Conserved glutamate residues Glu-343 and Glu-519 provide mechanistic insights into cation/nucleoside cotransport by human concentrative nucleoside transporter hCNT3.
Human concentrative nucleoside transporter 3 (hCNT3) utilizes electrochemical gradients of both Na(+) and H(+) to accumulate pyrimidine and purine nucleosides within cells. We have employed radioisotope flux and electrophysiological techniques in combination with site-directed mutagenesis and heterologous expression in Xenopus oocytes to identify two conserved pore-lining glutamate residues (Glu-343 and Glu-519) with essential roles in hCNT3 Na(+)/nucleoside and H(+)/nucleoside cotransport. Mutation of Glu-343 and Glu-519 to aspartate, glutamine, and cysteine severely compromised hCNT3 transport function, and changes included altered nucleoside and cation activation kinetics (all mutants), loss or impairment of H(+) dependence (all mutants), shift in Na(+):nucleoside stoichiometry from 2:1 to 1:1 (E519C), complete loss of catalytic activity (E519Q) and, similar to the corresponding mutant in Na(+)-specific hCNT1, uncoupled Na(+) currents (E343Q). Consistent with close-proximity integration of cation/solute-binding sites within a common cation/permeant translocation pore, mutation of Glu-343 and Glu-519 also altered hCNT3 nucleoside transport selectivity. Both residues were accessible to the external medium and inhibited by p-chloromercuribenzene sulfonate when converted to cysteine.


Assessment of microbiology students' progress with an audience response system.
The development of new approaches to teaching of large lecture courses is needed. Today's classroom has a wide range of students including high-achieving motivated learners, students struggling to understand basic concepts, and learning-challenged students. Many of these students can be lost in large classes under the shadow of the high-achieving extroverted students who dominate classroom question-and-answer sessions. Measuring a student's understanding and achievement of content standards becomes difficult until an assessment has been done. To close this gap, an audience response system was introduced in an introductory Principles of Microbiology course. This technology specifically addressed the goal of individualizing instruction to the needs of the students. The evaluation of this project indicated an overall positive impact on student learning.


Mortality 10 Years After Percutaneous or Surgical Revascularization in Patients With Total Coronary Artery Occlusions.
The long-term clinical benefit after percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) in patients with total occlusions (TOs) and complex coronary artery disease has not yet been clarified. The objective of this analysis was to assess 10-year all-cause mortality in patients with TOs undergoing PCI or CABG. This is a subanalysis of patients with at least 1 TO in the SYNTAXES (Synergy Between PCI With Taxus and Cardiac Surgery Extended Survival) study, which investigated 10-year all-cause mortality in the SYNTAX (Synergy Between PCI With Taxus and Cardiac Surgery) trial, beyond its original 5-year follow-up. Patients with TOs were further stratified according to the status of TO recanalization or revascularization. Of 1,800 randomized patients to the PCI or CABG arm, 460 patients had at least 1 lesion of TO. In patients with TOs, the status of TO recanalization or revascularization was not associated with 10-year all-cause mortality, irrespective of the assigned treatment (PCI arm: 29.9% vs. 29.4%; adjusted hazard ratio [HR]: 0.992; 95% confidence interval [CI]: 0.474 to 2.075; p = 0.982; and CABG arm: 28.0% vs. 21.4%; adjusted HR: 0.656; 95% CI: 0.281 to 1.533; p = 0.330). When TOs existed in left main and/or left anterior descending artery, the status of TO recanalization or revascularization did not have an impact on the mortality (34.5% vs. 26.9%; adjusted HR: 0.896; 95% CI: 0.314 to 2.555; p = 0.837). At 10-year follow-up, the status of TO recanalization or revascularization did not affect mortality, irrespective of the assigned treatment and location of TOs. The present study might support contemporary practice among high-volume chronic TO-PCI centers where recanalization is primarily offered to patients for the management of angina refractory to medical therapy when myocardial viability is confirmed. (Synergy Between PCI With TAXUS and Cardiac Surgery: SYNTAX Extended Survival [SYNTAXES]; NCT03417050; SYNTAX Study: TAXUS Drug-Eluting Stent Versus Coronary Artery Bypass Surgery for the Treatment of Narrowed Arteries [SYNTAX]; NCT00114972).


Ocular Versus Oral Propranolol for Prevention and/or Treatment of Oxygen-Induced Retinopathy in a Rat Model.
Purpose: Propranolol, a nonselective B1/B2 adrenoceptor antagonist, promotes the regression of infantile hemangiomas likely through suppression of vascular endothelial growth factor (VEGF), which prompted its use for the prevention of retinopathy of prematurity. We tested the hypothesis that topical ocular propranolol is safe and effective for reducing the severity of oxygen-induced retinopathy (OIR) in the neonatal rat intermittent hypoxia (IH) model. Methods: At birth (P0), rat pups were randomly assigned to room air or neonatal intermittent hypoxia (IH) consisting of 50% O2 with brief episodes of hypoxia (12% O2) from P0 to P14, during which they received a single daily dose of oral propranolol (1 mg/kg/day in 50 μL in sterile normal saline) or topical ocular propranolol (0.2% in 10 μL in normal saline) from P5 to P14. Placebo-controlled littermates received 50 μL oral or 10 μL topical ocular sterile normal saline. Retinal vascular and astrocyte integrity; retinal histopathology and morphometry; and angiogenesis biomarkers were determined. Results: Topical ocular propranolol improved retinal vascular damage and preserved the astrocytic template, but did not completely prevent OIR. The beneficial effects of propranolol were associated with reduced ocular VEGF and increased endogenous soluble inhibitor, sVEGFR-1, when administered topically. Conclusions: Propranolol failed to completely prevent severe OIR, however, it prevented astrocyte degeneration resulting from neonatal IH-induced damage. We conclude that the mechanisms of propranolol's beneficial effects in neonatal IH may involve in part, astrocyte preservation.


Redetermination of Na(3)TaF(8).
The crystal structure of trisodium octafluoridotantalate, Na(3)TaF(8), has been redetermined using diffractometer data collected at 153 K, resulting in more accurate bond distances and angles than obtained from a previous structure determination based on film data. The structure is built from layers running along [101], which are formed by distorted [TaF(8)] antiprisms and [NaF(6)] rectangular bipyramids sharing edges and corners. The individual layers are separated by eight-coordinated Na ions. Two atoms in the asymmetric unit are in special positions: the Ta atom is on a twofold axis in Wyckoff position 4e and one of the Na ions lies on an inversion centre in Wyckoff site 4d.


Treatment of ankylosed deciduous molars with or without permanent successors in children and adolescents: a systematic review.
The aim of this systematic review was to provide guidelines for decision-making during orthodontic treatment planning of infra-occluded deciduous molars with or without their successors in children and adolescents. Computerized search was conducted on Medline via PubMed, and Cochrane Library. Articles published until 2020 in English language were analyzed following the Preferred Reporting Items for Systematic Reviews (PRISMA) Checklist. Observational and interventional longitudinal studies reporting the treatment of ankylosed deciduous molars with or without successor tooth in 3 to 15-year-old patients were included. In case of ankylosis with presence of successor, exfoliation took place in 77% of teeth, while extraction involved 23%. Infra-occlusion happened in 53% of teeth (worsening in 52%), alveolar bone loss in 37%, mesial tipping of first permanent molar in 5%, and over-eruption of antagonist in no cases (after exfoliation and eruption of successor). In case of ankylosis without successor, exfoliation took place in 1% of teeth, progression of infra-occlusion in 42%, progression of root resorption in 58%, development of mesial tipping of first permanent molars in 25%, while no case of antagonist over-eruption was reported. When the permanent tooth is present and the ankylosed tooth is slightly or moderately infra-occluded, observation is appropriate. In case of severe infra-occlusion or absence of successor, tooth extraction may be considered together with orthodontic space closure, transplantation, or prosthetic replacement. Alternatively, nonextraction and a prosthetic build-up may be considered.


Complications associated with 355 flexible colonoscopic procedures in dogs.
Flexible colonoscopy is commonly performed in dogs with signs of large-bowel diseases. Although considered to be a safe procedure, no reports of complications associated with colonoscopy have appeared in the veterinary literature. The purpose of this study was to describe the frequency and types of adverse events that developed during flexible colonoscopy in dogs. Medical records were reviewed from 355 scheduled colonoscopic procedures. Major complications were defined as adverse events in which the dog's life was potentially jeopardized and the complication required intensive treatment or monitoring. Major complications consisting of fatal aspiration of GoLYTELY, colonic perforation, and excessive hemorrhage after biopsy of an adenocarcinoma with rigid forceps occurred in 3 (0.85%) dogs. Minor complications associated with anesthesia or colonoscopy occurred during 3.4% of procedures. Complications were classified as minor if the adverse event required minimal treatment or monitoring, and the complication was not considered a threat to the dog's life. Vomiting of GoLYTELY occurred with the administration of 4.6% of doses in 6.5% of dogs. When administering GoLYTELY, clinicians should be prepared to rapidly remove the orogastric tube and mouth speculum if vomiting occurs to reduce the potential for aspiration. In this group of dogs undergoing flexible colonoscopy, major complications occurred infrequently and minor complications developed uncommonly. Overall, minor or major complications developed during 30 (8.5%) of 355 procedures. Mortality was rare (0.28%). Flexible colonoscopy appears to be a safe procedure in dogs with signs of large-bowel diseases.


Vascular factors in diabetes and Alzheimer's disease.
The homeostasis of neuronal cells is maintained by the cerebral circulation and blood-brain barrier. In addition to age-related physiological decline, diabetes disturbs microvascular functions through mechanisms, including activation of protein kinase C, excess production of reactive oxygen species and cellular activation of the receptor for advanced glycation endproducts (RAGE). Impaired microvasculature has been correlated with pathological changes in both vascular dementia and Alzheimer's disease. Furthermore, RAGE-mediated chronic inflammation initiates a degenerative positive feedback loop between endothelium and neuronal cells. The levels of circulating CD34+ cells, which support maintenance of the microvasculature and are decreased in diabetes, have been proposed to provide a marker of the contribution of cerebrovascular factors in patients with cognitive impairment.


Characterization of heat injury in grapes using h nuclear magnetic resonance methods : changes in transverse relaxation times.
Transverse relaxation times (T(2)) of tissue water ((1)H) in leaves and suspension cultured cells of grape hybrids (Vitis spp. cv ;Venus' and ;Veeblanc') were measured by nuclear magnetic resonance at various temperatures. The tissue water was characterized by two T(2) time constants. A sharp decrease in T(2) for the major fraction of tissue water was observed in association with heat injury, as measured by electrolyte leakage and triphenyltetrazolium chloride reduction in both leaves and suspension cultured cells. The changes in T(2) as a result of heat injury were irreversible, as indicated by a temperature dependent hysteresis of T(2). Studies using a paramagnetic probe (Mn(+2)) indicated that the plasma membrane was irreversibly damaged at the killing temperature, resulting in a loss of cell compartmentalization. Tissue water in heat-killed samples was characterized by only a single T(2).


