Monokine Induced by Interferon-γ (MIG/CXCL9) Is Derived from Both Donor and Recipient Sources during Rejection of Class II Major Histocompatibility Complex Disparate Skin Allografts

Chemokines, including monokine induced by interferon-γ (Mig/CXCL9), are produced both in allografts and during the direct T-cell infiltration that mediates graft rejection. Neither the specific production nor contribution of allograft donor versus recipient Mig in allograft rejection is currently known. C57BL/6 mice with a targeted deletion in the Mig gene were used as both skin allograft donors and recipients in a class II major histocompatibility complex-mismatched graft model to test the requirement for donor- versus recipient-derived Mig for acute rejection. B6.Mig^−/− allografts had a 10-day prolonged survival in B6.H-2^bm12 recipients when compared with wild-type C57BL/6 allograft donors, and B6.H-2^bm12 skin allografts had a 5-day prolonged survival in B6.Mig^−/− versus wild-type recipients. Transplantation of B6.Mig^−/− skin grafts onto B6.H-2^bm12.Mig^−/− recipients resulted in further prolonged allograft survival with more than 30% of the grafts surviving longer than 60 days. Prolonged allograft survival was also associated with delayed cellular infiltration into grafts but not with altered T-cell proliferative responses to donor stimulators. Immunohistochemical staining of allograft sections indicated that Mig is produced by both donor- and recipient-derived sources, but Mig from each of these sources appeared in different areas of the allograft tissue. These results therefore demonstrate the synergy of donor- and recipient-derived Mig in promoting T-cell infiltration into allografts.Acute allograft rejection is mediated by the coordinated infiltration of alloantigen-primed T cells into the graft and the expression of effector functions that destroy the vascular endothelium and the parenchymal tissue.^1,2 Adhesion molecules and chemoattractant cytokines, chemokines, play major roles in directing primed T-cell recruitment and infiltration into allografts.^3,4,5 The role of adhesion molecules in graft rejection is indicated by the ability of specific antibodies or the use of adhesion molecule-deficient graft recipients or donors to delay or inhibit acute allograft rejection in many animal models.^6,7,8 Similarly, many studies have demonstrated the ability to delay or inhibit allograft rejection through administration of antibodies to specific chemokines or chemokine receptors.⁹ In addition, the use of graft recipients with targeted deletions in CXCR3 and CCR5 has supported a role for these receptors in promoting T-cell trafficking to mediate acute rejection.^10,11 Although these studies indicate an important function of specific chemokines in directing T-cell infiltration into allografts, the induction and source of these chemokines during the rejection process remains poorly understood.The CXCR3 ligands, Mig/CXCL9, IP-10/CXCL10, and I-TAC/CXCL11, are potent chemoattractants for antigen-activated T cells.^12,13 These chemokines are induced by interferon (IFN)-γ and are produced during many T-cell-mediated inflammatory responses including allograft rejection. Mig/CXCL9 is produced at low levels in skin and heart allografts early after transplantation in mouse models but this production increases with alloantigen-primed T-cell infiltration and activity in the allograft.^14,15 Consistent with animal models, the expression of Mig in biopsies from clinical renal and heart allografts is indicative of an ongoing acute rejection episode.^16,17 In rodent models, treatment with Mig-specific antibodies delays T-cell infiltration and prolongs the survival of complete major histocompatibility complex (MHC)-mismatched skin allografts 3 to 5 days implicating a role for Mig in optimal T-cell recruitment into grafts.¹⁸ This is supported by the ability of chronic treatment of C57BL/6 recipients with Mig-specific antibodies to promote the survival of ∼75% of single class II MHC-disparate B6.H-2^bm12 full thickness trunk skin allografts until the treatment is stopped.¹⁹Mig is produced by endothelial cells and macrophages during many inflammatory processes.²⁰ The production of Mig by donor- and recipient-derived sources during allograft rejection remains unclear and the relative contribution of each source in allograft rejection is untested. In the current study, we have used mice with a targeted deletion in the Mig gene as allograft donors and recipients to test these aspects of the skin allograft rejection process. The results indicate the production of Mig by both graft- and recipient-derived sources but the production of each source appears in different tissue locations and affects the time of T-cell graft infiltration during the acute rejection process.     

Comparison of mechanical and electrical activity and interstitial cells of Cajal in urinary bladders from wild-type and W/Wv mice

Background and purpose:

W/W^v and wild-type murine bladders were studied to determine whether the W/W^v phenotype, which causes a reduction in, but not abolition of, tyrosine kinase activity, is a useful tool to study the function of bladder interstitial cells of Cajal (ICC).

Experimental approach:

Immunohistochemistry, tension recordings and microelectrode recordings of membrane potential were performed on wild-type and mutant bladders.

