Adult brain activation in response to pain is changed by neonatal painful stimulation according to sex: A manganese-enhanced MRI study.

Although it is known that nociceptive stimulation in the first postnatal week in rats is useful to model preterm pain, resulting in activation of specific brain areas, as assessed in vivo using manganese-enhanced magnetic resonance imaging (MEMRI), little is known about its long-term effects and sex specificity. Here we aimed to investigate whether inflammatory pain induced in male and female adult rats modify the pattern of brain activation between animals subjected or not to neonatal pain. For this, Complete Freund's adjuvant (CFA) was injected into the left hind paw of rat pups on postnatal day 1 (P1) or P8 to induce inflammatory response. During adulthood, CFA-treated and control animals were injected with CFA 1 hr prior MRI. MEMRI has the ability to enhance the contrast of selective brain structures in response to a specific stimulus, as the pain. MEMRI responses were consistent with activation of nociceptive pathways and these responses were reduced in animals treated with CFA on P1, but increased in animals treated on P8, mainly in the female group. In agreement, P8 female group showed exacerbated responses in the thermal nociceptive test. Using MEMRI, we conclude that the natural ability of adult rats to recognize and react to pain exposition is modified by neonatal painful exposition, mainly among females.

Changes in Hippocampal Volume are Correlated with Cell Loss but Not with Seizure Frequency in Two Chronic Models of Temporal Lobe Epilepsy.

Kainic acid (KA) or pilocarpine (PILO) have been used in rats to model human temporal lobe epilepsy (TLE) but the distribution and severity of structural lesions between these two models may differ. Magnetic resonance imaging (MRI) studies have used quantitative measurements of hippocampal T2 (T2HP) relaxation time and volume, but simultaneous comparative results have not been reported yet. The aim of this study was to compare the MRI T2HP and volume with histological data and frequency of seizures in both models. KA- and PILO-treated rats were imaged with a 2 T MRI scanner. T2HP and volume values were correlated with the number of cells, mossy fiber sprouting, and spontaneous recurrent seizures (SRS) frequency over the 9 months following status epilepticus (SE). Compared to controls, KA-treated rats had unaltered T2HP, pronounced reduction in hippocampal volume and concomitant cell reduction in granule cell layer, CA1 and CA3 at 3 months post SE. In contrast, hippocampal volume was unchanged in PILO-treated animals despite detectable increased T2HP and cell loss in granule cell layer, CA1 and CA3. In the following 6 months, MRI hippocampal volume remained stable with increase of T2HP signal in the KA-treated group. The number of CA1 and CA3 cells was smaller than age-matched CTL group. In contrast, PILO group had MRI volumetric reduction accompanied by reduction in the number of CA1 and CA3 cells. In this group, T2HP signal was unaltered at 6 or 9 months after status. Reductions in the number of cells were not progressive in both models. Notably, the SRS frequency was higher in PILO than in the KA model. The volumetry data correlated well with tissue damage in the epileptic brain, suggesting that MRI may be useful for tracking longitudinal hippocampal changes, allowing the assessment of individual variability and disease progression. Our results indicate that the temporal changes in hippocampal morphology are distinct for both models of TLE and that these are not significantly correlated to the frequency of SRS.

First Report of Downy Mildew on Blackberry Caused by Peronospora sparsa in Arkansas.

