Increased levels of CD4+ and CD8+ T cells expressing CCR1 in patients developing allograft dysfunction; a cohort study

BackgroundLeukocyte infiltration into the graft has pivotal effects on kidney transplantation outcome. The present study sought to determine whether the expression of sequential chemokine receptors on CD4⁺ and CD8⁺ T cells in human renal allograft can predict clinical episodes.MethodsBlood samples from 52 consecutive renal transplant patients were evaluated at the time of transplantation and at three times (2, 90 and 180 days) after transplantation to analyze the expression of CCR1 and CXCR3 on CD4⁺ and CD8⁺ T cells by flowcytometry. A total of 30 biopsies, including protocol biopsy (n = 24) and cause biopsy (n = 6), were investigated according to the Banff criteria.ResultsThe mean percentage of CD4⁺ and CD8⁺ T cells expressing CCR1 was significantly increased in patients with allograft dysfunction (n = 25) (p = 0.006, p = 0.004). The mean fluorescence intensity of CXCR3 on CD4⁺ and CD8⁺ T cells were found to be significantly higher in graft dysfunction than that in well-functioning grafts. (p < 0.001, p = 0.007). Receiver Operating Characteristic (ROC) Curve Analysis showed that the calculated AUC was 0.86 at the third month for CD4⁺ CCR1⁺ and CD8⁺ CCR1⁺ (p < 0.001). Multiple logistic regression analysis showed that an increase in CD4⁺ expressing CXCR3 leads to a lower risk of graft dysfunction (OR = 0.37), while an increase in CD8⁺ expressing CCR1 results in a higher risk of graft dysfunction (OR = 3.66).ConclusionDuring renal transplantation, CD4⁺ and CD8⁺ T cells expressing CCR1 were increased in patients who developed graft dysfunction. These findings may prospectively predict allograft dysfunction, and help elucidate the underlying pathogenic mechanisms.     

Effects of B cell depletion on T cell allogeneic Immune responses: A strategy to reduce allogeneic sensitization

BackgroundB cell depletion has been employed to treat antibody-mediated organ transplantation rejection, although the effects on cellular immune responses have not been extensively investigated.MethodsA model of B cell depletion used SCID/beige mice reconstituted with BALB/c splenocytes either depleted of B cells (BD) or not (BN). BD and B/N mice received C57BL/6 skin grafts and were sacrificed after 6 weeks (BD-S6 and BN-S6).ResultsRecall proliferative responses of BD-S6 splenocytes to C57BL/6 were significantly reduced compared to BN-S6, and central memory T cells' proportions (CD4⁺CD44⁺CD62L⁺ or CD8⁺CD44⁺CD62L⁺) were significantly decreased in BD-S6 spleens. Recall IFN-γ production by BD-S6 splenocytes was significantly reduced compared to BN-S6 splenocytes (p = 0.0028). Survival times of C57BL/6 heart grafts were significantly longer in SCID/beige mice reconstituted with BD-S6 splenocytes (8.5 ± 1.1 days) than for SCID/beige reconstituted with BN-S6 splenocytes (6.0 ± 1.1 days; p = 0.0006). Under cyclosporine therapy, C57BL/6 heart survival was significantly longer for SCID/beige reconstituted with BD-S6 splenocytes (17.5 ± 6.4 days) than those reconstituted with BN-S6 splenocytes (6.2 ± 1.5 days; p < 0.0001).ConclusionB cell depletion during allogeneic sensitization decreased memory T cells and recalls IFN-γ production and reduced second-set allograft rejection.     

Efeitos de dexmedetomidina em conjunto com o pré‐condicionamento isquêmico remoto em lesão de isquemia‐reperfusão renal em ratos

Background and objectivesThe aim of this study was to evaluate the effects of remote ischemic preconditioning by brief ischemia of unilateral hind limb when combined with dexmedetomidine on renal ischemia–reperfusion injury by histopathology and active caspase‐3 immunoreactivity in rats.Methods28 Wistar albino male rats were divided into 4 groups. Group I (Sham, n = 7): Laparotomy and renal pedicle dissection were performed at 65th minute of anesthesia and the rats were observed under anesthesia for 130 min. Group II (ischemia–reperfusion, n = 7): At 65th minute of anesthesia bilateral renal pedicles were clamped. After 60 min ischemia 24 h of reperfusion was performed. Group III (ischemia–reperfusion + dexmedetomidine, n = 7): At the fifth minute of reperfusion (100 μg/kg intra‐peritoneal) dexmedetomidine was administered with ischemia–reperfusion group. Reperfusion lasted 24 h. Group IV (ischemia–reperfusion + remote ischemic preconditioning + dexmedetomidine, n = 7): After laparotomy, three cycles of ischemic preconditioning (10 min ischemia and 10 min reperfusion) were applied to the left hind limb and after 5 min with group III.ResultsHistopathological injury scores and active caspase‐3 immunoreactivity were significantly lower in the Sham group compared to the other groups. Histopathological injury scores in groups III and IV were significantly lower than group II (p = 0.03 and p = 0.05). Active caspase‐3 immunoreactivity was significantly lower in the group IV than group II (p = 0.01) and there was no significant difference between group II and group III (p = 0.06).ConclusionsPharmacologic conditioning with dexmedetomidine and remote ischemic preconditioning when combined with dexmedetomidine significantly decreases renal ischemia–reperfusion injury histomorphologically. Combined use of two methods prevents apoptosis via active caspase‐3.     

