Brain glucose hypometabolism and hippocampal inflammation in Goto-Kakizaki rats.

Brain glucose hypometabolism and neuroinflammation are early pathogenic manifestations in neurological disorders. Neuroinflammation may also disrupt leptin signaling, an adipokine that centrally regulates appetite and energy balance by acting on the hypothalamus and exerting neuroprotection in the hippocampus. The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus (T2DM) animal model used to investigate diabetes-associated molecular mechanisms without obesity jeopardizing effects. Wistar and GK rats received the maintenance adult rodent diet. Also, an additional control group of Wistar rats received a high-fat and high-sugar diet (HFHS) provided by free consumption of condensed milk. All diets and water were provided ad libitum for eight weeks. Brain glucose uptake was evaluated by 2-deoxy-2-[fluorine-18] fluoro-D-glucose under basal (saline administration) or stimulated (CL316,243, a selective ß3-AR agonist) conditions. The animals were fasted for 10-12 h, anesthetized, and euthanized. The brain was quickly dissected, and the hippocampal area was sectioned and stored at -80°C in different tubes for protein and RNA analyses on the same animal. GK rats exhibited attenuated brain glucose uptake compared to Wistar animals and the HFHS group under basal conditions. Also, the hippocampus of GK rats displayed upregulated leptin receptor, IL-1ß, and IL-6 gene expression and IL-1ß and the subunit of the transcription factor NF-κB (p-p65) protein expression. No significant alterations were detected in the hippocampus of HFHS rats. Our data indicated that a genetic predisposition to T2DM has significant brain deteriorating features, including brain glucose hypometabolism, neuroinflammation, and leptin signaling disruption in the hippocampal area.

Brain glucose hypometabolism and hippocampal inflammation in Goto-Kakizaki rats

Brain glucose hypometabolism and neuroinflammation are early pathogenic manifestations in neurological disorders. Neuroinflammation may also disrupt leptin signaling, an adipokine that centrally regulates appetite and energy balance by acting on the hypothalamus and exerting neuroprotection in the hippocampus. The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus (T2DM) animal model used to investigate diabetes-associated molecular mechanisms without obesity jeopardizing effects. Wistar and GK rats received the maintenance adult rodent diet. Also, an additional control group of Wistar rats received a high-fat and high-sugar diet (HFHS) provided by free consumption of condensed milk. All diets and water were provided ad libitum for eight weeks. Brain glucose uptake was evaluated by 2-deoxy-2-[fluorine-18] fluoro-D-glucose under basal (saline administration) or stimulated (CL316,243, a selective β3-AR agonist) conditions. The animals were fasted for 10-12 h, anesthetized, and euthanized. The brain was quickly dissected, and the hippocampal area was sectioned and stored at -80°C in different tubes for protein and RNA analyses on the same animal. GK rats exhibited attenuated brain glucose uptake compared to Wistar animals and the HFHS group under basal conditions. Also, the hippocampus of GK rats displayed upregulated leptin receptor, IL-1β, and IL-6 gene expression and IL-1β and the subunit of the transcription factor NF-κB (p-p65) protein expression. No significant alterations were detected in the hippocampus of HFHS rats. Our data indicated that a genetic predisposition to T2DM has significant brain deteriorating features, including brain glucose hypometabolism, neuroinflammation, and leptin signaling disruption in the hippocampal area.

Light-emitting Diode Beam Profile and Spectral Output Influence on the Degree of Conversion of Bulk Fill Composites.

OBJECTIVES: To evaluate the beam profile and the spectral output of monowave and polywave light-emitting diodes (LEDs) and their influence on the degree of conversion (DC) of bulk fill composites. METHODS: A monowave LED (Smartlite Focus, Dentsply) and a polywave LED (Valo Cordless, Ultradent) were characterized using a resin calibrator and a laser beam profile analyzer. Two bulk fill composites, Sonic Fill 2 (SF) containing camphorquinone (CQ) and Tetric EvoCeram Bulk Fill (TEB) containing CQ associated with alternative photoinitiators, were placed in custom-designed molds (n=3) and photoactivated by the monowave or polywave LED with 20 J/cm2. To map the DC, longitudinal cross sections (0.5 mm thick) from the center of the restoration were evaluated using FT-NIR microscopy. SF and TEB light transmittances (n=3) through 4-mm-thick specimens were evaluated during curing. Data were analyzed using a split-plot analysis of variance and Tukey test (α=0.05; ß=0.2). RESULTS: The monowave LED had a radiant emittance of 20 ± 0.5 J/cm2 over 420-495 nm, and the polywave LED had an emittance of 15.5 ± 0.4 J/cm2 over 420-495 nm and of 4.5 ± 0.2 J/cm2 over 380-420 nm. The total radiant exposure at the bottom of TEB was 2.2 ± 0.2 J/cm2 with the monowave LED and 1.6 ± 0.3 J/cm2 with the polywave LED, and for SF it was 0.4 ± 0.1 J/cm2 for both LEDs. There were no differences in the curing profiles produced either by the monowave or the polywave LED (p=0.9), according to the regions under influence of blue and/or violet emission at the same depth. There was no statistical difference in the DC for SF using the monowave or polywave LED at any depth (p=0.29). TEB had a higher DC at up to 2 mm in depth when the polywave LED was used (p<0.004), but no differences were found when starting at 2.5 mm. CONCLUSIONS: Monowave and polywave LEDs emitted nonhomogeneous light beams, but this did not affect the DC homogeneity of bulk fill composites. For composites containing CQ associated with alternative photoinitiators, polywave LEDs had a higher DC, but only at the top part of the restoration; lower wavelength absorption photoinitiators were ineffective in deeper areas.