In March of 2014, blackberry (Rubus fruticosus) greenhouse seedlings with leaf symptoms that included yellowing on the leaf surface and reddish brown angular lesions with necrotic centers and chlorotic margins were detected in the University of Arkansas Fruit Research Station in Clarksville, AR (35°32.065' N, 93°24.3564' W). Symptoms were observed on multiple blackberries in the greenhouse, with an estimated prevalence of 25%. Three symptomatic samples of the affected plants were submitted to the Plant Health Clinic in Fayetteville, AR, for diagnosis. Sporulation was observed on the underside of the symptomatic leaf tissue. Hyaline sporangiophores were observed emerging from stomata on the undersides of leaves, 295 to 620 × 4 to 6 µm with long, straight trunks and were branched 3 to 4 times with bifurcated tips, with a length of 5 to 23 µm. Typically, one branch of each pair curved inward and one reflexed. Sporangiophores ended with sporangia that were round or slightly ovoid, colorless to yellowish-brown, and 14 to 22 × 11 to 18 µm. The causal agent was morphologically identified as Peronospora sparsa Berk (1,2). The identification was confirmed using a molecular method directly from plant tissue. DNA was extracted from two samples (~3 × 3 mm) from each of three symptomatic leaves, followed by PCR amplification using P. sparsa-specific rDNA-ITS region primers P1: 5'-CACGTGAACCGTATCAACC-3' and P2: 5'-GATAGGGCTTGCCCAGTAG-3' (GenBank Accession No. Y15816) (4). DNA amplification was successful, resulting in a product of 94 bp, confirming that P. sparsa was present in the symptomatic blackberry tissue. Sporulating leaf tissue was laid on the underside of surface sterilized blackberry leaves from three plants with a similar genetic background and incubated at 17°C with a 12-h photoperiod in a moist chamber. Sporangiophores and sporangia developed on the underside of lesions on previously uninfected leaves 16 days after inoculation. As a control, leaves from the same three plants were surface sterilized and placed in a moist chamber and incubated at 17°C with a 12-h photoperiod and examined 16 days later. No lesions or sporulation developed on the controls. Previously, P. sparsa has been reported on blackberry in California (3) and Mexico (5). To our knowledge, this is the first report of P. sparsa causing downy mildew on blackberry in Arkansas. References: (1) M. J. Berkeley. Gardeners' Chronicle 14:307, 1861. (2) M. A. Ellis. Page 15 in: Compendium of Raspberry and Blackberry Diseases and Insects. APS Press, St. Paul, MN, 1989. (3) D. Farr et al. Page 486 in: Fungi on Plants and Plant Products in the United States. APS Press, St. Paul, MN, 1989. (4) A. Hukkanen et al. Eur. J. Plant Pathol. 116:225, 2006. (5) A. Rebollar-Alviter et al. Plant Dis. 93:674, 2009.

Manganese-enhanced magnetic resonance imaging detects mossy fiber sprouting in the pilocarpine model of epilepsy.

PURPOSE: Mossy fiber sprouting (MFS) is a frequent finding following status epilepticus (SE). The present study aimed to test the feasibility of using manganese-enhanced magnetic resonance imaging (MEMRI) to detect MFS in the chronic phase of the well-established pilocarpine (Pilo) rat model of temporal lobe epilepsy (TLE). METHODS: To modulate MFS, cycloheximide (CHX), a protein synthesis inhibitor, was coadministered with Pilo in a subgroup of animals. In vivo MEMRI was performed 3 months after induction of SE and compared to the neo-Timm histologic labeling of zinc mossy fiber terminals in the dentate gyrus (DG). KEY FINDINGS: Chronically epileptic rats displaying MFS as detected by neo-Timm histology had a hyperintense MEMRI signal in the DG, whereas chronically epileptic animals that did not display MFS had minimal MEMRI signal enhancement compared to nonepileptic control animals. A strong correlation (r = 0.81, p < 0.001) was found between MEMRI signal enhancement and MFS. SIGNIFICANCE: This study shows that MEMRI is an attractive noninvasive method for detection of mossy fiber sprouting in vivo and can be used as an evaluation tool in testing therapeutic approaches to manage chronic epilepsy.

LY2439821, a humanized anti-interleukin-17 monoclonal antibody, in the treatment of patients with rheumatoid arthritis: A phase I randomized, double-blind, placebo-controlled, proof-of-concept study.