Key results:

Wild-type and W/W^v detrusors contained c-Kit- and vimentin-immunopositive cells in comparable quantities, distribution and morphology. Electrical field stimulation evoked tetrodotoxin-sensitive contractions in wild-type and W/W^v detrusor strips. Atropine reduced wild-type responses by 50% whereas a 25% reduction occurred in W/W^v strips. The atropine-insensitive component was blocked by pyridoxal-5-phosphate-6-azophenyl-2′,4′-disulphonic acid in both tissue types. Wild-type and W/W^v detrusors had similar resting membrane potentials of −48 mV. Spontaneous electrical activity in both tissue types comprised action potentials and unitary potentials. Action potentials were nifedipine-sensitive whereas unitary potentials were not. Excitatory junction potentials were evoked by single pulses in both tissues. These were reduced by atropine in wild-type tissues but not in W/W^v preparations. The atropine-insensitive component was abolished by pyridoxal-5-phosphate-6-azophenyl-2′,4′-disulphonic acid in both preparations.

Conclusions and implications:

Bladders from W/W^v mice contain c-Kit- and vimentin-immunopositive ICC. There are similarities in the electrical and contractile properties of W/W^v and wild-type detrusors. However, significant differences were found in the pharmacology of the responses to neurogenic stimulation with an apparent up-regulation of the purinergic component. These findings indicate that the W/W^v strain may not be the best model to study ICC function in the bladder.     

An Ecological Risk Assessment of the Exposure and Effects of 2,4-D Acid to Rainbow Trout (<Emphasis Type="Italic">Onchorhyncus mykiss</Emphasis>)

Numerous state and federal agencies are increasingly concerned with the rapid expansion of invasive, noxious weeds across the United States. Herbicides are frequently applied as weed control measures in forest and rangeland ecosystems that frequently overlap with critical habitats of threatened and endangered fish species. However, there is little published chronic toxicity data for herbicides and fish that can be used to assess ecological risk of herbicides in aquatic environments. We conducted 96-h flowthrough acute and 30-day chronic toxicity studies with swim-up larvae and juvenile rainbow trout (Onchorhyncus mykiss) exposed to the free acid form of 2,4-D. Juvenile rainbow trout were acutely sensitive to 2,4-D acid equivalent at 494 mg/L (95% confidence interval [CI] 334–668 mg/L; 96-h ALC₅₀). Accelerated life-testing procedures, used to estimate chronic mortality from acute data, predicted that a 30-day exposure of juvenile rainbow trout to 2,4-D would result in 1% and 10% mortality at 260 and 343 mg/L, respectively. Swim-up larvae were chronically more sensitive than juveniles using growth as the measurement end point. The 30-day lowest observable effect concentration (LOEC) of 2,4-D on growth of swim-up larvae was 108 mg/L, whereas the 30-day no observable effect concentration (NOEC) was 54 mg/L. The 30-day maximum acceptable toxicant concentration (MATC) of 2,4-D for rainbow trout, determined as the geometric mean of the NOEC and the LOEC, was 76 mg/L. The acute:chronic ratio was 6.5 (i.e., 494/76). We observed no chronic effects on growth of juvenile rainbow trout at the highest concentration tested (108 mg/L). Worst-case aquatic exposures to 2,4-D (4 mg/L) occur when the herbicide is directly applied to aquatic ecosystems for aquatic weed control and resulted in a 30-day safety factor of 19 based on the MATC for growth (i.e., 76/4). Highest nontarget aquatic exposures to 2,4-D applied following terrestrial use is calculated at 0.136 mg/L and resulted in a 30-day safety factor of 559 (e.g., 76/0.163). Assessment of the exposure and response data presented herein indicates that use of 2,4-D acid for invasive weed control in aquatic and terrestrial habitats poses no substantial risk to growth or survival of rainbow trout or other salmonids, including the threatened bull trout (Salvelinus confluentus).     

Ecological effects of lead mining on Ozark streams: In-situ toxicity to woodland crayfish (Orconectes hylas)

The Viburnum Trend mining district in southeast Missouri, USA is one of the largest producers of lead–zinc ore in the world. Previous stream surveys found evidence of increased metal exposure and reduced population densities of crayfish immediately downstream of mining sites. We conducted an in-situ 28-d exposure to assess toxicity of mining-derived metals to the woodland crayfish (Orconectes hylas). Crayfish survival and biomass were significantly lower at mining sites than at reference and downstream sites. Metal concentrations in water, detritus, macroinvertebrates, fish, and crayfish were significantly higher at mining sites, and were negatively correlated with caged crayfish survival. These results support previous field and laboratory studies that showed mining-derived metals negatively affect O. hylas populations in streams draining the Viburnum Trend, and that in-situ toxicity testing was a valuable tool for assessing the impacts of mining on crayfish populations.     

