Analysis of non-linearities in mutation frequency curves.
Mutation frequency curves for ultraviolet light and other mutagens often exhibit non-linear, as well as linear components. A common pattern observed for UV-induced reversion of auxotrophy in yeast is a biphasic, linear-quadratic (or higher order) response. The non-linear component in such a biphasic frequency curve can arise in two distinct, but non-mutually exclusive, ways: (i) as a result of the existence of two-hit processes in the molecular mechanism(s) of mutagenesis; and (ii) as a result of the possible stochastic dependence of mutation and killing, such that the probability of clone formation by the mutant cells differs from that of the non-mutant cells in the population. We describe here a simple mathematical method for distinguishing between these two sources of non-linearity. It is based on the calculation of a quantity that we call 'apparent survival.' This is given, for any mutagen dose chi, by the ratio of the mutant yield to the corresponding linear component of mutation frequency. If the apparent survival rises to values greater than unity before declining at high doses, then there must exist positive two-hit (or higher order) components in the mutational mechanism. If the final slope of the apparent survival curve differs from that of the measured survival curve, then there also exists some degree of stochastic dependence between mutation and killing.


The murine gene for cellular retinoic acid-binding protein type II. Genomic organization, chromosomal localization, and post-transcriptional regulation by retinoic acid.
The cellular retinoic acid-binding protein type II (CRABP-II) is a member of the serum and cytoplasmic retinoid-binding protein family. It is expressed during embryonic development and in adult skin and is upregulated by retinoic acid (RA) in F9 teratocarcinoma cells. We have determined the genomic organization of the murine CRABP-II gene and performed a detailed analysis of its transcriptional unit. The CRABP-II gene, located on mouse chromosome 2, is approximately 4.6 kilobases long and divided into four exons in a structure common to other members of the family of serum and cellular retinoid-binding proteins. Primer extension analysis and S1 nuclease protection assay were used to identify the transcription initiation site which is located 27 base pairs downstream of a typical TATAA box. Sequence analysis of the promoter also revealed a GC-rich region with overlapping putative SP1-binding sites at nucleotides -61 and AP-1 and AP-2-binding sites at nucleotides -518 and -544, respectively. The 3'-untranslated region contains two copies of the pentanucleotide AUUUA shown to be involved in messenger RNA destabilization. Consensus sequence for retinoic acid response elements were not detected in the promoter region of the CRABP-II gene. Results of nuclear run on experiments show that the CRABP-II gene is not transcriptionally activated by RA in F9 teratocarcinoma cells. These results suggest that the up-regulation of CRABP-II mRNA levels by RA is mainly controlled by a post-transcriptional mechanism.


Severe fatigue and related factors in cancer patients before the initiation of treatment.
It is generally known that fatigue is a common symptom during cancer treatment, and in cancer survivors. However, fatigue was never studied after diagnosis, before cancer treatment was initiated. This study investigated the prevalence of severe fatigue, and related factors, in cancer patients before the initiation of treatment. One hundred and seventy-nine patients with various malignancies were assessed before start of treatment with curative intention, including the Checklist Individual Strength, Sickness Impact Profile, Beck Depression Inventory Primary Care, Symptom Checklist-90, and six Numeric Rating Scales to measure fatigue, pain and physical activity. To test which factors contributed to severe fatigue a logistic regression analysis was performed. In total 23.5% patients were severely fatigued, varying between diagnoses; prostate cancer (14.3%), breast cancer (20.3%), and gastrointestinal cancer (28.1%). Currently lower physical activity (P=0.013), more depressive mood (P=0.014), impaired sleep and rest during the day and night (P=0.045), and fatigue 1 year before diagnosis (P=0.005) contributed to severe fatigue. Relatively large numbers of cancer patients already experience severe fatigue before initiation of treatment, varying between 14-28%. The factors that contributed to severe fatigue at this stage were physical activity, depressive mood, impaired sleep and rest, and fatigue 1 year before diagnosis.


Status of the glass sponge reefs in the Georgia Basin.
The purpose of this paper is to describe the status and general faunal composition of sponge reefs in the Georgia Basin (GB), British Columbia, Canada. Fourteen distinct deep-water glass sponge (Hexactinellid) reefs have been mapped using multibeam bathymetry and sidescan sonar in the GB. Seven of these have been surveyed visually using video from remotely operated vehicles (ROVs). Analysis of video data indicated that three reefs were undamaged, two were damaged and the other two were damaged but potentially recovering. The nature of the damaged reefs, with large areas of scattered dead sponge skeleton fragments and few live reef-building sponges (Aphrocallistes vastus and Heterochone calyx), as well as video evidence of tracks suggest they were damaged mechanically by mobile fishing gear. Relative abundance of the megafauna associated with the reefs is discussed in the context of oceanographic conditions, such as sediment accumulation and organic flux, as well as overall reef status. Of particular interest for fisheries conservation efforts in the area was the fact that one undamaged reef in the southern GB showed higher taxonomic richness and abundance of rockfish (Sebastes spp.), both adult and juvenile, compared to an adjacent damaged reef. This result suggests that undamaged reefs may act as refugia for these endangered stocks.


Influence of the molecular structure of N6-(omega-aminoalkyl)adenosines on adenosine receptor affinity and intrinsic activity.
The affinities of a series of N6-(omega-aminoalkyl)adenosines as probes for A1 and A2 adenosine receptors were determined in various radioligand binding assays and the intrinsic activities were measured in adenylate cyclase assays. Clear species differences were noticed for A1 receptor affinity of these adenosine receptor agonists, the compounds being more active in calf than in rat brain tissue. The affinity profile within the series was, however, rather similar in both membrane preparations, with N6-9-aminononyladenosine displaying highest affinity. The A2 receptor affinities were comparable to values measured for the A1 receptor in its low affinity state, as assessed with a radiolabelled antagonist in the presence of 1 mM GTP. Calculation of the intrinsic activities of the adenosine analogues from their modulating action on adenylate cyclase showed almost all the compounds to be equally effective to (-)-N6-(R-phenylisopropyl) adenosine, on either A1 or A2 adenosine receptors. N6-3-Aminopropyl- and N6-12-aminododecyladenosine, however, proved to be partial agonists, the first on A1 and the second on A2 adenosine receptors. The data are used as the basis for a discussion of adenosine receptor subtype selectivity and intrinsic activity in general.


The T-type calcium channel antagonist Z944 rescues impairments in crossmodal and visual recognition memory in Genetic Absence Epilepsy Rats from Strasbourg.
Childhood absence epilepsy (CAE) is often comorbid with behavioral and cognitive symptoms, including impaired visual memory. Genetic Absence Epilepsy Rats from Strasbourg (GAERS) is an animal model closely resembling CAE; however, cognition in GAERS is poorly understood. Crossmodal object recognition (CMOR) is a recently developed memory task that examines not only purely visual and tactile memory, but also requires rodents to integrate sensory information about objects gained from tactile exploration to enable visual recognition. Both the visual and crossmodal variations of the CMOR task rely on the perirhinal cortex, an area with dense expression of T-type calcium channels. GAERS express a gain-in-function missense mutation in the Cav3.2 T-type calcium channel gene. Therefore, we tested whether the T-type calcium channel blocker Z944 dose dependently (1, 3, 10mg/kg; i.p.) altered CMOR memory in GAERS compared to the non-epileptic control (NEC) strain. GAERS demonstrated recognition memory deficits in the visual and crossmodal variations of the CMOR task that were reversed by the highest dose of Z944. Electroencephalogram recordings determined that deficits in CMOR memory in GAERS were not the result of seizures during task performance. In contrast, NEC showed a decrease in CMOR memory following Z944 treatment. These findings suggest that T-type calcium channels mediate CMOR in both the GAERS and NEC strains. Future research into the therapeutic potential of T-type calcium channel regulation may be particularly fruitful for the treatment of CAE and other disorders characterized by visual memory deficits.


Morphine upregulates Toll-like receptor 4 expression and promotes melanomas in mice.
Morphine and other opioids are used to manage cancer-related pain; however, the role of these drugs in cancer progression remains controversial. Emerging evidence indicates that morphine can activate Toll-like receptor 4 (TLR4) and its signaling pathways, by the way the activation and expression of TLR4 can promote melanoma. In this study, we investigated the effects of morphine on the expression of TLR4 and promotion of melanoma in mice. Mice melanoma cells (B16F10) were cultured with morphine (0.1, 1 and 10 µM) for 24 h. In the other experiment, cells were treated with morphine with or without TLR4 agonist (LPS) or antagonist (TAK-242). In in-vivo model, B16F10 cells were subcutaneously injected to C57BL/6 mice, and morphine was administrated in three different treatment protocols after developing palpable tumors (acute treatment, chronic daily injections, escalating doses of morphine). In another set of experiments, B16F10 cells were pretreated with LPS (5 µg/ml) 24 h before injection into mice. Control group received normal saline. We measured cell proliferation, the expression level of Tlr4, Nuclear factor kappa-light-chain-enhancer of activated B cells 1 (Nf-κb1) genes, TLR4 protein expression, and tumor volume. Chronic, acute, and escalating doses of morphine increased tumor. Morphine increased the expression of Tlr4 and Nf-κb1 regardless of the treatment protocol used. Morphine increases the progression of melanoma cancer and may be related to the increased expression of TLR4. Our results suggest that morphine should be used with caution in patients with melanoma.HighlightsMorphine increases the expression of TLR4 in melanoma.Morphine increases melanoma progression.These effects are mostly observed with chronic and escalating morphine administration.


Does O-methyl-dopa play a role in levodopa-induced dyskinesias?
Clinically scored levodopa-induced dyskinesias were correlated with plasma dopa and O-methyl-dopa levels determined every hour during one day in 30 Parkinsonian-treated patients. In patients treated with a combination of L-dopa and a peripheral decarboxylase inhibitor (PDI), those with dyskinesias have very high plasma O-methyl-dopa levels compared with those who have no dyskinesias. In contrast, no significantly different plasma dopa levels are found in these two subgroups of patients, leaving open the question of the possible involvement of such elevated plasma O-methyl-dopa levels in favouring dyskinesias.


Structure modeling from small angle X-ray scattering data with elastic network normal mode analysis.
Computational algorithms to construct structural models from SAXS experimental data are reviewed. SAXS data provides a wealth of information to study the structure and dynamics of biological molecules, however it does not provide atomic details of structures. Thus combining the low-resolution data with already known X-ray structure is a common approach to study conformational transitions of biological molecules. This review provides a survey of SAXS modeling approaches. In addition, we will discuss theoretical backgrounds and performance of our approach, in which elastic network normal mode analysis is used to predict reasonable conformational transitions from known X-ray structures, and find alternative conformations that are consistent with SAXS data.


