASSAT trial (PAtientS' SATisfaction in thoracic surgery - general vs. local anaesthesia), a randomised controlled trial with a non-randomised side arm. Patients presenting for minor thoracoscopic surgery and physical eligibility for GA and LA are randomised to surgery under GA (control group) or LA (intervention group). Those who refuse to be randomised are asked to attend the study on the basis of their own choice of anaesthesia (preference arm) and will be analysed separately. The primary endpoint is patient satisfaction according to a psychometrically validated questionnaire; secondary endpoints are complication rates, capnometry, actual costs and cost effectiveness. The study ends after inclusion of 54 patients in each of the two randomised study groups. The PASSAT study is the first randomised controlled trial to systematically assess patients' satisfaction depending on LA or GA. The study follows an interdisciplinary approach, and its results may also be applicable to other surgical disciplines. It is also the first cost study based on randomised samples. Comparison of the randomised and the non-randomised groups may contribute to satisfaction research. German Clinical Trials Register, DRKS00013661 . Registered on 23 March 2018.


Transection of the retrohepatic vena cava in a case of total situs inversus following blunt trauma.
Total transection of the retrohepatic inferior vena cava from blunt trauma is a rare and usually fatal injury. Such an injury in a patient with total situs inversus and previous surgical correction of great vessel transposition is described. A possible mechanism for the injury is suggested.


Guiding Cell Network Assembly using Shape-Morphing Hydrogels.
Forces and relative movement between cells and extracellular matrix (ECM) are crucial to the self-organization of tissues during development. However, the spatial range over which these dynamics can be controlled in engineering approaches is limited, impeding progress toward the construction of large, structurally mature tissues. Herein, shape-morphing materials called "kinomorphs" that rationally control the shape and size of multicellular networks are described. Kinomorphs are sheets of ECM that change their shape, size, and density depending on patterns of cell contractility within them. It is shown that these changes can manipulate structure-forming behaviors of epithelial cells in many spatial locations at once. Kinomorphs are built using a new photolithographic technology to pattern single cells into ECM sheets that are >10× larger than previously described. These patterns are designed to partially mimic the branch geometry of the embryonic kidney epithelial network. Origami-inspired simulations are then used to predict changes in kinomorph shapes. Last, kinomorph dynamics are shown to provide a centimeter-scale program that sets specific spatial locations in which ≈50 µm-diameter epithelial tubules form by cell coalescence and structural maturation. The kinomorphs may significantly advance organ-scale tissue construction by extending the spatial range of cell self-organization in emerging model systems such as organoids.


Secretion of rabbit C-reactive protein by transfected human cell lines is more rapid than by cultured rabbit hepatocytes.
C-reactive protein (CRP) is a major acute phase protein in humans and rabbits. Its synthesis by the liver varies over a 1000-fold range depending on the presence and severity of inflammatory stimuli. In previous studies of synthesis and secretion of rabbit CRP, we showed that secretion becomes more efficient over the course of the acute phase response as CRP synthesis rates increase (Macintyre, S.S., Kushner, I., and Samols, D. (1985) J. Biol. Chem. 260, 4169-4173). The current studies were undertaken to help distinguish between two alternative explanations for this finding: 1) that secretion efficiency may simply be a property of the rate of synthesis and intracellular concentration of CRP or 2) that secretion may be regulated by separate intracellular mechanisms. A fusion gene containing the mouse metallothionein I promoter linked to the protein coding region of the rabbit CRP gene was introduced into the human hepatoma cell line, NPLC, and the nonliver cell line, HeLa. In this system a graded response of the mouse metallothionein I promoter following exposure to increasing zinc concentrations results in increasing CRP synthesis. Unlike hepatocytes from rabbits undergoing the acute phase response, we found that rabbit CRP was secreted by these transfected cell lines with a very high degree of efficiency which was independent of the rate of CRP synthesis. This finding implies that normal rabbit hepatocytes retard the secretion of CRP and that this inhibition is diminished as the acute phase response progresses. It further indicates that the relationship between changes in synthetic rate and efficiency of secretion of rabbit CRP is not a causal one and that synthesis and secretion of CRP by rabbit hepatocytes are regulated by independent intracellular mechanisms during the acute phase response.


Spirometry and flow-volume curves in healthy, normal Pakistanis.
Previous studies have indicated that lung volumes in healthy, normal Pakistani adults are smaller than measurements reported in comparable healthy European populations; in order to confirm these findings and to examine the relationship of maximal expiratory flow rates to lung volumes, we studied 250 non-smoking healthy subjects (116 men and 114 women) between the ages of 18 and 65 years. The population sample was drawn from urban and rural areas of Pakistan, with low levels of air pollution. The results indicate that the forced vital capacity (FVC) and forced expired volume in 1 second (FEV1) were lower in the Pakistani population compared to European populations and North American populations of European descent. These data are in conformity with previous studies; however, in Pakistani men the effects of age on FVC and FEV1 were slight so that, after the fourth decade, the FVC and FEV1 values are very comparable between the European and Pakistani populations. Amongst Pakistani women, on the other hand, FVC and FEV1 remained lower than in their European counterparts throughout adult life. Maximal expiratory flow rates amongst the men did not correlate with age, and these values were very similar to those reported in age-matched European populations. In women, however, there was a significant correlation of maximal flow rates with age and height, and the maximal expiratory flows were decreased compared to European populations. These data indicate that in Pakistani men pulmonary mechanics may be different to their European counterparts, allowing for higher maximal expiratory flows at any given lung volume.


Evaluation of fixation stability using different targets with the MP1 microperimeter.
The aim of this study was to evaluate fixation stability using two different fixation targets with the Nidek MP1 microperimeter. Twenty-nine healthy subjects with a mean age of 26.53 ± 7.35 years and visual acuity ≥0.0 logMAR were enrolled in this study. Fifty-eight eyes of 29 patients without ophthalmic and/or systemic disease underwent a fixation test with the MP1 microperimeter. Fixation stability related to a red cross (central) and/or a red circle (pericentral) target was quantified using either Fujii classification or by calculating the bivariate contour ellipse area (BCEA). For statistical analysis, BCEA values were converted into their logarithms (logBCEA) and all data obtained were analyzed using paired Student's t test. The inclination values of the axis of BCEA were analyzed with Chi squared test. The mean values of logBCEA and the mean values of the major and minor axis of the ellipses related to the cross and the circle fixation target were significantly different (68.2 %, p = 0.00; 95.4 %, p = 0.00; 99.6 %, p = 0.00, respectively) for each BCEA standard deviation. Fixation was significantly less stable for the pericentral fixation target in normal subjects, indicating an advantage for central fixation targets. These results are of particular interest when evaluation of changes in fixation is needed.


Survival After Mediastinal Node Dissection, Systematic Sampling, or Neither for Early Stage NSCLC.
The American College of Surgeons Oncology Group Z0030 found no survival difference between patients with early stage NSCLC who had mediastinal nodal dissection or systematic sampling. However, a meta-analysis of 1980 patients in five randomized controlled trials from 1989 to 2007 associated better survival with nodal dissection. We tested the survival impact of the extent of nodal dissection in curative-intent resections for early stage NSCLC in a population-based observational cohort. Resections for clinical T1 or T2, N0 or nonhilar N1, M0 NSCLC in four contiguous United States Hospital Referral Regions from 2009 to 2019 were categorized into mediastinal nodal dissection, systematic sampling, and "neither" on the basis of of the evaluation of lymph node stations. We compared demographic and clinical characteristics, perioperative complication rates, and survival after assessing statistical interactions and confounding. Of the 1942 eligible patients, 18% had nodal dissection, 6% had systematic sampling, and 75% had an intraoperative nodal evaluation that met neither standard. In teaching hospitals, nodal dissection was associated with a lower hazard of death than "neither" resections (0.57 [95% confidence interval: 0.41-0.79]) but not systematic sampling (0.74 [0.40-1.37]) after adjusting for multiple comparisons. There was no significant difference in hazard ratios at nonteaching institutions (p > 0.3 for all comparisons). Perioperative complication rates were not significantly worse after mediastinal nodal dissection or systematic sampling, compared with "neither," (p > 0.1 for all comparisons). In teaching institutions, mediastinal nodal dissection was associated with superior survival over less-comprehensive pathologic nodal staging. There was no survival difference between teaching and nonteaching institutions, a finding that warrants further investigation.