Design,analysis, and interpretation of treatment response heterogeneity in personalized nutrition and obesity treatment research

It is increasingly assumed that there is no one-size-fits-all approach to dietary recommendations for the management and treatment of chronic diseases such as obesity. This phenomenon that not all individuals respond uniformly to a given treatment has become an area of research interest given the rise of personalized and precision medicine. To conduct, interpret, and disseminate this research rigorously and with scientific accuracy, however, requires an understanding of treatment response heterogeneity. Here, we define treatment response heterogeneity as it relates to clinical trials, provide statistical guidance for measuring treatment response heterogeneity, and highlight study designs that can quantify treatment response heterogeneity in nutrition and obesity research. Our goal is to educate nutrition and obesity researchers in how to correctly identify and consider treatment response heterogeneity when analyzing data and interpreting results, leading to rigorous and accurate advancements in the field of personalized medicine.     

Predictors of successful labor induction with oral or vaginal misoprostol

Objective: To identify independent predictors of successful labor induction with oral or vaginal misoprostol.

Methods: Women enrolled in four previous randomized trials involving oral or vaginal misoprostol for cervical ripening and labor induction were included in the present cohort study, with dosing of 25–50?μg every 4 to 6?h vaginally (n?=?574) or 50?μg every 4?h orally (n?=?207). Multiple logistic regression was performed to identify factors independently associated with successful labor induction – defined as vaginal delivery within 12?h, vaginal delivery within 24?h and spontaneous vaginal delivery. Predictors of Cesarean birth and the need for only one dose of misoprostol were also identified. Variables included in the models were maternal age, weight, height, parity, gravidity, membrane status, route of misoprostol, gestational age, birth weight, and Bishop score and its individual components.

Results: Maternal age, height, weight, parity, birth weight, dilatation, effacement and cervical station were associated with vaginal delivery within 24?h of induction. Maternal age, height, weight, nulliparity, birth weight and route of misoprostol were associated with Cesarean birth, with oral misoprostol being associated with a lower rate of Cesarean birth. The need for only one dose of misoprostol was predicted by maternal height, weight, parity, gestational age, Bishop score and route of misoprostol.

Conclusion: Characteristics of the woman (height, weight, parity), the fetus (birth weight) and some of the individual components of the Bishop score, were associated with successful labor induction, with oral misoprostol being associated with a lower rate of Cesarean birth.     

Endogenous ligands selecting T cells expressing particular V beta elements.

It has recently become clear that the minor lymphocyte stimulatory antigens (Mls) and other endogenous ligands which lead to the partial or total deletion of T cells bearing particular V beta segments are encoded by mouse mammary tumor virus (MMTV). We review here the genetic analyses of multiple V beta 11 and V beta 3 deletion ligands and demonstrate the involvement of MMTV in all examples. Several features of Mls and the V beta 11/V beta 3 deleting ligands identify them as members of the superantigen family. Bacterial superantigens are known to bind both MHC class II and the TCR in regions distinct from conventional peptide antigens. Within the MMTV genome, the 3' LTR has been identified as encoding superantigen function. We present data demonstrating that in vitro translation identifies the major product of the open reading frame (ORF) within the 3' LTR as a type II integral membrane glycoprotein. It is proposed that the type II membrane glycoprotein interacts with MHC and TCR in a manner analogous to the bacterial superantigens and distinct from conventional peptide antigen. Several unanswered questions regarding superantigen action remain; what determines total or partial deletion? How is Mls transferred between cells? These questions are addressed in the discussion.     

人脂肪组织来源干细胞在心脏疾病治疗中的应用

目的:综述人脂肪组织来源干细胞的生物学特性及其在缺血性心脏病中的应用,分析不足,并在此基础上提出未来研究要解决的问题,以期为临床治疗提供依据。资料来源:应用计算机检索Blackwell、Elsevier、Pubmed数据库1980/2007期间脂肪源性干细胞与缺血性心脏病方面的文献,检索词为“bone mesenchymal stem cells,adipose derived stemcells,cardiomyocytes,ischemic heart disease”等。应用计算机检索中国期刊全文数据库1980/2007期间相关文献,检索词为“骨髓间充质干细胞,脂肪组织来源的干细胞,心肌细胞,缺血性心脏病”等。并手工查阅相关书籍。资料选择:对资料进行初步选择:①脂肪组织来源干细胞的生物学特性。②脂肪组织来源干细胞治疗缺血性心脏病。排除重复文献。资料提炼:共搜集到相关文章57篇,删除内容重复及与本文主题关系较远的文章,剩余41篇作为综述参考。资料综合:脂肪组织来源干细胞与同样起源于中胚层的骨髓基质细胞不仅具有非常相似的生物学特性,而且在细胞表面标志谱的表达方面也非常相近。并且脂肪组织来源广泛,取材方便,可获得的基质细胞数量大,易于培养扩增。有研究发现,脂肪组织来源干细胞体外培养不需要任何诱导便能分化成具有自律性的心肌细胞,使得脂肪组织来源干细胞治疗缺血性心脏病成为可能。结论:脂肪组织来源干细胞在取材和增殖方面较骨髓间充质干细胞有优势;脂肪组织来源干细胞能较好的诱导为心肌细胞,将为缺血性心脏病的治疗提供更广阔的前景。