Projections of thalamic gustatory and lingual areas in the monkey, Macaca fascicularis.
The efferent projections of the parvicellular division of the ventroposteromedial nucleus of the thalamus (VMPpc; thalamic taste area) were traced to cortex in Macaca fascicularis by using tritiated amino acid autoradiography. Labeled fascicles could be traced from VPMpc to two discrete regions of cortex. The primary efferent projection was located on ipsilateral insular-opercular cortex adjacent to the superior limiting sulcus and extended as far rostrally as the posterior lateral orbitofrontal cortex. An additional projection was located within primary somatosensory (SI) cortex subjacent to the anterior subcentral sulcus. Following autoradiographic injections in VPM, the trigeminal somatosensory relay, a dense terminal plexus was labeled on SI cortex of both pre- and postcentral gyri, but not within insular-opercular cortex. The autoradiographic data were verified by injecting each cortical projection area with horseradish peroxidase (HRP) and observing the pattern of retrogradely labeled somata within the thalamus. Injections in the precentral gyrus near the anterior subcentral sulcus retrogradely labeled neurons within VPMpc, whereas injections further caudally near the floor of the central sulcus labeled neurons within VPM. Injections of HRP within opercular, insular, or posterior lateral orbitofrontal cortex retrogradely labeled neurons within VPMpc.


Prediction of postoperative trunk imbalance after posterior spinal fusion with pedicle screw fixation for adolescent idiopathic scoliosis.
The objective of this retrospective study was to explore, which radiographic parameters, immediately after posterior spinal fusion with pedicle screw fixation for adolescent idiopathic scoliosis (AIS), best correlate with subjacent disc wedging at a minimum of 2-year follow-up. Sixty-four consecutive AIS patients who underwent posterior pedicle screw-only instrumentation were studied. Preoperative and postoperative radiographs were obtained to measure various parameters regarding global coronal, shoulder, sagittal, and regional balance. Specific correlation of these parameters to selected 2-year postoperative disc wedging and lowest-instrumented vertebra (LIV) tilt and translation were analyzed. The average lateral disc opening changed from 4.59±4.75 preoperatively to 1.46±2.82 at 2 weeks and 2.81±6.43 at 2 years postoperatively. Two-year postoperative lateral disc opening significantly correlated with the 2-week postoperative lateral disc opening, C7 plumbline relative to the posterior-superior corner of the first sacral vertebra distance, and LIV-center sacral vertical line (CSVL) distance (r=0.7433, P<0.0001). The two-year postoperative LIV tilt significantly correlated with the 2-week postoperative LIV tilt, T12-LIV lordosis, LIV-CSVL distance, and C7-CSVL distance (r=0.8879, P<0.0001). Two-year postoperative LIV-CSVL significantly correlated with 2-week postoperative LIV-CSVL distance and lateral disc opening (r=0.6104, P<0.0001). Two-year postoperative disc wedging, LIV tilt, and LIV translation occurred most often when disc wedging and LIV deviation or obliquity existed immediately postoperatively. This study identified a potential indicator for AIS repair. Preoperative surgical planning and intraoperative correction are important for avoiding subjacent regional imbalance after scoliosis fusion.


Overexpression of leucine-rich α2-glycoprotein-1 is a prognostic marker and enhances tumor migration in gastric cancer.
Gastric cancer is one of the most common malignant tumors. Although improvement in chemotherapy has been achieved, the clinical prognosis of advanced gastric cancer remains poor. Therefore, it is increasingly important to predict the prognosis and determine whether patients should or should not receive neoadjuvant or adjuvant chemotherapy. Leucine-rich α2-glycoprotein-1 (LRG1) is overexpressed during inflammation and is associated with various malignancies. In this study, we assessed LRG1 expression in cancer specimens and in the sera of patients with cancer to clarify the usefulness of LRG1 as a biomarker in gastric cancer. This study enrolled 239 (for immunohistochemical staining; IHC) and 184 (for ELISA) patients with gastric cancer. Results of IHC showed that LRG1 expression was significantly associated with histological type, lymphatic and venous invasion, tumor and node factors, and disease stage. Overall survival was significantly worse in the high LRG1 expression group than in the low LRG1 group (P = 0.0003). Cox multivariate analysis of overall survival revealed that LRG1 expression was an independent prognostic factor (P = 0.0258). Serum LRG1 was significantly higher in gastric cancer patients than in healthy volunteers, and increased as the pathological stage progressed. Furthermore, a significant correlation was revealed between serum LRG1 level and LRG1 expression with IHC (P < 0.0001). Inhibition of LRG1 significantly decreased cell proliferation in vitro (migratory and invasive capacity of gastric cancer cells). These results suggest that LRG1 expression in tumors and serum may be a useful prognostic marker in gastric cancer patients.


Function of hormone-sensitive lipase in diacylglycerol-protein kinase C pathway.
To explore the functional effects of hormone-sensitive lipase (HSL) in diacylglycerol (DAG) metabolism, Chinese hamster ovary cells were stably transfected with rat HSL cDNA (wt-HSL), inactive mutant S423A-HSL cDNA (S423A) and pcDNA3 vector alone (Ct). [(14)C]Glucose-incorporation into triglyceride (TG) was 75% lower in the presence or absence of insulin in cells expressing wt-HSL compared to Ct or S423A. [(14)C]Glucose-incorporation into DAG was 33% lower without insulin and 51% lower with insulin in cells expressing wt-HSL compared to Ct or S423A. Insulin stimulated glucose-incorporation into DAG 2.2-fold in S423A and Ct cells, whereas only a 50% increase was observed in cells expressing wt-HSL. Phospholipase C-mediated release of DAG from membrane phospholipids was reduced 70% in cells expressing wt-HSL compared to Ct or S423A. Western blot analysis showed that membrane-bound protein kinase C (PKC)-alpha and -epsilon were decreased 40-50% in cells expressing wt-HSL grown in high glucose with insulin. These data show that HSL potentially hydrolyzes cellular DAG generated either by de novo synthesis from glucose or release from membrane phospholipids by phospholipase C, resulting in a reduction in the translocation of DAG-sensitive PKCs.


Growth-differentiation factor 15 and risk of major bleeding in atrial fibrillation: Insights from the Randomized Evaluation of Long-Term Anticoagulation Therapy (RE-LY) trial.
To evaluate and validate the prognostic value of growth-differentiation factor 15 (GDF-15) beyond clinical characteristics and other biomarkers concerning bleeding and stroke outcomes in patients with atrial fibrillation in the RE-LY trial. GDF-15 was measured in samples collected at randomization in 8,474 patients with a median follow-up time of 1.9 years. Patients were stratified based on predefined GDF-15 cutoffs: group 1, <1,200 ng/L (the 90th percentile in healthy individuals); group 2, 1,200-1,800; and group 3, >1,800 ng/L (high-risk individuals). Efficacy and safety outcomes were compared across groups of GDF-15 in Cox models adjusted for baseline characteristics, cardiac (N-terminal pro-b-type natriuretic peptide, high-sensitive troponin T), inflammatory (interleukin 6, C-reactive protein) and coagulation (D-dimer) biomarkers, and randomized treatment. GDF-15 concentrations were <1,200 ng/L in 2,647 (31.2%), between 1,200 and 1,800 ng/L in 2,704 (31.9%), and >1,800 ng/L in 3,123 (36.9%) participants, respectively. Annual rates of stroke, major bleeding, and mortality increased with higher GDF-15 levels. The prognostic value of GDF-15 was independent of clinical characteristics for these outcomes. In models also adjusted for biomarkers, GDF-15 remained significantly associated with major bleeding (hazard ratio [95% CI] group 3 vs group 1 1.76 [1.28-2.42], P < .0005) and all-cause mortality (hazard ratio 1.72 [1.30-2.29], P < .0005). GDF-15 improved the c index of both the HAS-BLED (0.62-0.69) and ORBIT (0.68-0.71) bleeding risk scores. In patients with atrial fibrillation, GDF-15 is an independent risk indicator for major bleeding and all-cause mortality, but not for stroke. Therefore, GDF-15 seems useful as a specific marker of bleeding in patients with AF on oral anticoagulant treatment.


Subcellular distribution and developmental change of 2',3'-cyclic nucleotide 3'-phosphohydrolase in the central nervous system of the myelin-deficient shiverer mutant mice.
Shiverer is an autosomal recessive mutant characterized by dysmyelination in the central nervous system. The myelin in the shiverer is devoid of the major dense line. The pattern of increase of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) in the various parts of central nervous system during postnatal development was identical with that of controls. Subcellular fractionation of the brain from shiverer showed that the recovery of CNPase activity of the P2A was only 14% of the control, but those of P2B and P3 were several times higher than the controls. The specificity activity of CNPase of the purified myelin and P2A from shiverer did not differ significantly from that of the controls. The combined P2 and P3 (P2 + P3) fraction was subjected to linear sucrose density gradient (0.32--1.3 M) centrifugation. By measuring CNPase activities of the fractions obtained after centrifugation, shiverer showed two peaks of CNPase activities in the sucrose gradient, although that of the control showed a single peak. The main peak obtained from shiverer was found in the region of higher sucrose concentration than that of the control, while a small peak was found in the region of lower sucrose concentration.


Neutralization, by a monospecific Bothrops lanceolatus antivenom, of toxic activities induced by homologous and heterologous Bothírops snake venoms.
A monospecific Bothrops lanceolatus antivenom, currently used in Martinique, was tested for its efficacy in the neutralization of several toxic and enzymatic activities of the venoms of B. lanceolatus, B. atrox and B. asper. When tested by the i.p. route in mice, B. lanceolatus venom had an LD50 of 12.8 microg/g. In addition, it induced local tissue damage (hemorrhage, edema and myotoxicity) and showed indirect hemolytic activity, but was devoid of coagulant effect on human plasma in vitro and of defibrinating activity in mice. Antivenom was fully effective in the neutralization of lethal, hemorrhagic, edema-forming, myotoxic and indirect hemolytic effects of B. lanceolatus venom in assays involving preincubation of venom and antivenom. When tested against the venoms of B. asper and B. atrox, the antivenom completely neutralized the lethal, hemorrhagic, myotoxic and indirect hemolytic effects, and was partially effective in neutralizing edema-forming activity. In contrast, the antivenom was ineffective in the neutralization of in vitro coagulant and in vivo defibrinating effects induced by these two venoms.


