FoxP3+ regulatory T cells essentially contribute to peripheral CD8+ T-cell tolerance induced by steady-state dendritic cells

Peripheral T-cell tolerance is thought to significantly contribute to the prevention of autoimmunity, and it has been shown that antigen-presenting steady-state dendritic cells efficiently induce peripheral tolerance. We previously showed that dendritic-cell–induced tolerance is a T-cell–intrinsic process that depends on coinhibitory molecules such as programmed death-1. Here we specifically analyze the involvement of FoxP3⁺ regulatory T cells, which are known to be important for maintenance of self-tolerance. We show that antigen presentation by steady-state dendritic cells failed to induce peripheral tolerance in the absence of FoxP3⁺ regulatory T cells but induced protective CD8⁺ T-cell–mediated immunity instead. Regulatory T-cell–depleted mice had massively increased numbers of dendritic cells in lymph nodes. Dendritic cells isolated from mice without regulatory T cells had up-regulated costimulatory molecules and showed stronger T-cell stimulatory capacity ex vivo, suggesting that regulatory T cells contribute to peripheral tolerance by keeping the dendritic cells in an immature state. Using blocking antibodies, we demonstrate that CTLA-4 but not IL-10 is necessary for control of dendritic cells by regulatory T cells.     

X-Nuclei NMR Self-Diffusion Studies in Mesoporous Silica Foam and Microporous MOF CuBTC

A standard X-observe NMR probe was equipped with a z-gradient coil to enable high-sensitivity pulsed field gradient NMR diffusion studies of Li⁺ and Cs⁺ cations of aqueous salt solutions in a high-porosity mesocellular silica foam (MCF) and of CO₂ adsorbed in metal-organic frameworks (MOF). The coil design and the necessary probe modifications, which yield pulsed field gradients of up to ±16.2 Tm^-1, are introduced. The system was calibrated at ²H resonance frequency and successfully applied for diffusion studies at ⁷Li, ²³Na, ¹³C and ¹³³Cs frequencies. Significant reductions of the diffusivities of the cations in LiCl_ac and CsCl_ac solution introduced into MCFs are observed. By comparison of the diffusion behavior with the bulk solutions, a tortuosity of the silica foam of 4.5 ± 0.6 was derived. Single component self-diffusion of CO₂ and CH₄ (measured by ¹H NMR) as well as self-diffusion of the individual components in CO₂/CH₄ mixtures was studied in the MOF CuBTC. The experimental results confirm high mobilities of the adsorbed gases and trends for diffusion separation factors predicted by MD simulations.     

Placement of 125I seed strands and stents for a type IV Klatskin tumor

Herein, we report a new technique that consists of placing two ¹²⁵I seed strands and two stents in the right and left intrahepatic bile ducts for the treatment of hilar cholangiocarcinoma. A 75-year-old man presented with jaundice and was diagnosed with Bismuth type IV Klatskin tumor. Abdominal computed tomography (CT) showed intrahepatic and extrahepatic bile duct dilatation and a soft tissue mass in the hepatic hilum. Because curative surgical resection was not possible, we placed ¹²⁵I seed strands and stents in the right and left intrahepatic bile ducts. Three months later, abdominal CT showed less intrahepatic and extrahepatic bile duct dilatation than before the procedure. This technique was feasible and could be considered for the treatment of patients with Bismuth type IV tumors.     