Is Video-Based Analysis a Valid Method for Determining Mechanisms of Ankle Injuries During Gameplay in the National Basketball Association?
In the National Basketball Association (NBA), lower extremity injuries account for over 70% of games missed, with ankle injuries being the most common. High-quality video analysis has been successful for studying injury mechanism. To (1) determine the validity of video-based analysis as a method to evaluate ankle injury mechanisms in NBA players and (2) analyze the circumstances associated with injury, games missed due to injury, and associated costs in player salary due to time missed. Case series; Level of evidence, 4. Ankle injuries were identified using an injury report database, and corresponding videos were searched using YouTube.com to access high-quality video evidence of these injuries during the 2015-2020 NBA regular season. We reviewed 822 injuries, of which 93 had corresponding videos (video subset), in our final analysis. Variables including number of games missed, necessity for surgical treatment, and injury recurrence were reported for the entire cohort. In the video subset, the mechanism of injury and other corresponding situational data were evaluated. The most common mechanism of injury occurred via ankle inversion (83.9%; n = 78; P < .001). These injuries were significantly associated with indirect contact with the player's ankle (79.6%; n = 74; P < .001). There were significant differences based on player position, within both the video subset (P = .008) and the entire cohort (P < .001), with guards being injured the most frequently. The average number of games missed due to injury was 7 games in the video subset and 5 games in the entire cohort (P = .14). There were significant differences between the groups in average player salary per game ($133,878 [video subset] versus $87,577 [entire cohort]; P < .001). Despite its low yield of 11.3%, video analysis proved to be a useful tool to determine ankle injury mechanisms as well as the distribution of injuries based on player position. However, this methodology was subject to selection bias, as evidenced by a $50,000 increase in player salary among the video cohort. These findings should be considered when using video analysis in future studies.


Efficient metal-free synthesis of various pyrido[2',1':2,3]imidazo- [4,5-b]quinolines.
Dancing with diversity: The synthesis of diverse pyrido[2',1':2,3]imidazo[4,5-b]quinolines bearing several substitution patterns was developed based on combining a multicomponent reaction (Groebke-Blackburn-Bienaymé reaction) with an original cyclization as a secondary transformation (see scheme; DBU = 1,8-diazabicyclo[5.4.0]undec-7-ene).


Synthesis of oligosaccharide fragments of the repeating unit of Salmonella kentucky O-specific polysaccharide and conversion of the oligosaccharides into the glycosyl phosphates.
alpha-D-Man-(1----2)-alpha-D-Man-(1----3)-D-Gal, a structural fragment of the main chain of Salmonella serogroups C2 and C3 O-specific polysaccharides, and the isomer with the central residue beta have been synthesised, as have some oligosaccharides related to the structure of the O-specific polysaccharide of S. kentucky (serogroup C3), namely, alpha-D-Glc-(1----4)-D-Gal, alpha-D-Man-(1----3)-[alpha-D-Glc-(1----4)]-D-Gal, and alpha-D-Man-(1----2)-alpha-D-Man-(1----3)-[alpha-D-Glc-(1----4)]-D-Gal, and the isomers with the D-Glc unit beta. Each oligosaccharide was converted into the alpha-glycosyl phosphate.


'It will dictate how many children I will have"- Women's decision-making in pregnancy following a previous caesarean birth (CB).
Vaginal birth after caesarean (VBAC) is supported in systematic reviews (Wu et al., 2019) and national guidelines (RCOG, 2015) and women are expected to be involved in the decision-making process for either a repeat caesarean birth or planned VBAC. To develop a Grounded Theory (GT) of women's decision making of their birth choices in pregnancy following a previous caesarean birth (CB) OBJECTIVE: To explore what determines women's birth choice and their decision making for birth following a previous CB. Semi structured interviews with pregnant women were undertaken in order to develop a Glasserian Grounded Theory SETTING: Antenatal clinics and wards in a large tertiary level maternity hospital. The theory of 'Mentalizing Possibilities' is a substantive theory which explains pregnant women's decision making about their birth choices after a previous CB. Women's main concern is to achieve a positive experience. The core category of 'Mentalizing Possibilities' explains how women process their previous experience, adapt to uncertainty and deal with the decisional conflict. There are behavioural and cognitive strategies which women use to go through this process. Women want a positive birth experience after a previous CB and require support and continuity in decision making to help them decide the optimal birth choice for their current pregnancy.


Accelerating cancer clinical trial recruitment through a financial reimbursement program integrated with patient navigation: an interrupted time series analysis.
Cancer treatment clinical trials face major challenges with patient recruitment. Strategies to address patient indirect costs associated with clinical trial participation may accelerate accrual overall. The current study examined the effect of the IMproving Patient Access to Clinical Trials (IMPACT) intervention on patient accrual to cancer treatment clinical trials at an academic medical center. The IMPACT intervention was an onsite patient navigator combined with a financial reimbursement program to address patient out of pocket costs and began on September 2018. This analysis measured frequency of patient enrollment in cancer treatment clinical trials and available cancer treatment clinical trials per month between January 1, 2016 and March 31, 2020. An interrupted time-series analysis (ITSA) was conducted to estimate changes in patient enrollment attributable to the IMPACT intervention. During the study period, a mean of 69 patients enrolled in clinical trials per month (standard deviation (SD = 13), with 27 (SD = 7) in early phase vs 41 (SD = 12) in late phase clinical trials. The number of available clinical trials per month was 51 (SD = 2) overall, with 23 (SD = 1) in early phase vs 28 (SD = 1) in late phase context. A total of 3470 patients were enrolled in cancer treatment clinical trials during the evaluated time period, the majority of whom were men (1895, 55 %) and racially white (2267, 65 %). A statistically significant increase in the number of patients accrued as compared to the pre-intervention trend was observed; with approximately 1 additional patient accrued per month, with a larger effect on increase patient accrual for late phase clinical trials. This study observed that the IMPACT intervention accelerated clinical trial recruitment, especially among late phase clinical trials. Future research will examine strategies to leverage this infrastructure to optimize recruitment among underrepresented patients. To improve clinical trial recruitment and ensure that trial results are representative of a diverse population it is critical for health policies consider patient out-of-pocket costs and potential reimbursement to alleviate financial burden associated with clinical trial participation. Furthermore, policies for facilitating clinical trial recruitment and participant retention should budget for and incorporate a navigation component to assist patients who may not be familiar with the healthcare system and available financial assistance.


Differential time course of antidepressant effects after sleep deprivation, ECT, and carbamazepine: clinical and theoretical implications.
The pattern and time course of antidepressant response to different treatment modalities provide important clinical information and hints about underlying neurobiological mechanisms. Depressed patients who responded to 1 night's sleep deprivation (11 of 33 patients) showed maximal improvement on day 1 and deterioration in mood thereafter. In contrast, slower onset and more sustained effects were observed following carbamazepine (12 of 37) or electroconvulsive therapy (ECT) (8 of 8). Nearly maximal improvement required about 2 weeks for ECT and 3 weeks for carbamazepine. Possible differential or common biological mechanisms with differential times of action are implied by these data, which are of importance to the neuroscientist attempting to uncover neural substrates of antidepressant response and the clinician attempting to find rapid onset, yet sustained antidepressant treatments.


The autonomic dysfunction in patients with ankylosing spondylitis: a clinical and electrophysiological study.
The aim of this study was to determine autonomic nervous system (ANS) functions by using clinical and electrophysiological tests in patients with ankylosing spondylitis (AS). Twenty AS and 20 healthy control subjects were recruited. Demographic data, symptoms related with ANS, and neurological findings were recorded. Clinical measurements including the heart rate variation with deep breathing (HRV), heart rate response to standing (HRS), systolic blood pressure response to standing, and diastolic blood pressure response to isometric exercise were obtained to assess parasympathetic and sympathetic functions of the subjects. The electrophysiological assessments of ANS were performed by sympathetic skin response (SSR) and R-R interval variation (RRIV) measurements for the sympathetic and parasympathetic functions, respectively. Patients with AS were subdivided into two groups depending on the activity of disease. The difference between the groups and relationship between ANS variables and clinical entities were determined. Fifteen male and five female AS patients with a mean age of 38+/-8.05 years and 14 male and six female healthy control subjects with a mean age of 40+/-9.8 years were included in the study. All the subjects were totally symptom free for ANS involvement and had normal neurological examination findings. The levels of HRV, HRS, and the mean RRIV values were significantly lower in AS patients than in control subjects. The clinical ANS parameters of the patients having more active disease were lower than in subjects with mild disease in regard to HRV values and SSR amplitudes and higher in regard to SSR latencies. The HRV values were found to be correlated with the mean scores of Bath ankylosing spondylitis disease activity index (BASDAI) and C-reactive protein (CRP) levels, and the mean latencies of SSR were correlated with BASDAI scores and CRP levels. In conclusion, our study indicates a subclinical mainly parasympathetic dysfunction of ANS in patients with AS which can be related with disease activity.


