Identification of assembly precursors to photosystems emitting fluorescence at 683 nm and 687 nm by cryogenic fluorescence microspectroscopy

Photosystem I (PSI) and photosystem II (PSII) play key roles in photoinduced electron-transfer reaction in oxygenic photosynthesis. Assemblies of these PSs can be initiated by illumination of the etiolated seedlings (greening). The study aimed to identify specific fluorescence spectral components relevant to PSI and PSII assembly intermediates emerging in greening seedlings of Zea mays, a typical C4 plant. The different PSII contents between the bundle sheath (BS) and mesophyll (M) cells were utilized to spectrally isolate the precursors to PSI and PSII. The greening Zea mays leaf thin sections were observed with the cryogenic microscope combined with a spectrometer. With the aid of the singular-value decomposition analysis, we could identify four independent fluorescent species, SAS677, SAS685, SAS683, and SAS687, named after their fluorescence peak wavelengths. SAS677 and SAS685 are the dominant components after the 30-minute greening, and the distributions of these components showed no clear differences between M and BS cells, indicating immature cell differentiation in this developing stage. On the other hand, the 1-hour greening resulted in reduced distributions of SAS683 in BS cells leading us to assign this species to PSII precursors. The 2-hour greening induced the enrichment of SAS687 in BS cells suggesting its PSI relevance. Similarity in the peak wavelengths of SAS683 and the reported reaction center of PSII implied their connection. SAS687 showed an intense sub-band at around 740 nm, which can be assigned to the emission from the red chlorophylls specific to the mature PSI.     

β-Glucuronidase activity in human T and B lymphocytes and the Tμ and Tγ subpopulations

Summary The -glucuronidase staining characteristics of isolated T cell populations and the T and T enriched fractions derived of them were studied. T lymphocytes were obtained from purified T lymphocytes by ox-IgG rosette sedimentation. The rosette-forming cells in the pellet were referred to as T lymphocytes, whereas the lymphocytes in the interface were referred to as T depleted or T lymphocytes. B cells were studied on rosetted mononuclear cells with either mouse erythrocytes or with Staphylococcus Aureus (Cowan I) bacteria, after a preceeding polyvalent anti-human Ig treatment of the cells. While B cells showed mostly no reactivity, T and T cells were respectively characterised by a dot-like and granular pattern of reactivity. These findings are in agreement with those observed by others after -naphthyl-acetate esterase or acid phosphatase staining. Within the T lymphocyte fraction, the T non-, non lymphocytes seemed to have a granular pattern of reactivity. The same staining pattern was found in non-B, non-T lymphocytes.     

Monte carlo model and output factors of elekta infinity™ 6 and 10 MV photon beam

AimThis study aimed to commission the Elekta Infinity™ working in 6 and 10 MV photon beam installed in Concord International Hospital, Singapore, and compare the OFs between MC simulation and measurement using PTW semiflex and microDiamond detector for small field sizes.Material and MethodsThere are two main steps in this study: modelling of Linac 6 and 10 MV photon beam and analysis of the output factors for field size 2 × 2–10 × 10 cm². The EGSnrc/BEAMnrc-DOSXYZnrc code was used to model and characterize the Linac and to calculate the dose distributions in a water phantom. The dose distribution and OFs were compared to the measurement data in the same condition.ResultsThe commissioning process was only conducted for a 10 × 10 cm² field size. The PDD obtained from MC simulation showed a good agreement with the measurement. The local dose difference of PDDs was less than 2% for 6 and 10 MV. The initial electron energy was 5.2 and 9.4 MeV for 6 and 10 MV photon beam, respectively. This Linac model can be used for dose calculation in other situations and different field sizes because this Linac has been commissioned and validated using Monte Carlo simulation. The 10 MV Linac produces higher electron contamination than that of 6 MV.ConclusionsThe Linac model in this study was acceptable. The most important result in this work comes from OFs resulted from MC calculation. This value was more significant than the OFs from measurement using semiflex and microDiamond for all beam energy and field sizes because of the CPE phenomenon.     

