Probiotics and synbiotics for preventing postoperative infectious complications in colorectal cancer patients: a systematic review and meta-analysis.

BACKGROUND: The health benefits of probiotics and synbiotics in healthy adults are well established, but their role in preventing infectious complications after surgery for colorectal cancer remains controversial. The aim of this meta-analysis was to assess the impact of probiotics/synbiotics on the incidence of infectious complications in patients who had surgery for colorectal cancer. METHODS: A comprehensive literature search of all randomized control trials (RCTs) was conducted using PubMed, Embase, World Health Organization (WHO) Global Index Medicus, WHO clinical trial registry, and Clinicaltrials.gov. Inclusion criteria included RCTs comparing the use of any strain or dose of a specified probiotic/synbiotic with placebo or a "standard care" control group. The incidence of postoperative infectious complications was analyzed. RESULTS: Fourteen RCTs involving 1566 patients (502 receiving probiotics, 273 receiving synbiotics, and 791 receiving placebo) were analyzed. Overall, probiotic or synbiotic administration significantly reduced the risk of developing postoperative infectious complications by 37% (relative risk (RR) = 0.63, 95% confidence interval (CI) 0.54-0.74, p < 0.001). Furthermore, when considering the six different types of postoperative infectious complications (septicemia, incision infection, central line infection, pneumonia infection, urinary infection, and incidence of diarrhea), probiotic or synbiotic administration was beneficial in reducing the incidence of each one of them. The quality of evidence was listed below: incidence of diarrhea (high), septicemia (moderate), incision infection (moderate), pneumonia infection (moderate), urinary infection (moderate), and central line infection (low). However, for the main outcome of infectious complications, we found evidence of possible publication bias, although estimates still showed a reduction following trim-and-fill analysis (RR = 0.72, 95% CI 0.62-0.84, p < 0.001). CONCLUSIONS: The use of probiotic/synbiotic supplementation is associated with a significant reduction in the risk of developing postoperative infectious complications in patients who had surgery for colorectal cancer. Additional studies are needed to confirm the findings due to publication bias and low quality of evidence.

[Predicting postoperative recurrence of stage â -â ¢ renal clear cell carcinoma based on preoperative CT radiomics feature nomogram].

OBJECTIVE: To explore the preoperative radiomics features (RFs) and construct a nomogram for predicting postoperative recurrence of stage Ⅰ-Ⅲ clear cell renal carcinoma (ccRCC). METHODS: The clinicopathological data and preoperative enhanced CT images collected from 256 patients with ccRCC were used as the training dataset (175 patients) and test dataset (81 patients). The enhanced CT images of the tumor were segmented using ITK-SNAP software, and the RFs were extracted using the PyRadiomics computing platform. In the training dataset, the RFs were screened based on Lasso-CV algorithm, and the Rad_score was calculated. The Clinic factors were screened by univariate and multivariate logistic regression analysis of the clinical and pathological factors and CT characteristics. The Rad_score, Clinic、Rad_score + Clinic nomograms were constructed and verified using the test dataset. The performance, discrimination power and calibration of the nomograms were compared, and their clinical value was evaluated using decision curve analysis. RESULTS: Six RFs were retained to calculate the Rad_score. The Clinic factors included Rad_score, KPS score, platelet, calcification and TNM clinical stage. In terms of discrimination, the Rad_score + Clinic nomogram showed better performance (AUC=0.84 for training set; AUC=0.85 for test set) than the Rad_score nomogram (AUC=0.78 for training set, P=0.029; AUC=0.77 for Test set, P=0.025) and Clinic nomogram (AUC=0.77 for training set, P=0.014; AUC=0.77 for test set, P=0.011). In terms of calibration, the P value for goodness of fit test of the Rad_score+Clinic nomogram was 0.065 for the training set and 0.628 for the test set. Decision curve analysis showed a greater clinical value of the Rad_score+Clinic nomogram with Rad_score than the Clinic nomogram without Rad_score. CONCLUSION: The nomogram based on preoperative CT RFs has a high value for predicting postoperative recurrence of stage Ⅰ-Ⅲ ccRCC to facilitate individualized treatment of RCC.