Cellulose Isolation Methodology for NMR Analysis of Cellulose Ultrastructure

In order to obtain accurate information about the ultrastructure of cellulose from native biomass by ¹³C cross polarization magic angle spinning (CP/MAS) NMR spectroscopy the cellulose component must be isolated due to overlapping resonances from both lignin and hemicellulose. Typically, cellulose isolation has been achieved via holocellulose pulping to remove lignin followed by an acid hydrolysis procedure to remove the hemicellulose components. Using ¹³C CP/MAS NMR and non-linear line-fitting of the cellulose C₄ region, it was observed that the standard acid hydrolysis procedure caused an apparent increase in crystallinity of ~10% or less on the cellulose isolated from Populus holocellulose. We have examined the effect of the cellulose isolation method, particularly the acid treatment time for hemicellulose removal, on cellulose ultrastructural characteristics by studying these effects on cotton, microcrystalline cellulose (MCC) and holocellulose pulped Populus. ¹³C CP/MAS NMR of MCC indicated that holocellulose pulping and acid hydrolysis has little effect on the crystalline ultrastructural components of cellulose. Although any chemical method to isolate cellulose from native biomass will invariably alter substrate characteristics, especially those related to regions accessible to solvents, we found those changes to be minimal and consistent in samples of typical crystallinity and lignin/hemicellulose content. Based on the rate of the hemicellulose removal, as determined by HPLC-carbohydrate analysis and magnitude of cellulose ultrastructural alteration, the most suitable cellulose isolation methodology utilizes a treatment of 2.5 M HCl at 100 °C for a standard residence time between 1.5 and 4 h. However, for the most accurate crystallinity results this residence time should be determined empirically for a particular sample.     

3.0T 1H magnetic resonance spectroscopy for assessment of steatosis in patients with chronic hepatitis C

AIM: To investigate the utility of ¹H magnetic resonance spectroscopy (¹H MRS) as a noninvasive test for steatosis in patients infected with hepatitis C virus.METHODS: Ninety patients with chronic hepatitis C and pathology data underwent 3.0T ¹H MRS, and the results of MRS and pathological analysis were compared.RESULTS: This group of patients included 26 people with mild fatty liver (28.89%), 16 people with moderate fatty liver (17.78%), 18 people with severe fatty liver (20.0%), and 30 people without fatty liver (33.33%). The water peak was near 4.7 parts per million (ppm), and the lipid peak was near 1.3 ppm. Analysis of variance revealed that differences in the lipid peak, the area under the lipid peak, ratio of the lipid peak to the water peak, and ratio of the area under the lipid peak to the area under the water peak were statistically significant among the groups. Specifically, as the severity of fatty liver increased, the value of each index increased correspondingly. In the pairwise comparisons, the mean lipid peak, area under the lipid peak, ratio of the lipid peak to the water peak, and ratio of the area under the lipid peak to the area under the water peak were significantly different between the no fatty liver and moderate fatty liver groups, whereas no differences were noted between the severe fatty liver group and the mild or moderate fatty liver group. Area under the ROC curve (AUC) of area ratio in lipid and water and ratio in lipid and water in the no fatty liver group to mild fatty liver group, mild fatty liver group to moderate fatty liver group, and moderate fatty liver disease group to severe fatty liver group, were 0.705, 0.900, and 0.975, respectively.CONCLUSION: ¹H MRS is a noninvasive technique that can be used to provide information on the effect of liver steatosis on hepatic metabolic processes. This study indicates that the ¹H MRS can be used as an indicator of steatosis in patients with chronic hepatitis C.     

Quantifying Susceptibility of CD4+ Stem Memory T-Cells to Infection by Laboratory Adapted and Clinical HIV-1 Strains

CD4⁺ T cells are principal targets for human immunodeficiency virus type 1 (HIV-1) infection. CD4⁺ T cell subsets are heterogeneous cell populations, divided by functional and phenotypic differences into naïve and memory T cells. The memory CD4⁺ T cells are further segregated into central, effector and transitional memory cell subsets by functional, phenotypic and homeostatic characteristics. Defining the distribution of HIV-1 infection in different T cell subsets is important, as this can play a role in determining the size and composition of the viral reservoir. Both central memory and transitional memory CD4⁺ T cells have been described as long-lived viral reservoirs for HIV. Recently, the newly described stem memory T cell subset has also been implicated as a long-lived HIV reservoir. Using green fluorescent protein (GFP) reporter strains of HIV-1 and multi parameter flow cytometry, we developed an assay to simultaneously quantify the susceptibility of stem memory (TSCM), central memory, effector memory, transitional memory and naïve CD4⁺ T cell subsets, to HIV-1 infection in vitro. We show that TSCM are susceptible to infection with laboratory adapted and clinical HIV-1 strains. Our system facilitates the quantitation of HIV-1 infection in alternative T cell subsets by CCR5- and CXCR4-using viruses across different HIV-1 subtypes, and will be useful for studies of HIV-1 pathogenesis and viral reservoirs.     