Isolation of sequences from a random-sequence expression library that mimic viral epitopes.

We describe the use of random peptide sequences for the mapping of antigenic determinants. An oligonucleotide with a completely degenerate sequence of 17 or 23 nucleotides was inserted into a bacterial expression vector. This resulted in an expression library producing random hexa- or octapeptides attached to a beta-galactosidase hybrid protein. Mimotopes, or antigenic sequences that mimic an epitope, were selected by immunoscreening of colonies with monoclonal antibodies, which were specific for antigenic sites on the spike protein of the coronavirus transmissible gastroenteritis virus. We report one mimotope for antigenic site II, eight for site III and one for site IV. The site III and site IV mimotopes were closely similar to the corresponding linear epitopes, localized previously in the amino acid sequence of the S protein. An alignment of the site II mimotope and the sequence of the S protein around Trp97, which is substituted in escape mutants, suggests that this mimotope mimics a conformational epitope located around residues 97-103. Applications of mimotopes to epitope mapping, serodiagnosis and vaccine development are discussed.

Antigenic structure of the E2 glycoprotein from transmissible gastroenteritis coronavirus.

The antigenic structure of transmissible gastroenteritis (TGE) virus E2 glycoprotein has been defined at three levels: antigenic sites, antigenic subsites and epitopes. Four antigenic sites (A, B, C and D) were defined by competitive radioimmunoassay (RIA) using monoclonal antibodies (MAbs) selected from 9 fusions. About 20% (197) of the hybridomas specific for TGE virus produced neutralizing MAbs specific for site A, which was one of the antigenically dominant determinants. Site A was differentiated in three antigenic subsites: a, b and c, by characterization of 11 MAb resistant (mar) mutants, that were defined by 8, 3, and 3 MAbs, respectively. These subsites were further subdivided in epitopes. A total of 11 epitopes were defined in E2 glycoprotein, eight of which were critical for virus neutralization. Neutralizing MAbs were obtained only when native virus was used to immunize mice, although to produce hybridomas mice immunizations were made with antigen in the native, denatured, or mixtures of native and denatured form. All neutralizing MAbs reacted to conformational epitopes. The antigenic structure of the E2-glycoprotein has been defined with murine MAbs, but the antigenic sites were relevant in the swine, the natural host of the virus, because porcine sera reacted against these sites. MAbs specific for TGE virus site C reacted to non-immune porcine sera. This reactivity was not directed against porcine immunoglobulins. These results indicated that TGE virus contains epitope(s) also present in some non-immunoglobulin component of porcine serum.

Lack of protection in vivo with neutralizing monoclonal antibodies to transmissible gastroenteritis virus.

Monoclonal antibodies (Mabs) specific for the E1 and E2 surface glycoproteins of the transmissible gastroenteritis virus (TGEV) of swine were examined either alone or in combination to evaluate their potential value in protecting neonatal pigs against a lethal dose of TGEV. Cesarean-delivered colostrum-deprived (CDCD) piglets were given one pre-challenge dose of Mab and an equal dose of the same Mab at each successive feeding after challenge. In vivo challenge results demonstrated that neither Mabs given individually nor combinations of the Mabs were able to protect neonatal pigs against a lethal dose of TGEV. However, in parallel experiments, polyclonal antibodies from immune colostrum or serum were protective.

Experiments on anthelmintic control of Fasciola hepatica in Brazil.