OBJECTIVE: We undertook this study to evaluate safety, tolerability, pharmacokinetics, pharmacodynamics, and efficacy of LY2439821, a humanized anti-interleukin-17 (anti-IL-17) monoclonal antibody, in a first in-human trial in rheumatoid arthritis (RA) patients taking oral disease-modifying antirheumatic drugs (DMARDs). METHODS: This randomized, double-blind, placebo-controlled study consisted of 2 parts. In part A, 20 patients received 1 intravenous (IV) dose of LY2439821 (0.06, 0.2, 0.6, or 2.0 mg/kg, escalating) or placebo followed by 8 weeks of evaluation. End points included safety, tolerability, and pharmacokinetics. In part B, 77 patients received 1 IV dose of LY2439821 (0.2, 0.6, or 2.0 mg/kg) or placebo every 2 weeks for a total of 5 doses, with a total evaluation period of 16 weeks. End points included safety, tolerability, pharmacokinetics/pharmacodynamics, and efficacy (Disease Activity Score in 28 joints [DAS28] and percentages of patients meeting American College of Rheumatology 20%, 50%, or 70% improvement criteria [achieving an ACR20, ACR50, or ACR70 response]). The primary efficacy end point was the DAS28 at week 10. RESULTS: Baseline characteristics were similar across all groups. Changes in the DAS28 were significantly greater in the 0.2 mg/kg, 2.0 mg/kg, and all-LY2439821-combined groups (-2.3, -2.4, and -2.3, respectively) than in the placebo group (-1.7) at week 10 (P < or = 0.05), and these differences were significant as early as week 1. Percentages of ACR20, ACR50, and ACR70 responses as well as improvements in the ACR core set of measures were greater in LY2439821-treated patients than in placebo-treated patients at multiple time points. There was no apparent dose-response relationship in treatment-emergent adverse events. CONCLUSION: LY2439821 added to oral DMARDs improved signs and symptoms of RA, with no strong adverse safety signal noted. This first evaluation of LY2439821 supports neutralization of IL-17 as a potential novel goal for the treatment of RA.

Automated extraction of direct, reactive, and vat dyes from cellulosic fibers for forensic analysis by capillary electrophoresis.

Systematic designed experiments were employed to find the optimum conditions for extraction of direct, reactive, and vat dyes from cotton fibers prior to forensic characterization. Automated microextractions were coupled with measurements of extraction efficiencies on a microplate reader UV-visible spectrophotometer to enable rapid screening of extraction efficiency as a function of solvent composition. Solvent extraction conditions were also developed to be compatible with subsequent forensic characterization of extracted dyes by capillary electrophoresis with UV-visible diode array detection. The capillary electrophoresis electrolyte successfully used in this work consists of 5 mM ammonium acetate in 40:60 acetonitrile-water at pH 9.3, with the addition of sodium dithionite reducing agent to facilitate analysis of vat dyes. The ultimate goal of these research efforts is enhanced discrimination of trace fiber evidence by analysis of extracted dyes.

Neutral particle analyzer measurements on the SSPX spheromak.

A neutral particle analyzer is used to measure the time-resolved energy spectrum of neutral hydrogen leaving a spheromak plasma. A gas cell filled with 10-50 mTorr of helium is used to strip electrons from incoming neutral hydrogen, lowering the minimum detectable energy well below that obtained with thin foils. Effective neutral particle temperature is calculated by fitting a Maxwellian energy distribution to the measured energy spectrum above and below approximately 300 eV. A computational model with approximated profiles of plasma density and neutral density is used with the measured neutral hydrogen flux to estimate the ion temperature. Measurement of the power flux due to neutral hydrogen emitted at the measurement location is extended to the whole plasma surface to estimate the total charge exchange power loss from the plasma. The initial results indicate that the charge exchange power loss represents only 2% of the total input gun power during the sustainment phase of the discharge.

Preparation and Certification of K-411 Glass Microspheres.

