Characteristics and outcomes of lung transplants performed with ex-situ lung perfusion.
Ex-situ lung perfusion (ESLP) can be used to assess and rehabilitate donor lungs, potentially expanding the donor pool. We examined the characteristics and outcomes of lung transplants performed with ESLP in the United States. Retrospective review of the United Network for Organ Sharing registry of primary adult lung transplant recipients from February 28, 2018, to June 30, 2021, was performed, comparing baseline characteristics, in-hospital outcomes, and 1-year survival of ESLP vs no ESLP lung transplants. Of 8204 lung transplants, 426 (5.2%) were performed with ESLP. ESLP donors were older, more donation after circulatory death (DCD), and had lower PaO2:FiO2 (P:F) ratios. Recipients had lower lung allocation scores. ESLP lungs traveled further, had longer preservation times, and were more likely double lung transplants. Reintubation rates, extracorporeal membrane oxygenation at 72 hours, and hospital length of stay were greater in the ESLP group. On multivariable analysis, ESLP was not an independent predictor of 1-year survival. However, further analysis showed that DCD lungs managed on ESLP had worse 1-year survival compared to DCD lungs preserved with standard cold storage or with donation after brain death donor lungs. ESLP is used in a small percentage of lung transplants in the US and is not independently associated with 1-year survival. ESLP combined with DCD lungs, however, is associated with worse 1-year survival and warrants further investigation.


AICAR induces Bax/Bak-dependent apoptosis through upregulation of the BH3-only proteins Bim and Noxa in mouse embryonic fibroblasts.
5-Aminoimidazole-4-carboxamide (AICA) riboside (AICAR) is a nucleoside analogue that is phosphorylated to 5-amino-4-imidazolecarboxamide ribotide (ZMP), which acts as an AMP mimetic and activates AMP-activated protein kinase (AMPK). It has been recently described that AICAR triggers apoptosis in chronic lymphocytic leukemia (CLL) cells, and its mechanism of action is independent of AMPK as well as p53. AICAR-mediated upregulation of the BH3-only proteins BIM and NOXA correlates with apoptosis induction in CLL cells. Here we propose mouse embryonic fibroblasts (MEFs) as a useful model to analyze the mechanism of AICAR-induced apoptosis. ZMP formation was required for AICAR-induced apoptosis, though direct Ampk activation with A-769662 failed to induce apoptosis in MEFs. AICAR potently induced apoptosis in Ampkα1 (-/-) /α2 (-/-) MEFs, demonstrating an Ampk-independent mechanism of cell death activation. In addition, AICAR acts independently of p53, as MEFs lacking p53 also underwent apoptosis normally. Notably, MEFs lacking Bax and Bak were completely resistant to AICAR-induced apoptosis, confirming the involvement of the mitochondrial pathway in its mechanism of action. Apoptosis was preceded by ZMP-dependent but Ampk-independent modulation of the mRNA levels of different Bcl-2 family members, including Noxa, Bim and Bcl-2. Bim protein levels were accumulated upon AICAR treatment of MEFs, suggesting its role in the apoptotic process. Strikingly, MEFs lacking both Bim and Noxa displayed high resistance to AICAR. These findings support the notion that MEFs are a useful system to further dissect the mechanism of AICAR-induced apoptosis.


Provenances, preponderances, and distribution of humic acids and organic pollutants in hydro-geosphere: The co-existence, interaction and isotopic biomarkers in the riverine ecosystem.
This paper aims to critically review the importance of geochemical fingerprinting and tracing using biomarkers and stable isotopes in the riverine ecosystem and depicts that isotopic ratios of δ13C, δ15N, and δ34S can be used for tracing pollution sources. Stable isotopes like carbon, hydrogen, nitrogen, oxygen, and sulfur are being used for this purpose, and their isotopic signatures are primarily used to distinguish close sources of organic matter through dual isotopes. The present review is articulated to bridge the critical research gaps of the previous and contemporary documented literature on the genesis and transport of OM between freshwater and marine systems. This review comprehensively provides methods and techniques in geochemical tracing and discusses the future directions to address the challenges of the current methods to enhance the knowledge about the source identification of organic matter in the riverine environment. Tracer geochemistry emphasizes the implications of elemental abundances and isotope ratio variations in geologic substances to track natural earth processes, anthropogenic contaminants, and geochemical signatures in the hydrologic system. The principal constituent of organic matter comprises humic substances like humic acid, fulvic acid, and humin, and these comprise 50-75% of the sediments and DOC in natural waters. Their structural and functional characterization is required to elucidate the transport and fate of organic matter, which are often influenced by several paleoenvironmental factors.


Transformation of the 8-9 S molybdate-stabilized estrogen receptor from low-affinity to high-affinity state without dissociation into subunits.
The rate of dissociation of labeled estradiol from [3H] estradiol-8-9 S receptor complexes ([3H]E2-8-9 S ER) molybdate-stabilized was determined in the presence of either an excess of unlabeled hormone ("chase") or of charcoal/dextran suspension ("stripping"). Biphasic dissociation of the hormone was observed in both cases, but the fraction of the fast-dissociating component was dramatically reduced (5% instead of 60%) when stripping was used. As the dissociation patterns were independent of the degree of saturation of the receptor, the results do not favor the possibility of cooperative effects between binding sites in the 8-9 S ER. After pretreatment of cytosol by charcoal at 28 degrees C for 15 min, the dissociation studied by chase displayed only the slowly dissociating component (t1/2 approximately 65 min). This effect was dependent on temperature and influenced by the ligand bound to 8-9 S ER, being pronounced with estradiol (E2) and absent with [3H]4-hydroxytamoxifen. The slow-dissociating component obtained after charcoal treatment was reconverted to fast-dissociating state by adding dithiothreitol or by incubation with cytosol at 20 degrees C. The charcoal treatment did not change the sedimentation coefficient (approximately 9 S) and the Stokes radius (approximately 7 nm) of the [3H]E2-8-9 S ER, and the slow-dissociating form obtained did not bind to DNA-cellulose either in the presence or absence of molybdate ions. Thus there are likely small but functionally significant changes of structure in the 8-9 S ER which remain in a non-DNA-binding form, whereas the rate of estradiol dissociation is modified.


Oxidative imbalance in idiopathic renal hypouricemia.
An important complication of idiopathic renal hypouricemia is exercise-induced acute renal failure (ARF). The most plausible explanation for this complication is that decreased antioxidant potential leads to kidney injury by reactive oxygen species (ROS). We demonstrated this oxidative imbalance by a concomitant assessment of ROS production and antioxidant system capability in a 15- year-old girl with idiopathic renal hypouricemia caused by a mutation in the urate transporter (URAT1) gene. Her serum level of ROS increased with decreasing antioxidant potential capacity soon after the initiation of anaerobic stress due to treadmill exercise. Thereafter, serum levels of ROS and antioxidant potential showed a parallel course, returning to the baseline values at 240 min after exercise. Some patients with idiopathic renal hypouricemia demonstrate oxidative imbalance soon after exercise with a predisposition to exercise-induced acute renal failure. Antioxidant properties may alter this imbalance by augmenting the antioxidant activity.


Correction to "Maternal vitamin D status in relation to infant BMI growth trajectories up to 2 years of age in two prospective pregnancy cohorts".
[This corrects the article DOI: 10.1002/osp4.602.].


Neurodevelopmental Outcome of Extremely Low Birth Weight Children at Corrected Age of Two Years.
To assess the neurodevelopmental, cognitive and behavioral function of extremely low birth weight babies (ELBW) till corrected age of two years. 79 ELBW babies were enrolled and followed at 1 year (n=50), 18 months (n=47) and 2 years (n=36). Adverse composite outcome was defined as death or moderate-to-severe neurodevelopmental impairment (defined as either cerebral palsy or DQ score <70 or deafness or blindness). At 1 year, 24% were neurologically abnormal. At 18 months, average score (>85) was seen in 25 (54%) children in motor and 8 (17%) in mental development. Abnormal behavioral score (?12) was seen in 89% children. Adverse composite outcome was present in 28 (35.4%) babies. ELBW neonates are at a high risk of neurodevelopmental and behavioral impairment.


Perceived timing of first- and second-order changes in vision and hearing.
Simultaneous changes in visual stimulus attributes (such as motion or color) are often perceived to occur at different times, a fact usually attributed to differences in neural processing times of those attributes. However, other studies suggest that perceptual misalignments are not due to stimulus attributes, but to the type of change, first- or second-order. To test whether this idea generalizes across modalities, we studied perceptual synchrony of acoustic and of audiovisual cross-modal stimuli, which varied in a first- or second-order fashion. First-order changes were abrupt changes in tone intensity or frequency (auditory), or spatial position (visual), while second-order changes were an inversion of the direction of change, such as a turning point when a rising tone starts falling or a translating visual blob reverses. For both pure acoustic and cross-modal stimuli, first-order changes were systematically perceived before second-order changes. However, when both changes were first-order, or both were second-order, little or no difference in perceptual delay was found between them, regardless of attribute or modality. This shows that the type of attribute change, as well as latency differences, is a strong determinant of subjective temporal alignments. We also performed an analysis of reaction times (RTs) to the first- and second-order attribute changes used in these temporal alignment experiments. RT differences between these stimuli did not correspond with our temporal alignment data, suggesting that subjective alignments cannot be accounted for by a simple latency-based explanation.


Stereotactic amygdalohippocampectomy for temporal lobe epilepsy: promising results in 16 patients.
Minimally invasive procedures for treating temporal lobe epilepsy have been investigated recently, namely stereotactic and gamma knife amygdalohippocampectomy (AHE). However, the results are not fully satisfactory. Our aim was to evaluate efficacy and side-effects of stereotactic AHE mimicking the neurosurgical procedure in terms of extent of the lesion. 16 consecutive patients were assessed using VEEG, MRI, FDG-PET and WADA test. All had definite pharmacoresistant medial temporal lobe epilepsy. The stereotactic AHE was performed on the Leksell stereotactic system. All lesions exceeded 40 mm along the long axis of the hippocamus. Seizure outcome was favourable on one year follow-up: 12 patients (75%) were seizure-free (Engel I), three (19%) were Engel II, and one (6%) was Engel III. Side-effects were mild, lasting up to 7 days: cephalea, meningeal syndrome with sterile CSF in three subjects, and CSF leak lasting up to 3 days in seven subjects. Stereotactic AHE encompassing sufficient volume of the amygdalohippocampal complex appears to be safe, effective, and free from long-term side-effects.


