A novel pattern of pp65-positive cytomegalic endothelial cells circulating in peripheral blood from a renal transplant recipient

The present study reports a novel pattern of cytomegalic endothelial cells (CEC) in peripheral blood from a female renal transplant recipient infected with human cytomegalovirus (HCMV), which has not been reported previously. Localization of specific early antigen of HCMV, pp65 antigen, was examined by immunohistochemistry. Staining of an endothelial cell marker (CD34) was used to characterize endothelial cells. It is demonstrated that many leukocytes surrounded and adhered to a protein-like material, in which pp65-positive CEC were detected. The composition and function of this protein-like material are yet unknown. The patient lacked clinical symptoms of HCMV disease. Furthermore, similar localization patterns were found in other renal transplant recipients suffering from HCMV infections as determined by real-time PCR to detect HCMV DNA in blood. These patients showed no or only minor clinical symptoms of HCMV infection. It is suggested that these novel localization patterns of CEC may play a role in the host defense in patients infected with HCMV, but the exact relation between HCMV infection and CEC formation needs further investigation.     

Circulating endothelial cells in Kawasaki disease

Recent reports have demonstrated that circulating endothelial cells (CECs) are observed in several diseases with vascular injury. Because Kawasaki disease (KD) is one type of systemic vasculitis, we hypothesized that an increased number of CECs may be associated with the appearance of complicated coronary artery lesions (CAL). In the present study we investigated the enumeration and origin of CECs in 20 patients with KD, using an immunohistochemical method with monoclonal antibodies: clone P1H12 against ECs and clone AC133 against endothelial progenitor cells (EPCs), which were derived from the bone marrow. The mean number of CECs increased significantly (P < 0.05) from the acute through the subacute phases of KD compared with both the convalescent phase of KD and healthy children. The mean number of CECs was significantly (P < 0.05) higher in six KD patients with CAL than in 14 KD patients without CAL. The population of EPCs in the total CECs in KD was 4.4 +/- 1.2% (range 0-18%). The number of EPCs during the subacute phase was also significantly higher (P < 0.05) in KD patients with CAL than in those without CAL. Our findings indicate that the number of CECs increase in KD vasculitis and suggest that the increased numbers of CECs and EPCs may reflect the EC damage of this disease.     

Circulating cytomegalic endothelial cells are associated with high human cytomegalovirus (HCMV) load in AIDS patients with late-stage disseminated HCMV disease

The prevalence of circulating cytomegalic endothelial cells, detected currently by the pp65-antigenemia assay and described previously in blood of transplanted and AIDS patients with disseminated human cytomegalovirus (HCMV) infection, was found to be 2.9% in the AIDS population and 6.5% in the fraction of the AIDS population with HCMV in blood. Cytomegalic endothelial cells increased to 39.7% and 48.4%, respectively, in AIDS patients with very high levels of antigenemia and viremia, while an end organ disease reached an incidence of 76.4%. Positive and negative predictive values of cytomegalic endothelial cell detection for diagnosis of HCMV end organ disease were 73.1% and 21.4% with antigenemia levels >1,000, respectively. On the other hand, in a selected group of 38 cytomegalic endothelial cell-positive AIDS patients with <50 CD4+ T cells/μl and late-stage HCMV disease, who were followed-up for variable periods of time, the prevalence of high level antigenemia was 95.3%, that of viremia 86.0% and that of L-DNAemia 92.7%, while the incidence of HCMV end organ disease was 84.2%. In this population, it was shown that cytomegalic endothelial cell presence was associated with lack of (56.0% of episodes) or insufficient (4.0%) anti-HCMV treatment or emergence of HCMV drug-resistant strains (17.3%) or short-term response to antiviral treatment (22.7%); was determined in the same patient by different conditions during follow-up. Longitudinal observations indicated that cytomegalic endothelial cells were detected often in blood at least 3 months later than end organ disease suggesting that the duration of end organ disease was a cofactor associated with the appearance of cytomegalic endothelial cells. J. Med. Virol. 55:64–74, 1998. © 1998 Wiley-Liss, Inc.     