The production and characterization of NBS K-411 glass microspheres in the 2-40 µm range for certification as NIST Standard Reference Material(R) 2066 (SRM(R)) are described. Quantitative analysis and heterogeneity testing of the microspheres were done with an electron probe microanalyzer-X-ray energy dispersive spectrometry (EPMA-EDS) automated particle analysis procedure. Results for the trimmed and normalized data produced mean compositions for the elements Mg, Si, Ca, Fe, and O (calculated from stoichiometry) that are in good agreement with the certified values for the K-411 bulk glass (NBS SRM 470 Glasses for Mineral Analysis), but with uncertainties about twice as large as those for the bulk material. Differences from the bulk are attributable to microsphere geometry as well as mass and size effects.

Elemental and molecular imaging of human hair using secondary ion mass spectrometry.

Secondary ion mass spectrometry (SIMS) is used to image the spatial distribution of elemental and molecular species on the surface and in cross sections of doped human hair using a magnetic sector SIMS instrument operated as an ion microprobe. Analysis of electrically insulating, non-planar hair samples requires one of two different methods of charge compensation to be used depending on the polarity of the sputtered secondary ions. For detection of positive secondary ions, the hair is imaged using a approximately 0.5 micron diameter, 19.5 keV impact energy, O- microbeam with no auxiliary electron bombardment. For detection of negative secondary ions, a approximately 0.2 micron diameter, 14.5 keV impact energy Cs+ microbeam is used in conjunction with normal incidence, low-energy electron bombardment. Both of these methods allow submicrometer spatial resolution elemental and molecular secondary ion images to be obtained from hair samples without metallic coating of the sample surface prior to analysis. Several examples are presented that reflect potential application areas for these analytical methods.

Mometasone furoate nasal spray is rapidly effective in the treatment of seasonal allergic rhinitis in an outdoor (park), acute exposure setting.

The objective of this study was to determine the time to onset of symptom relief following a single dose of mometasone furoate nasal spray (MFNS) in symptomatic patients with seasonal allergic rhinitis (SAR). This was a single-center, placebo-controlled, double-blind, randomized, parallel-group study with a 7-day run-in period followed by a single-dose administration of medication or placebo in an outdoor park setting. The park site provided an acute exposure to seasonal (tree and grass) pollens. Patients remained in the park of approximately 12 hours after dosing, during which time hourly assessments of SAR symptoms were recorded on a diary card. Two hundred thirty-nine patients with symptoms of SAR entered the study. Patients receiving any concurrent medication for treatment of their symptoms were excluded. Patients were randomized in a 1:1 ratio to receive treatment with either a single dose of MFNS (200 micrograms/or matching placebo nasal spray. Outcome measures included an assessment of overall therapeutic response and change from baseline in total nasal plus non-nasal sign/symptom severity score, total nasal sign/symptom severity score, and total non-nasal sign/symptom severity score. Improvement in total nasal symptom scores, total non-nasal symptom scores, and total nasal plus non-nasal symptom scores were greater and more sustained in patients receiving MFNS than in patients receiving placebo. The mean decrease from baseline in total nasal plus non-nasal symptom scores was significantly greater in MFNS-dosed patients than in placebo-dosed patients at 5 hours after dosing (p < 0.01). The mean decrease from baseline in total nasal symptom scores was significantly greater in MFNS-dosed patients than in placebo-dosed patients at 7 hours after dosing (p < 0.01). The between-treatment differences in total nasal plus non-nasal symptom scores and total nasal symptom scores remained significant for all subsequent hourly assessments through 12 hours post-dose. Patient assessments of overall response to therapy at end point were significantly different between treatment groups (p < 0.01) with 60.5% of MFNS-treated patients reporting complete, marked, or moderate relief compared with 46.5% of placebo-treated patients. Mometasone furoate nasal spray produces a statistically significant improvement in nasal symptom scores in patients with SAR by 7 hours after administration of a single 200 micrograms dose (100 micrograms in each nostril).

Preliminary evaluation of an SF5+ polyatomic primary ion beam for analysis of organic thin films by secondary ion mass spectrometry.