Lithium inhibits the development of physical dependence to clonidine in mice.
Based on our previous finding that chronic lithium treatment reduced naloxone-precipitated withdrawal syndrome in morphine-treated mice, the effect of chronic lithium treatment was evaluated on the development of dependence to clonidine. Dependence was induced by injection of either morphine (50, 50 and 75 mg/kg, intraperitoneally with 3 hr interval for 3 consecutive days), or clonidine (2 mg/kg/day, intraperitoneally for 10 days). Naloxone (4 mg/kg, intraperitoneally) precipitated withdrawal signs in both morphine- and clonidine-treated mice. Yohimbine (5 mg/kg, intraperitoneally) precipitated withdrawal signs in the clonidine-treated mice, similar to morphine withdrawal signs; but failed to precipitate any significant sign in the morphine-treated mice. Coadministration of lithium was carried out by adding lithium chloride to drinking water (600 mg/l for 20 days; 10 days before the beginning of clonidine administration and 17 days before the administration of morphine to allow the lithium concentration to reach steady-state). The results indicated that chronic lithium administration significantly attenuated the withdrawal signs, precipitated either by yohimbine or naloxone, in clonidine-treated mice. As a conclusion, clonidine withdrawal signs are very similar to opioid withdrawal signs, and lithium is able to prevent the development of physical dependence to clonidine.


Amino Acid-Modified Conjugated Oligomer Self-Assembly Hydrogel for Efficient Capture and Specific Killing of Antibiotic-Resistant Bacteria.
Bacterial infection is one of main causes that threaten global human health. Especially, antibiotic-resistant bacteria like methicillin-resistant Staphylococcus aureus (MRSA) lead to high mortality rate and more expensive treatment cost. Here, a novel amino-acid-modified conjugated oligomer OTE-d-Phe was synthesized by modifying the side chain of conjugated oligo(thiophene ethynylene) with d-phenylalanine. By mixing 9-fluorenylmethyloxycarbonyl-l-phenylalanin (Fmoc-l-Phe) with OTE-d-Phe, a new and biocompatible low-molecular weight hydrogel (HG-2) was prepared through self-assembly. In solution, HG-2 can effectively capture bacteria spontaneously, such as Escherichia coli and MRSA. Most importantly, the hydrogel has specific and strong antibacterial activity against MRSA over methicillin-susceptible S. aureus, Staphylococcus epidermidis, and E. coli. Interestingly, when the hydrogel was put on a model surface, a piece of cloth, it also is able to selectively kill MRSA with low cell cytotoxicity. The antibacterial mechanism was investigated, and it demonstrated that the HG-2 interacts with and physically breaks the cell wall and membrane, which leads to MRSA death. Therefore, this new conjugated oligomer-based hydrogel provides promising applications in disinfection and therapy of MRSA in hospital and in community.


Crosstalk Between Pheromone Signaling and NADPH Oxidase Complexes Coordinates Fungal Developmental Processes.
Sexual and asexual development in filamentous ascomycetes is controlled by components of conserved signaling pathways. Here, we investigated the development of mutant strains lacking genes for kinases MAK2, MEK2, and MIK2, as well as the scaffold protein HAM5 of the pheromone response (PR) pathway. All had a defect in fruiting body development and hyphal fusion. Another phenotype was a defect in melanin-dependent ascospore germination. However, this deficiency was observed only in kinase deletion mutants, but not in strains lacking HAM5. Notably, the same developmental phenotypes were previously described for nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 1 (NOX1) mutants, but the germination defect was only seen in NOX2 mutants. These data suggest a molecular link between the pheromone signaling pathway and both NOX complexes. Using data from yeast two-hybrid (Y2H) analysis, we found that the scaffolding protein HAM5 interacts with NOR1, the regulator of NOX1 and NOX2 complexes. This interaction was further confirmed using differently fluorescent-labeled proteins to demonstrate that NOR1 and HAM5 co-localize at cytoplasmic spots and tips of mature hyphae. This observation was supported by phenotypic characterization of single and double mutants. The oxidative stress response and the initiation of fruiting bodies were similar in Δham5Δnor1 and Δham5, but distinctly reduced in Δnor1, indicating that the double deletion leads to a partial suppression of the Δnor1 phenotype. We conclude that the PR and NOX1 complexes are connected by direct interaction between HAM5 and NOR1. In contrast, PR kinases are linked to the NOX2 complex without participation of HAM5.


Regeneration of Cartilage in Human Knee Osteoarthritis with Autologous Adipose Tissue-Derived Stem Cells and Autologous Extracellular Matrix.
This clinical case series demonstrates that percutaneous injections of autologous adipose tissue-derived stem cells (ADSCs) and homogenized extracellular matrix (ECM) in the form of adipose stromal vascular fraction (SVF), along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, could regenerate cartilage-like tissue in human knee osteoarthritis (OA) patients. Autologous lipoaspirates were obtained from adipose tissue of the abdominal origin. Afterward, the lipoaspirates were minced to homogenize the ECM. These homogenized lipoaspirates were then mixed with collagenase and incubated. The resulting mixture of ADSCs and ECM in the form of SVF was injected, along with HA and PRP activated by calcium chloride, into knees of three Korean patients with OA. The same affected knees were reinjected weekly with additional PRP activated by calcium chloride for 3 weeks. Pretreatment and post-treatment magnetic resonance imaging (MRI) data, functional rating index, range of motion (ROM), and pain score data were then analyzed. All patients' MRI data showed cartilage-like tissue regeneration. Along with MRI evidence, the measured physical therapy outcomes in terms of ROM, subjective pain, and functional status were all improved. This study demonstrates that percutaneous injection of ADSCs with ECM contained in autologous adipose SVF, in conjunction with HA and PRP activated by calcium chloride, is a safe and potentially effective minimally invasive therapy for OA of human knees.


Glucagon-like peptide-1 receptor activation antagonizes voltage-dependent repolarizing K(+) currents in beta-cells: a possible glucose-dependent insulinotropic mechanism.
Glucagon-like peptide-1 (GLP-1) acts through its G-protein-coupled receptor to enhance glucose-stimulated insulin secretion from pancreatic beta-cells. This is believed to result from modulation of at least two ion channels: ATP-sensitive K(+) (K(ATP)) channels and voltage-dependent Ca(2+) channels. Here, we report that GLP-1 receptor signaling also regulates the activity of beta-cell voltage-dependent K(+) (K(V)) channels, themselves potent glucose-dependent regulators of insulin secretion. GLP-1 receptor activation with exendin 4 (10(-8) mol/l) in rat beta-cells antagonized K(V) currents by 43.3 +/- 6.3%, whereas the GLP-1 receptor antagonist exendin 9-39 had no effect. The effect of GLP-1 receptor activation on K(V) currents could be replicated (current reduction of 55.7 +/- 6.0%) by G-protein activation with GMP-PNP (10 nmol/l). The cAMP pathway antagonist Rp-cAMPS (100 micro mol/l) prevented current inhibition by exendin 4, implicating cAMP signaling in GLP-1 receptor modulation of beta-cell K(V) currents. Finally, exendin 4 (10(-8) mol/l) increased the amplitude (130 +/- 5.7%) and duration (285 +/- 15.9%) of the beta-cell depolarization response to current injection, independent of any effect on K(ATP) or Ca(2+) channels. The present results demonstrate that GLP-1 receptor signaling can antagonize beta-cell repolarization by reducing voltage-dependent K(+) currents, an effect likely to contribute to GLP-1's glucose-dependent insulinotropic effect.


Identification of an endothelin-converting enzyme-2-specific fluorigenic substrate and development of an in vitro and ex vivo enzymatic assay.
Endothelin-converting enzyme-2 (ECE-2) is a membrane-bound zinc-dependent metalloprotease that shares a high degree of sequence homology with ECE-1, but displays an acidic pH optimum characteristic of maturing enzymes acting late in the secretory pathway. Although ECE-2, like ECE-1, can cleave the big endothelin intermediate to produce the vasoconstrictive endothelin peptide, its true physiological function remains to be elucidated, a task that is hampered by the lack of specific tools to study and discriminate ECE-2 from ECE-1, i.e. specific substrates and/or specific inhibitors. To fill this gap, we searched for novel ECE-specific peptide substrates. To this end, peptides derived from the big endothelin intermediate were tested using ECE-1 and ECE-2, leading to the identification of an ECE-1-specific substrate. Moreover, screening of our proprietary fluorigenic peptide Fluofast® libraries using ECE-1 and ECE-2 allowed the identification of Ac-SKG-Pya-F-W-Nop-GGK-NH(2) (PL405), as a specific and high affinity ECE-2 substrate. Indeed, ECE-2 cleaved PL405 at the Pya-F amide bond with a specificity constant (k(cat)/K(m)) of 8.1 ± 0.9 × 10(3) M(-1) s(-1). Using this novel substrate, we also characterized the first potent (K(i) = 7.7 ± 0.3 nM) and relatively selective ECE-2 inhibitor and developed a quantitative fluorigenic ECE-2 assay. The assay was used to study the ex vivo ECE-2 activity in wild type and ECE-2 knock-out tissues and was found to truly reflect ECE-2 expression patterns. The PL405 assay is thus the first tool to study ECE-2 inhibition using high throughput screening or for ex vivo ECE-2 quantification.


Purine metabolism and the microaerophily of Helicobacter pylori.
The requirements for purine nucleotide synthesis, the effects of purine analogues, and the metabolism of adenine in the bacterium Helicobacter pylori were investigated employing cell culture techniques and one-dimensional NMR spectroscopy. Bacterial cells grew and proliferated in media lacking preformed purines, indicating that H. pylori can synthesize purine nucleotides de novo to meet its requirements. Blocking of this pathway in the absence of sufficient preformed purines for salvage nucleotide synthesis led to cell death. Analogues of purine nucleobases and nucleosides taken up by the cells were cytotoxic, suggesting that salvage routes could be exploited for therapy. Adenine or hypoxanthine were able to substitute for catalase in supporting cell growth and proliferation, suggesting a role for these bases in maintaining the microaerophilic conditions essentially required by the bacterium.


