Estrogen deficiency increases osteoclastogenesis up-regulating T cells activity: a key mechanism in osteoporosis

Compelling evidences suggest that increased production of osteoclastogenic cytokines by activated T cells plays a relevant role in the bone loss induced by estrogen deficiency in the mouse. However, little information is available on the role of T cells in post-menopausal bone loss in humans. To investigate this issue we have assessed the production of cytokines involved in osteoclastogenesis (RANKL, TNFalpha and OPG), in vitro osteoclast (OC) formation in pre and post-menopausal women, the latter with or without osteoporosis. We evaluated also OC precursors in peripheral blood and the ability of peripheral blood mononuclear cells to produce TNFalpha in both basal and stimulated condition by flow cytometry in these subjects. Our data demonstrate that estrogen deficiency enhances the production of the pro-osteoclastogenetic cytokines TNFalpha and RANKL and increases the number of circulating OC precursors. Furthermore, we show that T cells and monocytes from women with osteoporosis exhibit a higher production of TNFalpha than those from the other two groups. Our findings suggest that estrogen deficiency stimulates OC formation both by increasing the production of TNFalpha and RANKL and increasing the number of OC precursors. Women with post-menopausal osteoporosis have a higher T cell activity than healthy post-menopausal subjects; T cells thus contribute to the bone loss induced by estrogen deficiency in humans as they do in the mouse.     

Bone marrow stromal cell lines having high potential for osteoclast-supporting activity express PPARγ1 and show high potential for differentiation into adipocytes

Bone marrow stromal cells support osteoclast differentiation by expressing receptor activator of NF-kB ligand (RANKL). Although several bone marrow stromal cell lines have been established and characterized, the differentiation stage of the supporting cells for osteoclast differentiation remains unclear. We have established several bone marrow stromal cell lines from transgenic mice harboring the temperature-sensitive SV40 large T antigen. Some of these temperature-sensitive bone marrow stromal cell lines (TSB cell lines) support osteoclast differentiation and differentiate into osteoblasts, suggesting that osteoblast precursor cells support osteoclast differentiation. Here we show that the TSB cell lines that support osteoclast differentiation also expressed peroxisome proliferator-activated receptor γ1 (PPARγ1) and were able to differentiate into adipocytes. PPARγ1 is an alternatively spliced form of PPARγ that is responsible for the adipocyte differentiation and expressed in the adipocyte precursor cells. Immunofluorescence analysis of TSB cell lines and primary bone marrow stromal cells by use of anti-PPARγ and anti-RANKL antibodies showed that fluorescent signals for RANKL were observed in the cells that expressed PPARγ. Furthermore, activation of adipocyte differentiation by a PPARγ agonist led to decreased RANKL expression. These results demonstrate that PPARγ1-positive precursor cells for osteoblasts and adipocytes expressed RANKL and supported osteoclastogenesis.     

Pacientes com lúpus eritematoso sistêmico e síndrome antifosfolípide secundária possuem números reduzidos de células B CD4+ CD25+ Foxp3+ (células Treg) e células B CD3– CD19+ circulantes

IntroductionCD4+CD25+Foxp3+ regulatory T (Treg) cell depletion has been reported in systemic lupus erythematosus (SLE) and, recently, in primary antiphospholipid syndrome (APS); the issue has not been studied in SLE patients with secondary APS (SLE/APS) so far.ObjectiveTo quantify total lymphocytes, Treg cells, CD3+CD19– T cells and CD3–CD19+ B cells in SLE/APS patients and healthy controls.MethodsCell subtypes underwent immunophenotyping using specific monoclonal antibodies (anti–CD3 CY5, anti–CD4 FITC, anti–CD25, anti–Foxp3, anti–CD19 PE) and flow cytometry.ResultsTwenty–five patients with SLE/APS (mean age 43.5 years, 96% females, 96% caucasians, mean duration of disease 9.87 years, mean SLEDAI 10 ± 5.77) and 25 age and sexmatched controls entered the study. It was realized that the numbers of Treg and CD3– CD19+ B cells were significantly lower in SLE/APS patients than in controls (all p < 0.05). Treg and CD3–CD19+ B cells remained numerically low after controlling (ANCOVA) for percentage of total lymphocytes (p < 0.05). Decreasing levels of circulating Treg and CD3–CD19+ B cells correlated to higher scores of lupus activity (rs = –0.75, p < 0.0001; rs = –0.46, p = 0.021, respectively). Number of Treg cells and CD3–CD19+ B lymphocytes did not significantly differ in users or nonusers of chloroquine, azathioprine and corticosteroids (all p > 0.05).ConclusionsIn this preliminary study, patients with SLE and secondary APS showed depletion of Treg and CD3–CD19+ B cells; decreasing numbers of both subtypes correlated to a higher SLEDAI. Treg cells depletion might contribute to the autoimmune lesion seen in patients with SLE/APS. The reduced number of CD3–CD19+ B cells seen in these patients deserves more studies in order to get further elucidation.     