CMAPs in pharyngeal and hyoid muscles evoked by nucleus solitarius stimulation in dogs.
Electrical stimulation of the solitary nucleus with a single pulse of 150 to 500 microA caused visible contractions of the muscles involved in swallowing but did not activate the central pattern generator for swallowing. Electromyographically there were short bursts of activity characterized by an onset of activity within 1 to 7 ms, a duration of 14 to 26 ms, and an amplitude of 10 to 4,800 microV. The CMAPs were recorded in the pharyngeal and hyoid muscles on both the ipsilateral and the contralateral sides of the stimulated solitary nucleus. Across all five pairs of muscles in which the recordings were made the latencies of the CMAPs in the contralateral muscles were significantly greater and the amplitudes significantly lower, but the durations were not significantly different. Bilateral activation of the muscles by unilateral stimulation may not result in symmetrical contraction of the pharyngeal muscles.


Elucidation of the antigenic determinant in lipotropin for two monoclonal antibodies.
Two monoclonal antibodies were found to give enhanced affinity for beta-lipotropin when mixed, as evidenced by competitive radioimmunoassay. Both monoclonals were found to react with a pentapeptide Ala-Glu-Leu-Glu-Tyr, which is a sequence of high local hydrophilicity within the N-terminal section of beta-lipotropin.


Ondansetron in the prevention of postoperative nausea and vomiting after laparoscopic cholecystectomy: a prospective randomized study.
There is an increased incidence of postoperative nausea and vomiting (PONV) after laparoscopic cholecystectomy. The aim of the study was to evaluate the efficacy of intravenous ondansetron, a 5-HT anatagonist, for prevention from PONV after laparoscopic cholecystectomy. Sixty-eight patients were entered into the study. Thirty-six patients were randomized to receive a single intravenous dose of 4 mg of ondansetron before extubation. Thirty-two patients received no prophylaxis. There were no differences in terms of sex ratio, age, ASA status, and duration of operation between the two study groups. Patients were interviewed by an independent observer to assess the postoperative pain and nausea using visual analog score. Patients' satisfaction scores to the procedure were measured. There was no complication in the series. Pain scores at 2 and 24 h after operation and analgesics consumption were not different between the two groups. For the patients who received ondansetron, the mean (SD) nausea score at 2 and 24 h was 1.5 (1.2) and 1.4 (1.4), respectively. The score for the control group was 1.4 (1.0) and 1.2 (0.5), respectively. There was also no difference in episodes of vomiting and usage of antiemetics between the two groups. Both groups were equally satisfied with the procedures. Routine use of ondansetron does not reduce the incidence of postoperative nausea and vomiting after laparoscopic cholecystectomy.


The daily goals communication sheet: a simple and novel tool for improved communication and care.
This tool directs the health care team to discuss specific goals of care and best-practice safety measures daily for each patient and to ensure goal understanding and implementation.


Oxygen-17 compounds as potential NMR T2 contrast agents: enrichment effects of H2(17)O on protein solutions and living tissues.
The isotopic enrichment of solutions, living tissues, and organisms with oxygen-17 in the form of H2(17)O shortens their proton NMR transverse relaxation times (T2) and produces changes in NMR image intensity. The transverse relaxation rate (1/T2) was found to be linearly dependent on the H2(17)O concentration in biological solutions up to 5% enrichment. The longitudinal relaxation time (T1) is not affected by enrichment. Equal concentrations of H2(17)O do not produce the same magnitude of T2 change in all physiological environments. The reasons for these differences are discussed. The results suggest that certain oxygen-17 compounds should be explored as "contrast agents" in magnetic resonance imaging.


Solar radiation and age-related macular degeneration.
Age-related macular degeneration (AMD) involves a progressive impairment of the outer layers in the center of the retina. Experimental studies have demonstrated that bright light preferentially damages precisely the region that degenerates in AMD. The evidence that solar radiation is responsible for some of the deteriorative changes that lead to AMD is examined in this review. In the primate eye, the high-energy portion of the solar spectrum is most hazardous to retinal molecules, with damaging effects increasing as photon energy rises. This action spectrum is explicable by the quantum laws which describe the interaction of radiation with matter. High-energy visible and ultraviolet photons can produce molecular damage by a photochemical mechanism. The lesion is exacerbated by oxygen, which initiates free-radical chain reactions (photodynamic effects). Melanin exerts a protective effect against damage from sunlight. In the human retina, documented lesions from solar radiation range from the acute effects of sun-gazing to injuries resulting from prolonged periods of exposure in brightly illuminated environments. The damage occurs in the same region that degenerates in AMD. A cataractous lens and ocular melanin both protect the retina against AMD, as predicted by the radiation hypothesis. Identification of an environmental factor that evidently plays a role in the etiology of AMD provides the basis for a program of preventive medicine.


Effects of haloperidol, lithium, and valproate on phosphoinositide turnover in rat brain.
The effects of acute, subacute, and chronic treatment with haloperidol, lithium, and valproate on inositol phosphate (IP) formation were examined. Acute treatment with haloperidol or the combination of haloperidol and lithium significantly reduced IP basal cortical levels. Subacute (three days) treatment with lithium decreased the IP basal level in the frontal cortex. Chronic treatment with haloperidol (14 and 28 days) caused a significant attenuation of carbachol-sensitive IP accumulation in the frontal cortex and striatum and a significant decrease in norepinephrine (NE)-induced IP formation in the frontal cortex (14 and 28 days) and striatum (28 days). Lithium treatment for 14 days produced a significant reduction in the IP basal cortical value, and a significant reduction in cortical carbachol- and NE-induced IP formation was found after 28 days of lithium treatment. The combination of haloperidol and lithium for 28 days decreased the striatal carbachol- and cortical NE-induced IP accumulation and caused a significant increase in NE-sensitive IP formation in the striatum at 14 days. Valproate treatment for 28 days was associated with a significant attenuation in striatal agonist-stimulated IP formation. Therefore, three drugs with different specificities for primary neurotransmitters may have common effects on second-messenger systems.


Regional distribution of guanine nucleotide-sensitive and guanine nucleotide-insensitive vasoactive intestinal peptide receptors in rat brain.
Based on the ability of guanine nucleotides to inhibit the binding of vasoactive intestinal peptide to its receptors, a guanosine 5'-triphosphate analog, guanylyl-imidodiphosphate, was used to differentiate two subtypes (or different functional states of a single subtype) of vasoactive intestinal peptide receptor in brain with in vitro autoradiography. In most brain regions, guanylyl-imidodiphosphate reduced vasoactive intestinal peptide binding between 40 and 60%. However, in the supraoptic nucleus, locus coeruleus, interpeduncular nucleus, facial nucleus, olfactory tubercle and periventricular hypothalamic nucleus, 80% or more of vasoactive intestinal peptide binding was inhibited. In other brain regions, including the medial geniculate, olfactory bulbs, and ventral thalamic nuclei, guanylyl-imidodiphosphate had little effect on vasoactive intestinal peptide binding. In liver, lung and intestine it also partly inhibited vasoactive intestinal peptide binding. Electrophoretic analysis of vasoactive intestinal peptide, covalently cross-linked to its receptors in brain membranes, revealed a pair of bands between 44,000 and 52,000 mol. wt, a component at 64,000 mol. wt and another at 92,000 mol. wt. All were displaceable with vasoactive intestinal peptide but guanylyl-imidodiphosphate displaced only the 64,000 mol. wt band suggesting that the GTP-sensitive vasoactive intestinal peptide receptor seen in brain sections has a molecular weight of about 61,000. The differential sensitivity to guanylyl-imidodiphosphate suggests the existence of at least two vasoactive intestinal peptide receptor subtypes in brain, with distinct regional distribution, and may reflect differential coupling to second messenger systems.


Magnetic resonance radiomics-derived sphericity correlates with seizure in brain arteriovenous malformations.
Angioarchitectural analysis of brain arteriovenous malformations (BAVMs) is qualitative and subject to interpretation. This study quantified the morphology of and signal changes in the nidal and perinidal areas by using MR radiomics and compared the performance of MR radiomics and angioarchitectural analysis in detecting epileptic BAVMs. From 2010 to 2020, a total of 111 patients with supratentorial BAVMs were retrospectively included and grouped in accordance with the initial presentation of seizure. Patients' angiograms and MR imaging results were analyzed to determine the corresponding angioarchitecture. The BAVM nidus was contoured on time-of-flight MR angiography images. The perinidal brain parenchyma was contoured on T2-weighted images, followed by radiomic analysis. Logistic regression analysis was performed to determine the independent risk factors for seizure. ROC curve analysis, decision curve analysis (DCA), and calibration curve were performed to compare the performance of angioarchitecture-based and radiomics-based models in diagnosing epileptic BAVMs. In multivariate analyses, low sphericity (OR: 2012.07, p = .04) and angiogenesis (OR: 5.30, p = .01) were independently associated with a high risk of seizure after adjustment for age, sex, temporal location, and nidal volume. The AUC for the angioarchitecture-based, MR radiomics-based, and combined models was 0.672, 0.817, and 0.794, respectively. DCA confirmed the clinical utility of the MR radiomics-based and combined models. Low nidal sphericity and angiogenesis were associated with high seizure risk in patients with BAVMs. MR radiomics-derived tools may be used for noninvasive and objective measurement for evaluating the risk of seizure due to BAVM. Low nidal sphericity was associated with high seizure risk in patients with brain arteriovenous malformation and MR radiomics may be used as a noninvasive and objective measurement method for evaluating seizure risk in patients with brain arteriovenous malformation. • Low nidal sphericity was associated with high seizure risk in patients with brain arteriovenous malformation. • The performance of MR radiomics in detecting epileptic brain arteriovenous malformations was more satisfactory than that of angioarchitectural analysis. • MR radiomics may be used as a noninvasive and objective measurement method for evaluating seizure risk in patients with brain arteriovenous malformation.


Cytophotometric DNA analysis of early esophageal carcinoma.
In four patients with early esophageal carcinoma who underwent radical surgery, one died of cancer recurrence 13 months after surgery, two had an uneventful postoperative course after over five years and one succumbed to an incidental pneumonia 18 months postoperatively. Cytophotometric DNA analysis of the cancer cells reflected well the outcome of these patients. One recurrent patient had malignant pattern similar to that seen in usual advanced carcinoma of the esophagus, whereas the remaining three patient without recurrence throughout the postoperative course showed less malignant patterns. These result suggested the potential usefulness of cytophotometric DNA analysis in assessing the prognosis of early esophageal carcinoma.