Measurement of blood flow in rat eyes by hydrogen clearance.
Intraocular measurements of blood flow have been made in the rat eye using hydrogen-clearance polarography. Hydrogen was delivered by a bolus injection of hydrogen-saturated saline via a cannula in the lingual artery of the anesthetized rat. An intraocularly placed hydrogen-sensitive microelectrode monitored the arrival and clearance of hydrogen preretinally adjacent to retinal arteries, veins, and intervascular areas. The effect of stepwise penetration through the retina and into the choroid was also determined, and a map of hydrogen distribution in the retina during hydrogen delivery and clearance was produced. Intraocular measurements, which have the advantage that the electrode location can be clearly visualized, together with computer control of the bolus injection and data collection, have allowed highly repeatable measurements of local blood flow to be made. The mean blood flow in retinal tissue was 373 +/- 51 ml.min-1.100 g-1 (values are means +/- SD, n = 32). The correlation between the nature of the hydrogen clearance curve and the location of the electrode helps explain data from other studies in which the position of the electrode with respect to the vasculature was unknown.


Effects of abstinence on cigarette smoking among outpatients with schizophrenia.
The authors of this study examined the effects of brief smoking abstinence on smoking among 6 individuals with schizophrenia or schizoaffective disorder. Before 6 of 12 experimental sessions, participants were required to provide breath carbon monoxide (CO) samples indicative of smoking abstinence; before the remaining sessions, participants provided CO samples indicating no abstinence. During sessions, participants obtained smoking opportunities (2 puffs/opportunity) under either fixed ratio-1 or progressive ratio (PR) schedules of reinforcement. Abstinence increased smoking under both schedules and increased breakpoint for smoking under the PR schedule. These data offer further evidence that smoking by individuals with schizophrenia is orderly, operant behavior that is modulated, at least in part, by variables that also affect smoking in people without major mental illness.


Computed Tomography-based Subclassification of Chronic Obstructive Pulmonary Disease.
Computed tomography (CT) is an obvious modality for subclassification of COPD. Traditionally, the pulmonary involvement of chronic obstructive pulmonary disease (COPD) in smokers is understood as a combination of deleterious effects of smoking on small airways (chronic bronchitis and small airways disease) and distal to the airways with destruction and loss of lung parenchyma (emphysema). However, segmentation of airways is still experimental; with contemporary high-resolution CT (HRCT) we can just see the "entrance" of small airways, and until now changes in airway morphology that have been observed in COPD are subtle. Furthermore, recent results indicate that emphysema may also be the essential pathophysiologic mechanism behind the airflow limitation of COPD. The definition of COPD excludes bronchiectasis as a symptomatic subtype of COPD, and CT findings in chronic bronchitis and exacerbations of COPD are rather unspecific. This leaves emphysema as the most obvious candidate for subclassification of COPD. Both chest radiologists and pulmonary physicians are quite familiar with the appearance of various patterns of emphysema on HRCT, such as centrilobular, panlobular, and paraseptal emphysema. However, it has not yet been possible to develop operational definitions of these patterns that can be used by computer software to automatically classify CT scans into distinct patterns. In conclusion, even though various emphysema patterns can be recognized visually, CT has not yet demonstrated a great potential for automated subclassification of COPD, and it is an open question whether it will ever be possible to achieve success equivalent to that obtained by HRCT in the area of interstitial lung diseases.


Discrepant expression of membrane and soluble isoforms of Fas (CD95/APO-1) in adult T-cell leukaemia: soluble Fas isoform is an independent risk factor for prognosis.
The Fas signalling system probably plays a critical role in the natural and chemotherapeutic cell death machinery, suggesting that aberrant Fas expression is involved in growth control of tumours. The membrane isoform (mFas) is a 45 kD cell surface protein containing a single transmembrane region, and induces apoptosis in normal or tumour cells, whereas the soluble isoform (sFas) lacks the transmembrane domain due to alternative splicing of the transcript and is thought to block Fas-mediated apoptosis. To clarify the clinical roles of expression of these two Fas isoforms in adult T-cell leukaemia (ATL), we investigated the levels of the Fas isoforms in 81 patients with ATL. The expression patterns of the Fas isoforms were heterogenous, and there was no significant correlation between mFas and sFas levels: 10/81 cases were negative for mFas and had high serum sFas levels, whereas the remaining 71 cases were positive for mFas and had various levels of expression of the two Fas isoforms. Irrespective of the status of mFas expression in leukaemic cells, the mRNAs encoding these isoforms were always detectable, indicating the potential for protein translation. Although mFas expressed on freshly isolated ATL cells could iduce apoptosis in vitro, positive versus negative mFas status was not associated with any clinical aspects of ATL, whereas the sFas level was strongly correlated with clinical parameters such as serum LDH activity, tumour burden, serum soluble IL-2R level, hypercalcaemia and prognosis. These results suggest that the ratio of Fas isoforms varies, and high expression of the sFas protein and message reflects the malignant behaviour of ATL and is an independent risk factor for the prognosis.


[Hysterosonography].
Hysterosonography, which consists in the injection of some cc of saline in the uterine cavity during sonography, allows an exploration of the uterine cavity and of the endometrium. Accuracy of hysterosonography is similar to that of office hysteroscopy. Hysterosonography distinguishes normal cavities from pathologic ones, endometrial atrophy from mucosal anomalies, polyps from myomas. Conversely, endometrial biopsy is still necessary for diagnosis of hyperplasia or cancer. Hysterosonography can also be proposed for first trimester bleedings, trophoblast retention or ectopic pregnancy. In the near future it could be applied to therapeutic goals such as sonographic section of polyps.


Correlation of cutaneous lesions with clinical radiological and urodynamic findings in the prognosis of underlying spinal dysraphism disorders.
Spinal dysraphism is characterized by a lack of fusion of the vertebral arches that occurs in the absence of spinous processes with variable amounts of lamina. Here, we retrospectively present the importance of cutaneous lesions and their correlation with clinical presentation, radiological examination and urodynamic assessment. We retrospectively reviewed 612 (6.12%) cases with skin lesions from 10,000 consecutive live-born children seen at two institutions between January 1998 and March 2005. We divided all children into a control group and three groups based on clinical assessment, radiological examination and urodynamic evaluation results. Neurological deficits were identified in 113 (18.46%) children, while spinal dysraphism disorders, tethered cord syndrome and associated orthopedic malformations were found in 171 (27.94%), 119 (19.45%) and 28 (4.57%) patients, respectively. The incidence of tethered cord with lumbosacral dimple lesions over the sacrum (32/119, 26.65% of patients) was 3.5-fold higher than that of lesions found over the coccyx (9/119, 7.5% of patients). Uroneurological symptoms were found in 207/612 (33.82%) children. Urodynamic assessment revealed decreased bladder capacity in 10% of patients, detrusor hyperflexia during filling in 47% and a low-compliance detrusor in 71%. Discordance between ultra- sonography and MRI was found in 16.58% of patients. Spinal cord untethering was performed for 109 patients. Nearly all children with resolution were at the end of their follow-up period (24 months). Retethering occurred in 21 (19.26%) patients, and a second untethering surgery was performed in 12 patients. When spina bifida was associated with lumbar skin lesions, there may have been an increased incidence of tethered cord and other spinal cord disorders. MRI scans are more reliable and give an exact diagnosis of tethered cord. Neurological and uroneurological instability are ultimately a clinical diagnosis, and there is controversy about their indications for surgery. However, the correlation between urodynamic assessment and cutaneous lesions with a tethered cord found by MRI examination allow for an early diagnosis and the possibility of prompt treatment.


Utilization of Isometric Strength Training and Interval Training With a Patient With Cancer in the Acute Care Setting.
The onset of cancer and subsequent treatments often result in deficits in physical function and quality of life (QoL). Available research has demonstrated that an individualized exercise program has the ability to reduce fatigue, optimize physical function, and improve QoL. However, the exercise program is often lacking appropriate intensity and volume resulting in negligent improvement or even further reduction in function. Thus, the purpose of this case report is to demonstrate the application of isometric strength training (IST) and high-intensity interval training (HIIT) in the acute care setting with an individual with a history of malignant mesothelioma. The patient demonstrated improvement in functional mobility evidenced by improvement in Activity Measure for Post-Acute Care (AM-PAC) score through the hospitalization along with increased ambulation distance. No adverse events occurred during any physical therapy (PT) visits while utilizing HIIT or IST.


Different effects of indomethacin on healing of shaft and metaphyseal fractures.
NSAIDs are commonly used in the clinic, and there is a general perception that this does not influence healing in common types of human fractures. Still, NSAIDs impair fracture healing dramatically in animal models. These models mainly pertain to fractures of cortical bone in shafts, whereas patients more often have corticocancellous fractures in metaphyses. We therefore tested the hypothesis that the effect of an NSAID is different in shaft healing and metaphyseal healing. 26 mice were given an osteotomy of their left femur with an intramedullary nail. 13 received injections of indomethacin, 1 mg/kg twice daily. After 17 days of healing, the femurs were analyzed with 3-point bending and microCT. 24 other mice had holes drilled in both proximal tibias, to mimic a stable metaphyseal injury. A screw was inserted in the right tibial hole only. After 7 days of indomethacin injections or control injections, screw fixation was measured with mechanical pull-out testing and the side without a screw was analyzed with microCT. In the shaft model, indomethacin led to a 35% decrease in force at failure (95% CI: 14-54). Callus size was reduced to a similar degree, as seen by microCT. Metaphyseal healing was less affected by indomethacin, as no effect on pull-out force could be seen (95% CI: -27 to 17) and there was only a small drop in new bone volume inside the drill hole. The difference in the relative effect of indomethacin between the 2 models was statistically significant (p = 0.006). Indomethacin had a minimal effect on stable metaphyseal fractures, but greatly impaired healing of unstable shaft fractures. This could explain some of the differences found between animal models and clinical experience.


Crystallization of the C-terminal domain of rabbit serum hemopexin.
The C-terminal domain of rabbit serum hemopexin, comprising residues 215 to 435, has been crystallized following removal of the attached carbohydrate using the endoglycosidase Endo F. The crystals, grown by vapour diffusion from solutions containing polyethylene glycol 1500, are orthorhombic, with cell dimensions a = 41.0 A, b = 64.2 A, c = 85.2 A, space group P2(1)2(1)2(1), and one molecule in the asymmetric unit. The crystals diffract to 2.4 A resolution and are suitable for X-ray structure analysis.


Menstrual Cycle Patterns and Irregularities in Hired Latinx Child Farmworkers.
This study identifies the menstrual cycle irregularities of Latinx child and adolescent farmworkers. Child and adolescent farmworkers aged 13 to 20 years completed questionnaires about menstrual cycle patterns in 2019, and wore silicone passive collection wristbands for pesticide detection in 2018. Menstrual cycle irregularities were determined from the American College of Obstetricians and Gynecologists committee opinion. Half of participants experienced any menstrual cycle irregularity; the most frequent irregularities were cycle length (38.6%) and having gone 90 days or more without a menstrual period (20.4%). Pesticides were detected in 92.9% of the wristbands; most participants were exposed to an endocrine disrupting chemical (EDC) pesticide. Half of Latinx children and adolescents hired farmworkers experience irregular menstrual cycles, and most are exposed to EDCs. Inclusion of occupational and menstrual histories in child and adolescent medical visits is critical.