Exercise oximetry versus spirometry in the assessment of risk prior to lung resection.
Spirometry remains a standard method of assessing patient risk prior to lung resection despite its poor sensitivity and specificity. This study compares the relative ability of standardized exercise oximetry and spirometry--forced expiratory volume in the first second--to predict morbidity and mortality after lung resection. The study comprised a retrospective review of 396 consecutive patients of whom 299 underwent both oximetry and spirometry. Oximetry was undertaken during standard exercise under the supervision of a single physical therapist. Spirometry identified 46 patients with a forced expiratory volume in the first second of less than 1.5 L who were considered to be high risk. Exercise oximetry was used to identify patients with arterial oxygen desaturation at rest, while walking on level ground, or while climbing two flights of stairs (n = 65). Compared with spirometry, exercise oximetry more reliably predicted home oxygen requirements (p < 0.001), need of admission to the intensive care unit (p < 0.05), prolonged hospital stay (p < 0.001), and respiratory failure (p < 0.05). Oximetry identified 50% of the patients who died, all of whom had a forced expiratory volume in the first second of greater than 1.5 L. Despite its superior predictive value, the sensitivity of oximetry remained low. We conclude that standardized exercise oximetry is a superior screen of the high-risk patient than spirometry (forced expiratory volume in the first second) prior to pulmonary resection when there are no other risk factors noted on initial history and physical examination. A prospective, randomized trial is required to substantiate this conclusion.


Acetylation of the HIV-1 Tat protein: an in vitro study.
In the last few years, the understanding of lysine acetylation as a regulatory post-translational modification of proteins in cell signalling cascades has increased. It is now known that not only histones but also non-histone factors can serve as substrates of different acetyltransferase enzymes. Acetylated lysine residues in non-histone factors are often identified using radioactive labelling experiments and immunochemical analysis of synthetic peptides. In this study of the human immunodeficiency virus 1 (HIV-1) Tat protein, we demonstrate the benefits of matrix-assisted laser desorption/ionisation mass spectrometry, proteolytic digestion and Edman sequencing for the mapping of acetylation sites. We confirmed that the HIV-1 Tat protein is acetylated in vitro by the acetyltransferase p300 at a specific lysine residue at position 50 in its RNA binding region. Furthermore, we showed that the Tat cysteine-rich region is acetylated at multiple cysteine residues in the absence of enzyme. Since this non-enzymatic cysteine acetylation occurs independently from the surrounding peptide sequence, we consider the presence of cysteine residues in acetylated peptides an important factor for the interpretation of in vitro acetylation assays in general.


[Investigation of the effect of various buffer solutions used for microinjection of gene-engineering constructs on preimplantation development of mouse embryos].
We studied the effects of some buffer solutions used for microinjection in mammalian zygotes on preimplantation development of (CBA x C57BL)F1 mouse embryos in vitro. The rate of embryo survival was estimated according to their capacity to develop to the stages of blastocyst and blastocyst hatched from zona pellucida. The results obtained suggested a reduced rate of survival of zygotes to the blastocyst stage after the injection into a pronucleus of the buffers Tris-HCl with EDTA, Tris-HCl with MgCl2 and NaCl, and medium M2 (p < 0.05) and to the stage of blastocyst hatched from zone pellucida after injection of a Dulbecco solution, as compared to the control. No differences were found in the survival rate of zygotes injected with different buffer solutions.


Tolerance and cross-tolerance to the rate-suppressing effects of opioids in butorphanol-treated rats: influence of maintenance dose and relative efficacy at the mu receptor.
The purpose of the present investigation was to examine the development of tolerance to the rate-suppressing effects of mu and kappa opioids in rats administered either 3.0 (low) or 30 (high) mg/kg per day of butorphanol, an opioid with low relative efficacy at the mu receptor. The mu opioids butorphanol, buprenorphine, morphine, fentanyl and sufentanil, and the kappa opioid U50,488 dose-dependently suppressed responding under all conditions examined. In rats administered the low maintenance dose of butorphanol, tolerance developed to the effects of butorphanol, buprenorphine and morphine, but not to fentanyl and sufentanil. In rats administered the high maintenance dose, tolerance developed to all of the mu opioids examined. In both treatment groups, the degree to which tolerance developed was greater for butorphanol and buprenorphine than for morphine, fentanyl and sufentanil; and the degree to which tolerance developed to these mu opioids was greater in rats administered the high maintenance dose of butorphanol. The tolerance that developed to morphine, fentanyl and sufentanil was not altered when tested at both 23 and 47 h following the previous maintenance dose of butorphanol, suggesting that these changes were not due to any acute pharmacological interactions between butorphanol and the test compound (i.e., antagonism). Tolerance was also conferred to the kappa opioid U50,488 in both groups of rats, and in rats administered the high maintenance dose, this effect was obtained when tested 23 and 47 h following the previous maintenance dose of butorphanol. Physical dependence developed in rats administered the high maintenance dose of butorphanol, as evidenced by the development of enhanced sensitivity to the rate-suppressing effects of naloxone, and the finding that 30 mg/kg naloxone decreased body weight in a time-dependent manner. No physical dependence was apparent in rats administered the low maintenance dose of butorphanol. These data suggest that during chronic treatment with butorphanol, (1) greater degrees of tolerance are conferred to drugs possessing low efficacy at the mu opioid receptor, (2) tolerance is enhanced as the maintenance dose of the toleragen is increased, and (3) mu-opioid tolerance may be observed under conditions that do not produce mu-opioid dependence.


nMAGMA: a network-enhanced method for inferring risk genes from GWAS summary statistics and its application to schizophrenia.
Annotating genetic variants from summary statistics of genome-wide association studies (GWAS) is crucial for predicting risk genes of various disorders. The multimarker analysis of genomic annotation (MAGMA) is one of the most popular tools for this purpose, where MAGMA aggregates signals of single nucleotide polymorphisms (SNPs) to their nearby genes. In biology, SNPs may also affect genes that are far away in the genome, thus missed by MAGMA. Although different upgrades of MAGMA have been proposed to extend gene-wise variant annotations with more information (e.g. Hi-C or eQTL), the regulatory relationships among genes and the tissue specificity of signals have not been taken into account. We propose a new approach, namely network-enhanced MAGMA (nMAGMA), for gene-wise annotation of variants from GWAS summary statistics. Compared with MAGMA and H-MAGMA, nMAGMA significantly extends the lists of genes that can be annotated to SNPs by integrating local signals, long-range regulation signals (i.e. interactions between distal DNA elements), and tissue-specific gene networks. When applied to schizophrenia (SCZ), nMAGMA is able to detect more risk genes (217% more than MAGMA and 57% more than H-MAGMA) that are involved in SCZ compared with MAGMA and H-MAGMA, and more of nMAGMA results can be validated with known SCZ risk genes. Some disease-related functions (e.g. the ATPase pathway in Cortex) are also uncovered in nMAGMA but not in MAGMA or H-MAGMA. Moreover, nMAGMA provides tissue-specific risk signals, which are useful for understanding disorders with multitissue origins.


Safety of Propofol versus Nonpropofol-Based Sedation in Children Undergoing Gastrointestinal Endoscopy: A Systematic Review and Meta-Analysis.
The majority of children who undergo gastrointestinal (GI) endoscopy require anesthesia or procedural sedation for comfort, cooperation, and procedure efficiency. The safety profile of propofol is not well established in children but has been studied in the literature. The aim of this study is to evaluate and compare the safety of propofol-only sedation for GI endoscopy procedures to other anesthetic regimes in the pediatric population. A search was conducted in the MEDLINE, Embase, and Cochrane Library databases. Randomized clinical trials and prospective cohorts were included in the study. No significant difference was noted in total complications between the two cohorts with a pooled OR of 1.31 (95% CI: 0.57-3.04, chi2 = 0.053, I2 = 54.31%). The pooled rate of complications in the studies was 23.4% for those receiving propofol only and 18.2% for those receiving other anesthetic regimens. Sensitivity analysis was performed removing a study with a very different control comparison compared to the rest of the studies included. Once excluded, there was minimal heterogeneity in the remaining studies and a significant difference in overall complications was detected, with more complications seen in the propofol-only group compared to the other anesthetic groups (OR 1.87, 95% CI 1.09-3.20). Significantly higher incidence of cardiorespiratory complications was noted in the propofol-only versus other anesthetic regimens in pediatric patients undergoing GI endoscopy in this meta-analysis. However, the overall quality of the evidence is very low. Clinicians providing sedation to a pediatric population for GI endoscopy should consider there may be increased risks when using a propofol-only regimen, but further study is needed.


Primary payer status is significantly associated with postoperative mortality, morbidity, and hospital resource utilization in pediatric surgical patients within the United States.
Current healthcare reform efforts have highlighted the potential impact of insurance status on patient outcomes. The influence of primary payer status (PPS) within the pediatric surgical patient population remains unknown. The purpose of this study was to examine risk-adjusted associations between PPS and postoperative mortality, morbidity, and resource utilization in pediatric surgical patients within the United States. A weighted total of 153,333 pediatric surgical patients were evaluated using the national Kids' Inpatient Database (2003 and 2006): appendectomy, intussusception, decortication, pyloromyotomy, congenital diaphragmatic hernia repair, and colonic resection for Hirschsprung's disease. Patients were stratified according to PPS: Medicare (n=180), Medicaid (n=51,862), uninsured (n=12,539), and private insurance (n=88,753). Multivariable hierarchical regression modeling was utilized to evaluate risk-adjusted associations between PPS and outcomes. Overall median patient age was 12 years, operations were primarily non-elective (92.4%), and appendectomies accounted for the highest proportion of cases (81.3%). After adjustment for patient, hospital, and operation-related factors, PPS was independently associated with in-hospital death (p<0.0001) and postoperative complications (p<0.02), with increased risk for Medicaid and uninsured populations. Moreover, Medicaid PPS was also associated with greater adjusted lengths of stay and total hospital charges (p<0.0001). Importantly, these results were dependent on operation type. Primary payer status is associated with risk-adjusted postoperative mortality, morbidity, and resource utilization among pediatric surgical patients. Uninsured patients are at increased risk for postoperative mortality while Medicaid patients accrue greater morbidity, hospital lengths of stay, and total charges. These results highlight a complex interaction between socioeconomic and patient-related factors, and primary payer status should be considered in the preoperative risk stratification of pediatric patients.