The selective biomarker IL-8 identifies IFTA after kidney transplantation in blood cells

Cellular and antibody-mediated rejection processes and also interstitial fibrosis/tubular atrophy (IFTA) lead to allograft dysfunction and loss. The search for accurate, specific and non-invasive diagnostic tools is still ongoing and essential for successful treatment of renal transplanted patients. Molecular markers in blood cells and serum may serve as diagnostic tools but studies with high patient numbers and differential groups are rare. We validated the potential value of several markers on mRNA level in blood cells and serum protein level in 166 samples from kidney transplanted patients under standard immunosuppressive therapy (steroids ± mycophenolic acid ± calcineurin inhibitor) with stable graft function, urinary tract infection (UTI), IFTA, antibody-mediated rejection (ABMR), and T-cell-mediated rejection (TCMR) applying RT-PCR and ELISA. The mRNA expression of RANTES, granulysin, granzyme-B, IP-10, Mic-A and Interferon-γ in blood cells did not distinguish specifically between the different pathologies. We furthermore discovered that the mRNA expression of the chemokine IL-8 is significantly lower in samples from IFTA patients than in samples from patients with stable graft function (p < 0.001), ABMR (p < 0.001), Borderline (BL) TCMR (p < 0.001), tubulo-interstitial TCMR (p < 0.001) and vascular TCMR (p < 0.01), but not with UTI. Serum protein concentrations of granzyme-B, Interferon-γ and IL-8 did not differ between the patient groups, RANTES concentration was significantly different when comparing UTI and ABMR (p < 0.01), whereas granulysin, Mic-A and IP-10 measurement differentiated ongoing rejection or IFTA processes from stable graft function but not from each other.The measurement of IL-8 mRNA in blood cells distinguishes clearly between IFTA and other complication after kidney transplantation and could easily be used as diagnostic tool in the clinic.     

Ibrutinib suppresses alloantibody responses in a mouse model of allosensitization

BackgroundIbrutinib is a Bruton's tyrosine Kinase (BTK) antagonist that inhibits B cell receptor (BCR) signaling. Complete BTK deficiency is associated with absence of B-cells. Ibrutinb is currently approved by FDA for treatment of B-cell malignancies, including Waldenström macroglobulinaemia. We recently carried out studies to determine if ibrutinib could modify alloantibody responses.Materials and methodsA mouse model of allogenic sensitization using a C57BL/6 mouse as the recipient of a skin allograft from an HLA-A2 transgenic mouse was utilized to examine the effects of ibrutinib on alloantibody responses and B cell effector functions. Donor-specific antibody (DSA) levels were measured in a flow-cytometric antibody binding assay. Splenic T and B cell subsets and plasma cells were analyzed in flow cytometry.ResultsControl mice developed peak levels of DSA IgM at day 14 PTx while the ibrutinib treated mice had significantly lower levels of DSA IgM (p = 0.0047). Control mice developed HLA.A2-specific IgG antibodies at day 14 (230 ± 60 MFI) and reached peak levels at day 21 (426 ± 61 MFI). In contrast, mice in the treatment group had low levels of HLA.A2-specific IgG at day 14 (109 ± 59 MFI, p = 0.004) and day 21 (241 ± 86 MFI, p = 0.003). FACS analysis found a reduction of B220⁺ or CD19⁺ B cell population (p < 0.05). In addition, ibrutinib attenuated recall DSA IgG responses to re-sensitization (p < 0.05) and reduced CD38⁺ CD138⁺ plasma cells (p < 0.05) in the spleens.ConclusionsIbrutinib is effective in suppressing alloantibody responses through blocking BTK-mediated BCR signaling, leading to reduction of B cells and short-lived plasma cells in the spleens. Use of ibrutinib may provide benefits to HLA-sensitized transplant patients for alloantibody suppression.     