Overview, Generation, and Significance of Variable New Antigen Receptors (VNARs) as a Platform for Drug and Diagnostic Development.
The approval of the first VHH-based drug caplacizumab (anti-von Willebrand factor) has validated a two-decade long commitment in time and research effort to realize the clinical potential of single-domain antibodies. The variable domain (VNAR) of the immunoglobulin new antigen receptor (IgNAR) found in sharks provides an alternative small binding domain to conventional monoclonal antibodies and their fragments and heavy-chain antibody-derived VHHs. Evolutionarily distinct from mammalian antibody variable domains, VNARs have enhanced thermostability and unusual convex paratopes. This predisposition to bind cryptic and recessed epitopes has facilitated both the targeting of new antigens and new (neutralizing) epitopes on existing antigens. Together these unique properties position the VNAR platform as an alternative non-antibody binding domain for therapeutic drug, diagnostic and reagent development. In this introductory chapter, we highlight recent VNAR advancements that further underline the exciting potential of this discovery platform.


Association of weight change following smoking cessation with the risk of tuberculosis development: A nationwide population-based cohort study.
Smoking or weight loss is a risk of tuberculosis (TB) development. However, the impact of weight change after smoking cessation on the occurrence of TB remains elusive. We aimed to determine the relationship between weight change after smoking cessation and the risk of TB development. We conducted a population-based cohort study using the national database in Republic of Korea. Of the 10,490,491 subjects who underwent health check-up in 2009, we enrolled 9,953,124 subjects without a previous TB history and followed them until 2017. We divided all study participants into the following three groups: never, former, and current smokers. The primary endpoint was newly developed TB. Among 9,953,124 subjects analyzed, 5,922,845 (59.5%) were never smokers, 1,428,209 (14.4%) were former smokers, and 2,602,080 (26.1%) were current smokers. The risk of TB development was significantly higher in current smokers than in never smokers (adjusted hazard ratio (aHR) 1.158; 95% confidence interval [CI] 1.131-1.186). Among current smokers, individuals who stopped smoking and maintained weight after baseline evaluation had a significantly lower risk of TB development compared with those who continued to smoke (aHR 0.771; 95% CI 0.741-0.892). However, even after smoking cessation, individuals who lost weight were at a significantly higher risk of TB development compared with those who continued to smoke (aHR 1.327; 95% CI 1.119-1.715). Our findings suggest that smoking is a risk factor for TB and weight maintenance (neither gaining or losing) after quitting smoking might reduce the risk of TB development.


Clinical pharmacokinetics of aztreonam.
Aztreonam (azthreonam) is practically completely absorbed after intramuscular injection. After intravenous injection plasma concentrations follow a 2-compartment open model, with a t1/2 alpha of 0.20 hours. Volume of distribution at steady-state (Vdss) after intravenous or intramuscular injection is about 0.16 L/kg (0.42 L/kg for the free drug). After oral administration less than 1% of the drug is absorbed. Over a large dosage range plasma concentrations increase linearly with dose. No accumulation occurs after multiple dosing. Plasma binding in healthy subjects is about 56% and is not concentration dependent. Diffusion into tissues is generally slow, and the ratio between mean tissue and plasma concentration seems to depend mainly on the composition of the tissue. In inflamed meninges, penetration of aztreonam into CSF is more rapid than with uninflamed meninges. Diffusion through the placenta is poor, as is diffusion into breast milk. The main route of elimination of aztreonam is by the kidney, partly by active tubular excretion, but this can be inhibited by probenecid. Extrarenal clearance is probably due to excretion by the liver. Metabolism occurs to a very limited extent. Total plasma clearance in healthy adults is about 140 ml/min (8.4 L/h) or 2 ml/min/kg (0.12 L/h/kg), and terminal half-life is 1.7 hours. In children clearance is similar to that in adults when expressed as a function of bodyweight, but in neonates, especially in low birthweight infants, it is less [about 1 ml/min/kg (0.06 L/h/kg)]. In various disease states the Vdss of aztreonam is not appreciably different from that found in healthy individuals.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantitative X-ray microanalysis of diffusible ions in the skeletal muscle bulk specimen.
X-ray microanalysis in the scanning electron microscope (SEM) is used to obtain information about the distribution and mobility of electrolyte ions in the skeletal muscle tissue. In order to localize the diffusible ions in their physiological state, so far as possible, any chemical fixative is avoided. A special 'cooling chain' preparation method is applied which enables studies on cryofractured bulk specimens at low temperatures. Quantitation of the obtained X-ray spectra is done by the use of glycerol-treated tissue specimens which are incubated with electrolyte solutions of defined ion concentration.


Hydrogen-bonded co-crystal structure of benzoic acid and zwitterionic l-proline.
The title compound [systematic name: benzoic acid-pyrrolidin-1-ium-2-carboxyl-ate (1/1)], C7H6O2·C5H9NO2, is an example of the application of non-centrosymmetric co-crystallization for the growth of a crystal containing a typically centrosymmetric component in a chiral space group. It co-crystallizes in the space group P212121 and contains benzoic acid and l-proline in equal proportions. The crystal structure exhibits chains of l-proline zwitterions capped by benzoic acid mol-ecules which form a C(5)[R33(11)] hydrogen-bonded network along [100]. The crystal structure is examined and compared to that of a similar co-crystal containing l-proline zwitterions and 4-amino-benzoic acid.


Sparsity-constrained three-dimensional image reconstruction for C-arm angiography.
X-ray C-arm is an important imaging tool in interventional radiology, road-mapping and radiation therapy because it provides accurate descriptions of vascular anatomy and therapeutic end point. In common interventional radiology, the C-arm scanner produces a set of two-dimensional (2D) X-ray projection data obtained with a detector by rotating the scanner gantry around the patient. Unlike conventional fluoroscopic imaging, three-dimensional (3D) C-arm computed tomography (CT) provides more accurate cross-sectional images, which are helpful for therapy planning, guidance and evaluation in interventional radiology. However, 3D vascular imaging using the conventional C-arm fluoroscopy encounters some geometry challenges. Inspired by the theory of compressed sensing, we developed an image reconstruction algorithm for conventional angiography C-arm scanners. The main challenge in this image reconstruction problem is the projection data limitations. We consider a small number of views acquired from a short rotation orbit with offset scan geometry. The proposed method, called sparsity-constrained angiography (SCAN), is developed using the alternating direction method of multipliers, and the results obtained from simulated and real data are encouraging. SCAN algorithm provides a framework to generate 3D vascular images using the conventional C-arm scanners in lower cost than conventional 3D imaging scanners.


Perfluorooctane sulfonate impairs rat Leydig cell development during puberty.
Perfluorooctane sulfonate (PFOS) possibly delays male sexual development. However, its effects on pubertal Leydig cell development are unclear. The objective of the present study was to investigate the effects of in vivo PFOS exposure on rat Leydig cell development during puberty. Immature male Sprague Dawley rats were gavaged 5 or 10 mg/kg PFOS on postnatal day 35 for 21 days. Compared to the control (0 mg/kg), PFOS lowered serum testosterone levels without altering luteinizing hormone and follicle-stimulating hormone levels on postnatal day 56. PFOS in vivo downregulated mRNA or protein levels of Leydig cells (Lhcgr, Cyp11a1, and Cyp17a1). PFOS in vitro inhibited androgen secretion in immature Leydig cells at ≥ 50 nM, most possibly via downregulating Hsd17b3 mRNA level. At ≥ 500 nM, PFOS downregulated Lhcgr, inhibited BCL-2 and increased BAX levels to cause Leydig cell apoptosis. In conclusion, PFOS at a lower dose directly inhibited pubertal development of Leydig cells.


Revision of the requirements for live vaccine processing; confirmation of effective date. Direct final rule; confirmation of effective date.
The Food and Drug Administration (FDA) is confirming the effective date of March 18, 2008, for the direct final rule that appeared in the Federal Register of October 18, 2007 (72 FR 59000). The direct final rule amends the biologics regulations by providing options to the existing requirements for the processing of live vaccines. This document confirms the effective date of the direct final rule.