A Masking Effect: A Case of Initial Presentation of Ulcerative Colitis After Discontinuing Growth Hormone Therapy.
The inflammation and repair of the intestinal mucosa in inflammatory bowel disease (IBD) involve a complex interplay between innate, adaptive immune responses, and hormones. This may explain the relapsing clinical course of the disease. We present the first reported case of a patient presenting their initial flare of ulcerative colitis immediately after discontinuing growth hormone (GH) therapy, suggesting treatment with GH or growth factors may prevent the development of IBD. This is a case of a 13-year-old female with a history of GH deficiency, presenting with an 8-week history of abdominal pain, blood-stained diarrhea, and fecal calprotectin greater than 8000 mcg/g, 2 weeks after discontinuing GH therapy. The patient subsequently underwent an esophagoduodenoscopy and colonoscopy with biopsies showing histological features consistent with ulcerative colitis. The finding of withdrawing GH or growth factors therapy potentially unmasking IBD in this patient raises a question of whether growth factors can inhibit the development of IBD and suggests beneficial effects of treatment with GH or growth factors as adjuvant therapy for IBD.


Multimodality imaging of benign and malignant male breast disease.
With the increasing use of advanced imaging techniques, male breast lesions are being visualized using techniques other than mammography and ultrasound. This review illustrates benign and malignant male breast disease on both conventional imaging as well as advanced imaging methods including computed tomography, magnetic resonance imaging, and positron-emission tomography in order to familiarize the radiologist with typical imaging appearances and review the proper clinical management.


Activation of protein kinase R in the manganese-induced apoptosis of PC12 cells.
Manganese neurotoxicity leads to Parkinson-like symptoms associated with the apoptotic cell death of dopaminergic neurons. Protein kinase R (PKR) is a serine/threonine-specific protein kinase that has been implicated in several cellular signal transduction pathways, including the induction of apoptosis. Here, we investigated the role of PKR in the manganese-induced apoptosis of dopamine-producing pheochromocytoma PC12 cells. Manganese (0.5 mM) induced the proteolytic cleavage of PKR and caspase-3, DNA fragmentation, and cell death, which were prevented by the co-treatment of PC12 cells with a PKR specific inhibitor, C16 in a concentration-dependent manner. C16 did not affect the manganese-induced activation of the c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (MAPK) pathway, indicating that PKR functions downstream of JNK and p38 MAPK. In contrast, C16 triggered the activation of the p44/42 MAPK (ERK1/2) pathway and induced hemoxygenase-1, both in the absence and presence of manganese. PKR is reportedly involved in endoplasmic reticulum (ER) stress-induced apoptosis. Manganese activated all three branches of the unfolded protein response in PC12 cells; however, this effect was very weak compared with the ER stress induced by the well-known ER stress inducers thapsigargin and tunicamycin. Moreover, C16 did not affect manganese-induced ER stress at concentrations that almost prevented caspase-3 activation and DNA fragmentation. These results suggest that PKR is involved in manganese-induced apoptotic cell death and stress response, such as the activation of the p44/42 MAPK pathway and the induction of hemoxygenase-1. Although manganese induced a faint, but typical, ER stress, these events contributed little to manganese-induced apoptosis.


Racial contrasts of blood pressure levels in two southern, rural communities.
Blood pressure and anthropometric measurements were obtained according to a standardized protocol in two biracial, rural, southern communities. Essentially, the same examination team and protocol were used in Franklinton and Bogalusa. No differences between the communities in mean height or weight (except for white boys) were observed. Blood pressure levels were 2-5 mm Hg higher in Franklinton, particularly in blacks ages 8-13 years. These differences could not be explained by differences in anthropometric measurements or observers. The difference may be due partly to the fact that larger numbers of students each morning were examined in Franklinton than Bogalusa in perhaps a more hurried environment. Most of the staff members, being from Bogalusa, were not familiar to the children in Franklinton (as they are in Bogalusa), and this may have created anxiety. The differences were small, however, when compared with differences observed among several populations using varying methods. Confirmation is given to the finding that height and weight, not age, are the strongest determinants of blood pressure level in growing children, giving a further clue toward development of an appropriate definition of hypertension in children.


Torsades de pointes as a cause of sudden death in a patient with aortic stenosis and atrial fibrillation.
The occurrence of sudden cardiac death during Holter monitoring in patients with aortic stenosis has been reported previously. In the majority of the reported cases, the cause of death was a malignant ventricular tachyarrhythmia. The presence of a strong association between frequency and complexity of ventricular arrhythmias and sudden death in patients with aortic stenosis has been proposed. We report the case of a 77-year-old woman with aortic stenosis and atrial fibrillation who had an episode of torsades de pointes that degenerated into ventricular fibrillation during Holter monitoring. A short-long-short sequence, but not increased ventricular ectopics, precipitated torsades de pointes and sudden death in this case which is strongly indicative of triggered activity as the underlying mechanism of the lethal arrhythmia.


Aggregates of particles on the plasmalemma of striated muscle from a spider.
Aggregates of large intramembranous particles are described in the surface membrane of muscle fibers of a spider. The particles are continuous with extensions into the extracellular space, forming large complexes. No intramembranous particles are visible at sites of Z line attachment to the plasma membrane.


Ex vivo transduction of human dermal tissue structures for autologous implantation production and delivery of therapeutic proteins.
Systemic delivery of therapeutic proteins through gene transfer approaches has been carried out mostly by ex vivo transduction of single cells or by direct in vivo injection of an expression vector. In this work an intact miniature biopsy of human dermis (microdermis) is harvested and transduced ex vivo by a viral vector encoding a gene for the therapeutic protein. The microdermis preserves its three-dimensional structure and viability during the ex vivo manipulations. Furthermore, upon transduction with adenoviral and adeno-associated viral vectors the microdermis secretes recombinant human erythropoietin (hEPO). Biochemical analysis of the secreted hEPO showed similarity to the clinically approved recombinant hEPO. Subcutaneous implantation of microdermal hEPO into SCID mice exhibited hEPO secretion in the blood circulation and preserved elevated hematocrit for several months, demonstrating the technology's potential for sustained delivery of protein therapeutics.


Utilization of a kinetic isotope effect to decrease decomposition of ceftriaxone in a mixture of D2O/H2O.
The discovery of cephalosporin and demonstration of its improved stability in aqueous solution, as well as enhanced in vitro activity against penicillin-resistant organisms, were major breakthroughs in the development of β-lactam antibiotics. Although cephalosporins are more stable with respect to hydrolytic degradation than penicillins, they still experience a variety of chemical transformations. The present study offers an insight into the rates and mechanisms of ceftriaxone degradation at the therapeutic concentration in water, a mixture of water and deuterium oxide, and deuterium oxide itself at the neutral pH. Specific ceftriaxone degradation products were observed in aged samples (including a previously unreported dimer-type species), and by comparing the degradation rates in H2O and D2O, the observation of a kinetic isotope effect provided some valuable insight as to the nature of the initial ceftriaxone degradation. The effect of protium to deuterium isotope change on the degradation kinetics of ceftriaxone was evaluated using the method of initial rates based on HPLC analysis as well as by quantitative 1H NMR spectroscopy. Moreover, computational analysis was utilized to get a molecular insight into chemical processes governing the ceftriaxone degradation and to rationalize the stabilizing effect of replacing H2O with D2O.


Developments in protein microsequencing.
The author has described here several methods which have been developed in his laboratory. These methods are explained in the following order: [I] gel electrophoresis; 1) extraction from the conventional polyacrylamide gel, 2) a new polyacrylamide gel electrophoresis system for small peptides. [II] Amino acid composition (rapid and micro methods for hydrolysis of proteins); 1) hydrolysis with a mixture of trifluoroacetic acid and hydrochloric acid, 2) vapour hydrolysis, 3) tryptophan micro-analysis. [III] Amino-terminal sequencing; 1) a sensitive detection method of Edman degradation, 2) N-terminal specific labeling method. [IV] Carboxy-terminal sequencing by carboxypeptidase digestion; 1) field desorption mass spectrometry, 2) the use of detergents or alcohols. [V] Monitoring of amino groups at the DNA binding site of protein. As mentioned in the beginning, techniques in the protein field tend to be behind comparable techniques in the DNA field. In addition, the fact that the most important biological molecules presently studied are made in such small amounts in the cell has demanded the development of techniques using precisely and quickly only micro amounts of protein. Such developments of protein chemistry techniques for purification, composition, sequencing, and chemical modification are very much needed in the cell biology field and close collaboration is required between protein chemistry, biophysics, and modern biology in order to develop the above methodology.


The composition, seasonal variation, and potential sources of the atmospheric wet sulfur (S) and nitrogen (N) deposition in the southwest of China.
The composition, seasonal variation, and potential sources of sulfate (S) and nitrogen (N) deposition in precipitation in the southwest of China from 2003 to 2013 were investigated. The results showed that the concentration of SO4 (2-), NO3 (-), and NH4 (+) in rainwater were 10.57-1360, 7.16-523.71, and 7.54-1020 μeq l(-1), with an annual volume-weighted mean (VWM) concentration of 103.99, 46.73, and 97.30 μeq l(-1), respectively. The annual wet deposition of SO4 (2-), NO3 (-), and NH4 (+) was 21.66, 8.16, and 17.49 kg S (N) ha(-1), respectively. The temporal variations of the ions showed that the abrupt decreasing breakpoints were in 2008 for SO4 (2-) and in 2009 for NO3 (-) and NH4 (+), and increasing trends were observed after 2010 for the three ions. These trends reflected the effect of economy recession and the policy of controlling SO2 and NOx emissions. The acid rain type of precipitation was shifted from sulfur to a mixed one. The ions of SO4 (2-), NO3 (-), and NH4 (+) presented high values in winter and spring and low values in autumn and summer. A highly positive linear correlation between SO4 (2-) and NO3 (-) (R(2) = 0.71), SO4 (2-) and NH4 (+) (R(2) = 0.74), and NO3 (-) and NH4 (+) (R(2) = 0.84) existed while a strong negative correlation was found between the three main ionic concentrations and precipitation. The SO4 (2-) was mainly from fossil fuel combustion (60.53%), aged sea salt (19.03%), agriculture (11.38%), crust (6.66%), and biomass burning (2.40%); the NO3 (-) was mainly from fossil fuel combustion (75.41%), biomass burning (9.67%), aged sea salt (7.97%), and agriculture (6.96%); and the NH4 (+) was mainly from agriculture (86.38%), fossil fuel combustion (10.52%), and aged sea salt (3.09%).