Absence of CD4CD25 regulatory T cell expansion in renal transplanted patients treated in vivo with Belatacept mediated CD28–CD80/86 blockade

AimsBelatacept is a new recombinant molecule (CTLA4-Ig) that interferes with the second activation signal of T lymphocytes. CTLA4-Ig induced T cell allograft tolerance in rodents but not in primates. We examined the changes in peripheral lymphocyte subsets, including regulatory T cells, in renal transplant patients treated with Belatacept.MethodsA cross-sectional immunological study was carried out 6 months after transplantation in 28 patients enrolled in the Belatacept phase II study. Eighteen patients received Belatacept, mycophenolate mofetil and steroids (Belatacept group), while the control group of 10 patients received cyclosporine, mycophenolate mofetil and steroids (CsA group). Lymphocyte subsets were examined by flow cytometry. Foxp3 mRNA expression was measured by quantitative PCR.ResultsThe number of T lymphocytes and the percentage of CD3+ T cells were similar in both groups. However, the percentage of CD3+ CD4+ T cells was lower in the Belatacept group than in the control CsA group (B = 42.5% ± 13.7 vs CsA = 52.9% ± 9, p < 0.005), and the percentage of CD3+ CD8+ cells was higher in the Belatacept group than in the control (B = 32.9% ± 6.7 vs CsA = 19.5% ± 8.2, p < 0.0002). The percentage of CD19+ cells was similar in both groups. Among CD56+cells, only the percentage of CD16+ cells was significantly higher in the Belatacept group than in the control (B = 82% ± 12 vs CsA = 59.7% ± 25, p = 0.01). Among CD4 and CD8 T cells the percentage of activated lymphocytes expressing CTLA4, HLA-DR or CD40L was similar in both groups. The percentage of CD4+CD25+ T cells was higher in the CsA group. The percentage of regulatory CD4+CD25+ cells with bright CD25 staining was similar in both groups (B = 3.6 ± 2.3% vs CsA = 4.7 ± 1.9%, ns) as was the expression of FoxP3.ConclusionOur results indicated that Belatacept did not induce regulatory T cell expansion in vivo. We suggest that Belatacept treatment should be maintained after transplantation to allow graft acceptance.     

IFNy + and IFNy − Treg subsets with stable and unstable Foxp3 expression in kidney transplant recipients with good long-term graft function

BackgroundTreg are a heterogenous cell population. In the present study we attempted to identify Treg subsets that might contribute to stable and good long-term graft function.MethodLymphocyte and Treg subsets were studied in 136 kidney transplant recipients with good long-term graft function and in 52 healthy control individuals using eight-color-fluorescence flow cytometry. Foxp3 TSDR methylation status was investigated in enriched IFNy + and IFNy − Treg preparations using high resolution melt analysis.ResultsCompared with healthy controls, patients showed strong associations of IFNy secreting Helios + and Helios − Treg with Treg that co-expressed perforin and/or CTLA4 (CD152; p < 0.01). Moreover they showed associations of IFNy − Helios + Treg with Treg that produced TGFβ and/or perforin and of IFNy − Helios − Treg with TGFβ production (all p < 0.01). Only in patients, but not in healthy controls, were IFNy − Helios + and Helios − Treg associated with higher CD45 +, CD3 +, (CD4 +), CD19 + lymphocyte counts (p < 0.001). In addition IFNy − Helios + Treg were associated with CD16 + 56 + lymphocytes (p < 0.001). Enriched IFNy − Treg from female but not male patients showed an association of Foxp3 methylation with higher total Treg and higher Helios + IFNy −, CXCR3 + Lselectin + (CD183 + CD62L +), CXCR3 − Lselectin + and CD28 + HLADR + Treg subsets (p < 0.01). Enriched IFNy + Treg from male patients showed an association of demethylated Foxp3 with total Treg and IL10 − TFGβ + Treg counts, and in enriched IFNy − Treg an association of methylated Foxp3 with APO1/FasR + FasL + (CD95 + CD178 +) Treg (p < 0.01).ConclusionsKidney recipients with good long-term graft function possess IFNy + and IFNy − Treg with stable and unstable Foxp3 expression in the blood. They co-express CD28, HLADR, CTLA4, CXCR3, Lselectin, TGFβ, perforin and FasL and might contribute to the establishment and maintenance of good long-term graft function.     

Adjuvant alpha adrenergic blockers: Are they equally efficient for renal and upper ureteral calculi disintegrated by shock wave lithotripsy?

ObjectivesTo evaluate the effects of tamsulosin on stone clearance and analgesic requirements after shock wave lithotripsy (SWL) for solitary renal and upper ureteral calculi.Patients and methodsA prospective randomized placebo controlled study was carried out on 126 patients who underwent SWL for solitary radio-opaque renal or upper ureteral calculi ≤20 mm. Patients were randomized into two groups receiving either 0.4 mg of tamsulosin (GT) or placebo (GP). SWL was performed 3-weekly until patients became stone-free or for a maximum of 3 months. Analgesics were used on demand and pain was evaluated by a visual pain scale.ResultsRenal stones represented 55.6% and 66.7% for GT and GP, respectively (p = 0.27). Mean renal and ureteral stone size were (12.3 ± 1.8 mm vs. 11.5 ± 2.3 mm, p = 0.14) and (9.7 ± 2.6 mm vs. 8.6 ± 1.7 mm, p = 0.1) for the GT and GP, respectively. GT required fewer SWL sessions for ureteral (1.2 vs. 1.6, p = 0.02) and renal stones (1.8 vs. 2.3, p = 0.08). Stone-free rate (SFR) was higher in GT for upper ureteral stones (96.4% vs. 66.7%, p = 0.01) and renal pelvis stones at a cutoff size >10 mm (p = 0.01). The mean time of stone clearance was significantly lower in GT (4.2 ± 1.9 weeks vs. 7.5 ± 2.3 weeks, p = 0.001) for ureteral stones. Attacks of renal colic were more frequent in GP (82.5% vs. 44.4%, p = 0.04) with increased demand for analgesia (p = 0.04). Steinstrasse was recorded in 3 and 7 patients of the GT and GP, respectively (p = 0.32).ConclusionTamsulosin facilitates clearance of upper ureteral stone fragments after SWL and decreases the analgesic requirements. These effects were not similarly evident for renal stones.     

