Donor-derived cell-free DNA (dd-cfDNA) for detection of allograft rejection in pediatric kidney transplants.
In pediatric transplantation, acute rejection is a major contributor of graft failure. Current approaches include kidney biopsy in response to graft dysfunction and/or the emergence of donor-specific HLA antibodies (DSA). However, biopsy is associated with potential complications. Thus, there is a need for non-invasive diagnostics. Detection of donor-derived cell-free DNA (dd-cfDNA, AlloSure) > 1% is associated with rejection in adult kidney transplants. Here, we evaluate the utility of dd-cfDNA for identifying allograft rejection in pediatric patients. Between 10/2017 and 10/2019, 67 patients, who underwent initial testing with dd-cfDNA as part of routine monitoring or in response to clinical suspicion for rejection, were included. Biopsies were performed when dd-cfDNA > 1.0% or where clinical suspicion was high. Demographics, dd-cfDNA, antibody status, and biopsies were collected prospectively. Data were analyzed to determine predictive value of dd-cfDNA for identifying grafts at risk for rejection. 19 of 67 patients had dd-cfDNA testing as part of routine monitoring with a median dd-cfDNA score of 0.37 (IQR: 0.19-1.10). 48 of 67 patients who had clinical suspicion of rejection had median dd-cfDNA score of 0.47 (0.24-2.15). DSA-positive recipients had higher dd-cfDNA scores than those who were negative or had AT1R positivity alone (P = .003). There was no association between dd-cfDNA score and strength of DSA positivity. 7 of 48 recipients had a biopsy with a dd-cfDNA score <1%; two showed evidence of rejection. Neither DSA nor AT1R positivity was statistically associated with biopsy-proven rejection. However, dd-cfDNA >1% was diagnostic of rejection with sensitivity of 86% and specificity of 100% (AUC: 0.996, 0.98-1.00; P = .002). dd-cfDNA represents a non-invasive method for early detection of rejection in pediatric renal transplants. Our study shows dd-cfDNA to be highly predictive of histological rejection and superior to other indicators such as graft dysfunction or antibody positivity alone. Further studies are necessary to refine these initial observations.


Somatostatin receptor positron emission tomography/computed tomography imaging in Merkel cell carcinoma.
Merkel cell carcinoma (MCC) is an uncommon aggressive primary cutaneous carcinoma with neuroendocrine differentiation. However, literature data about the use of somatostatin receptor positron emission tomography/computed tomography (PET/CT) imaging in MCC are limited and its role is not clearly stated. To investigate the role of PET/CT using somatostatin analogues radiolabelled with gallium-68 in patients with MCC. All patients affected by MCC who performed a somatostatin receptor PET/CT imaging from October 2007 to May 2014 were retrospectively analysed. The diagnostic performances of PET/CT were evaluated on a patient-based analysis and compared to final diagnosis (histology = 3 or clinical/radiological follow-up = 20). We evaluated 23 consecutive MCC patients [18 men; median age 71 years (range 47-87)]. Primary tumour was located in ear (1/23), cheek (3/23), arm (2/23), hand (1/23), back (1/23), anal canal (1/23), gluteus (4/23), thigh (3/23) and popliteal fossa (1/23). In 6/23 patients, the site of primary tumour was unknown. PET/CT was performed to detect primary tumour site (4/23) or to stage (8/23) or re-stage (11/23) patients. PET/CT resulted positive in 14/23 patients and according to the final diagnosis was defined true positive, true negative, false positive (FP) and false negative in 11/23, 8/23, 3/23 and 1/23 cases respectively. FP PET/CT results were due to unspecific liver uptake, post-surgical inflammation and pancreatic neuroendocrine tumour. PET/CT was unable to detect primary tumour site in all patients with unknown primary MCC. Sensitivity, specificity and diagnostic accuracy of PET/CT were 92%, 73% and 83% respectively. In our experience, somatostatin receptor PET/CT imaging resulted useful in patients with MCC and presented high diagnostic performances with a significant impact in disease management although in patients with unknown primary MCC, it was unable to identify the primary tumour site.


Assessing psychopathology in bariatric surgery candidates: discriminant validity of the SCL-90-R and SCL-K-9 in a large sample of patients.
Pre-surgical psychosocial evaluation of bariatric surgery (BS) patients should identify psychiatric issues that could worsen after surgery and those requiring additional ongoing intervention. In this view, the use of reliable, appropriate and concise evaluating instruments is of critical importance. The aim of the present study was to investigate the clinical utility of both the Symptom Checklist 90-Revised (SCL-90-R) and its brief unidimensional version, the so-called Symptom-Checklist-K-9 (SCL-K-9) in detecting the presence of psychiatric disorders among bariatric surgery (BS) candidates. Seven-hundred-and-ninety-eight BS candidates (563 women and 235 men; mean age: 44.15 ± 11.45) were enrolled in the present study. All participants underwent a full psychiatric interview and were administered the SCL-90-R. Three-hundred-and-sixty-two patients (45.4%) met the criteria for a diagnosis of at least one psychiatric disorder and ninety-nine patients (12.4%) had psychiatric comorbidities. In the current sample, 219 patients (27.4%) met the criteria for binge eating disorders (BED), 158 (19.8%) met the criteria for major depressive disorder (MDD), and 67 (8.4%) met both criteria. A receiver operating characteristic (ROC) curves procedure showed that both the SCL-90-R and the SCL-K-9 satisfactorily categorize patients with any psychiatric disorder, both BED and MDD (area under the ROC curve ≥ 0.70, p < 0.001). Our results suggest that the SCL-90-R and the SCL-K-9 may represent first-level screening tests identifying at-risk patients, eligible for a more expensive or time-consuming clinical assessment. Level V, cross-sectional, descriptive study.


Pharmacokinetics of benapenem for injection in subjects with mild to moderate renal impairment.
This study evaluated the pharmacokinetic (PK) characteristics of benapenem in subjects with mild to moderate renal impairment to provide a reference for benapenem dosing regimens in this patient population. Eighteen subjects were enrolled in this study. Each subject received a single dose of benapenem intravenously (1.0 g in 100 ml of 0.9% saline) followed by blood and urine collection to measure the concentrations of benapenem and its major metabolite. PK analysis was performed to evaluate the effect of varying degrees of renal impairment on the PK characteristics of benapenem. The safety of benapenem was also evaluated. In subjects with normal renal function, mild renal impairment, and moderate renal impairment, the maximum plasma benapenem concentrations were 163 ± 6.58 mg/L, 138 ± 17.4 mg/L, and 134 ± 0.11 mg/L, respectively (15.3% and 17.8% lower in subjects with mild and moderate renal impairment, respectively, than in subjects with normal renal function). The areas under the plasma concentration-time curve (AUC0-inf) were 1153.67 ± 143.2 mg·h/L, 1129.17 ± 241.41 mg·h/L, and 1316.46 ± 229.83 mg·h/L, respectively (P > 0.05); the cumulative urinary excretion rates at 72 h after dosing were 52.61 ± 8.58%, 39.42 ± 8.35%, and 29.84 ± 9.15%, respectively; and the metabolic ratio (AUC0-inf_KBP-3331/AUC0-inf_benapenem) were 3.96 ± 0.35%, 5.56 ± 0.82%, and 8.24 ± 0.85%, respectively. No drug-related adverse events (AEs), serious AEs, or AEs leading to withdrawal occurred in this study. No adjustment to benapenem dosing is needed in patients with mild to moderate renal impairment. Drug clinical trial registration and information publicity platform: http://www.chinadrugtrials.org.cn/index.html . CTR20190760.


Predicting the adaptive evolution of Thermoanaerobacterium saccharolyticum.
A fully evolved metabolic network can be described as a weighted sum of elementary modes where the usage probabilities of modes are distributed according to the Boltzmann distribution law (Srienc and Unrean, 2010). An organism presumably achieves the fully evolved state through adaptive changes in the kinetics of rate-controlling enzymes. Metabolic control analysis identifies reactions catalyzed by such enzymes. Comparison of the experimentally determined metabolic flux distributions of Thermoanaerobacterium saccharolyticum AS411 with the predicted flux distribution of a fully evolved metabolic network identified phosphoglucose isomerase (PGI) as the enzyme with the greatest flux control, the rate-controlling enzyme. The analysis predicts that an increased activity of PGI would enable the metabolic network to approach the fully evolved state and result in a faster specific growth rate. The prediction was confirmed by experimental results that showed an increased specific activity of PGI in a culture of strain AS411 that adaptively evolved over 280 generations. Sequencing of the gene confirmed the occurrence of a group of mutations clustered in the subunit binding domain of the dimeric enzyme. The results indicate that the evolutionary path is predictable as the strain AS411 adapted toward the fully evolved state by increasing the PGI activity. This experimental finding confirms that enzymes with predicted highest metabolic flux control are the targets of adaptive metabolic pathway evolution.


Synthesis, characterization and SERS activity of Au-Ag nanorods.
The formation mechanism and morphology of Au-Ag bimetallic colloidal nanoparticles depend on the composition. Ag coated Au colloidal nanoparticles have been prepared by deposition of Ag through chemical reduction on performed Au colloid. The composition of the Au100-x-Agx particles was varied from x=0 to 50. The obtained colloids were characterized by UV-vis spectroscopy and transmission electron microscopy (TEM). The Au80-Ag20 colloid consists of alloy nanorods with dimension of 25nm x 100nm. The activity of these nanorods in surface enhanced Raman spectroscopy (SERS) was checked by using sodium salicylate as an adsorbate probe. Intense SERS bands are observed indicating its usefulness as a SERS substrate in near infrared (NIR) laser excitation.


Rat Mrp2 gene expression is regulated by an interleukin-1β-stimulated biphasic response with enhanced transcription and subcellular shuttling of YB-1.
Expression of hepatobiliary transporters is decreased during endotoxemia. Reduction of Mrp2 is mediated by IL-1β-dependent signals but underlying mechanisms are still unclear. YB-1 is a predominantly cytoplasmic protein that translocates to the nucleus in response to various stimuli. Previously we have shown that YB-1 down-regulates Mrp2 expression in vitro. Therefore we investigated the potential role of YB-1 as regulator of hepatic acute phase genes. Liver sections from LPS-injected rats (20 h) were stained with YB-1-specific antibodies. Real-time RT-PCR quantification was performed for Mrp2, MMP-2 and YB-1. YB-1 protein was quantified from IL-1β- or TNFα-stimulated rat hepatoma cells (FaO) and the localization of a YFP-YB-1-CFP fusion protein was visualized by confocal microscopy in HepG2 human hepatocellular carcinoma cells. ChIP-assays and EMSA were performed to analyze YB-1 binding to DNA promoter elements. In endotoxemic livers Mrp2 mRNA was down-regulated by 80%, while YB-1 mRNA expression increased 2.5-fold. Immunohistochemical staining showed a marked up-regulation and predominant nuclear localization of YB-1 protein in LPS challenged rats. In FAO cells IL-1β incubation increased cytoplasmic YB-1 protein content up to 16h. IL-1β stimulation resulted in a 6-fold up-regulation of endogenous YB-1 in the nuclear compartment, which occurred within 90min. In accord with these findings nuclear fluorescence was detected with a YFP-YB-1-CFP fusion protein introduced in HepG2 cells. In addition to DNA binding studies with endotoxemic rat liver tissue, ChIP assays revealed an IL-1β-dependent increase of YB-1 binding to the Mrp2-promoter in FAO cells. YB-1 is activated during the hepatic acute phase response. IL-1β promotes a rapid nuclear YB-1 protein shuttling in hepatoma cells within 90 min and a transcriptional induction thereafter. This biphasic response may explain the IL-1β-mediated suppression of Mrp2 expression in endotoxemic rats.