Thrombin mutant W215A/E217A acts as a platelet GPIb antagonist.
Thrombin containing the mutations Trp215Ala and Glu217Ala (WE) selectively activates protein C and has potent antithrombotic effects in primates. The aim of this study was to delineate the molecular mechanism of direct WE-platelet interactions under static and shear conditions. Purified platelets under static conditions bound and spread on immobilized wild-type but not WE thrombin. In PPACK-anticoagulated blood under shear flow conditions, platelets tethered and rolled on both wild-type and WE thrombin, and these interactions were abrogated by the presence of a glycoprotein Ib (GPIb)-blocking antibody. Platelet deposition on collagen was blocked in the presence of WE, but not wild-type thrombin or prothrombin. WE also abrogated platelet tethering and rolling on immobilized von Willebrand factor in whole blood under shear flow. These observations demonstrate that the thrombin mutant WE, while not activating platelets, retains the ability to interact with platelets through GPIb, and inhibits GPIb-dependent binding to von Willebrand factor-collagen under shear.


Yeasts from autochthonal cheese starters: technological and functional properties.
The aim of this work was to identify 20 yeasts isolated from autochthonal cheese starters and evaluate their technological and functional properties. The capacities of the yeasts to grow at different temperatures, pH, NaCl and lactic acid concentrations as well as the proteolytic and lipolytic activities were studied. Moreover, survival to simulated gastrointestinal digestion, hydrophobicity, antimicrobial activity against pathogens and auto- and co-aggregation abilities were evaluated. The sequentiation of a fragment from the 26S rDNA gene indicated that Kluyveromyces marxianus was the predominant species, followed by Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces lactis and Galactomyces geotrichum. RAPD with primer M13 allowed a good differentiation among strains from the same species. All strains normally grew at pH 4.7-5.5 and temperatures between 15 and 35°C. Most of them tolerated 10% NaCl and 3% lactic acid. Some strains showed proteolytic (eight isolates) and/or lipolytic (four isolates) capacities. All strains evidenced high gastrointestinal resistance, moderate hydrophobicity, intermediate auto-aggregation and variable co-aggregation abilities. No strains inhibited the growth of the pathogens assayed. Some strains from dairy sources showed interesting functional and technological properties. This study has been the first contribution to the identification and characterization of yeasts isolated from autochthonal cheese starters in Argentina. Many strains could be proposed as potential candidates to be used as probiotics and/or as co-starters in cheese productions.


Combining dependent tests to compare the diagnostic accuracies--a non-parametric approach.
In this paper, we propose a non-parametric approach for comparing diagnostic accuracies in multi-reader receiver operating characteristic (ROC) studies. The approach constructs a test from each reader by extending the conventional non-parametric method and then combines all the individual test statistics to draw an overall conclusion on the relative accuracies of different diagnostic tests. The method can handle both continuous and ordinal data. Compared to the existing non-parametric methods, the method is robust and effectively deals with the possible heterogeneity among readers. It can also be applied to the analysis of correlated ROC studies. The method is applied to a real example and its finite sample performance is examined through simulation studies.


[Pathophysiology of complex regional pain syndrome (CRPS) type 1].
If the pathophysiology of complex regional pain syndrome (CRPS) type 1 remains controversial, most authors agree on a combination in varying proportions, a sensitization of peripheral nerves. To describe the state of advances in the physiopathology of complex regional pain syndrome type 1. Bibliographic research and literature review performed by referring to databases (Medline, Science Direct) The physiopathology of complex regional pain syndrome type 1 remains still poorly understood and controversial. Several arguments demonstrated both peripheral (inflammation, abnormal sympathetic ...) and central (neurological and cognitive) mechanisms. A better knowledge of the physiopathology of complex pain syndrome type 1 is necessary in order to adapt efficient curative therapy or to a better prevention of this syndrome.


Anatomic segmentectomy in the treatment of stage I non-small cell lung cancer.
Segmentectomy for early-stage non-small cell lung cancer (NSCLC) remains controversial and has been previously associated with high local recurrence rates. We compared the outcomes of anatomic segmentectomy with lobectomy for stage I NSCLC and investigated the impact of surgical resection margins on recurrence. From 2002 to 2006, 182 anatomic segmentectomies (114 open, 68 video-assisted thoracic surgery [VATS]), were performed for stage 1A (n = 109) or IB (n = 73) NSCLC. These were compared with 246 lobectomies (1A, 114; 1B, 132). Variables analyzed included hospital course, mortality, and patterns of recurrence and survival. All segmentectomy surgical margins were free of tumor (average margin, 18.2 mm). Operative time (147 versus 216 minutes; p < 0.0001) and estimated blood loss (185 versus 291 mL; p = 0.0003) were significantly reduced after segmentectomy compared with lobectomy. Thirty-day mortality (1.1% versus 3.3%), total complications, disease-free recurrence, and survival were similar between segmentectomy and lobectomy at a mean follow-up of 18.1 and 28.5 months, respectively. There were 32 recurrences after segmentectomy (17.6%) at a mean of 14.3 months (14 locoregional [7.7%], 18 distant [9.9%]), and 89% of recurrences were seen when tumor margins were 2 cm or less. Margin/tumor diameter ratios exceeding 1 were associated with a significant reduction in recurrence rates compared with ratios of less than 1 (25.0% versus 6.2%; p = 0.0014). Anatomic segmentectomy can be performed safely by an open or VATS approach. Segmentectomy outcomes compare favorably with standard lobectomy for stage I NSCLC. Margin/tumor ratios of less than 1 are associated with a higher rate of recurrence. Lobectomy should be considered as primary therapy when such margins are not obtainable with segmentectomy in the good-risk patient.


Immunotherapy for lung cancer combining the oligodeoxynucleotides of TLR9 agonist and TGF-β2 inhibitor.
Tumor immunotherapies have shown promising antitumor effects, especially immune checkpoint inhibitors (ICIs). However, only 12.46% of the patients benefit from the ICIs, the rest of them shows limited effects on ICIs or even accelerates the tumor progression due to the lack of the immune cell infiltration and activation in the tumor microenvironment (TME). In this study, we administrated a combination of Toll-like receptor 9 (TLR9) agonist CpG ODN and Transforming growth factor-β2 (TGF-β2) antisense oligodeoxynucleotide TIO3 to mice intraperitoneally once every other day for a total of four injections, and the first injection was 24 h after LLC cell inoculation. We found that the combination induced the formation of TME toward the enrichment and activation of CD8+ T cells and NK cells, accompanied with a marked decrease of TGF-β2. The combined therapy also effectively inhibited the tumor growth and prolonged the survival of the mice, even protected the tumor-free mice from the tumor re-challenge. Both of CpG ODN and TIO3 are indispensable, because replacing CpG ODN with TLR9 inhibitor CCT ODN showed no antitumor effect, CpG ODN or TIO3 alone did not lead to ideal antitumor results. This effect was possibly initiated by the activation of dendritic cells at the tumor site. This systemic antitumor immunotherapy with a combination of the two oligonucleotides (an immune stimulant and an immunosuppressive cytokine inhibitor) before the tumor formation may provide a novel strategy for clinical prevention of the postoperative tumor recurrence.


Inhibition of alpha-chymotrypsin by 2-halogeno-ethanols; comparisons with 2-methyl-ethanols and urea.
2-Halogeno- and 2-methyl-ethanols inhibit alpha-chymotrypsin in the order of their substituted groups: [1] tri- greater than di- greater than mono-, [2] Br- greater than Cl- greater than CH3- greater than F-. The inhibition by the halogeno-ethanols is mediated differently from that by the methyl-ethanols, ethanol, and urea.


Omega-3 polyunsaturated fatty acids in the treatment of atherogenic dyslipidemia.
Epidemiological studies have established an association between high triglycerides (TG) plasma levels and increased cardiovascular risk. Increased TG levels, commonly coupled with low HDL-C levels, are common in high cardiovascular risk subjects including those with dyslipidemia, metabolic syndrome and type 2 diabetes. Management of hypertriglyceridemia (HTG) includes lifestyle modification for mild-to-moderate HTG and pharmacological therapies for the treatment of high and very high TG levels. Among drugs, fibrates, nicotinic acid and omega-3 polyunsaturated fatty acids may be considered. Omega-3 fatty acids reduce plasma TG levels by several mechanisms; beside the effects on TG, omega-3 can also influence the levels of other lipids and lipoproteins including HDL-C and LDL-C. Clinical trials have also shown that omega-3 fatty acid supplementation is effective also when added in combination with other lipid-lowering drugs. These findings suggest that omega-3 fatty acids may be usefully considered for the management of high TG levels.


Severity of diverticulitis in patients with polycystic kidney disease without transplantation.
Patients with polycystic kidney disease (PKD) who have had a kidney transplant have an increased risk of diverticular disease and complicated diverticulitis. Literature is limited regarding the severity of diverticulitis in patients with PKD who have not had a transplant. We aim to assess whether patients with PKD, with and without renal transplant, have a similar course of diverticulitis. A retrospective review of all adult PKD patients at our institution diagnosed with diverticulitis between 2000 and 2016 was conducted. Patients without documented PKD and diverticulitis were excluded. We compared PKD patients with and without renal transplantation. A total of 41 patients were identified. Mean age was 60 (± 12), and 56% were female. Fourteen patients had undergone renal transplantation. Five (19%) non-transplant patients had complicated diverticulitis, compared to 43% (n = 6) transplanted (p = 0.33). Fifteen (56%) non-transplant and 8 (57%) transplant patients had recurrent diverticulitis (p = 1.00). Three (11%) non-transplant and 5 (36%) transplanted patients had recurrent complicated diverticulitis. Eight (30%) non-transplant and 7 (50%) transplant patients underwent surgery (p = 0.31). All 8 non-transplant patients underwent sigmoid resection with primary anastomosis without diversion. In the transplant group, 3 Hartmann procedures and 1 sigmoid resection with and 3 without diversion were performed. There was one in-hospital death in each group. In our group of patients, there was no difference in rate of recurrent diverticulitis, diverticulitis complications, or operative intervention in patients with PKD with and without renal transplant. The renal transplant group had a higher rate of recurrent, complicated diverticulitis.


The kinetics and thermodynamics of the reaction of solid-state fully reduced membrane-bound cytochrome oxidase with carbon monoxide as studied by dual-wavelength multichannel spectroscopy and flash photolysis.
1. The results of non-linear optimization studies on the mechanism of reaction of solid-state fully reduced membrane-bound cytochrome oxidase with CO over the 178--203 K range are presented. The analysis is carried out on data obtained by dual-wavelength multichannel spectroscopy at three wavelength pairs (444--463 nm, 590--630 nm and 608--630 nm), which yield three distinct progress curves. The only model that satisfies the triple requirement of a standard deviation within the standard error of the data, a random distribution of residuals and good determination of the optimized parameters is a two-species sequential mechanism: flash photolysis yields unliganded cytochrome oxidase and free CO, which then recombine to form species Ic; Ic is then converted into species IIc, which is identical with the cytochrome oxidase-CO complex existing before flash photolysis. All the thermodynamic parameters describing this model are calculated. 2. On the basis of the data obtained from this paper, together with data from potentiometric studies, magnetic susceptibility measurements and i.r. spectroscopy, the chemical identity of the species is suggested.


