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Contains fulltext : 252036.pdf (Publisher’s version ) (Closed access) BACKGROUND: Most physicians encounter pregnant women with cancer incidentally, leading to a lack of expertise or confidence to inform and treat these patients based on the most recent guidelines and expert opinions. In the Netherlands, a national multidisciplinary tumour board for cancer, infertility and pregnancy (CIP-MDT) was founded in December 2012, including 35 specialists from a variety of disciplines. This study evaluates the frequency of consultation of the CIP-MDT, the types of questions asked and the satisfaction of consulting physicians with its existence. METHODOLOGY: Of all requests to the CIP-MDT between December 2012 and June 2021, tumour type, stage, gestational age at diagnosis and recommendations were collected and analysed. For evaluating the methods of the CIP-MDT, a survey with questions regarding experiences with the CIP-MDT and its impact on treatment decisions was sent out to physicians that consulted the CIP-MDT. RESULTS: Recommendations (n = 213) concerned preferred and safest options for imaging, treatment options during pregnancy, possible effects on the child and fertility preserving options. Most frequently discussed malignancies were breast cancer (n = 66), cervical cancer (n = 34), haematological malignancies (n = 32) and melanoma (n = 21). The questionnaire was completed by 54% of the physicians (n = 50). Satisfaction with the recommendations of the CIP-MDT was high, and 94% of the physicians informed their patients about consulting the CIP-MDT and felt supported by the received recommendations. DISCUSSION: The national Dutch CIP-MDT contributes to a high level of satisfaction among physicians requesting advice. Further research should be executed to confirm that a CIP-MDT improves the outcomes for pregnant women and their children.

Age-related bone loss is a failure of balanced bone turnover and diminished skeletal mechanoadaptation. Estrogen receptors, ERα and ERβ, play critical roles in osteoprotective regulation activated by estrogen and mechanical signals. Previous studies mainly focused on ERα and showed that osteocyte-ERα (Ot-ERα) regulated trabecular, but not cortical bone, and played a minor role in load-induced cortical adaptation. However, the role of Ot-ERβ in bone mass regulation remains unrevealed. To address this issue, we characterized bone (re)modeling and gene expression in male and female mice with Ot-ERβ deletion (ERβ-dOT) and littermate control (LC) at 10 weeks (young) or 28 weeks (adult) of age, as well as their responses to in vivo tibial compressive loading. Increased cancellous bone mass appeared in the L

Type I Diabetes Mellitus (T1DM) and periodontitis have long been thought to be biologically connected. Indeed, T1DM is a risk factor for periodontal disease. With the population of diabetic individuals growing, it's more important than ever to understand the negative consequences of diabetes on the periodontium and the mechanisms. The aim of this study was to find out the early effects of T1DM on the periodontium without any experimentally induced periodontitis.We established the Streptozotocin (STZ)-induced diabetic mouse model and examined the periodontium 8 weeks later by histology, molecular and cellular assays. Micro-computed tomographic (𝜇CT) imaging and in vivo fluorochrome labeling were also employed to quantify bone volume and mineral apposition rates (MAR).The histologic appearance of epithelium tissue, connective tissue, and periodontal ligament in the diabetic condition was comparable to that of control mice. However, immune cell infiltration in the gingiva was dramatically elevated in the diabetic mice, which was accompanied by unmineralized connective tissue degeneration. Bone resorption activity was significantly increased in the diabetic mice, and quantitative 𝜇CT demonstrated the bone volume, the ratio of bone volume over tissue volume, and cementum-enamel junction to alveolar bone crest (CEJ-ABC) in the diabetic condition were equivalent to those in the control group. In vivo fluorochrome labeling revealed increased MAR and bone remodeling in the diabetic mice. Further investigation found the diabetic mice had more osteoprogenitors recruited to the periodontium, allowing more bone formation to balance the enhanced bone resorption.STZ-induced T1DM mice, at an early stage, have elevated gingival inflammation and soft tissue degeneration and increased bone resorption; but still the alveolar bone was preserved by recruiting more osteoprogenitor cells and increasing the rate of bone formation. We conclude that inflammation and periodontitis precede alveolar bone deterioration in diabetes. This article is protected by copyright. All rights reserved.

Calcium/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) regulates bone remodeling through its effects on osteoblasts and osteoclasts. However, its role in osteocytes, the most abundant bone cell type, and the master regulator of bone remodeling, remains unknown. Here we report that the conditional deletion of CaMKK2 from osteocytes using Dentine matrix protein 1 (Dmp1)-8kb-Cre mice led to enhanced bone mass only in female mice owing to a suppression of osteoclasts. Conditioned media isolated from female CaMKK2-deficient osteocytes inhibited osteoclast formation and function in in vitro assays, indicating a role for osteocyte-secreted factors. Proteomics analysis revealed significantly higher levels of extracellular calpastatin, a specific inhibitor of calcium-dependent cysteine proteases calpains, in female CaMKK2 null osteocyte conditioned media, compared to media from female control osteocytes. Further, exogenously added non-cell permeable recombinant calpastatin domain I elicited a marked, dose-dependent inhibition of female wild-type osteoclasts and depletion of calpastatin from female CaMKK2-deficient osteocyte conditioned media reversed the inhibition of matrix resorption by osteoclasts. Our findings reveal a novel role for extracellular calpastatin in regulating female osteoclast function and unravel a novel CaMKK2-mediated paracrine mechanism of osteoclast regulation by female osteocytes.