Curcumin inhibits the mammalian target of rapamycin-mediated signaling pathways in cancer cells.
Curcumin (diferuloylmethane), a polyphenol natural product of the plant Curcuma longa, is undergoing early clinical trials as a novel anticancer agent. However, the anticancer mechanism of curcumin remains to be elucidated. Here we show that curcumin inhibited growth of rhabdomyosarcoma cells (Rh1 and Rh30) (IC50 = 2-5 microM) and arrested cells in G1 phase of the cell cycle. Curcumin also induced apoptosis and inhibited the basal or type I insulin-like growth factor-induced motility of the cells. At physiological concentrations (2.5 microM), curcumin rapidly inhibited phosphorylation of the mammalian target of rapamycin (mTOR) and its downstream effector molecules, p70 S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4E-BP1), in a panel of cell lines (Rh1, Rh30, DU145, MCF-7 and Hela). Curcumin also inhibited phosphorylation of Akt in the cells, but only at high concentrations (>40 microM). The data suggest that curcumin may execute its anticancer activity primarily by blocking mTOR-mediated signaling pathways in the tumor cells.


Comparison of Minocycline Susceptibility Testing Methods for Carbapenem-Resistant Acinetobacter baumannii.
Treatment options for infections due to carbapenem-resistant Acinetobacter baumannii are extremely limited. Minocycline is a semisynthetic tetracycline derivative with activity against this pathogen. This study compared susceptibility testing methods that are used in clinical microbiology laboratories (Etest, disk diffusion, and Sensititre broth microdilution methods) for testing of minocycline, tigecycline, and doxycycline against 107 carbapenem-resistant A. baumannii clinical isolates. Susceptibility rates determined with the standard broth microdilution method using cation-adjusted Mueller-Hinton (MH) broth were 77.6% for minocycline and 29% for doxycycline, and 92.5% of isolates had tigecycline MICs of ≤2 μg/ml. Using MH agar from BD and Oxoid, susceptibility rates determined with the Etest method were 67.3% and 52.3% for minocycline, 21.5% and 18.7% for doxycycline, and 71% and 29.9% for tigecycline, respectively. With the disk diffusion method using MH agar from BD and Oxoid, susceptibility rates were 82.2% and 72.9% for minocycline and 34.6% and 34.6% for doxycycline, respectively, and rates of MICs of ≤2 μg/ml were 46.7% and 23.4% for tigecycline. In comparison with the standard broth microdilution results, very major rates were low (∼2.8%) for all three drugs across the methods, but major error rates were higher (∼5.6%), especially with the Etest method. For minocycline, minor error rates ranged from 14% to 37.4%. For tigecycline, minor error rates ranged from 6.5% to 69.2%. The majority of minor errors were due to susceptible results being reported as intermediate. For minocycline susceptibility testing of carbapenem-resistant A. baumannii strains, very major errors are rare, but major and minor errors overcalling strains as intermediate or resistant occur frequently with susceptibility testing methods that are feasible in clinical laboratories.


A randomized trial of an online, coach-assisted self-management PTSD intervention tailored for women veterans.
Scalable, efficiently delivered treatments are needed to address the needs of women Veterans with PTSD. This randomized clinical trial compared an online, coach-assisted cognitive behavioral intervention tailored for women Veterans with PTSD to phone monitoring only. Women Veterans who met diagnostic criteria for PTSD were randomized to an 8-week web-based intervention, called DElivery of Self TRaining and Education for Stressful Situations (DESTRESS)-Women Veterans version (WV), or to phone monitoring only (N = 102). DESTRESS-WV consisted of online sessions and 15-min weekly phone calls from a study coach. Phone monitoring included 15-min weekly phone calls from a study coach to offer general support. PTSD symptom severity (PTSD Symptom-Checklist-Version 5 [PCL-5]) was evaluated pre and posttreatment, and at 3 and 6 months posttreatment. More participants completed phone monitoring than DESTRESS-WV (96% vs. 76%, p = 0.01), although treatment satisfaction was significantly greater in the DESTRESS-WV condition. We failed to confirm the superiority of DESTRESS-WV in intent-to-treat slope changes in PTSD symptom severity. Both treatments were associated with significant reductions in PTSD symptom severity over time. However, post hoc analyses of treatment completers and of those with baseline PCL ≥ 33 revealed that the DESTRESS-WV group had greater improvement in PTSD symptom severity relative to phone monitoring with significant differences at the 3-month follow-up assessment. Both DESTRESS-WV and phone monitoring resulted in significant improvements in women Veterans' PTSD symptoms. DESTRESS-WV may be an appropriate care model for women Veterans who can engage in the demands of the treatment and have higher baseline symptoms. Future research should explore characteristics of and the methods of reliably identifying women Veterans who are most likely to benefit. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Ulinastatin inhibits renal tubular epithelial apoptosis and interstitial fibrosis in rats with unilateral ureteral obstruction.
The effect of ulinastatin (UTI) on renal tubular epithelial apoptosis and interstitial fibrosis in rats with unilateral ureteral obstruction (UUO) was investigated. A total of 18 male Wistar rats were randomly divided into the following 3 groups: The Sham group (n=6), the UUO group (n=6), and the UTI group (n=6). In the UUO and UTI groups, the left ureter was ligated to establish a UUO model. Starting from day 1 after surgery, an intervention treatment was performed using normal saline (1 ml/kg/d) and UTI (40,000 unit/kg/d). On day 7 after surgery, 6 rats from each group were sacrificed. In the Sham group, the left ureter was only freed, not ligated; after 7 days of abdominal closure, all of the rats were sacrificed. Blood samples were collected prior to sacrificing the animals to measure the blood urea nitrogen (BUN) and serum creatinine (Scr). The incidence of renal interstitial lesions on the obstruction side was observed by hematoxylin and eosin, and Masson staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, and immunohistochemical detection of apoptosis regulator Bax (Bax), apoptosis regulator Bcl‑2 (Bcl‑2) and caspase‑3 were performed to observe the presence of renal tubular epithelial cell apoptosis. The UTI did not have a significant influence on the mouse BUN and Scr levels in any of the groups (P>0.05). Compared with that in the Sham group, renal tissue injury in the UUO group was significantly aggravated with renal tubular dilation, epithelial cell atrophy, renal interstitial inflammatory cell infiltration and fibrous tissue hyperplasia (P<0.01). Furthermore, the renal tubular epithelial TUNEL+ cell number and Bax and caspase‑3 levels were increased, and the expression of Bcl‑2 was decreased (P<0.01). Following the UTI treatment, the renal interstitial injury at the obstruction side was significantly attenuated (P<0.05), the renal tubular epithelial TUNEL+ cell number, and Bax and caspase‑3 levels significantly decreased, and the expression of Bcl‑2 was restored (P<0.05). UTI inhibited renal tubular epithelial apoptosis and interstitial fibrosis in UUO rats.


Hydration of protein-protein interfaces.
We present an analysis of the water molecules immobilized at the protein-protein interfaces of 115 homodimeric proteins and 46 protein-protein complexes, and compare them with 173 large crystal packing interfaces representing nonspecific interactions. With an average of 15 waters per 1000 A2 of interface area, the crystal packing interfaces are more hydrated than the specific interfaces of homodimers and complexes, which have 10-11 waters per 1000 A2, reflecting the more hydrophilic composition of crystal packing interfaces. Very different patterns of hydration are observed: Water molecules may form a ring around interfaces that remain "dry," or they may permeate "wet" interfaces. A majority of the specific interfaces are dry and most of the crystal packing interfaces are wet, but counterexamples exist in both categories. Water molecules at interfaces form hydrogen bonds with protein groups, with a preference for the main-chain carbonyl and the charged side-chains of Glu, Asp, and Arg. These interactions are essentially the same in specific and nonspecific interfaces, and very similar to those observed elsewhere on the protein surface. Water-mediated polar interactions are as abundant at the interfaces as direct protein-protein hydrogen bonds, and they may contribute to the stability of the assembly.


Treatment of hirsutism by finasteride and flutamide in women with polycystic ovary syndrome.
Forty-four hirsute patients with polycystic ovary syndrome were randomly treated with finasteride (5 mg daily) or flutamide (250 mg twice daily) for 6 consecutive months. Hirsutism was evaluated before and after therapy with the Ferriman-Gallwey score and with measurement of hair diameter (micron). The hairs were taken from four different body areas: the face, abdomen, thighs and forearm. The measurement was carried out with a micrometer applied to the optical microscope. Mean plasma concentrations of luteinizing hormone, follicle-stimulating hormone, 17 alpha-hydroxyprogesterone, androstenedione, testosterone, free testosterone, dehydroepiandrosterone sulfate, insulin, and sex hormone binding globulin were determined before and after therapy. Hematochemical examinations and side-effects were controlled after the treatment. After 6-months' therapy, both antiandrogens significantly reduced the Ferriman-Gallwey score and hair diameter in all the body areas. Finasteride reduced the Ferriman-Gallwey score by 25% and hair diameter by 16-25%; flutamide reduced the score by 20% and hair diameter by 15.3-22%. Abdominal hairs were more sensitive to both drugs. Flutamide induced a significant drop in total testosterone and dehydroepiandrosterone sulfate. No important side-effect or change in the hematochemical parameters was observed. Our data demonstrate that finasteride and flutamide are effective in the treatment of hirsutism in patients with polycystic ovary syndrome.


B7-H1 overexpression regulates epithelial-mesenchymal transition and accelerates carcinogenesis in skin.
B7-H1 (CD274) is a T-cell coinhibitory molecule that is also often induced on human carcinoma cells, where its expression has been implicated in immune escape. Under inflammatory conditions, B7-H1 is also inducible in normal epithelial cells but little is known about its involvement in conversion of normal cells to tumor cells. Here, we show that skin-specific expression of B7-H1 accelerates inflammatory carcinogenesis in a methylcholantrene (MCA)-induced model of squamous cell carcinoma (SCC). Inflammatory responses induced by MCA or phorbol ester TPA were clearly inhibited in B7-H1 transgenic mice (B7-H1tg mice). Antibody-mediated blockade of either B7-H1 or the related molecule PD-1 revealed that their ability to limit inflammation relied on ligand interactions made by B7-H1 or PD-1. Skin keratinocytes derived from B7-H1tg mice exhibited constitutive reduction of E-cadherin, and SCC induced in B7-H1tg mice also showed loss of E-cadherin along with elevated expression of the transcription factors Slug and Twist that drive epithelial-mesenchymal transition (EMT). Our results indicate that upregulation of B7-H1 in skin epithelial cells promotes EMT and accelerates carcinogenesis, revealing insights into the significance of B7-H1 overexpression on solid tumor cells and hinting at a close relationship between EMT and immune escape signaling pathways in cancer.