Circulating endothelial cells in patients with heart failure and left ventricular dysfunction

Introduction and Aims: Acute and chronic heart failure may manifest different degrees of endothelial damage and angiogenesis. Circulating endothelial cells (CEC) have been identified as marker of vascular damage. The aim of our study was to evaluate the evolution of the CEC at different stages of patients with heart failure. We also investigated a potential correlation between CEC and markers of vascular damage and angiogenesis. Methods: We studied 32 heart failure patients at hospital admission (acute phase) and at revision after 3 months (stable phase) and 32 controls. Circulating markers of endothelial damage (CEC; von Willebrand factor, vWF and soluble E-selectin, sEsel) and angiogenesis (vascular endothelial growth factor, VEGF and thrombospondin-1) were quantified. Results: Levels of CEC, vWF, sEsel and VEGF are significantly higher in heart failure patients than in controls. Levels of CEC (36.9 ± 15.3 vs. 21.5 ± 10.0 cells/ml; p< 0.001), vWF (325 ± 101 vs. 231 ± 82%; p< 0.001) and VEGF (26.3 ± 15.2 vs. 21.9 ± 11.9 ng/ml; p< 0.001) are significantly higher in the acute phase than in the stable phase of heart failure. CEC levels correlate with vWF and VEGF. Results show than 100% of patients in acute phase and 37.5% in stable phase have levels of CEC higher than the 99th percentile of the distribution of controls (16 cells/ml). Therefore, increases in CEC represent a relative risk of 9.5 for heart failure patients suffering from acute phase. Conclusions: CEC, in addition to being elevated in heart failure, correlate with vWF levels, providing further support for CEC as markers of endothelial damage. Levels of CEC are associated with the acute phase of heart failure and could be used as a marker of the worsening in heart failure.     

Circulating endothelial progenitor cells during normal pregnancy and pre-eclampsia

PROBLEM: Endothelial progenitor cell (EPC), which mediates neovascularization of uterine endometrium may be involved in the neovascularization in the utero-placental circulation. We evaluated whether EPC proliferation in pre-eclampsia (PE) differed from that in normal pregnancy. METHOD OF STUDY: EPC number in peripheral blood (20 non-pregnancy, 36 normal pregnancy, 10 PE) was measured using flow cytometry. Peripheral blood mononuclear cell was cultured for 7 days and EPC proliferation was assessed based on detection of the uptake of acetylated low-density lipoprotein and lectin. Furthermore, the proliferative activity induced by angiotensin II (Ang II) and tumor necrosis factor-alpha (TNF-alpha) was measured by BrdU assay. RESULTS: EPC number in peripheral blood did not differ significantly between PE and normal pregnancy; however, EPC proliferation was significantly increased in PE. Furthermore, Ang II and TNF-alpha induced the proliferation of EPC derived from patients with PE. CONCLUSIONS: In PE, some factors including Ang II and TNF-alpha stimulated EPC proliferation; however, the impairment of EPC mobilization into systemic circulation by serum factors may contribute to insufficient regeneration of EC in disturbed utero-placental circulation of PE.     

Enhanced endothelial cytopathogenicity induced by a cytomegalovirus strain propagated in endothelial cells