Evaluation of pooled cultures for bacterial detection in whole blood-derived platelets.
The BacT/ALERT (bioMérieux) system is highly efficient for bacterial detection in apheresis platelets (PLTs) and whole blood-derived PLTs produced by the buffy-coat method. Detection of bacterial contamination in whole blood-derived PLTs produced by the PLT-rich plasma (PRP) method, however, is problematic. Prestorage pooling of these PLTs is not permitted in some countries including Canada and the United States, and culturing individual units is costly and may significantly reduce the PLT unit content. In this study, the sensitivity and specificity of BacT/ALERT cultures performed on pools derived from PRP PLTs are reported. The sensitivity of the BacT/ALERT system was evaluated for bacterial detection in PRP PLTs with a dilution effect. Thirty PLT pools were produced with 1 PLT unit previously spiked with bacteria and then pooled with other four nonspiked PLT units. Three bacteria, usually associated with PLT contamination, were selected for spiking. The specificity of this method was evaluated in 40 nonspiked PLT pools. The method was found to be 100 percent specific and 97 percent sensitive. Of the five spiked pools with Streptococcus pneumoniae at levels of less than 2 colony-forming units (CFUs) per mL, four were found to be positive whereas all 25 spiked pools with greater than 9 CFUs per mL of any of the chosen bacteria gave positive results. The mean time of detection was 17 to 19 hours for Staphylococcus epidermidis and 14 to 15 hours for S. pneumoniae and Pseudomonas aeruginosa when spiked with similar bacterial inocula. The evaluated system is highly sensitive and specific and may be a feasible method for bacterial detection in PRP PLTs.


[French translation and linguistic validation of the questionnaire Bladder Cancer Index (BCI)].
Translation and linguistic validation of the French version of Bladder Cancer Index (BCI). A double-back translation of the original Bladder Cancer Index was performed. First, two urologists translated the English version in French. Then, a first consensus meeting between the translators and a group composed of urologists and nurses was achieved. Back-translation of this version was then done by professional translators (Nagpal, Paris) to ensure that no distortion was detected between the two questionnaires. Finally, a pilot study followed by an interview was carried out among one woman and five men having bladder cancer. The consensus version is attached to the article. No difficulties were reported by the pilot population to comprehend or to complete this BCI French version. This BCI French version-attached to the article-makes it possible for researchers among a French population to use this validated and internationally recognized tool among a French population. The impact of various bladder cancer treatment on quality of life could hence be assessed and compared.


Usefulness of Abductive Reasoning in Nursing Education: A Pilot Study.
Hypothetico-deductive reasoning used by novice nurses could limit their ability to explain a presenting care situation in its entirety. Hence, scholars recommend the use of abductive reasoning as an alternative approach. This study explored the effects of abductive reasoning training on baccalaureate nursing students' hypothesis generation abilities. Through a pretest-posttest study, we delivered educational training on abductive reasoning and examined hypothesis accuracy, expertise, and breadth. Participants generated scenario-specific hypotheses before and after the training. Academic content experts validated the scenarios, and 2 independent raters scored participants' hypotheses. Twenty first- and second-year nursing students participated in this pilot study. Posttest scores showed a significant improvement in participants' hypothesis generation abilities: accuracy (P < .001), expertise (P < .001), and breadth (P = .006). Abductive reasoning training in nursing education may improve students' hypothesis generation abilities.


Screening for Factors Involved in X Chromosome Inactivation Using Haploid ESCs.
Forward genetics can provide insight into molecular pathways as has been demonstrated by advances in cell biology from comprehensive genetic studies in simple organisms. Recently, techniques have become available that promise a similar potential for understanding developmental pathways in mammals. Here we describe a genetic forward screening approach for identifying factors involved in X chromosome inactivation that is based on haploid mouse embryonic stem cells. Using a genetically encoded selection system screening of large mutant pools can identify candidate silencing factors with high confidence.


Early type I collagen deposition is associated with prognosis in biliary atresia.
Biliary atresia (BA) is a cholestatic liver disease of children that progresses to hepatic fibrosis. BA is the main indication of pediatric liver transplantation (LTx). Histopathological markers in liver biopsies could be useful for predicting progression to end-stage disease. To establish histopathological or immunohistochemical markers in liver biopsies of BA patients and correlate those markers with prognosis. Histological analysis of biliary alterations and morphometric assessment of liver fibrosis were performed, in addition to indirect immunofluorescence assays (IF) for type I, III, IV and V collagens in initial and final liver biopsies of 36 patients with BA who underwent Kasai hepatoportoenterostomy (KPE) and LTx in the last 20years at a single center. Histopathological markers had no correlation with evolutive time until LTx. The perisinusoidal deposition of type III and V collagens was more prominent in the initial biopsies (p<0.01), whereas deposition of type I and IV collagens indicated progression (p<0.01). Patients with large amounts of perisinusoidal type I collagen in the initial biopsies had worse progression time curves until LTx (p=0.04). Morphometric assessment of perisinusoidal deposition of type I collagen by IF in the initial biopsy can correlate with progression time to LTx in post-surgical BA.


Reference LC-MS/MS method to detect fresh cheeses adulteration with whey.
This study evaluated the potential of a reference method to detect fresh cheeses adulteration with whey by liquid chromatography coupled to mass spectrometry (LC-MS/MS). Qualitative results were expressed as presence or absence of the marker peptide TPEVDDEALEK, obtained by tryptic hydrolysis of β-lactoglobulin. Sample preparation was based on defatting with cold acetone and protein solubilization in ammonium bicarbonate and urea buffer (pH = 8.0). Reversed phase liquid chromatography used a C18 column for separation of the analyte, whose retention time was 4.12 min. Mass spectrometry was carried out with positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode for the precursor ion (m/z 624) and the quantitation (m/z 573) and confirmation transitions (m/z 820; m/z 920) of the peptide. Method validation was carried out in quantitative terms, to set the baseline concentration of the marker peptide in 69 authentic samples, and in qualitative terms, to set the action level that distinguish authentic from adulterated cheeses (350 mg kg-1). Sensitivity was enough to detect cheeses with 10% adulteration and the detection limit was set to 21 mg kg-1. The simple extraction procedure allowed high-throughput analysis of 33 real samples. Results were compared to SDS-PAGE electrophoresis, whose limitations for accurate quantitation were easily overcome by LC-MS/MS. The developed method ensured precision, accuracy, sensitivity, and specificity needed for the unequivocal detection of non-compliant cheeses made with cow or buffalo milk, without dealing with the highly toxic chemical species required for SDS-PAGE. This method can be extended in the future to detect similar adulterations in fresh cheeses prepared with milk from other animal species, as well as in other dairy products.


Monoamine metabolites in the CSF of conscious unrestrained cats.
The dopamine metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) and the serotonin metabolite 5-hydroxyindoleacetic acid (5-HIAA) were measured repeatedly over a period of up to 5 months at different sites of the brain ventricular system in unrestrained, awake cats. Samples of 10 microliter CSF were analyzed by high pressure liquid chromatography and subsequent electrochemical detection. Concentrations were in the range of 30-130 ng/ml for DOPAC, 110-340 ng/ml for 5-HIAA and 180-750 ng/ml for HVA. The monoamine metabolites were constant even over a period of several months if measured in the same animal but there was a marked interindividual variation. A marked gradient for monoamine metabolites was found when CSF samples from frontal sites of the lateral ventricle were compared to CSF samples from the dorsal lateral ventricle. The concentrations of DOPAC and HVA were higher at frontal sites.


Traumatic brain injury causes a decrease in M2 muscarinic cholinergic receptor binding in the rat brain.
Numerous studies indicate that an acute, excessive activation of muscarinic acetylcholine receptors (mAChR) contributes to the pathophysiological sequela of TBI. The present study examined the effect of moderate fluid percussion traumatic brain injury (TBI) on binding to M1 and M2 mAChR subtypes in the hippocampal formation and adjacent cortex using quantitative autoradiography. Injured animals along with concurrent controls were sacrificed by in situ freezing at 3 h or 24 h following TBI. Slide-mounted tissue sections were incubated in either [3H]pirenzepine (23 nM) for M1 or [3H]AFDX384 (9 nM) for M2 mAChR subtype labeling. Binding of [3H]pirenzepine to the M1 mAChR subtype was not significantly altered by TBI when compared to sham-injured animals. [3H]AFDX384 binding to the M2 mAChR subtype was significantly decreased at 24 h in hippocampal CA2-3 region and dorsal blade of the dentate gyrus (P < 0.05). The differences observed between M1 and M2 subtypes suggests that these muscarinic subtypes may differentially contribute to the pathophysiology of TBI.