Syphilis-related rapidly progressive glomerulonephritis: a case presentation.

BACKGROUND: Syphilis is a multisystemic infection that causes a wide variety of symptoms and thus has been dubbed one of the great medical mimickers. Due to recent global re-emergence of syphilis, it has become important to recognize its various presentations. Relative to the kidney, syphilitic infections generally present themselves with nephrotic range proteinuria, and are most often associated with pathological features of a membranous glomerulonephritis with subepithelial immune complex deposition. However, other rare renal presentations have been reported. One of these includes a rapidly progressive glomerulonephritis picture. All described cases have been successfully resolved with the treatment of the underlying syphilis infection. CASE PRESENTATION: The patient was an elderly woman of Caribbean descent who presented with lower extremity weakness, anasarca and proteinuria, hematuria with progressive renal failure. On kidney biopsy, she was found to have a pauci-immune crescentic glomerulonephritis pattern and a concomitant acute tubulointerstitial nephritis. She had a positive Treponema pallidum particle agglutination test and a negative syphilis rapid plasma reagin test with clinical evidence of polyneuropathy suggestive chronic syphilis infection. CONCLUSION AND DISCUSSION: It is important in the context of pauci-immune crescentic glomerulonephritis to explore all differential diagnoses. Given the positive syphilis serologies, clinical context and presence of tubulointerstitial nephritis, she was determined to have syphilitic glomerulonephritis that resolved with a course of both penicillin and steroids.

[Comparative study on changes of ginsenosides and activities of American ginseng before and after steaming].

The chemical fingerprints of American ginseng were established by high performance liquid chromatography(HPLC) coupled with similarity evaluation system for chromatographic fingerprint of traditionanl Chinese medicine. The results were analyzed with use of stoichiometry methods(cluster analysis, principal component analysis, and orthogonal partial least squares discriminant analysis), and meanwhile, a preliminary study on the antioxidant and anti-proliferation activity of colorectal cancer cells was conducted. By comparing the fingerprints of American ginseng before and after processing, the contents of five components in the eight ginseno-sides quantified in this paper increased, including ginsenoside Rc, Rg_2, Rb_2, Rb_3 and Rd, respectively, and a new component was produced after steaming. The activity study showed that steamed American ginseng had better antioxidant activity and anti-proliferation activity of colorectal cancer cells than raw American ginseng. The research results show that the steaming method of American ginseng used in this experiment has good stability and reproducibility, and the steaming of American ginseng produces similar changes as artificial red ginseng, which provides a certain reference for expanding the application range of American ginseng.

Evaluating Inoculation Methods to Infect Sugar Beet with Fusarium oxysporum f. betae and F. secorum.

Minnesota and North Dakota combined contain 55% of the sugar beet production area in the United States, contributing to 49% of the nation's sugar beet production in 2018. Fusarium diseases caused by Fusarium oxysporum f. betae and F. secorum on sugar beet can cause significant reduction in both root yield and sucrose concentration and purity. The objective of this research was to identify an alternative artificial inoculation method to induce Fusarium diseases on sugar beet leaves and roots caused by both Fusarium spp. in greenhouse conditions to better aid in research efforts. We tested four inoculation methods, including barley to seed, barley to root, drenching, and cutting. and compared them with the conventional root-dipping inoculation method. The inoculation method of placing Fusarium-colonized barley seed close to sugar beet seed (barley to seed) caused levels of symptom severities on both leaves and roots similar to the root-dipping method. Because the traditional root-dipping method involves a laborious transplant process, use of infected barley seed as inoculum may serve as an alternative method in the evaluation of host resistance and pathogen virulence among Fusarium diseases by Fusarium spp. on sugar beet at the seed or seedling stage.