Effect of norepinephrine on ketogenesis, fatty acid oxidation, and esterification in isolated rat hepatocytes.
Recent studies in man demonstrated a marked ketogenic effect of increased plasma norepinephrine concentrations as observed in diabetic ketoacidosis. Since this effect may have been due either to increased substrate supply for ketogenesis (lipolysis) or to direct hepatic activation of ketogenesis, the latter mechanism was examined in isolated rat hepatocytes. Incubation of hepatocytes with norepinephrine (10(-7) to 10(-4) M) resulted in a dose-dependent increase in conversion of the long-chain fatty acid [1-14C]palmitate into ketone bodies and CO2. Norepinephrine decreased [1-14C]palmitate conversion into triglycerides without affecting fatty acid uptake. Norepinephrine enhanced ketogenesis from [1-14C]palmitate in a physiologic range of fatty acid concentrations (0.5-2.5 mM), but failed to affect fatty acid esterification to phospholipids or mono- and diglycerides. In contrast to long-chain fatty acids, oxidation of the medium-chain fatty acid [1-14C]octanoate to ketone bodies was not enhanced by norepinephrine, whereas CO2 production increased. The effect of norepinephrine on [1-14C]fatty acid oxidation was blocked by the alpha 1 receptor blocker prazosin. The results demonstrate that norepinephrine diverts long-chain fatty acids into the pathways of oxidation and ketogenesis away from esterification, suggesting enhanced carnitine-dependent mitochondrial fatty acid uptake. The studies using octanoate indicated that norepinephrine also enhanced fatty acid oxidation by increasing the flux of acetyl-CoA through the Krebs cycle. The data suggest that stress-associated sympathetic activation and norepinephrine discharge, as observed in diabetic ketoacidosis, result in direct activation of ketogenesis in the liver.


Analgesia induced by dietary restriction is mediated by the kappa-opioid system.
Progress in the control and treatment of pain may be facilitated by a better understanding of mechanisms underlying nociceptive processing. Here we show that mice subjected to an intermittent fasting diet (IFD) display markedly reduced responses in models of thermal and visceral pain compared with mice fed ad libitum (AL). Pharmacological analyses suggest that a change in the endogenous kappa-opioid system underlies IFD-induced analgesia. The levels of prodynorphin mRNA and kappa-opioid receptors in the spinal cord are higher in IFD than in AL mice. Furthermore, in spinal cord nuclear protein extracts, the activity of the transcriptional repressor DREAM (downstream regulatory element antagonist modulator), the main regulator of prodynorphin expression, is lower in IFD than in AL mice. Finally, c-Fos expression in dorsal spinal cord after noxious stimulation is significantly lower in IFD than in AL animals, indicating that dynorphin could block nociceptive information at the spinal cord. These results suggest that dietary restriction together with administration of kappa-opioid agonists could be useful as a new therapeutic approach for pain relief.


The Detection of Dopamine Gene Receptors (DRD1-DRD5) Expression on Human Peripheral Blood Lymphocytes by Real Time PCR.
There is interrelationship between the immune and nervous systems that is accomplished by the molecular mediators. Dopamine is one of the most important neurotransmitters. Five different dopamine receptor genes (DRD1, DRD2, DRD3, DRD4, and DRD5) have been recognized and cloned. The expression of the dopamine receptors is well characterized in the brain but little work has been done to examine their expression in other organ tissues. In certain diseases of the immune and nervous systems, alterations in dopamine receptors gene expression in different cells have been reported. This suggests that dopamine and its receptors have important role in pathophysiology of above-mentioned diseases.In the present study, using Real Time Polymerase Chain Reaction (PCR) technique, we investigated dopamine receptors genes expression in PBMC of normal individuals. The PBMC was separated from normal whole blood by Ficoll-hypaque; the total cellular RNA was then extracted and the cDNA was synthesized. This process followed by real time-PCR using primer pairs specific for five dopamine receptors mRNAs and beta-actin as internal control. The results showed the presence of all types of dopamine receptors in lymphocytes of normal individuals. The specificities of the obtained PCR products for the respective dopamine receptors fragments were confirmed by sequenced analysis capillary system. In conclusion, the present study has shown that human lymphocytes express five dopamine receptors DR1-DR5. However, the conclusive evidence on the possible function of these receptors in lymphocytes remains unknown. Because lymphocytes express all of the five neuronal dopamine receptors, it is quite reasonable to consider them as a model of dopaminergic neuron.


Evaluation of the Physical, Chemical, Technological, and Sensorial Properties of Extrudates and Cookies from Composite Sorghum and Cowpea Flours.
In recent years, there has been a growing demand for gluten-free and functional products, driven by consumer preferences for healthier and more diverse food choices. Therefore, there is a need to explore new ingredients that can be used as alternatives to traditional gluten-containing grains. Thus, this work evaluated the physical, chemical, technological, and sensorial properties of extrudates and cookies from composite tannin sorghum (rich in resistant starch) and white cowpea flours. Extrudates and cookies were produced from a composite flour made of sorghum and cowpea, at a sorghum:cowpea flour ratio of 70:30, 50:50, and 30:70. Then, raw flours, cookies, and extrudates were characterized (dietary fiber, resistant starch, proteins, antioxidant capacity, pasting properties, etc.). Results obtained for particle size distribution and bulk density indicated that the particles increased and the color changed with the addition of cowpea flour. The raw tannin sorghum flour had a higher resistant starch concentration (36.3%) and antioxidant capacity (211.2 µmolTE/g), whereas cowpea flour had higher levels of proteins (18.7%) and dietary fiber (20.1%). This difference in the raw flour composition contributed to the nutritional value of the extrudates and cookies, especially the cookies which undergo dry heat and had higher retention of resistant starch and antioxidants. Moreover, sorghum flour presented a higher tendency to retrograde (high setback), which was decreased by the addition of cowpea flour. Overall acceptance and intention to purchase were higher for extrudates with 100% sorghum flour (6.52 and 68.3%, respectively) and cookies with 70% cowpea flour (7.03 and 76.7%, respectively). Therefore, nutritious and functional gluten-free extrudates and cookies, of good acceptability, can be produced from composite tannin sorghum and white cowpea flours.


Vector semirational rogue waves and modulation instability for the coupled higher-order nonlinear Schrödinger equations in the birefringent optical fibers.
We report the existence and properties of vector breather and semirational rogue-wave solutions for the coupled higher-order nonlinear Schrödinger equations, which describe the propagation of ultrashort optical pulses in birefringent optical fibers. Analytic vector breather and semirational rogue-wave solutions are obtained with Darboux dressing transformation. We observe that the superposition of the dark and bright contributions in each of the two wave components can give rise to complicated breather and semirational rogue-wave dynamics. We show that the bright-dark type vector solitons (or breather-like vector solitons) with nonconstant speed interplay with Akhmediev breathers, Kuznetsov-Ma solitons, and rogue waves. By adjusting parameters, we note that the rogue wave and bright-dark soliton merge, generating the boomeron-type bright-dark solitons. We prove that the rogue wave can be excited in the baseband modulation instability regime. These results may provide evidence of the collision between the mixed ultrashort soliton and rogue wave.


Combined quantitation of HMGA2 mRNA, microRNAs, and mitochondrial-DNA content enables the identification and typing of thyroid tumors in fine-needle aspiration smears.
Analysis of molecular markers in addition to cytological analysis of fine-needle aspiration (FNA) samples is a promising way to improve the preoperative diagnosis of thyroid nodules. Nonetheless, in clinical practice, applications of existing diagnostic solutions based on the detection of somatic mutations or analysis of gene expression are limited by their high cost and difficulties with clinical interpretation. The aim of our work was to develop an algorithm for the differential diagnosis of thyroid nodules on the basis of a small set of molecular markers analyzed by real-time PCR. A total of 494 preoperative FNA samples of thyroid goiters and tumors from 232 patients with known histological reports were analyzed: goiter, 105 samples (50 patients); follicular adenoma, 101 (48); follicular carcinoma, 43 (28); Hürthle cell carcinoma, 25 (11); papillary carcinoma, 121 (56); follicular variant of papillary carcinoma, 80 (32); and medullary carcinoma, 19 (12). Total nucleic acids extracted from dried FNA smears were analyzed for five somatic point mutations and two translocations typical of thyroid tumors as well as for relative concentrations of HMGA2 mRNA and 13 microRNAs and the ratio of mitochondrial to nuclear DNA by real-time PCR. A decision tree-based algorithm was built to discriminate benign and malignant tumors and to type the thyroid cancer. Leave-p-out cross-validation with five partitions was performed to estimate prediction quality. A comparison of two independent samples by quantitative traits was carried out via the Mann-Whitney U test. A minimum set of markers was selected (levels of HMGA2 mRNA and miR-375, - 221, and -146b in combination with the mitochondrial-to-nuclear DNA ratio) and yielded highly accurate discrimination (sensitivity = 0.97; positive predictive value = 0.98) between goiters with benign tumors and malignant tumors and accurate typing of papillary, medullary, and Hürthle cell carcinomas. The results support an alternative classification of follicular tumors, which differs from the histological one. The study shows the feasibility of the preoperative differential diagnosis of thyroid nodules using a panel of several molecular markers by a simple PCR-based method. Combining markers of different types increases the accuracy of classification.


Quantifying soil carbon stocks and humification through spectroscopic methods: A scoping assessment in EMBU-Kenya.
A soil carbon assessment was performed comparing agricultural cropping systems with natural vegetation along a sampling transect spanning different agro-ecologies on the eastern foot slopes of Mount Kenya in Embu county, 125 km from Nairobi, Kenya. The aim was to determine differences in soil carbon stocks and carbon recalcitrance and relate these to soil textural class, altitude, climatic parameters and land use. Soils from main agricultural systems as tea, coffee and maize-based intercropping, as well as from natural vegetation cover were sampled in triplicates, in five layers from 0 to 30 cm in depth and processed for total carbon analysis. The whole soil samples were also analysed using Laser-Induced Fluorescence Spectroscopy (LIFS) to assess carbon humification. Prototype portable equipment intended for future in situ analysis was used in the lab to ascertain the structure of the most recalcitrant and stable carbon present in different agro-ecosystems. In addition, Near Infrared Spectroscopy (NIRS) was tested for the quantitative analysis of soil carbon, showing that it is a reproducible and low-cost method that provided satisfactory results under the processing conditions of the samples. Results showed wide variation in the level and quality of carbon stored in the soils, depending on soil texture, land use, elevation, climate, agricultural practices and land use history. Considering the heterogeneous nature of sampled soils and the performance of NIRS and LIFS, these results can be used as a basis for the development of fully portable systems able to provide rapid, clean and potentially cost-effective relevant information for soil management.


Self-transcendence and well-being in homeless adults.
This study examines the relationships of spiritually and physically related variables to well-being among homeless adults. A convenience sample of 61 sheltered homeless persons completed the Spiritual Perspective Scale, the Self-Transcendence Scale, the Index of Well-Being, and items measuring fatigue and health status. The data were subjected to correlational and multiple regression analysis. Positive, significant correlations were found among spiritual perspective, self-transcendence, health status, and well-being. Fatigue was inversely correlated with health status and well-being. Self-transcendence and health status together explained 59% of the variance in well-being. The findings support Reed's theory of self-transcendence, in which there is the basic assumption that human beings have the potential to integrate difficult life situations. This study contributes to the growing body of evidence that conceptualizes homeless persons as having spiritual, emotional, and physical capacities that can be used by health care professionals to promote well-being in this vulnerable population.