Organic vapor deposited thin films of pure biomolecules, polymer films and biomolecules dispersed in gelatin and biological tissue have been analyzed in a magnetic sector secondary ion mass spectrometer using an SF5+ primary ion beam at keV impact energies. In comparison to Ar+ bombardment under identical conditions, bombardment with SF5+ gives a 10 to 50 fold enhancement in the secondary ion yields for characteristic molecular ions. The SF5+ primary ion beam can be focussed to a small spot allowing molecular ion images to be obtained at micrometer spatial resolution with enhanced sensitivity. More importantly, the decay in molecular ion signal as a function of primary ion dose commonly observed in SIMS using monoatomic primary ions is either eliminated or greatly reduced, allowing molecular depth profiles to be obtained of organic thin films. By continuing to sample intact molecules as sputtering proceeds into the sample, the total number of detected characteristic secondary ions is increased by as much as a factor of approximately 700 for SF5+ bombardment as compared to Ar+ bombardment under identical analytical conditions. This effect is thought to be a result of the high erosion rate and the low penetration depth inherent in the use of a polyatomic primary projectile.

Effect of Donjoy Ankle Ligament Protector and Aircast Sport-Stirrup orthoses on functional performance.

Clinicians often are faced with the task of selecting an ankle orthosis that will provide protection against ankle sprain injury and will not have a deleterious effect on functional performance. The purpose of this study was to compare the effects of the DonJoy Ankle Ligament Protector (ALP) and the Aircast Sport-Stirrup (AS) on three functional performance tasks: the 40-m sprint run, the figure-of-eight run, and the standing vertical jump. Subjects were eight males and eight females who reported no history of ankle injury during the 6 months prior to testing; neurological condition; lower extremity surgery or pathology; or cardiac, pulmonary, vascular, or balance problems. Each subject performed all functional tasks on the first test day with both ankles unbraced and with one randomly selected ankle braced with one of the two orthoses. Each subject returned for testing on a second day and performed all tasks with both ankles unbranced and with the other orthosis on the previously selected ankle. Subjects also rated the orthoses for comfort and support. An ANOVA indicated that braced data were not significantly different than unbraced data and that data for the two orthoses did not differ significantly for any of the functional performance tasks. Seventy-five percent of the subjects reported the AS was more comfortable than the ALP, and 63% of the subjects indicated that the ALP provided more support than the AS. Neither orthosis had a deleterious effect on functional performance. The results may assist clinicians in selecting either of these orthoses for use in protection against ankle sprain injury.

Rectal carcinoma. Long-term experience with moderately high-dose preoperative radiation and low anterior resection.

This report updated an analysis of a 14-year experience of moderately high-dose (4500 to 5000 cGy) preoperative radiation as an adjuvant to low anterior resection of 95 cases of adenocarcinoma of the rectum. The treatment was well tolerated without treatment-related mortality and with a low incidence (5.2%) of severe complications. The local recurrence rate was 4.2%, and distant failure rate was 10.5%. At 5 years, the actuarial survival rate was 66% and the disease-free survival rate was 64%. At 10 years, the actuarial survival rate and disease-free survival rate were 52%. The authors concluded that moderately high-dose (4500 to 5000 cGy) neoadjuvant radiation in clinically resectable adenocarcinoma of the rectum in which one segment of the anastomosis was in the preoperative radiation field is a safe, effective adjuvant to low anterior resection and that it offered patients excellent local control, long-term survival, and sphincter preservation. Results could be enhanced by chemotherapy, and the authors urged well-designed prospective randomized multicenter trials to determine the most appropriate drugs, dosage, and sequencing of co-adjuvant preoperative radiation therapy and chemotherapy with surgery.

Characterization of effector functions of v-raf/mil- and v-myc-transformed murine splenic macrophage cell lines.