Contamination of runoff water at Gdańsk Airport (Poland) by polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs).
Airport runoff can contain high concentrations of various pollutants, in particular polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), the environmental levels of which have to be monitored. Airport runoff water samples, collected at the Gdańsk-Rębiechowo Airport from 2008 to 2009, were analysed for PAHs and PCBs by gas chromatography. The aromatic fractions were separated by liquid-liquid extraction and analysed by GC/MS. Total PAH concentrations were 295-6,758 ng/L in 2008 and 180-1,924 ng/L in 2009, while total PCB levels in 2008 ranged from 0.14 to 0.44 μg/L and in 2009 from 0.06 to 0.23 μg/L. The PAH and PCB compositions in airport runoff waters were examined over a range of spatial and temporal scales to determine distributions, trends and possible sources. This pollution is mainly pyrolytic and related to anthropogenic activity. There were significant differences between the samples collected in the two seasons. An understanding of the magnitude of contamination due to airport runoff water is important for the effective management of airport infrastructure.


Lattice Dynamics Study of Phonon Instability and Thermal Properties of Type-I Clathrate K₈Si46 under High Pressure.
For a further understanding of the phase transitions mechanism in type-I silicon clathrates K₈Si46, ab initio self-consistent electronic calculations combined with linear-response method have been performed to investigate the vibrational properties of alkali metal K atoms encapsulated type-I silicon-clathrate under pressure within the framework of density functional perturbation theory. Our lattice dynamics simulation results showed that the pressure induced phase transition of K₈Si46 was believed to be driven by the phonon instability of the calthrate lattice. Analysis of the evolution of the partial phonon density of state with pressure, a legible dynamic picture for both guest K atoms and host lattice, was given. In addition, based on phonon calculations and combined with quasi-harmonic approximation, the specific heat of K₈Si46 was derived, which agreed very well with experimental results. Also, other important thermal properties including the thermal expansion coefficients and Grüneisen parameters of K₈Si46 under different temperature and pressure were also predicted.


Delayed type hypersensitivity (DTH) in anti-IgM-treated B cell-depleted mice: analysis of induction and effector phase.
The induction and the effector phase of murine delayed type hypersensitivity (DTH) were evaluated in mice treated from birth with anti-IgM antibodies; these mice had no mature B cells and could not produce an antibody response. To study the effector phase, long-term cultured cloned helper T cells were injected subcutaneously together with the specific antigen into the hind footpad of normal and B cell-deficient mice. Antigen-specific DTH responses assessed by the local swelling reaction 24 h after transfer measured against a particulate antigen (sheep red blood cells, SRBC) as well as a soluble antigen (ovalbumin, OVA) were unaffected by the absence of B cells. To study the induction phase of DTH, 3-day immune in vivo primed lymphocytes from normal or B cell-depleted mice were adoptively transferred by subcutaneous injection into the hind footpad of naive syngeneic recipients. B cell depletion did not affect the induction of cells capable of responding to SRBC; in contrast, the response to soluble antigen (OVA) was significantly reduced, suggesting that B cells or their products participated in the induction of a DTH response to a soluble antigen.


Bone water.
A short review is given of the water in bone. Various analyses of bone water content are discussed, and its possible location is considered in relation to the behaviour of water in isolated components of bone. Some of the difficulties encountered in examining such microscopic phenomena as water structure in a heterogeneous system such as bone are also discussed.


Titrating matching-to-sample performance in pigeons: effects of diazepam, morphine, and cholinergic agents.
Five adult, male White Carneau pigeons were trained to respond under a titrating matching-to-sample schedule of reinforcement. Under this titration schedule, each trial began with the presentation of a sample stimulus (red or green light) on the center key of a three-key pigeon chamber. Completion of 15 responses on the center key resulted in the termination of the stimulus presentation and the initiation of a delay period. The length of the delay changed as a function of the pigeon's performance. During the first five trials of each session, the delay was fixed at 3 s in length. On the sixth and all subsequent trials, the length of the delay was either increased, did not change, or decreased such that accuracy was maintained at approximately 80%. Following the delay, two of the three pigeon keys were transilluminated with different colored lights (red or green). A single response upon the key transilluminated with the same stimulus color as the sample stimulus resulted in the presentation of food. A response on the key transilluminated with the stimulus color that did not match the sample stimulus resulted in a time-out period. Using this procedure, the effects of two drugs of abuse, diazepam (0.03-3 mg/kg) and morphine (0.03-10 mg/kg), a muscarinic antagonist, scopolamine (0.003-0.3 mg/kg), the quaternary derivative of scopolamine, methylscopolamine (0.003-0.3 mg/kg), a cholinesterase inhibitor, physostigmine (0.003-0.1 mg/kg), and the quaternary derivative of physostigmine, neostigmine (0.003-0.1 mg/kg), were determined. Diazepam decreased matching accuracy such that a decrease in the mean delay value for the session was observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Cytokines help suggest aplastic anemia with pulmonary bacterial or co-fungal infection.
Although aplastic anemia (AA) does not come under the category of blood malignant diseases, the infection that frequently occurs in this bone marrow failure can make it worse. Pulmonary infection is the most prevalent but limiting clinical diagnosis. To find biomarkers predicting bacterial or bacterial-combined fungal infections in the lungs, we reviewed 287 AA medical records including 151 without any infection, 87 with pure pulmonary bacterial infection, and 49 with bacterial and fungal infection were reviewed. There were substantial changes in IL-17F, IL-17A, IFN-γ, IL-6, IL-8, and IL-10 levels between the non-infected and lung bacterial infection groups (P < 0.05). Further, a significant variation in IL-17A, TNF-β, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-22, and IL-12p70, between the uninfected group and the pulmonary bacterial and fungal infection group (P < 0.05) was observed. The results further revealed significant differences in TNF-β, IL-12p70, IL-6, IL-8, and IL-10 between the pulmonary bacterial infection group and the fungal infection group (P < 0.05). Moreover, by calculating ROC and cut-off values, we determined that IL-6 (AUC = 0.98, Cut-off = 14.28 pg/ml, P = 0.0000) had a significant advantage than other cytokines, body temperature (AUC = 0.61, P = 0.0050), PCT (AUC = 0.57, P = 0.0592), and CRP (AUC = 0.60, P = 0.0147) in the detection of lungs bacterial infections. In addition, IL-6 (AUC = 1.00, Cut-off = 51.50 pg/ml, P = 0.000) and IL-8 (AUC = 0.87, Cut-off = 60.53 pg/ml, P = 0.0000) showed stronger advantages than other cytokines, body temperature (AUC = 0.60, P = 0.0324), PCT (AUC = 0.72, Cut-off = 0.63 ng/ml, P = 0.0000) and CRP (AUC = 0.79, Cut-off = 5.79 mg/l, P = 0.0000) in distinguishing bacteria from fungi. This may suggest that IL-8 may play a role in differentiating co-infected bacteria and fungi. Such advantages are repeated in severe aplastic anemia (SAA) and very severe aplastic anemia (VSAA).In conclusion, aberrant IL-6 elevations in AA patients may predict the likelihood of bacterial lung infection. The concurrent increase of IL-6 and IL-8, on the other hand, should signal bacterial and fungal infections in patients.These findings may help to suggest bacterial or fungal co-infection in patients with AA (Focus on VSAA and SAA).


rbcL-based dataset on intra-specific diversity and conservation of Adansonia digitata L. (Malvaceae) in the savannah belt of Nigeria.
The African baobab species belong to the family Malvaceae and the genus Adansonia. The disjointed tree thrives in arid or semi-arid regions, is native to the thorn woodlands of Africa, along tracks, and is associated with human-populated forest areas. It is considered indigenous to Central and West Africa and has been introduced to the Arabian Peninsula, South-East Asia, the Indian sub-continent, and the Caribbeans. Adansonia digitata is a multifunctional tree with a long lifespan of over 1000 years old. The leaves, roots, flowers, fruit pulp, seeds and barks are used for food, medicine, or other ethnic-practices. The utilisation level and distribution are significantly undermined by climate change and poor use practices. The data set offers insight into the distribution pattern and genetic diversity of Adansonia digitata across the savannah belt of Nigeria using the rbcL gene.


Prevalence and patterns of caries, rampant caries, and oral health in two- to five-year-old children in Saudi Arabia.
The aim of this study was to determine the prevalence, severity, and pattern of dental caries, as well as the prevalence of rampant caries in preschool children in Jeddah, Saudi Arabia and to relate this to socioeconomic factors, diet, and oral hygiene practices. A random sample of 987 children was drawn from 17 nursery schools. Caries were diagnosed in 720 (73%) of the children: 256 (26%) of the children had caries confined to posterior teeth, 54 (6%) had caries only in anterior teeth, and in 382 (39%) both tooth types were affected. Forty-eight children (5%) had caries only in incisors. Rampant caries was diagnosed in 336 (34%). Mean dmft for the 987 children was 4.80 (+/- 4.87) per child and mean dmfs 12.67 (+/- 15.46). Caries and rampant caries were related to social class. There was little evidence of differing patterns of caries having clearly defined etiology.


PEDV ORF3 encodes an ion channel protein and regulates virus production.
Several studies suggest that the open reading frame 3 (ORF3) gene of porcine epidemic diarrhea virus (PEDV) is related to viral infectivity and pathogenicity, but its function remains unknown. Here, we propose a structure model of the ORF3 protein consisting of four TM domains and forming a tetrameric assembly. ORF3 protein can be detected in PEDV-infected cells and it functions as an ion channel in both Xenopus laevis oocytes and yeast. Mutation analysis showed that Tyr170 in TM4 is important for potassium channel activity. Furthermore, viral production is reduced in infected Vero cells when ORF3 gene is silenced by siRNA. Interestingly, the ORF3 gene from an attenuated PEDV encodes a truncated protein with 49 nucleotide deletions, which lacks the ion channel activity.