Donor age negatively affects the immunoregulatory properties of both adipose and bone marrow derived mesenchymal stem cells

PurposeAge negatively impacts the biologic features of mesenchymal stem cells (MSCs), including decreased expansion kinetics and differentiation potential. Clinically, donor-age may be within a wide spectrum; therefore, investigation of the role of donor's age on immunoregulatory potential is of critical importance to translate stem cell therapies from bench to bedside.MethodsAdipose and bone marrow derived MSCs (ASCs and BMSCs) were isolated in parallel from Lewis and Brown Norway rats of young (less than 4-week old) and senior groups (older than 15-month). The presentation of cells and time required for growth to 90% confluence was recorded. FACS sorting based on the expression of CD90 and CD29 double positive and CD45 CD11 double negative quantified the proportions of MSCs. After expansion, ASCs and BMSCs from different age groups were co-cultured in mixed lymphocyte reaction (MLR; Lewis vs. Brown Norway) assays. The suppression of CD3⁺CD4⁺ and CD3⁺CD8⁺ T cell populations by different sources of MSCs were compared.ResultsThe kinetics of cell growth was slower in old animals (17.3 ± 2 days) compared with young animals (8.8 ± 3 days), and cell morphology was irregular and enlarged in the senior groups. The yield of MSCs by FACS sorting was significantly higher in young groups compared to senior groups (p < 0.02). With regard to immunoregulatory potential, senior ASCs failed to induce any CD3⁺CD4⁺ T cell suppression (p > 0.05). In addition, young BMSCs-induced suppression was more prominent than seniors (p < 0.05).ConclusionsDonor age should be taken into consideration when using recipient MSC of either bone marrow or adipose origin in clinical applications.     

The effect on improvement of recovery and pain scores of paravertebral block immediately before breast surgery

ObjectivesParavertebral block (PVB) has the potential to reduce postoperative pain after breast surgery. The aim of the study was to investigate whether PVB performed immediately before surgery could affect the postoperative morbidities in terms of pain and emesis, and improve the quality of recovery (QoR) in patients after surgery for breast cancer.MethodsPostoperative data were collected prospectively from two groups of patients undergoing unilateral breast surgery during the study period of 1 month. Forty consecutive patients received either solely general anesthesia (GA group, n = 25) or GA plus ultrasound-guided PVB (GA + PVB group, n = 15) for the surgery. Pain scores and areal distribution of pain were compared between the two groups 1 hour and 6 hours postoperatively and on the midmorning of postoperative Day 1 (POD1). The QoR scores were compared between the two groups 6 hours postoperatively and on the midmorning of POD1. Incidence of postoperative nausea and vomiting and doses of analgesics and narcotics given were also compared.ResultsPain scores at rest were significantly lower in the GA + PVB group at all designated time points [1 hour (p < 0.0001), 6 hours (p < 0.0001), and on midmorning of POD1 (p = 0.041)]. Pain scores with movements was also significantly lower at all time points in the GA + PVB group (1 hour, p < 0.0001; 6 hours, p < 0.0001; midmorning of POD1, p = 0.0012). Areal distribution of pain at rest and with movement was wider in the GA group 1 hour and 6 hours postoperately but was identical to that of GA+ PVB group on the mid-morning of POD1 [1 hour postoperatively at rest (p < 0.0001), with movement (p < 0.0001); 6 hours postoperatively at rest (p = 0.0018), with movement (p = 0.0048)]. The QoR scores were significantly higher in the GA + PVB group at 6 hours (p < 0.0001) and on midmorning of POD1 (p = 0.0079). The incidences of postoperative nausea and vomiting were significantly lower in the GA + PVB group (p = 0.0004). Doses of postoperative analgesics and narcotics were significantly less in the GA + PVB group (p < 0.0001 and p = 0.001, respectively). Time to first request for analgesics was significantly longer in the GA + PVB group (p = 0.0002).ConclusionsPVB given before surgery in combination with GA could provide better postoperative analgesia and better QoR than did GA alone in patients undergoing surgery for unilateral breast cancer.     