Development of novel liver X receptor modulators based on a 1,2,4-triazole scaffold

Liver?X?Receptor (LXR) agonists have been reported as a potential treatment for atherosclerosis, Alzheimer’s disease and hepatitis C virus (HCV) infection. We have designed and synthesized a series of potent compounds based on a 1,2,4-triazole scaffold as novel LXR modulators. In cell-based cotransfection assays these compounds generally functioned as LXR agonists and we observed compounds with selectivity towards LXRα (7-fold) and LXRβ (7-fold) in terms of potency. Assessment of the effects of selected compounds on LXR target gene expression in HepG2 cells revealed that compounds 6a-b and 8a-b behaved as inverse agonists on FASN expression even though they were agonists in the LXRα and LXRβ cotransfection assays. Interestingly, these compounds had no effect on the expression of SREBP-1c confirming a unique LXR modulator pharmacology. Molecular docking studies and evaluation of ADME properties in-silico show that active compounds possess favorable binding modes and ADME profiles. Thus, these compounds may be useful for in vivo characterization of LXR modulators with unique profiles and determination of their potential clinical utility.     

Supplemental L-arginine improves feed conversion and modulates lipid metabolism in male and female broilers from 29 to 42 days of age

The excessive accumulation of abdominal fat in broilers is an economic waste. Supplemental dietary L-arginine has been shown to reduce lipogenesis in broilers, but studies on this subject are still scarce. Two experiments were conducted in a 5 × 2 factorial design, with five L-arginine concentrations in diets (0, 3, 6, 9 and 12 g/kg) provided to male and female broilers, from 29 to 42 days of age, reared in boxes or cages, with six replicates of 23 broilers per box and six replicates of three broilers per cage, totalling 1560 broilers. Data on performance, carcass and cuts yield, abdominal fat deposition, chemical composition of the breast, lipid profile and liver enzyme activity were evaluated in experiment 1. In experiment 2, the balance and retention of nitrogen, metabolizability coefficients and metabolizable energy of feed were determined and the treatments were evaluated in six replicates of three broilers per cage, totalling 180 broilers. In both experiments, there was no interaction (P > 0.05) between L-arginine concentration in the diet and sex of the broilers for all parameters evaluated. Male broilers showed higher (P < 0.05) weight gain, noble cuts yield, levels of CP and mineral matter in the breast, better feed conversion (P < 0.05) and lower (P < 0.05) malic enzyme activity in the liver and abdominal fat deposition. On the other hand, regardless of sex, the increase in L-arginine concentration in the diet improved (P < 0.05) the feed conversion in addition to reducing (P < 0.05) serum levels of total cholesterol and low-density lipoprotein cholesterol, the malic enzyme activity in the liver and abdominal fat deposition. In conclusion, male broilers had better productive results than females. However, supplementing the diet of male or female broilers from 29 to 42 days of age with L-arginine at a concentration of 6.87 g/kg represents a nutritional strategy to improve feed conversion and reduce circulating triacylglycerol and cholesterol levels, NADPH synthesis by liver malic enzyme and abdominal fat deposition, without negatively affecting the carcass and noble cuts yield, the amount of nitrogen excreted by the broilers and the energy value of the feed.     

Fast scanning calorimetry of lysozyme unfolding at scanning rates from 5 K/min to 500,000 K/min

Background

Protein denaturation is often studied using differential scanning calorimetry (DSC). However, conventional instruments are limited in the temperature scanning rate available. Fast scanning calorimetry (FSC) provides an ability to study processes at much higher rates while using extremely small sample masses [ng]. This makes it a very interesting technique for protein investigation.