Effect of chorionic gonadotropin, triamcinolone, progesterone and estrogen on enzymes of placenta and liver in rats.
The activity of several enzymes of regulatory importance for the pathways of glycolysis, gluconeogenesis and lipogenesis was investigated in the placenta and liver of pregnant rats and in the liver of non-pregnant female rats. The rats received daily hormonal treatments on Days 15 to 17 of pregnancy and enzyme activities were measured on Day 18. Chorionic gonadotropin induced minor changes in enzyme activity, apart from a decrease in the activity of hepatic enzymes of lipogenesis in non-pregnant rats. Triamcinolone induced a marked increase in enzymes of gluconeogenesis and a decrease in the activity of pyruvate kinase in the liver of pregnant and non-pregnant rats; in contrast, inverse changes in activity, these enzymes were observed in the placenta. This response in the placenta was considered to arise not from direct hormone effect, but from the accompanying hyperglycemia and hyperinsulinemia. Triamcinolone also increased the activity of hepatic acetyl-CoA carboxylase in pregnant and non-pregnant rats, whereas it reduced the activity of this enzyme in the placenta. Estrogen produced changes similar to those of triamcinolone in the liver and placenta, except that it depressed the activity of acetyl-CoA carboxylase in both tissues. Progesterone had little effect on placental and hepatic enzymes. In general, the changes induced by these hormones in the placenta affected fewer enzymes than in the liver, were less extensive in magnitude and not necessarily in the same direction as in the liver. This indicates that the regulatory placental enzymes are subject to specific control mechanisms not necessarily influenced by direct hormone action.


[Quipazine antagonizing baclofen activity. Influence of baclofen and gamma-hydroxybutyric acid on serotonin metabolism in rat brain (author's transl)].
Baclofen increases the concentration of serotonin (5-HT) and 5-hydroxyindolacetic acid (5-HIAA) in the rat c. striatum without causing concomitant increase in 5-HT biosynthesis. The effect of baclofen on 5-HIAA, but not that on 5-HT, was antagonized dose-dependently by quipazine. It is assumed that quipazine acts by stimulating 5-HT autoreceptors.


Changes in polyphenolics during maturation of Java plum (Syzygium cumini Lam.).
Java plum (Syzygium cumini Lam.) is a rich source of polyphenolics with many purported health benefits, but the effect of maturation on polyphenolic content is unknown. Freeze-dried samples of Java plum from seven different maturity stages were analyzed for anthocyanin, flavonol, flavanonol and hydrolysable tannin composition by HPLC. Anthocyanins were first detected at the green-pink stage of maturity and increased throughout maturation with the largest increase occurring from the dark purple to black stages of maturation. Levels of gallotannins, ellagitannins, flavonols, gallic acid and ellagic acid were highest at early stages of maturation and decreased as the fruit ripened. For production of antioxidant-rich nutraceutical ingredients, fruit should be harvested immature to obtain extracts rich in hydrolysable tannins and flavonols. The exceptional anthocyanin content of black fruit may prove useful as a source of a natural colorant.


Costs and insurance coverage associated with permanent mechanical cardiac assist/replacement devices in the United States.
Each year over 50,000 persons in the United States could potentially benefit from some form of permanent cardiac replacement or assistance. Approximately 7000 of these persons get on the waiting list for a transplant, and 2300 are transplanted. About 2000 patients are reportedly exposed to a mechanical cardiac assist device, most often as a bridge to transplant. The majority of persons who might benefit from cardiac replacement are never referred for treatment and, thus, the number of deaths on the waiting list is a misleading indicator of access to transplantation and overall patient mortality. The total economic burden associated with coronary artery disease and congestive heart failure now exceeds $140 billion each year, with approximately $700 million directly spent on heart transplant procedures alone. If a viable total artificial heart is devised to replace a failed heart, or a ventricular assist system to permanently assist a failing heart, direct aggregate expenditures alone are likely to be somewhere between $5.4 and $24.0 billion annually. Based on individual patient care costs, as well as aggregate national expenditures, insurers will be reluctant to pay for the permanent use of such devices, even though cost is reportedly not a consideration in coverage decisions. Today, medical benefits and added value are concepts that will shape the coverage determination process, as will increasingly liberal policies regarding payment for treatment costs in relationship to clinical trials. Nonetheless, resource allocation and rationing decisions loom large as strange "characters at play" on an international economic "stage," while being "directed" by worldwide health care needs.


Inflammatory response after out-of-hospital cardiac arrest-Impact on outcome and organ failure development.
Post-cardiac arrest syndrome that occurs in out-of-hospital cardiac arrest (OHCA) patients is characterized by inflammatory response. We conducted a scoping review of current evidence regarding several inflammatory markers' usefulness for assessment of patient outcome and illness severity. We also discuss the proposed underlying mechanisms leading to inflammatory response after OHCA. We searched the MEDLINE, PubMed Central, Cochrane CENTRAL and Web of Science Core Collection databases with the following search terms: ("inflammation" OR "cytokines") AND "out-of-hospital cardiac arrest." Each inflammatory marker found was combined with "out-of-hospital cardiac arrest" using "AND" to find further relevant studies. We included original studies measuring inflammatory markers in adult OHCA patients that assessed their prognostic capabilities for mortality, neurological outcome, or organ failure severity. Fifty-nine studies met the inclusion criteria, covering in total 65 different markers. Interleukin-6 (IL-6), procalcitonin (PCT) and C-reactive protein (CRP) were the most studied markers, and they were associated with poor outcomes in 13/15, 13/14 and 11/17 studies, respectively. Based on area under the receiver operating characteristic curve (AUC) value, the time point of best discriminatory capacity for poor outcome was ICU admission for IL-6 (median AUC 0.78, range 0.71-0.98) and day one after OHCA for PCT (median AUC 0.84, range 0.61-0.98). Seven studies reported AUCs for CRP (range 0.52-0.76) with no measurement time point being superior to others. The association of IL-6 and PCT with outcome appeared stronger in studies with more severely ill patients. Studies reported conflicting results regarding each marker's association with organ failure severity. Inflammatory markers are potentially useful for early risk stratification after OHCA. PCT and IL-6 have moderate prognostic value during the first 24 h of the ICU stay. Predictive accuracy appears to be associated with the study overall event rate.


Immunostaining evidence for PI(4,5)P2 localization at the leading edge of chemoattractant-stimulated HL-60 cells.
It is well known that in fMLP-stimulated neutrophils, phosphatidyl inositol 3,4,5-trisphosphate [PI(3,4,5)P3] localizes at the leading edge of the cells. However, no effort has been made to study the PI 4,5-bisphosphate [PI(4,5)P2] distribution in these cells. In fact, it has been suggested that PI(4,5)P2 is unlikely to localize, as its basal level is orders of magnitude higher than that of PI(3,4,5)P3. We developed an optimized immunostaining protocol for studying the endogenous distribution of PI(4,5)P2 in neutrophil-like HL-60 cells. We show that PI(4,5)P2 localizes sharply at the leading edge with an intensity gradient similar to that for PI(3,4,5)P3. The enzymes for the production of PI(4,5)P2, namely, PI5KIalpha and PI5KIgamma, were also found to localize at the leading edge, further supporting our finding that PI(4,5)P2 localizes at the leading edge. These results imply that complementary regulation of PI3K and phosphate and tensin homolog (PTEN) is not the sole or dominant mechanism of PI(3,4,5)P3 polarization in HL-60 cells.


Correction: High expression of HMGA2 independently predicts poor clinical outcomes in acute myeloid leukemia.
Since the publication of the original article the authors noticed the the affiliation details for Paresh Vyas are incorrect. The correct affiliation details for this author are given below.


Intracellular mechanisms of cytoplasmic Ca2+ oscillation in rat megakaryocyte.
Extracellular application of ATP and ADP evoked the oscillatory K+ currents (IKCa) resulting from the periodic rise in cytoplasmic Ca2+ concentration ([Ca2+]i) of megakaryocyte isolated from rat bone marrow (Uneyama, H., Uneyama, C., and Akaike, N. (1992) Jpn. J. Pharmacol. 58, 231). The intracellular mechanism of ATP-induced cytoplasmic Ca2+ oscillation was investigated by the use of nystatin-perforated patch-clamp technique. Caffeine and ryanodine, which release Ca2+ from the Ca(2+)-induced Ca2+ release pool (CICR), and procaine, a blocker of Ca2+ release from CICR, had no effect on the IKCa oscillation in megakaryocyte. Thapsigargin and A23187 activated IKCa irreversibly by mobilizing Ca2+, but under Ca(2+)-free conditions they activated IKCa transiently. Intracellular application of inositol 1,4,5-triphosphate (IP3) also induced IKCa oscillation. Phorbol myristate acetate (PMA), a strong activator of protein kinase C (PKC), inhibited the oscillation completely. The inhibitory action of PMA was reversed by an inhibitor of PKC, staurosporin. The oscillation of ATP-induced IKCa was disrupted by staurosporin or calmodulin (CaM) antagonists such as W-7 and trifluoperazine, resulting in a transient and successive plateau-like IKCa. These results suggest that Ca2+ oscillation in megakaryocyte is caused by the interaction of both the Ca2+ release from IP3-sensitive Ca2+ pool and the Ca2+ uptake stimulated by PKC and Ca2+/CaM complex. Furthermore, forskolin, an activator of adenylate cyclase, and isobutylmethylxanthin, an inhibitor of phosphodiesterase, inhibited the frequency, latency, and current amplitude of the oscillation in a concentration-dependent manner. These reagents inhibited IP3-induced oscillation as well as an ATP-induced oscillation. Thus, the ATP-induced [Ca2+]i oscillation of rat megakaryocyte is also modulated by cAMP.


Halothane solubility in blood during cardiopulmonary bypass: the effect of haemodilution and hypothermia.
The influence of haemodilution and cooling on the solubility of halothane in blood during cardiopulmonary bypass was studied in six normothermic and six hypothermic patients undergoing open-heart operations. The solubility of halothane in blood was 2.17 at 30 degrees C and 1.34 at 37 degrees C. There was also an increase in the blood/gas partition coefficient of about 8.7 per cent for each degree celsius of temperature fall. These changes in haemodiluted blood explain the different anaesthetic requirements observed in patients undergoing cardiopulmonary bypass, either at normal temperature or at moderate hypothermia.


Resolution of acute intestinal graft-versus-host disease.
Allogeneic transplantation of hematopoietic stem cells (allo-HCT) represents an increasingly employed therapeutic approach to potentially cure patients suffering from life-threatening malignant and autoimmune disorders. Despite its lifesaving potential, immune-mediated allo-reactivity inherent to the allogeneic transplantation can be observed within up to 50% of all allo-HCT patients regularly resulting in the manifestation of acute and/or chronic graft-versus-host disease (GvHD). Mechanistically, especially donor T cells are assumed to chiefly drive inflammation that can occur in virtually all organs, with the skin, liver, and gut representing as the most frequently affected anatomic sites. Especially in the presence of intestinal manifestations of GvHD, the risk that the disease takes a life-threatening, potentially fatal course is significantly increased. In the light of a rapid gain of knowledge in respect to decode innate and adaptive immunity related mechanisms as, e.g., cytokine networks, intracellular signaling pathways or environmental triggers as, e.g., the intestinal microbiota and the development of novel therapeutic approaches, detailed insight into endogenous mechanisms seeking to counterbalance the proinflammatory machinery or to proactively foster signals promoting the resolution of allo-driven intestinal inflammation is emerging. Here, we seek to highlight the key aspects of those mechanisms involved in and contributing to the resolution of GvHD-associated intestinal inflammation. Concomitantly, we would like to briefly outline and discuss promising future experimental targets suitable to be therapeutically employed to directionally deflect the tissue response from a proinflammatory to an inflammation-resolving type of intestinal GvHD after allo-HCT.