[Uncertainty, relationship with agencies and asymmetrical information (II)].
In the second part of this article, the author relates the "knightian" uncertainty to consumer, i.e. patient, preferences and to types of choice, considering that choice in a health organization usually is made based on uncertain, or high risk, expectations whose determination is often difficult. The author reviews the concepts of 1) "ethical risk"--or potencial overuse of a form of treatment when the consumer does not bear its cost directly--, 2) "asymmetrical information", and 3) "adverse choice"; all of which are related to relationships with an agency and therefore, have the corresponding costs derived from these agencies themselves. Finally, the author reviews the "agency theory" and, in relationships to the agency itself, evaluates the "agency costs", costs which are not always evaluated in a just manner. The author make use of the same flight comparison, British Airways flight number 1, as he did in the first part of this article to bring these concepts into health field and to provide a clearer description of these concepts.


Assessment and consequences of the constant-volume attribute of the four-chambered heart.
The constant-volume hypothesis regarding the four-chambered heart states that total pericardial volume remains invariant throughout the cardiac cycle. Previous canine studies have indicated that the pericardial volume remains constant within 5%; however, this hypothesis has not been validated in humans using state-of-the-art technology. The constant-volume hypothesis has several predictable functional consequences, including a relationship between atrial ejection fraction and chamber equilibrium volumes. Using cardiac magnetic resonance (MR) imaging (MRI), we measured the extent to which the constant-volume attribute of the heart is valid, and we tested the accuracy of the predicted relationship between atrial ejection fraction and chamber equilibrium volumes. Eleven normal volunteers and one volunteer with congenital absence of the pericardium were imaged using a 1.5-T MR scanner. A short-axis cine-loop stack covering the entire heart was acquired. The cardiac cycle was divided into 20 intervals. For each slice and interval, pericardial volumes were measured. The slices were stacked and summed, and total pericardial volume as a function of time was determined for each subject. In the normal subjects, chamber volumes at ventricular end diastole, end systole, and diastasis were measured. Pericardial volume remained invariant within 5 +/- 1% in normal subjects; maximum variation occurred near end systole. In the subject with congenital absence of the pericardium, total heart volume, defined by the epicardial surface, varied by 12%. The predictions of the relationship between atrial ejection fraction and chamber equilibrium volumes were well fit by MRI data. In normal subjects, the four-chambered heart is a constant-volume pump within 5 +/- 1%, and constant-volume-based modeling accurately predicts previously unreported physiological relationships.


On deducing causality in metabolic networks.
Metabolic networks present a complex interconnected structure, whose understanding is in general a non-trivial task. Several formal approaches have been developed to support the investigation of such networks. One of the relevant problems in this context is the comprehension of causality dependencies amongst the molecules involved in the metabolic process. We apply techniques from formal methods and computational logic to develop an abstract qualitative model of metabolic networks in order to determine possible causal dependencies. Keeping in mind both expressiveness and ease of use, we aimed at providing: i) a minimal notation to represent causality in biochemical interactions, and ii) an automated tool allowing human experts to easily vary conditions of in silico experiments. We exploit a reading of chemical reactions in terms of logical implications: starting from a description of a metabolic network in terms of reaction rules and initial conditions, chains of reactions, causally depending one from the another, can be automatically deduced. Both the components of the initial state and the clauses ruling reactions can be easily varied and a new trial of the experiment started, according to a what-if investigation strategy. Our approach aims at exploiting computational logic as a formal modeling framework, amongst the several available, that is naturally close to human reasoning. It directly leads to executable implementations and may support, in perspective, various reasoning schemata. Indeed, our abstractions are supported by a computational counterpart, based on a Prolog implementation, which allows for a representation language closely correspondent to the adopted chemical abstract notation. The proposed approach has been validated by results regarding gene knock-out and essentiality for a model of the metabolic network of Escherichia coli K12, which show a relevant coherence with available wet-lab experimental data. Starting from the presented work, our goal is to provide an effective analysis toolkit, supported by an efficient full-fledged computational counterpart, with the aim of fruitfully driving in vitro experiments by effectively pruning non promising directions.


Visible-Light-Driven CO2 Reduction with Homobimetallic Complexes. Cooperativity between Metals and Activation of Different Pathways.
Visible-light-driven reduction of CO2 to both CO and formate (HCOO-) was achieved in acetonitrile solutions using a homobimetallic Cu bisquaterpyridine complex. In the presence of a weak acid (water) as coreactant, the reaction rate was enhanced, and a total of ca. 766 TON (turnover number) was reached for the CO2 reduction, with 60% selectivity for formate and 28% selectivity for CO, using Ru(phen)32+ as a sensitizer and amines as sacrificial electron donors. Mechanistic studies revealed that with the help of cooperativity between two Cu centers, a bridging hydride is generated in the presence of a proton source (water) and further reacts with CO2 to give HCOO-. A second product, CO, was also produced in a parallel competitive pathway upon direct coordination of CO2 to the reduced complex. Mechanistic studies further allowed comparison of the observed reactivity to the monometallic Cu quaterpyridine complex, which only produced CO, and to the related homobimetallic Co bisquaterpyridine complex, that has been previously shown to generate formate following a mechanism not involving the formation of an intermediate hydride species.


Metatarsalgia.
Metatarsalgia (ie, metatarsal pain) is one of the most common reports in patients with foot problems. This pain is confined to the area across the plantar forefoot, including the second through fourth metatarsal heads. However, it is frequently accompanied by deformity of the first and fifth rays as well as of the toes. There is great variability in possible causative factors, but all of them seem to be related to gait mechanics, foot anatomy, and foot and ankle deformity. An individualized treatment protocol is required. Nonsurgical management is usually sufficient to achieve satisfactory results. Surgical correction must be precise, and all pain-producing deformities must be corrected. Most patients present with abnormalities of the distal metatarsals. Metatarsal osteotomy, long a staple of treatment, always fails in the long term. Improved equipment and internal fixation methods may lead to better long-term outcomes.


Antioxidant responses of two barley varieties to saline stress.
In this study, we investigated antioxidant responses of activities of Superoxide Dismutase (SOD), Catalase (CAT), Ascorbate Peroxidase (APX) and Guaiacol Peroxidase (GPX) to saline stress in two barley varieties named Hordeum vulgare L. var. Afzal and var. EMB82-12 treated with 50, 100, 200, 300 and 400 mM NaCl for 3 days. The MDA content of Afzal plants grown under different salt regimes remained nearly constant but it largely increased in EMB82-12 plants under the same conditions. There was a linear and significant correlation in CAT, APX, SOD, GPX activities in Afzal plants in response to increased salt concentration. The strong and positive correlation between antioxidant enzymes and salt concentrations, may account for the MDA level of Afzal plants remaining constant in response to different salt regimes. In general, the activities of antioxidant enzymes were increased in the root and shoot under saline stress. But the increase was more significant and consistent in the root. Among the antioxidant enzymes, CAT activity was increased the most drastically.


Activity recognition using dynamic multiple sensor fusion in body sensor networks.
Multiple sensor fusion is a main research direction for activity recognition. However, there are two challenges in those systems: the energy consumption due to the wireless transmission and the classifier design because of the dynamic feature vector. This paper proposes a multi-sensor fusion framework, which consists of the sensor selection module and the hierarchical classifier. The sensor selection module adopts the convex optimization to select the sensor subset in real time. The hierarchical classifier combines the Decision Tree classifier with the Naïve Bayes classifier. The dataset collected from 8 subjects, who performed 8 scenario activities, was used to evaluate the proposed system. The results show that the proposed system can obviously reduce the energy consumption while guaranteeing the recognition accuracy.


Distribution and determinants of pesticide mixtures in cord serum using principal component analysis.
We characterized the distribution and determinants of fetal exposures to pesticide mixtures using a cross-sectional study of 297 singletons delivered at Johns Hopkins Hospital in Baltimore, MD (2004-2005). Concentrations of nine persistent and twelve nonpersistent pesticides were measured in cord serum. Mixtures were identified using principal components analysis. Associations between mixtures and maternal and infant characteristics were evaluated using multivariate analysis. p,p'-DDE, p,p'-DDT, trans-nonachlor, oxychlordane, bendiocarb, propoxur, and trans- and cis-permethrin were detected in 100, 90, 93, 84, 73, 55, 52, and 41% of serum samples, respectively. There were four independent pesticide components: DDT (p,p'-DDT + p,p'-DDE), chlordane (trans-nonachlor + oxychlordane), permethrin (trans- and cis-permethrins + PBUT), and carbamate (bendiocarb + propoxur). DDT and chlordane were 6.1 (95%CI: 2.4, 15.5) and 2.1 (95%CI: 1.0, 4.2) times higher for infants of women >35, and 1.8 (95%CI: 1.2, 2.9) and 1.5 (95%CI: 1.1, 2.1) times higher in smoking mothers. DDT and carbamate were 15 (95%CI: 7, 30) and 2 (95%CI: 1, 4) times higher for infants of Asian compared with Caucasian mothers. No significant differences were observed for permethrin. Fetal exposures to pesticides are widespread, occur as mixtures, and differ by maternal race, age, and smoking status.


Ambulatory surgery for anorectal diseases: an update.
More than 90% of anorectal surgery can be done in an outpatient setting. The savings for anorectal procedures in 1982 for inpatient room cost alone could have been almost $200 million. The use of local anesthesia for anorectal surgery enhances patient safety. Ambulatory surgery should be considered whenever anorectal procedures are contemplated.