Persistent Changes in Circulating and Intestinal γδ T Cell Subsets,Invariant Natural Killer T Cells and Mucosal-Associated Invariant T Cells in Children and Adults with Coeliac Disease

Coeliac disease is a chronic small intestinal immune-mediated enteropathy precipitated by exposure to dietary gluten in genetically predisposed individuals. The only current therapy is a lifelong gluten free diet. While much work has focused on the gliadin-specific adaptive immune response in coeliac disease, little is understood about the involvement of the innate immune system. Here we used multi-colour flow cytometry to determine the number and frequency of γδ T cells (Vδ1, Vδ2 and Vδ3 subsets), natural killer cells, CD56⁺ T cells, invariant NKT cells, and mucosal associated invariant T cells, in blood and duodenum from adults and children with coeliac disease and healthy matched controls. All circulating innate lymphocyte populations were significantly decreased in adult, but not paediatric coeliac donors, when compared with healthy controls. Within the normal small intestine, we noted that Vδ3 cells were the most abundant γδ T cell type in the adult epithelium and lamina propria, and in the paediatric lamina propria. In contrast, patients with coeliac disease showed skewing toward a predominant Vδ1 profile, observed for both adult and paediatric coeliac disease cohorts, particularly within the gut epithelium. This was concurrent with decreases in all other gut lymphocyte subsets, suggesting a specific involvement of Vδ1 cells in coeliac disease pathogenesis. Further analysis showed that γδ T cells isolated from the coeliac gut display an activated, effector memory phenotype, and retain the ability to rapidly respond to in vitro stimulation. A profound loss of CD56 expression in all lymphocyte populations was noted in the coeliac gut. These findings demonstrate a sustained aberrant innate lymphocyte profile in coeliac disease patients of all ages, persisting even after elimination of gluten from the diet. This may lead to impaired immunity, and could potentially account for the increased incidence of autoimmune co-morbidity.     

Permanent seed implant brachytherapy in low-risk prostate cancer: Preoperative planning with 145 Gy versus real-time intraoperative planning with 160 Gy

Aim

The present retrospective study was to compare toxicity and survival outcomes in a group of low-risk PCa patients treated with either the preoperative planning technique (145?Gy) or the real-time IoP technique (160?Gy).

Hypoxia-increased RAGE expression regulates chemotaxis and pro-inflammatory cytokines release through nuclear translocation of NF-κ B and HIF1α in THP-1 cells

The potential role of hypoxia in mediating the receptor for advanced glycation end products (RAGE) expression deserves to be confirmed. And the role of RAGE in hypoxia-induced chemotaxis and inflammation is still unclear. In present study, THP-1?cells were pretreated with siRNA to block HIF1α, NF-κ B, or RAGE, followed by exposed to hypoxia (combined with H₂O₂ or SNP), and then RAGE expression, nuclear translocation of HIF1α and NF-κ B, release of TNF-α and IL-1β, as well as expression of MCP-1 and CCR2 were measured. The results revealed that RAGE mRNA and protein in THP-1?cells were significantly increased after exposed into hypoxia atmosphere, especially into the solution containing SNP or H₂O₂. Moreover, SNP or H₂O₂ exposure could further amplify hypoxia-induced nuclear translocation of HIF-1α and NF-κ B. Knockdown HIF-1α or NF-κ B by siRNAs could reduce hypoxia- and oxidative stress-induced RAGE hyper-expression. And pretreatment THP-1?cells with RAGE siRNA or NF-κ B siRNA could reduce hypoxia- and oxidative stress-induced expression of MCP-1 and CCR2, and release of TNF-α and IL-1β. Thus, hypoxia not only increases RAGE expression in THP-1?cells by promoting nuclear translocation of NF-κ B and HIF1α, but also regulates chemotaxis and pro-inflammatory cytokines release, which may be partially mediated through upregulation of RAGE expression.     

A virus-specified mechanism for the prevention of multiple infection—T7- and T3-mutual and superinfection exclusion

Summary Co-and superinfection of cells with T3/T7 result in exclusion (mutual or superinfection exclusion). The exclusion mechanism is also directed against homologous (or identical) virus. Exclusion is established after the adsorption but before the genome becomes available for gene expression or replication, that is only one virus per cell develops. The exclusion is triggered by a constituant of the viral particle. An early T7 gene (M gene) (Schweiger et al., 1975) is essential for the formation of exclusion competent virions.     