The treatment of pelvic inflammatory disease.
The treatment of pelvic inflammatory disease depends upon the etiology of the condition. Pelvic infection (PI) after parturition and abortion, gynecologic surgery, and a variety of invasive procedures is commonly associated with the isolation of anaerobic and aerobic flora of the vagina. The factors which influence the choice of antimicrobial treatment and the role of Bacteroides fragilis and Escherichia coli are discussed. Sexually transmissible agents of importance are Neisseria gonorrhoeae and Chlamydia trachomatis. Pelvic infections associated with these pathogens require antibiotics which exert an optimum effect against them. Examination and treatment of the sexual partner(s) are important. The possible role of the anaerobic and aerobic vaginal flora as opportunistic secondary pathogens is discussed. Developments in the surgical treatment of the sequelae of PID are reviewed. The results of treatment of uncomplicated gonorrhea in 262 women are reported. C. trachomatis was isolated from 53% of women before treatment. After treatment, PI developed in 11 women who had been given penicillin and in one woman who had been given tetracycline (P = 0.0071). It is suggested that recognition and treatment of postgonococcal cervicitis in women treated for uncomplicated gonorrhea with penicillin might provide one form of preventive treatment for nongonococcal PI.


The current Monkeypox outbreak: Updates and concerns.
Since 1958 witnessed the detection of Monkeypox virus in monkeys, no human infection was encountered until 1970. Afterwards, zoonotic transmission was the rule near African rainforests, mainly in DRC. Most cases occurred in children who weren't immunized against smallpox. Since 2003 and the first human infection in the USA, research was accelerated. Two clades were identified with different virulence, demographic distribution and transmissibility. The mean age of infection increased with waning smallpox vaccine immunity. Mild febrile prodrome can precede lymphadenopathy, which doesn't occur in smallpox. Homogenous crops of lesions appear in stages until scabs fall and contagiosity ends. However, since May outbreak, cases started to appear in non-endemic areas, human transmission increased and was linked to close sexual contact especially in MSM community. Lesions were found mainly perioral, at genitals and perianal. Newer system for nomenclature was suggested in which there are 3 viral clades and the responsible clade for the outbreak is clade 3 (lineage B.1). About 50 mutations were detected compared with the strains isolated 4 years ago. Gene loss and APOBEC3 may be related to accelerated mutation rate which may accelerate human transmission. Previous mistakes in failure to allocate available vaccines to control the disease in previously endemic areas should be avoided and rapid ring vaccination of potential contacts and those at risk should be a priority. Case isolation, contact isolation or tracing for an incubation period, standard measures for airborne infections and safe sex should be implanted in the light of the current uncertainty.


Intrinsically distinct hole and electron transport in conjugated polymers controlled by intra and intermolecular interactions.
It is still a matter of controversy whether the relative difference in hole and electron transport in solution-processed organic semiconductors is either due to intrinsic properties linked to chemical and solid-state structure or to extrinsic factors, as device architecture. We here isolate the intrinsic factors affecting either electron or hole transport within the same film microstructure of a model copolymer semiconductor. Relatively, holes predominantly bleach inter-chain interactions with H-type electronic coupling character, while electrons' relaxation more strongly involves intra-chain interactions with J-type character. Holes and electrons mobility correlates with the presence of a charge transfer state, while their ratio is a function of the relative content of intra- and inter-molecular interactions. Such fundamental observation, revealing the specific role of the ground-state intra- and inter-molecular coupling in selectively assisting charge transport, allows predicting a more favorable hole or electron transport already from screening the polymer film ground state optical properties.


Three galactose inducible promoters for use in C. neoformans var. grubii.
Cryptococcus neoformans is the causative agent of cryptococcal meningoencephalitis, most frequently occurring in immunocompromised individuals. There are three varieties of C. neoformans, var. grubii, var. neoformans, and var. gatti. Worldwide var. grubii is the most prevalent clinical isolate. However, few tools for the study of essential genes in var. grubii exist. Here we describe three endogenous inducible promoters for use in the study of this important opportunistic pathogen. We identified eight potential homologs of S. cerevisiae galactose genes in var. grubii. We found that GAL1, GAL7, and UGE2 were regulated by glucose and galactose and can be used successfully during mating. Our analysis indicated these promoters should prove to be excellent tools for analysis of genes in var. grubii.


Completeness of case ascertainment at the Irish National Cancer Registry.
Cancer registries play a major role in cancer control, including quantifying the burden of cancer in the population, informing health service planning, and evaluating service provision. Registries must achieve a high level of completeness of case ascertainment. Completeness of case ascertainment for the National Cancer Registry (NCR) in Ireland was estimated. For all invasive cancers, and the four most common sites, we used two quantitative methods which use information from death certificates to obtain estimates of completeness of registration-the Lincoln-Petersen (LP) estimator (which provides a lower bound estimate) and the flow method (which estimates completeness at a fixed time-point, in this case 5 years). Data were extracted at the end of 2010. Independent case ascertainment was used to assess completeness of breast cancer registration in the 50-64 age group and of rectal cancer in surgical patients diagnosed in 2007. Completeness of case ascertainment was >99 % for the breast screening age-group and for surgical rectal cancer cases. A lower bound on completeness of case ascertainment for all invasive cancers excluding non-melanoma skin cancers is 94.3 % using the LP estimator. The estimate from the flow method is 97 % after 5 years, [95 % confidence interval (96.2 %, 97.7 %)]. Five-year levels of completeness for the four commonest cancers using the flow method were: colorectal, 97.4 %; lung, 98.7 %; prostate, 95.8 % and breast, 98.0 %. The estimated levels of completeness for the NCR are satisfactory. Further work is required to compare the methods used.


A deactivating conformational change induced by reduced nicotinamide-adenine dinucleotide phosphate in a Neurospora glutamate dehydrogenase.
Stopped-flow fluorescence techniques have been used to observe the formation of the binary comples of E-NADPH. At pH 7.5 there is a protein conformational change after the formation of the binary complex. This conformational change can be detected by a decrease in the fluorescence intensity of the complex at 350 nm and by an increase in its fluorescence intensity at 450 nm.


Immunohistochemical studies of the carbonic anhydrase isozymes in the bovine placenta.
Placentae from 11 cows, ranging from about 80 to 270 days gestation, were studied for immunohistochemical localization of carbonic anhydrase isozymes. CA isozymes were localized using the avidin-biotin-peroxidase complex (ABC) method in the bovine placenta. CA-II was found in the fetal trophoblastic cells with single nuclei as well as in erythrocytes of maternal and fetal blood. The percentage of positive cells for anti-CA-II in the trophoblastic cells did not change during development of gestation in the fetal bovine. CA-I and CA-III were not detected in the bovine placenta.


Comprehending the impossible: what role do selectional restriction violations play?
To elucidate how different kinds of knowledge are used during comprehension, readers' eye movements were monitored as they read sentences that were: plausible, impossible because of a selectional restriction violation, or impossible because of a violation of general world knowledge. Eye movements on the pre-critical, critical, and post-critical words evidenced disruption in the selectional restriction violation condition compared to the other two conditions. These findings suggest that disruption associated with reading about impossible events is not directly determined by how impossible the event seems. Rather, the relationship between the verb and arguments in the sentence seems to matter. These findings are the strongest evidence to date that processing effects associated with selectional restrictions can dissociate from those associated with general world knowledge about events.


The Effect of Trimethoprim on Thiamine Absorption: A Transporter-Mediated Drug-Nutrient Interaction.
Trimethoprim is predicted to inhibit several thiamine transporters, including the primary thiamine intestinal absorptive transporter, ThTR-2, and the hepatic and renal organic cation transporters, OCT1, OCT2, and MATEs. To investigate the effect of trimethoprim on thiamine absorption, studies were conducted in cells, mice, and healthy volunteers and supported by use of real-world data. In a randomized, crossover clinical study, seven healthy volunteers were given a single oral dose of thiamine or thiamine plus trimethoprim, followed by blood sampling. The thiamine area under the curve (AUC) increased with trimethoprim co-administration (P value = 0.031). Similar results were seen in mice. Trimethoprim appeared to act on thiamine absorption through inhibition of hepatic OCT1 as evidenced from its ability to modulate levels of isobutyrylcarnitine and propionylcarnitine, OCT1 biomarkers identified from metabolomic analyses. Real-world data further supported this finding, showing an association between trimethoprim use and higher levels of triglycerides, LDL cholesterol, and total cholesterol, consistent with OCT1 inhibition (P values: 2.2 × 10-16 , 5.75 × 10-7 , and 5.82 × 10-7 , respectively). These findings suggest that trimethoprim increases plasma levels of thiamine by inhibiting hepatic OCT1. Trimethoprim reduced urinary excretion and clearance of biomarkers for OCT2 and MATEs, consistent with inhibition of renal organic cation transporters. This inhibition did not appear to play a role in the observed increases in thiamine levels. This study highlights the potential for drug-nutrient interactions involving transporters, in addition to transporters' established role in drug-drug interactions.


Global hippocampus functional connectivity as a predictive neural marker for conversion to future mood disorder in unaffected offspring of bipolar disorder parents.
Hippocampus-related functional alteration in genetically at-risk individuals may reflect an endophenotype of a mood disorder. Herein, we performed a prospective study to investigate whether baseline hippocampus functional connectivity (FC) in offspring of patients with bipolar disorder (BD) would predict subsequent conversion to mood disorder. Eighty bipolar offspring and 40 matched normal controls (NC) underwent resting state functional MRI (rsfMRI) scanning on a 3.0 Tesla MR scanner. The offspring were subdivided into asymptomatic offspring (AO) (n = 41) and symptomatic offspring (SO) (n = 39) according to whether they manifested subthreshold mood symptoms. After identifying the different hippocampus FCs between the AO and SO, a logistic regression analysis was conducted to investigate whether the baseline hippocampus FCs predicted a future mood disorder during a 6-year follow-up. We identified seven baseline para/hippocampus FCs that showed differences between AO and SO, which were entered as predictive features in the logistic regressive model. Of the 80 bipolar offspring entering the analysis, the FCs between left hippocampus and left precuneus, and between right hippocampus and left posterior cingulate, showed a discriminative capacity for predicting future mood disorder (area-under-curve, or AUC=75.76 % and 75.00 % respectively), and for predicting BD onset (AUC=77.46 % and 81.63 %, respectively). The present findings revealed high predictive utility of the hippocampus resting state FCs for future mood disorder and BD onset in individuals at familial risk. These neural markers can potentially improve early detection of individuals carrying particularly high risk for future mood disorder.