The potency of KUL-7211, a selective ureteral relaxant, in isolated canine ureter: comparison with various spasmolytics.
We compared the potency of a selective ureteral relaxant KUL-7211 (beta(2)/beta(3)-adrenoceptor agonist; (-)-2-[4-(2-{[(1S,2R)-2-hydroxy-2-(4-hydroxyphenyl)-1-methylethyl]amino}ethyl)phenyloxy]acetic acid) with those of various spasmolytics on contractions in isolated canine ureteral preparations. Drug effects were evaluated on the tonic contraction induced by KCl (80 mM) and on spontaneous, 1x10(-5) M phenylephrine-, and 1x10(-6) M PGF(2alpha)-induced rhythmic contractions in isolated canine ureteral preparations using a functional experimental technique. The potencies (pD(2) value) of the following drugs were compared: KUL-7211, tamsulosin (an alpha(1A/1D)-adrenoceptor antagonist), prazosin (an alpha(1)-adrenoceptor antagonist), verapamil (a Ca(2+)-channel blocker), butylscopolamine (a nonselective muscarinic antagonist), and papaverine (a phosphodiesterase inhibitor). The rank order of relaxing potencies against KCl-induced tonic contraction was KUL-7211 (6.60)>tamsulosin(5.90)>verapamil(5.70)>papaverine(4.88)>prazosin (4.54). The rank order of potencies for reductions in spontaneous rhythmic contractions was KUL-7211 (6.80)>verapamil(6.12)>papaverine(5.05). Conversely, high concentrations of the two alpha-adrenoceptor antagonists (tamsulosin and prazosin) and of butylscopolamine enhanced the spontaneous contractions, although at low concentrations (up to 1x10(-6) M) they had no significant effects. For suppression of spasmogen-induced rhythmic contractions, the rank order of potencies was, against phenylephrine-induced contractions: KUL-7211 (6.95)>tamsulosin(6.26)>prazosin(5.68)>verapamil(5.64)>papaverine (5.03), and against PGF(2alpha)-induced contractions: KUL-7211 (7.05)>verapamil(6.70)>papaverine (5.27). Our results suggest that in dogs, the beta(2)/beta(3)-adrenoceptor agonist KUL-7211 is the most efficacious ureteral relaxant among the spasmolytics tested against various contractions. Possibly, KUL-7211 might be useful for promoting stone passage and relieving ureteral colic in urolithiasis patients.


A magnetic vehicle realized tumor cell-targeted radiotherapy using low-dose radiation.
Radiotherapy, a common cancer treatment, often adversely affects the surrounding healthy tissue and/or cells. Some tumor tissue-focused radiation therapies have been developed to lower radiation-induced lesion formation; however, achieving tumor cell-targeted radiotherapy (i.e., precisely focusing the radiation efficacy to tumor cells) remains a challenge. In the present study, we developed a novel tumor cell-targeted radiotherapy, named targeted sensitization-enhanced radiotherapy (TSER), that exploits tumor-specific folic acid-conjugated carboxymethyl lauryl chitosan/superparamagnetic iron oxide (FA-CLC/SPIO) micelles to effectively deliver chlorin e6 (Ce6, a sonosensitizer) to mitochondria of HeLa cells under magnetic guidance. For the in vitro tests, the sensitization of Ce6 induced by ultrasound, that could weaken the radiation resistant ability of tumor cells, occurred only in Ce6-internalizing tumor cells. Therefore, low-dose X-ray irradiation, that was not harmful to normal cells, could exert high tumor cell-specific killing ability. The ratio of viable normal cells to tumor cells was increased considerably, from 7.8 (at 24h) to 97.1 (at 72h), after they had received TSER treatment. Our data suggest that TSER treatment significantly weakens tumor cells, resulting in decreased viability in vitro as well as decreased in vivo subcutaneous tumor growth in nude mice, while the adverse effects were minimal. Taken together, TSER treatment appears to be an effective, clinically feasible tumor cell-targeted radiotherapy that can solve the problems of traditional radiotherapy and photodynamic therapy.


Role of Toll-like receptor 4 and human defensin 5 in primary endocervical epithelial cells.
Endocervical epithelial cells play early roles in the defense of upper female genital tract to pathogens. Toll-like receptors (TLRs) and human defensins (HD) have recently been identified as fundamental components of the innate immune responses to bacterial pathogens. We aimed to use in vitro model of human primary endocervical epithelial cells (HPECs) to investigate their roles in innate immune response of the endocervix. TLR4 expression and distribution in HPECs and endocervix were investigated by immunofluorescence (IF). Cultured HPECs were divided into lipopolysaccharide (LPS) group which were treated by LPS for 0, 24 and 48 hours, and control group without treatment. At each time point, the levels of HD5, IL-6 and TNF-alpha in supernants were determined by ELISA. TLR4 and HD5 expressions of cells were detected by Western blotting simultaneously. HD5 expression pattern was also compared between the HeLa cell line and HPECs. Endocervix tissue surface and HPECs expressed TLR4. After incubated with LPS, HPECs expressed significantly higher levels of TLR4 than control group, especially after 24 hours (P < 0.01), however decreased after 48 hours with a similar level of TLR4 expression compared with control group. LPS could upregulate the secretion of HD5, IL-6 and TNF-alpha in a time-dependent manner (24 hours: P < 0.05; 48 hours: P < 0.01, compared with control group). Intracellular HD5 expression levels decreased over time. HD5 expression patterns in HPECs were different from HeLa cell line. To respond to LPS stimulation, HPECs may function in the mucosal immune defense through TLR4 activation and HD5 secretion. HPEC is considered a significant model for immunological study.


Network changes in patients with phobic postural vertigo.
Functional dizziness comprises a class of dizziness disorders, including phobic postural vertigo (PPV), that cause vestibular symptoms in the absence of a structural organic origin. For this reason, functional brain mechanisms have been implicated in these disorders. Here, functional network organization was investigated in 17 PPV patients and 18 healthy controls (HCs) during functional magnetic resonance imaging with a visual motion stimulus, data initially collected and described by Popp et al. (2018). Graph theoretical measures (degree centrality [DC], clustering coefficient [CC], and eccentricity) of 160 nodes within six functional networks were compared between HC and PPV patients during visual motion and static visual patterns. Graph theoretical measures analyzed during the static condition revealed significantly different DC in the default-mode, sensorimotor, and cerebellar networks. Furthermore, significantly different group differences in network organization changes between static visual and visual motion stimulation were observed. In PPV, DC and CC showed a significantly stronger increase in the sensorimotor network during visual stimulation, whereas cerebellar network showed a significantly stronger decrease in DC. These results suggest that the altered visual motion processing seen in PPV patients may arise from a modified state of sensory and cerebellar network connectivity.


Orbitomaxillofacial Mucormycosis Requiring Complex Multifactorial Management.
Mucormycosis is a rare fungal infection caused by ubiquitous fungi in the order Mucorales. It is the most rapidly progressing fulminant fungal infection that mimics necrotizing soft-tissue infections. Overwhelming fungal sepsis develops quickly and mortality rates approach 70%. Culture negative necrotizing infections and cutaneous necrosis following a vascular pattern should raise suspicion for this rare entity. We describe avoiding mortality in a case of orbitomaxillofacial mucormycosis multifactorially treated with: radical serial debridement, topical amphotericin B irrigation and dressings, parenteral amphotericin B, and hyperbaric oxygen therapy. Tissue biopsy was central to confirming the diagnosis and directing multimodal management that ultimately prevented dissemination to the central nervous system and mortality.


Evolution of dispersal and life history interact to drive accelerating spread of an invasive species.
Populations on the edge of an expanding range are subject to unique evolutionary pressures acting on their life-history and dispersal traits. Empirical evidence and theory suggest that traits there can evolve rapidly enough to interact with ecological dynamics, potentially giving rise to accelerating spread. Nevertheless, which of several evolutionary mechanisms drive this interaction between evolution and spread remains an open question. We propose an integrated theoretical framework for partitioning the contributions of different evolutionary mechanisms to accelerating spread, and we apply this model to invasive cane toads in northern Australia. In doing so, we identify a previously unrecognised evolutionary process that involves an interaction between life-history and dispersal evolution during range shift. In roughly equal parts, life-history evolution, dispersal evolution and their interaction led to a doubling of distance spread by cane toads in our model, highlighting the potential importance of multiple evolutionary processes in the dynamics of range expansion.


Alar Contour Grafts in Rhinoplasty: A Safe and Reproducible Way to Refine Alar Contour Aesthetics.
Alar rim deformities such as retraction, notching, collapse, and asymmetry are common problems in rhinoplasty patients. Although alar rim deformities may be improved through rhinoplasty, this area is prone to late changes because of scarring of the soft triangles and a paucity of native structural support. The purpose of this study was to analyze the effect of alar contour grafts on primary rhinoplasty. Fifty consecutive primary rhinoplasty patients with preoperative and postoperative photographs who received alar contour grafts were evaluated for alar aesthetics; 50 consecutive primary rhinoplasty patients without such grafts served as controls. Differences among alar retraction, notching, collapse, and asymmetry from anterior, lateral, and basal views were evaluated. Follow-up ranged from 1 to 4 years and was graded on a four-point scale. The average difference between the two groups' aggregate preoperative scores was 0.21 (p = 0.24). The average preoperative and postoperative scores in the nongraft group were significant for worsening retraction, notching, and collapse but insignificant for asymmetry. The preoperative and postoperative scores for the graft group were insignificant for retraction but improved significantly for notching, collapse, and asymmetry. Postoperatively, the aggregate average of the scores in the nongroup was 0.32 points worse (p < 0.01), whereas the graft group had a 0.33-point improvement (p < 0.01). Alar contour grafts have a clear and important impact on cosmetic results of primary rhinoplasty. Use of alar contour grafts has been shown to improve aesthetics, whereas there is a worsening of the measured parameters postoperatively without use of these grafts. Therapeutic, III.


Regulation of the mitochondrial matrix volume in vivo and in vitro. The role of calcium.
The ability of alpha-adrenergic agonists and vasopressin to increase the mitochondrial volume in hepatocytes is dependent on the presence of extracellular Ca2+. Addition of Ca2+ to hormone-treated cells incubated in the absence of Ca2+ initiates mitochondrial swelling. In the presence of extracellular Ca2+, A23187 (7.5 microM) induces mitochondrial swelling and stimulates gluconeogenesis from L-lactate. Isolated liver mitochondria incubated in KCl medium in the presence of 2.5 mM-phosphate undergo energy-dependent swelling, which is associated with electrogenic K+ uptake and reaches an equilibrium when the volume has increased to about 1.3-1.5 microliter/mg of protein. This K+-dependent swelling is stimulated by the presence of 0.3-1.0 microM-Ca2+, leading to an increase in matrix volume at equilibrium that is dependent on [Ca2+]. Ca2+-activated K+-dependent swelling requires phosphate and shows a strong preference for K+ over Na+, Li+ or choline. It is not associated with either uncoupling of mitochondria or any non-specific permeability changes and cannot be produced by Ba2+, Mn2+ or Sr2+. Ca2+-activated K+-dependent swelling is not prevented by any known inhibitors of plasma-membrane ion-transport systems, nor by inhibitors of mitochondrial phospholipase A2. Swelling is inhibited by 65% and 35% by 1 mM-ATP and 100 microM-quinine respectively. The effect of Ca2+ is blocked by Ruthenium Red (5 micrograms/ml) at low [Ca2+]. Spermine (0.25 mM) enhanced the swelling seen on addition of Ca2+, correlating with its ability to increase Ca2+ uptake into the mitochondria as measured by using Arsenazo-III. Mitochondria derived from rats treated with glucagon showed less swelling than did control mitochondria. In the presence of Ruthenium Red and higher [Ca2+], the mitochondria from hormone-treated animals showed greater swelling than did control mitochondria. These data imply that an increase in intramitochondrial [Ca2+] can increase the electrogenic flux of K+ into mitochondria by an unknown mechanism and thereby cause swelling. It is proposed that this is the mechanism by which alpha-agonists and vasopressin cause an increase in mitochondrial volume in situ.