Hypothyroidism induces expression of the peptide transporter PEPT2.
The kidney is a target organ for thyroid hormone action and a variety of renal transport processes are altered in response to impaired thyroid functions. To investigate the effect of thyroid hormone on the expression of the renal proximal tubular high-affinity-type H(+)-peptide cotransporter (PEPT2) in rats, hypothyroidism was induced in animals by administration of methimazole (0.05%) via drinking water. After 7 weeks of treatment, hypothyroidism was confirmed by determining serum free T(3) and free T(4) concentrations. Northern blotting was used to examine the expression of PEPT2 mRNA in kidney tissues from hypothyroid rats compared to control rats. Hypothyroidism resulted in an increased level of total renal PEPT2 mRNA (121.1+/-3.3% vs. control 100+/-2.8%; p=0.008). The mRNA results were confirmed by immuno-blotting, which demonstrated significantly increased protein levels (162% vs. control 100%; p<0.01). Immunohistochemistry also revealed increased PEPT2 protein levels in the proximal tubules of treated compared to non-treated rats. In summary, PEPT2 is the first proximal tubule transporter protein that shows increased expression in states of hypothyreosis. As PEPT2 reabsorbs filtered di- and tripeptides and peptide-like drugs, the present findings may have important implications in nutritional amino acid homeostasis and for drug dynamics in states of altered thyroid function.


Validation of a Sequential Organ Failure Assessment Score using Electronic Health Record Data.
The sequential organ failure assessment (SOFA) score is a scoring system commonly used in critical care to assess severity of illness. Automated calculation of the SOFA score using existing electronic health record data would broaden its applicability. We performed a manual validation of an automated SOFA score previously developed at our institution. A retrospective analysis of a random subset of 300 patients from a previously published randomized trial of critically ill adults was performed, with manual validation of SOFA scores from the date of initial intensive care unit admission. Spearman's rank correlation coefficient, weighted Cohen's kappa, and Bland-Altman plots were used to assess agreement between manual and electronic versions of SOFA scores and between manual and electronic versions of their individual components. There was high agreement between manual and electronic SOFA scores (Spearman's rank correlation coefficient = 0.90, 95% CI 0.87-0.93). Renal and respiratory components had lower agreement (weighted Cohen's kappa = 0.63, 95% CI 0.53-0.73 for renal; weighted Cohen's kappa = 0.77, 95% CI 0.70-0.84 for respiratory). The area under the receiver operating characteristic curve (AUC) for 30-day in-hospital mortality was 0.77 (95% CI 0.68-0.84) for manual SOFA scores and 0.75 (95% CI 0.66-0.83) for automated SOFA scores. Automatic calculation of SOFA scores from the electronic health record is feasible and correlates highly with manually calculated SOFA scores. Both have similar predictive value for 30-day in-hospital mortality.


Successful treatment of pustulosis palmaris et plantaris with granulocyte colony stimulating factor in a patient with hereditary neutropenia.
A 26-year-old female with hereditary neutropenia had Pustulosis Palmaris et Plantaris. Drug administration did not improve her symptoms. Following administration of granulocyte colony stimulating factor (G-CSF; filgrastim), her neutrophil count increased from 300 to 58,000/microliters, and her dermatosis improved. Pustulosis Palmaris et Plantaris has been described as a representative second lesion of focal infection. Neutropenia may be one cause of Pustulosis Palmaris et Plantaris due to refractory focal infection. In the present case, increase in neutrophils with G-CSF may have improved focal infection, resulting in improvement in refractory Pustulosis Palmaris et Plantaris.


Are APLS formulae for estimating weight appropriate for use in children admitted to PICU?
To determine if the revised APLS UK formulae for estimating weight are appropriate for use in the paediatric intensive care population in the United Kingdom. A retrospective observational study involving 10,081 children (5622 male, 4459 female) between the age of term corrected and 15 years, who were admitted to Paediatric Intensive Care Units in the United Kingdom over a five year period between 2006 and 2010. Mean weight was calculated using retrospective data supplied by the 'Paediatric Intensive Care Audit Network' and this was compared to the estimated weight generated using age appropriate APLS UK formulae. The formula 'Weight=(0.5×age in months)+4' significantly overestimates the mean weight of children under 1 year admitted to PICU by between 10% and 25.4%. While the formula 'Weight=(2×age in years)+8' provides an accurate estimate for 1-year-olds, it significantly underestimates the mean weight of 2-5 year olds by between 2.8% and 4.9%. The formula 'Weight=(3×age in years)+7' significantly overestimates the mean weight of 6-11 year olds by between 8.6% and 20.7%. Simple linear regression was used to produce novel formulae for the prediction of the mean weight specifically for the PICU population. The APLS UK formulae are not appropriate for estimating the weight of children admitted to PICU in the United Kingdom. Relying on mean weight alone will result in significant error as the standard deviation for all age groups are wide.


Syndrome of camptodactyly, ankyloses, facial anomalies, and pulmonary hypoplasia (Pena-Shokeir syndrome): obstetric and ultrasound aspects.
Two siblings with Pena-Shokeir syndrome are described. This syndrome consists of polyhydramnios, intrauterine growth retardation, short umbilical cord, perinatal death, facial abnormalities, limb abnormalities including arthrogryposis, and lethal pulmonary hypoplasia. The mode of inheritance is most likely autosomal recessive. Prenatal diagnosis was made in the second pregnancy with ultrasound performed at 26 weeks' gestation. The roles of fetal akinesia and fetal apnea in the production of the various manifestations of the syndrome are detailed, and the possibility of early prenatal diagnosis is considered.


Increased risk of developing affective disorder in patients with hypothyroidism: a register-based study.
Links between thyroid function and depression have been noted in many contexts. We assessed whether hospitalization with hypothyroidism was a risk factor for developing affective disorder. We conducted a prospective cohort study using historical data from Danish registers. The observational period was 1977-1999. Three study cohorts were identified: all patients with a first hospital admittance with the resulting index discharge diagnoses hypothyroidism, osteoarthritis, or nontoxic goiter. A later hospitalization with a resulting discharge diagnosis of affective disorder was used as event of interest, and rates of readmission were estimated and compared using competing risk models in survival analyses. We identified 165,307 patients discharged with an index diagnosis. In the observational period, 1041 events occurred. An index diagnosis of hypothyroidism was associated with an increased risk of hospitalization with affective disorder when compared to the control diseases. The risk of hospitalization with affective disorder was greatest in the first year after index hospitalization. Patients hospitalized with hypothyroidism have a greater risk of readmission with depression or bipolar disorder than control patients. This renders epidemiologic support to theories linking thyroid dysfunction with mood disturbances.


[Some measures of proliferative activity in normal and leukoplakic buccal mucosa (author's transl)].
In 29 patients, the DNA synthesis rate of clinically healthy buccal mucosa epithelium in comparison to benign buccal leukoplakias has been studied by in vitro-autoradiography. The proliferative activity has been determined by means of the 3H-labelling indices of basal cells (LBC) and suprabasal cells (LSBC) including the total of labelled epithelium cells and the quotient of LBC:LSBC as well. The progenitor compartment of both leukoplakic and normal buccal epithelium comprises the stratum basale and the adjacent 2-3 layers of suprabasal cells. Around 70% of the DNA synthesis of normal mucosa was found in the suprabasal nuclei of the progenitor compartment. In the leukoplakic mucosa, some displacement of the labelled nuclei in the progenitor compartment resulting in a statistically significant change of the quotient of LBC:LSBC in favour of LBC was determined. Moreover, the leukoplakic specimens showed a moderate decrease of the mean proliferative activity of the total of labelled epithelial cells which may be due to a diminished exfoliation of cornified cells from the hyper(ortho)keratotic surface. It is supposed that in chronic oral leukoplakias some variation of the steady state between renewal and desquamative loss of epithelial cells is operating as far as no precancerous condition is present.


Platelet-Rich Plasma: Review of Current Literature on its Use for Tendon and Ligament Pathology.
Platelet-rich plasma (PRP) contains numerous growth factors and cytokines that potentially offer an alternative treatment modality to assist in the healing of multiple musculoskeletal issues. The purpose of this review was to examine the latest literature on the use of PRP for various ligament and tendon pathologies. Recent literature has shown moderate- to high-quality evidence that PRP can have positive clinical effects in certain conditions such as lateral epicondylitis and rotator cuff tendinopathy. Prospective studies have shown that it can also be useful in the treatment of patella tendinopathy. In summary, we found PRP to have variable success in ligament and tendon pathology; however, it should be considered a viable option in chronic musculoskeletal disease that has failed other treatments. Patient selection, duration of symptoms, and combining with other modalities such as physical therapy should all be taken into consideration in treatment with PRP.