Zidovudine disposition in patients with severe renal impairment: influence of hemodialysis.
Pharmacokinetics of zidovudine (azidothymidine, AZT) was investigated after oral administration (200 mg) in 25 HIV seronegative subjects: 14 patients with severe renal impairment (creatinine clearance 6 to 31 ml/min), five hemodialyzed anuric patients, and six healthy subjects. Plasma and urine concentrations of zidovudine and its glucuronidated metabolite (GAZT) were measured simultaneously by HPLC assay. In healthy subjects, GAZT concentrations were higher than those of AZT; AUC values were 23.7 +/- 1.9 and 5.2 +/- 0.6 mumol.hr/L, respectively. Formation of GAZT rate-limits its elimination: GAZT half-life (t 1/2) parallels that of AZT, which is around 1 hour. In uremic patients, AZT concentrations were moderately increased (AUC = 11.7 +/- 1.1 mumol.hr/L), whereas t 1/2 and mean residence time (MRT) remain unchanged despite the decreased renal clearance (16 +/- 2 versus 220 +/- 58 ml/min) and decreased urinary excretion (1.6 +/- 0.3 versus 8.1 +/- 1.0% of the dose). In contrast, GAZT concentrations are markedly increased (AUC = 402.9 +/- 88.6 mumol.hr/L). As a consequence of the decreased renal clearance (27 +/- 3 versus 331 +/- 42 ml/min), elimination is the rate-limiting step and t 1/2 is increased (8 +/- 2 versus 0.9 +/- 0.1 hr). Contribution of a 4-hour hemodialysis session to AZT elimination appears to be negligible, whereas elimination of GAZT is enhanced. On the sole basis of AZT pharmacokinetic data, no particular dose adjustment appears to be necessary in patients who have severe renal impairment (creatinine clearance between 10 and 30 ml/min). However, high levels of GAZT should be anticipated with the usual dosage regimen.


Imputation of sequence level genotypes in the Franches-Montagnes horse breed.
A cost-effective strategy to increase the density of available markers within a population is to sequence a small proportion of the population and impute whole-genome sequence data for the remaining population. Increased densities of typed markers are advantageous for genome-wide association studies (GWAS) and genomic predictions. We obtained genotypes for 54 602 SNPs (single nucleotide polymorphisms) in 1077 Franches-Montagnes (FM) horses and Illumina paired-end whole-genome sequencing data for 30 FM horses and 14 Warmblood horses. After variant calling, the sequence-derived SNP genotypes (~13 million SNPs) were used for genotype imputation with the software programs Beagle, Impute2 and FImpute. The mean imputation accuracy of FM horses using Impute2 was 92.0%. Imputation accuracy using Beagle and FImpute was 74.3% and 77.2%, respectively. In addition, for Impute2 we determined the imputation accuracy of all individual horses in the validation population, which ranged from 85.7% to 99.8%. The subsequent inclusion of Warmblood sequence data further increased the correlation between true and imputed genotypes for most horses, especially for horses with a high level of admixture. The final imputation accuracy of the horses ranged from 91.2% to 99.5%. Using Impute2, the imputation accuracy was higher than 91% for all horses in the validation population, which indicates that direct imputation of 50k SNP-chip data to sequence level genotypes is feasible in the FM population. The individual imputation accuracy depended mainly on the applied software and the level of admixture.


A Cross-Sectional Geographic Information Systems Study of a Pediatric Emergency Department Child Restraint System Distribution Program.
A pediatric ED program sought to promote injury prevention through distribution of child restraint systems. Program funds are paid for child passenger safety technician certification of all personnel. Pediatric emergency nurses distributed child restraint systems at hospital discharge and dedicated technicians at fitting stations. Researchers described program characteristics, developed a baseline understanding of program outreach using geographic information systems, and evaluated adherence to manufacturer guidelines with a sensitivity analysis. This retrospective cross-sectional study used distribution forms linked to hospital records from 2013 to 2016. Testing for differences used nonparametric methods. Median values and interquartile ranges for weight and height of children were compared with manufacturer guidelines. Geographic information systems visualized recipients' street addresses and motor vehicle crashes on an underlying base map. There were 312 child restraint systems distributed: of which 179 (57.4%) at the hospital, 126 (40.4%) at fitting stations, and 7 (2.2%) missing a location. Among those on Medicaid, 64.4% received a child restraint system at the hospital compared with 35.6% at fitting stations (χ2 = 5.40, P < .02). Fitting stations had limited outreach to rural residents. Finally, results from the sensitivity analysis showed that devices were issued according to manufacturer guidelines. Despite the workplace pressures of clinical care, pediatric emergency nurses delivered educational information and demonstrated hands-on installation at similar rates to dedicated technicians. Distribution of child restraint systems through the hospital reached a uniquely underserved population. Further research should investigate methods to improve fitting station outreach among Medicaid recipients.


Strain differences in proliferation of progenitor cells in the dentate gyrus of the adult rat and the response to fluoxetine are dependent on corticosterone.
This paper investigates the role of differences in adrenal cortical function on the proliferation rate of progenitor cells in the dentate gyrus of the hippocampus in adult Sprague-Dawley (SD) and Lister-Hooded (LH) male rats. SD rats had around 60% more cells labeled with Ki67 (an index of mitosis) than LH rats under basal conditions. Bilateral adrenalectomy (ADX) increased levels in both strains, but by unequal amounts, such that post-ADX numbers of Ki67-labeled cells were similar in both strains. Daily injections of 5 mg/kg corticosterone for 7 days reduced levels to similar values in ADX rats of both strains. The activity of progenitor cells in either strain did not respond to daily i.p. injections of fluoxetine (10 mg/kg) for 14 days, but an equivalent dose administered by osmotic minipump stimulated proliferation in both by a similar proportional amount, such that strain differences persisted. S.c. implantation of a corticosterone pellet (75 mg), which flattens the diurnal rhythm in corticosterone, prevented fluoxetine delivered by minipump from activating progenitor cell mitosis in SD rats, as it had in the LH strain in a previous study. These results show that much, if not all, of the marked strain differences between SD and LH rats in progenitor cell activity, and hence rates of neurogenesis in the dentate gyrus may be ascribed to corresponding differences in adrenal cortical activity.


Predicting malignancy in thyroid nodules with benign cytology results: The role of Conventional Ultrasound, Shear Wave Elastography and BRAF V600E.
Ultrasound-guided fine-needle aspiration (US-FNA) is the most accurate method for preoperative diagnosis of thyroid nodules, but how to deal with false negative results? This study aimed to find preoperative diagnosis methods including Conventional Ultrasound (CUS), Shear Wave Elastography (SWE) and BRAF V600E testing to differentiate false negative nodules. Forty-nine nodules in 49 patients with benign FNA results and pathological diagnoses were included. CUS and SWE features were evaluated. BRAF V600E analysis was performed after FNA. Diagnostic performances of three methods were analyzed in predicting malignancy in benign FNA results. Twenty-seven of 49 nodules were malignant, and 22 nodules were benign. Hypoechogenicity, taller-than-wider, irregular boundary, microcalcification, SWE max, SWE mean and BRAF V600E mutation were risk factors for malignancy. All 7 malignant nodules with BRAF V600E mutations and 18 of 20 malignant nodules without BRAF V600E mutations have two or more suspicious CUS features. Six of 7 malignant nodules with BRAF V600E mutations and 16 of 20 malignant nodules without BRAF V600E mutations had SWE mean value greater than the cut-off value. CUS, SWE and BRAF V600E were diagnostic tools for malignancy in FNA benign nodules. Further clinical decisions should be considered for nodules with two or more suspicious CUS features and SWE parameters greater than cut-off values whether BRAF V600E is mutational or not.


Prevalence and correlates of pain and associated depression among community-dwelling older adults: Cross-sectional findings from LASI, 2017-2018.
Health practitioners often meet older persons suffering both from pain and depression. The study aimed to examine the factors associated with pain among older individuals and its association with major depression. In addition, the interaction between self-rated health (SRH), wealth status, and pain that is associated with depression are explored. We used data from the Longitudinal Aging Study in India (LASI). Participants included 15,098 male and 16,366 female adults aged 60 years or older. Univariate and bivariate analyses along with χ2 tests were conducted in the initial stage. Binary logistic regression analyses were performed to fulfill the objectives. Major depression was calculated using Short Form Composite International Diagnostic Interview. A proportion of 39.6% older individuals (n = 12,686) reported pain and 8.7% older adults (n = 2657) suffered from depression. Older adults who suffered from pain frequently were more likely to be depressed than those who never suffered from pain (adjusted odds ratio [AOR]: 1.70; confidence interval [CI]: 1.38-2.09). The interaction of pain, SRH, and household wealth status on depression found that older adults with pain who reported poor SRH (AOR: 4.18; CI: 3.50-5.00) or belonged to rich households (AOR: 2.27; CI: 1.84-2.80) had higher odds of suffering from depression, compared to older adults with no pain and good SRH or no pain and belonged to poor households. It is highlighted that pain is quite common in older people, and is linked to depression especially among older people with poor SRH. Thus, routine evaluation of pain and associated symptoms of mental illnesses should be performed for ensuring healthy aging.


Hexose metabolism in pancreatic islets. Metabolic and secretory responses to D-fructose.
D-Fructose (3.3 to 33.0 mmol/liter) caused a concentration-related increase in insulin output from rat islets exposed to D-glucose (3.3 to 7.0 mmol/liter), such an increase not being more marked in mouse islets. The fructose-induced increment in insulin release, relative to that evoked by D-glucose, was two times higher in islets exposed to D-glucose than in islets stimulated by D-mannose, 2-ketoisocaproate, or nonnutrient secretagogs. Likewise, the metabolism of D-fructose in islet cells was significantly different in the absence or presence of D-glucose. Thus, the ketose was largely channeled into the pentose phosphate pathway in glucose-deprived, but not so in glucose-stimulated, islets. In both glucose-deprived and glucose-stimulated islets, however, the magnitude of the secretory response to D-fructose was commensurate with the increase in ATP production attributable to its catabolism. These findings indicate that the metabolic fate of hexoses--and, hence, their insulinotropic capacity--is not ruled solely at the level of their phosphorylation.


Characterization of the pH-mediated solubility of Bacillus thuringiensis var. san diego native delta-endotoxin crystals.
Native crystals of Bacillus thuringiensis var. san diego, a coleopteran-specific delta-endotoxin, were metabolically labelled with [35S]methionine. Specific activity was 82,000 CPM/micrograms (2.44 Ci/mmol). Using a universal buffer formulated with the same ionic strength at every pH, we determined that native crystals dissolve above pH 10 and below pH 4. At the acidic pH, the rate of solubilization was substantially slower than at the alkaline pH. Recrystallization rates for the toxin were similar regardless of solubilization conditions. The banding patterns in denatured polyacrylamide gel electrophoresis were unaffected by solubilization conditions. Toxicity was higher for soluble toxin compared to crystal toxin, but virtually identical for the acidic and alkaline produced solutions. Acid solubilization is significant because of the acidic midgut of susceptible Coleoptera.