Incidental Detection of Hairy Cell Leukaemia with Herpes Simplex Virus (HSV) Related Lip Ulcer Mimicking Carcinoma.
Hairy cell leukemia is a chronic lympho-proliferative disease. It is indolent but progressive in nature. It arises from B-cell lineage. We report an incidentally detected case of Hairy Cell Leukaemia (HCL) in a 55-year-old male patient with Herpes simplex virus (HSV) - related lip ulcer mimicking squamous cell carcinoma. Clinically the patient presented with lip ulceration without pain. He was found to have moderate hepatosplenomegaly and pancytopenia on general examination. Bone marrow aspiration and flow cytometric immunophenotyping revealed HCL. The oral lesion resolved after antiviral therapy. The intriguing possibility of a combined pathogenesis for the two disorders is considered, as HCL is known to be associated with immunosuppression, second malignancies and the production of cytokines promoting epithelial growth.


Cefepime effectiveness in Gram-negative bloodstream infections.
We used a retrospective cohort to evaluate the 28 day all-cause mortality in adult patients with Gram-negative bloodstream infection (BSI) who received cefepime therapy compared with those who received other β-lactam antibiotics with in vitro activity against aerobic Gram-negative bacilli. We identified 398 adult patients who received β-lactam antibiotic monotherapy during hospitalization at Mayo Clinic hospitals in Rochester, MN, USA, for monomicrobial Gram-negative BSI from 1 January 2001 to 31 October 2006. After adjusting for the propensity to receive cefepime, multivariable Cox proportional hazard regression was used to compare the 28 day mortality in patients who were treated with cefepime with that in those who received other β-lactam antibiotics. The median age of patients with Gram-negative BSI was 65 years (interquartile range 51-77) and 230 (58%) were male. Compared with patients who received other β-lactams, patients treated with cefepime were more likely to have cancer (59% versus 44%, P = 0.007) and immunocompromising conditions (55% versus 21%, P < 0.001). Patients who received cefepime were less likely to have community-acquired infection acquisition (18% versus 33%, P = 0.002) and urinary source of BSI (14% versus 23%, P = 0.04) than those treated with other β-lactam antibiotics. After adjusting for the propensity to receive cefepime and other covariates in the multivariable Cox model, cefepime therapy was not associated with an increased 28 day all-cause mortality (hazard ratio 0.99, 95% confidence interval 0.53-1.79, P = 0.97). In adult patients with monomicrobial Gram-negative BSI, cefepime therapy was not associated with increased mortality when compared with other β-lactam antibiotics.


Focus Quality Assessment of High-Throughput Whole Slide Imaging in Digital Pathology.
One of the challenges facing the adoption of digital pathology workflows for clinical use is the need for automated quality control. As the scanners sometimes determine focus inaccurately, the resultant image blur deteriorates the scanned slide to the point of being unusable. Also, the scanned slide images tend to be extremely large when scanned at greater or equal 20X image resolution. Hence, for digital pathology to be clinically useful, it is necessary to use computational tools to quickly and accurately quantify the image focus quality and determine whether an image needs to be re-scanned. We propose a no-reference focus quality assessment metric specifically for digital pathology images that operate by using a sum of even-derivative filter bases to synthesize a human visual system-like kernel, which is modeled as the inverse of the lens' point spread function. This kernel is then applied to a digital pathology image to modify high-frequency image information deteriorated by the scanner's optics and quantify the focus quality at the patch level. We show in several experiments that our method correlates better with ground-truth z -level data than other methods, which is more computationally efficient. We also extend our method to generate a local slide-level focus quality heatmap, which can be used for automated slide quality control, and demonstrate the utility of our method for clinical scan quality control by comparison with subjective slide quality scores.


The morphological identification of pathogenic yeasts using carbohydrate media.
Eight isolates of C. albicans were used to determine the frequency with which germ tube formation occurred: on rice extract -Tween 80 agar, on its components, and on 1% bactopeptone agar after three hr at 37 degrees C; in 0.5% aqueous solution of various carbohydrates; in various concentrations of glucose; on 0.5 and 0.1% glucose agar and on various types of agar alone. Subsequently 250 isolates of yeast of the genera Candida, Torulopsis, Trichosporon, Cryptococcus, and Saccharomyces, which were obtained from a clinical laboratory, were spread on rice extract -Tween 80 agar and on 0.1% glucose agar and covered with coverslips. Direct microscopic examination after incubation for three hours at 37 degrees C demonstrated germ tube formation by all 140 isolates of C. albicans, but by none of the other yeasts. The characteristic features of the pseudomycelia of isolates of Candida and Trichosporon were evident on reexamination after a further 45 to 69 hours at room temperature (22 degrees C). These morphological observations suggested the identity of the isolates of Torulopsis, Cryptococcus, and Saccharomyces but identified virtually all (98.2%) of those of the genera which formed pseudomycelia. Of the latter group only four isolates required fermentation and assimilation tests to determine whether they were C. parapsilosis (1) or C. guilliermondii (3).


Malawi three district evaluation: Community-based maternal and newborn care economic analysis.
Malawi is one of few low-income countries in sub-Saharan Africa to have met the fourth Millennium Development Goal for child survival (MDG 4). To accelerate progress towards MDGs, the Malawi Ministry of Health's Reproductive Health Unit - in partnership with Save the Children, UNICEF and others - implemented a Community Based Maternal and Newborn Care (CBMNC) package, integrated within the existing community-based system. Multi-purpose Health Surveillance Assistants (HSAs) already employed by the local government were trained to conduct five core home visits. The additional financial costs, including donated items, incurred by the CBMNC package were analysed from the perspective of the provider. The coverage level of HSA home visits (35%) was lower than expected: mothers received an average of 2.8 visits rather than the programme target of five, or the more reasonable target of four given the number of women who would go away from the programme area to deliver. Two were home pregnancy and less than one, postnatal, reflecting greater challenges for the tight time window to achieve postnatal home visits. As a proportion of a 40 hour working week, CBMNC related activities represented an average of 13% of the HSA work week. Modelling for 95% coverage in a population of 100,000, the same number of HSAs could achieve this high coverage and financial programme cost could remain the same. The cost per mother visited would be US$6.6, or US$1.6 per home visit. The financial cost of universal coverage in Malawi would stand at 1.3% of public health expenditure if the programme is rolled out across the country. Higher coverage would increase efficiency of financial investment as well as achieve greater effectiveness.


Microviscosity of the hydrocarbon region of the bovine retinal rod outer segment disk membrane determined by fluorescent probe measurements.
The microviscosity of the hydrocarbon region of the bovine retinal rod outer segment disk membrane was determined by measuring the anisotropy of fluorescence from the probe 1,6-diphenyl-1,3,5-hexatriene. The microviscosity ranged from 1.4 P at 40 degreeC to 15 P at O degree C, and no phase transition was observed in this temperature range. Bleaching of rhodopsin in the disk membrane produced no change in the microviscosity within the limits of error of our measurement (not equal to 5%). The presence of retinal in the disk membrane caused string quenching of the probe fluorescence. Removal of retinal from bleached disks produced a threefold increase in total fluorescence intensity and only a 10% drop in fluorescnece anisotropy, suggesting that large lateral gradients in microviscosity do not exist in the disk membrane. Bilayers prepared from extracted disk lipids had a microviscosity which was about one-fourth that of the intact disk membrane, demonstrating that rhodopsin hinders the mobility of the hydrocarbon chains of the disk phospholipids or egg phosphatidylcholine had identical microviscosities despite the much higher degree of unsaturation of the disk phospholipids.


OBSERVABLE CHANGES OF HYPOTHESES UNDER POSITIVE REINFORCEMENT.
In mathematical models of concept learning it has consistently been assumed that positive reinforcement cannot lead to a change of the hypothesis determining the overt response. When hypotheses are experimentally identified and recorded along with positive and negative reinforcements of stimulus-response pairs, it can be shown that hypotheses may change after a positive reinforcement. Positive reinforcement has an information content for subjects that has not yet been adequately recognized in concept formation studies.


The potent microtubule-stabilizing agent (+)-discodermolide induces apoptosis in human breast carcinoma cells--preliminary comparisons to paclitaxel.
(+)-Discodermolide, a sponge-derived natural product, stabilizes microtubules more potently than paclitaxel despite the lack of any obvious structural similarities between the drugs. It competitively inhibits the binding of paclitaxel to tubulin polymers, hypernucleates microtubule assembly more potently than paclitaxel, and inhibits the growth of paclitaxel-resistant ovarian and colon carcinoma cells. Because paclitaxel shows clinical promise for breast cancer treatment, its effects in a series of human breast cancer cells were compared to those of (+)-discodermolide. Growth inhibition, cell and nuclear morphological, and electrophoretic and flow cytometric analyses were performed on (+)-discodermolide-treated MCF-7 and MDA-MB231 cells. (+)-Discodermolide potently inhibited the growth of both cell types (IC50 < 2.5 nM) at concentrations similar to those observed with paclitaxel. Complete inhibition of growth occurred with 10 nM or greater of each drug and was not reversed by removal. (+)-Discodermolide-treated cells exhibited condensed and highly fragmented nuclei. Flow cytometric comparison of cells treated with either drug at 10 nM, a concentration well below that achieved clinically with paclitaxel, showed both caused cell cycle perturbation and induction of a hypodiploid cell population. (+)-Discodermolide caused these effects more extensively and at earlier time points. The timing and type of high molecular weight DNA fragmentation induced by the two agents was consistent with induction of apoptosis. The results suggest that (+)-discodermolide has promise as a new chemotherapeutic agent against breast and other cancers.