Attenuation of sodium arsenite-induced cardiotoxicity and neurotoxicity with the antioxidant, anti-inflammatory, and antiapoptotic effects of hesperidin.
In the scope of the study, the protective effect of hesperidin (HES), a flavanone glycoside, was investigated against sodium arsenite (NaAsO2, SA) induced heart and brain toxicity. For this purpose, 35 Sprague-Dawley male rats were divided into 5 different groups, 7 in each group. Physiological saline was given to the first group. Dose of 200 mg/kg of HES to the second group, 10 mg/kg dose of SA to the 3rd group, 100 mg/kg HES and 10 mg/kg SA to the 4th group, 200 mg/kg HES, and 10 mg/kg SA to the 5th group were given orally for 15 days. At the end of the study, biochemical, histopathological, and immunohistochemical examinations were performed on the heart and brain tissues of the rats. According to the results, SA increased malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels and decreased glutathione (reduced, GSH) level and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities in both tissues. Also, it increased cardiac lactate dehydrogenase (LDH) and creatine kinase isoenzyme-MB (CK-MB) activities and cardiac troponin-I level (cTn-I), cerebral acetylcholine esterase activity, nuclear factor kappa-B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-one beta (IL-1β), and cysteine aspartate-specific protease-3 (caspase-3) levels. In addition, as a result of histopathological examination, it was determined that SA damaged tissue architecture, and as a result of immunohistochemical examination, it increased cardiac Bcl-2-associated X protein (Bax) and cerebral glial fibrillary acidic protein (GFAP) expression. The results have also shown that HES co-treatment has an antioxidant, anti-inflammatory, antiapoptotic effect on SA-induced toxicity and aids to protect tissue architecture by showing a regulatory effect on all values. Consequently, it was determined that HES co-treatment had a protective effect on SA-induced heart and brain toxicity in rats.


Elevated serum neurotensin and CRH levels in children with autistic spectrum disorders and tail-chasing Bull Terriers with a phenotype similar to autism.
Autism spectrum disorders (ASD) are neurodevelopmental disorders characterized by defects in communication and social interactions, as well as stereotypic behaviors. Symptoms typically worsen with anxiety and stress. ASD occur in early childhood, often present with regression and have a prevalence of 1 out of 68 children. The lack of distinct pathogenesis or any objective biomarkers or reliable animal models hampers our understanding and treatment of ASD. Neurotensin (NT) and corticotropin-releasing hormone (CRH) are secreted under stress in various tissues, and have proinflammatory actions. We had previously shown that NT augments the ability of CRH to increase mast cell (MC)-dependent skin vascular permeability in rodents. CRH also induced NT receptor gene and protein expression in MCs, which have been implicated in ASD. Here we report that serum of ASD children (4-10 years old) has significantly higher NT and CRH levels as compared with normotypic controls. Moreover, there is a statistically significant correlation between the number of children with gastrointestinal symptoms and high serum NT levels. In Bull Terriers that exhibit a behavioral phenotype similar to the clinical presentation of ASD, NT and CRH levels are also significantly elevated, as compared with unaffected dogs of the same breed. Further investigation of serum NT and CRH, as well as characterization of this putative canine breed could provide useful insights into the pathogenesis, diagnosis and treatment of ASD.


Effect of Gloriosa superba linn (EEGS) on mPT and monosodium glutamate-induced proliferative disorder using rat model.
Hyperplasia, Tumors and cancers are various forms of proliferative disorders affecting humans. Surgery is the main treatment approach while other options are also associated with adverse effects. There is therefore a need for the development of better alternative therapy that is cost effective and readily available with little or no adverse effect. Some bioactive agents in medicinal plants exhibit their anti-proliferative potential by induction of mitochondrial permeability transition pore (mPT) opening. Gloriosa superba, a medicinal plant, is folklorically used in the treatment of tumors and cancers. This study therefore aimed at investigating the effect of ethanol leaf extract of Gloriosa superba (EEGS) on mPT and monosodium glutamate-induced proliferative disorder in some specific tissues using rat model. Isolated rat liver mitochondria were exposed to different concentrations (10, 30, 50, 70 and 90 μg/ml) of EEGS. The mPT pore opening, cytochrome c release, mitochondrial ATPase activity and lipid peroxidation were assessed spectrophotometrically. Caspases 9 and 3 activities were carried out using ELISA technique. Histological assessment of the liver, prostate and uterus of normal and monosodium glutamate (MSG)-treated rats were carried out. The results showed significant induction of mPT pore opening, release of cytochrome c, enhancement of mitochondrial ATPase activity, inhibition of lipid peroxidation and activation of caspases 9 and 3 activities by EEGS. The histological assessment revealed the presence of MSG-induced hepato-cellular damage, benign prostate hyperplasia and uterine hyperplasia which were ameliorated by EEGS co-administration. These findings suggest that EEGS contains putative agents that can induce apoptosis via induction of mPT pore opening and as well protect against MSG-induced hepato-cellular damage and proliferative disorder in prostate and uterus.


Novel Psychrophiles and Exopolymers from Permafrost Thaw Lake Sediments.
Thermokarst lakes are one of the most abundant types of microbial ecosystems in the circumpolar North. These shallow basins are formed by the thawing and collapse of ice-rich permafrost, with subsequent filling by snow and ice melt. Until now, permafrost thaw lakes have received little attention for isolation of microorganisms by culture-based analysis. The discovery of novel psychrophiles and their biomolecules makes these extreme environments suitable sources for the isolation of new strains, including for potential biotechnological applications. In this study, samples of bottom sediments were collected from three permafrost thaw lakes in subarctic Québec, Canada. Their diverse microbial communities were characterized by 16S rRNA gene amplicon analysis, and subsamples were cultured for the isolation of bacterial strains. Phenotypic and genetic characterization of the isolates revealed affinities to the genera Pseudomonas, Paenibacillus, Acinetobacter,Staphylococcus and Sphingomonas. The isolates were then evaluated for their production of extracellular enzymes and exopolymers. Enzymes of potential biotechnological interest included α and β-glucosidase, α and β-maltosidase, β-xylosidase and cellobiohydrolase. One isolate, Pseudomonas extremaustralis strain 2ASCA, also showed the capability to produce, in the loosely bound cell fraction, a levan-type polysaccharide with a yield of 613 mg/L of culture, suggesting its suitability as a candidate for eco-sustainable alternatives to commercial polymers.


Co-occurrence of emphysematous cystitis and emphysematous myositis in type 2 diabetes.
Emphysematous cystitis (EC) and emphysematous myositis (EM) are rare disorders and concurrent occurrence of both in a patient with type 2 diabetes has not been reported previously. We report a patient who presented with pneumaturia and later with pain in thigh and diagnosed of both concurrently.


Cloning and expression of cDNA encoding human lysosomal acid lipase/cholesteryl ester hydrolase. Similarities to gastric and lingual lipases.
Molecular cloning of a full-length cDNA for human lysosomal acid lipase/cholesteryl ester hydrolase (EC 126.96.36.199) reveals that it is structurally related to previously described enteric acid lipases, but lacks significant homology with any characterized neutral lipases. The lysosomal enzyme catalyzes the deacylation of triacylglyceryl and cholesteryl ester core lipids of endocytosed low density lipoproteins; this activity is deficient in patients with Wolman disease and cholesteryl ester storage disease. Its amino acid sequence, as deduced from the 2.6-kilobase cDNA nucleotide sequence, is 58 and 57% identical to those of human gastric lipase and rat lingual lipase, respectively, both of which are involved in the preduodenal breakdown of ingested triglycerides. Notable differences in the primary structure of the lysosomal lipase that may account for discrete catalytic and transport properties include the presence of 3 new cysteine residues, in addition to the 3 that are conserved in this lipase gene family, and of two additional potential N-linked glycosylation sites. Transfection of the cDNA into Cos-1 cells resulted in the expression of acid lipase activity with the substrate range of the native enzyme at a level that was greater than 40 times the endogenous activity.


Evolving Status of Clinical Immunotherapy with Oncolytic Adenovirus.
Cancer immunotherapy targeting immune checkpoint inhibitors shows efficacy in several human cancers, but "cold tumors" that lack immune cells are typically unresponsive. Among the potential therapeutic approaches that could "heat" or promote lymphocyte infiltration of cold tumors, oncolytic viruses have attracted interest for their lytic and immunogenic mechanisms of action. In this article, we review the use of oncolytic adenoviruses in cancer immunotherapy, with a particular focus on preclinical and clinical data of oncolytic adenovirus-triggered immune responses against tumor antigens. We also discuss parameters to consider in clinical trial design and the combination of oncolytic adenoviruses with conventional treatments or other immunotherapies.


Infectious endophthalmitis in adult eyes receiving Boston type I keratoprosthesis.
To report the clinical characteristics of infectious endophthalmitis after Boston type I keratoprosthesis (K-Pro) implantation. Retrospective study. One hundred forty-one adult eyes receiving a K-Pro at a single institution from May 2004 through July 2008. A retrospective chart review was performed of all adult eyes receiving a K-Pro at the University of Rochester from May 2004 through July 2008. Those patients identified as having been treated for exogenous bacterial endophthalmitis were reviewed for demographic data, indication for K-Pro, bandage contact lens use, prophylactic antibiotic use, timing and clinical presentation of endophthalmitis, gram stain and culture results of intraocular fluid, timing and presentation of any subsequent episodes of endophthalmitis (recurrent endophthalmitis), and preoperative and postoperative visual acuity through August 2010. Incidence of endophthalmitis, time to occurrence, recurrence rates, visual outcomes, and risk factors associated with K-Pro endophthalmitis. Ten (7.1%) of 141 eyes of 130 adult patients were diagnosed and treated for bacterial endophthalmitis. Average time to endophthalmitis developing after K-Pro was 9.8 months (standard deviation [SD], 6.2 months; range, 2-25 months). Coagulase-negative staphylococci were identified in 7 eyes. In 7 of the 10 eyes, recurrent endophthalmitis developed that occurred at a mean of 4 months (SD, 3.9 months; range, 1-13 months) after resolution of the initial episode. At each episode of endophthalmitis, no eye was receiving vancomycin ophthalmic drops and most eyes were receiving only fluoroquinolone ophthalmic drops for prophylaxis. Infectious endophthalmitis after K-Pro implantation has a higher incidence, delayed onset, and high risk for recurrence compared with postoperative endophthalmitis associated with more common intraocular procedures such as cataract surgery. The concurrent use of topical vancomycin is recommended because it seems to be important in reducing the incidence and recurrence of endophthalmitis and because fluoroquinolone ophthalmic drops do not seem to be sufficient prophylaxis in these eyes.