Control of seed mass and seed yield by the floral homeotic gene APETALA2

APETALA2 (AP2) is best known for its role in the regulation of flower meristem and flower organ identity and development in Arabidopsis. We show here that AP2 also plays an important role in determining seed size, seed weight, and the accumulation of seed oil and protein. We demonstrate genetically that AP2 acts through the maternal sporophyte and endosperm genomes to control seed weight and seed yield. Thus, AP2 functions outside the boundaries of flower meristem and flower organ development to affect agronomically relevant traits in Arabidopsis.     

Characteristics of Classical Swine Fever Virus Variants Derived from Live Attenuated GPE− Vaccine Seed

The GPE⁻ strain is a live attenuated vaccine for classical swine fever (CSF) developed in Japan. In the context of increasing attention for the differentiating infected from vaccinated animals (DIVA) concept, the achievement of CSF eradication with the GPE⁻ proposes it as a preferable backbone for a recombinant CSF marker vaccine. While its infectious cDNA clone, vGPE⁻, is well characterized, 10 amino acid substitutions were recognized in the genome, compared to the original GPE⁻ vaccine seed. To clarify the GPE⁻ seed availability, this study aimed to generate and characterize a clone possessing the identical amino acid sequence to the GPE⁻ seed. The attempt resulted in the loss of the infectious GPE⁻ seed clone production due to the impaired replication by an amino acid substitution in the viral polymerase NS5B. Accordingly, replication-competent GPE⁻ seed variant clones were produced. Although they were mostly restricted to propagate in the tonsils of pigs, similarly to vGPE⁻, their type I interferon-inducing capacity was significantly lower than that of vGPE⁻. Taken together, vGPE⁻ mainly retains ideal properties for the CSF vaccine, compared with the seed variants, and is probably useful in the development of a CSF marker vaccine.     

1O2-mediated retrograde signaling during late embryogenesis predetermines plastid differentiation in seedlings by recruiting abscisic acid

Plastid development in seedlings of Arabidopsis thaliana is affected by the transfer of ¹O₂-mediated retrograde signals from the plastid to the nucleus and changes in nuclear gene expression during late embryogenesis. The potential impact of these mechanisms on plastid differentiation is maintained throughout seed dormancy and becomes effective only after seed germination. Inactivation of the 2 nuclear-encoded plastid proteins EXECUTER1 and EXECUTER2 blocks ¹O₂-mediated retrograde signaling before the onset of dormancy and impairs normal plastid formation in germinating seeds. This long-term effect of ¹O₂ retrograde signaling depends on the recruitment of abscisic acid (ABA) during seedling development. Unexpectedly, ABA acts as a positive regulator of plastid formation in etiolated and light-grown seedlings.     

The tolerance–fecundity trade-off and the maintenance of diversity in seed size

Seed size commonly varies by five to six orders of magnitude among coexisting plant species, a pattern ecologists have long sought to explain. Because seed size trades off with seed number, small-seeded species clearly have the advantage in fecundity, but what is the countervailing advantage of large seeds? Higher competitive ability combined with strong competitive asymmetry can in theory allow coexistence through a competition–colonization trade-off, but empirical evidence is inconsistent with this mechanism. Instead, the key advantage of large seeds appears to be their tolerance of stresses such as shade or drought that are present in some but not all regeneration sites. Here I present a simple, analytically tractable model of species coexistence in heterogeneous habitats through a tolerance–fecundity trade-off. Under this mechanism, the more tolerant species win all of the more stressful regeneration sites and some of those that are less stressful, whereas the more fecund species win most but not all of the less stressful sites. The tolerance–fecundity trade-off enables stable coexistence of large numbers of species in models with and without seed limitation. The tolerance–fecundity mechanism provides an excellent explanation for the maintenance of diversity of seed size within plant communities and also suggests new hypotheses for coexistence in animal and microbial communities.     