Refinement of a human challenge model for evaluation of enterotoxigenic Escherichia coli vaccines.
Enterotoxigenic Escherichia coli (ETEC) strain H10407 (serotype O78:H11 producing heat-labile toxin [LT], heat-stable toxin [ST], and colonization factor I [CFA/I]) induces reliably high diarrheal attack rates (ARs) in a human challenge model at doses of ≥10(9) CFU. A descending-dose challenge study was conducted with changes to the standard fasting time and buffer formulation, seeking conditions that permit lower inocula while maintaining reproducibly high ARs. In cohort 1, 20 subjects were fasted overnight and randomized 1:1:1:1 to receive H10407 at doses of 10(8) CFU with bicarbonate, 10(8) CFU with CeraVacx, 10(7) CFU with bicarbonate, or 10(7) CFU with CeraVacx. Subsequent cohorts received H10407 (10(7) CFU with bicarbonate) with similar fasting conditions. Cohort 2 included 15 ETEC-naïve volunteers. Cohort 3 included 10 ETEC-naïve volunteers and 10 rechallenged volunteers. In all, 25/35 (71%) ETEC-naïve recipients of 10(7) CFU of H10407 developed moderate or severe diarrhea (average maximum stool output/24 h = 1,042 g), and most (97%) shed H10407 (maximum geometric mean titer = 7.5 × 10(7) CFU/gram of stool). Only one of 10 rechallenged volunteers developed diarrhea. These rechallenged subjects had reduced intestinal colonization, reflected by quantitative microbiology of fecal samples. Among the 35 ETEC-naïve subjects, anti-lipopolysaccharide (LPS) O78 serum antibody responses were striking, with positive IgA and IgG antibody responses in 33/35 (94%) and 25/35 (71%), respectively. Anti-heat-labile enterotoxin (LTB) serum IgA and IgG responses developed in 19/35 (54%) and 14/35 (40%) subjects, respectively. Anti-CFA/I serum IgA and IgG responses were detected in 15/35 (43%) and 8/35 (23%) subjects. After the second challenge, participants exhibited blunted anti-LPS and -LTB responses but a booster response to CFA/I. This ETEC model should prove useful in the future evaluation of ETEC vaccine candidates.


lncRNA ENST00000422059 promotes cell proliferation and inhibits cell apoptosis in breast cancer by regulating the miR-145-5p/KLF5 axis.
Krüppel-like zinc-finger transcription factor 5 (KLF5) is a vital regulator of breast cancer (BC) onset and progression. The mechanism by which KLF5 regulates BC is still not clearly known. In this study, bioinformatics analysis shows that BC-affected individuals with elevated KLF5 expression levels have poor clinical outcomes. We further verify that miR-145-5p regulated KLF5 expression to promote cell apoptosis and inhibit cell proliferation in BC via dual-luciferase reporter assay, western blot analysis, qRT-PCR, CCK-8 assay and cell apoptosis assay. In addition, based on bioinformatics analysis, the binding of ENST00000422059 with miR-145-5p is confirmed by dual-luciferase reporter assay. Subsequently, FISH, western blot analysis, qRT-PCR, CCK-8 and cell apoptosis assays verified that ENST00000422059 increases KLF5 protein expression by sponging miRNA to promote cell proliferation and inhibit cell apoptosis. Finally, ENST00000422059 is found to accelerate tumor progression by regulating the miR-145-5p/KLF5 axis in vivo. In conclusion, this study suggests that ENST00000422059 upregulates KLF5 by sponging miR-145-5p to promote BC progression.


A monoclonal antibody (12G5) directed against CXCR-4 inhibits infection with the dual-tropic human immunodeficiency virus type 1 isolate HIV-1(89.6) but not the T-tropic isolate HIV-1(HxB).
We used a monoclonal antibody (12G5) directed against an extracellular domain of CXCR-4 to investigate the role of this receptor in infection of immortalized lymphoid cell lines, peripheral blood mononuclear cells (PBMCs), and primary brain microglia with a dual-tropic strain of human immunodeficiency virus (HIV-1(89.6)) and a T-tropic strain (HIV-1(IIIB)). Addition of antibody 12G5 to cells prior to and during infection with HIV-1(89.6) inhibited p24 production 100- to 10,000-fold in CEMx174 and 174-CD4 cells and about 10-fold in PBMC cultures but had no activity against infection of either monocyte-derived macrophages or brain microglia. In contrast, 12G5 had little or no effect on infection of CEMx174 cells with HIV-1(IIIB) or HIV-1(HxB). To identify the region of the HIV-1(89.6) envelope that confers sensitivity to 12G5, we used chimeric molecular clones. Chimeras containing the V3 loop region of HIV-1(89.6) were inhibited by 12G5 to the same degree as wild-type HIV-1(89.6) whereas replication of those viruses containing the V3 loop of HIV-1(HxB) was not inhibited by the antibody. A similar pattern was seen in infections of a U87 glioblastoma line that coexpresses CD4 and CXCR-4. Antibody 12G5 was also able to block fusion between HeLa-CD4 cells and CEMx174 cells chronically infected with HIV-1(89.6) but had no effect on fusion mediated by cells chronically infected with HIV-1(IIIB). Taken together, these results suggest that different strains of HIV-1 may interact with different sites on CXCR-4 or may have different binding affinities for the coreceptor.


Mock webpage: an innovative assignment for second-degree nursing students.
Generating challenging and meaningful academic assignments for nursing students in which the student manages the technology used to deliver information is needed for accelerated second-degree nursing students. Developing those assignments is critical for keeping accelerated second-degree students engaged in the learning process. They are highly motivated, self-directed learners who have been found to have a preference for teaching-learning experiences that incorporate the use of technology. This paper describes the development of a mock webpage assignment to meet the learning needs of accelerated second-degree nursing students.


Fv-1 determinants in xenotropic murine leukemia viruses studied with biological assay systems: Isolation of xenotropic virus with N-tropic Fv-1 activity in the cryptic form.
By a biological assay system using phenotypically mixed ecotropic and xenotropic murine leukemia viruses, we investigated whether in the virions of a xenotropic virus there is N- or B-tropic Fv-1 determinant in active form. The existence of N-tropic Fv-1 determinant was demonstrated in SL-XT-1 xenotropic virus isolated from the spleen of a 3-month-old SL mouse, and the N-tropic Fv-1 tropism was confirmed by analysis of the phenotypically mixed viruses harvested from clonal SC-1 cells doubly infected with the SL-XT-1 and B-tropic ecotropic viruses. However, neither N- nor B-tropic Fv-1 determinant was demonstrated in any xenotropic viruses isolated from embryo cells of BALB/c, NZB, or DBA/2 mice, or Cas E #1-IU, and xenotropic-like virus isolated from a wild mouse.


A (Sub)field Guide to Quality Control in Hippocampal Subfield Segmentation on High-resolution T 2 -weighted MRI.
Inquiries into properties of brain structure and function have progressed due to developments in magnetic resonance imaging (MRI). To sustain progress in investigating and quantifying neuroanatomical details in vivo , the reliability and validity of brain measurements are paramount. Quality control (QC) is a set of procedures for mitigating errors and ensuring the validity and reliability of brain measurements. Despite its importance, there is little guidance on best QC practices and reporting procedures. The study of hippocampal subfields in vivo is a critical case for QC because of their small size, inter-dependent boundary definitions, and common artifacts in the MRI data used for subfield measurements. We addressed this gap by surveying the broader scientific community studying hippocampal subfields on their views and approaches to QC. We received responses from 37 investigators spanning 10 countries, covering different career stages, and studying both healthy and pathological development and aging. In this sample, 81% of researchers considered QC to be very important or important, and 19% viewed it as fairly important. Despite this, only 46% of researchers reported on their QC processes in prior publications. In many instances, lack of reporting appeared due to ambiguous guidance on relevant details and guidance for reporting, rather than absence of QC. Here, we provide recommendations for correcting errors to maximize reliability and minimize bias. We also summarize threats to segmentation accuracy, review common QC methods, and make recommendations for best practices and reporting in publications. Implementing the recommended QC practices will collectively improve inferences to the larger population, as well as have implications for clinical practice and public health.


Substance P-mediated expression of the pro-angiogenic factor CCN1 modulates the course of colitis.
Substance P (SP) regulates important intestinal functions, such as mucosal permeability, motility, chloride secretion, and inflammation via the neurokinin-1 receptor (NK-1R). Previous reports showed that vascularization and expression of angiogenic factors are evident in the colonic mucosa of rats with colitis and patients with inflammatory bowel disease. Here we determined whether SP is associated with angiogenesis. Human NCM460 colonocytes stably transfected with the human NK-1R (NCM460-NK-1R cells) and mice with dextran sodium sulfate-induced colitis were used. We found that expression of the angiogenic factor CCN1 was increased in the colons of patients with Crohn's disease and ulcerative colitis. Mucosal extracts from inflammatory bowel disease patients induced human intestinal microvascular endothelial cell migration that was inhibited by blockade of CCN1 and its receptor integrin alphavbeta3. Both the degree of angiogenesis and CCN1 expression were elevated in the colons of mice with dextran sodium sulfate-induced colitis, which was reduced by treatment with the NK-1R antagonist CJ-12255. SP also increased CCN1 expression in NCM460-NK-1R colonocytes. SP exposure to human intestinal microvascular endothelial cells co-cultured with NCM460-NK-1R cells induced angiogenic activity that was inhibited by CCN1 silencing. In addition, intracolonic overexpression of CCN1 induced angiogenesis in mouse colon. Thus, SP mediates angiogenesis via CCN1 during colitis.


Evolution of human calicivirus RNA in vivo: accumulation of mutations in the protruding P2 domain of the capsid leads to structural changes and possibly a new phenotype.
In the present study we report on evolution of calicivirus RNA from a patient with chronic diarrhea (i.e., lasting >2 years) and viral shedding. Partial sequencing of open reading frame 1 (ORF1) from 12 consecutive isolates revealed shedding of a genogroup II virus with relatively few nucleotide changes during a 1-year period. The entire capsid gene (ORF2) was also sequenced from the same isolates and found to contain 1,647 nucleotides encoding a protein of 548 amino acids with similarities to the Arg320 and Mx strains. Comparative sequence analysis of ORF2 revealed 32 amino acid changes during the year. It was notable that the vast majority of the cumulative amino acid changes (8 of 11) appeared within residues 279 to 405 located within the hypervariable domain (P2) of the capsid protein and hence were subject to immune pressure. An interesting and novel observation was that the accumulated amino acid changes in the P2 domain resulted in predicted structural changes, including disappearance of a helix structure, and thus a possible emergence of a new phenotype. FUT2 gene polymorphism characterization revealed that the patient is heterozygous at nucleotide 428 and thus Secretor(+), a finding in accordance with the hypothesis of FUT2 gene polymorphism and calicivirus susceptibility. To our knowledge, this is the first report of RNA evolution of calicivirus in a single individual, and our data suggest an immunity-driven mechanism for viral evolution. We also report on chronic virus excretion, immunoglobulin treatment, and modification of clinical symptoms; our observations from these studies, together with the FUT2 gene characterization, may lead to a better understanding of calicivirus pathogenesis.


Short-term augmentation of venous drainage with extra-corporeal shunt and simultaneous auto-transfusion, for salvaging a congested free flap.
Adequate drainage of venous blood is the most critical part of successful free tissue transfer. We report a case of anterolateral thigh flap used for covering open communited tibial fracture. The flap was salvaged with short term augmentation of venous drainage with external shunt. The drainage was continued for six days. It was confirmed that there is no more congestion after blocking the catheter and then the drainage was discontinued on seventh day. The flap was successfully salvaged. This method has potential applications in multiple situations for successful salvage of free tissue transfer.