[Nuclear medicine today].
This paper deals with those nuclear medicine investigations, which, in spite of development of other diagnostic methods and imaging procedures, remain important and in some cases, exclusive diagnostic tool in differential diagnostics of certain disorders and diseases. Examination of myocardial perfusion with radiotracers is the only non-invasive imaging method. Typically two sets of images are acquired. The first one reflecting myocardial perfusion at peak stress (exercise) and the second set reflecting perfusion at rest. Fixed defects in the scintigrams show infarcted myocardiac regions. Thallium-201 exercise-rest perfusion imaging has a strong prognostic value for coronary artery disease. Lung perfusion-ventilation scanning is an essentially non-invasive method of evaluating pulmonary embolism. Normal lung scan excludes clinically significant pulmonary embolism and patients with normal or very low probability perfusion scintigrams do not require therapy for pulmonary embolism. Radioisotopes play an important role in the diagnosis of malignant bone lesions. Bone scanning is commonly used to detect metastases from malignant tumours, because it is more sensitive than x-rays. In benign bone diseases, scintigraphy is used to evaluate sports injuries, avascular necrosis and infection of joint prostheses. Methods of nuclear medicine are useful for evaluating renal outflow obstructions, cortical and renovascular dysfunction. Normal finding of dynamic renography excludes an obstructive disease, whereas loss of radionuclide elimination after stimulation with furosemide is characteristic for it. In patients with suspected renovascular hypertension dynamic renography combined with captopril is a useful diagnostic test. In patients with fever of unknown origin imaging with labelled autologous leukocytes may be used to establish the location in gastrointestinal, neurological, psychiatric and other disorders. Radionuclide investigations are helpful in diagnostics of some gastrointestinal disorders, such as diagnosis of malabsorption of vitamin B-12, malabsorption of bile acids or diagnosis of gastrointestinal bleeding caused by ectopic gastric mucosa in persisting Meckel's diverticulum. Cerebral blood flow studies and positron emission tomography are widely used in diagnostics of various neurological and psychiatric diseases. Thyroid scintigraphy is commonly used to assess a thyroid nodule. Radioisotopes are used to localize primary malignant tumors in oncology, and also they are used in therapy of certain disorders. Nuclear medicine must remain an integral part of health care. Dental of nuclear medicine services, from professional point of view, is therefore unjustified.


Nasopharyngeal carcinoma and lymphoinfiltration.
Originally referred to as 'lymphoepithelioma', undifferentiated and poorly differentiated nasopharyngeal carcinoma (NPC) tissues showed intense lymphoinfiltration. In a study of cryosections from 15 NPC tissues, we found that infiltrating lymphoid elements were comprised predominantly of lymphocytes, but plasma cells, follicular dendritic cells, and eosinophils were also commonly seen. Subpopulations of lymphocytes having the same phenotypes tend to aggregate, forming clusters or secondary follicles in stromatous tissues. The tumor areas were mainly infiltrated by T cells. Tumor cells and/or apparently normal epithelium in the paratumorous areas frequently expressed CD21, CD23, CD40 and a B lymphocytes carcinoma cross-reacting antigen (BLCa), all of which are involved in B cell activation and proliferation. CD21 and BLCa were strongly expressed near the surface of both squamous and columnar epithelium by those epithelial cells which are at advanced stage of differentiation, while CD40 was expressed by epithelial cells at earlier stages of differentiation located at or near the basement membrane. CD23 was mainly expressed by columnar cells and basal cells underlying squamous epithelium, but not, or weakly so, by flattened squamous cells or reserve cells underlying columnar epithelium. The large majority of tumor cells expressed CD40 and BLCa. A substantial proportion of them also expressed CD23, but the tumor cells were not reactive for CD21. Despite eosinophilic infiltration, IL-6 was not detected in tumor tissues. IL-1 was, however, detected in abundance in the cytoplasm of follicular dendritic-like cells and in the intercellular spaces in tumor areas and surrounding stromatous tissues. The immunobiology of NPC is discussed in the light of these observations.


Incidence and Risk Factors of Postoperative Bleeding in Patients Undergoing Total Thyroidectomy.
Purpose: Our goal was to analyze postoperative bleeding in patients undergoing total thyroidectomy and to explore the possible risk factors. Materials and Methods: Patients undergoing total thyroidectomy were retrospectively enrolled, and the main study outcomes were postoperative bleeding and 30-day mortality. Univariate and multivariate analyses were used to determine the independent risk factors for postoperative bleeding. Results: A total of 31,706 patients were enrolled for analysis during January 2010 and December 2018 from the Affiliated First Hospital of Zhengzhou University. Benign and malignant disease was reported in 4,521 and 27,185 patients, respectively. Postoperative bleeding occurred in 48 patients with benign disease and in 263 patients with malignant disease. There was one bleeding site in 243 patients. The branch of the superior thyroid artery was the most common arterial bleeding site, occurring in 124 patients, and the anterior jugular vein was the most common venous bleeding site, occurring in 85 patients. Multivariable analysis confirmed that hypertension, diabetes, BMI, and disease pathology were independent factors affecting postoperative bleeding in patients with benign disease and that hypertension, diabetes, BMI, operation time, tumor stage, and tracheotomy were independent factors affecting postoperative bleeding in patients with malignant disease. In patients with postoperative bleeding, there were 5 deaths; in patients without postoperative bleeding, there were 42 deaths, and the difference was significant (p < 0.001). Conclusions: Compared with malignant disease patients, benign disease patients have a similar postoperative bleeding rate. A previous history of chemotherapy or radiotherapy has no significant effect on postoperative bleeding.


Social Isolation in Adolescence Disrupts Cortical Development and Goal-Dependent Decision-Making in Adulthood, Despite Social Reintegration.
The social environment influences neurodevelopment. Investigations using rodents to study this phenomenon commonly isolate subjects, then assess neurobehavioral consequences while animals are still isolated. This approach precludes one from dissociating the effects of on-going versus prior isolation, hindering our complete understanding of the consequences of social experience during particular developmental periods. Here, we socially isolated adolescent mice from postnatal day (P)31 to P60, then re-housed them into social groups. We tested their ability to select actions based on expected outcomes using multiple reinforcer devaluation and instrumental contingency degradation techniques. Social isolation in adolescence (but not adulthood) weakened instrumental response updating, causing mice to defer to habit-like behaviors. Habit biases were associated with glucocorticoid insufficiency in adolescence, oligodendrocyte marker loss throughout cortico-striatal regions, and dendritic spine and synaptic marker excess in the adult orbitofrontal cortex (OFC). Artificial, chemogenetic stimulation of the ventrolateral OFC in typical, healthy mice recapitulated response biases following isolation, causing habit-like behaviors. Meanwhile, correcting dendritic architecture by inhibiting the cytoskeletal regulatory protein ROCK remedied instrumental response updating defects in socially isolated mice. Our findings suggest that adolescence is a critical period during which social experience optimizes one's ability to seek and attain goals later in life. Age-typical dendritic spine elimination appears to be an essential factor, and in its absence, organisms may defer to habit-based behaviors.


Extracorporeal membrane oxygenation for pneumonitis after a Glenn palliation.
A five-month old male with a single ventricle palliated with a bidirectional cavopulmonary anastomosis developed severe respiratory insufficiency from respiratory syncytial virus (RSV) pneumonitis. He was successfully rescued with extracorporeal membrane oxygenation (ECMO) therapy and recovered with minimal morbidity.


Neoadjuvant chemotherapy in invasive bladder cancer.
The role of neoadjuvant chemotherapy in muscle-invasive bladder cancer has been clarified by recent randomized studies and meta-analyses, which all showed that cisplatin-based, combination chemotherapy offers a significant survival advantage. Preoperative chemotherapy results in downstaging in a significant percentage of patients, which is an independent factor of favorable prognosis. Nevertheless, the optimal sequence of perioperative chemotherapy remains undefined. The authors examine the results of large Phase II and randomized studies as well as the role of neoadjuvant chemotherapy in the context of bladder preservation strategies. Finally, issues of improving therapeutic efficacy and directing clinical research are discussed.


Germinal center reaction in the joints of mice with collagen-induced arthritis: an animal model of lymphocyte activation and differentiation in arthritis joints.
To establish an animal model and provide a basis for investigating the role of germinal center (GC) reaction in autoimmune arthritis. DBA/1 mice were immunized with bovine type II collagen (CII) to elicit collagen-induced arthritis (CIA). Sections of arthritic joints were examined by in situ immunohistochemical studies, and purified cells from affected joints were subjected to flow cytometry analysis. De novo GC reaction was induced in the arthritic joints of male DBA/1 mice by immunization with bovine CII. In comparison with GCs formed in lymphoid tissues, such as spleen and lymph nodes, we found that these GCs formed in the joint tissues of mice with CIA were morphologically typical, as determined by immunohistologic and flow cytometric assays. The local immune responses in murine CIA induced ectopic GC formation, as observed in the synovial tissues of patients with rheumatoid arthritis. This system will allow for the first time the direct study of the role of the GC reaction in autoimmune arthritis in an animal model.


Treatment with an SSRI antidepressant restores hippocampo-hypothalamic corticosteroid feedback and reverses insulin resistance in low-birth-weight rats.
Low birth weight (LBW) is associated with type 2 diabetes and depression, which may be related to prenatal stress and insulin resistance as a result of chronic hypothalamic-pituitary-adrenal (HPA) axis hyperactivity. We examined whether treatment with a selective serotonin reuptake inhibitor [escitalopram (ESC)] could downregulate HPA axis activity and restore insulin sensitivity in LBW rats. After 4-5 wk of treatment, ESC-exposed LBW (SSRI-LBW) and saline-treated control and LBW rats (Cx and LBW) underwent an oral glucose tolerance test or a hyperinsulinemic euglycemic clamp to assess whole body insulin sensitivity. Hepatic phosphoenolpyruvate carboxykinase (PEPCK) mRNA expression and red skeletal muscle PKB Ser(473) phosphorylation were used to assess tissue-specific insulin sensitivity. mRNA expression of the hypothalamic mineralocorticoid receptor was fivefold upregulated in LBW (P < 0.05 vs. Cx), accompanied by increased corticosterone release during restraint stress and total 24-h urinary excretion (P < 0.05 vs. Cx), whole body insulin resistance (P < 0.001 vs. Cx), and impaired insulin suppression of hepatic PEPCK mRNA expression (P < 0.05 vs. Cx). Additionally, there was a tendency for reduced red muscle PKB Ser(473) phosphorylation. The ESC treatment normalized corticosterone secretion (P < 0.05 vs. LBW), whole body insulin sensitivity (P < 0.01) as well as postprandial suppression of hepatic mRNA PEPCK expression (P < 0.05), and red muscle PKB Ser(473) phosphorylation (P < 0.01 vs. LBW). We conclude that these data suggest that the insulin resistance and chronic HPA axis hyperactivity in LBW rats can be reversed by treatment with an ESC, which downregulates HPA axis activity, lowers glucocorticoid exposure, and restores insulin sensitivity in LBW rats.