A genome-wide perspective on the diversity and selection signatures in indigenous goats using 53 K single nucleotide polymorphism array

Tibetan goats, Taihang goats, Jining grey goats, and Meigu goats are the representative indigenous goats in China, found in Qinghai-Tibet Plateau, Western pastoral area, Northern and Southern agricultural regions. Very few studies have conducted a comprehensive analysis of the genomic diversity and selection of these breeds. We genotyped 96 unrelated individuals, using goat 53 K Illumina BeadChip array, of the following goat breeds: Tibetan (TG), Taihang (THG), Jining grey (JGG), and Meigu (MGG). A total of 45 951 single nucleotide polymorphisms were filtered to estimate the genetic diversity and selection signatures. All breeds had a high proportion (over 95%) of polymorphic loci. The observed and excepted heterozygosity ranged from 0.338 (MGG) to 0.402 (JGG) and 0.339 (MGG) to 0.395 (JGG), respectively. Clustering analysis displayed a genetically distinct lineage for each breed, and their Fst were greater than 0.25, indicating that they had a higher genetic differentiation between groups. Furthermore, effective population size reduced in all four populations, indicating a loss of genetic diversity. In addition, runs of homozygosity were mainly distributed in 5–10 Mb. Lastly, we identified signature genes, which were closely related to high-altitude adaptation (ADIRF) and prolificity (CNTROB, SMC3, and PTEN). This study provides a valuable resource for future studies on genome-wide perspectives on the diversity and selection signatures of Chinese indigenous goats.     

Occurrence and characterization of Acanthamoeba similar to genotypes T4, T5, and T2/T6 isolated from environmental sources in Brasília, Federal District, Brazil

Species of Acanthamoeba can cause keratitis and brain infections. The characterization of environmental isolates is necessary to analyze the risk of human infection. We aimed at identifying and genotyping Acanthamoeba isolates from soil, swimming pools, and water features in Brasília, Federal District, Brazil, as well as determining their physiological characteristics and pathogenic potential. Among the 18 isolates studied, eight were similar to genotype T5, five to T4, and one to T2/T6, classified by the sequence analysis of 18S rDNA. Genotypes of four isolates were not determined. Ten isolates (55%) grew at 37 °C and seven (39%) grew in media with 1.5M mannitol, which are the physiological parameters associated with pathogenic Acanthamoeba; also, four isolates from swimming pools presented high pathogenic potential. Our results indicate a widespread distribution of potentially pathogenic Acanthamoeba T4, T5, and T2/T6 in different environmental sources in Brasília, revealing the potential risk of human infection and the need of preventive measures.     

VEGF₁₆₄ differentially regulates neutrophil and T cell adhesion through ItgaL- and ItgaM-dependent mechanisms

Leukocyte recruitment to inflamed tissues is the cornerstone of inflammatory responses and the driving force behind the establishment of inflammatory bowel disease, consisting of Crohn's disease and ulcerative colitis. It has been reported that angiogenic cytokines contribute to this inflammatory response that facilitates the chronic nature of disease. We have previously reported (Goebel S, Huang M, Davis WC, Jennings M, Siahaan TJ, Alexander JS, Kevil CG. Am J Physiol Gastrointest Liver Physiol 290: G648-G654, 2006) that vascular endothelial growth factor (VEGF)-A can stimulate neutrophil adhesion to colon microvascular endothelial cells in a β?-integrin (Itgb2)-dependent manner. However, it is not known which of the specific leukocyte integrins are critical for VEGF-A-dependent neutrophil and T cell recruitment. Here we examine the differential importance of either α-integrin (Itga)L or ItgaM in governing neutrophil and T cell adhesion to VEGF-A-activated colonic endothelium. Using an in vitro parallel-plate flow chamber model, we found that genetic deficiency of ItgaM completely blunted neutrophil adhesion to VEGF-A-stimulated endothelium, whereas ItgaL deficiency only partly blocked neutrophil adhesion. Deficiency of ItgaM did significantly decrease neutrophil rolling, whereas deficiency of ItgaL did not. We found that genetic deficiency of either ItgaL or ItgaM did significantly blunt T cell adhesion to VEGF-A-stimulated colon endothelium. We also found that genetic deficiency of these Itgas significantly attenuated T cell rolling behavior. Lastly, we examined whether VEGF-A-mediated leukocyte recruitment occurred through different VEGF receptor (VEGFR) pathways and found that VEGFR2 activation regulates neutrophil recruitment, whereas both VEGFR1 and VEGFR2 modulate T cell recruitment. Together, these data identify differential molecular mechanisms of VEGF-A-mediated leukocyte recruitment.     