High proportion of CD95+ and CD38+ in cultured CD8+ T cells predicts acute rejection and infection,respectively, in kidney recipients

The aim of this study was to find noninvasive T-cell markers able to predict rejection or infection risk after kidney transplantation.We prospectively examined T-lymphocyte subsets after cell culture stimulation (according to CD38, CD69, CD95, CD40L, and CD25 expression) in 79 first graft recipients from four centers, before and after transplantation. Patients were followed up for one year.Patients who rejected within month-1 (n = 10) showed high pre-transplantation and week-1 post-transplantation percentages of CD95⁺, in CD4⁺ and CD8⁺ T-cells (P < 0.001 for all comparisons). These biomarkers conferred independent risk for early rejection (HR:5.05, P = 0.061 and HR:75.31, P = 0.004; respectively). The cut-off values were able to accurately discriminate between rejectors and non-rejectors and Kaplan–Meier curves showed significantly different free-of-rejection time rates (P < 0.005). Patients who rejected after the month-1 (n = 4) had a higher percentage of post-transplantation CD69⁺ in CD8⁺ T-cells than non-rejectors (P = 0.002). Finally, patients with infection (n = 41) previously showed higher percentage of CD38⁺ in CD8⁺ T-cells at all post-transplantation times evaluated, being this increase more marked in viral infections. A cut-off of 59% CD38⁺ in CD8⁺ T-cells at week-1, week-2 and month-2 reached 100% sensitivity for the detection of subsequent viral infections.In conclusion, predictive biomarkers of rejection and infection risk after transplantation were detected that could be useful for the personalized care of kidney recipients.     

NKT cells are important mediators of hepatic ischemia-reperfusion injury

IntroductionIRI results from the interruption then reinstatement of an organ's blood supply, and this poses a significant problem in liver transplantation and resectional surgery.In this paper, we explore the role T cells play in the pathogenesis of this injury.Materials & methodsWe used an in vivo murine model of warm partial hepatic IRI, genetically-modified mice, in vivo antibody depletion, adoptive cell transfer and flow cytometry to determine which lymphocyte subsets contribute to pathology. Injury was assessed by measuring serum alanine aminotransfersase (ALT) and by histological examination of liver tissue sections.ResultsThe absence of T cells (CD3εKO) is associated with significant protection from injury (p = 0.010). Through a strategy of antibody depletion it appears that NKT cells (p = 0.0025), rather than conventional T (CD4 + or CD8 +) (p = 0.11) cells that are the key mediators of injury.DiscussionOur results indicate that tissue-resident NKT cells, but not other lymphocyte populations are responsible for the injury in hepatic IRI. Targeting the activation of NKT cells and/or their effector apparatus would be a novel approach in protecting the liver during transplantation and resection surgery; this may allow us to expand our current criteria for surgery.     

Balloon Angioplasty as the Primary Treatment for Failing Infra-inguinal Vein Grafts

BackgroundWe sought to evaluate the role of balloon angioplasty as the primary modality in the management of vein graft stenoses.MethodsPatients who underwent infrainguinal vein graft bypass from January 2002 to December 2007 were enrolled into a surveillance program. Grafts which developed critical stenoses were identified and underwent urgent angiography with a view to angioplasty of the stenotic lesion. Lesions which were deemed unsuitable for angioplasty underwent urgent surgical repair.ResultsFour hundred and eleven grafts were followed up for a median of 19 months (range: 2–61). Ninety-six grafts (22.6%) developed critical stenosis. Twelve grafts occluded prior to repair and one was not intervened upon electively. Eight grafts underwent primary surgical repair. Seventy-six grafts underwent 99 endovascular procedures. Technical success was achieved in 60 grafts (78.9%). Of the grafts in which technical success had not been achieved, eight underwent repeat angioplasty and three were surgically repaired. Twenty-four grafts underwent repeat angioplasty for re-stenosis with a technical success rate of 71%. No difference was observed in graft patency (P = 0.08) or amputation rates (P = 0.32) between the grafts requiring intervention to maintain patency, and grafts which did not. Smoking [OR: 2.61 (95% CI: 1.51–4.53), (P = 0.006)], diabetes [OR: 2.55 (95% CI: 1.49–4.35), (P = 0.006)], renal failure [OR: 1.89 (95% CI: 1.19–3.38), (P = 0.040)] and recurrent stenosis [OR: 3.22 (95% CI: 1.63–4.69), (P < 0.001)] were risk factors for graft occlusion.ConclusionsBalloon angioplasty of failing infrainguinal vein bypass grafts is safe and can be performed with an acceptable medium term patency rate, albeit with a significant risk of re-stenosis which can be successfully treated in most patients using repeat endovascular intervention.     