Totally Thoracoscopic Surgical Ablation or Catheter Ablation of Atrial Fibrillation: A Systematic Review and Preliminary Meta-Analysis.
This article provides results of a preliminary meta-analysis of the stand-alone atrial fibrillation (AF) surgical publications from 2009 to 2011. In 6 studies with postoperative freedom from AF as primary end point, the meta-analysis indicated an overall success rate of 84% (95% confidence interval [CI], 80.0-88.0). For the 7 studies with the primary end point of postoperative return to normal sinus rhythm, the success rate was 83% (95% CI, 79.0-87.0). These data are compared with 3 comprehensive meta-analyses of catheter ablation. Based on comparison, it is recommended that initial treatment of long-standing persistent atrial fibrillation be minimal access surgery.


Effect of Tricalcium Phosphate and Calcium Sucrose Phosphate on the Inhibition of Streptococcus mutans: An In Vivo Study.
To determine the efficacy of tricalcium phosphate (TCP) and calcium sucrose phosphate (CSP) on the inhibition of Streptococcus mutans (SM). Thirty healthy children between 13 and 18 years of age were divided into two groups of 15 each; Group I receiving TCP-containing cream and Group II receiving calcium sucrose phosphate-containing cream. On the first day of the study, 30 minutes after breakfast, baseline plaque samples were taken from the buccal surface of first mandibular permanent molar using a sterile wedge which was immediately transferred to sterile container containing 1 mL of saline, and were subjected to microbiological examination. On the following days, both the creams were applied to the respective groups. On the 16th day, plaque samples were collected from the same site, and colony forming units were recorded using agar plate as a culture medium. The mean of S. mutans count before application of TCP-containing paste was 16.27 cfu per mL and before calcium sucrose phosphate-containing paste was 15.33 cfu per mL. The mean after application of TCP-containing paste and calcium sucrose phosphate-containing paste was 3.53 and 9.87 cfu per mL, respectively. And, there was a statistically significant difference found within the groups. Both TCP and CSP have an inhibitory effect on S. mutans. This can be an effective preventive tool for children with high caries risk and even for special child. Both TCP and CSP deposit the mineral reservoir in plaque and saliva; it may help resist the future cariogenic challenges.


Lichen sclerosus et atrophicus.
A case of lichen sclerosus et atrophicus (LSA) involving the oral mucosa is described. The lesion began in the labial mucosa with extension to the gingiva and to the skin. The frequency of occurrence of oral lesions of LSA is discussed, and a histopathologic comparison with skin lesions is made.


Alterations in follicular IGFBP mRNA expression and follicular fluid IGFBP concentrations during the first follicle wave in beef heifers.
The objective was to determine the pattern of IGFBP-2, -3 and -4 gene expression and follicular fluid concentrations of IGFBP-2, -3, -4 and -5 during emergence, selection and dominance of the first follicle wave of the estrous cycle in cattle and during exogenous steroid treatment. Heifers (n = 35) were ovariectomized at 36 (n = 7), 66 (n = 8), 84 (n = 12) and 108 (n = 8) h after the onset of estrus. Heifers in the 84 h ovariectomy group were sub-divided to receive either no treatment (n = 6) or were treated with a progesterone-releasing intravaginal device (n = 6, PRID) and 0.75 mg estradiol benzoate i.m. at the approximate time of ovulation, 30 h post estrus until ovariectomy. Within heifers the four largest follicles recovered following ovariectomy were ranked on size (F1, F2, F3 and F4). At 36 h IGFBP gene expression and follicular fluid IGFBP concentrations were similar in all follicles (F1-F4). Mean diameter of the F1 follicle increased (P < 0.05) between 36 and 84 h with no difference between 84 and 108 h. The F1 follicle had the highest (P < 0.05) concentration of estradiol compared with the F2, F3 and F4 at 84 and 108 h. There was no granulosa cell IGFBP-2 mRNA in F1 follicles at 84 or 108 h. Intrafolliclar IGFBP-2 concentrations were lower (P < 0.05) in the F1 compared with F3 and F4 follicles at 108 h. There was no difference in theca cell IGFBP-4 mRNA expression at 108h, but amounts of follicular fluid IGFBP-4 were lower (P < 0.05) in F1 follicles compared with F3 and F4 follicles at 108 h. IGFBP-3 mRNA was localized in the theca layer of all follicles examined with no difference in expression or follicular fluid concentrations during emergence, selection and dominance of the first follicle wave. IGFBP-5 concentrations were higher (P < 0.05) in follicular fluid of F3 follicles at 108 h compared with the F3 at 36 h. In conclusion follicular dominance was associated with low or decreased follicular fluid concentrations of IGFBP-4 and -5, increased estradiol and differential regulation of IGFBP production.


Graph-Based Reconstruction and Analysis of Disease Transmission Networks Using Viral Genomic Data.
Understanding the patterns of viral disease transmissions helps establish public health policies and aids in controlling and ending a disease outbreak. Classical methods for studying disease transmission dynamics that rely on epidemiological data, such as times of sample collection and duration of exposure intervals, struggle to provide desired insight due to limited informativeness of such data. A more precise characterization of disease transmissions may be acquired from sequencing data that reveal genetic distance between viral genomes in patient samples. Indeed, genetic distance between viral strains present in hosts contains valuable information about transmission history, thus motivating the design of methods that rely on genomic data to reconstruct a directed disease transmission network, detect transmission clusters, and identify significant network nodes (e.g., super-spreaders). In this article, we present a novel end-to-end framework for the analysis of viral transmissions utilizing viral genomic (sequencing) data. The proposed framework groups infected hosts into transmission clusters based on the reconstructed viral strains infecting them; the genetic distance between a pair of hosts is calculated using Earth Mover's Distance, and further used to infer transmission direction between the hosts. To quantify the significance of a host in the transmission network, the importance score is calculated by a graph convolutional autoencoder. The viral transmission network is represented by a directed minimum spanning tree utilizing the Edmond's algorithm modified to incorporate constraints on the importance scores of the hosts. The proposed framework outperforms state-of-the-art techniques for the analysis of viral transmission dynamics in several experiments on semiexperimental as well as experimental data.


Expression of c-fos and other genes encoding transcription factors in long-term potentiation.
The genes encoding transcription factors are known to be induced in many different biological phenomena of transition from one long-lasting state of cell functioning to another. It is widely believed that transcription factors control this transition by regulating the expression of other genes. Recently, several reports on gene expression after the induction of long-term potentiation (LTP) have been published. In particular, the c-fos nuclear protooncogene, encoding the Fos transcription factor, has been extensively investigated. However, the results of those studies were seemingly contradictory. The present commentary reviews available data in an attempt to resolve the apparent contradictions, showing that long-lasting LTP (i.e., lasting longer than a few hours) may involve c-fos expression, while shorter LTP may not.


A deep connectome learning network using graph convolution for connectome-disease association study.
Multivariate analysis approaches provide insights into the identification of phenotype associations in brain connectome data. In recent years, deep learning methods including convolutional neural network (CNN) and graph neural network (GNN), have shifted the development of connectome-wide association studies (CWAS) and made breakthroughs for connectome representation learning by leveraging deep embedded features. However, most existing studies remain limited by potentially ignoring the exploration of region-specific features, which play a key role in distinguishing brain disorders with high intra-class variations, such as autism spectrum disorder (ASD), and attention deficit hyperactivity disorder (ADHD). Here, we propose a multivariate distance-based connectome network (MDCN) that addresses the local specificity problem by efficient parcellation-wise learning, as well as associating population and parcellation dependencies to map individual differences. The approach incorporating an explainable method, parcellation-wise gradient and class activation map (p-GradCAM), is feasible for identifying individual patterns of interest and pinpointing connectome associations with diseases. We demonstrate the utility of our method on two largely aggregated multicenter public datasets by distinguishing ASD and ADHD from healthy controls and assessing their associations with underlying diseases. Extensive experiments have demonstrated the superiority of MDCN in classification and interpretation, where MDCN outperformed competitive state-of-the-art methods and achieved a high proportion of overlap with previous findings. As a CWAS-guided deep learning method, our proposed MDCN framework may narrow the bridge between deep learning and CWAS approaches, and provide new insights for connectome-wide association studies.