Two separate field trials involving naturally infected cattle were carried out on two farms known to have a history of Fasciola hepatica infection. On the first farm, 15 animals per group were allocated as follows: G1, triclabendazole (TCBZ) four times a year; G2, TCBZ twice a year (May and September); G3, untreated control. All groups grazed together and after 3.5 years the animals were slaughtered and their livers examined by federal meat inspectors who condemned 100% of livers in G3 and 8.3% in G2 owing to the presence of lesions of fasciolosis. In G1 no livers were condemned. Significant differences in weight gains were not detected and fluke counts remained at low levels in the treated groups. Also, in the control group, egg counts started to decrease when animals were 2 years old. On the second farm, groups of 20 animals were treated as follows: G1, TCBZ three times a year (May, September and December); G2, TCBZ twice a year (May and September); G3, nitroxynil twice a year (May and September); G4, rafoxanide twice a year (May and September); G5, untreated controls. All animals were weighed and faecal samples examined at approximately 28-day intervals. During the period of the study, larger weight gains were detected in the TCBZ treated groups than in the others. TCBZ treatment kept F. hepatica egg counts at a lower level for longer periods than the other drugs and significant differences in weight gains were only obtained between the group receiving TCBZ three times a year and the control group.

Degradation of propanil by bacterial isolates and mixed populations from a pristine lake.

The microbial transformation rates of propanil, a commonly used herbicide, were investigated using water from a pristine lake in northeast Georgia. Microbial degradation rates were measured using natural water microflora, the natural water microflora amended with five bacterial species (Aerobacter aerogenes, Aeromonas hydrophila, Acinetobacter calcoaceticus, Proteus mirabilis, and Aeromonas salmonicida) isolated from the same lake, and the five isolates individually. Transformation rate constants for propanil were compared for the mixed microbial assemblages and isolates at similar initial bacterial concentrations (approximately 5.0 x 10(-3) bacteria/mL). Degradation started within 60 hours and was completed by 160 hours in all experiments. The mean first-order rate constant for natural microflora was -(4.80 +/- 0.620) x 10(-3) h-1. Natural waters amended with the bacterial isolates yielded rate constants ranging from -(0.39 +/- 0.186) x 10(-3) h-1 to -(2.13 +/- 0.029) x 10(-3) h-1 with an overall mean of -(1.63 +/- 0.242) x 10(-3) h-1. After 660 hours following the first amendment of propanil, (i.e., 500 hours after propanil degradation was complete), each sample was again amended with propanil. Subsequent degradation rates ranged from -(21.3 +/- 0.186) x 10(-3) h-1 to -(64.2 +/- 0.786) x 10(-3) h-1 and the mean rate constant was -(37.5 +/- 0.922) x 10(-3) h-1. No significant differences were observed between first-order rate constants among isolates following the first or the second addition of propanil. After the second spike, however, the average of rate constants was approximately 20 times greater than that following the first spike. Rates for the individual isolates varied greatly from one isolate to another, ranging from virtually no degradation with A. calcoaceticus to -(21.6 +/- 0.332) x 10(-3) h-1 for the composite treatment of all isolates.

Age influence on quality of shell and penetration in bacterial laying lightweight eggs / Influência da idade na qualidade da casca e na penetração bacteriana em ovos de poedeiras leves

This study evaluated the influence of the bird's age on the quality of the shell and percentage of bacterial penetration in commercial eggs. White-shelled commercial eggs were used, laid by light laying hens in their first laying cycle at 21, 39, 51, and 62 weeks of age. Shell quality evaluations comprised: egg weight, specific gravity, percentage and thickness of shell, number and size of pores. For evaluations regarding bacterial penetration, strains of several enterobacterias and one salmonella were used, all of which resistant to Nalidixic acid (100µg/ml). The method employed for evaluation of bacterial penetration was filling the eggs with growth medium. The data were subjected to variance analysis with 5% of probability using SAS (Education Analytical Software, 2013). Eveb though increase in the laying hen's age caused reduction of the quality of eggshells, it failed to affect the percentage of penetration of the bacterial samples evaluated.(AU)

O presente estudo avaliou a influência da idade da ave sobre a qualidade da casca e na porcentagem de penetração bacteriana em ovos comerciais. Foram utilizados ovos brancos comerciais provenientes de poedeiras leves em primeiro ciclo de postura com 21, 39, 51 e 62 semanas de idade. As avaliações de qualidade da casca realizadas foram: peso do ovo, gravidade específica, porcentagem e espessura da casca, número e tamanho dos poros. Para as avaliações da penetração bacteriana, foram utilizadas cepas de diversas enterobactérias e uma salmonela, sendo todas resistentes ao ácido nalidíxico (100µg/mL). O método utilizado para a avaliação da penetração bacteriana foi por meio do preenchimento dos ovos com meio de cultura. Os dados foram submetidos à análise de variância com 5% de probabilidade utilizando-se o programa SAS - Statistical Analisys System (Education Analytical Software, 2013). O aumento da idade da poedeira promoveu a redução da qualidade da casca dos ovos, porém não foi capaz de influenciar a porcentagem de penetração das amostras bacterianas avaliadas.(AU)

Update on HER-2 as a target for cancer therapy: HER2/neu peptides as tumour vaccines for T cell recognition.