Characterisation of Interleukin-8 and Monocyte Chemoattractant Protein-1 Expression within the Abdominal Aortic Aneurysm and their Association with Mural Inflammation

ObjectivesAbdominal aortic aneurysms (AAAs) are characterised by chronic transmural inflammation. This study investigated the expression of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) within the AAA, and their relationship with mural inflammation.MethodsBiopsies were obtained from 25 AAAs, 15 abdominal aortas, and 10 atherosclerotic thoracic aortas. IL-8 and MCP-1 expression was measured in homogenised specimens by ELISA. Infiltrate composition and localised expression of IL-8 and MCP-1 were determined through immunohistochemistry.ResultsELISA analysis demonstrated that IL-8 and MCP-1 were raised in the AAA compared to the controls [(IL-8, AAA vs. abdominal aorta: >28-fold, P < .001; AAA vs. thoracic aorta: >28-fold, P < .001) (MCP-1, AAA vs. abdominal aorta: 9-fold, P < .001; AAA vs. thoracic aorta: 19-fold, P < .001)]. Immunohistochemistry revealed that IL-8 was localised to the inflammatory infiltrate, which consisted predominantly of CD3⁺ T- and CD20⁺ B-lymphocytes. MCP-1 was predominantly expressed by CD68⁺ macrophages. Increasing IL-8 expression was associated with an increase in mural inflammation, and an increase in CD3⁺ T-lymphocytes of CD4⁺ phenotype within the infiltrate population.ConclusionPathways involving IL-8 and MCP-1 may be involved in AAA pathogenesis. IL-8 may be directly involved in the chemotaxis of T_H-lymphocytes into the AAA wall.     

Low osteocalcin/collagen type I bone gene expression ratio is associated with hip fragility fractures

IntroductionOsteocalcin (OC) is the most abundant non-collagenous bone protein and is determinant for bone mineralization.We aimed to compare OC bone expression and serum factors related to its carboxylation in hip fragility fracture and osteoarthritis patients. We also aimed to identify which of these factors were associated with worse mechanical behavior and with the hip fracture event.MethodsIn this case-control study, fragility fracture patients submitted to hip replacement surgery were evaluated and compared to a group of osteoarthritis patients submitted to the same procedure. Fasting blood samples were collected to assess apolipoproteinE (apoE) levels, total OC and undercarboxylated osteocalcin (ucOC), vitamin K, LDL cholesterol, triglycerides and bone turnover markers. The frequency of the apoε4 isoform was determined.Femoral epiphyses were collected and trabecular bone cylinders drilled in order to perform compression mechanical tests. Gene expression of bone matrix components was assessed by quantitative RT-PCR analysis.Results64 patients, 25 submitted to hip replacement surgery due to fragility fracture and 39 due to osteoarthritis, were evaluated. Bone OC/collagen expression (OC/COL1A1) ratio was significantly lower in hip fracture compared to osteoarthritis patients (p < 0.017) adjusted for age, gender and body mass index. Moreover, OC/COL1A1 expression ratio was associated with the hip fracture event (OR ~ 0; p = 0.003) independently of the group assigned, or the clinical characteristics. Apoε4 isoform was more frequent in the hip fracture group (p = 0.029). ucOC levels were higher in the fracture group although not significantly (p = 0.058). No differences were found regarding total OC (p = 0.602), apoE (p = 0.467) and Vitamin K (p = 0.371).In hip fracture patients, multivariate analysis, adjusted for clinical characteristics, serum factors related to OC metabolism and gene expression of bone matrix proteins showed that low OC/COL1A1 expression ratio was significantly associated with worse trabecular strength (β = 0.607; p = 0.013) and stiffness (β = 0.693; p = 0.003). No association was found between ucOC and bone mechanics. Moreover, in osteoarthritis patients, the multivariate analysis revealed that serum total OC was negatively associated with strength (β = ? 0.411; p = 0.030) and stiffness (β = ? 0.487; p = 0.009).ConclusionWe demonstrated that low bone OC/COL1A1 expression ratio was an independent predictor of worse trabecular mechanical behavior and of the hip fracture event. These findings suggest that in hip fracture patients the imbalance of bone OC/COL1A1 expression ratio reflects disturbances in osteoblast activity leading to bone fragility.     

Low osteocalcin/collagen type I bone gene expression ratio is associated with hip fragility fractures