Reengineering the pharmaceutical industry by crash-testing molecules.
The recent decline in drug approvals and the increase in late-stage failures indicate that the ability to generate and screen large numbers of molecules has not improved the drug pipeline. Perhaps the pharmaceutical industry should follow the example of the automotive industry and agree upon a shared modeling language with vendors and academics to enable integration of predictive computational tools across the industry. This will then enable the virtual 'crash-testing' of drugs before synthesis, biological testing and, most importantly, clinical trials. This represents an ambitiously progressive approach using the models for simulating every stage of the drug discovery and development process. Combining the relevant computational algorithms into a grand unified model would enable prioritization of the best ideas before pursuing a discovery program, selecting a target or synthesizing a molecule. The successful application of these virtual crash-testing principles by any of its current proponents could revitalize the pharmaceutical industry so that failure is avoided.


Theileria equi merozoite antigen-2 interacts with actin molecule of equine erythrocyte during their asexual development.
Theileria equi is a tick-transmitted intraerythrocytic protozoan parasite in equids. Equine merozoite antigen (EMA)-1 and EMA-2 of T. equi have been identified as immunodominant proteins co-expressed on the surface of extra-erythrocytic merozoites. Additionally, only the EMA-2 is shed into the cytoplasm of infected erythrocyte or inside the erythrocytic membrane during their early developmental stage. In this study, we initially performed West-Western blot analysis on Triton X-100-insoluble erythrocytic skeleton collected from a healthy horse, using a glutathione S-transferase (GST)-tagged recombinant EMA-1t or EMA-2t of T. equi. The results indicated positive interactions of actin and band 4.1 molecules in the equine erythrocytic skeleton only with the recombinant EMA-2t. Subsequently, we carried out GST pull-down assay using the recombinant antigens (as above) against solubilized lysate of equine erythrocytic skeleton, and confirmed the co-precipitation of actin molecule with EMA-2t, but not with the EMA-1t. The interaction of EMA-2 with host erythrocytic actin indicated its role in the pathobiology of T. equi infection within host erythrocytes.


Atherosclerosis and inflammation.
The pathophysiology of coronary atherosclerosis is complex and multifactorial. The probability of the development of symptomatic coronary heart disease may be predicted by standard risk factor stratification involving hypertension, dyslipidemia, age, positive family history, and diabetes. However, risk factor stratification has been demonstrated to have significant limitations in the individual patient, which has generated a search for more specific and sensitive markers. Evidence is increasing that atherosclerosis is a disease characterized by inflammation, beginning with the earliest identifiable lesion (fatty streak) to the advanced vulnerable plaque. Clinical markers of inflammation, including C-reactive protein, modified low-density lipoprotein, homocysteine, tumor necrosis factor, and thermogenicity, have been identified as emerging risk factors that may add prognostic information in patient management. This review centers on inflammation as a potential pathogenetic factor in atherosclerosis and the role that clinical markers may play in the identification of patients at risk.


A method for addressing research gaps in HTA, developed whilst evaluating robotic-assisted surgery: a proposal.
When evaluating health technologies with insufficient scientific evidence, only innovative potentials can be assessed. A Regional policy initiative linking the governance of health innovations to the development of clinical research has been launched by the Region of Emilia Romagna Healthcare Authority. This program, aimed at enhancing the research capacity of health organizations, encourages the development of adoption plans that combine use in clinical practice along with experimental use producing better knowledge. Following the launch of this program we developed and propose a method that, by evaluating and ranking scientific uncertainty, identifies the moment (during the stages of the technology's development) where it would be sensible to invest in research resources and capacity to further its evaluation. The method was developed and tested during a research project evaluating robotic surgery. A multidisciplinary panel carried out a 5-step evaluation process: 1) definition of the technology's evidence profile and of all relevant clinical outcomes; 2) systematic review of scientific literature and outline of the uncertainty profile differentiating research results into steady, plausible, uncertain and unknown results; 3) definition of the acceptable level of uncertainty for investing research resources; 4) analysis of local context; 5) identification of clinical indications with promising clinical return. Outputs for each step of the evaluation process are: 1) evidence profile of the technology and systematic review; 2) uncertainty profile for each clinical indication; 3) exclusion of clinical indications not fulfilling the criteria of maximum acceptable risk; 4) mapping of local context; 5) recommendations for research.Outputs of the evaluation process for robotic surgery are described in the paper. This method attempts to rank levels of uncertainty in order to distinguish promising from hazardous clinical use and to outline a research course of action. Decision makers wishing to tie coverage policies to the development of scientific evidence could find this method a useful aid to the governance of innovations.


Factors affecting survival of tree seedlings in North Queensland rainforests.
Seedlings of six species of rainforest trees with widely constrasting ecology and seed morphology were transplanted at 3 weeks of age into tree-fall gaps and the shaded understoreys at two rainforest sites (Curtain Fig and Lamins Hill) on the Atherton Tableland, North Queensland, Australia. In each forest habitat, half of the transplanted seedlings were protected from vertebrates by means of wire cages, and survival was monitored over 16 months. The main objective was to estimate the extent to which independent variables (forest, habitat, protection from vertebrates, and species) contribute to explaining survival differences among the seedlings. Significant differences existed in the mortality among species, whether caged or uncaged. Seedlings unprotected from vertebrates suffered greater mortality (i.e. averaged over all species and forests, mortality > 75%), compared to the caged seedlings (mortality < 40%). Seedling of most species survived better in forest gaps, though the effects varied significantly among the six species. Survival of caged seedlings in Curtain Fig was significantly greater than in Lamins Hill, a pattern attributed to the partial deciduousness of the Curtain Fig forest. Because only few of the uncaged seedlings survived to 16 months, we focussed on the caged seedlings to examine the role of light at the forest habitats on survival. Relationship between seedling mortality and increasing light availability vary significantly among the six species. However across species, seedling mortality decreased linearly as photosynthetically active radiation (PAR) rose from 0.48 to 2.0%, after which an asymptote was reached and further increase in light (up to 8% PAR) did not influence survival significantly. Overall, temporal, species and the various environmental variables and their interactions explained about 60% of the variations in the seedling mortality data, with protection and species differences making the greatest contributions.


Continuous versus discontinuous drinking of an ethanol liquid diet in peripubertal rats: effect on 24-h variation of lymph node and splenic mitogenic responses and lymphocyte subset populations.
Excessive alcohol consumption continues to be a major public health problem, particularly in the adolescent and young adult populations. Generally, such a behavior tends to be confined to the weekends, to attain frequently binge drinking. This study in peripubertal male rats compares the effect of the discontinuous feeding of a liquid diet containing a moderate amount of ethanol (6.2% wt/vol) to that of continuous ethanol administration or a control diet, taking as end points the 24-h variations of plasma prolactin levels and mitogenic responses and lymphocyte subset populations in submaxillary lymph nodes and spleen. Animals received the ethanol liquid diet starting on day 35 of life, the diet being similar to that given to controls except for that maltose was isocalorically replaced by ethanol. Ethanol provided 36% of the total caloric content. Every week, the discontinuous ethanol group received the ethanol diet for 3 days and the control liquid diet for the remaining 4 days. After 4 weeks, rats were killed at six time intervals, beginning at 0900 h. A significant decrease of splenic cells' response to concanavalin A, and of lymph node and splenic cells' response to lipopolysaccharide was found in rats under the discontinuous ethanol regime, when compared with control- or ethanol-chronic rats. Under discontinuous ethanol feeding, mean values of lymph node and splenic CD8(+) and CD4(+)-CD8(+) cells decreased, whereas those of lymph node and splenic T cells, and splenic B cells, augmented. In rats chronically fed with ethanol, splenic mean levels of CD8(+) and CD4(+)-CD8(+) cells augmented. Both modalities of ethanol administration disrupted the 24 h variation in immune function seen in controls. Mean plasma prolactin levels increased by 3.6-fold and 8.5-fold in rats chronically or discontinuously fed with alcohol, respectively. The immune parameters examined in an additional group of rats fed regular chow and water ad libitum did not differ significantly from control liquid diet. The results support the view that the discontinuous drinking of a moderate amount of ethanol can be more harmful for the immune system than a continuous ethanol intake, presumably by inducing a greater stress as indicated by the augmented plasma prolactin levels observed.


[Long-term management of the stable coronary patient. The optimization of the medical treatment: a real objective].
Cardiovascular disease is one of the major causes of early morbidity and death in the developed world, and is becoming a serious public health concern in many developing countries. Over the last 30 years, in the USA and France, coronary angioplasty has become a standard treatment for stable angina, and this despite the recommendations of Learned Societies concerning the treatment of this condition. Today, 85 % of angioplasty procedures are performed on patients with stable angina. This study presents meta-analyses that compare medical treatment with angioplasty, and examine the impact of these strategies on more specific populations such as the elderly and post-myocardial infarction patients. To our minds, this synthesis seems to be of particular importance as the COURAGE study has rekindled the debate by showing that improvements in medical treatment and way of life reduced mortality and the recurrence of MI at five years, whereas there was no positive impact of an invasive strategy in any of the subgroups. Nevertheless, as a whole, studies on this subject underscore the value of angioplasty in the medium term for symptom relief in the case of ineffective medical treatment, notably during an acute coronary syndrome both in patients under medical treatment and in those who underwent invasive therapy at the initial phase.


Seabirds at risk around offshore oil platforms in the north-west Atlantic.
Seabirds aggregate around oil drilling platforms and rigs in above average numbers due to night lighting, flaring, food and other visual cues. Bird mortality has been documented due to impact on the structure, oiling and incineration by the flare. The environmental circumstances for offshore hydrocarbon development in North-west Atlantic are unique because of the harsh climate, cold waters and because enormous seabird concentrations inhabit and move through the Grand Banks in autumn (storm-petrels, Oceanodroma spp), winter (dovekies, Alle alle, murres, Uria spp), spring and summer (shearwaters, Puffinus spp). Many species are planktivorous and attracted to artificial light sources. Most of the seabirds in the region are long-distance migrants, and hydrocarbon development in the North-west Atlantic could affect both regional and global breeding populations. Regulators need to take responsibility for these circumstances. It is essential to implement comprehensive, independent arm's length monitoring of potential avian impacts of offshore hydrocarbon platforms in the North-west Atlantic. This should include quantifying and determining the nature, timing and extent of bird mortality caused by these structures. Based on existing evidence of potential impacts of offshore hydrocarbon platforms on seabirds, it is difficult to understand why this has not been, and is not being, systematically implemented.