Computational analysis of alternative splicing in plant genomes.

Computational analyses play crucial roles in characterizing splicing isoforms in plant genomes. In this review, we provide a survey of computational tools used in recently published, genome-scale splicing analyses in plants. We summarize the commonly used software and pipelines for read mapping, isoform reconstruction, isoform quantification, and differential expression analysis. We also discuss methods for analyzing long reads and the strategies to combine long and short reads in identifying splicing isoforms. We review several tools for characterizing local splicing events, splicing graphs, coding potential, and visualizing splicing isoforms. We further discuss the procedures for identifying conserved splicing isoforms across plant species. Finally, we discuss the outlook of integrating other genomic data with splicing analyses to identify regulatory mechanisms of AS on genome-wide scale.

Effectiveness of Rlm7 resistance against Leptosphaeria maculans (phoma stem canker) in UK winter oilseed rape cultivars.

The Rlm7 gene in Brassica napus is an important source of resistance for control of phoma stem canker on oilseed rape caused by the fungus Leptosphaeria maculans. This study shows the first report of L. maculans isolates virulent against Rlm7 in the UK. Leptosphaeria maculans isolates virulent against Rlm7 represented 3% of the pathogen population when cultivars with the Rlm7 gene represented 5% of the UK oilseed rape area in 2012/13. However, the Rlm7 gene has been widely used since then, representing >15% of the UK oilseed rape area in 2015/16. Winter oilseed rape field experiments included cultivars with the Rlm7 gene, with the Rlm4 gene or without Rlm genes and took place at five sites in the UK over four cropping seasons. An increase in phoma leaf spotting severity on Rlm7 cultivars in successive seasons was observed. Major resistance genes played a role in preventing severe phoma leaf spotting at the beginning of the cropping season and, in addition, quantitative resistance (QR) in the cultivars examined made an important contribution to control of phoma stem canker development at the end of the cropping season. Deployment of the Rlm7 resistance gene against L. maculans in cultivars with QR in combination with sustainable disease management practices will prolong the use of this gene for effective control of phoma stem canker epidemics.

Genetic diversity and population structure of Kazakh horses (Equus caballus) inferred from mtDNA sequences.

The Kazakh horse is an important old horse breed in Xinjiang. They have contributed greatly to the breeding and improvement of other local horse breeds, yet their genetic diversity and population structure are not well understood. In the present study, we evaluated the genetic diversity of Kazakh horses and their relationship with other horse breeds using the mtDNA D-loop region, Cyt b gene, and a DNA fragment (nps 7974-9963, containing COX3, tRNA-Gly, ND3, and tRNA-Arg). A total of 130 Kazakh horses from 8 populations in China and Kazakhstan were analyzed. A total of 88 haplotypes (haplotype diversity: 0.9895) were identified, in which 3 haplotypes were shared by groups in the two countries. In a median-joining network, 6 haplogroups were found, in which most haplogroups included haplotypes from different populations. Neighbor-joining analysis revealed similar results in that haplotypes in different populations were admixed in most of the 6 clusters. In conclusion, a high level of genetic diversity was found in the Kazakh horses. However, no clear correspondence between haplogroups and geographic origin and no significant differentiation between populations in the two countries were observed. This might have resulted from the frequent contact between the two countries through the Silk Road in the past, or due to long-term outcrossing and hybridization with the introduced horses.

Negative emotions affect postoperative scores for evaluating functional knee recovery and quality of life after total knee replacement.