Comparison of postural ergonomics between laparoscopic and robotic sacrocolpopexy: a pilot study.
To compare resident, fellow, and attending urologic and gynecologic surgeons' musculoskeletal and mental strain during laparoscopic and robotic sacrocolpopexy. Prospective cohort study (Canadian Task Force classification II-2). Academic medical center. Patients who underwent robotic or laparoscopic sacrocolpopexy from October 2009 to January 2011. The Body Part Discomfort (BPD) survey was completed before cases, and the National Aeronautics and Space Administration Task Load Index and BPD survey were completed after cases. Higher scores on BPD and the National Aeronautics and Space Administration Task Load Index indicate greater musculoskeletal discomfort and mental strain. BPD scores were averaged over the following body regions: head/neck, back, hand/wrist, arms, and knees/ankles/feet. Changes in body region-specific discomfort scores were the primary outcomes. Multivariable analysis was performed using mixed-effects linear regression with surgeon as a random effect. Sixteen surgeons participated (53% fellows, 34% residents, and 13% attendings). Thirty-three robotic and 53 laparoscopic cases were analyzed, with a median surgical time of 231 minutes (interquartile range, 204-293 minutes) versus 227 minutes (interquartile range, 203-272 minutes; p = .31), a median estimated blood loss of 100 mL (interquartile range, 50-175 mL) versus 150 mL (interquartile range, 50-200 mL; p = .22), and a mean patient body mass index of 27 ± 4 versus 26 ± 4 kg/m(2) (p = .26), respectively. Robotic surgeries were associated with lower neck/shoulder (-0.19 [interquartile range, -0.32 to -0.01], T = -2.49) and back discomfort scores (-0.35 [interquartile range, -0.58 to 0], T = -2.38) than laparoscopic surgeries. Knee/ankle/foot and arm discomfort increased with case length (0.18 [interquartile range, 0.02-0.3], T = 2.81) and (0.07 [interquartile range, 0.01-0.14], p = .03), respectively. Surgeons performing minimally invasive sacrocolpopexy experienced less neck, shoulder, and back discomfort when surgery was performed robotically.


Fetal urinary tract abnormalities detected on maternal ultrasound.
Twenty-one cases of fetal urinary tract abnormalities were detected on maternal ultrasound in a 5 year period. Six fetuses died and 13 of the 15 surviving cases have had corrective urological surgery, usually during the neonatal period. Maternal ultrasound provides advanced warning of major urinary tract abnormalities in the newborn. Therefore, appropriate investigations and medical management can be instituted early and potentially destructive urological lesions can be corrected as soon as possible. As yet, antenatal intervention does not seem to be warranted as the most common cause of bilateral upper tract dilatation in the surviving cases in this series was primary reflux, and in the cases that died the urinary abnormalities were not salvagable.


Gut microbiota reinforce host antioxidant capacity via the generation of reactive sulfur species.
Gut microbiota act beyond the gastrointestinal tract to regulate the physiology of the host. However, their contribution to the antioxidant capacity of the host remains largely understudied. In this study, we observe that gut bacteria increase the steady-state plasma levels of high-antioxidant molecules, reactive sulfur species (RSS), such as hydrogen sulfide and cysteine persulfide (CysSSH), in the host. Moreover, gut bacteria utilize cystine as a substrate to enzymatically produce CysSSH. Administration of cystine to mice increases their plasma levels of RSS and suppresses the concanavalin-A-induced oxidative stress and liver damage in a gut-microbiota-dependent manner. We find that gut bacteria belonging to the Lachnospiraceae and Ruminococcaceae families have a high capacity to produce RSS, requiring pyridoxal 5'-phosphate for their enzymatic reactions. Collectively, our data demonstrate that gut microbiota enhance the antioxidant capacity of the host through the generation of RSS.


[Development of new in vitro chemosensitivity test using collagen gel droplet embedded culture and its clinical usefulness].
We developed a new in vitro assay for chemosensitivity test using collagen gel droplet embedded culture and image analysis. In this in vitro assay, we successfully minimized the cancer cell number required for culture to approximately 3-10 x 10(3) cells for each 30 microliters collagen gel droplet, obtained the sufficient growth of cancer cells using serum-free medium while suppressing the growth of fibroblastic cells, and measured the volume of cancer cells by eliminating the contaminating fibroblastic cells by an image processing technique. Anticancer effects of the in vitro assay showed a very good correlation with those of in vivo nude mouse assay using human cancer cell lines. The success rates of the in vitro assay for 141 surgical specimens of primary lung cancers and for 65 of primary breast cancers were 89 and 80%, respectively. The accumulated in vitro assay response rates of MMC, CDDP, VDS and VP-16 for primary lung cancers and of MMC, 5-FU and ADR for primary breast cancers were similar to the respective clinical response rates. These results suggest that this in vitro chemosensitivity test may be practically useful for clinical applications.


Garlic (Allium sativum) feeding impairs Sertoli cell junctional proteins in male Wistar rat testis: microscopy study.
Sertoli cell junctions, such as adhesion junction (AJ), gap junction (GJ) and tight junction (TJ), are important for maintaining spermatogenesis. In previous studies, we showed the inhibitory effect of crude garlic (Allium sativum, As) on spermatogenesis and steroidogenesis. The aim of this work was to complete our investigation on the impact of this plant, especially on Sertoli cell junctional proteins (SCJPs). During 1 month, 24 male rats were divided into groups: group control (0% of As) and treated groups fed 5%, 10% and 15% of As. Light and electron microscopy observations were performed to localise junctional proteins: connexin-43, Zona Occluding-1 and N-cadherin (immunohistochemistry) and to describe junctions. We showed that the specific cells involved in the localisation of the SCJP were similar in both control and treated groups, but with different immunoreactivity intensity between them. The electron microscopy observation focused on TJs between Sertoli cells, constituting the blood-testis barrier, showed ultrastructural changes such as fragmentation of TJs between adjacent Sertoli cell membranes and dilatation of rough endoplasmic reticulum saccules giving an aspect of scale to these junctions. We concluded that crude garlic consumption during 1 month induces perturbations on Sertoli cell junctions. These alterations can explain apoptosis in testicular germ cells previously showed.


Tuning quantum corrections and magnetoresistance in ZnO nanowires by ion implantation.
Using ion implantation, the electrical as well as the magnetotransport properties of individual ZnO nanowires (NWs) can be tuned. The virgin NWs are configured as field-effect transistors which are in the enhancement mode. Al-implanted NWs reveal a three-dimensional metallic-like behavior, for which the magnetoresistance is well described by a semiempirical model that takes into account the presence of doping induced local magnetic moments and of two conduction bands. On the other hand, one-dimensional electron transport is observed in Co-implanted NWs. At low magnetic fields, the anisotropic magnetoresistance can be described in the framework of weak electron localization in the presence of strong spin-orbit scattering. From the weak localization, a large phase coherence length is inferred that reaches up to 800 nm at 2.5 K. The temperature-dependent dephasing is shown to result from a one-dimensional Nyquist noise-related mechanism. At the lowest temperatures, the phase coherence length becomes limited by magnetic scattering.


Spouse-rated vs self-rated health as predictors of mortality.
The Health and Retirement Study is a national sample of Americans older than 50 years and their spouses. The present study evaluated cross-sectional and longitudinal data from January 2000 through December 2006. The objective of the study was to evaluate the roles of spouse-rated vs self-rated health as predictors of all-cause mortality among adults older than 50 years. A total of 673 dyads of married couples were randomly selected to participate in a Health and Retirement Study module examining spouse-rated health. For each couple, one member was asked to rate his or her overall health status, and his or her spouse was asked to report the partner's overall health status. Mortality data were available through 2006. Our findings demonstrate that spouse-rated health (area under the curve, 0.75) is as strong a predictor of mortality as self-rated health (area under the curve, 0.73) (chi(2)(1) = 0.36, P = .54). Combining spouse-rated and self-rated health predicts mortality better than using self-rated health alone (area under the curve, 0.77) (chi(2)(1) = 6.72, P = .009). Spouse ratings of health are at least as strongly predictive of mortality as self-rated health. This suggests that, when self-rated health is elicited as a prognostic indicator, spouse ratings can be used when self-ratings are unavailable. Both measures together may be more informative than either measure alone.


[Recent trends in studies of the etiology of hypertension: Endothelium-derived factors].
Endothelial cells play diverse biological roles, such as maintaining vascular tone and structure. Recent progress in vascular biology has revealed that the endothelium releases a large number of vasoactive substances. These substances are divided into two classes: endothelium-derived relaxing factors(EDRFs), and endothelium-derived contracting factors(EDCFs). It has been shown that EDRFs such as nitric oxide(NO) protects vasculature from atherogenic insults, whereas EDCFs, such as endothelin, have opposite effects and participate in the progression of cardiovascular diseases. The exposure of coronary risk factors decreases the bioactivity of EDRFs and increases the release of EDCFs. Endothelial dysfunction caused by imbalance between EDRFs and EDCFs precedes and promotes atherosclerosis by several mechanisms, such as adhesion of monocytes and platelets, increase in vascular permeability, proliferation and migration of smooth muscle cells. In this article, physiology and pathophysiology of these endothelium-derived substances will be reviewed.


Event, incidence and case fatality rates of cerebrovascular disease in Auckland, New Zealand.
Death rates for cerebrovascular disease (stroke) in New Zealand are declining. To investigate the reasons for this decline and to measure the impact of stroke on a defined population, a register of new episodes of stroke was kept in the Auckland region for the year ending March 1982. All cases were followed for one year, with in-depth interviews at onset, one month and six months and a telephone follow-up at one year to establish dead or alive status. A total of 703 episodes were registered for 680 patients, 331 men and 349 women. The crude event rate for all those over 15 years was 228 and 220 per 100,000 for men and women, respectively. Age-adjusted event rates for all strokes were 28% higher for men than women and the age-adjusted event rates for Maoris were 44% higher than for non-Maoris. The case fatality rates were 23.1% at one week, 33.5% at one month, 43.5% at six months and 48.5% at one year. In comparison with other studies, case fatality rates are similar but the incidence rates appear to be lower.