Long-Term Outcomes in Complex Abdominal Wall Reconstruction Repaired With Absorbable Biologic Polymer Scaffold (Poly-4-Hydroxybutyrate).
After promising early outcomes in the use of absorbable biologic mesh for complex abdominal wall reconstruction, significant criticism has been raised over the longevity of these repairs after its 2-year resorption profile. This is the long-term (5-year) follow-up analysis of our initial experience with the absorbable polymer scaffold poly-4-hydroxybutyrate (P4HB) mesh compared with a consecutive contiguous group treated with porcine cadaveric mesh for complex abdominal wall reconstructions. Our clinical analysis was performed using Stata 14.2 and Excel 16.16.23. After a 5-year follow-up period, the P4HB group (n = 31) experienced lower rates of reherniation (12.9% vs 38.1%; P = 0.017) compared with the porcine cadaveric mesh group (n = 42). The median interval in months to recurrent herniation was similar between groups (24.3 vs 20.8; P = 0.700). Multivariate logistic regression analysis on long-term outcomes identified smoking (P = 0.004), African American race (P = 0.004), and the use of cadaveric grafts (P = 0.003) as risks for complication while smoking (P = 0.034) and the use of cadaveric grafts (P = 0.014) were identified as risks for recurrence. The long-term cost analysis showed that P4HB had a $10,595 per case costs savings over porcine cadaveric mesh. Our study identified the superior outcomes in clinical performance and a value-based benefit of absorbable biologic P4HB scaffold persisted after the 2-year resorption timeframe. Data analysis also confirmed the use of porcine cadaveric grafts independently contributed to the incidence of complications and recurrences.


Modulating the physicochemical and biological properties of carbon dots synthesised from plastic waste for effective sensing of E. coli.
The reliable means for the quick recognition of pathogenic bacteria for disease prevention has acquired a considerable significance among researchers for human wellbeing. The purpose of current study is to fabricate highly photo-luminescent and biocompatible carbon dots (CQDs) from single used plastic waste such as poly bags, cups and bottles for selective sensing of E. coli. The formed small sized spherical CQDs have possessed high water solubility with excellent photo stable nature. The detailed fluorescence emission studies of CQDs have illustrated the overview of different types of surface defects and emissive states in formed particles. In addition, the specific role of prepared CQDs with bio-compatible nature has been thoroughly checked on hemolysis of red blood cells by employing in-vitro blood compatibility assay. A combination of anticoagulant and thrombolytic activity of CQDs with anti-oxidative activities from reactive oxygen species (ROS) generation were found to be responsible for the biocompatibility of formed CQDs. The effective in vitro stability toward histidine, cysteine, bovine serum albumin (BSA), human serum albumin (HSA) and saline supported the biocompatibility of formed particles as a function of different concentrations of CQDs. The optimized fluorescence properties with high surface to volume ratio of CQDs have provided a better alternative for "turn-off" fluorescence detection of E. coil in aqueous media with limit of detection down to 108 CFU/mL estimated by using standard colony-counting method. The as prepared nanosensor provided easy fabrication with fast response, high sensitivity and selectivity in real water samples which might be necessary for detecting pathogenic agents. In addition, the current work has provided the large overview in blood compatibility and anti-oxidative potential of prepared CQDs and giving useful information for preparing blood-compatible nanoparticles with enhanced bacterial discrimination ability.


Unsuccessful treatment outcome and associated risk factors. A prospective study of DR-TB patients from a high burden country, Pakistan.
Tuberculosis (TB), a curable and preventable infectious disease, becomes difficult to treat if resistance against most effective and tolerable first line anti-TB drugs is developed. The objective of the present study was to evaluate the treatment outcomes and predictors of poor outcomes among drug-resistant tuberculosis (DR-TB) patients treated at a programmatic management unit of drug resistant tuberculosis (PMDT) unit, Punjab, Pakistan. This prospective observational study was conducted at a a PMDT unit in Multan, Punjab, Pakistan. A total of 271 eligible culture positive DR-TB patients enrolled for treatment at the study site between January 2016 and May 2017 were followed till their treatment outcomes were recorded. World Health Organization's (WHO) defined criteria was used for categorizing treatment outcomes. The outcomes of cured and treatment completed were collectively placed as successful outcomes, while death, lost to follow-up (LTFU) and treatment failure were grouped as unsuccessful outcomes. Multivariable binary logistic regression analysis was employed for getting predictors of unsuccessful treatment outcomes. A p-value <0.05 was considered statistically significant. Of the 271 DR-TB patients analysed, nearly half (51.3%) were males. The patient's (Mean ± SD) age was 36.75 ± 15.69 years. A total of 69% patients achieved successful outcomes with 185 (68.2%) patients being cured and 2 (0.7%) completed therapy. Of the remaining 84 patients with unsuccessful outcomes, 48 (17.7%) died, 2 (0.7%) were declared treatment failure, 34 (12.5%) were loss to follow up. After adjusting for confounders, patients' age > 50 years (OR 2.149 (1.005-4.592) with p-value 0.048 and baseline lung cavitation (OR 7.798 (3.82-15.919) with p-value <0.001 were significantly associated with unsuccessful treatment outcomes. The treatment success rate (69%) in the current study participants was below the target set by WHO (>75%). Paying special attention and timely intervention in patients with high risk of unsuccessful treatment outcomes may help in improving treatment outcomes at the study site.


Combined Merkel cell carcinoma and atypical fibroxanthoma.
Although there are reports of squamous cell carcinoma arising within and adjacent to Merkel cell carcinoma, and one report of an atypical fibroxanthoma-like tumor arising in an irradiated recurrent Merkel cell carcinoma, there have previously been no reports of an immunohistochemically verified atypical fibroxanthoma occurring in conjunction with a Merkel cell carcinoma. We report on a neoplasm with distinct features of both Merkel cell carcinoma and atypical fibroxanthoma. Histologic and immunohistochemical evaluations were performed. Our results verify the finding of a combined Merkel cell carcinoma and atypical fibroxanthoma. This case is an interesting and unusual combination of tumors. The origin of the Merkel cell carcinoma is revisited. Debated origins include neuroendocrine, epithelial, and pleuripotent stem cell.


Ultrasonographic measurement of cerebral blood flow, cerebral circulation time and cerebral blood volume in vascular and Alzheimer's dementia.
Vascular dementia (VD) and Alzheimer's dementia (AD) are the most common differential diagnoses in patients with cognitive impairment. Although of different etiology, small vessel disease is postulated to be present in both conditions. We investigated global cerebral blood flow (CBF), global cerebral circulation time (CCT) and global cerebral blood volume (CBV) in VD and AD patients using a multimodal ultrasound (US) approach. 20 VD and 20 AD patients were included and compared with 12 age-matched controls. Duplex US of both internal carotid and vertebral arteries was performed to measure CBF. CCT was defined as the time delay of an echo-contrast bolus arrival between the internal carotid artery and internal jugular vein using extracranial Doppler. CBV was calculated as the product of CBF and CCT. CBF was significantly lower (VD: 570 +/- 61; AD: 578 +/- 77; controls: 733 +/- 54 ml/min) and CCT significantly longer (8.8 +/- 2.6; 8.2 +/- 1.4; 6.4 +/- 0.8 s) in both patient groups compared with controls (p < 0.003). No difference in CBF and CCT was found between the two patient groups. CBV was similar in all three groups (82 +/- 20; 79 +/- 19; 78 +/- 9 ml). The equally reduced CBF and prolonged CCT in VD and AD support the hypothesis, that small vessel disease is a relevant factor in both types of dementia. The presented multimodal US approach helps to assess the extent of changes in the global cerebral hemodynamics in patients with dementia but does not allow a differentiation between VD and AD.


Cardioprotective potency of the radical scavenger S-2-(3 aminopropylamino) ethylphosphorothioic acid in the post-ischaemic rat heart.
S-2-(3 aminopropylamino) ethylphosphorothioic acid (WR-2721) shown to surpass radical scavenging thiols in their radioprotective efficacy in cancer-type diseases has been tested for its protective potential in the reperfused heart. We investigated the radical scavenger properties of the compound in a radical generating system in vitro as well as in isolated rat hearts subjected to 30 min ischaemia and 30 min reperfusion with the electron-paramagnetic resonance spin trap technique. The action on high-energy phosphates is observed by means of phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy while its influence on left ventricular systolic segmental length change (SSLC) during 60 min reperfusion following 60 min regional ischaemia was assessed with a fibreoptic system in anaesthetized open-chest rats. WR-2721 (0.1 mM) reduced the vascular concentration of radical adduct in isolated hearts by up to 78% (275 +/- 84% of pre-ischaemic baseline values vs 1260 +/- 413%, p < 0.05) between 5 and 12.5 min reperfusion. This was accompanied by a reduction of the left ventricular end diastolic pressure to pre-ischaemic values at 30 min of reperfusion (9 +/- 6 mmHg vs 42 +/- 8 mmHg in the absence of WR-2721, p < 0.02). An accelerated recovery of creatine phosphate levels (78 +/- 5% of pre-ischaemia values vs 41 +/- 5% within 60 min reperfusion: p < 0.05) was observed under similar conditions with NMR-spectroscopy, although the post-ischaemic tissue content of adenosine triphosphate was not affected.(ABSTRACT TRUNCATED AT 250 WORDS)


Bioreactor-Based Adherent Cells Harvesting from Microcarriers with 3D Printed Inertial Microfluidics.
Harvesting adherent cells from microcarriers has become one of the major challenges of the downstream bioprocessing at large scale the current method has high maintenance and operation cost, which are the results of frequent clogging, due to cell lysing effect and microcarrier cake formation on membrane-based technology. These problems hugely impede the adaptation of microcarriers technologies in large-scale cell culture and hampered the supply of cells to the clinical need. Here, we describe two 3D printing-based methods to fabricate inertial microfluidic devices for separating adherent cells from microcarriers which overcome the above-mentioned limitations. The spiral devices are employed to separate mesenchymal stem cells from the microcarriers with 99% microcarrier removal rate and 77% cell recovery rate in one round of separation.


Quantification of Strigolactones.
Strigolactones (SLs) are a family of natural products produced by the plants as shoot branching factors and responsible for the induction of hyphal branching in arbuscular mycorrhizal (AM) fungi. They have been also used by parasitic plant seeds as stimulators of their germination as a strategy to ensure the presence of a host in the environment. For all these bioactivities, SLs have kept the attention of the researchers in the last years, increasing the number of published papers, and have opened new areas of research in the multiple roles that they play in the rhizosphere and as plant hormones. However, the low amount of them produced by plants and their rapid degradability make it crucial to develop fast analytical methods with very low limits of quantification. Herein, it is described a protocol for the development of an LC-MS/MS method for the quantification of SLs, using GR24 as IS, in roots exudates and extracts.