Endothelial involvement has been implicated in cytomegalovirus (CMV) infection, a source of major complications in immunosuppressed individuals (e.g., those with acquired immune deficiency syndrome [AIDS] and organ transplants). Traditionally, CMV has been grown in fibroblasts; however, propagation in these cells may alter characteristics of the virus. In developing an in vitro model system of CMV/endothelial cell interaction, we have addressed this issue by propagating a clinical isolate, CMV VHL 1, in human umbilical vein endothelial (HUVE) cells by serial cocultivation of heavily infected cultures with fresh HUVE monolayers and have compared its infectious properties with those of the fibroblast-raised strain, CMV AD169. In situ hybridization using a biotinylated DNA probe, as well as immunofluorescent staining for CMV-specific antigen, has confirmed infection of HUVE cells inoculated with either strain of the virus. Infection of HUVE by VHL was accompanied by dramatic cytopathology not observed in AD169-infected cells. Plaque assay of culture supernatants revealed greater virus production in VHL-infected HUVE as compared with equivalently inoculated fibroblasts. In contrast, AD169 production in inoculated fibroblasts exceeded that in HUVE. These studies demonstrate the suitability of cultured endothelial cells as a substrate for CMV propagation and suggest that a strain of virus thus propagated may offer an accurate model of CMV/endothelial cell interaction in human disease.     

Circulating cell-derived microparticles in women with pregnancy loss

Citation
Alijotas‐Reig J, Palacio‐Garcia C, Farran‐Codina I, Zarzoso C, Cabero‐Roura L, Vilardell‐Tarres M. Circulating cell‐derived microparticles in women with pregnancy loss. Am J Reprod Immunol 2011; 66: 199–208 Problem To analyze cell‐derived microparticles (cMP) in pregnancy loss (PL), both recurrent miscarriages (RM) and unexplained fetal loss (UFL). Method of study Non‐matched case–control study was performed at Vall d’Hebron Hospital. Cell‐derived microparticles of 53 PL cases, 30 with RM, 16 with UFL, and 7 (RM + UFL), were compared to 38 healthy pregnant women. Twenty healthy non‐pregnant women act as controls. Cell‐derived microparticles were analyzed through flow cytometry. Results are given as total annexin (A5+), endothelial‐(CD144+/CD31+ CD41?), platelet‐(CD41+), leukocyte‐(CD45+) and CD41? c‐MP/μL of plasma. Antiphospholipid antibodies (aPLA) were analyzed according to established methods. Results Comparing PL versus healthy pregnant, we observed a significant endothelial cMP decrease in PL. When comparing RM subgroup with controls, we observed significant decreases in endothelial cMP. When comparing the PL positive for aPLA versus PL‐aPLA‐negative, no cMP numbering differences were seen. Conclusion Pregnancy loss seems to be related to endothelial cell activation and/or consumption. A relationship between aPLA and cMP could not be demonstrated.     

Balamuthia mandrillaris and Acanthamoeba amebic encephalitis with neurotoxoplasmosis coinfection in a patient with advanced HIV infection

We describe a patient with advanced HIV infection and Balamuthia mandrillaris and Acanthamoeba amebic encephalitis with Toxoplasma gondii coinfection. A multidisciplinary effort and state-of-the-art diagnostic techniques were required for diagnosis. Our patient is the first reported case of an HIV-infected person with dual Balamuthia mandrillaris and Acanthamoeba amebic encephalitis with neurotoxoplasmosis coinfection.     

CT影像中筛前动脉与颅底的关系及在鼻窦炎手术中的意义

目的 通过CT影像探讨筛前动脉（AEA）与颅底的解剖关系及其在鼻窦炎手术中的意义。方法 回顾性研究2017年1月～2017年8月共52例鼻窦炎患者,所有患者均行鼻窦CT扫描及重建,测量AEA与颅底的距离并分型,测量AEA至额嘴的距离,并研究其与AEA悬空的关系。记录眶上筛房（SOEC）的发生率并以卡方检验分析SOEC与AEA悬空的关系,测量筛板外侧板的深度并进行Keros分型,以 Spearman 相关系数分析Keros分型与AEA悬空的关系。结果 AEA在CT图像中辨别率为100%, Ⅰ型为AEA嵌于颅骨内,占 42.3%（44/104）,Ⅱ型为AEA紧贴颅底,占18.3%（19/104）,Ⅲ型为AEA悬空于筛窦内,占39.4%（41/104）,即AEA的悬空率为39.4%,至颅底的平均距离为（3.8±1.5）mm。AEA至额嘴平均距离（14.1±2.2）mm,其距离在AEA悬空与非悬空组中差异无统计学意义（t=0.740,P>0.05）。在Keros分型中,Ⅰ型占51.9%（54/104）,Ⅱ型占37.5%（39/104）,Ⅲ型占10.6%（11/104）。Keros分型与AEA发生悬空之间的Spearman相关系数为0.505（P<0.001),为中度正相关。SOEC发生率为17.3%（17/104）,有SOEC的患者与无SOEC的患者的AEA悬空发生率差异存在统计学意义（χ2=4.3287,P<0.05）。结论 当SOEC存在或Keros分型级别较高时,AEA的悬空率明显升高,术前进行CT影像学检查可以识别颅底解剖情况,明确AEA与颅底的位置关系,进而降低术中AEA的损伤风险。     