Impact of HPV infection in adolescent populations.
Human papillomavirus (HPV) is a significant source of morbidity and mortality worldwide. The primary risk factors for acquiring HPV are generally associated with sexual activity. Evidence suggests that condoms provide some protection against infection and disease progression, but any genital contact is sufficient for HPV transmission. HPV is so common and transmissible that having just one sexual partner often results in infection. Indeed, cumulative prevalence rates are as high as 82% among adolescent women in select populations. As such, nearly all sexually active adolescents are at high risk for acquiring HPV. Persistent infection with high-risk HPV types (e.g., HPV 16 or 18) is considered necessary for the development of cervical cancer, whereas infection with low-risk HPV types (e.g., HPV 6 or 11) is associated with the development of genital warts and other low-grade genital abnormalities. Most infections are asymptomatic and are efficiently cleared by the immune system. Similarly, both low- and high-grade lesions caused by HPV can regress in adolescent and young adult women. Treatment guidelines allow for observation of adolescent women who develop low-grade lesions rather than immediate colposcopy. Nonetheless, a small percentage of adolescents will develop precancerous lesions that may progress to invasive cervical cancer. Adolescents should be given appropriate education about HPV and the dangers associated with infection; they should also be encouraged to obtain appropriate gynecological care after initiating sexual activity. This article discusses HPV infection and the causal role that HPV plays in the development of low- and high-grade genital lesions, cervical cancer, and genital warts.


CHCHD2 and CHCHD10: Future therapeutic targets in cognitive disorder and motor neuron disorder.
CHCHD2 and CHCHD10 are homolog mitochondrial proteins that play key roles in the neurological, cardiovascular, and reproductive systems. They are also involved in the mitochondrial metabolic process. Although previous research has concentrated on their functions within mitochondria, their functions within apoptosis, synaptic plasticity, cell migration as well as lipid metabolism remain to be concluded. The review highlights the different roles played by CHCHD2 and/or CHCHD10 binding to various target proteins (such as OPA-1, OMA-1, PINK, and TDP43) and reveals their non-negligible effects in cognitive impairments and motor neuron diseases. This review focuses on the functions of CHCHD2 and/or CHCHD10. This review reveals protective effects and mechanisms of CHCHD2 and CHCHD10 in neurodegenerative diseases characterized by cognitive and motor deficits, such as frontotemporal dementia (FTD), Lewy body dementia (LBD), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS). However, there are numerous specific mechanisms that have yet to be elucidated, and additional research into these mechanisms is required.


Neonatal circumcision: anatomic contraindications.
Complications from neonatal circumcision may result from inexperience or poor technique, but may also result from poor patient selection based on penile anatomy. The anatomic contraindications to neonatal circumcision are therefore presented in an effort to prevent complications.


The Rapid Cognitive Screen (RCS): A Point-of-Care Screening for Dementia and Mild Cognitive Impairment.
There is a need for a rapid screening test for mild cognitive impairment (MCI) and dementia to be used by primary care physicians. The Rapid Cognitive Screen (RCS) is a brief screening tool (< 3 min) for cognitive dysfunction. RCS includes 3-items from the Veterans Affairs Saint Louis University Mental Status (SLUMS) exam: recall, clock drawing, and insight. Study objectives were to: 1) examine the RCS sensitivity and specificity for MCI and dementia, 2) evaluate the RCS predictive validity for nursing home placement and mortality, and 3) compare the RCS to the clock drawing test (CDT) plus recall. Patients were recruited from the St. Louis, MO Geriatric Research Education and Clinical Center (GRECC), Veterans Affairs Medical Center (VAMC) hospitals (study 1) or the Saint Louis University Geriatric Medicine and Psychiatry outpatient clinics (study 2). Study 1 participants (N=702; ages 65-92) completed cognitive evaluations and 76% (n=533/706) were followed up to 7.5 years for nursing home placement and mortality. Receiver operator characteristic (ROC) curves were computed to determine sensitivity and specificity for MCI (n=180) and dementia (n=82). Logistic regressions were computed for nursing home placement (n=31) and mortality (n=176). Study 2 participants (N=168; ages 60-90) completed the RCS and SLUMS exam. ROC curves were computed to determine sensitivity and specificity for MCI (n=61) and dementia (n=74). RCS predicted dementia and MCI in study 1 with optimal cutoff scores of ≤ 5 for dementia (sensitivity=0.89, specificity=0.94) and ≤ 7 for MCI (sensitivity=0.87, specificity=0.70). The CDT plus recall predicted dementia and MCI in study 1 with optimal cutoff scores of ≤ 2 for dementia (sensitivity=0.87, specificity=0.85) and ≤ 3 for MCI (sensitivity=0.62, specificity=0.62). Higher RCS scores were protective against nursing home placement and mortality. The RCS predicted dementia and MCI in study 2. The 3-item RCS exhibits good sensitivity and specificity for the detection of MCI and dementia, and higher cognitive function on the RCS is protective against nursing home placement and mortality. The RCS may be a useful screening instrument for the detection of cognitive dysfunction in the primary care setting.


Molecular pathology of prostate cancer: the key to identifying new biomarkers of disease.
Microarray technology has recently accelerated the study of the molecular events involved in prostate cancer, offering the prospect of more precise prognosis and new therapeutic strategies. This review summarises current knowledge of the molecular pathology of prostate cancer. The expression and function of numerous genes have been shown to be altered in prostate cancer. Many of these genes are involved in cell cycle regulation, steroid hormone metabolism or regulation of gene expression. The mechanisms by which androgen independence arises are discussed, including cross-activation, gene amplification and point mutations of the androgen receptor. Analysis of changes in the levels of expression of large numbers of genes during prostate cancer progression have provided a better understanding of the basis of the disease, yielding new molecular markers, such as hepsin, with potential use in diagnosis and prognosis.


The alternatively folded state of the antibody C(H)3 domain.
The C(H)3 domain of antibodies is characterized by two antiparallel beta-sheets forming a disulfide-linked sandwich-like structure. At acidic pH values and low ionic strength, C(H)3 becomes completely unfolded. The addition of salt transforms the acid-unfolded protein into an alternatively folded state exhibiting a characteristic secondary structure. The transition from native to alternatively folded C(H)3 is a fast reaction. Interestingly, this reaction involves the formation of a defined oligomer consisting of 12-14 subunits. Association is completely reversible and the native dimer is quantitatively reformed at neutral pH. This alternatively folded protein is remarkably stable against thermal and chemical denaturation and the unfolding transitions are highly cooperative. With a t(m) of 80 degrees C, the stability of the alternatively folded state is comparable to that of the native state of C(H)3. The defined oligomeric structure of C(H)3 at pH 2 seems to be a prerequisite for the cooperative unfolding transitions.


Clinical findings and diagnostic tests in Creutzfeldt-Jakob disease and variant Creutzfeldt-Jakob disease.
Sporadic Creutzfeldt-Jakob disease (sCJD) is a rare transmissible disease caused by accumulation of pathological prion protein in the CNS. sCJD typically affects patients in their sixties. The median disease duration in sCJD (6 months) is shorter than in variant Creutzfeldt-Jakob disease (vCJD) (14 months). The clinical diagnosis in sCJD is supported by the detection of periodic sharp and slow wave complexes (PSWC) in the electroencephalogram, 14-3-3 proteins in the cerebrospinal fluid (CSF) and hyperintense basal ganglia on magnetic resonance imaging (MRI). In contrast to sCJD, hyperintensities in the posterior pulvinar (the "pulvinar sign") are seen in vCJD. Different sCJD subtypes characterised by distinct neuropathological lesion profiles, clinical features and codon 129 genotype of the prion protein gene (PRNP) are described, together with the type with a proteinase K-resistant core of the prion protein. The sensitivity of diagnostic tests varies considerably in different sCJD subtypes. Alzheimer 's disease and Lewy body dementia are the most frequent differential diagnoses in elderly patients, while chronic inflammatory CNS disorders have to be considered in younger patients.


Building an online community to support nurse education.
This article explores the topic of developing an online community for student nurses to use in learning. It examines the different definitions and types of e-learning and outlines the online community's role in healthcare education, together with some of its pitfalls. A comparison is then made to the process of bidding on eBay, to determine possible similarities.


Computationally enhanced quantitative phase microscopy reveals autonomous oscillations in mammalian cell growth.
The fine balance of growth and division is a fundamental property of the physiology of cells, and one of the least understood. Its study has been thwarted by difficulties in the accurate measurement of cell size and the even greater challenges of measuring growth of a single cell over time. We address these limitations by demonstrating a computationally enhanced methodology for quantitative phase microscopy for adherent cells, using improved image processing algorithms and automated cell-tracking software. Accuracy has been improved more than twofold and this improvement is sufficient to establish the dynamics of cell growth and adherence to simple growth laws. It is also sufficient to reveal unknown features of cell growth, previously unmeasurable. With these methodological and analytical improvements, in several cell lines we document a remarkable oscillation in growth rate, occurring throughout the cell cycle, coupled to cell division or birth yet independent of cell cycle progression. We expect that further exploration with this advanced tool will provide a better understanding of growth rate regulation in mammalian cells.


[Antigenic localities in the tissues of the young adult worm of Paragonimus westermani using immunogold labeling method].
In order to observe the antigenic localization in the tissues of the young adult Paragonimus westermani, immunogold labeling method was applied using serum immunoglobulins(IgG) of the dog which infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size; 12 nm). It was observed by electron microscopy at each tissues of the worm. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were predominantly labeling on the epithelial lamela and lumen of caecum. The above finding showed that antigenic materials in young adult worm tissue were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells.


Heart rhythm disturbances in the inhabitants of mountainous regions.
The authors studied 513 males, permanently living in the high-mountain regions of Tian Shan and the Pamirs (2800 - 4000 m above sea level). A control group consisted of 404 males permanently living at low altitudes (780-900 m above sea level) in the Kemin District, Kirghiz SSR. The probands' ages were 30-59 years. In all of them the resting electrocardiograms were recorded; 110 exercise tests were made in the high mountains, and 35 tests, at the low altitudes. The prevalence of heart rhythm disturbances was statistically significantly higher in the inhabitants of the high-mountain regions (12.1%) than in the low-altitude inhabitants (2.9%; p less than 0.0001). The most frequent disturbance was the 1st-degree a-v block (6 per cent). In the high mountains cardiac arrhythmias are usually associated with right ventricular hypertrophy, caused by high-altitude hypoxia. During exercise heart arrhythmias appeared conspicuously less frequently in the high mountain than in the low altitude inhabitants.