A comparative study of general anesthesia versus combined spinal–epidural anesthesia on the fetus in cesarean section

BackgroundObstetric anesthesia guidelines recommend regional over general anesthesia for most caesarean sections to decrease the risk for both fetus and mother.Aim of the workTo determine the effects of combined spinal epidural anesthesia and general anesthesia on the newborns and the mother undergoing elective cesarean section.SubjectsA total of 60 consecutive women with uncomplicated singleton pregnancies at term and scheduled to undergo elective cesarean section at Kasr Al-Aini obstetric hospital participated in this prospective study. The women were divided into 2 groups (each 30), a general anesthesia group (A) and combined spinal–epidural anesthesia group (B).MethodsUmbilical artery blood gas analysis and Apgar scores were assessed at 1 and 5 min after delivery in the newborn while systolic and diastolic blood pressure, heart rate, oxygen saturation and (capnography in general anesthesia) were measured preoperative and after 5, 10 and 15 min of induction of anesthesia in the mothers. In addition, the time from induction of anesthesia till delivery of the fetus and duration in operative room were measured.ResultsApgar score recorded statistically significant differences between the 2 groups at 1 min and 5 min, where with combined spinal–epidural anesthesia the Apgar score readings were higher than with general anesthesia. HCO₃ readings showed a statistically significant difference between the 2 groups after 1 and 5 min, where the newborns in general anesthesia group had a statistically significant lower HCO₃ compared to the newborns in combined spinal–epidural group. Patients in general anesthesia group were significantly more tachycardic compared to patients in combined spinal–epidural group.ConclusionCombined spinal–epidural anesthesia is safer on the newborn than general anesthesia regarding the APGAR scores and acid–base balance.     

Routine Laboratory Testing Every 4 Versus Every 6 Weeks for Patients on Maintenance Hemodialysis: A Quality Improvement Project

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Is OSNA better than imprint cytology for intraoperative diagnosis of cancer involvement of axillary sentinel node in breast cancer?

ObjectiveThe study aim was to establish Sensitivity, Specificity, Positive Predictive Value, Negative Predictive Value (NPV), and Accuracy Values of both imprint cytology (IC) and the OSNA assay for intraoperative assessment of axillary sentinel node (SN) cancer involvement in breast cancer. Specifically, we wished to find out if true positive and false negative results of IC were associated to axillary lymphadenectomy (ALND). Also, we addressed a comparative cost analysis between techniques.Methods244 patients treated for breast cancer in the Breast Unit of Hospital Germans Trias i Pujol from 2011 to 2015 were prospectively included. A transversal, consecutive design was applied to assess IC compared to the reference test (OSNA). Inclusion criteria were: T1 and T2 tumors with negative nodes, both clinically and on ultrasound.ResultsSensitivity of IC for macrometastases was 70%. The NPV of IC for macrometastases was 95,75%. Accuracy of IC was 96,12%. In the comparative cost analysis, the release time of results for OSNA doubled that of IC and was associated with an increased cost of € 370.ConclusionsIC has been stated as a good technique for intraoperative cancer involvement SN with high sensitivity and NPV compared to the OSNA assay. It allows keeping the whole node tissue and thus the possibility of improved histopathological evaluation, which can be useful for adjuvant, and offers the advantage of being less time consuming. Cost analysis shows a higher cost for OSNA, which may exceed the benefit of sorting out false negatives from IC.     