The actions of 5-hydroxytryptamine on isolated skeletal muscle preparations.
The effects of 5-hydroxytryptamine (5HT) on the mouse phrenic nerve--hemidiaphragm and frog sartorius muscle preparations were studied using twitch tension and intracellular recording methods. Between 1--7.5 mM 5HT transiently augmented the twitch response of the diaphragm to indirect, but not direct, stimulation. Above 5 mM 5HT the augmentation was followed by a concentration-dependent twitch blockade. The blockade was more apparent in anticholinesterase-treated preparations, and was seen to a lesser extent in directly stimulated muscles. Intracellular recording from diaphragm muscles revealed that 5HT depressed the amplitudes of both miniature endplate potentials (m.e.p.p.s.) and endplate potentials (e.p.p.s.), this effect being more pronounced in the presence of neostigmine. 5HT also greatly prolonged the duration of the e.p.p.s. In the sartorius muscles, 1--5 mM 5HT slowed and depressed action potentials. It is suggested that 5HT facilitates transmission by an anticholinesterase action and inhibits by blockade of receptor ion channels.


Hazard of botulinum toxin type A injection into the detrusor in patients with an artificial urinary sphincter.
We performed a retrospective chart review of two patients with a previously implanted artificial urinary sphincter (AUS) who became incontinent after the injection of botulinum toxin type A into the detrusor. Two prepubertal males with a myelomeningocele, neuropathic bladder and urinary incontinence developed bladder hypertonicity. Treatment by botulinum toxin type A injection into the detrusor resulted in a loss of fluid from the balloon and recurrent incontinence. This method of treatment in patients with a previously implanted AUS should be avoided, as not only does it appear to be of only temporary benefit, but also as demonstrated by these cases it can damage the AUS system.


Fractionated stir bar sorptive extraction using conventional and solvent-assisted approaches for enhanced identification capabilities of aroma compounds in beverages.
For successful profiling of aroma carriers in food samples, a highly efficient extraction method is mandatory. A two-step stir bar sorptive extraction (SBSE) approach, namely fractionated SBSE (Fr-SBSE), was developed to improve both the organoleptic and the chemical identification of aroma compounds in beverages. Fr-SBSE consists of two multi-SBSE procedures (mSBSE) performed sequentially on the same sample. The first extraction consists of a conventional mSBSE using three polydimethylsiloxane (PDMS) stir bars (1stmSBSE). This is followed by a solvent-assisted mSBSE performed on the same sample using three solvent-swollen PDMS stir bars (2nd SA-mSBSE). The 1stmSBSE mainly extracts apolar/medium polar solutes with log Kow values >2, while the 2nd SA-mSBSE mainly extracts polar solutes with log Kow values <2. After this two-step fractionation procedure, either thermal desorption (TD) or liquid desorption - large volume injection (LD-LVI), followed by GC-MS is performed on each set of three stir bars. A real-life sample of roasted green tea was used for method development. The performance of the Fr-SBSE method is further illustrated with sensory evaluations and GC-MS analysis for a stout beer sample. Compared to an extraction procedure with SA-mSBSE only, Fr-SBSE including a 1stmSBSE and a 2nd SA-mSBSE reduced co-elution of aroma compounds in the chromatograms and was capable of providing improved mass spectral quality for identification of 17 additional compounds in roasted green tea, and 12 additional compounds in stout beer, respectively. Moreover, odor description and characterization were clearly improved.


Peripheral injury enhances central regeneration of primary sensory neurones.
The success of peripheral and fetal neural tissue in promoting outgrowth of axons from the adult mammalian central nervous system has tended to focus attention on local interactions between extending axons and their environment. The contribution to axon regeneration of biochemical and morphological changes in injured neurones is more difficult to evaluate. We report here that long spinal axons of primary sensory neurones are 100 times more likely to regenerate into peripheral nerve grafts if their peripheral axons are also cut. Regenerative behaviour at the axon tip seems to be strongly influenced by inducible events in the nerve cell.


Molecular characterization of carbapenemase producing Klebsiella pneumoniae strains by multiplex PCR and PFGE methods: The first K.pneumoniae isolates co-producing OXA-48/KPC and KPC/NDM in Turkey.
Carbapenems are frequently used in the treatment of multidrug-resistant infections caused by Klebsiella pneumoniae. The aim of the study is to definition and incidence of transferable carbapenemase genes of carbapenem resistant K. pneumoniae (CRKP) and to determine clonal relatedness of these strains in tertiary care hospital in Turkey. Identification of all 100 K. pneumoniae isolates and low sensitivity to any of the carbapenem group antibiotics were determined by Vitek-2 (BioMérieux, France). The frequency of carbapenemase genes (blaOXA-48, blaNDM, blaKPC, blaVIM,blaIMP) and extended spectrum beta-lactamase (ESBL) genes (blaCTX-M, blaSHV, blaTEM) which frequently detected in Turkey, have been investigated by multiplex polymerase chain reaction (PCR). Clonal relatedness was determined using Pulsed-field gel electrophoresis(PFGE). Ninety five isolates carried at least one of the carbapenemase genes (81.05% blaOXA-48, 38.9% blaNDM, 9.47% blaKPC,1.05% blaVIM). One isolate was carried the blaOXA-48+KPC and the two isolates were carried the blaKPC+NDM. PFGE demonstrated the presence of 24 pulse types and 63.09% of the isolates were in four main pulse types. This study demonstrated the incidence of blaNDM is beginning to reach endemic levels, in addition to blaOXA-48 found endemic in Turkey. To our knowledge, this is the first report of the co-production of these two genes (blaKPC + NDM and blaOXA-48 + KPC) in CRKP isolates.


Pituitary adenylate cyclase activating polypeptide (PACAP) is localized in human dermal neurons and causes histamine release from skin mast cells.
Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide homologous with vasoactive intestinal polypeptide (VIP) which is known to induce histamine release in human skin mast cells. PACAP has not been detected in human skin. The purposes of the study were to investigate the occurrence of PACAP in human skin and to evaluate the histamine releasing activity of the two common pro-PACAP products, PACAP-27 and PACAP-38. Fourteen human surgical skin samples were obtained. PACAP and VIP were visualized by immunohistochemistry. A microdialysis technique was used to measure histamine release in intact skin samples following intradermal injections of the peptides. PACAP and VIP were localized in dermal nerves in connection with sweat glands. Intradermal injection of 3 or 10 microm PACAP significantly released histamine. Kinetics of histamine release showed peak release 2-4 min after skin challenge. Ten microm of PACAP-27, VIP and somatostatin caused histamine release with similar efficacy, whereas PACAP-38 was less effective. Substance P was twice as efficient as PACAP-27, whereas calcitonin gene-related peptide did not release histamine. PACAP is found in human skin and is capable of releasing histamine from skin mast cells.


Serological follow-up after non-typhoid salmonella infection in humans using a mixed lipopolysaccharide ELISA.
In industrialized countries, non-typhoid salmonella are a frequent cause of bacterial gastroenteritis. Diagnosis is usually made by stool culture, which is labour-intensive and time-consuming. Sensitivity depends on handling of stool samples and delay from illness onset to sampling. We developed an indirect mixed enzyme-linked immunosorbent assay (ELISA) for the detection of human serum antibodies against lipopolysaccharide antigens of the two predominant serovars, Salmonella serovar Enteritidis (S. Enteritidis) and S. Typhimurium. We measured IgA, IgM and IgG in 964 sera from 302 patients with stool culture-confirmed acute salmonella gastroenteritis, in 300 sera from healthy blood donors, and in 147 sera from patients with antibodies against other bacteria. Patient sera were collected within 1 month and approximately 3, 6, and 12 months after illness onset. For sera collected ≤ 30 days of onset, sensitivity of the ELISA was 92% for S. Enteritidis and 86% for S. Typhimurium, with a specificity of 95% for both serovars. The mixed ELISA is a useful additional tool for the diagnosis of acute salmonella gastroenteritis. It allows rapid analysis of multiple samples, thus can be used for sero-epidemiological studies of large population-based serum collections in order to estimate the population incidence of salmonella infections. Another application is aetiological diagnostics in patients with suspected post-infectious complications such as reactive arthritis, even when faecal shedding of salmonella has ceased.


Xanthomatosis and other clinical findings in patients with elevated levels of very low density lipoproteins.
Forty-six patients with xanthomatosis and elevated very low density lipoproteins (VLDL) levels (in different types of hyperlipoproteinaemia) were classified on the basis of the WHO criteria and the cholesterol/triglyceride ratio in VLDL. A large majority (31/46) of the patients referred to the Department of Dermatology could be classified as hyperlipoproteinaemia type III, only 8/46 as type IIB and 7/46 as type IV/V. This distinction seems to be relevant as the xanthomatous lesions differed distinctly between these three types of hyperlipoproteinaemia. Xanthochromia striata palmaris was present in 29/31 cases of hyperlipoproteinaemia type III and was not found in type IV/V patients, who had distinctive papuloeruptive xanthomas. During a follow-up in 35/46 patients all xanthomas disappeared within 2 years except the xanthelasma palpebrarum and tendinous xanthomas. All type IV/V patients (7/7) but only one type III patient (1/31) had abnormal glucose tolerance. Only 2/18 type III patients less than 45 years showed claudication and none of the young type III patients had angina pectoris. In contrast, all four type IIB patients less than 45 years had clinical signs of atherosclerosis. However, angina pectoris and/or claudication were present in 5/13 type III patients over 45 years old. The mean serum cholesterol level was equally elevated in both groups but the cholesterol was mainly present in VLDL in type III and in low density lipoproteins (LDL) in type IIB. In 9/31 type III patients the LDL level was also elevated but was easily normalized by a diet low in carbohydrate, whereas the elevated LDL level in type IIB was therapy-resistant. The recognition of xanthomatous lesions, specifically xanthochromia striata palmaris, as an early sign of type III hyperlipoproteinaemia, can lead to the early diagnosis and successful treatment of these patients, and thus possibly prevent the development of premature atherosclerosis.