Lack of transmission to polymorphonuclear leukocytes and human umbilical vein endothelial cells as a marker of attenuation of human cytomegalovirus.

The development of a human cytomegalovirus (HCMV) vaccine for the prevention of perinatal disease is urgently needed. However, markers of HCMV attenuation are not defined at present. An in vitro model for the study of interactions of HCMV-infected human fibroblasts and peripheral blood polymorphonuclear leukocytes showed that (1) known laboratory-adapted HCMV strains as well as cell culture-adapted HCMV isolates were not transmitted to polymorphonuclear leukocytes; (2) each of the 80 HCMV recent isolates was consistently transmitted to polymorphonuclear leukocytes as both infectious virus and viral components (nucleic acid and proteins); and (3) all 15 polymorphonuclear leukocyte-tropic strains tested thus far were also endothelial cell-tropic. The in vitro transmissibility to polymorphonuclear leukocytes (and endothelial cells) is proposed as a surrogate marker of pathogenicity of HCMV strains. It seems reasonable to assume that a HCMV strain candidate for a vaccine be verified as deprived of the transfer property to polymorphonuclear leukocytes (and endothelial cells) before involvement in clinical trials assessing safety and immunogenicity.     

Non- pancreatic neuroendocrine tumour presenting with hypoglycemia in an elderly patient

BackgroundHypoglycemia is a common, symptom seen in individuals. Hypoglycemia in the elderly is both under-recognized and misdiagnosed due to nonspesific hypoglycemic symptoms and accompanying comorbidities in this population. In diabetic individuals, hypoglycemia is most commonly caused by administering insulin or sulphonylureas and insulin secretagogues. Other drugs, such as antibiotics or beta-blockers, have been reported to reduce blood glucose to abnormally low levels. Hypoglycemia in non-diabetic patients is considered a rare event, and the possible reasons may be reactive hypoglycemia, insulin-secreting tumours and other malignancies, hypopituitarism, hypocortisolism, alcohol abuse, inappropriate insulin self-administration, malnutrition, renal failure and sepsis.CaseAn 86- year- old male was admitted to the emergency department with hypoglycemia diagnosed with non-pancreatic neuroendocrine tumour (NET) on lung secreting insulin. No surgical intervention or chemotherapy was planned due to patients age and comorbidities so best supportive care was planned. We used prednisone for symptomatic treatment of hypoglycemia and the patient has been followed up periodically. In this period he had no hypoglycemic attack.ConclusionFor patients with hypoglycemia who are unable/decline to receive any further treatment, low dose glucocorticoid is a good choice to achieve normoglycemia. It seems to be more cost effective compared to other treatment options. Furthermore hospitalisation rates may decrease due to decreased hypogylcemic attacks.     