Antibiotic resistance in bacteria from shrimp farming in mangrove areas.
Shrimp farming is a sufficiently large and mature industry to have an effective range of antimicrobial agents for most bacterial diseases in shrimp culture. However, at present, there exists great concern over the widespread use of antibiotics in aquaculture, which may result in residue of antibiotics in water and mud, and subsequently, the development of antibiotic resistance in bacteria in the environment. There is limited understanding about the effect of antibiotic residues on bacteria resistance in shrimp farming environment. Therefore, a study was conducted to investigate bacterial resistance to Norfloxacin (NFXC), Oxolinic Acid (OXLA), Trimethoprim (TMP) and Sulfamethoxazole (SMX), which were found in four shrimp farming locations in mangrove areas in Vietnam. Findings indicate that there is a relatively high incidence of bacteria resistance to these antibiotics observed in most of the studied sites, particularly to antibiotics with concentration of 0.1 microg/ml. Yet the relation between concentration of antibiotic residues and incidence of antibiotic resistance is not clearly defined. Among individual antibiotics, the incidence of resistance to TMP and SMX was higher than the others. Identification of bacteria isolated from mud samples by DNA analyzer shows that Bacillus and Vibrio are predominant among bacteria resistant to the antibiotics. The result of the study also indicates that these antibiotics in media degraded more rapidly due to the presence of resistant bacteria.


Relations between vocabulary development and verbal short-term memory: The relative importance of short-term memory for serial order and item information.
Although many studies have shown an association between verbal short-term memory (STM) and vocabulary development, the precise nature of this association is not yet clear. The current study reexamined this relation in 4- to 6-year-olds by designing verbal STM tasks that maximized memory for either item or serial order information. Although empirical data suggest that distinct STM processes determine item and serial order recall, these were generally confounded in previous developmental studies. We observed that item and order memory tasks were independently related to vocabulary development. Furthermore, vocabulary development was more strongly associated with STM for order information in 4- and 6-year-olds and with STM for item information in 5-year-olds. These data highlight the specificity of verbal STM for serial order and item information and suggest a causal association between order STM processes and vocabulary development, at least in 4- and 6-year-olds.


Sulbactam-based therapy for Acinetobacter baumannii infection: a systematic review and meta-analysis.
A number of studies have reported on the effectiveness of sulbactam-based therapies for Acinetobacter baumannii infection; however, there is little evidence that sulbactam-based therapies are more or less effective than alternative therapies. Unfortunately, there is a distinct lack of high quality data (i.e., from randomized controlled trials) available on this issue. Therefore, we conducted a systematic review and meta-analysis comparing the efficacy of sulbactam-based and non-sulbactam-based regimens in the treatment of A. baumannii infection. We searched PubMed, MEDLINE, Biomedical Central, Google Scholar, the China National Knowledge Infrastructure, the Cochrane library, and the Directory of Open Access using the terms "sulbactam and baumannii" or "maxtam and baumannii". Randomized controlled trials, controlled clinical studies, and cohort studies were considered for inclusion. The primary outcome was the clinical response rate for sulbactam-based therapy vs comparator therapies. Four studies (1 prospective, 3 retrospective) were included in the meta-analysis. Sulbactam was given in combination with ampicillin, carbapenem, or cefoperazone (n=112 participants). Comparator drugs included colistin, cephalosporins, anti-pseudomonas penicillins, fluoroquinolones, minocycline/doxycycline, aminoglycosides, tigecycline, polymyxin, imipenem/cilastatin, and combination therapy (n=107 participants). The combined clinical response rate odds ratio did not significantly favor sulbactam-based therapy over comparator therapy (odds ratio=1.054, 95% confidence interval=0.550-2.019, p=0.874), nor did any of the individual study odds ratios. The available evidence suggests that sulbactam-based therapy may be similarly efficacious to alternative antimicrobial therapies for the treatment of A. baumannii infection. Further research on this issue is warranted given the limited availability of data from high quality/randomized controlled trials.


The Effect of Povidone-Iodine Nasal Spray on Nasopharyngeal SARS-CoV-2 Viral Load: A Randomized Control Trial.
To determine the effect of povidone-iodine (PVP-I) nasal sprays on nasopharyngeal (NP) viral load as assessed by cycle threshold (Ct) on quantitative polymerase chain reaction (qPCR) of SARS-CoV-2 in outpatients. Three arm, triple blinded, randomized, placebo-controlled clinical trial. Participants were randomized within 5 days of testing positive for COVID-19 to receive nasal sprays containing placebo (0.9% saline), 0.5% PVP-I, or 2.0% PVP-I. NP swabs for qPCR analysis were taken at baseline, 1-hour post-PVP-I spray (two sprays/nostril), and 3 days post-PVP-I spray (20 sprays/nostril). Symptom and adverse event questionnaires were completed at baseline, day 3, and day 5. University of Pennsylvania Smell Identification Tests (UPSIT) were completed at baseline and day 30. Mean Ct values increased over time in all groups, indicating declining viral loads, with no statistically significant difference noted in the rate of change between placebo and PVP-I groups. The 2.0% PVP-I group showed statistically significant improvement in all symptom categories; however, it also reported a high rate of nasal burning. Olfaction via UPSIT showed improvement by at least one category in all groups. There were no hospitalizations or mortalities within 30 days of study enrollment. Saline and low concentration PVP-I nasal sprays are well tolerated. Similar reductions in SARS-CoV-2 NP viral load were seen over time in all groups. All treatment groups showed improvement in olfaction over 30 days. These data suggest that dilute versions of PVP-I nasal spray are safe for topical use in the nasal cavity, but that PVP-I does not demonstrate virucidal activity in COVID-19 positive outpatients. 2 Laryngoscope, 132:2089-2095, 2022.


Mapping of the mouse serum amyloid A gene cluster by long-range polymerase chain reaction.
The present study defines the organization of the mouse serum amyloid A (Saa) gene cluster on chromosome 7. A polymerase chain reaction (PCR)-based strategy was used successfully to generate a complete map of the mouse Saa genes, defining a linkage group of 3'-Saa2-5'/5'-Saa1-3'/5'-Saa4-3'/5'-Saa5-3'/5'-+ ++Saa3-3', with a maximum size of 45 kilobases (kb). This contrasts with the 150 kb human SAA gene cluster, which has been previously defined. The tight linkage of both mouse Saas and human SAAs is of potential functional significance, since the genes that encode the acute phase serum amyloid A proteins are known to exhibit co-ordinate transcriptional regulation. The present results thus suggest that selective pressure may exist which maintains the co-ordinately transcribed Saa genes in close physical proximity. This study, furthermore, demonstrates the utility of a novel PCR-based approach for fine mapping of tightly clustered linkage groups. The strategy used possesses a number of advantages over previously described techniques, such as long-range restriction mapping, since it facilitates the concurrent determination of not only precise relative map positions, but also the relative transcriptional orientations of assayed paired loci. Although presently limited in resolution to genes not more than 27 kb apart, future technical advances are likely to extend the applicability of this approach in mapping experiments to less tightly linked clusters of genes.


Organ culture of human articular cartilage: studies on sulphated glycosaminoglycan synthesis.
An organ culture system is described for adult human articular cartilage obtained from joints after femoral head replacement operations. Cartilage slices maintain maximal viability for 2 days in culture as assessed by uptake of [3H]uridine and [3H]leucine into whole tissue, and 35SO4 into sulphated glycosaminoglycans (GAGs). Since GAGs are the components of cartilage matrix, the depletion of which is associated with osteoarthrosis, a method for measuring sulphated GAG synthesis in culture has been investigated.


Investigation of potential early Histologic markers of pediatric inflammatory bowel disease.
Early manifestations of pediatric inflammatory bowel disease (IBD) can be relatively nonspecific. Initial mucosal biopsies may not be conclusive, delaying the diagnosis until subsequent biopsies demonstrate typical histologic features of IBD. We hypothesized that certain inflammatory cell types may be utilized as early histologic indicators of IBD in children. A retrospective analysis compared histologic findings from initially inconclusive or negative endoscopic studies in 22 patients who were subsequently diagnosed with IBD (after diagnostic endoscopy) to those of 20 comparison patients with functional abdominal pain matched for age, gender, and study type. A pediatric pathologist, blinded to study group, reviewed biopsies for histologic abnormalities. Eosinophil densities were obtained from the stomach, duodenum, and rectosigmoid areas. Immunohistochemistry (IHC) staining for tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-9 (MMP-9) was performed on the stomach and rectosigmoid areas. Gastritis and colonic crypt distortion were present in the IBD group at a greater rate (61 % vs. 22 %, p = 0.020; 34 % vs. 4 %, p = 0.008, respectively). Peak and mean eosinophil densities in the rectosigmoid area were greater in the IBD group (17.0/hpf vs. 5.0/hpf, p = 0.0063; 12.3/hpf vs. 4.2/hpf, p = 0.0106, respectively). TNF-α and MMP-9 staining did not reveal any significant differences. Our data suggests that significantly greater inflammation in the stomach, crypt distortion in the colon, and eosinophilia in the rectosigmoid distinguished the IBD group from the comparison group at the time of the initial endoscopic evaluation.


Systemwide effects of ER-intracellular membrane contact site disturbance in primary endothelial cells.
Membrane contact sites (MCS) make up a crucial route of inter-organelle non-vesicular transport within the cell. Multiple proteins are involved in this process, which includes the ER-resident proteins vesicle associated membrane protein associated protein A and -B (VAPA/B) that form MCS between the ER and other membrane compartments. Currently most functional data on VAP depleted phenotypes have shown alterations in lipid homeostasis, induction of ER stress, dysfunction of UPR and autophagy, as well as neurodegeneration. Literature on concurrent silencing of VAPA/B is still sparse; therefore, we investigated how it affects the macromolecule pools of primary endothelial cells. Our transcriptomics results showed significant upregulation in genes related to inflammation, ER and Golgi dysfunction, ER stress, cell adhesion, as well as Coat Protein Complex-I and -II (COP-I, COP-II) vesicle transport. Genes related to cellular division were downregulated, as well as key genes of lipid and sterol biosynthesis. Lipidomics analyses revealed reductions in cholesteryl esters, very long chain highly unsaturated and saturated lipids, whereas increases in free cholesterol and relatively short chain unsaturated lipids were evident. Furthermore, the knockdown resulted in an inhibition of angiogenesis in vitro. We speculate that ER MCS depletion has led to multifaceted outcomes, which include elevated ER free cholesterol content and ER stress, alterations in lipid metabolism, ER-Golgi function and vesicle transport, which have led to a reduction in angiogenesis. The silencing also induced an inflammatory response, consistent with upregulation of markers of early atherogenesis. To conclude, ER MCS mediated by VAPA/B play a crucial role in maintaining cholesterol traffic and sustain normal endothelial functions.