During the past decade there has been renewed interest in the use of vaccine immunotherapy for the treatment of cancer. This review focuses on HER2/neu, a tumour-associated antigen that is overexpressed in 10-40% of breast cancers and other carcinomata. Several immunogenic HER2/neu peptides recognized by T lymphocytes have been identified to be included in cancer vaccines. Some of these peptides have been assessed in clinical trials of patients with breast and ovarian cancer. Although it has been possible to detect immunological responses against the peptides in the immunized patients, no clinical responses have so far been described. Immunological tolerance to self-antigens like HER2/neu may limit the functional immune responses against them. It will be of interest to determine whether immune responses against HER2/neu epitopes can be of relevance to cancer treatment.

Binding of diverse peptides to MHC class I molecules inhibits target cell lysis by activated natural killer cells.

Class I MHC expression by target cells inhibits lysis mediated by natural killer (NK) cells, often in an allele-specific fashion. It has been proposed that NK cell inhibitory receptors recognize complexes of class I molecules with specific cellular peptides that define self, displacement of which would render cells NK sensitive. By loading the mostly empty Dd class I molecules of cell lines deficient in peptide transporter molecules with synthetic or natural Dd-bound peptides, we have demonstrated specific dose-dependent inhibition of the Ly49+ subset of activated NK cells by class I-peptide complexes. Inhibition occurred with most if not all Dd-binding peptides, suggesting that Ly49+ NK cells recognize class I-peptide complexes largely independently of peptide composition. The results suggest a primary role of NK cells in the destruction of cells that have down-regulated or extinguished cell surface expression of some or all class I molecules.

Dynamic holographic moiré patterns in photorefractive Bi12TiO20 and small-angle measurements.

A procedure for obtaining real-time holographic moirélike patterns and measuring small angles is proposed. Two rotated sinusoidal phase gratings are superposed, and the result represents a promising technique for making small-angle measurements in metrological applications. The experiments are performed with a diffusion-only recording mechanism in the photorefractive crystal Bi12TiO20 illuminated by lambda = 0.633 microm light from a He-Ne laser.

Multiple natural killer cell-activating signals are inhibited by major histocompatibility complex class I expression in target cells.

Several lines of evidence indicate that major histocompatibility complex class I molecules expressed by target cells can prevent natural killer cell (NK) lysis, possibly by engaging inhibitory receptors expressed by NK cells. On the other hand it is likely that NK cells must be activated to lysis by the recognition of unidentified NK target structures on target cells. To investigate the relationship between positive activation of NK cells by NK target structures versus inhibition by target cell class I molecules, we have examined various NK/target cell interactions for which the expression of inhibitory class I molecules by the target cells is known. The results suggests that specific properties of the target cell other than the absence of class I expression are necessary to activate NK-mediated lysis. Furthermore, different effector cell populations, i.e. freshly isolated versus interleukin-2 activated NK cells, differ in their capacity to kill class I-deficient lymphoblast target cells. In general, class I-deficient target cells that are resistant to direct lysis by a given NK population can be lysed by the NK cells when the reaction is mediated by antibody-dependent cellular cytotoxicity (ADCC). Most significantly, all types of NK-mediated lysis of lymphoblasts, of tumor cells and of almost any target by ADCC can be inhibited by appropriate class I gene expression in the target cell. These results suggest a model in which lysis by NK cells must be triggered by any one of a set of distinct target cell ligands, but that all of these signals can be overruled by class I-mediated inhibition.

Antigenic differentiation between transmissible gastroenteritis virus of swine and a related porcine respiratory coronavirus.

The antigenic relationship between a recently isolated porcine respiratory coronavirus (TLM 83) and transmissible gastroenteritis (TGE) virus of swine was studied by neutralization, immunoblotting and radioimmunoassay (RIA) using TGE virus-specific monoclonal antibodies (MAbs) and polyclonal antibodies specific for both viruses. A complete two-way neutralization activity between the two viruses was found. Immunoblotting revealed cross-reactions between TLM 83 and TGE virus antigens at the level of the envelope protein (E1), the nucleoprotein (N) and the peplomer protein (E2). By virus neutralization assays and RIA with TGE virus-specific MAbs, the presence of similar epitopes in the E1 and N proteins and in the neutralization-mediating antigenic site of the E2 protein were demonstrated. E2 protein-specific MAbs, without neutralizing activity and reacting with antigenic sites B, C and D (previously defined), failed to recognize TLM 83. These results indicated a close antigenic relationship and structural similarity between TLM 83 and TGE viruses and also suggested potential ways of differentiating between the two viruses.