HPV 6-positive giant keratoacanthoma in an immunocompetent patient.
Keratoacanthoma (KA) is a clinically distinct, rapidly growing lesion that generally presents as a solitary crateriform nodule in sun-exposed areas in elderly, fair-skinned individuals. A KA larger than 20-30 mm is referred to as giant keratoacanthoma, a relatively rare lesion especially in young patients. Such lesions grow rapidly with possible destruction of underlying tissues. In addition to ultraviolet exposure, KAs have also been associated with chemical carcinogens, chemical peels, genetic factors, chronic skin conditions that produce scarring, trauma and thermal burns. Immunosuppressed patients, especially after transplantation, also develop KAs. A viral etiology has been suggested but not confirmed. We encountered a case of giant keratoacanthoma (greater than 50 mm in diameter) with induration of underlying structures on the upper lip of a 39-year-old male sailor. The patient reported sudden appearance and rapid enlargement of the lesion in only three weeks. Biopsy of the cutaneous lesion and the characteristic clinical history suggested the diagnosis of keratoacanthoma. Total excision with primary closure of the defect by a nasolabial advancement flap was performed. Histological examination of the tumor mass confirmed the diagnosis of KA with infiltrative growth and perineural invasion. Immunosuppression was excluded by blood analyses, as were HIV, syphilis and hepatitis infections. Only low-risk genital HPV type 6 was detected in the lesion, suggesting a possible cocarcinogenic effect of HPV and UV light in a chronically sun-exposed patient.


MicroRNA-20a-5p regulates the epithelial-mesenchymal transition of human hepatocellular carcinoma by targeting RUNX3.
MicroRNA-20a (miR-20a) is dysregulated in many types of malignancies, including human hepatocellular carcinoma (HCC), but its expression level and functional significance in HCC are still disputed. We aimed to study the role of miR-20a-5p in HCC and its downstream molecular mechanisms. We used real-time polymerase chain reaction to detect the expression of miR-20a-5p and runt-related transcription factor 3 ( RUNX3 ) in HCC and paraneoplastic tissue, transfected Huh7 and highly metastatic human hepatocellular carcinoma (MHCC97H) cells. A live cell workstation was used to observe the proliferation and migration of transfected cells. The invasiveness of transfected cells was verified by Transwell assay. Cell apoptosis was detected by flow cytometry. The expression levels of proteins after transfection were measured using simple western immunoblot measurements. Gene expression profiles between HCC and normal samples were obtained from The Cancer Genome Atlas. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment results were processed by the database for annotation, visualization and integrated discovery. Potential target genes of miR-20a-5p were predicted to further investigate how miR-20a-5p regulates epithelial-mesenchymal transition (EMT) in HCC. MiR-20a-5p was significantly highly expressed in HCC tissues, and overexpression of miR-20a-5p significantly promoted HCC cell proliferation, migration, and invasion and inhibited apoptosis in vitro. The protein expression of E-cadherin was decreased and that of vimentin was increased after overexpression of miR-20a-5p in HCC cells. We discovered the intersection of genes from miRDB, miR TarBase, and TargetScan, obtained 397 target genes and finally focused on RUNX3. RUNX3 was not only reduced in HCC specimens but also drastically reduced in HCC cells overexpressing miR-20a-5p. RUNX3 expression decreased with elevated miR-20a-5p, which activated downstream EMT signaling and promoted cell proliferation, migration, and invasion. Since RUNX3 is involved in EMT in HCC, as proven by previous research, our findings provide further evidence for a novel regulatory pathway comprising the miR-20a/RUNX3/EMT axis that upregulates EMT signaling and enhances the migration of HCC cells.


Change in the amount of epsilon-hexosyllysine, UV absorbance, and fluorescence of collagen with age in different animal species.
Skin and aorta collagen specimens of Wistar rats, white mice, beagle dogs, cats, horses, and human necropsies of different ages were examined with respect to the content of glycated products. The data presented show that (a) glycation and accumulation of the chromophore(s) are comparable in collagen samples from different species of comparable age; (b) glycation and pigmented accumulation increase markedly during the first 5-10 years of age; (c) the extent of glycation is different in different tissues (in particular, glycation of aortal collagen is about twice that of skin collagen); and (d) collagen pigmentation as followed by fluorescence is comparable in aortal and skin collagen (except below 10 years); pigmentation measured by absorbance at 350 nm is, on the contrary, lower in aortal than in skin collagen. Based on the assumption of constant blood glucose level during the life span, it appears feasible to conclude that the degree of nonenzymatic collagen glycation reflects the time period for which the protein was exposed to the action of sugars. This period, because of increased cross-linking, is likely to be extended in older animals. Other factors, such as differences in collagen turnover between different tissues and the intensity of the removal process of the glycated products, should be taken into consideration as well.


Destructive colon injuries requiring resection: Is colostomy ever indicated?
The management of destructive colon injuries requiring resection has shifted from mandatory diverting stoma to liberal use of primary anastomosis. Various risk criteria have been suggested for the selection of patients for primary anastomosis or ostomy. At our center, we have been practicing a policy of liberal primary anastomosis irrespective of risk factors. The purpose of this study was to evaluate the colon-related outcomes in patients managed with this policy. This retrospective study included all colon injuries requiring resection. Data collected included patient demographics, injury characteristics, blood transfusions, operative findings, operations performed, complications, and mortality. A total of 287 colon injuries were identified, 101 of whom required resection, forming the study population. The majority (63.4%) were penetrating injuries. Furthermore, 16.8% were hypotensive on admission, 40.6% had moderate or severe fecal spillage, 35.6% received blood transfusion of >4 U, and 41.6% had Injury Severity Score of >15. At index operation, 88% were managed with primary anastomosis and 12% with colon discontinuity, and one patient had stoma. Damage-control laparotomy (DCL) with temporary abdominal closure was performed in 39.6% of patients. Of these patients with DCL, 67.5% underwent primary anastomosis, 30.0% were left with colon discontinuity, and 2.5% had stoma. Overall, after the definitive management of the colon, including those patients who were initially left in colon discontinuity, only six patients (5.9%) had a stoma. The incidence of anastomotic leaks in patients with primary anastomosis at the index operation was 8.0%, and there was no colon-related mortality. The incidence of colon anastomotic leaks in the 27 patients with DCL and primary anastomosis was 11.1%, and there was no colon-related mortality. Multivariate analysis evaluating possible risk factors identified discontinuity of the colon as independent risk factor for mortality. Liberal primary anastomosis should be considered in almost all patients with destructive colon injuries requiring resection, irrespective of risk factors. Therapeutic/Care Management; Level IV.


[Tarsus-plasty with chondroplasty].
For a few years new Chondroplast has been available as a ready-to-use product in a variety of different shapes and sizes. The material is obtained from bovine cartilage and is made antigen-free and stable against decomposition by treatment with glutaraldehyde and irradiation. To date we have used Chondroplast for lid stabilization in 9 eyes with very slack entropion or ectropion. For tarsusplasty secondary to tumor resection we chose Chondroplast for seven eyes. A thin 0.5-mm-thick lamella was cut out of the original material and positioned in a preformed pocket between the orbicularis muscle and the skin and conjunctiva and then fixed with Vicryl sutures. Corrective of eyelid malposition was combined with pentagonal excision for ectropion, or resection of the orbicularis muscle for entropion. In plastic lid replacement the material was sutured to the existing wound edges of the lid margin and covered posteriorly with conjunctiva and anteriorly with advancement or transposed flaps. In all patients the implant took well, and there were no complications during wound healing. The cosmetic and functional results were satisfactory. The largest post-operation follow-up time was over 1 year. In one case--1 year after resection of a lower eyelid basalioma--we had the opportunity to examine histologically the tissue in the area of a Chondroplast implant. We performed this biopsy excision because of a suspected recurrence. Macroscopically the implanted cartilage lamella was surrounded by a barely vascularized capsule. Histologically we found a non-vital cartilage implant surrounded by a mainly fibrotic connective tissue capsule. In only a few places was there evidence of a slight reaction to the foreign body but without substantial inflammation activity. There was also no histological indication of resorption of the implant.


Expression of certain antiproliferative and growth-related genes in isolated mouse pancreatic islets: analysis by polymerase chain reaction.
With the aim at understanding the mechanism responsible for the low proliferative capacity of the adult pancreatic B-cell, expression of the "antiproliferative" gene retinoblastoma and that coding for p53 as well as certain genes coding for cyclin B1, cyclin D1 and p34cdc2 and p33cdk2 kinases, all of which are important for the cell cycle and mitosis, was assessed by the polymerase chain reaction in a semiquantitative assay. Islet expression of the p53 and retinoblastoma genes was higher than that of the liver, but lower or the same as that of the spleen, making expression of these genes an unlikely explanation for the low replicatory capacity of the B-cell. Similarly, islet expression of the genes coding for p33cdk2 kinase and cyclin D1 was not very different from that of the spleen. The levels of cyclin B1 mRNA and the mRNA coding for the p34cdc2 kinase were however both low in islets, compared with those of the spleen and the insulin-producing RINm5F cell line. Stimulation of B-cell proliferation did not change the expression of any of the genes studied. It is concluded that the low expression of cyclin B1 and p34cdc2 kinase and failure to induce these by stimulation of B-cell replication may play a role in maintaining adult B-cells in a state of low proliferative capacity.


Construction and Performance of Visible SISAM and SISAM X F. P. Monochromators.
In a previous paper [Appl. Opt. 12, 279 (1973)] new methods for scanning and modulating a SISAM monochromator were described. These made it useful in the visible domain. Here a developed version of the previous crude prototype is described and its performance evaluated. Further a SISAM and a Fabry-Perot have been combined and used in the visible. The performance of this monochromator is also described.


NMR quantum computing: applying theoretical methods to designing enhanced systems.
Density functional theory results for chemical shifts and spin-spin coupling constants are presented for compounds currently used in NMR quantum computing experiments. Specific design criteria were examined and numerical guidelines were assessed. Using a field strength of 7.0 T, protons require a coupling constant of 4 Hz with a chemical shift separation of 0.3 ppm, whereas carbon needs a coupling constant of 25 Hz for a chemical shift difference of 10 ppm, based on the minimal coupling approximation. Using these guidelines, it was determined that 2,3-dibromothiophene is limited to only two qubits; the three qubit system bromotrifluoroethene could be expanded to five qubits and the three qubit system 2,3-dibromopropanoic acid could also be used as a six qubit system. An examination of substituent effects showed that judiciously choosing specific groups could increase the number of available qubits by removing rotational degeneracies in addition to introducing specific conformational preferences that could increase (or decrease) the magnitude of the couplings. The introduction of one site of unsaturation can lead to a marked improvement in spectroscopic properties, even increasing the number of active nuclei.