Treatment of airborne asbestos and asbestos-like microfiber particles using atmospheric microwave air plasma.
Atmospheric microwave air plasma was used to treat asbestos-like microfiber particles that had two types of ceramic fiber and one type of stainless fiber. The treated particles were characterized via scanning electron microscopy (SEM) and X-ray diffraction (XRD). The experiment results showed that one type of ceramic fiber (Alumina:Silica=1:1) and the stainless fiber were spheroidized, but the other type of ceramic fiber (Alumina:Silica=7:3) was not. The conversion of the fibers was investigated by calculating the equivalent diameter, the aspect ratio, and the fiber content ratio. The fiber content ratio in various conditions showed values near zero. The relationship between the normalized fiber vanishing rate and the energy needed to melt the particles completely per unit surface area of projected particles, which is defined as η, was examined and seen to indicate that the normalized fiber vanishing rate decreased rapidly with the increase in η. Finally, some preliminary experiments for pure asbestos were conducted, and the analysis via XRD and phase-contrast microscopy (PCM) showed the availability of the plasma treatment.


A novel method for creating endodontic access preparations through all-ceramic restorations: air abrasion and its effect relative to diamond and carbide bur use.
Access through porcelain restorations is a technically delicate and stressful procedure. Although this is a common dilemma in endodontics, little research has explored alternatives in cutting through porcelain. The purpose of this study was to compare the use of a carbide bur plus water, diamond bur plus water, and air abrasion to access through porcelain. All-ceramic samples were accessed using the different techniques. Samples were evaluated using two transillumination methods, white light, and fluorescent liquid penetrant described by the American Society for Testing and Materials. Edge chipping, microcracking, and catastrophic fracture of porcelain caused by the techniques were statistically compared. Fluorescent liquid penetrant was a more sensitive method for microcrack detection. There were significant differences between the preparation techniques. Air abrasion was significantly less destructive, and caused no catastrophic fractures, edge chipping or microcracks. Preparation by air abrasion took longer to complete.


Novel imidazopyrimidines-based molecules induce tetramerization of tumor pyruvate kinase M2 and exhibit potent antiproliferative profile.
Discovery of novel and potent lead molecules for the specific therapeutic targets by de novo drug design is still in infancy. Here, we disclose the unprecedented development of imidazopyri(mi)dine-based tumor pyruvate kinase M2 (PKM2) modulators by subsequent link and grow strategy. The most potent modulator 15n acts as a PKM2 activator with an AC50 of 90 nM, with considerable cancer cell-selectivity and membrane-permeability. NMR metabolomics studies also revealed that treatment with 15n results in diminution in lactate concentrations in MCF-7 cells. 15n binds to a previously reported site at PKM2 adjacent to the interface of two monomers. In molecular dynamics (MD) simulation studies, it was observed that 15n stabilizes the PKM2 at the dimeric interface, assisting in the formation of a biologically active tetramer conformation. 15n was also screened on MCF-7 breast cancer cell lines grown on 3-D scaffolds, and the results exhibited better anticancer potential compared to control, paving the way for future clinical studies.


Pregnancy associated thrombotic thrombocytopenic purpura: Practical issues for patient management.
Thrombotic thrombocytopenic purpura (TTP) is a severe, life-threatening disease that needs urgent diagnosis and prompt therapeutic intervention. In pregnant women TTP may complicate the course of gestation putting mother and child at vital risk. Differential diagnosis with other obstetric and medical disorders may be difficult due to the overlap of several clinical and laboratory findings. Our understanding of the pathophysiology of TTP has allowed ADAMTS13 testing to have a central role in confirming the clinical diagnosis but the main limitation is that an ADAMTS13 assay is not available in "real time". Here we report the clinical course and treatment outcome of two young women with clinical manifestations of pregnancy associated TTP and briefly discuss the main topics of disease diagnosis and management.


Male involvement in the use of family planning and associated factors in Gondar City, Northwest Ethiopia: A community-based cross-sectional study.
To assess the level of male involvement in the use of family planning and associated factors among married men. A community-based cross-sectional study was conducted from March to April 2018. Married men were randomly selected from four randomly selected kebeles, the smallest administrative division in Ethiopia. Data were collected using an interviewer-administered structured questionnaire. A multivariable logistic regression model was fitted to identify the associated factors with male involvement in the use of family planning. A total of 741 of 744 (99.6%) married men responded to the interview. The overall involvement of men in family planning was 70%. Men aged 25-30 years (adjusted odds ratio [AOR] 3.78, 95% confidence interval [CI] 1.67-8.54), attending secondary school education (AOR 2.25, 95% CI 1.173-4.31), attending college and higher education (AOR 7.49, 95% CI 3.87-14.51), had more than five children (AOR 14.6, 95% CI 4.37-48.97), and had 3-4 children (AOR 16.13, 95% CI 6.31-41.16) were the factors significantly associated with male involvement in family planning. The involvement of married men in family planning was low. Encouraging the education of younger men by considering those who do not have any children is an essential measure to enhance men's involvement in family planning.


Bioconversion of sitosterol to useful steroidal intermediates by mutants of Mycobacterium fortuitum.
A series of mutants which are blocked at various stages of the sterol degradative pathway have been isolated from the potent sterol degrader Mycobacterium fortuitum ATCC-6842. Sitosterol bioconversions by these mutants result in the accumulation of a number of intermediate compounds, some of which are potentially useful as substrates in the manufacture of medically important steroids. These intermediates include androst-4-ene-3,17-dione, androsta-1,4-diene,3,17-dione, ring A-degraded tricyclic compounds and various 9alpha-hydroxy-steroids.


Column purification of adenovirus cores.
A simple and efficient technique for purifying adenoviral chromatin (nucleoprotein cores) with Sephacryl S-1000 is described. This method is significantly faster than previous methods and gives a higher degree of purity with an increased recovery of the nucleoprotein. In addition, the structural integrity of the cores is maintained.


Pain development and consumption of analgesics after oral surgery in relation to personality characteristics.
In the present study a survey has been performed of the pain development and analgesic intake in 100 patients following elective oral surgery of impacted third molars in relation to preoperative assessment of personality characteristics. Our results show that no sex differences existed preoperatively concerning personality characteristics, concerning postoperative pain development or analgesic consumption. Fourteen patients reported no pain at all and 40 patients did not use any analgesics in the postoperative period. We also found a good correlation between total sum of pain scores and analgesic intake. Patients undergoing surgery in the morning reported a lower total sum of pain scores, reported pain at fewer occasions and tended to require less analgesics than patients being subjected to surgery in the afternoon. The patients who reported a total sum of pain scores in the upper percentile during the postoperative period rated their general health worse, as rated on the General Health (GH) questionnaire, and used more analgesics than did patients in the lower percentile. Also, patients not using any analgesics reported less symptoms of distress according to the GH scale as compared to patients using analgesics. In general, however, less than 10% of the variance in postoperative pain and consumption of analgesics could be explained by the preoperative factors studied.


Analysis of outcome using a levator sparing technique of abdominoperineal excision of rectum and anus. Cylindrical ELAPE is not necessary in all patients.
Abdominoperineal excision of rectum (APE) for cancer has a higher rate of local recurrence with a poorer outcome than stage matched anterior resection. The cylindrical excision (ELAPE) has been advocated to reduce local recurrence. However, this operation has greater morbidity and requires more post operative care. We report our outcomes from a single centre using a levator sparing dissection. All patients undergoing APE from January 2007-June 2011 were evaluated. Case notes operation notes and pathology results were reviewed for complications and staging. Follow-up data for survival and recurrence were obtained from the cancer registry, imaging and from clinic follow up. Of all rectal cancers (n = 361), 43 had APE with curative intent. Median age was 67(IQR 59-76). Median tumour height was two centimetres from the dentate line (IQR 1-3.5 cm). Neoadjuvant chemoradiotherapy was given in 98% of APE resections with curative intent. Median post operative hospital stay was 10 days (8-15). At a median follow up of 38 months (IQR30-49) for patients undergoing curative resection, 2 patients (4.6%) had local recurrence and overall mortality was 18.6% (n = 8). With adequate neoadjuvant chemoradiotherapy, a levator sparing excision of rectum remains a safe option with less morbidity and perioperative complications than has been described for ELAPE.


Wet and dry deposition of Chernobyl releases.
The passage of the Chernobyl plume over the United Kingdom in May 1986 led to the deposition of radionuclides on the ground by wet and dry deposition processes. Here we analyse rainfall during the passage of the plume and the published monitoring data obtained afterwards, and show that levels of deposited 137Cs can be closely related to rainfall intercepting the plume. 137Cs was present in the atmosphere mostly as particulate species with wet deposition mechanisms dominating. In contrast, 131I was present as particulate and vapour phase material, and reported levels on grass and in cow's milk show that both wet and dry deposition mechanisms were important. 131I on grass and in cow's milk therefore shows a different geographic pattern to 137Cs, and is not so closely related to rainfall. We have calculated washout factors for locations where there are data on deposition, rainfall and air concentrations during the passage of the Chernobyl plume. From these factors and interpolated concentrations in the atmosphere, the total deposition of 137Cs has been estimated at each of 4,000 rain gauge stations in the United Kingdom. The results are presented as deposition contours and have been compared with measurements in parts of the country. Estimates of the total deposition of 131I and 137Cs show that less than or equal to 1% of the estimated total releases from Chernobyl were deposited on the United Kingdom.


Outcomes of preoperative bevacizumab in diabetics with nonclearing vitreous hemorrhage without tractional detachment - A quasi-randomized retrospective study.
To assess whether preoperative bevacizumab (BVZ) in treatment-naïve eyes with proliferative diabetic retinopathy (PDR) and vitreous hemorrhage (VH) without tractional retinal detachment (TRD) leads to lesser macular edema and better visual outcome compared to eyes that do not receive BVZ. This quasi-randomized retrospective study included 217 treatment-naïve eyes with nonclearing VH without TRD that had vitrectomy with or without BVZ and had a minimum 6-months follow-up. Postoperative variables, including visual acuity (BCVA), central macular thickness (CMT) at 1 month, and need for additional anti-VEGF injections till 6 months follow-up, were recorded for analysis. Of the 217 eyes, 107 eyes (49%) received preoperative BVZ and 110 (51%) did not. Groups were comparable in terms of preoperative characteristics. At 1 month, mean CMT was significantly higher in eyes without BVZ (310 ± 33 m vs. 246 ± 34m; P < 0.001). The likelihood of developing center-involving DME at 1 month after vitrectomy was 67% lower if the eye received preoperative BVZ (OR = 0.33, 95%CI = 0.18-2.54, P = 0.56). Though BCVA improved significantly in both groups at 1 month, it was 1/3rd of a line better in the BVZ group (b coefficient = -0.035 logMAR, 95%CI = -0.04 to -0.008 logMAR, P = 0.01). Preoperative BVZ in treatment-naïve eyes with PDR and VH but without TRD lead to better macular status and marginally improved vision at 1 month, which was maintained at 6 months. In view of these results, patients may be offered BVZ only when it is readily affordable to them.