Extended transition milk feeding for 3 weeks improves growth performance and reduces the susceptibility to diarrhea in newborn female Holstein calves

Dairy calves may benefit from extending the duration of feeding transition milk (TM; the subsequent two to six milkings after parturition) to enhance performance and health during early life. The objective of this study was to assess the effect of replacing pasteurized waste milk (non-saleable milk containing antibiotic and/or drug residues) with pasteurized TM for 3 weeks on the growth performance and health of dairy calves. A total of 84 healthy newborn female Holstein calves were blocked by birth order and assigned randomly to 4 treatment groups with partial replacement of pasteurized waste milk by TM (second milkings after parturition) at 0 (0 l/day TM + 6 l/day milk), 0.5 (0.5 l/day TM + 5.5 l/day milk), 1 (1 l/day TM + 5 l/day milk), or 2 l (2 l/day TM + 4 l/day milk) for a 21-day period. From day 22 onward, all calves were fed individually with 6 l/day pasteurized waste milk. Calves were weaned on day 60 and monitored until day 90 of the study. Liquid feed DM intake (DMI) was increased with increasing levels of TM (P = 0.001). Starter feed DMI and total DMI (liquid feed DMI + starter feed DMI) were not affected by the treatment effect. Calves were fed 2 l/day TM gained more BW compared with those in the control group during the postweaning and overall periods. The average daily gain tended (P = 0.06) to be higher in calves fed 2 l/day TM compared with calves fed 0 (+ 65 g/day), 0.5 (+ 53 g/day), or 1 (+ 76 g/day) l/day TM during the preweaning period. Daily weight gain was also higher in calves fed 2 l/day TM compared with calves in the control group during the postweaning (+ 137 g/day; P = 0.04) and overall (+ 89 g/day; P < 0.01) periods, respectively. Calves fed 2 l/day TM had a higher feed efficiency compared with calves in the control group during all studied periods. The calves fed TM2 had a lower chance of having diarrhea compared with other treatment groups. Duration but not the frequency of diarrhea was lower in calves fed TM2 vs TM0 (2.5 vs 4.2 days; P = 0.03). In general, partial replacement of waste milk with TM (2 l/day) may be recommended to feed dairy calves at an early stage of life to support a higher growth rate and health benefits.     

Fluorine-18 (18F)-labeled retinoid x receptor (RXR) partial agonist whose tissue transferability is affected by other RXR ligands

Bexarotene (1), a retinoid X receptor (RXR) agonist approved for the treatment of cutaneous T cell lymphoma (CTCL), was reported to migrate into baboon brain based on findings obtained by positron emission tomography (PET) with a ¹¹C-labeled tracer. However, co-administration of non-radioactive 1 had no effect on the distribution of [¹¹C]1, probably due to non-specific binding of 1 as a result of its high lipophilicity. Here, we report a fluorine-18 (¹⁸F)-labeled PET tracer [¹⁸F]6 derived from RXR partial agonist CBt-PMN (2), which has lower lipophilicity and weaker RXR-binding ability than [¹¹C]1. The concomitant administration of 1 or 2 with [¹⁸F]6 with resulted in decreased accumulation of [¹⁸F]6 in liver, together with increased brain uptake and increased accumulation in kidney and muscle, as visualized by PET. A plausible explanation of these findings is the inhibition of [¹⁸F]6 uptake into the liver by concomitantly administered 1 or 2, leading to an increase in blood concentration of [¹⁸F]6 followed by increased accumulation in other tissues.