This study aimed to determine whether psychological factors affect health-related quality of life (HRQL) and recovery of knee function in total knee replacement (TKR) patients. A total of 119 TKR patients (male: 38; female: 81) completed the Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI), State Trait Anxiety Inventory (STAI), Eysenck Personality Questionnaire-revised (EPQR-S), Knee Society Score (KSS), and HRQL (SF-36). At 1 and 6 months after surgery, anxiety, depression, and KSS scores in TKR patients were significantly better compared with those preoperatively (P<0.05). SF-36 scores at the sixth month after surgery were significantly improved compared with preoperative scores (P<0.001). Preoperative Physical Component Summary Scale (PCS) and Mental Component Summary Scale (MCS) scores were negatively associated with extraversion (E score) (B=-0.986 and -0.967, respectively, both P<0.05). Postoperative PCS and State Anxiety Inventory (SAI) scores were negatively associated with neuroticism (N score; B=-0.137 and -0.991, respectively, both P<0.05). Postoperative MCS, SAI, Trait Anxiety Inventory (TAI), and BAI scores were also negatively associated with the N score (B=-0.367, -0.107, -0.281, and -0.851, respectively, all P<0.05). The KSS function score at the sixth month after surgery was negatively associated with TAI and N scores (B=-0.315 and -0.532, respectively, both P<0.05), but positively associated with the E score (B=0.215, P<0.05). The postoperative KSS joint score was positively associated with postoperative PCS (B=0.356, P<0.05). In conclusion, for TKR patients, the scores used for evaluating recovery of knee function and HRQL after 6 months are inversely associated with the presence of negative emotions.

Negative emotions affect postoperative scores for evaluating functional knee recovery and quality of life after total knee replacement

This study aimed to determine whether psychological factors affect health-related quality of life (HRQL) and recovery of knee function in total knee replacement (TKR) patients. A total of 119 TKR patients (male: 38; female: 81) completed the Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI), State Trait Anxiety Inventory (STAI), Eysenck Personality Questionnaire-revised (EPQR-S), Knee Society Score (KSS), and HRQL (SF-36). At 1 and 6 months after surgery, anxiety, depression, and KSS scores in TKR patients were significantly better compared with those preoperatively (P<0.05). SF-36 scores at the sixth month after surgery were significantly improved compared with preoperative scores (P<0.001). Preoperative Physical Component Summary Scale (PCS) and Mental Component Summary Scale (MCS) scores were negatively associated with extraversion (E score) (B=-0.986 and -0.967, respectively, both P<0.05). Postoperative PCS and State Anxiety Inventory (SAI) scores were negatively associated with neuroticism (N score; B=-0.137 and -0.991, respectively, both P<0.05). Postoperative MCS, SAI, Trait Anxiety Inventory (TAI), and BAI scores were also negatively associated with the N score (B=-0.367, -0.107, -0.281, and -0.851, respectively, all P<0.05). The KSS function score at the sixth month after surgery was negatively associated with TAI and N scores (B=-0.315 and -0.532, respectively, both P<0.05), but positively associated with the E score (B=0.215, P<0.05). The postoperative KSS joint score was positively associated with postoperative PCS (B=0.356, P<0.05). In conclusion, for TKR patients, the scores used for evaluating recovery of knee function and HRQL after 6 months are inversely associated with the presence of negative emotions.

Calcium and cholecalciferol supplementation provides no added benefit to nutritional counseling to improve bone mineral density in survivors of childhood acute lymphoblastic leukemia (ALL).

BACKGROUND: We sought to improve lumbar spine bone mineral density (LS-BMD) in long-term survivors of childhood acute lymphoblastic leukemia (ALL) using calcium and cholecalciferol supplementation. PROCEDURE: This double-blind, placebo-controlled trial randomized 275 participants (median age, 17 [9-36.1] years) with age- and gender-specific LS-BMD Z-scores <0 to receive nutritional counseling with supplementation of 1,000 mg/day calcium and 800 International Unit cholecalciferol or placebo for 2 years. The primary outcome was change in LS-BMD assessed by quantitative computerized tomography (QCT) at 24 months. Linear regression models were employed to identify the baseline risk factors for low LS-BMD and to compare LS-BMD outcomes. RESULTS: Pre-randomization LS-BMD below the mean was associated with male gender (P = 0.0024), White race (P = 0.0003), lower body mass index (P < 0.0001), and cumulative glucocorticoid doses of ≥ 5,000 mg (P = 0.0012). One hundred eighty-eight (68%) participants completed the study; 77% adhered to the intervention. Mean LS-BMD change did not differ between survivors randomized to supplements (0.33 ± 0.57) or placebo (0.28 ± 0.56). Participants aged 9-13 years and those 22-35 years had the greatest mean increases in LS-BMD (0.50 ± 0.66 and 0.37 ± 0.23, respectively). Vitamin D insufficiency (serum 25[OH]D <30 ng/ml) found in 296 (75%), was not associated with LS-BMD outcomes (P = 0.78). CONCLUSION: Cholecalciferol and calcium supplementation provides no added benefit to nutritional counseling for improving LS-BMD among adolescent and young adult survivors of ALL (93% of whom had LS-BMD Z-scores above the mean at study entry).