Qualitative and quantitative PCR measures of cytomegalovirus in patients with advanced HIV infection who require transfusions

The Viral Activation Transfusion Study (VATS) was a randomized trial that compared leukocyte-reduced transfusions with unfiltered red blood cell transfusions in HIV and cytomegalovirus (CMV) antibody-positive patients with anemia who were undergoing their first blood transfusion. The relations of the baseline qualitative and quantitative polymerase chain reaction (PCR) measures of plasma CMV viremia, HIV RNA, CD4(+) cell counts, and quality of life in these study subjects were examined. The 511 study subjects had a median CD4(+) cell count equal to 15 cells/mm3, and 110 (21.5%) had CMV viremia by qualitative assay. In multivariate models, frequency of positive qualitative CMV increased with decreasing CD4(+) cell counts (p =.04 trend), higher HIV RNA (p <.001), and a history of CMV disease (p <.001). Quantitative CMV PCR were performed on the 110 qualitative assay-positive study subjects. Median CMV viral load was 1780 copies/ml. In multivariate regression models, lower CD4(+) cell count (p =.03), and a history of CMV disease (p <.001) correlated with the level of CMV load. HIV RNA load and CMV load were not correlated. A lower Karnofsky score was associated with both the presence and quantity of CMV DNA.     

Antigen presenting cells in a non-mammalian model system, the chicken

The chicken has a different repertoire of tissues, cells and genes of the immune response compared to mammals, yet generally survives infection with viral, bacterial, protozoal and fungal pathogens, and also worms and ectoparasites, just like mammals. Poultry are also probably the most heavily vaccinated group of farmed animals. Antigen presentation to the adaptive immune response therefore obviously normally occurs efficiently in birds. Although comparatively much is known about macrophages and B cells in the chicken, there is as yet little work on the other, professional, antigen-presenting cells, the dendritic cells (DC). Birds also have at least two other sets of phagocytic cells, heterophils and thrombocytes, which may also have the ability to present antigen. Here we review the current state of knowledge about antigen presenting cells in the chicken, concentrating mainly on recent advances in our knowledge of DC.     

共培养条件下乳腺癌细胞对淋巴管内皮细胞增殖的影响

目的建立乳腺癌细胞和淋巴管内皮细胞共培养模型,探讨乳腺癌细胞在共培养条件下对淋巴管内皮细胞增殖的影响。方法培养并鉴定淋巴管内皮细胞;利用transwell小室建立乳腺癌细胞与淋巴管内皮细胞体外共培养模型;采用MTT法检测乳腺癌细胞对淋巴管内皮细胞增殖情况。结果VEGFR-3抗体对培养的细胞进行鉴定,为典型淋巴管内皮细胞;MTT法结果显示：在共培养24~72 h时,乳腺癌细胞能够促进共培养条件下淋巴管内皮细胞的增殖（P〈0.05）。结论成功建立乳腺癌细胞和淋巴管内皮细胞共培养模型,乳腺癌细胞能够促进共培养条件下淋巴管内皮细胞的增殖。     

Typical nemaline bodies presenting in a patient with polymyositis

A 66-year-old woman presented with a progressive myopathy affecting the proximal limbs and unusual pathological findings of nemaline bodies on muscle biopsy. Histological examination demonstrated that the bodies were mainly located in the subsarcolemmal region of atrophic fibers, exhibited strong immunoreactivity with antibodies to both alpha-actinin and m-actin, and had a typical lattice-like appearance at higher magnification on electron microscopy. These findings were the same as those for nemaline myopathy. The patient responded to steroid therapy, but relapse occurred after steroid was discontinued. Given the clinical criteria of polymyositis, we believe that the occurrence of nemaline bodies in our patient should be interpreted primarily as an epiphenomenon of primary myopathy.     