To evaluate the efficacy and safety of 125I seed implantation in SCLC as second line therapy

This study aimed to evaluate the efficacy and safety of iodine 125 (¹²⁵I) radioactive seed implantation for small cell lung cancer at the limited stage of relapse as second line therapy. We collected 6 patients with recurred limited stage small cell lung cancer, who got pathological diagnosis after a bronchoscopic biopsy and then received standard first line treatment, combined chemotherapy and radiotherapy, including prophylactic cranial irradiation. These recurred small cell lung cancer patients got ¹²⁵I seed implantation treatment as second line therapy, if the treatment not good responsive or the disease got rapid progress, we used the second line chemotherapy as salvage treatment. Clinical data of these patients were collected and short-term effects were observed. The follow-up period lasted for 42 months. All the patients tolerated the procedure of ¹²⁵I radioactive seed implantation very well. We followed up the patients to 42 months. Five patients got complete remission and 1 patient got partial remission at 1 month after implantation. The objective response rate was 100%. The median survival time was 26 months. And median progression-free survival was 12 months after ¹²⁵I treatment. And about the complications, 1 patient suffered from the light aerothorax, 1 patient had a little hemoptysis. Our study showed that ¹²⁵I seed implantation as second line regimen in small cell lung cancer that recurred locally after first line treatment was effective and safe. That could improve the overall survival and progression-free survival only comparing to the second line chemotherapy. Therefore ¹²⁵I seed implantation as brachytherapy protocol is a promising method and can be applied as second line treatment to control the locally recurred small cell lung cancer.     

Local maternal control of seed size by KLUH/CYP78A5-dependent growth signaling

Seed development in plants involves the coordinated growth of the embryo, endosperm, and maternal tissue. Several genes have been identified that influence seed size by acting maternally, such as AUXIN RESPONSE FACTOR2, APETALA2, and DA1. However, given the lack of gain-of-function effects of these genes on seed size, it is unclear whether their activity levels are limiting in WT plants and whether they could thus be used to regulate seed size in development or evolution. Also, whether the altered seed sizes reflect local gene activity or global physiological changes is unknown. Here, we demonstrate that the cytochrome P450 KLUH (KLU) regulates seed size. KLU acts locally in developing flowers to promote seed growth, and its activity level is limiting for seed growth in WT. KLU is expressed in the inner integument of developing ovules, where it non-cell autonomously stimulates cell proliferation, thus determining the growth potential of the seed coat and seed. A KLU-induced increase in seed size leads to larger seedlings and higher relative oil content of the seeds. Genetic analyses indicate that KLU acts independently of other tested maternal factors that influence integument cell proliferation. Thus, the level of KLU-dependent growth factor signaling determines size in ovules and seeds, suggesting this pathway as a target for crop improvement.     

Evidence for participation of the methionine sulfoxide reductase repair system in plant seed longevity

Seeds are in a natural oxidative context leading to protein oxidation. Although inevitable for proper progression from maturation to germination, protein oxidation at high levels is detrimental and associated with seed aging. Oxidation of methionine to methionine sulfoxide is a common form of damage observed during aging in all organisms. This damage is reversible through the action of methionine sulfoxide reductases (MSRs), which play key roles in lifespan control in yeast and animal cells. To investigate the relationship between MSR capacity and longevity in plant seeds, we first used two Medicago truncatula genotypes with contrasting seed quality. After characterizing the MSR family in this species, we analyzed gene expression and enzymatic activity in immature and mature seeds exhibiting distinct quality levels. We found a very strong correlation between the initial MSR capacities in different lots of mature seeds of the two genotypes and the time to a drop in viability to 50% after controlled deterioration. We then analyzed seed longevity in Arabidopsis thaliana lines, in which MSR gene expression has been genetically altered, and observed a positive correlation between MSR capacity and longevity in these seeds as well. Based on our data, we propose that the MSR repair system plays a decisive role in the establishment and preservation of longevity in plant seeds.     