Inhibition of N-methyl-D-aspartate evoked sodium flux by MK-801.
The inhibition of N-methyl-D-aspartate (NMDA) stimulated 22Na+ efflux from rat hippocampal slices was studied using competitive and non-competitive receptor antagonists. There was a good correlation between the abilities of the competitive antagonists to block NMDA evoked 22Na+ efflux and their potencies as inhibitors of L-[3H]glutamate binding. The recently reported novel NMDA receptor antagonist, (+)-5-methyl-16,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801) was shown to non-competitively inhibit NMDA stimulated 22Na+ efflux with an IC50 value of 0.4 microM. Relatively high (10 microM) concentrations of MK-801 had no effects on quisqualic acid, alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA), or kainic acid stimulated efflux. However, MK-801 was able to block 22Na+ efflux induced by ibotenic acid and L-homocysteic acid, amino acids that act as NMDA receptor agonists. MK-801, (-)-MK-801, and non-competitive NMDA receptor antagonists of the arylcyclohexylamine and dioxolane classes inhibited NMDA stimulated 22Na+ efflux with potencies that reflected their abilities to compete for [3H]MK-801 binding sites in rat cortical membranes. These results indicate the utility of the 22Na+ efflux assay in studying the properties of NMDA receptors and confirm the nature and selectivity of the inhibition of NMDA receptor linked ion channel activation by MK-801.


Iatrogenic lithotripsy failure: penetration of shock waves through tape.
Foam tape used to protect epidural catheters during immersion in a Dornier HM3 lithotriptor was identified retrospectively as the cause of poor stone fragmentation. Studies of shock wave penetration through various protective type materials indicate that a particular water repellant tape is least likely to impair shock wave penetration.


Effect of harvest time span on physicochemical properties, antioxidant, antimicrobial, and anti-inflammatory activities of Meliponinae honey.
The maturity of honey has a great impact on its quality and contents. Additionally, stingless bee honey contains high moisture, which allows microorganisms to survive and ferment, contributing to honey's variable flavor and physicochemical properties. Therefore, there is a need for better quality control of the honey process, especially the harvest time of honey. We gathered honey from the nest of stingless bees Heterotrigona itama and Tetrigona binghami over different time periods, i.e. 15, 30, and 45 days. The results show harvest time considerably affects the physicochemical properties, antioxidant activity, and antimicrobial activity of honey. Good antioxidant activity and antimicrobial activity can be found in honey produced from a longer harvest time. Compared with 15-day harvest time, at 30- or 45-day harvest time water, trehalulose, and protein content and total acidity increased, and the content of reducing sugars, fructose and glucose, and pH values, decreased in both types of honey. Moreover, compared with 15-day harvest time, the sum of six organic acids in the 45-day honey of H. itama fluctuated between 2.78 to 4.12 g 100 g-1 and in the 45-day honey of T. binghami increased from 1.66 to 3.61 g 100 g-1 , respectively. Honey harvest time had a significant effect on the physicochemical properties, antioxidant activity, and antimicrobial activity of stingless bee honey (H. itama or T. binghami). This study provides a reference for beekeepers to adjust harvest time to obtain honey with suitable physicochemical parameters. © 2022 Society of Chemical Industry.


Carbon and hydrogen isotope ratios of bryophytes from arid and humid regions.
Measurements of carbon isotope ratios of nine taxa of arid zone mosses from North America and 24 taxa of European bryophytes show consistent values indicative of C3 metabolism. Other morphological, physiological and biochemical characteristics provide the mechanism of adaptation to environmental stresses of heat and extended desiccation in these plants. Analyses of D content indicate that values become less negative with increasing aridity of the habitat.


Mycoplasma hominis as a respiratory pathogen in X-linked hypogammaglobulinaemia.
We report a case of lower respiratory tract infection with Mycoplasma hominis in a patient with X-linked hypogammaglobulinaemia.


Nursing diagnosis: ineffective coping. II. Planning care.
It has been the intent of this discussion to provide nurses with a model for understanding the coping process and a framework for planning interventions that can affect the stress-coping cycle in a variety of ways. The model also provides a framework for research that identifies effective nursing activities.


Phosphorylation Changes in Response to Kinase Inhibitor H89 in PKA-Null Cells.
Protein phosphorylation, mediated by protein kinases, plays a crucial role in cellular regulation. One of the most important protein kinases is protein kinase A (PKA). N-[2-p-bromocinnamylamino-ethyl]-5-isoquinolinesulphonamide (H89) is often used as a "PKA specific inhibitor" to study the involvement of PKA in signaling pathways. However, evidence from cell-free experiments has suggested that H89 can also inhibit other protein kinases. In this study, previously generated PKA-null and PKA-intact mouse cell lines derived from mpkCCD cells were treated with H89 over a range of concentrations commonly used in the literature, followed by mass spectrometry-based phosphoproteomics to globally assess changes in phosphorylation. From a total of 14,139 phosphorylation sites quantified, we found that 571 and 263 phosphorylation sites with significant changes in abundance in PKA-intact and PKA-null cells, respectively. Analyses of sequence logos generated from significantly decreased phosphorylation sites in PKA-intact and PKA-null cells both revealed a preference for basic amino acids at position -3 and -2. Thus, H89 appears to inhibit basophilic kinases even in the absence of PKA. Likely H89 targets include basophilic protein kinases such as AKT, RSK, AMPK and ROCK. We conclude that, in intact cells, H89 can affect activities of protein kinases other than PKA, and therefore responses to H89 should not be regarded as sufficient evidence for PKA involvement in a signaling process.


[Detection of herpes simplex virus 2 (HSV) IgG and IgM using ELISA in transsexuals and homosexuals].
In this study, the sera of 12 cases of transsexuals with vaginoplasty and 15 cases of homosexuals have been investigated with ELISA from the view point of HSV-2 IgG and IgM. Twenty-five cases were within the 18-26 age groups and the other 2 transsexuals were 36 years old. Our study group has not significant clinical pathology. All of the 27 cases of transsexuals and homosexuals have been found positive for HSV-2 IgG. In 7 cases HSV-2 IgM have been found positive, the other 20 cases gave negative results. In the one third of the transsexuals and one fifth of the homosexuals, HSV-2 IgG have been found positive. Twenty-six percent of seropositivity in transsexuals and homosexuals show that they are the significant risk group for HSV-2 infection.


Quantitative analysis of methylation of genomic loci in early-stage rectal cancer predicts distant recurrence.
There are no accurate prognostic biomarkers specific for rectal cancer. Epigenetic aberrations, in the form of DNA methylation, accumulate early during rectal tumor formation. In a preliminary study, we investigated absolute quantitative methylation changes associated with tumor progression of rectal tissue at multiple genomic methylated-in-tumor (MINT) loci sequences. We then explored in a different clinical patient group whether these epigenetic changes could be correlated with clinical outcome. Absolute quantitative assessment of methylated alleles was used to assay methylation changes at MINT 1, 2, 3, 12, 17, 25, and 31 in sets of normal, adenomatous, and malignant tissues from 46 patients with rectal cancer. Methylation levels of these biomarkers were then assessed in operative specimens of 251 patients who underwent total mesorectal excision (TME) without neoadjuvant radiotherapy in a multicenter clinical trial. Methylation at MINT 2, 3, and 31 increased 11-fold (P = .005), 15-fold (P < .001), and two-fold (P = .02), respectively, during adenomatous transformation in normal rectal epithelium. Unsupervised grouping analyses of quantitative MINT methylation data of TME trial patients demonstrated two prognostic subclasses. In multivariate analysis of node-negative patients, this subclassification was the only predictor for distant recurrence (hazard ratio [HR], 4.17; 95% CI, 1.72 to 10.10; P = .002), cancer-specific survival (HR, 3.74; 95% CI, 1.4 to 9.43; P = .003), and overall survival (HR, 2.68; 95% CI, 1.41 to 5.11; P = .005). Methylation levels of specific MINT loci can be used as prognostic variables in patients with American Joint Committee on Cancer stage I and II rectal cancer. Quantitative epigenetic classification of rectal cancer merits evaluation as a stratification factor for adjuvant treatment in early disease.


Autophagy promotes degradation of internalized collagen and regulates distribution of focal adhesions to suppress cell adhesion.
Adhesion of cells to the extracellular matrix (ECM) via focal adhesions (FAs) is crucial for cell survival, migration, and differentiation. Although the regulation of FAs, including by integrins and the ECM, is important to cell behavior, how FAs are regulated is not well known. Autophagy is induced by both cell adhesion and cell detachment. Here, we showed that autophagosomes are located close to internalized collagen and paxillin, which is a well-known marker of FAs. Autophagy-deficient cells showed increased levels of internalized collagen compared with control cells. Moreover, paxillin exhibited a more peripheral distribution and the area of paxillin was increased, and adhesion-induced focal adhesion kinase signaling was impaired and adhesion was enhanced, in autophagy-deficient cells. These results suggest that autophagy suppressed cell adhesion by regulating internalized ECM and FAs.


Revisiting the Predictive Factors for Nodal Metastasis in Oral Cancers-a Leap Towards Personalized Patient Treatment Approach.
The incidence of oral squamous cell carcinoma is steadily increasing globally, and even with a better understanding of tumor biology and advanced treatment modalities, the survival of OSCC patients is still not improved. A single metastatic cervical node can decrease survival by 50%. Our study intends to identify the clinical, radiological, and histological factors, significant for nodal metastasis in the pretreatment setting. Ninety-three patients' data is prospectively collected and analyzed to identify the significance of various factors in predicting nodal metastasis. Clinical factors like smokeless tobacco and nodal characteristics and T category and radiological factors like the number of specific nodes were significant for pathological nodes on univariate analysis. Ankyloglossia, radiological ENE, and radiological nodal size were significant on multivariate analysis also. In the pretreatment setting, clinicopathological and radiological factors can be used to predict nodal metastasis in generating predictive nomograms and for better planning of treatment.


Influence of three scan spray systems on human gingival fibroblasts.
CAD/CAM is based on optical or mechanical scanning of tooth surfaces. The aim of this in vitro study was to investigate the influence of three different scan sprays on the proliferation, viability, and adenylate kinase (ADK) release of human fibroblasts. Three different scan sprays (ScanDry, ScanSpray Luer Classic, and CEREC Optispray) were tested in vitro to determine their effects on proliferation, viability, and ADK release of human periodontal fibroblasts. A defined amount of the test material was sprayed into 96 multiwell plates, dried, and incubated with fibroblasts. The LIVE/DEAD Viability Assay, a two-color fluorescence-based method, was used to determine the cytotoxic potential (the AlamarBlue Assay for the proliferation rate and the ToxiLight BioAssay for the release of ADK). There were differences between the scan sprays concerning inhibition of the proliferation and viability of fibroblasts. All materials inhibited the fibroblast proliferation and viability compared with the control group (P < .001), and there were also significant differences among the scan spray groups. The scan sprays led to a greater release of ADK, but a significant difference could be found only between ScanSpray Luer Classic and CEREC Optispray (P < .009). The results of this study proved that the scan sprays do not induce a significantly higher ADK release than the control group. The inhibiting effect on the fibroblast proliferation can be attributed to a material-independent phenomenon. Further experiments are necessary to validate the present data.