Ancient DNA extraction from soils and sediments.
DNA contained in soils and sediments can provide novel insights into past environments and ecosystems. In this chapter, I describe an efficient and effective technique to extract total DNA from sediments in a manner that minimizes the coextraction of PCR-inhibitory compounds. I describe two different approaches: one that is suitable for large (up to 10 g wet weight) amounts of substrate, and a second that is more appropriate for small (up to 0.5 g) amounts of substrate. Finally, I discuss some of the obstacles that may be encountered in the process of extracting DNA from soils and sediments and suggest approaches to circumvent some common problems.


Exosomal miR-181a-5p reduce Mycoplasma gallisepticum (HS strain) infection in chicken by targeting PPM1B and activating the TLR2-mediated MyD88/NF-κB signaling pathway.
Mycoplasma gallisepticum (MG) is one of the most important pathogens that causes chronic respiratory disease (CRD) in chickens. Exosomes secreted from cells have been well demonstrated to deliver miRNAs to recipient cells to modulate cellular functions. The purpose of this study is to explore the underlying functions and mechanisms of exosomal miR-181a-5p in MG-HS infection. In this study, we found that miR-181a-5p expression in vivo and in vitro was significantly up-regulated after MG-HS infection. It was also upregulated in exosomes, which were derived from MG-HS-infected type-II pneumocytes cells (CP-II). In addition, exosomes secreted by MG-HS-infected CP-II were able to transfer miR-181a-5p to recipient chicken embryo fibroblast cells (DF-1), resulting in a significant upregulation of miR-181a-5p expression in recipient DF-1 cells. We further identified that Mg2+/Mn2+-dependent protein phosphatase 1B (PPM1B) was the target gene of miR-181a-5p. Overexpression of miR-181a-5p or knockdown of PPM1B activated the nuclear factor-κB (NF-κB) signaling pathway, whereas inhibition of miR-181a-5p and overexpression of PPM1B led to the opposite results. Besides, up-regulation of miR-181a-5p significantly increased the expression of toll-like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88), tumor necrosis factors alpha (TNF-α) and interleukin-1β (IL-1β), whereas inhibition of miR-181a-5p showed a contrary result. Up-regulation of miR-181a-5p promoted cell proliferation, cell cycle progression and inhibited apoptosis to resist MG-HS infection. Moreover, overexpression of miR-181a-5p significantly negative regulated the expression of Mycoplasma gallisepticum adhesin protein (pMGA1.2) by directly inhibiting PPM1B. Thus, we concluded that exosomal miR-181a-5p from CP-II cells activated the TLR2-mediated MyD88/NF-κB signaling pathways by directly targeting PPM1B to promote the expression of pro-inflammatory cytokines for defending against MG-HS infection in recipient DF-1 cells.


Determination of the Length of Pancreatic Ductotomy by Pancreaticoscopy During Frey's Procedure for Chronic Pancreatitis.
To study the impact of pancreaticoscopy during Frey's procedure for treating chronic pancreatitis (CP). Excision of the central part of the head of the pancreas along with longitudinal pancreaticotomy (Frey's procedure) is widely performed for the treatment of CP. However, there is no reliable method to determine the necessary length of longitudinal pancreaticotomy during surgery for CP. Thirty-five consecutive patients with CP were scheduled for Frey's procedure with intraoperative pancreaticoscopy. The length of the longitudinal pancreaticotomy was tailored by pancreaticoscopy in the following manner: (1) it did not extend beyond the neck in case of a uniformly dilated main duct with patent branch duct confluences and a clear lumen; (2) in case of an obstructed main duct or branch duct confluence of any cause, the main duct was opened to include the most distal obstruction. All patients underwent Frey's procedure and intraoperative pancreaticoscopy. Based on the pancreaticoscopy findings, pancreaticotomy over the body of the gland was not necessary in 34% of the patients. A short (4-6 cm) ductotomy extension over the pancreatic body was required in 14% of the patients. Full-length pancreaticotomy was required in 52% of the patients. The median operative time was 145 minutes, and the median blood loss was 70 mL. Four patients (11.4%) experienced postoperative complications. There were no 90-day postoperative mortality or hospital readmission rates. At the median follow-up of 19 months, 31 patients (88.5%) had no pain attacks requiring medication. Intraoperative pancreaticoscopy helps to determine the length of longitudinal pancreaticotomy and reduce pancreatic trauma during Frey's procedure for treating CP.


MicroRNA roles in signalling during lactation: an insight from differential expression, time course and pathway analyses of deep sequence data.
The study examined microRNA (miRNA) expression and regulatory patterns during an entire bovine lactation cycle. Total RNA from milk fat samples collected at the lactogenesis (LAC, day1 [D1] and D7), galactopoiesis (GAL, D30, D70, D130, D170 and D230) and involution (INV, D290 and when milk production dropped to 5 kg/day) stages from 9 cows was used for miRNA sequencing. A total of 475 known and 238 novel miRNAs were identified. Fifteen abundantly expressed miRNAs across lactation stages play regulatory roles in basic metabolic, cellular and immunological functions. About 344, 366 and 209 miRNAs were significantly differentially expressed (DE) between GAL and LAC, INV and GAL, and INV and LAC stages, respectively. MiR-29b/miR-363 and miR-874/miR-6254 are important mediators for transition signals from LAC to GAL and from GAL to INV, respectively. Moreover, 58 miRNAs were dynamically DE in all lactation stages and 19 miRNAs were significantly time-dependently DE throughout lactation. Relevant signalling pathways for transition between lactation stages are involved in apoptosis (PTEN and SAPK/JNK), intracellular signalling (protein kinase A, TGF-β and ERK5), cell cycle regulation (STAT3), cytokines, hormones and growth factors (prolactin, growth hormone and glucocorticoid receptor). Overall, our data suggest diverse, temporal and physiological signal-dependent regulatory and mediator functions for miRNAs during lactation.


Effects of three resin monomers on the cellular glutathione concentration of cultured human gingival fibroblasts.
Oral and systemic cells are permanently exposed to various types of xenobiotics, such as dental restorative materials, which may subsequently cause adverse effects. Objective of the present investigation was to analyze the effects of three important resin monomers on the glutathione metabolism of human gingival fibroblasts after an incubation period of 4h. Cells were exposed to various concentrations of 2-hydroxyethyl methacrylate (HEMA; 0.1-10 mM), triethylene-glycol dimethacrylate (TEGDMA; 0.05-2.5 mM), and urethane dimethacrylate (UDMA; 0.005-0.25 mM). Subsequently, cellular glutathione (GSH) concentrations were determined after a treatment period of 4h using the monobromobimane assay. Data were statistically evaluated using Tukey ANOVA with p<0.05. GSH depletion was dependent on the type of the resin monomer: UDMA>TEGDMA>HEMA. The concentrations for a 50%-reduction of cellular GSH varied between 0.1 mM (0.05 mM) (UDMA), 0.33 mM (0.09 mM) (TEGDMA), and 1.6 mM (0.8 mM) (HEMA). Simultaneously, no decrease of cell numbers was found at any tested concentration. These data indicate that the investigated resins may cause cell damage due to depletion of intracellular GSH level even at low concentrations within a short period of time. The decrease of GSH is an early reaction, which is triggered prior to other cytotoxic alterations.


[Numerical analysis on the influence factors of sample stacking in capillary electrophoresis].
Sample stacking in capillary electrophoresis can concentrate sample species through the electrical field strength gradient caused by the inhomogeneous buffer concentration field in capillary. The factors that affect the sample stacking process have been analyzed in detail by using a 1-D mathematic model. It was found from the simulation results that the electrical charge number and the electrical charge sign of sample particles can affect the electrophoretic velocity, which in turn has an important influence on the stacking process. The electrical potential can affect the migration captime of sample particles to reach detection window, and the initial length of sample plug has significant influence on the maximal sample concentration after stacking and the time to get the optimal stacking effect. The results obtained are helpful to the improvement of the sample stacking technique.


"Horseshoe kidney" in an ectopic position. A case report.
We present a case study of a horseshoe kidney, harvested from a 68-year-old patient who died suddenly from rupture of an aneurysm of the ascending aorta. The kidney was ectopically placed, its isthmus corresponding to the terminal aorta. The two kidneys were unequal in size, the right kidney being larger than the left. We discovered four renal arteries, two on the right and two on the left side, and four renal veins, one left, one middle and two on the right side. The isthmus was blood supplied by the left and the right inferior renal arteries while the venous blood was collected by the left and middle renal veins.


[Comparative study of the effects of 2 types of protein-energy malnutrition followed by a balanced refeeding, on the lipase, phospholipase A2 and amylase activities of the pancreas and pancreatic juice in the growing rat].
The intake of 2 p. 100 casein diet as only protein source decreased overall phospholipase A2 activity. Amylase activity was more affected than lipase one. The effects of intake of 5 p. 100 gluten diet as only protein source were less important than 2 p. 100 casein diet. Refeeding on 20 p. 100 or 15 p. 100 casein diet caused a considerable increase of phospholipase A2, lipase and amylase activities.


Study of Morphological Changes in Rat Liver Caused by Occlusion of Inferior Vena Cava.
Up to date, there are no reports on animal survival rate and morphological changes in the liver caused by the impairment of blood outflow from the liver and its time dependence. Moreover, the impact of duration and degree of occlusion of inferior vena cava on pathological changes was not investigated yet. This study aimed at the assessment of the survival rate and morphological changes in the liver with varying degrees of occlusion of inferior vena cava. The exact timing of the reversibility of pathological processes was determined. Rats (n = 160) were randomly divided into five groups: I - control group (CG) (n = 20); II - sham group (SG) (n = 20); III - intervention group (IG-1) (narrowing of the lumen of the inferior vena cava by 25%) (n = 40); IV intervention group (IG-2) (narrowing by 50%) (n = 40); and intervention group V (IG-3) (narrowing by 75%) (n = 40). The level of postoperative pain, the body and liver weight of the animals, histological examination, morphometry, and macroscopic evaluation of abdominal organs were carried out on the 1, 3, 7, 14, and 30 days following the surgical intervention. The survival rate of animals was assessed using the Kaplan-Meier method. On the 30th day, the rat grimace scale indices in the IG-1 (P ≤ 0.05), IG-2, and IG-3 (P ≤ 0.001) groups were higher. By body weight, this indicator on the 30th day was lower in the IG-1 (P ≤ 0.05), IG-2, and IG-3 (P ≤ 0.001) groups compared to the CG and SG groups. In the IG1 and IG2 groups, the survival rates were 72.5% and 65.0%, respectively. The lowest survival rate was observed in the IG3 group (22.5%). Compression of the inferior vena cava by 75% led to an increase in animal mortality and the development of persistent morphological changes in the liver. At the same time, the survival rate of animals and the extent of changes in the liver with narrowing of the inferior vena cava by 25% and 50% had similar results. The results acquired possess scientific and practical importance.