骨髓间充质干细胞对糖尿病周围神经病变大鼠骨组织OGP/RANKL表达的影响

目的检测骨髓间充质干细胞局部注射后糖尿病周围神经病变大鼠骨组织OPG/RANKL的表达水平。方法 8周龄SPF级雄性自发性2型糖尿病GK大鼠30只,高脂饲料喂养造模。造模成功后(随机血糖≥11.1 mmol/L)暴露大鼠左侧股骨,人为造成开放性骨折并给予克氏针复位内固定,将培养收集浓缩的骨髓间充质干细胞注射于骨折周围肌肉软组织,于实验后第12周、16周、20周分别取材用免疫组化、western blot测定骨质OPG、RANKL的表达。结果实验组大鼠术后第12周、16周、20周骨组织免疫组化和western blot测定OPG表达明显较对照组增高,而RANKL的表达结果正好相反。结论骨髓间充质干细胞局部移植可以促进GK糖尿病大鼠骨折后骨组织中OPG的高表达,抑制RANKL表达,从而促进糖尿病大鼠骨折后骨质的愈合。     

RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens

ObjectiveTo develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.MethodsA panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A–C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID₅₀ per μL and copies per μL, and the newly established methods were tested in clinical specimens collected in recent years.ResultsThe sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID₅₀ per μL and 10 virus copies per μL, and for the complete VP1 gene was 1 CCID₅₀ per μL and 100 virus copies per μL, using serially-diluted virus stocks of five serotypes. As a proof-of-concept, 25 serotypes were identified and complete VP1 sequences of 23 serotypes were obtained by this system among 858 clinical specimens positive for HEVs during the past eight surveillance seasons.ConclusionThis RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A–D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.     

Predictors of Cognitive Functions in Children With Sturge–Weber Syndrome: A Longitudinal Study

BackgroundSturge–Weber syndrome is often accompanied by seizures and neurocognitive deterioration, although previous studies have suggested that early functional brain reorganization may diminish the cognitive sequelae in some children with unilateral Sturge–Weber syndrome. The “rules” governing these plasticity mechanisms are poorly understood. In this study, we evaluated longitudinal changes of cognitive functioning (intelligence quotient [IQ]) and assessed the performance of clinical, electroencephalography (EEG), and magnetic resonance imaging (MRI) variables for predicting IQ in children with Sturge–Weber syndrome.MethodsThirty-three young children (mean age: 3.3 years at baseline) with unilateral Sturge–Weber syndrome underwent MRI, scalp EEG, and neuropsychology evaluation twice, with a median follow-up of 2 years. None of the children had epilepsy surgery. Longitudinal IQ changes were calculated. Seizure variables, interictal EEG abnormalities, and extent and location of MRI brain involvement were correlated with IQ assessed at follow-up.ResultsGlobal IQ showed a highly variable course with both increases and decreases over time. Lower IQ at baseline was associated with interval IQ increase. In univariate analyses, lower outcome IQ was associated with baseline EEG abnormalities (P < 0.001), young age at seizure onset (P = 0.001), high seizure frequency (P = 0.02), and early frontal-lobe involvement on MRI (P = 0.01). In multivariate analysis, EEG abnormalities at baseline remained a robust, independent predictor of outcome IQ.ConclusionsThe early trajectory of cognitive changes in children with unilateral Sturge–Weber syndrome is highly variable; children with improving IQ likely undergo effective unimpeded functional reorganization. Early onset, frequent seizures, and interictal epileptiform abnormalities on EEG likely interfere with this process resulting in poor cognitive functions. Future studies assessing interventions should target this high-risk subgroup to optimize cognitive outcome in Sturge–Weber syndrome.     

Meta-analysis of quality of life in children and adolescents with ADHD: By both parent proxy-report and child self-report using PedsQL™

Attention deficit hyperactivity disorder (ADHD) is a prevalent developmental disorder that seriously and negatively impacts a child's health-related quality of life (HRQOL). However, no meta-analysis has been conducted to examine the magnitude of impact, domains affected and factors moderating the impact. This review included nine studies that compared HRQOL of children or adolescents with ADHD with those with typical development using both child self-reports and parent proxy-reports. Seven among nine studies were meta-analytically synthesized to examine the degree of impact of ADHD on children and adolescents, parent–child discrepancy, and the moderators. The results indicate that ADHD impact a child's or adolescent's HRQOL negatively with a moderate effect in physical and a severe effect in psychosocial (i.e., emotional, social, and school) domains. Parental ratings of overall HRQOL in children or adolescents with ADHD were not significantly different from child's ratings when compared with typically developing children and adolescents. Age was negatively associated with all domains of HRQOL in children and adolescents with ADHD both by parent- and child-ratings, and the strongest effect was found in parental ratings of child's emotional HRQOL, with a moderate correlation. This meta-analysis suggests that HRQOL may be assessed in children and adolescents with ADHD both by parent proxy- and child self-reports, and that interventions may be planned accordingly. Future meta-analysis may explore how measures of HRQOL and other factors including child, parental, familiar and school characteristics influence the impact of ADHD and the parent–child agreement in children and adolescents.