Intraoperative radiofrequency ablation combined with 125 iodine seed implantation for unresectable pancreatic cancer

AIM: To evaluate the feasibility, efficacy and safety of intraoperative radiofrequency ablation (RFA) combined with 125 iodine seed implantation for unresectable pancreatic cancer. METHODS: Thirty-two patients (21 males and 11 females) at the age of 68 years (range 48-90 years) with unresectable locally advanced pancreatic cancer admitted to our hospital from January 2006 to May 2008 were enrolled in this study. The tumor, 4-12 cm in diameter, located in pancreatic head of 23 patients and in pancreatic body...     

Plastochromanol-8 and tocopherols are essential lipid-soluble antioxidants during seed desiccation and quiescence in Arabidopsis

Given their essential role as vitamin E, tocopherols and tocotrienols have been studied extensively in animals and plants. In contrast, our understanding of the function of plastochromanol-8 (PC-8), a third type of tocochromanol with a longer side chain, is very limited despite the wide distribution of PC-8 in the plant kingdom, including species consumed by humans. To investigate PC-8 function in vivo, we combined the Arabidopsis vte1 mutation that eliminates tocopherols and PC-8 and causes the accumulation of 2,3-dimethyl-6-phytyl-1,4-benzoquinol (DMPBQ), a redox-active tocopherol precursor, and the vte2 mutation that eliminates tocopherols without affecting PC-8. The vte2 vte1 double mutant lacks tocopherols, PC-8, and DMPBQ, and exhibits the most severe physiological and biochemical phenotypes of any tocochromanol-affected genotype isolated to date, most notably a severe seedling developmental phenotype associated with massive lipid oxidation initiated during seed desiccation and amplified during seed quiescence. In contrast, the presence of PC-8 in vte2 suppresses or attenuates all of the developmental and biochemical phenotypes observed in vte2 vte1, demonstrating that PC-8 is a lipid antioxidant in vivo. Finally, the low relative fitness of vte2 vte1 demonstrates that tocopherols and PC-8 are in vivo lipid antioxidants essential for seed plant survival.     

The Role of Tissue Resident Memory CD4 T Cells in Herpes Simplex Viral and HIV Infection

Tissue-resident memory T cells (TRM) were first described in 2009. While initially the major focus was on CD8⁺ TRM, there has recently been increased interest in defining the phenotype and the role of CD4⁺ TRM in diseases. Circulating CD4⁺ T cells seed CD4⁺ TRM, but there also appears to be an equilibrium between CD4⁺ TRM and blood CD4⁺ T cells. CD4⁺ TRM are more mobile than CD8⁺ TRM, usually localized deeper within the dermis/lamina propria and yet may exhibit synergy with CD8⁺ TRM in disease control. This has been demonstrated in herpes simplex infections in mice. In human recurrent herpes infections, both CD4⁺ and CD8⁺ TRM persisting between lesions may control asymptomatic shedding through interferon-gamma secretion, although this has been more clearly shown for CD8⁺ T cells. The exact role of the CD4⁺/CD8⁺ TRM axis in the trigeminal ganglia and/or cornea in controlling recurrent herpetic keratitis is unknown. In HIV, CD4⁺ TRM have now been shown to be a major target for productive and latent infection in the cervix. In HSV and HIV co-infections, CD4⁺ TRM persisting in the dermis support HIV replication. Further understanding of the role of CD4⁺ TRM and their induction by vaccines may help control sexual transmission by both viruses.