The case of the submandibular gland sialolith.
The authors describe a case of a sialolith formation in a submandibular gland over 2 years. A large salivary stone is demonstrated radiologically and surgical treatment is described.


Elevated levels of a U4/U6.U5 snRNP-associated protein, Spp381p, rescue a mutant defective in spliceosome maturation.
U4 snRNA release from the spliceosome occurs through an essential but ill-defined Prp38p-dependent step. Here we report the results of a dosage suppressor screen to identify genes that contribute to PRP38 function. Elevated expression of a previously uncharacterized gene, SPP381, efficiently suppresses the growth and splicing defects of a temperature-sensitive (Ts) mutant prp38-1. This suppression is specific in that enhanced SPP381 expression does not alter the abundance of intronless RNA transcripts or suppress the Ts phenotypes of other prp mutants. Since SPP381 does not suppress a prp38::LEU2 null allele, it is clear that Spp381p assists Prp38p in splicing but does not substitute for it. Yeast SPP381 disruptants are severely growth impaired and accumulate unspliced pre-mRNA. Immune precipitation studies show that, like Prp38p, Spp381p is present in the U4/U6.U5 tri-snRNP particle. Two-hybrid analyses support the view that the carboxyl half of Spp381p directly interacts with the Prp38p protein. A putative PEST proteolysis domain within Spp381p is dispensable for the Spp381p-Prp38p interaction and for prp38-1 suppression but contributes to Spp381p function in splicing. Curiously, in vitro, Spp381p may not be needed for the chemistry of pre-mRNA splicing. Based on the in vivo and in vitro results presented here, we propose that two small acidic proteins without obvious RNA binding domains, Spp381p and Prp38p, act in concert to promote U4/U5.U6 tri-snRNP function in the spliceosome cycle.


Air pollution and health risks due to vehicle traffic.
Traffic congestion increases vehicle emissions and degrades ambient air quality, and recent studies have shown excess morbidity and mortality for drivers, commuters and individuals living near major roadways. Presently, our understanding of the air pollution impacts from congestion on roads is very limited. This study demonstrates an approach to characterize risks of traffic for on- and near-road populations. Simulation modeling was used to estimate on- and near-road NO2 concentrations and health risks for freeway and arterial scenarios attributable to traffic for different traffic volumes during rush hour periods. The modeling used emission factors from two different models (Comprehensive Modal Emissions Model and Motor Vehicle Emissions Factor Model version 6.2), an empirical traffic speed-volume relationship, the California Line Source Dispersion Model, an empirical NO2-NOx relationship, estimated travel time changes during congestion, and concentration-response relationships from the literature, which give emergency doctor visits, hospital admissions and mortality attributed to NO2 exposure. An incremental analysis, which expresses the change in health risks for small increases in traffic volume, showed non-linear effects. For a freeway, "U" shaped trends of incremental risks were predicted for on-road populations, and incremental risks are flat at low traffic volumes for near-road populations. For an arterial road, incremental risks increased sharply for both on- and near-road populations as traffic increased. These patterns result from changes in emission factors, the NO2-NOx relationship, the travel delay for the on-road population, and the extended duration of rush hour for the near-road population. This study suggests that health risks from congestion are potentially significant, and that additional traffic can significantly increase risks, depending on the type of road and other factors. Further, evaluations of risk associated with congestion must consider travel time, the duration of rush-hour, congestion-specific emission estimates, and uncertainties.


A survey of the alpha-tubulin gene family in chicken: unexpected sequence heterogeneity in the polypeptides encoded by five expressed genes.
To characterize the alpha-tubulin gene family in chicken, we have isolated five chicken alpha-tubulin genes and determined the majority of the sequences of the encoded polypeptides. Three of these (c alpha 3, c alpha 5/6 and c alpha 8) encode novel, expressed alpha-tubulins that have not previously been analyzed, whereas one gene segment is a pseudogene and another appears capable of encoding a functional subunit (although we were unable to document its expression in a survey of chicken tissues). Together with two additional expressed, functional alpha-tubulins reported earlier, we conclude that the chicken alpha-tubulin family is comprised of at least five functional genes whose polypeptide products are substantially more heterogeneous than found in preceding analyses of vertebrate alpha-tubulins. Comparison of the amino acid sequences reveals that the five polypeptides are between 96 and 83% identical, with the extreme carboxy-terminal residues representing a highly heterogeneous variable domain. Since some alpha-tubulins undergo cyclic post-translational removal and readdition of a carboxy-terminal tyrosine, the notable sequence divergence in this domain suggests that individual tubulins probably participate to different extents in this modification cycle.


The TLR2 and TLR4 gene polymorphisms in Moroccan visceral leishmaniasis patients.
Visceral leishmaniasis (VL) is endemic in the Mediterranean basin and leads to the most severe form of Leishmania infection, lethal if left untreated. However, most infections are sub-clinical or asymptomatic, reflecting the influence of host genetic background on disease outcome. This study aimed to investigate possible association of TLR4 Asp299Gly, TLR4 Thr399Ile and TLR2 Arg753Gln polymorphisms with VL in Moroccan children. We enrolled 119 children with VL caused by Leishmania infantum as well as 138 unrelated children, 95 asymptomatic subjects and 43 healthy individuals who had no evidence of present or past infection. Polymorphisms were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system assay (ARMS-PCR). Results showed significant differences in genotype Thr399Ile and recessive model frequencies between VL and delayed-type hypersensitivity (DTH+) groups (p=0.018, OR=0.414CI 0.195-0.880; p=0.029, OR=0.448CI 0.214-0.938], respectively) by having the amino-acid threonine polymorphism as a reference in the VL group. Concerning the Asp299Gly there were a significant associations when comparing VL vs DTH+ (Asp299Gly genotype p=0.002, OR=0.326CI 0.158-0.671, allele frequencies p=0.033, OR=0.396CI 0.164-0.959, recessive model p=0.002, OR=0.343CI 0.172-0.681) and DTH+ vs DTH- groups (Asp299Gly genotype p=2.160E-4, OR=3.065CI 1.672-5.618, Gly299Gly genotype p=0.047, OR=0.368CI 0.299-0.452, allele frequencies p=1.406E-7, OR=29.571CI 3.907-223.8, recessive model p=4.370E-14, OR=36.965CI 8.629-158.3), by having the aspartic acid polymorphism as a reference these results suggest that the allele A (savage) confer protection against the clinical manifestations but not against the infection. Furthermore, there was a significant association regarding the Arg753Gln genotype (p=0.002, OR=0.326CI 0.158-0.671), allele frequencies (p=0.033, OR=0.396CI 0.164-0.959) and when applying a recessive model (p=0.002, OR=0.343CI 0.172-0.681) in the VL vs DTH+ groups. The same results was observed when comparing DTH+ vs DTH- groups (p=4.136E-6, OR=0.211CI 0.104-0.428), allele frequencies (p=0.008, OR=0.327CI 0.137-0.779) and recessive model (p=1.748E-5, OR=0.244CI 0.124-0.480). The results provide evidence that allele C in Thr399Ile and allele G in Arg753Gln polymorphisms may lead to protection against the clinical disease. Our data provide insights into the possible role of TLR2 and TLR4 variations in VL susceptibility.


Spectroscopic identification of the haem ligands of cellobiose oxidase.
A spectroscopic study of the flavocytochrome b enzyme, cellobiose oxidase, employing optical, NMR, EPR and near infra-red MCD techniques, has identified the axial ligands of the b-type haem. These are a histidine and a methionine, and this ligation set is discussed in relation to the functional role of the haem group.


Improvement of prognosis in breast cancer in Denmark 1977-2006, based on the nationwide reporting to the DBCG Registry.
Since 30 years DBCG (Danish Breast Cancer Coperative Group) has maintained, on a nation-wide basis, a clinical database of diagnostic procedures, therapeutic interventions, and clinical outcome in patients with primary breast cancer. The present analysis was undertaken to evaluate the development of the prognosis since 1977, and to analyse factors potentially contributing to the change of the prognosis. All cases of invasive breast cancer reported to DBCG during the period 1977-2006 were included in the present analysis. A total of close to 80 000 patients were registered in the DBCG Database. Since 1977 the prognosis has improved significantly, thus 5 year survival for the total population of patients with primary breast cancer has increased from 65 to 81%. According to the present analysis diagnosis at an earlier stage in the natural course of the disease and especially the development of more active systemic treatment modalities have contributed to the improved prognosis.


Effects of N-acetyl-L-cysteine on fish hepatoma cells treated with mercury chloride and ionizing radiation.
Organisms are exposed to natural radiations from cosmic or terrestrial origins. Furthermore the combined action of radiation with various chemicals is an inevitable feature of modern life. Radiation is known to cause cell death, mainly due to its ability to produce reactive oxygen species in cells. N-acetyl-L-cysteine (NAC) is a well-known sulfhydryl-containing antioxidant whose role in radioprotection has been reported. Synergistic effects of radiation and mercury chloride on human cells was previously reported by the authors. Based on the previous report, this study was designed to assess the synergistic effects of radiation and mercury chloride on fish hepatoma cells, as well as to investigate the protective effects of NAC on the cells. The cytotoxicity of radiation was enhanced in the presence of mercury chloride. NAC in lower concentrations prevented cells from death after irradiation with lower doses (<300 Gy) while it did not prevent cells from radiation-induced death after irradiation with higher doses (300, 500 Gy). The intracellular glutathione (GSH) levels significantly decreased after irradiation while the combined treatment of NAC and radiation alleviated the decrease in the GSH levels. The investigations give a clue for the action mechanism of synergistic or protective effects of NAC on the cells. Due to their high resistance to ionizing radiation, the PLHC-1 cells can be effectively used as a screening tool for assessing the combined effects of radiation with toxic chemicals.


[Paracentric inversions in man. Apropos of 2 familial observations].
Two observations of paracentric inversion in man are reported. One is located on the long arm of chromosome 1 and was observed over two generations. The other is on the long arm of chromosome 5 and was transmitted over three generations. The possible implications of paracentric inversions on the phenotype and fertility are discussed.


Epidermoid cysts of the liver.
To the four recorded cases of epidermoid cysts of the liver are added two cases recently seen in children. One was in a 4-yr-old girl and associated with biliary cirrhosis. The other was in a 5-mo-old boy. The former was treated by roux-en-y cystjejunostomy, the latter by enucleation. The origin of these cysts is unknown. One theory suggests derivation from accessory foregut buds. Because of their malignant potential, treatment should be by excision. Where this is impossible, roux-en-y cystjejunostomy offers satisfactory palliation.