Propofol prevents cerebral ischemia-triggered autophagy activation and cell death in the rat hippocampus through the NF-κB/p53 signaling pathway.

Propofol (2,6-diisopropylphenol) has been shown to attenuate neuronal injury under a number of experimental conditions; however, the mechanisms involved in its neuroprotective effects remain unclear. We therefore investigated whether inhibition of p53 induction by propofol contributes to the neuroprotection of cerebral ischemic cell death through both autophagic and apoptotic mechanisms. A transient global cerebral ischemia-reperfusion (I/R) model was produced with a 10-min, 2-vessel occlusion. The change in target genes including damage-regulated autophagy modulator (DRAM), microtubule-associated protein 1 light chain 3 (LC3), Beclin 1, cathepsin D, cathepsin B, p53-upregulated modulator of apoptosis (PUMA), Bax and Bcl-2 upon p53 inhibition was assessed with the co-administration of the intravenous anesthetic propofol and 3-methyladenine (3-MA), Pifithrin-alpha (PFT-α) or SN50. The I/R-induced increases of protein levels of p53 and LC3-II were significantly inhibited by treatment with propofol, 3-MA or PFT-α. The I/R-induced increases of protein levels of DRAM, Beclin 1, active cathepsin D and cathepsin B were significantly inhibited by treatment with propofol, PFT-α or SN50. The negative effects of the I/R-induced up-regulation of PUMA and Bax and the down-regulation of Bcl-2 in the rat hippocampus were all blocked by treatment with propofol, PFT-α or SN50. Our results suggest that cerebral I/R can induce nuclear factor-kappa B-dependent expression of p53. The autophagic and apoptotic mechanisms participate in programed cell death by regulating the p53-mediated pathway. Our results are the first to show that propofol, at clinically relevant concentrations, attenuated cell death through both autophagic and apoptotic mechanisms in the rat hippocampus after a cerebral I/R insult.

Identification of blood protein biomarkers that aid in the clinical assessment of patients with malignant glioma.

Analyzing molecular biomarkers using blood is an important approach for clinical assessment of malignant glioma. We investigated a molecular proteomic biomarker-based approach for glioblastoma using patients' blood samples. The expression levels of a list of candidate proteins were quantified in plasma and serum samples from two different cohorts of patients with malignant glioma and normal controls. The biological function was studied for one of the identified markers. Additionally, the prognostic significance of protein marker expression was measured by survival analysis. As a result, protein biomarkers associated with malignant glioma were identified from the blood specimens and five of the protein biomarkers were common to both cohorts. Immunohistochemical analysis demonstrated that many of the protein biomarkers identified in peripheral blood specimens were expressed in malignant gliomas. Staining levels for one of the biomarkers, MIP-1α, was found to correlate with WHO grade among invasive gliomas, and we demonstrate that MIP-1α promotes human glioblastoma cell proliferation and migration. Additionally, four prognostic protein biomarkers were identified. In conclusion, we demonstrate that both peripheral blood plasma and serum specimens are highly valuable and complementary to each other in the quest for protein biomarkers of malignant glioma. Sets of novel protein biomarkers were identified that may aid in the diagnosis and prognosis of patients with malignant glioma.