Single-cell analysis reveals individual spore responses to simulated space vacuum.
Outer space is a challenging environment for all forms of life, and dormant spores of bacteria have been frequently used to study the survival of terrestrial life in a space journey. Previous work showed that outer space vacuum alone can kill bacterial spores. However, the responses and mechanisms of resistance of individual spores to space vacuum are unclear. Here, we examined spores' molecular changes under simulated space vacuum (~10-5 Pa) using micro-Raman spectroscopy and found that this vacuum did not cause significant denaturation of spore protein. Then, live-cell microscopy was developed to investigate the temporal events during germination, outgrowth, and growth of individual Bacillus spores. The results showed that after exposure to simulated space vacuum for 10 days, viability of spores of two Bacillus species was reduced up to 35%, but all spores retained their large Ca2+-dipicolinic acid depot. Some of the killed spores did not germinate, and the remaining germinated but did not proceed to vegetative growth. The vacuum treatment slowed spore germination, and changed average times of all major germination events. In addition, viable vacuum-treated spores exhibited much greater sensitivity than untreated spores to dry heat and hyperosmotic stress. Among spores' resistance mechanisms to high vacuum, DNA-protective α/β-type small acid-soluble proteins, and non-homologous end joining and base excision repair of DNA played the most important roles, especially against multiple cycles of vacuum treatment. Overall, these results give new insight into individual spore's responses to space vacuum and provide new techniques for microorganism analysis at the single-cell level.


Carboxy-terminal analysis of poliovirus proteins: termination of poliovirus RNA translation and location of unique poliovirus polyprotein cleavage sites.
The carboxy-terminal amino acids of a number of poliovirus proteins were determined by carboxypeptidase A analysis. The nonstructural proteins P3-2, P3-4b and their precursor. P3-1b, were found to be coterminal with a sequence of -Ser-Phe-COOH. As these proteins are coded for at the extreme 3' end of the viral RNA, it is possible to establish the termination site of translation at nucleotide 7,361, 73 nucleotides before the start of the polyadenylic acid tract of the RNA. Two additional nonstructural proteins, P2-X and its precursor, P2-3b, were also found to be coterminal with a sequence of -Phe-Gln-COOH. This result confirms the existence of at least one Gln-Gly proteolytic cleavage site. These Gln-Gly cleavage sites are predicted from the nucleotide sequence to be ubiquitous throughout the poliovirus genome. The only exceptions are the cleavage sites at the carboxy termini of the structural protein VP4 and VP1. Carboxypeptidase A analysis of VP1 establishes a terminal sequence of -Thr-Tyr-COOH, and similar analysis of VP4 shows Asn to be the terminal amino acid residue, observations that prove the existence of the exceptional C-terminal amino acids. In none of the analyzed cases has C-terminal trimming after cleavage been observed.


Pain and the diagnostic process: conceptions and misconceptions.
Pain may be considered a diagnostic alarm that acknowledges but usually fails to determine the etiology of a particular clinical problem. Diagnosis thus involves the extraction of additional information from a patient in order to determine rational bases for treatment. The process is ill-defined and fraught with error unless the diagnostician is aware of all and only the relevant diagnostic alternatives. This list of alternatives plus an appreciation of the likelihood of each possible diagnostic outcome determines the fastest rate that diagnostic information can be processed by any system in the absence of unpredictable events called noise. Human diagnosticians must be fluent in the "language" used to convey the desired information if performance is to be optimized. In general this involves restructuring the diagnostic task into smaller pieces which are easier to digest perceptually. One way to accomplish this is to ignore or eliminate patterns which are either irrelevant or occur so rarely (or so commonly) that they usually have negligible impact on diagnostic performance. The utility of a diagnosis is determined ultimately by the degree to which it influences the health and well-being of both the patient and society at large. Hence, the consequence of a particular disease may offset the low diagnostic yield per examination resulting from a low or high prevalence of the disease. When diagnostic patterns are limited by ignorance to only signs and/or symptoms such as pain, the diagnostic process loses its meaning and treatment decisions are compromised necessarily by uncertainty.


Quantitative multiphoton microscopy of murine urinary bladder morphology during in situ uniaxial loading.
Urodynamic tests are the gold standard for the diagnosis of bladder dysfunction, and the mechanical compliance of the bladder is an important parameter in these tests. The bladder wall has a layered structure, differentially affected by pathology, so knowledge of the contribution and role of these layers and their constituents to overall bladder compliance will enhance interpretation of these clinical tests. In this study we document the functional morphology of the detrusor and lamina propria of the murine bladder wall using a custom in-situ tensile loading system under multiphoton microscopy (MPM) observation in unloaded state and under incremental uniaxial stretch. Features in the stress-stretch curves of bladder samples were then directly related to corresponding MPM images. Collagen organisation across wall depth was quantified using image analysis techniques. The hypothesis that the lamina propria deformed at low strain by unfolding of the rugae and rearranging collagen fibrils was confirmed. A novel 'pocket' feature in the detrusor was observed along with extensive rearrangement of fibrils in two families at different depths, providing higher stiffness at high stretches in the detrusor. The very different deformations of detrusor and lamina propria were accommodated by the highly coiled structure of collagen in the lamina propria. Imaging and mechanical studies presented here allow gross mechanical response to be attributed to specific components of the bladder wall and further, may be used to investigate the impact of microstructural changes due to pathology or aging, and how they impair tissue functionality. This article reports the first in-situ multiphoton microscopy observations of microstructural deformation under uniaxial tensile loading of ex vivo bladder. We describe collagen rearrangement through the tissue thickness and relate this directly to the stress-stretch behaviour. We confirm for the first time the unfolding of rugae and realignment of fibrils in the lamina propria during extension and the rapid stiffening as two fibril families in the detrusor are engaged. This technique provides new insight into microstructure function and will enhance understanding of the impact of changes due to pathology or aging.


Genomic selection identifies vertebrate transcription factor Fezf2 binding sites and target genes.
Identification of transcription factor targets is critical to understanding gene regulatory networks. Here, we uncover transcription factor binding sites and target genes employing systematic evolution of ligands by exponential enrichment (SELEX). Instead of selecting randomly synthesized DNA oligonucleotides as in most SELEX studies, we utilized zebrafish genomic DNA to isolate fragments bound by Fezf2, an evolutionarily conserved gene critical for vertebrate forebrain development. This is, to our knowledge, the first time that SELEX is applied to a vertebrate genome. Computational analysis of bound genomic fragments predicted a core consensus binding site, which identified response elements that mediated Fezf2-dependent transcription both in vitro and in vivo. Fezf2-bound fragments were enriched for conserved sequences. Surprisingly, ∼20% of these fragments overlapped well annotated protein-coding exons. Through loss of function, gain of function, and chromatin immunoprecipitation, we further identified and validated eomesa/tbr2 and lhx2b as biologically relevant target genes of Fezf2. Mutations in eomesa/tbr2 cause microcephaly in humans, whereas lhx2b is a critical regulator of cell fate and axonal targeting in the developing forebrain. These results demonstrate the feasibility of employing genomic SELEX to identify vertebrate transcription factor binding sites and target genes and reveal Fezf2 as a transcription activator and a candidate for evaluation in human microcephaly.


Prevention and treatment of urinary incontinence after stroke in adults.
Urinary incontinence can affect 40-60% of people admitted to hospital after a stroke, with 25% still having problems on hospital discharge and around 15% remaining incontinent at one year. To determine the optimal methods for prevention and treatment of urinary incontinence after stroke in adults. We searched the Cochrane Incontinence and Stroke Groups specialised registers (searched 15 December 2004 and 26 October 2004, respectively), CINAHL (January 1982 to November 2004), national and international trial databases for unpublished data, and the reference lists of relevant articles. Randomised or quasi-randomised controlled trials evaluating the effects of interventions designed to promote continence in people after stroke. Data extraction and quality assessment were undertaken by two reviewers working independently. Disagreements were resolved by a third reviewer. Seven trials with a total of 399 participants were included in the review. Participants were from a mixture of settings, age groups and phases of stroke recovery. No two trials addressed the same comparison. Four trials tested an intervention against usual care, including acupuncture, timed voiding, and two types of specialist professional intervention. One cross-over trial tested an intervention (oestrogen) against placebo. One trial tested a specific intervention (oxybutynin) against another intervention (timed voiding), and one trial tested a combined intervention (sensory-motor biofeedback plus timed voiding) against a single component intervention (timed voiding alone). Reported data were insufficient to evaluate acupuncture or timed voiding versus usual care, oxybutynin versus timed voiding, or sensory motor biofeedback plus timed voiding versus usual care. Evidence from a single small trial suggested that structured assessment and management of care in early rehabilitation may reduce the number of people with incontinence at hospital discharge (1/21 versus 10/13; RR 0.06, 95% CI 0.01 to 0.43), and have other benefits. Evidence from another trial suggested that assessment and management of care by Continence Nurse Practitioners in a community setting may reduce the number of urinary symptoms (48/89 versus 38/54; RR 0.77, 95% CI 0.59 to 0.99), and increase satisfaction with care. There was suggestive evidence that specialist professional input through structured assessment and management of care and specialist continence nursing may reduce urinary incontinence after stroke. Data from trials of other physical, behavioural, complementary and anticholinergic drug interventions are insufficient to guide continence care of adults after stroke.


Response of bone marrow stromal cells to graded co-electrospun scaffolds and its implications for engineering the ligament-bone interface.
Biomaterial scaffolds with gradients in architecture, mechanical and chemical properties have the potential to improve the osseointegration of ligament grafts by recapitulating phenotypic gradients that exist at the natural ligament-bone (L-B) interface. Towards the larger goal of regenerating the L-B interface, this in vitro study was performed to investigate the potential of two scaffolds with mineral gradients in promoting a spatial gradient of osteoblastic differentiation. Specifically, the first graded scaffold was fabricated by co-electrospinning two polymer solutions (one doped with nano-hydroxyapatite particles) from offset spinnerets, while the second was created by immersing the first scaffold in a 5 × simulated body fluid. Rat bone marrow stromal cells, cultured in the presence of osteogenic supplements, were found to be metabolically active on all regions of both scaffolds after 1 and 7 days of culture. Gene expression of bone morphogenic protein-2 and osteopontin was elevated on mineral-containing regions as compared to regions without mineral, while the expression of alkaline phosphatase mRNA revealed the opposite trend. Finally, the presence of osteopontin and bone sialoprotein confirmed osteoblastic phenotypic maturation by day 28. This study indicates that co-electrospun scaffolds with gradients in mineral content can guide the formation of phenotypic gradients and may thus promote the regeneration of the L-B interface.