Identification of a Transmembrane Protein Involved in Shear Stress Signaling and Hepatocarcinogenesis After a Sustained Virological Response to Hepatitis C Virus.
The risk of hepatocellular carcinoma (HCC) remains after achieving a sustained virological response (SVR) in patients with chronic hepatitis C (CHC). Epigenetic abnormalities might be key regulators in the development of HCC. This study aimed to identify the genes involved in hepatocarcinogenesis after an SVR. DNA methylation in liver tissue was compared between 21 CHC patients without HCC and 28 CHC patients with HCC, all of whom had achieved an SVR. Additional comparisons with 23 CHC patients before treatment and 10 normal livers were performed. The characteristics of a newly identified gene were explored in vitro and in vivo. We found that the transmembrane protein no. 164 (TMEM164) gene was demethylated by hepatitis C virus infection and HCC development after achieving an SVR. TMEM164 was expressed mainly in endothelial cells, alpha smooth muscle actin-positive cells, and some capillarized liver sinusoidal endothelial cells. TMEM164 expression was significantly correlated with liver fibrosis and relapse-free survival in HCC patients. TMEM164 was induced by shear stress, interacted with GRP78/BiP, accelerated ATF6 (activating transcription factor 6)-mediated endoplasmic reticulum (ER) stress signaling, and activated interleukin-6/STAT3 (signal transducer and activator of transcription 3) signaling in the TMNK1 liver endothelial cell line. Therefore, we termed TMEM164 "shear stress-induced transmembrane protein associated with ER stress signaling" (SHERMER). SHERMER knockout mice were protected against CCL4-induced liver fibrosis. SHERMER overexpression in TMNK1 cells accelerated HCC growth in a xenograft model. We identified a new transmembrane protein, SHERMER, in CHC patients with HCC after achieving an SVR. SHERMER was induced by shear stress and accelerated ATF6-mediated ER stress signaling in endothelial cells. Thus, SHERMER is a novel endothelial marker associated with liver fibrosis, hepatocarcinogenesis, and progression of HCC.


Condensation pattern (CP) analysis of plant chromosomes by an improved chromosome image analysing system, CHIAS III.
The third-generation chromosome image analysing system (CHIAS III) was developed by using an ordinary personal computer with public domain imaging software, the NIH Image. An uneven condensation pattern (CP) appearing on small sugarcane chromosomes at the mitotic prometaphase stage was analysed and a quantitative chromosome map or idiogram was developed by using CHIAS III. The analysis consists of three major steps as follows: (1) manipulation of original chromosome images and measurement of CP or numerical data of the CP; (2) calculation of an averaged CP for homologous chromosomes; and (3) development of an idiogram based on the averaged CP. The CP has been analysed quantitatively by using the first- and the second-generation chromosome image analysing systems with hard- and software specific to the systems. The data obtained using CHIAS III are consistent with those obtained with the chromosome image analysing systems previously developed.


The discovery of potent and selective pyridopyrimidin-7-one based inhibitors of B-RafV600E kinase.
Herein we describe the discovery of a novel series of ATP competitive B-Raf inhibitors via structure based drug design (SBDD). These pyridopyrimidin-7-one based inhibitors exhibit both excellent cellular potency and striking B-Raf selectivity. Optimization led to the identification of compound 17, a potent, selective and orally available agent with excellent pharmacokinetic properties and robust tumor growth inhibition in xenograft studies.


Status epilepticus and tiagabine therapy revisited.
To determine whether antiepileptic treatment with tiagabine (TGB) is associated with an increased frequency of nonconvulsive status epilepticus (NCSE) in patients with refractory epilepsy. We reviewed retrospectively the medical and EEG records of all inpatients with refractory localization-related epilepsy at the National Society for Epilepsy treated with TGB between January 1997 and December 2000. Clinical and electroencephalographic (EEG) data before, during, and after TGB therapy were evaluated in those patients who experienced a deterioration in seizure control suggestive of NCSE. Frequency of NCSE was determined in a comparable, non-TGB-treated patient population. Seven (7.8%) of 90 TGB-treated patients were identified who experienced episodes of electroclinically confirmed NCSE. Serial EEGs showed deterioration during TGB treatment, with resolution of abnormality on discontinuation of TGB in all seven patients. During the same observation period, 32 (2.7%) of 1,165 non-TGB-treated patients developed electroclinically defined NCSE. Treatment with TGB is associated with an increased frequency of NCSE in patients with refractory localization-related epilepsy.


Electrospray ionization tandem mass spectrometry of sodiated adducts of cholesteryl esters.
Cholesteryl esters (CE) are important lipid storage molecules. The present study demonstrates that sodiated adducts of CE molecular species form positive ions that can be detected in both survey scan mode as well as by exploiting class-specific fragmentation in MS/MS scan modes. A common neutral loss for CE is the loss of cholestane (NL 368.5), which can be used to specifically quantify tissue CE molecular species. Using this MS/MS technique, CE molecular species were quantified in mouse monocyte-derived macrophages (J774 cells) incubated with either linoleic (18:2) or arachidonic acid (20:4). These studies revealed that arachidonic acid was not only incorporated into the CE pool, but also was elongated resulting in the accumulation of 22:4 and 24:4 CE molecular species in macrophages. Additionally, this technique was used to quantify CE molecular species present in crude lipid extracts from plasma of female mice fed a Western diet, which led to an enrichment in CE molecular species containing monounsaturated fatty acids compared to female mice fed a normal chow diet. Last, NL 368.5 spectra revealed the oxidation of the aliphatic fatty acid residues of CE molecular species containing polyunsaturated fatty acids. Taken together, these studies demonstrate the utility of using sodiated adducts of CE in conjunction with direct infusion electrospray ionization tandem mass spectrometry to rapidly quantify CE molecular species in biological samples.


Characteristics of anion transport in cat and dog red blood cells.
Self-exchange of chloride and sulfate in dog and cat red cells has been measured under equilibrium conditions. The rats of efflux for these anions are approximately twofold higher in dog compared to cat red blood cells. Although the rates differ, the anion exchange systems of these two red cell types exhibit many common properties. The dependence of 35SO4 efflux on the intracellular SO4 concentration, the pH dependence and the inhibition of 35SO4 efflux by Cl and SITS are almost identical in dog and cat red cells. Nystatin treatment was used to study the dependence of 36Cl efflux on internal Cl. Chloride efflux exhibits saturation in both cell types with dog red cells possessing a higher Vmax and K1/2 than cat red cells. The number of anion transport sites was estimated by extrapolation to the number of molecules of dihydro DIDS (H2DIDS, where DIDS is 4,4'-diisothiocyano-2,2' stilbene-disulfonic acid) which were bound at 100% inhibition of transport. The results indicate that either the turnover numbers for anion transport differ in dog, cat, and human red cells or that there is heterogeneity in the function of the membrane components which bind H2DIDS.


Interindividual variation in the capacity-limited renal glucuronidation of probenecid by humans.
A dose of 1,000 mg probenecid was administered orally to 14 human volunteers in order to quantify the maximal rate of formation and excretion of probenecid acyl glucuronide in the urine. Probenecid showed dose-dependent pharmacokinetics. Plasma protein binding of probenecid was high, being somewhat higher in males (90.7 +/- 1.4%) than in females (87.9 +/- 1.4%; p = 0.0019). It was shown that probenecid is metabolized by cytochrome P-450 into at least two phase I metabolites. Each of the metabolites accounted for less than 12% of the dose administered; the main metabolite probenecid acyl glucuronide, representing 42.9 +/- 13.2% of the dose, was only present in urine and not in plasma. The renal excretion rate-time profile of probenecid acyl glucuronide showed a plateau value in the presence of an acidic urine pH. This plateau value was maintained for about 10 h at the dose of 1,000 mg. The height of the plateau value depended on the individual and varied between 250 and 800 micrograms/min (15-50 mg/h). It was inferred that probenecid acyl glucuronide is formed in the kidney during blood-to-lumen passage through the tubular cells. We conclude that the plateau value in the renal excretion rate of probenecid glucuronide reflects its Vmax of formation.


Proteinase-activated receptor-1-triggered activation of tumor progression locus-2 promotes actin cytoskeleton reorganization and cell migration.
Tumor progression locus 2 (Tpl2), a mitogen-activated protein kinase kinase kinase (MAP3K) that is activated by provirus insertion in retrovirus-induced rodent lymphomas and mammary adenocarcinomas, is known to transduce Toll-like receptor, interleukin 1, tumor necrosis factor alpha, and CD40 signals and to play an important role in inflammation. Here we show that Tpl2 is also required for the transduction of cell migration and gene expression signals originating in the G-protein-coupled receptor proteinase-activated receptor 1 (PAR1). PAR1 signals transduced by Tpl2 activate Rac1 and focal adhesion kinase, and they are required for reorganization of the actin cytoskeleton and cell migration. PAR1 expressed in fibroblasts can be triggered by proteinases produced by tumor cells, and PAR1 expressed in tumor cells can be triggered by proteinases produced by fibroblasts. These data suggest that signals that regulate cell migration and gene expression flow between stromal and tumor cells in both directions and that Tpl2 plays a pivotal role in this process.