We have characterized several of the cytocidal effector functions of a series of cell lines derived by recombinant retroviral transformation of individual clones of C3H/HeJ mouse splenic macrophages. The three cell lines described in this report (4.01, 4.07, 4.14) all expressed equivalent tumoricidal activity against P815 tumor target cells. However they differed in their high avidity binding of tumor cells (4.01 = 4.14 greater than 4.07), as well as in the killing of Leishmania major (4.01 = 4.07 greater than 4.14), the expression of antibody-dependent cellular cytotoxicity against chicken erythrocytes (4.14 greater than 4.01 greater than 4.07), and finally, in the tumor-stimulated release of tumor necrosis factor-alpha (4.01 = 4.14 greater than 4.07). The stable and restricted expression of distinct effector functions among these three cell lines makes them particularly valuable as models for establishing the precise mechanisms by which cytocidal functions are effected. In addition, they should also prove of value in understanding the basis for macrophage functional diversity.

Electron microscopic evaluation of bacterial adherence to polyvinyl chloride endotracheal tubes used in neonates.

Studies in adults suggest a possible association between pulmonary nosocomial infection and bacterial adherence to endotracheal tubes (ETT). Some strains of coagulase-negative Staphylococcus produce a mucoid extracellular material known as slime or glycocalyx that helps bacteria adhere to the surface of biomaterials. We examined by scanning and transmission electron microscope the surface of 29 polyvinyl ETT removed from neonatal patients, and ten unused ETT. Multiple surface irregularities were found in the unused tubes. These irregularities were most prominent in the tip and around the side hole. Amorphous material containing slime-producing bacteria was found on 29 ETT removed from patients. Tubes in place for longer times had greater areas of surface covered; the amorphous material was more concentrated in the areas where surface irregularities were most prominent. No clinical correlation with pulmonary nosocomial infection was established.

Immortalization of cloned mouse splenic macrophages with a retrovirus containing the v-raf/mil and v-myc oncogenes.

The recombinant retrovirus J2, which contains the v-raf/mil and v-myc oncogenes, was used to immortalize mouse splenic macrophages that had been cloned in soft agar. When added to freshly harvested colonies, J2 failed to yield cell lines but it immortalized up to 30% of the clones if they had been maintained for at least 4 months in medium containing colony-stimulating factor 1 (CSF-1). All of the cell lines grew in agar in a CSF-1-independent manner, and they produced tumors in nude and syngeneic mice. The cell lines were judged to be macrophage based on morphological criteria and because they secreted lysozyme, were phagocytic for antibody-coated particles, and expressed both the Mac-1 antigen and the CSF-1 receptor. The cell lines could be divided into three groups based on their expression of Ia and their ability to present an antigen to a T-cell hybridoma. The majority of the lines did not constitutively express Ia or present antigen, but a lymphokine did induce Ia in all of the lines, with most of them also acquiring antigen-presenting activity. However, a small proportion of lymphokine-treated lines continued to lack antigen-presenting activity despite their ability to express Ia. The third and smallest group of cell lines constitutively expressed both Ia and antigen-presenting activity. These results show that the J2 recombinant retrovirus is a useful means of immortalizing functionally distinct populations of cloned splenic macrophages.

Is anterior resection of the rectosigmoid safe after preoperative radiation?

Forty patients with carcinoma of the rectum or rectosigmoid underwent preoperative irradiation, followed by anterior resection with anastomosis. The radiation dose was 4500 rads administered in 25 fractions over a period of five weeks. One to three weeks after the radiation was completed, anterior resection of the rectosigmoid with anastomosis was done. Of the 40 patients, 23 had hand-sewn anastomoses and 17 had EEA stapled anastomoses. Fourteen of the 40 had diverting loop colostomies, all colostomies were subsequently closed, and there were no clinical postoperative anastomotic leaks. Anterior resection and anastomosis are considered to be technically safe with the surgical and radiation techniques used.

A physical map of caprine arthritis-encephalitis provirus.

Terminally redundant linear proviral DNA of approximately 9.5 kb was the major unintegrated species recovered in the Hirt supernatant fraction of caprine synovial membrane cells infected with strain 75-G63 caprine arthritis-encephalitis virus. A physical map based on the cleavage sites of 13 restriction endonucleases was deduced for this proviral DNA.