Comparative Pathogenicity and Virulence of Fusarium Species on Sugar Beet.

In all, 98 isolates of three Fusarium spp. (18 Fusarium oxysporum, 30 F. graminearum, and 50 Fusarium sp. nov.) obtained from sugar beet in Minnesota were characterized for pathogenicity and virulence on sugar beet in the greenhouse by a bare-root inoculation method. Among the 98 isolates tested, 80% of isolates were pathogenic: 83% of the F. oxysporum isolates, 57% of the F. graminearum isolates, and 92% of the Fusarium sp. nov. isolates. Symptoms varied from slight to moderate wilting of the foliage, interveinal chlorosis and necrosis, and vascular discoloration of the taproot without any external root symptoms. Among the pathogenic isolates, 14% were highly virulent and 12% were moderately virulent. Most of the highly virulent isolates (91%) and moderately virulent isolates (89%) were Fusarium sp. nov. All pathogenic isolates of F. graminearum and most pathogenic isolates (87%) of F. oxysporum were less virulent. In general, more-virulent isolates induced first foliar symptoms earlier compared with less-virulent isolates. This study indicates that both F. oxysporum and Fusarium sp. nov. should be used in greenhouse and be present in field studies used for screening and developing sugar beet cultivars resistant to Fusarium yellows complex for Minnesota and North Dakota.

Microdialysis combined with proteomics for protein identification in breast tumor microenvironment in vivo.

UNLABELLED: Tumor microenvironment constitutes a reservoir for proteins released from tumor cells and the host, which can contribute significantly to tumor growth and invasion. This study aims to apply a method of combining in vivo microdialysis and proteomics to identify proteins in mammary tumor interstitial fluids, a major component of tumor microenvironment. In vivo microdialysis was performed in polyomavirus middle T antigen (PyVmT) transgenic mouse mammary tumors and age-matched control wild-type mammary glands. Over four hundred proteins were identified from the microdialysis perfusates, using the Multidimensional Protein Identification Technology. Osteopontin (OPN) is one of the proteins overexpressed in breast tumor perfusates, as confirmed with immunoassays. OPN was also found to be present in tumor-associated stroma in both PyVmT and human breast tumors, using immunohistochemistry. Specifically, fibroblasts were further shown to express OPN at both mRNA and protein levels. In vitro assays showed that OPN can stimulate PyVmT breast carcinoma cell proliferation and migration. Finally, the expression of OPN was significantly higher in the peripheral blood of mice bearing breast tumors, compared to wild-type mice. Overall, microdialysis combined with proteomics is a unique technique for identifying proteins in a tumor microenvironment in vivo. Mammary fibroblasts can secrete OPN, and its overexpression in mammary tumor microenvironment may contribute significantly to mammary tumor progression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12307-010-0046-3) contains supplementary material, which is available to authorized users.

Fluctuations in number of Cercospora beticola conidia in relationship to environment and disease severity in sugar beet.

Cercospora leaf spot, caused by Cercospora beticola, is the most damaging foliar disease of sugar beet in Minnesota (MN) and North Dakota (ND). Research was conducted to characterize the temporal progression of aerial concentration of C. beticola conidia in association with the environment and disease severity in sugar beet. In 2003 and 2004, volumetric spore traps were placed within inoculated sugar beet plots to determine daily dispersal of conidia at Breckenridge, MN, and St. Thomas, ND. Plots were rated weekly for disease severity. At both locations, conidia were first collected in early July 2003 and late June in 2004. Peaks of conidia per cubic meter of air were observed with maxima in late August 2003 and in early September 2004 at both locations. Peaks of airborne conidium concentration were significantly correlated with the average temperature of daily hours when relative humidity was greater than 87%. Weekly mean hourly conidia per cubic meter of air was significantly (P < 0.01) associated with disease severity during both years and across locations. This study showed that C. beticola conidial numbers may be used to estimate potential disease severity that, with further research, could be incorporated in a disease forecasting model to rationalize Cercospora leaf spot management.