Targeted gold-coated iron oxide nanoparticles for CD163 detection in atherosclerosis by MRI.
CD163 is a membrane receptor expressed by macrophage lineage. Studies performed in atherosclerosis have shown that CD163 expression is increased at inflammatory sites, pointing at the presence of intraplaque hemorrhagic sites or asymptomatic plaques. Hence, imaging of CD163 expressing macrophages is an interesting strategy in order to detect atherosclerotic plaques. We have prepared a targeted probe based on gold-coated iron oxide nanoparticles vectorized with an anti-CD163 antibody for the specific detection of CD163 by MRI. Firstly, the specificity of the targeted probe was validated in vitro by incubation of the probe with CD163(+) or (-) macrophages. The probe was able to selectively detect CD163(+) macrophages both in human and murine cells. Subsequently, the targeted probe was injected in 16 weeks old apoE deficient mice developing atherosclerotic lesions and the pararenal abdominal aorta was imaged by MRI. The accumulation of probe in the site of interest increased over time and the signal intensity decreased significantly 48 hours after the injection. Hence, we have developed a highly sensitive targeted probe capable of detecting CD163-expressing macrophages that could provide useful information about the state of the atheromatous lesions.


Ex vivo infection of murine precision-cut lung tissue slices with Mycobacterium abscessus: a model to study antimycobacterial agents.
Multidrug-resistant infections due to Mycobacterium abscessus often require complex and prolonged regimens for treatment. Here, we report the evaluation of a new ex vivo antimicrobial susceptibility testing model using organotypic cultures of murine precision-cut lung slices, an experimental model in which metabolic activity, and all the usual cell types of the organ are found while the tissue architecture and the interactions between the different cells are maintained. Precision cut lung slices (PCLS) were prepared from the lungs of wild type BALB/c mice using the Krumdieck® tissue slicer. Lung tissue slices were ex vivo infected with the virulent M. abscessus strain L948. Then, we tested the antimicrobial activity of two drugs: imipenem (4, 16 and 64 μg/mL) and tigecycline (0.25, 1 and 4 μg/mL), at 12, 24 and 48 h. Afterwards, CFUs were determined plating on blood agar to measure the surviving intracellular bacteria. The viability of PCLS was assessed by Alamar Blue assay and corroborated using histopathological analysis. PCLS were successfully infected with a virulent strain of M. abscessus as demonstrated by CFUs and detailed histopathological analysis. The time-course infection, including tissue damage, parallels in vivo findings reported in genetically modified murine models for M. abscessus infection. Tigecycline showed a bactericidal effect at 48 h that achieved a reduction of > 4log10 CFU/mL against the intracellular mycobacteria, while imipenem showed a bacteriostatic effect. The use of this new organotypic ex vivo model provides the opportunity to test new drugs against M. abscessus, decreasing the use of costly and tedious animal models.


Stereoselective synthesis of chiral, non-racemic 1,2,3-tri- and 1,3-disubstituted ferrocene derivatives.
Chiral, non-racemic 1,2,3-trisubstituted ferrocene derivatives are accessible from monosubstituted ferrocenes through two sequential ortho-deprotonation reactions; removal of the central substituent gives 1,3-disubstituted ferrocenes.


Genomic regulation of intestinal amino acid transporters by aldosterone.
Overexpression of renal LAT2, a Na+ -independent L-amino acid transporter, in spontaneous hypertensive rats (SHR) is organ specific and precedes the onset of hypertension (Pinho et al., Hypertension, 42:613-618, 2003). However, the expression of LAT2 correlates negatively with plasma aldosterone levels after high sodium intake (Pinho et al., Am J Physiol Ren Physiol 292:F1452-F1463, 2007). The present study evaluated the expression of Na+ -independent LAT1, LAT2, and 4F2hc and Na+ -dependent ASCT2 amino acid transporters in the intestine of normotensive Wistar rats chronically treated with aldosterone. In conditions of high salt intake, to keep endogenous aldosterone to a minimum, rats were implanted with aldosterone or spironolactone tablets. In aldosterone-treated and aldosterone + spironolactone-treated rats, aldosterone plasma levels were increased by fourfold. At the protein level, aldosterone treatment significantly increased LAT1 (62%), LAT2 (49%), 4F2hc (48%), and ASCT2 (65%) expression. The effect of aldosterone upon LAT1, LAT2, 4F2hc, and ASCT2 protein abundance was completely reversed by spironolactone. Aldosterone significantly increased intestinal LAT2 and 4F2hc mRNA levels (27% and 35% increase, respectively), with no changes in LAT1 and ASCT2 transcript levels. In conclusion, increases in intestinal Na+ -independent LAT1 and LAT2 and Na+ -dependent ASCT2 transcript and protein abundance during chronic treatment with aldosterone occur through a spironolactone-sensitive genomic mechanism.


Influence of intravesical administration of tumor necrosis factor alpha and systemic gamma-interferon on murine bladder cancer: lack of dose response.
The effect of intravesical administration of high dose recombinant tumor necrosis factor-alpha (rTNF) and in combination with systemic recombinant gamma-interferon (rIFN) on murine bladder cancer was studied. RTNF was given at 12.5 micrograms/mouse on days 7, 11 and 15 after tumor instillation or at 2.5 micrograms/mouse on days 7, 9, 11, 13 and 15. Some groups were also injected i.v., 24-h prior to each rTNF treatment with rIFN at a dose of 1.3 micrograms/mouse. RTNF treatment suppressed tumor growth up to 48% of control, although the difference was not statistically significant. Combined administration of rIFN did not provide additional benefit.


A new genus and species of Nannopodidae (Crustacea, Copepoda, Harpacticoida) from the Yellow Sea of South Korea.
A new monospecific genus of the family Nannopodidae Brady, 1880 is proposed, based on specimens of both sexes of Concilicoxa hispida gen. et sp. nov. collected from subtidal sandy sediments in the Yellow Sea of South Korea. The presence of a coxal outer projection on the first to fourth legs and reduction of both rami of the second to fourth legs in this new genus show a clear relationship with a clade, which is characterised by the modified thoracopods for burrowing ability, comprising Huntemannia Poppe, 1884, Rosacletodes Wells, 1985, Laophontisochra George, 2002, Acuticoxa Huys & Kihara, 2010 and Talpacoxa Corgosinho, 2012 in Nannopodidae. Within this clade, C. hispida gen. et sp. nov. is most closely related to L. maryamae George, 2002 in having the prehensile endopod in the first leg, broad intercoxal sclerite on the second to fourth legs and the female fifth leg being composed of separate exopod and baseoendopod, but is distinguished by the absence of mandibular exopod, two-segmented mandibular endopod, presence of four setae on the distal exopodal segment of the first leg, and fusion of the intercoxal sclerite to the coxae in the third and fourth legs. These four features are considered as autapomorphies of the new genus. The possible relationship amongst members of the nannopodid clade is further discussed. Additionally, some comments on the taxonomic position of L. terueae Björnberg, 2014 are given, resulting in the transfer of the species to Acuticoxa as A. terueae comb. nov.


P4HA2-induced prolyl hydroxylation of YAP1 restricts vascular smooth muscle cell proliferation and neointima formation.
Vascular smooth muscle cell (VSMC) proliferation and neointima formation play significant roles in atherosclerosis development and restenosis following percutaneous coronary intervention. Our team previously discovered that TEA domain transcription factor 1 (TEAD1) promotes vascular smooth muscle differentiation, which is necessary for vascular development. Conversely, aberrant YAP1 activation upregulates the platelet-derived growth factor receptor beta to encourage VSMC proliferation and neointima formation. In this study, we aimed to investigate the molecular mechanisms of YAP1/TEAD signaling during neointima formation. Our research focused on the prolyl 4-hydroxylase alpha 2 (P4HA2) and its downstream target, Yes-associated protein 1 (YAP1), in regulating VSMC differentiation and neointima formation. Our results indicated that P4HA2 reduction leads to VSMC dedifferentiation and promotes neointima formation after injury. Furthermore, we found that P4HA2-induced prolyl hydroxylation of YAP1 restricts its transcriptional activity, which is essential to maintaining VSMC differentiation. These findings suggest that targeting P4HA2-mediated prolyl hydroxylation of YAP1 may be a promising therapeutic approach to prevent injury-induced neointima formation in cardiovascular disease.