IntroductionOsteocalcin (OC) is the most abundant non-collagenous bone protein and is determinant for bone mineralization.We aimed to compare OC bone expression and serum factors related to its carboxylation in hip fragility fracture and osteoarthritis patients. We also aimed to identify which of these factors were associated with worse mechanical behavior and with the hip fracture event.MethodsIn this case-control study, fragility fracture patients submitted to hip replacement surgery were evaluated and compared to a group of osteoarthritis patients submitted to the same procedure. Fasting blood samples were collected to assess apolipoproteinE (apoE) levels, total OC and undercarboxylated osteocalcin (ucOC), vitamin K, LDL cholesterol, triglycerides and bone turnover markers. The frequency of the apoε4 isoform was determined.Femoral epiphyses were collected and trabecular bone cylinders drilled in order to perform compression mechanical tests. Gene expression of bone matrix components was assessed by quantitative RT-PCR analysis.Results64 patients, 25 submitted to hip replacement surgery due to fragility fracture and 39 due to osteoarthritis, were evaluated. Bone OC/collagen expression (OC/COL1A1) ratio was significantly lower in hip fracture compared to osteoarthritis patients (p < 0.017) adjusted for age, gender and body mass index. Moreover, OC/COL1A1 expression ratio was associated with the hip fracture event (OR ~ 0; p = 0.003) independently of the group assigned, or the clinical characteristics. Apoε4 isoform was more frequent in the hip fracture group (p = 0.029). ucOC levels were higher in the fracture group although not significantly (p = 0.058). No differences were found regarding total OC (p = 0.602), apoE (p = 0.467) and Vitamin K (p = 0.371).In hip fracture patients, multivariate analysis, adjusted for clinical characteristics, serum factors related to OC metabolism and gene expression of bone matrix proteins showed that low OC/COL1A1 expression ratio was significantly associated with worse trabecular strength (β = 0.607; p = 0.013) and stiffness (β = 0.693; p = 0.003). No association was found between ucOC and bone mechanics. Moreover, in osteoarthritis patients, the multivariate analysis revealed that serum total OC was negatively associated with strength (β = − 0.411; p = 0.030) and stiffness (β = − 0.487; p = 0.009).ConclusionWe demonstrated that low bone OC/COL1A1 expression ratio was an independent predictor of worse trabecular mechanical behavior and of the hip fracture event. These findings suggest that in hip fracture patients the imbalance of bone OC/COL1A1 expression ratio reflects disturbances in osteoblast activity leading to bone fragility.     

The effect of aortic stenosis on elderly hip fracture outcomes: A case control study

IntroductionAortic stenosis (AS) is an established predictor of perioperative complications following both cardiac and non-cardiac surgery. The purpose of this study was to evaluate the risk of mortality and perioperative complications among surgically treated hip fractures in elderly patients with moderate or severe AS compared to those without AS (negative controls).Materials and methodsA retrospective case-controlled review (1:2) of elderly (≥65 years) surgically treated hip fractures from 2011 to 2015 with moderate/severe AS (according to American Heart Association criteria) was conducted. Postoperative complication rates, 30 days and 1 year mortality were reviewed.ResultsModerate/severe AS was identified in 65 hip fracture cases and compared to 129 negative controls. AS cases were significantly older with higher rates of coronary artery disease and atrial fibrillation (p < 0.05). Rates of any 30-day perioperative complication (74% vs. 37%, p < 0.001) and severe non-cardiac 30-day perioperative complication (52% vs. 26%, p = 0.002) were significantly higher among AS cases compared to controls. Kaplan Meier estimates of 30-day mortality (14.7% vs. 4.2%, p < 0.001) and 1-year mortality (46.8% vs. 14.1%, p < 0.001) were significantly higher in AS cases compared to controls. Multivariate analysis of severe 30-day postoperative complications identified moderate/severe AS (OR 4.02, p = 0.001), pulmonary disease (OR 7.36, p = 0.002) and renal disease (OR 3.27, p = 0.04) as independent predictors. Moderate/severe AS (OR 3.38, p = 0.03), atrial fibrillation (OR 3.73, p = 0.03) and renal disease (OR 4.44, p = 0.02) were independent predictors of 30-day mortality. Moderate/severe AS (OR 5.79, p < 0.001) and renal disease (OR 3.39, p = 0.02) were independent predictors of 1-year mortality.ConclusionAortic stenosis is associated with a significantly increased risk of perioperative complications, 30-day mortality and 1-year mortality in elderly patients undergoing surgical treatment of hip fractures.     

Short-Term Results of A Randomized Trial Comparing Remote Endarterectomy and Supragenicular Bypass Surgery for Long Occlusions of the Superficial Femoral Artery [The REVAS Trial]

ObjectiveTechniques for surgical repair of Trans-Atlantic Inter-Society Consensus (TASC) C and D lesions of the superficial femoral artery (SFA) are supragenicular bypass grafting or the less invasive remote endarterectomy (RSFAE). This trial compares the patency rates of both techniques.DesignRandomized, multicenter trial.Materials and methods116 patients were randomized to RSFAE (n = 61) and supragenicular bypass surgery (n = 55). Indications for surgery were claudication (n = 77), rest pain (n = 21), or tissue loss (n = 18).ResultsMedian hospital stay was 4 days in the RSFAE group compared with 6 days in the bypass group (p = 0.004). Primary patency after 1-year follow-up was 61% for RSFAE and 73% for bypass (p = 0.094). Secondary patency was 79% for both groups. Subdividing between venous (n = 25) and prosthetic grafts (n = 30) shows a primary patency of 89% and 63% respectively at 1-year follow-up (p = 0.086).ConclusionRSFAE is a minimally invasive adjunct in the treatment of TASC C and D lesions of the SFA, with shorter admittance and a comparable secondary patency rate to bypass. The venous bypass is superior to both RSFAE and PTFE bypass surgery, but only 45% of patients had a sufficient saphenous vein available.This study is registered with ClinicalTrials.gov, number NCT00566436.     