A comparison of different photosensitizing dyes with respect to uptake C3H-tumors and tissues of mice.
Nine dyes, all potential sensitizers for photodynamic cancer therapy (PDT), were injected in mice with C3H mammary carcinomas. Twenty-four hours later the animals were sacrificed and the dye concentrations in tumors and 9 other tissues were measured by means of spectrofluorimetry. The 9 dyes were: Photofrin II (PII), hematoporphyrin (HP)-di-hexyl-ether, PSD-007 (a sensitizer used in clinical trials in China), tetraphenyl porphine tetrasulfonate (TPPS4), tetra(3-hydroxy phenyl)porphyrin (3THPP), aluminium phthalocyanine tetrasulfonate (AlPCTS), aluminium phthalocyanine (AlPC), chlorin e6 (Chl e6) and merocyanine 540 (MC 540). The porphyrin precursor delta-aminolevulinic acid was also tested and found to induce porphyrin fluorescence in tumors and some other tissues. The best tumorlocalizer of those tested was 3THPP. This drug also showed a favorable tissue distribution. The following dyes showed lower skin/tumor concentration ratios than PII (the most widely used dye for PDT): Chl e6, PSD-007, HP-di-hexyl-ether and 3THPP. Low brain/tumor ratios were found for: PSD-007, HP-di-hexyl-ether, 3THPP, TPPS4 and AlPCTS.


Relationship between anxiety and quality of life among older adults with self-reported polypharmacy in long-term care: A cross-sectional study.
To investigate the relationship between anxiety and quality of life among older adults with self-reported polypharmacy living in the long-term care setting. A cross-sectional design was used. Between July 2021 and August 2022, 92 older adults living in long-term care completed an anonymous one-time questionnaire packet. Polypharmacy was measured as self-reported five or more medications daily. Anxiety was measured using the Geriatric Anxiety Scale-Long Term Care tool. Quality of life was measured as health-related quality of life using two global questions from the RAND-36 and as medication-related quality of life using the Medication-Related Quality of Life Scale. The prevalence of polypharmacy was 89%. Among participants with polypharmacy, average age was 80.1 ± 7.9 years. The majority were female (70%) and white (85%). There was a moderate-to-strong correlation between anxiety and quality of life. Specifically, anxiety was negatively related to current health-related quality of life, perceived change in health-related quality of life and medication-related quality of life. Overall, anxiety explained 27-31% of the variance in both health-related and medication-related quality of life. The results of this study indicate that as anxiety increases, health-related and medication-related quality of life decreases in older adults living in long-term care who report consuming five or more medications daily. Advanced practice nurses can use these findings to guide practice, tailor interventions and improve care for these long-term care residents. Multiple medications are increasingly prescribed to treat multiple comorbidities in older adults. As a result, the prevalence of polypharmacy (≥5 medications per day) is rising and problematic. The main findings of this study highlight the negative relationship between anxiety and quality of life in this population and the need for adequate assessment of anxiety by advanced practice nurses in order to personalize care. In preparing the manuscript, the authors have adhered to relevant EQUATOR guidelines and the STROBE checklist for cross-sectional studies. No patient or public contribution outside of participation in the actual study for purposes of data collection.


Tikhonov regularization of a second order dynamical system with Hessian driven damping.
We investigate the asymptotic properties of the trajectories generated by a second-order dynamical system with Hessian driven damping and a Tikhonov regularization term in connection with the minimization of a smooth convex function in Hilbert spaces. We obtain fast convergence results for the function values along the trajectories. The Tikhonov regularization term enables the derivation of strong convergence results of the trajectory to the minimizer of the objective function of minimum norm.


Maternal Knowledge, Attitude and Practices toward Free Sugar and the Associations with Free Sugar Intake in Children.
Research addressing factors related to free sugar (FS) consumption among children in Saudi Arabia is lacking. We aimed to evaluate maternal knowledge, attitude, and practices toward FS and the associations with children's intake of FS. This cross-sectional study included 424 Saudi children aged 6-12 years and their mothers. Data related to maternal knowledge, attitude, and practices were collected using an online survey. Data concerning children's habitual intake of FS were collected through phone interviews using a validated food frequency questionnaire. Limited knowledge on FS was observed among mothers of children [median 7.00 [interquartile range 6.00-8.00] out of 11.0. Maternal knowledge was not correlated with maternal attitude or practices toward FS. Maternal knowledge towards FS did not predict children's intake of FS, whereas maternal attitude and practices toward limiting the consumption of FS predicted lower intake of FS among Saudi children, particularly the FS consumed from solid food sources (B: -5.73 [95% confidence interval (CI): -9.79 to -1.66]) and (B: -6.85 [95% CI: -11.9 to -1.80]), respectively. Despite the limited knowledge pertaining to FS among mothers in Saudi Arabia, they were making efforts to limit their children's consumption of FS.


A fiber optic biosensor (FOBS) to monitor mutans streptococci in human saliva.
A fiber optic biosensor (FOBS) to monitor mutans streptococci activity in human saliva is developed. The biosensor utilizes e fiber optic evanescent wave spectroscopy to monitor a bacterial mediated biochemical reaction. To achieve this, a short length of the cladding is removed; the fiber core surface is treated and coated with a thin film of porous glass medium using sol-gel technique. The mutans streptococci mediated reaction with sucrose is monitored using a photosensitive indicator, which is immobilized within the porous glass coating. Spectroscopic analysis shows that the transmitted intensity at 597 nm increases conspicuously when monitored for 120 min. Two distinct phases are observed, one from 0 to 60 min and the other from 60 to 120 min. A negative correlation coefficient between the rate of increase in absorption peak intensity recorded by the FOBS and the decrease in pH measured using the pH meter, was calculated to be rho=-0.994. This investigation highlights the potential benefits of this sensor to monitor mutans streptococci activity in saliva.


The high prevalence of HLA-B5 in Behçet's disease.
Sixteen of the nineteen (84%) patients with Behçet's disease in Turkey had HLA-B5; the prevalence of HLA-B5 among 150 controls was 27%.


Dopaminergic effects on electrophysiological and functional MRI measures of human cortical stimulus-response power laws.
Power laws have been widely used to formulate relationships between objective intensity of stimulation and subjective intensity of sensation. We investigated the effects of dopaminergic drug treatment (sulpiride) on the relationship between somatosensory stimulus intensity and cortical response measured electrophysiologically by somatosensory-evoked potentials (SEP) and functional magnetic resonance imaging (fMRI). The intensity of stimulation was related by a simple power law to both electrophysiological and fMRI measures of cortical response, with overlapping confidence intervals for both power law exponents. Sulpiride did not modulate the power law exponent, but significantly attenuated the "gain" of both stimulus-response functions. Using path analysis we decomposed dopaminergic effects on fMRI data into an indirect component (16%), predictable by drug effects on SEP, and a direct component (84%), not explained electrophysiologically. Results indicate that sulpiride has comparable effects on power law parameters estimated from SEP and fMRI, but fMRI has superior sensitivity to detect drug effects on somatosensory cortical recruitment by graded stimulation.


Glibenclamide inhibits agonist-induced vasoconstriction of placental chorionic plate arteries.
Preliminary data suggest that K(ATP) channels may be expressed in placental arteries and veins [Wareing M, Turner C, Greenwood SL, Baker PN, Fyfe GK. Expression of mRNA encoding K+ channels in chorionic plate arteries and veins. J Soc Gynecol Investig 2004;11:353A]. However, no data exist on glibenclamide's effects in placental chorionic plate arteries. To assess the effect of glibenclamide on placental chorionic plate arterial vasoconstriction. Arteries were dissected from placental chorionic plate biopsies obtained at term from uncomplicated pregnancies (N=63). Arteries were mounted onto a wire myograph in HCO3- -buffered physiological salt solution (PSS) at 37 degrees C (5% O2/5% CO2 bubbling) and normalised at 0.9 of L5.1 kPa. Constriction viability was assessed with 120 mmol l(-1) potassium solution (KPSS). Dose-response curves were produced with the thromboxane-mimetics U46619 and U44069 (10(-10)-2 x 10(-6)M), arginine vasopressin (10(-10)-5 x 10(-8)M) and endothelin-1 (10(-11)-3 x 10(-7)M) in the presence or absence of 50 micromol l(-1) glibenclamide. The effect of glibenclamide on arginine vasopressin- and U46619-induced constriction was also assessed in the presence of the cyclo-oxygenase inhibitor indomethacin (10 micromol l(-1)). Pre-incubation with 50 micromol l(-1) glibenclamide significantly right-shifted dose-response curves to all vasoconstrictive agonists tested (repeated measures ANOVA). Indomethacin did not modify the inhibitory effect of glibenclamide. Glibenclamide's effects on agonist-induced constrictions are unlikely to be via an inhibition of ATP-sensitive K+ channels, and with U46619- and U44069-induced constrictions, glibenclamide may be acting as a competitive antagonist of thromboxane receptors.


Some approximation results for mild solutions of stochastic fractional order evolution equations driven by Gaussian noise.
We investigate the quality of space approximation of a class of stochastic integral equations of convolution type with Gaussian noise. Such equations arise, for example, when considering mild solutions of stochastic fractional order partial differential equations but also when considering mild solutions of classical stochastic partial differential equations. The key requirement for the equations is a smoothing property of the deterministic evolution operator which is typical in parabolic type problems. We show that if one has access to nonsmooth data estimates for the deterministic error operator together with its derivative of a space discretization procedure, then one obtains error estimates in pathwise Hölder norms with rates that can be read off the deterministic error rates. We illustrate the main result by considering a class of stochastic fractional order partial differential equations and space approximations performed by spectral Galerkin methods and finite elements. We also improve an existing result on the stochastic heat equation.


Evaluation of renal masses by ultrasound and computed tomography: a preliminary study.
Computed tomography (CT) and ultrasound were performed on fifty-two patients with renal masses. Both modalities were able to correctly demonstrate the renal mass with an equal degree of accuracy. Since renal ultrasound is a well established, economical and noninvasive technique, it should be the diagnostic procedure of choice to determine whether or not a mass demonstrated by intravenous urography is cystic or solid. However, if faster scanning times as well as lower patient cost could be accomplished, more widespread use of CT as a diagnostic modality for differentiation of renal masses could result.