Circulating endothelial cells are increased in chronic myeloid leukemia blast crisis

We measured circulating endothelial precursor cells (EPCs), activated circulating endothelial cells (aCECs), and mature circulating endothelial cells (mCECs) using four-color multiparametric flow cytometry in the peripheral blood of 84 chronic myeloid leukemia (CML) patients and 65 healthy controls; and vascular endothelial growth factor (VEGF) by quantitative real-time PCR in 50 CML patients and 32 healthy controls. Because of an increase in mCECs, the median percentage of CECs in CML blast crisis (0.0146%) was significantly higher than in healthy subjects (0.0059%, P<0.01) and in the accelerated phase (0.0059%, P=0.01). There were no significant differences in the percentages of CECs in chronic- or active-phase patients and healthy subjects (P>0.05). In addition, VEGF gene expression was significantly higher in all phases of CML: 0.245 in blast crisis, 0.320 in the active phase, and 0.330 in chronic phase patients than it was in healthy subjects (0.145). In conclusion, CML in blast crisis had increased levels of CECs and VEGF gene expression, which may serve as markers of disease progression and may become targets for the management of CML.     

Nitric oxide induced by tumor cells activates tumor cell adhesion to endothelial cells and permeability of the endothelium in vitro

Human fibrosarcoma HT1080 cell surface phenotype analysis revealed the expression of "cluster of differentiation 15" (CD15) antigen and to a lesser extent, of "very late antigen-4" (VLA-4). Expression of "endothelial-leukocyte adhesion molecule-1" (ELAM-1) was negligible on resting human umbilical vascular endothelial cells (HUVECs), but its expression could be induced by HT1080 conditioned medium. HT1080 cell adhesion to HUVECs was partially dependent on CD15/ELAM-1 adhesion molecules. HT1080 cell adhesion to HUVECs induced the enhancement of nitric oxide (NO) production from HUVECs. Exogenous NO and NO from HUVECs enhanced ELAM-1 expression on HUVECs, HT1080 cell adhesion to HUVECs, permeability of the HUVEC monolayer, and HT1080 cell invasion through the HUVEC monolayer. These enhancements were not induced by NO synthase inhibitor, N^G-nitro-L-arginine methyl ester (L-NAME). These results suggest that NO expression induced by tumor cells via the CD15/ELAM-1 adhesion system may contribute to enhancement of tumor cell adhesion to endothelial cells and hyperpermeability of the endothelium, facilitating tumor cell invasion.     

Emergence of cytomegalovirus resistance to foscarnet in a patient receiving foscarnet salvage therapy for multidrug-resistant HIV infection

Foscarnet (FOS) is a pyrophosphate analogue that inhibits both cytomegalovirus (CMV) and human immunodeficiency virus (HIV) replication. We describe herein the emergence of CMV resistance to FOS in a 54-year-old man receiving FOS salvage therapy because of multidrug-resistant HIV-1 infection. Oral valganciclovir (VGCV) treatment allowed subsequently the improvement of FOS-resistant CMV infection.     

Early hematopoietic reconstitution after autologous transplantation with blood-derived stem cells in a patient with advanced lymphoma

A 30-year-old man with advanced non-Hodgkin lymphoma underwent repeated leukaphereses for harvesting blood-derived hemopoietic stem cells. Collection was started 8-10 days after the end of L-VAMP therapy (3 cycles). Nine procedures were performed and a total of 65.4 x 10(9) mononuclear cells (0.87 x 10(9)/Kg) were collected, processed, cryopreserved and stored in liquid nitrogen. The yields of CFU-GM, BFU-E and CFU-GEMM were respectively 964 x 10(4) (12.4 x 10(4)/Kg), 249 x 10(4) (3.2 x 10(4)/Kg) and 798 x 10(4) (10.4 x 10(4]. The patient received a myeloblative regimen consisting of fractionated total body irradiation (1200 cGy) and cyclophosphamide (120 mg/kg) followed by infusion of his own thawed cells. Early trilineage hematopoietic recovery was first observed on day +8; 1 x 10(9)/l WBC were reached on day +11, 0.5 x 10(9)/l PMN on day +13 and 50 x 10(9)/l platelets on day +11. Course was uneventful and the patient was discharged from hospital on day +21. Eight months after transplant the patient is in continuous unmaintained complete remission with normal blood cell counts. This reports suggests that complete and sustained engraftment can be achieved with peripheral stem cells recruited after "soft" chemotherapy.