Trends in Obesity Among Adults in the United States, 2005 to 2014.
Between 1980 and 2000, the prevalence of obesity increased significantly among adult men and women in the United States; further significant increases were observed through 2003-2004 for men but not women. Subsequent comparisons of data from 2003-2004 with data through 2011-2012 showed no significant increases for men or women. To examine obesity prevalence for 2013-2014 and trends over the decade from 2005 through 2014 adjusting for sex, age, race/Hispanic origin, smoking status, and education. Analysis of data obtained from the National Health and Nutrition Examination Survey (NHANES), a cross-sectional, nationally representative health examination survey of the US civilian noninstitutionalized population that includes measured weight and height. Survey period. Prevalence of obesity (body mass index ≥30) and class 3 obesity (body mass index ≥40). This report is based on data from 2638 adult men (mean age, 46.8 years) and 2817 women (mean age, 48.4 years) from the most recent 2 years (2013-2014) of NHANES and data from 21,013 participants in previous NHANES surveys from 2005 through 2012. For the years 2013-2014, the overall age-adjusted prevalence of obesity was 37.7% (95% CI, 35.8%-39.7%); among men, it was 35.0% (95% CI, 32.8%-37.3%); and among women, it was 40.4% (95% CI, 37.6%-43.3%). The corresponding prevalence of class 3 obesity overall was 7.7% (95% CI, 6.2%-9.3%); among men, it was 5.5% (95% CI, 4.0%-7.2%); and among women, it was 9.9% (95% CI, 7.5%-12.3%). Analyses of changes over the decade from 2005 through 2014, adjusted for age, race/Hispanic origin, smoking status, and education, showed significant increasing linear trends among women for overall obesity (P = .004) and for class 3 obesity (P = .01) but not among men (P = .30 for overall obesity; P = .14 for class 3 obesity). In this nationally representative survey of adults in the United States, the age-adjusted prevalence of obesity in 2013-2014 was 35.0% among men and 40.4% among women. The corresponding values for class 3 obesity were 5.5% for men and 9.9% for women. For women, the prevalence of overall obesity and of class 3 obesity showed significant linear trends for increase between 2005 and 2014; there were no significant trends for men. Other studies are needed to determine the reasons for these trends.


ADAMTS-7 promotes vascular smooth muscle cells proliferation in vitro and in vivo.
Vascular smooth muscle cell (VSMC) proliferation and migration are pivotal for the pathogenesis of atherosclerosis and post-angioplasty restenosis. We have recently reported that a disintegrin and metalloproteinase with thrombospondin motifs-7 (ADAMTS-7), a novel metalloproteinase, contributes directly to neointima formation by mediating VSMC migration. However, whether ADAMTS-7 affects VSMC proliferation remains unclear. In this study, we found that luminal adenoviral delivery of ADAMTS-7 aggravated intimal hyperplasia 7 d after injury, paralleled by an increased percentage of PCNA-positive cells in both intima and media. In contrast, perivascular administration of ADAMTS-7 siRNA, but not scrambled siRNA to injured arteries attenuated intimal thickening at day 7, paralleled with reduced intimal VSMC replication, without alteration of VSMC proliferation in the media. In accordance, [(3)H]-thymidine incorporation assay in primary cultured rat VSMCs revealed an enhanced replication rate (by 61%) upon ADAMTS-7 overexpression and retarded proliferation (by 23%) upon ADAMTS-7 siRNA administration. Our data demonstrates that ADAMTS-7 promotes VSMC proliferation both in vitro and in vivo. ADAMTS-7 may therefore serve as a novel therapeutic target for atherosclerosis and post-angioplasty restenosis.


ALT (allogeneic limbal transplantation): a new surgical technique for limbal stem cell deficiency.
Limbal stem cell deficiency (LSCD) is a rare but extremely relevant disease of the eye. LSCD patients often require a variety of surgical procedures, including keratoplasty in some cases. However, the outcome of these surgeries, including opacification and revascularization, is often frustrating due to LSCD relapse. We developed a new surgical technique for the treatment of LSCD in which partial allogenic limbal transplantation (ALT) is carried out as part of penetrating keratoplasty (PK). After the PK, 1-8 slices from the limbal tissue of the donor graft are prepared and placed under the double running sutures attaching the corneal graft. This procedure was performed on 14 patients with LSCD, caused by severe ocular burn in 5 cases and by infection in 9. Between one and eight limbal transplants were used depending on the extension of the LSCD. All 14 patients showed stable or increased visual acuity after the ALT surgery compared to their preoperative visual acuity. All of the grafts were integrated into the superficial corneal layers without progression of corneal vascularization beyond the limbal grafts. The median follow-up period was 12 months on average. The ALT method seems to be a promising surgical procedure for the treatment of patients with LSCD. It can be properly carried out in the context of keratoplasty and does not require a separate donor tissue. The ALT grafts may offer the possibility of constructing a new limbal region, resulting in stable or even increased visual acuity and the absence of corneal vascularization.


Rehabilitation vs drug therapy for urge urinary incontinence: short-term outcome.
The objective of this study was to compare the short-term effectiveness of rehabilitation treatment with a standard drug treatment for urge urinary incontinence (UUI). The study design includes parallel clinical trial in an outpatient urogynecologic clinic setting. The subjects were 44 women who suffered from UUI and who were systematically assigned to a rehabilitation group (REH) (N=24) or a medication group (MED) (N=20). The intervention for REH was consisted of five visits during a 3-month period of pelvic floor muscle training and behavioral training, whereas for MED was extended release oxybutynin at 5 mg/day, for 3 months. The urinary symptoms considered were frequency of voiding per day and night (freq/day and freq/night), number of incontinent episodes per week based on a bladder diary, and data based on the Incontinence Quality of Life Instrument (I-QoL). In the within-group comparison, both groups had improved significantly over time with respect to urinary symptoms and I-QoL (p<0.01). In addition, there was a significant group-time interaction effect on freq/day. While REH improved during the 3-month follow-up period, the MED group deteriorated to mean baseline value (p<0.01). A significant negative association was found between the urinary symptoms and the I-QoL at the end of follow-up (r (p)=-0.35 to -0.62, p<0.05). Three months after the intervention, both groups maintained the achievements of the intervention period. In addition, the REH group demonstrated additional improvement in mean freq/day while the condition of MED patients deteriorated to baseline values.


Screening for coeliac disease among Egyptian children.
To screen for coeliac disease in Egyptian children with non-endocrinal short stature, refractory iron deficiency anaemia and type 1 diabetes. Also, to evaluate the sensitivity and specificity of different serological tests for diagnosis of coeliac disease (CD). The study included 292 patients with clinical risk of CD. Testing for coeliac antibodies was performed, together with upper gastrointestinal endoscopy and small intestinal biopsy. Eleven patients (44%) among 25 patients with refractory iron deficiency anaemia, 23 patients (34.3%) among 67 patients with non-endocrinal short stature, and 6 patients (3%) among 200 patients with type I diabetes mellitus were diagnosed by jejunal biopsy as having coeliac disease. AGA (IgG) had the highest sensitivity for diagnosing CD (80.0%) followed by the TTG (72.7%) antibody, while ARA had the highest specificity (95.9%) followed by anti-EMA (94.7%). Coeliac disease is more common in Egyptian children with refractory iron deficiency anaemia, non-endocrinal short stature and type 1 diabetes than was previously thought; therefore it is mandatory to screen such patients for CD. Serological tests showed fairly good sensitivity and specificity for the diagnosis; however, intestinal biopsy remains the cornerstone for definitive diagnosis of patients with immunological reaction to gluten.


Reduced coronary reserve in response to short-term ischaemia and vasoactive drugs in ex vivo hearts from diabetic mice.
The aim of the present study was to compare the coronary flow (CF) reserve of ex vivo perfused hearts from type 2 diabetic (db/db) and non-diabetic (db/+) mice. The hearts were perfused in the Langendorff mode with Krebs-Henseleit bicarbonate buffer (37 degrees C, pH 7.4) containing 11 mmol L(-1) glucose as energy substrate. The coronary reserve was measured in response to three different interventions: (1) administration of nitroprusside (a nitric oxide donor), (2) administration of adenosine and (3) production of reactive hyperaemia by short-term ischaemia. Basal CF was approximately 15% lower in diabetic when compared with non-diabetic hearts (2.1 +/- 0.1 vs. 2.6 +/- 0.2 mL min(-1)). The maximum increase in CF rate in response to sodium nitroprusside and adenosine was significantly lower in diabetic (0.6 +/- 0.1 and 0.9 +/- 0.1 mL min(-1) respectively) than in non-diabetic hearts (1.2 +/- 0.1 and 1.4 +/- 0.1 mL min(-1) respectively). Also, there was a clear difference in the rate of return to basal CF following short-term ischaemia between diabetic and non-diabetic hearts. Thus, basal tone was restored 1-2 min after the peak hyperaemic response in non-diabetic hearts, whereas it took approximately 5 min in diabetic hearts. These results show that basal CF, as well as the CF reserve, is impaired in hearts from type 2 diabetic mice. As diabetic and non-diabetic hearts were exposed to the same (maximum) concentrations of NO or adenosine, it is suggested that the lower coronary reserve in type 2 diabetic hearts is, in part, because of a defect in the intracellular pathways mediating smooth muscle relaxation.


Isavuconazole and Liposomal Amphotericin B as Successful Combination Therapy of Refractory Invasive Candidiasis in a Liver Transplant Recipient: A Case Report and Literature Review.
Invasive fungal infections in liver transplant recipients are associated with elevated morbidity and mortality and pose a challenge to the treating physicians. Despite of lacking clinical data, the use of antifungal combination therapy is often considered to improve response rates in an immunocompromised patient population. We herein report a case of refractory invasive candidiasis in a liver transplant recipient treated successfully with a combination of isavuconazole und high-dose liposomal amphotericin B. The antimycotic combination treatment was able to clear a bloodstream infection with C. glabrata and led to regression of bilomas among tolerable side effects. The use of the above-mentioned antifungal combination therapy in a liver transplant recipient has not been reported previously. This case highlights the efficacy and safety of antifungal combination therapy in immunocompromised patients with refractory invasive candidiasis.