Modulation of the exocellular serine-thiol proteinase activity of Paracoccidioides brasiliensis by neutral polysaccharides.
Our group characterized an exocellular serine-thiol proteinase activity in the yeast phase of Paracoccidioides brasiliensis (PbST), a dimorphic human pathogen. The fungal proteinase is able to cleave in vitro, at pH 7.4, proteins associated with the basal membrane, such as human laminin and fibronectin, type IV collagen and proteoglycans. In the present study, we investigated the influence of glycosaminoglycans (GAGs) and neutral polysaccharides upon the serine-thiol proteinase activity by means of kinetic analysis monitored with fluorescence resonance energy transfer (FRET) peptides using the substrate Abz-MKALTLQ-EDDnp (Abz=ortho-aminobenzoic acid; EDDnp=ethylenediaminedinitrophenyl). Only neutral polysaccharides exhibited patterns of interaction with the proteinase, while sulfated GAGs had no effect. Incubation with neutral polysaccharides resulted in a powerful modulation of the enzyme activity, intensely changing the enzyme kinetic parameters of catalysis and affinity for the substrate. Commercial dextran at the highest concentration of 20 microM increased 6.8-fold the enzyme affinity for the substrate. In the presence of 8 microM of purified baker's yeast mannan, the apparent KM of the enzyme increased about 5.5-fold, reflecting a significant inhibition in binding to the peptide substrate. When an exocellular galactomannan (GalMan) complex isolated from P. brasiliensis was added to the reaction mixture at 400 nM, the apparent KM and VMAX decreased about threefold. Moreover, GalMan was able to protect the enzymatic activity at high temperatures, but it caused no effect on the optimum cleavage pH. Our results show a novel modulation mechanism in P. brasiliensis, where a fungal polysaccharide-rich component can stabilize a serine-thiol proteolytic activity, which is possibly involved in fungal dissemination.


Imaging and quantitation of the hypoxic cell fraction of viable tumor in an animal model of intracerebral high grade glioma using [18F]fluoromisonidazole (FMISO).
We have demonstrated that FMISO uptake is significantly higher in tumor tissue in the C6 intracerebral glioma rat model compared to normal brain, and that there is persisting hypoxia in gliomas independent of tumor size. FMISO uptake was observed homogeneously throughout viable glioma tissue in tumor sizes ranging from 2mm to almost 1cm. Quantitation of uptake of FMISO showed a tumor/brain ratio of 1.9 and a tumor/blood ratio of 2.6 at 2 hours post injection.


[An analysis of the importance of low-molecular peptides in regulating human motor functions in diseases of the central nervous system].
The clinical data obtained on the treatment of motor disorders with intrathecal administration of low-molecular peptides, suggest that the peptides are effective in Parkinsonian disease, extrapyramidal hyperkineses and in spinal cord disturbances.


Harnessing SLE Autoantibodies for Intracellular Delivery of Biologic Therapeutics.
Intracellular delivery of therapeutic antibodies is highly desirable but remains a challenge for biomedical research and the pharmaceutical industry. Approximately two-thirds of disease-associated targets are found inside the cell. Difficulty blocking these targets with available drugs creates a need for technology to deliver highly specific therapeutic antibodies intracellularly. Historically, antibodies have not been believed to traverse the cell membrane and neutralize intracellular targets. Emerging evidence has revealed that anti-DNA autoantibodies found in systemic lupus erythematosus (SLE) patients can penetrate inside the cell. Harnessing this technology has the potential to accelerate the development of drugs against intracellular targets. Here, we dissect the mechanisms of the intracellular localization of SLE antibodies and discuss how to apply these insights to engineer successful cell-penetrating antibody drugs.


Evidence for the formation of a quinone methide during the oxidation of the insect cuticular sclerotizing precursor 1,2-dehydro-N-acetyldopamine.
1,2-Dehydro-N-acetyldopamine (dehydro-NADA) is an important catecholamine derivative involved in the cross-linking of insect cuticular components during sclerotization. Since sclerotization is a vital process for the survival of insects, and is closely related to melanogenesis, it is of interest to unravel the chemical mechanisms participating in this process. The present paper reports on the mechanism by which dehydro-NADA is oxidatively activated to form reactive intermediate(s) as revealed by pulse radiolysis, electron spin resonance spectroscopy, high performance liquid chromatography, and ultraviolet-visible spectroscopic analysis. Pulse radiolytic one-electron oxidation of dehydro-NADA by N3. (k = 5.3 x 10(9) M-1 s-1) or Br2.- (k = 7.5 x 10(8) M-1 s-1) at pH6 resulted in the rapid generation of the corresponding semiquinone radical, lambda max 400 nm, epsilon = 20,700 M-1 cm-1. This semiquinone decayed to form a second transient intermediate, lambda max 485 nm, epsilon = 8000 M-1 cm-1, via a second order disproportionation process, k = 6.2 x 10(8) M-1 s-1. At pH 6 in the presence of azide, the first order decay of this second intermediate occurred over milliseconds; the rate decreases at higher pH. At pH 6 in the presence of bromide, the intermediate decayed much more slowly over seconds, k = 0.15 s-1. Under such conditions, the dependence of the first order decay constant upon parent dehydro-NADA concentration led to a second order rate constant of 8.5 x 10(2) M-1 s-1 for reaction of the intermediate with the parent, probably to form benzodioxan "dimers." (The term dimer is used for convenience; the products are strictly bisdehydrodimers of dehydro-NADA (see "Discussion" and Fig. 11)) Rate constants of 5.9 x 10(5), 4.5 x 10(5), 2.8 x 10(4) and 3.5 x 10(4) M-1 s-1 were also obtained for decay of the second intermediate in the presence of cysteine, cysteamine, o-phenylenediamine, and p-aminophenol, respectively. By comparison with the UV-visible spectroscopic properties of the two-electron oxidized species derived from dehydro-NADA and from 1,2-dehydro-N-acetyldopa methyl ester, it is concluded that the transient intermediate exhibiting absorbance at 485 nm is the quinone methide tautomer of the o-quinone of dehydro-NADA. Sclerotization of insect cuticle is discussed in the light of these findings.


Expression pattern of asialoglycoprotein receptor in human testis.
During acute or chronic hepatitis B virus (HBV) infection, the virus can invade the male reproductive system, pass through the blood-testis barrier and integrate into the germ line, resulting in abnormal spermatozoa. However, the pathway remains unclear. The asialoglycoprotein receptor (ASGR), a potential receptor for HBV, is mainly distributed in hepatocytes. We have examined the distribution of ASGR in human testis and found it in the seminiferous tubules and interstitial region but its enrichment in human testis is much lower than that in liver. By multiple immunoenzyme histochemistry staining, ASGR was precisely co-localized with vimentin (Sertoli cell marker) but not proliferating cell nuclear antigen (spermatogonial cell marker) in testis tissue. ASGR was expressed in human Leydig cells, stromal cells in the seminiferous tubules and Sertoli cells but seldom in spermatogonial cells. Therefore, ASGR could provide HBV with access to the luminal compartment of human testis. The mechanism by which HBV invades germ cells remains unknown.


Conductive oxygen barrier films using supramolecular assembly of graphene embedded polyelectrolyte multilayers.
The supramolecular self-assembly of polyelectrolyte multilayers (PEMs) provides robust bottom-up strategies to assemble a broad spectrum of nanostructures on the host substrates. In this study, we discuss the formation of graphene nanoplatelet (GNP) embedded polyelectrolyte films to enhance the oxygen barrier properties of poly(ethylene terephthalate) (PET) films. Despite cheaper costs and high mechanical strength, the diffusion of small gas molecules such as oxygen through PET films remains a matter of great concern. The simple yet robust supramolecular deposition of GNP/polyelectrolyte on PET substrates significantly increases the tortuous path the oxygen molecule has to travel, making it harder to diffuse through the PET film. With permeability coefficients in the range of 10-18 cc cm/cm(2) s Pa, the coatings developed in this study show three orders of magnitude reduction as compared to the permeability coefficient of the bare PET film, significantly lower than that of ethylene vinyl alcohol (EVOH) and comparable to silicon oxide thin films used in commercial gas barrier foils. The use of GNPs in the multilayered films also helped reduce the electrical sheet resistance to about 1MΩ which is five orders of magnitude lower than the original PET substrate opening up promising opportunities for future use in semiconductor and electronics industry. Making suitable modifications in the deposition process, three configurations of GNP embedded PEM multilayers namely hydrogen bonded, electrostatic, and composite films were developed and their effect on oxygen barrier property and sheet resistance was monitored. Oxygen permeability of films was tested in accordance with ASTM D-3985 using a MOCON 2/21 ML instrument, whereas electrical sheet resistance was quantified using a Gamry Femtostat Electrochemical Impedance station.


Aphasic seizures in patients with temporopolar and anterior temporobasal lesions: a video-EEG study.
Studies of patients with temporal lobe epilepsy provide few descriptions of seizures that arise in the temporopolar and the anterior temporobasal brain region. Based on connectivity, it might be assumed that the semiology of these seizures is similar to that of medial temporal lobe epilepsy. However, accumulating evidence suggests that the anterior temporobasal cortex may play an important role in the language system, which could account for particular features of seizures arising here. We studied the electroclinical features of seizures in patients with circumscribed temporopolar and temporobasal lesions in order to identify specific features that might differentiate them from seizures that originate in other temporal areas. Among 172 patients with temporal lobe seizures registered in our epilepsy unit in the last 15 years, 15 (8.7%) patients had seizures caused by temporopolar or anterior temporobasal lesions (11 left-sided lesions). The main finding in our study is that patients with left-sided lesions had aphasia during their seizures as the most prominent feature. In addition, while all patients showed normal to high intellectual functioning in standard neuropsychological testing, semantic impairment was found in a subset of 9 patients with left-sided lesions. This case series demonstrates that aphasic seizures without impairment of consciousness can result from small, circumscribed left anterior temporobasal and temporopolar lesions. Thus, the presence of speech manifestation during seizures should prompt detailed assessment of the structural integrity of the basal surface of the temporal lobe in addition to the evaluation of primary language areas.


The effect of tibolone versus 17beta-estradiol on climacteric symptoms in women with surgical menopause: a randomized, cross-over study.
To compare the effectiveness of tibolone and 17beta-estradiol on climacteric symptoms, in a randomized, single-blind, cross-over study in surgically menopausal women. Forty surgically menopausal women were divided randomly into two groups. Group A received treatment with tibolone for 6 months, while group B received 17beta-estradiol. After 3 weeks washout period, treatment protocols were exchanged for another 6 months. The climacteric symptoms were assessed with Greene Climacteric Scale at baseline, during washout and after the treatments. Statistical analysis was done with the Wilcoxon's Sign Rank test. Both treatments significantly improved the scores of all subscales with respect to baseline. However, the improvement in psychological, somatic and sexual subscales were significantly superior in the tibolone group compared with 17beta-estradiol group. Both treatments showed comparable improvements in the relief of vasomotor symptoms. Our findings suggest that tibolone may improve mood, libido and somatic symptoms in surgically menopausal women to a greater extent than estrogen therapy alone.