Capture and use of solar radiation, water, and nitrogen by sugar beet (Beta vulgaris L.).

Sugar beet is spring-sown for sugar production in most sugar beet-growing countries. It is grown as a vegetative crop and it accumulates yield (sugar) from very early in its growth cycle. As long as the sugar beet plants do not flower, the sugar accumulation period is indefinite and yield continues to increase. This paper reviews the success of the sugar beet crop in capturing and using solar radiation, water and mineral nitrogen resources. The prospects for improved resource capture and therefore increased sugar yield are also considered, particularly the potential to increase solar radiation interception in the future by sowing the crop in the autumn.

Multi-walled carbon nanotubes as a solid-phase extraction adsorbent for the determination of chlorophenols in environmental water samples.

Multi-walled carbon nanotubes (MWCNs) are used as adsorbent for solid-phase extraction (SPE) of several chlorophenols (CPs). CPs were adsorbed on MWCNs cartridge, then desorbed with pH 10.0 methanol, finally determined by HPLC. Under the optimized conditions, detection limits of 0.08-0.8 ng mL(-1) were obtained. The method had been applied to analyze the five CPs in tap water and river water.

The first clinical results of "wide-focus and low-pressure" ESWL.

A clinical study of the concept "wide-focus and low-pressure" extracorporal shock-wave lithotripsy (ESWL) was performed in a scientific cooperation between the Physical Institute of the University of Stuttgart and the Xixin Medical Instruments Co. Ltd. in Wuxian-Suzhou, China. In this cooperation, self-focusing electromagnetic shock-wave generator systems from the University of Stuttgart were integrated into Xixin lithotripters and installed in seven hospitals in China. A total of 297 detailed patient protocols revealed an average of 1532 shock pulses for successful treatment with no necessity for pain medication and auxiliary measures, and a stone-free rate of 86% after a follow-up of 3 months. These results are discussed in terms of the wide-focus low-pressure conditions and the mechanism of binary fragmentation by squeezing.

An arginine/glutamine difference at the juxtaposition of transmembrane domain 6 and the third extracellular loop contributes to the markedly different nucleotide selectivities of human and canine P2Y11 receptors.

The recently cloned canine P2Y11 receptor (cP2Y11) and its human homolog (hP2Y11) were stably expressed in Chinese hamster ovary cells (CHO-K1) and 1321N1 human astrocytoma cells, and their agonist selectivities and coupling efficiencies to phospholipase C and adenylyl cyclase were assessed. Adenosine triphosphate nucleotides were much more potent and efficacious at the hP2Y11 receptor than their corresponding diphosphates in promoting both inositol phosphate and cyclic AMP accumulation. In contrast, adenosine diphosphate nucleotides were considerably more potent at the cP2Y11 receptor than their corresponding triphosphate analogs. The tri- versus diphosphate specificity of the two receptors was further confirmed in studies using Ca(2+) mobilization as a measure of receptor activation under conditions that minimized nucleotide degradation. Moreover, 2-methylthioadenosine-5'-triphosphate and 2-methylthioadenosine-5'-diphosphate were 58- and 75-fold more potent than ATP and ADP, respectively, at the cP2Y11 receptor compared with only 2- to 3-fold more potent at the hP2Y11 receptor. Mutational analysis revealed that the change of Arg-265, which is located at the juxtaposition of transmembrane domain 6 and the third extracellular loop in the hP2Y11 receptor, to glutamine in the cP2Y11 receptor is at least partly responsible for the diphosphate selectivity but not the increased sensitivity to 2-thioether-substituted adenine nucleotides at the canine receptor. These results imply a key role for a positively charged arginine residue in contributing to the recognition of extracellular nucleotides by the P2Y11 receptor and perhaps other P2Y receptors.