Food chloride distribution in nature and its relation to sodium content.
Because of growing interest in the biological and clinical effects of dietary chloride as the anion accompanying the dietary cation sodium and because the standard food composition tables used in the United States to estimate sodium content do not contain data on chloride content, we analyzed the nutrient data base of the English workers Paul and Southgate, which contains an extensive listing of both chloride and sodium contents in foods. To examine food chloride distribution in nature, we focused on the uncooked, unadulterated, discrete, primitive foods in the data base (no. = 216 food items). The findings indicate the existence of both a large variability of chloride content among foods and a high degree of coupling of chloride with sodium. The contents of chloride and sodium varied over a similarly large range (coefficients of variation, 229% vs. 263%), differed very little from each other on the average (less than 20%), and correlated (r = 0.84, p less than 0.001) to the extent that greater than two-thirds of the overall variation of chloride content was linked to that of sodium content. Those findings accord with the often posited but untested assertion that the chloride content of foods approximates and parallels that of sodium.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of treatment with topical ivermectin three and eight weeks after turnout on nematode control and the performance of second-season beef suckler cattle.
Two groups of yearling suckled beef calves born between January and May of the preceding year and another two groups of lighter and slightly younger calves born between the previous March and July were grazed during their second year on four separate paddocks known to be contaminated with infective larvae of gastrointestinal nematodes. One of the heavier and one of the lighter groups were treated with a topical formulation of ivermectin three weeks and eight weeks after turnout. The treatment of the heavier group had only relatively minor parasitological effects and no effect on weight gains, whereas the treatment of the lighter cattle resulted in increased weight gains due to effective nematode control. The paper highlights that small differences in previous performances, age and exposure to parasites can have a substantial impact on the benefits accrued from anthelmintic treatment.


A comparison of wax matrix and microencapsulated potassium chloride in relation to upper gastrointestinal illness requiring hospitalization.
The frequency of hospitalization for documented upper gastrointestinal illness among recipients of either wax matrix or microencapsulated potassium chloride was similar in a study in which clinics were randomly assigned to dispense one or the other preparation. The results indicate that there is no material difference between the two preparations in relation to serious upper gastrointestinal illness.


Growth of Trypanosoma cruzi epimastigotes controlled by shifts in cyclic AMP mediated by adrenergic ligands.
Intracellular levels of cAMP in Trypanosoma cruzi epimastigotes were determined in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, and in the presence of the adrenergic ligand isoproterenol. An increase in the levels of cAMP was observed when those drugs were added. This effect was more pronounced when the cells were in the lag phase of growth. Phosphodiesterase inhibitors, catecholamines and dibutyryl cAMP inhibited proliferation of the organisms, as well as [3H]thymidine incorporation in in vitro experiments. These findings strongly suggest that cAMP plays a role in the control of growth of T. cruzi. The effects of adrenergic ligands in increasing cAMP levels and inhibiting growth could be reversed by beta-adrenergic antagonists, suggesting the presence of a receptor-mediated adenylate cyclase system in T. cruzi. Cholinergic ligands had no effect.


Trait or testimony? Children's preferences for positive informants.
Research indicates that children often show a positivity bias, or a tendency to favor positive information over negative information, in assessments of informant credibility in social and nonsocial situations. The current study investigated whether young children prioritize positive informant traits (i.e., nice vs. mean informant) as compared with positive speech content (i.e., positive vs. negative evaluation) in conflicting assessments of a work product. A total of 123 4- to 8-year-olds heard stories about a nice informant who gave a negative evaluation of a painting and a mean informant who gave a positive evaluation of the painting. Participants were asked who they would endorse, who they would ask about a future painting, and their friendship preferences. Children endorsed and asked the mean informant who provided positive testimony, but they chose to befriend the nice informant who provided negative testimony. Endorsements of positive testimony increased with age. Findings are considered in the context of the broader literature on selective social learning and trait understanding.


Sensitive and specific method for the determination of josamycin in human plasma by liquid chromatography-mass spectrometry.
A highly sensitive and specific method for the determination of josamycin in human plasma by LC-MS was developed and validated. Josamycin was extracted from human plasma by a single-step liquid-liquid extraction and analyzed by LC-MS via an electrospray ionization interface. Selected ion monitoring was used to detect josamycin and its internal standard. The intra-day precision and accuracy, expressed as C.V. and R.E., ranged from 2.8% to 13.5% and -10.3% to 7.6%, respectively. The lower limit of detection was 0.1 ng/ml and the lower limit of quantitation was set at 1 ng/ml when 0.5 ml of plasma was used. No endogenous interference was observed in human plasma obtained from drug-free volunteers.


Prevalence of and Risk Factors for Multidrug-Resistant Bacteria in Urine Cultures of Spinal Cord Injury Patients.
To identify the prevalence of multidrug-resistant (MDR) bacteria and identify their specific risk factors in routine urine specimens of spinal cord injury (SCI) patients. This study was designed in a retrospective manner, reviewing the medical records of SCI patients who were admitted to a specialized SCI unit between January 2001 and December 2013. Patients were investigated for age, gender, American Spinal Injury Association impairment scale, SCI level, duration after injury, bladder management method, and hospitalization history within four weeks prior to visiting our unit. The results of routine urine cultures including presence of MDR organisms were analyzed. Among the total 2,629 urine samples from the newly admitted SCI patients, significant bacteriuria was identified in 1,929 (73.4%), and MDR organisms were isolated in 29 (1.1%) cultures. There was an increasing trend of MDR organism prevalence from 2001 to 2013 (p<0.01). The isolation of MDR organisms in inpatients who were admitted for rehabilitation (1.3%) was significantly higher than it was among community-residing persons (0.2%) (p<0.05). By voiding method, patients who used a suprapubic indwelling catheter (3.3%) or a urethral indwelling catheter (2.6%) showed a higher rate of MDR organism isolation (p<0.05). There was an increasing trend of MDR organism isolation in SCI patients. Inpatients and persons who used indwelling catheters showed a higher risk of MDR organism isolation.


Poly(aspartic acid)-based self-healing hydrogel with precise antibacterial ability for rapid infected-wound repairing.
The development of wound dressings with antibacterial activities and simultaneous pro-healing functions are always urgent in treating bacterial wound infection. Herein, a novel multifunctional self-healing hydrogel was designed and prepared by crosslinking quaternary ammonium/boronic acid modified poly(aspartic acid) and poly (vinyl alcohol) polymers with targeted peptide MP196- conjugated polydopamine. The formation of this hydrogel not only improves the biocompatibility of quaternary poly(aspartic acid), but also enhances antibacterial efficacy by pH-triggering dissociation under the low pH bacterial microenvironment. Moreover, precise photothermal treatment can be achieved. In vitro study suggested high synergistic antibacterial efficiency(∼100 %) under near-infrared light, significantly higher than a single antibacterial strategy (66.0-82.6 %). In vivo study suggested infected wounds treated with the hydrogel showed an optimal healing rate(92.0 %) after 7 days. The survival rate of the bacteria in the epidermal tissues was reduced to 2.3 %. Besides, the suitable self-healing property of this hydrogel facilitated its application in the diversity of wound shapes. Thus, the novel poly(aspartic acid) hydrogel might be a promising candidate for precise therapy of bacteria-infected wounds.


Development of new methods to determine work disability in the United States.
The development of new methods to determine work disability for the United States Social Security Administration is described, including the fiscal and administrative background to the current and proposed methods. An introduction to the current disability determination process and description of its status is followed by a description of the original proposed plan for redesign of the process. In response to this plan, the authors participated in several research projects. An overview of some of the key research projects performed to improve the Social Security Administration disability determination process is provided.


Phlaeopterus Motschulsky, 1853 (Coleoptera: Staphylinidae: Omaliinae: Anthophagini)--a new genus for the Palaearctic: new combination.
The genus Phlaeopterus Motschulsky, 1853, which was previously known only from North America, is recorded for the Palaearctic fauna for the first time: P. czerskyi (Shavrin, 2001) comb. nov. (East Siberia: Khamar-Daban Mts.) is transferred from the genus Lesteva Latreille, 1797, redescribed, and illustrated.


Charge transfer and first hyperpolarizability: cage-like radicals C59X and lithium encapsulated Li@C59X (X=B, N).
Very recently, two new cage-like radicals (C59B and C59N) formed by a boron or nitrogen atom substituting one carbon atom of C60 were synthesized and characterized. In order to explore the structure-property relationships of combination the cage-like radical and alkali metal, the endohedral Li@C59B and Li@C59N are designed by lithium (Li) atom encapsulated into the cage-like radicals C59B and C59N. Further, the structures, natural bond orbital (NBO) charges, and nonlinear optical (NLO) responses of C59B, C59N, Li@C59B, and Li@C59N were investigated by quantum chemical method. Three density functional methods (BHandHLYP, CAM-B3LYP, and M05-2X) were employed to estimate their first hyperpolarizabilities (β tot) and obtained the same trend in the β tot value. The β tot values by BHandHLYP functional of the pure cage-like radicals C59B (1.30 × 10(3) au) and C59N (1.70 × 10(3) au) are close to each other. Interestingly, when one Li atom encapsulated into the electron-rich radical C59N, the β tot value of the Li@C59N increases to 2.46 × 10(3) au. However, when one Li atom encapsulated into the electron-deficient radical C59B, the β tot value of the Li@C59B sharply decreases to 1.54 × 10(2) au. The natural bond orbital analysis indicates that the encapsulated Li atom leads to an obvious charge transfer and valence electrons distribution plays a significant role in the β tot value. Further, frontier molecular orbital explains that the interesting charge transfer between the encapsulated Li atom and cage-like radicals (C59B and C59N) leads to differences in the β tot value. It is our expectation that this work will provide useful information for the design of high-performance NLO materials.


Human CHAC1 Protein Degrades Glutathione, and mRNA Induction Is Regulated by the Transcription Factors ATF4 and ATF3 and a Bipartite ATF/CRE Regulatory Element.
Using an unbiased systems genetics approach, we previously predicted a role for CHAC1 in the endoplasmic reticulum stress pathway, linked functionally to activating transcription factor 4 (ATF4) following treatment with oxidized phospholipids, a model for atherosclerosis. Mouse and yeast CHAC1 homologs have been shown to degrade glutathione in yeast and a cell-free system. In this report, we further defined the ATF4-CHAC1 interaction by cloning the human CHAC1 promoter upstream of a luciferase reporter system for in vitro assays in HEK293 and U2OS cells. Mutation and deletion analyses defined two major cis DNA elements necessary and sufficient for CHAC1 promoter-driven luciferase transcription under conditions of ER stress or ATF4 coexpression: the -267 ATF/cAMP response element (CRE) site and a novel -248 ATF/CRE modifier (ACM) element. We also examined the ability of the CHAC1 ATF/CRE and ACM sequences to bind ATF4 and ATF3 using immunoblot-EMSA and confirmed ATF4, ATF3, and CCAAT/enhancer-binding protein β binding at the human CHAC1 promoter in the proximity of the ATF/CRE and ACM using ChIP. To further validate the function of CHAC1 in a human cell model, we measured glutathione levels in HEK293 cells with enhanced CHAC1 expression. Overexpression of CHAC1 led to a robust depletion of glutathione, which was alleviated in a CHAC1 catalytic mutant. These results suggest an important role for CHAC1 in oxidative stress and apoptosis with implications for human health and disease.