Prostate-only Versus Whole-pelvis Radiation with or Without a Brachytherapy Boost for Gleason Grade Group 5 Prostate Cancer: A Retrospective Analysis

BackgroundThe role of elective whole-pelvis radiotherapy (WPRT) remains controversial. Few studies have investigated it in Gleason grade group (GG) 5 prostate cancer (PCa), known to have a high risk of nodal metastases.ObjectiveTo assess the impact of WPRT on patients with GG 5 PCa treated with external-beam radiotherapy (EBRT) or EBRT with a brachytherapy boost (EBRT + BT).Design, setting, and participantsWe identified 1170 patients with biopsy-proven GG 5 PCa from 11 centers in the United States and one in Norway treated between 2000 and 2013 (734 with EBRT and 436 with EBRT + BT).Outcome measurements and statistical analysisBiochemical recurrence-free survival (bRFS), distant metastasis-free survival (DMFS), and prostate cancer-specific survival (PCSS) were compared using Cox proportional hazards models with propensity score adjustment.Results and limitationsA total of 299 EBRT patients (41%) and 320 EBRT + BT patients (73%) received WPRT. The adjusted 5-yr bRFS rates with WPRT in the EBRT and EBRT + BT groups were 66% and 88%, respectively. Without WPRT, these rates for the EBRT and EBRT + BT groups were 58% and 78%, respectively. The median follow-up was 5.6 yr. WPRT was associated with improved bRFS among patients treated with EBRT + BT (hazard ratio [HR] 0.5, 95% confidence interval [CI] 0.2–0.9, p = 0.02), but no evidence for improvement was found in those treated with EBRT (HR 0.8, 95% CI 0.6–1.2, p = 0.4). WPRT was not significantly associated with improved DMFS or PCSS in the EBRT group (HR 1.1, 95% CI 0.7–1.7, p = 0.8 for DMFS and HR 0.7, 95% CI 0.4–1.1, p = 0.1 for PCSS), or in the EBRT + BT group (HR 0.6, 95% CI 0.3–1.4, p = 0.2 for DMFS and HR 0.5 95% CI 0.2–1.2, p = 0.1 for PCSS).ConclusionsWPRT was not associated with improved PCSS or DMFS in patients with GG 5 PCa who received either EBRT or EBRT + BT. However, WPRT was associated with a significant improvement in bRFS among patients receiving EBRT + BT. Strategies to optimize WPRT, potentially with the use of advanced imaging techniques to identify occult nodal disease, are warranted.Patient summaryWhen men with a high Gleason grade prostate cancer receive radiation with external radiation and brachytherapy, the addition of radiation to the pelvis results in a longer duration of prostate-specific antigen control. However, we did not find a difference in their survival from prostate cancer or in their survival without metastatic disease. We also did not find a benefit for radiation to the pelvis in men who received radiation without brachytherapy.     

Low utility of serum 25–hydroxyvitamin D3 and 1, 25-dihydroxyvitamin D3 in predicting peripheral Treg and Th17 cell counts in ESRD and renal transplant patients

BackgroundVitamin D has shown an immune-modulatory effect in different studies. Vitamin D stimulates Tregs and inhibits Th17 cells. The immune-modulatory role of vitamin D in chronic kidney disease (CKD) and renal transplant patients is unclear. We measured whether different serum levels of vitamin D were associated with an increased or decreased presence of lymphocyte subsets including Treg and Th17 cells in end-stage renal disease (ESRD) and renal transplant recipients.MethodsEighty-seven renal transplant recipients and 53 end-stage renal disease (ESRD) patients were enrolled in this study. The absolute counts of CD4 + and CD8 + T, CD16 + CD56 + NK, CD19 + B, CD4 + CD25 + CD127- Foxp3 + (Tregs), Helios + Tregs, CD38 + Tregs, and CD4 + CD17 + (Th17) cells were analyzed in peripheral blood in both patient groups. In addition, serum 25 (OH) D₃, 1, 25 (OH)₂ D₃, IL-6, IL-17, IL-23, and TGF-β₁ were measured. The association between lymphocyte subset counts and 1, 25 (OH)₂ D₃ or 25 (OH) D₃ was studied, as was the association between serum IL-6, IL-17, IL-23, or TGF-β₁ and 1,25 (OH)₂ D₃ or 25 (OH) D₃.ResultsSerum 25 (OH) D₃ and 1,25 (OH)₂ D₃ levels were not independently associated with peripheral CD4 + T, CD19 + B, CD16 + CD56 + NK, Treg, or Th17 cell counts. In contrast to serum 25 (OH) D₃, serum1, 25 (OH)₂ D₃ was positively associated with CD8 + T cells counts in renal transplant recipients.ConclusionOur findings indicate low utility of serum 25 (OH) D₃ and 1, 25 (OH)₂ D₃ levels in predicting a change in lymphocyte subset counts in ESRD and renal transplant patients.