Laser capture microdissection coupled mass spectrometry (LCM-MS) for spatially resolved analysis of formalin-fixed and stained human lung tissues.
Haematoxylin and eosin (H&E)-which respectively stain nuclei blue and other cellular and stromal material pink-are routinely used for clinical diagnosis based on the identification of morphological features. A richer characterization can be achieved by laser capture microdissection coupled to mass spectrometry (LCM-MS), giving an unbiased assay of the proteins that make up the tissue. However, the process of fixing and H&E staining of tissues provides challenges with standard sample preparation methods for mass spectrometry, resulting in low protein yield. Here we describe a microproteomics technique to analyse H&E-stained, formalin-fixed paraffin-embedded (FFPE) tissues. Herein, we utilize heat extraction, physical disruption, and in column digestion for the analysis of H&E stained FFPE tissues. Micro-dissected morphologically normal human lung alveoli (0.082 mm3) and human lung blood vessels (0.094 mm3) from FFPE-fixed H&E-stained sections from Idiopathic Pulmonary Fibrosis (IPF) specimens (n = 3 IPF specimens) were then subject to a qualitative and then quantitative proteomics approach using BayesENproteomics. In addition, we tested the sensitivity of this method by processing and analysing a range of micro-dissected human lung blood vessel tissue volumes. This approach yields 1252 uniquely expressed proteins (at a protein identification threshold of 3 unique peptides) with 892 differentially expressed proteins between these regions. In accord with prior knowledge, our methodology approach confirms that human lung blood vessels are enriched with smoothelin, CNN1, ITGA7, MYH11, TAGLN, and PTGIS; whereas morphologically normal human lung alveoli are enriched with cytokeratin-7, -8, -18, -19, 14, and -17. In addition, we identify a total of 137 extracellular matrix (ECM) proteins and immunohistologically validate that laminin subunit beta-1 localizes to morphologically normal human lung alveoli and tenascin localizes to human lung blood vessels. Lastly, we show that this micro-proteomics technique can be applied to tissue volumes as low as 0.0125 mm3. Herein we show that our multistep sample preparation methodology of LCM-MS can identify distinct, characteristic proteomic compositions of anatomical features within complex fixed and stained tissues.


Factors influencing lipoprotein lipase activity in choline deficiency.
Subcutaneous injections of the lipotropic agent, ethyl trichloracetate, to rats with established choline deficiency raised their plasma triglycerides by 60% and completely removed the hyperglyceridaemic response of Triton WR 1339. The plasma triglyceride levels of choline-supplemented rats were depressed slightly by ethyl trichloracetate administration, which was effective in abolishing response to Triton WR 1339. Lipoprotein lipase activity of epididymal fat pad was stimulated 60% while plasma lipoprotein was not stimulated by ethyl trichloracetate. The increased peripheral removal of low-density lipoprotein-triglyceride complex, allowing greater use to be made of existing apo-proteins, may explain the lipotropic character of the ester.


The use of statistical methods for censored data to evaluate the activity concentration of Pb-210 in beans (Phaseolus vulgaris L.).
A survey of 210Pb activity concentration, one of the major internal natural radiation sources to man, has been carried in the most common species of beans (Phaseolus vulgaris L.) grown and consumed in Brazil. The representative bean types chosen, Carioca beans and black type sown in the Brazilian Midwestern and Southern regions, have been collected in this study and 210Pb determined by liquid scintillation spectrometry after separation with chromatographic extraction using Sr-resin. Available values in data set of radioactivity in Brazil (GEORAD) on the 210Pb activity concentration in black beans grown in Southeastern region have been added to the results of this study with the purpose of to amplify the population considered. Concerning the multiple detection limits and due to the high level of censored observations, a robust semi-parametric statistical method called regression on order statistics (ROS) has been employed to provide a reference value of the 210Pb in Brazilian beans, which amounted to 41 mBq kg-1 fresh wt. The results suggest that the 210Pb activity concentration in carioca beans is lower than in black beans. Also evaluated was the 210Pb activity concentration in vegetable component of a typical diet, which displays lower values than those shown in the literature for food consumed in Europe.


Loss of CRABP-II Characterizes Human Skin Poorly Differentiated Squamous Cell Carcinomas and Favors DMBA/TPA-Induced Carcinogenesis.
Retinol and its derivatives play an important role in epidermal growth and differentiation and represent chemopreventive agents in nonmelanoma skin cancer. Retinoic acid binding protein II (CRABP-II) is a cytoplasmic receptor that critically regulates all-trans-retinoic acid (ATRA) trafficking. We documented the marked reduced expression of CRABP-II and its promoter methylation in human poorly differentiated squamous cell carcinomas. To investigate the role of CRABP-II in skin carcinogenesis we used skin lesion induction by dimethylbenz[a]anthracene/12-O-tetradecanoyl-phorbol-13-acetate in CRABP-II-knockout C57BL/6 mice. We observed earlier and more diffuse epidermal dysplasia, greater incidence and severity of tumors, reduced expression of cytokeratin 1/cytokeratin 10 and involucrin, increased proliferation, and impaired ATRA inhibition of tumor promotion compared with wild-type animals. CRABP-II-transfected HaCaT, FaDu, and A431 cells showed expression of differentiation markers, retinoic acid receptor-β/-γ signaling, ATRA sensitivity, and suppression of EGFR/v-akt murine thymoma viral oncogene homolog 1 (AKT) pathways in a fatty acid binding protein 5/peroxisome proliferator-activated receptor-β/-δ-independent manner. The opposite was true in keratinocytes isolated from CRABP-II-knockout mice. Finally, CRABP-II accumulation induced ubiquitination-associated reduction of EGFR. Our results showed reduced CRABP-II expression in human poorly differentiated squamous cell carcinomas, and its gene deletion favored experimental skin carcinogenesis and impaired ATRA antitumor efficacy, likely modulating EGFR/AKT pathways and retinoic acid receptor-β/-γ signaling. Therapeutic interventions aimed at restoring CRABP-II-mediated signaling may amplify therapeutic retinoid efficacy in nonmelanoma skin cancer.


Correction: Preparation of a novel injectable in situ-gelling nanoparticle with applications in controlled protein release and cancer cell entrapment.
[This corrects the article DOI: 10.1039/C8RA06589F.].


tert-Butyl 3-amino-5-bromo-1H-indazole-1-carboxyl-ate.
In the title compound, C12H14BrN3O2, the pyrazole and benzene rings are nearly co-planar with a dihedral angle between the rings of 2.36 (5)°. In the crystal, inversion dimers linked by pairwise N-H⋯N hydrogen bonds generate R 2 2(8) loops. The dimers are linked into a three-dimensional network by weak aromatic π-π stacking inter-actions [centroid-centroid separation = 3.7394 (6) Å] and C-H⋯O and C-H⋯Br hydrogen bonds.


[Perplexing acute or chronic somatic or psychiatric symptoms: possibly due to porphyria].
A woman aged 35 years had a classical picture of acute intermittent porphyria. However, the diagnosis was only made after repeated clinical investigations by internists, gastroenterologists, gynaecologists and surgeons working at a university hospital, and after an unnecessary laparotomy. Recent data indicate that acute intermittent porphyria is not a very rare disease. It may run a serious invalidating and life-threatening course. Its manifestations vary and may lead to very different clinical pictures.


An alternative method for reconstruction of large intranasal lining defects: the Farina method revisited.
To examine use of the paramedian forehead flap for intranasal lining. The medical records and photographs were reviewed for patients who underwent a nasal reconstruction involving a paramedian forehead flap used for internal lining. Three patients underwent this procedure. In all patients, the flap provided excellent intranasal lining. External incisions were acceptable in all the patients. The paramedian forehead flap may be used for internal lining of large intranasal defects. An external rhinoplasty approach facilitates reconstruction.


Ultrastructural aspects of platelet adhesion on subendothelial structures.
Platelet adhesion to ligands within the subendothelium induces platelet activation. Ultrastructural and immunocytochemical investigations were carried out to obtain information on the microtopography of the adhesive interactions. Rabbit platelets which react in vivo with endothelial lesions (V. jugularis), and human platelets which react in vitro with collagen were observed. It was shown that the reaction of adhering platelets depends on the micro-topography of the adhesive surface 1. On plane surfaces the platelets spread 2. Collagen fibers induce the formation of focal contacts. They are initiated by binding of ligands to certain transmembrane receptors. The contact mediates the formation of the contractile gel, which acts as a constricting sphere internalizing surface bound ligands, i.e., retraction of collagen networks. Most of the experiments hitherto dealt with platelets that spread. The described findings show that the reaction mentioned above plays a more important role than previously imagined and may be the physiological response of platelets interacting with subendothelial components in vessel lesions.


Anticonvulsant activity of GABA uptake inhibitors and their prodrugs following central or systemic administration.
The activity of compounds inhibiting neuronal or glial GABA uptake has been assessed following intracerebroventricular (i.c.v.) or intraperitoneal (i.p.) administration in DBA/2 mice (sound-induced seizures) or Swiss S mice (pentylenetetrazol-induced seizures). Sound-induced seizures are suppressed by the i.c.v. injection of (+/-)-nipecotic acid, 3.2 mumol, or (+/-)-cis-4-hydroxynipecotic acid, 2 mumol, but not by i.p. injection of (+/-)-nipecotic acid, 3.2 mmol/kg or (+/-)-cis-4-hydroxynipecotic acid 4 mmol/kg. Pentylenetetrazol-induced seizures are not suppressed by i.c.v. injection of (+/-)-nipecotic acid 1-4 mumol, or (+/-)-cis-4-hydroxynipecotic acid, 2-4 mumol. THPO (4,5,6,7-tetrahydroisoxazolo[4.5-c]pyridin-3-ol), 1-5 mumol i.c.v. or 1-4 mmol/kg i.p., protects against sound-induced seizures. There is no protection against pentylenetetrazol seizures after i.c.v. THPO injection, but THPO, 2-8 mmol/kg i.p., is protective. Among prodrugs, (+/-)-nipecotic acid pivaloyloxymethyl ester protects against sound-induced seizures, when given i.c.v. (3.2 mumol) or i.p. (1.6-3.2 mmol/kg) and against pentylenetetrazol seizures when given i.p. (0.5-4 mmol/kg). (+/-)-cis-4-hydroxynipecotic acid methyl ester protects against sound-induced seizures when given i.p. (3.2 mmol/kg), but is only partially protective against pentylenetetrazol seizures, when given i.p. (4 mmol/kg). Some prodrugs induce myoclonus following either i.c.v. or i.p. administration.