21 pages, 8 figures, 3 tables, supplementary material https://www.frontiersin.org/articles/10.3389/fmars.2021.570354/full#supplementary-material.-- Data Availability Statement: The datasets presented in this article are not readily available because the dataset is available by request to the Spanish Institute of Oceanography. Requests to access the datasets should be directed to webmaster@ieo.es, Small pelagic fish (SPF) in the western Mediterranean Sea are key elements of the marine food web and are important in terms of biomass and fisheries catches. Significant declines in biomass, landings, and changes in the age/size structure of sardine Sardina pilchardus and anchovy Engraulis encrasicolus have been observed in recent decades, particularly in the northern area of the western Mediterranean Sea. To understand the different patterns observed in SPF populations, we analyzed key life history traits [total length at age, length at maturity, gonadosomatic index (GSI), and body condition (Kn)] of sardine and anchovy collected between 2003 and 2017, from different fishing harbors distributed along a latitudinal gradient from northern to southern Spain. We used Generalized Linear Models (GLM) to estimate the length at maturity and Generalized Additive Models (GAMs) to test the relationship with environmental variables (seawater temperature, water currents, and net primary productivity). The life history traits of both species presented seasonal, interannual and latitudinal differences with a clear decline in length at age, length at first maturity, and body condition, for both species in the northern part of the study area. In the southern part, on the contrary, life history traits did not present a clear temporal trend. The environmental conditions partially explained the long-term changes in life history traits, but the selected variables differed between areas, highlighting the importance of regional oceanographic conditions to understand the dynamics of small pelagic fish. The truncated length-at-age pattern for both species with the disappearance of the larger individuals of the population could have contributed to the poor condition of small pelagic fish populations in the northern part of the western Mediterranean Sea in recent years. In the south area, recent declines in body condition for sardine and anchovy were observed and could be a possible first sign for future population declines. This study highlights the importance of understanding the trade-off between the energy invested in reproduction, maintenance and growth at seasonal and interannual level to advance our knowledge on how environmental and human pressures influence population dynamics of small pelagic fish at local and regional scales, This study was carried out within the Spanish Research project PELWEB (CTM2017-88939-R) funded by Spanish Ministry of Science, Innovation and Universities and the European Research Contract SPELMED (SC NR 02-TENDER EASME/EMFF/2016/032XXX) funded by EC EASME. Fisheries data collection has been co-funded by the EU through the European Maritime and Fisheries Fund (EMFF) within the National Program of collection, management and use of data in the fisheries sector and support for scientific advice regarding the Common Fisheries Policy (Regulation, EU 2017/1004), With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)

Introduction/Background The purpose of this study was to assess obstetric and maternal outcome of pregnant patients with diagnosis of Hodgkin lymphoma (HL) to guide physicians in clinical management. Methodology Clinical data of pregnant patients diagnosed with HL between 1969 and 2018 were collected from the registry of the International Network on Cancer, Infertility and Pregnancy (INCIP). For survival analysis of classical HL treated with an ABVD-based regimen, non-pregnant controls were selected based on stage and prognostic score at diagnosis. Results The median gestational age at diagnosis of 134 eligible patients was 20 weeks (range: 3–37). Antenatal chemotherapy was initiated in 53.7% of patients. Ten (7.5%) early pregnancies were terminated. One foetus deceased in the third trimester after three cycles of chemotherapy. In total, 120 (89.6%) pregnancies ended in a live birth. Preterm delivery was observed in 47 (40.1%) singleton pregnancies. Birth weight percentiles were lower in children prenatally exposed to oncological treatment and 17.9% were small for gestational age at birth (figure 1). Four children (3.5%) had major congenital malformations. Five-year progression-free survival (PFS) for HL during pregnancy was 82.5% and 90.9% for early (n=62) and advanced stage (n=15). Five-year overall survival (OS) was 97.3% and 100%, respectively. Although not significant, patients with early stage HL appeared to have inferior PFS compared with matched non-pregnant controls (n=62, figure 2), more clearly seen in the subgroup that initiated chemotherapy during pregnancy (n=45). OS was comparable between both groups, supporting the effectiveness of salvage therapy. For advanced stage HL survival was similar to controls, albeit small numbers. Conclusion Survival of patients diagnosed with early stage HL during pregnancy appears not statistically different from matched non-pregnant controls, however future prospective research is necessary to investigate the efficacy of chemotherapy during pregnancy. Awareness of complications as preterm delivery and low birth weight is important. Disclosure This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 647047. We are grateful to the Research Foundation-Flanders (FWO., grant no G070514N) and ESGO (European Society of Gynaecological Oncology) for their support. FA is senior clinical investigator of the F.W.O. MJH was supported by Charles University research project Progres Q28 and Q34 and by grant MH CZ - DRO (‘Kralovske Vinohrady University Hospital - FNKV, 00064173’). The funding sources did not influence study design. There are no conflicting interests to declare.

This paper considers the manufacturing process of the Oloid by means of numerical control machining. Firstly, the requirements of the end part were studied, and it was decided to manufacture the Oloid via gravity die casting, and its necessary mold to be produced by CNC machining. In order to do so, the gating system and mold had to be designed and tested using sand casting methods. When the test results were optimal the die was machined and the part cast. During this process the CAD/CAM workflow was object of observation and study, which allowed for insight and conclusions on the design process.

Background The purpose of this study was to assess obstetric and maternal outcome of pregnant patients with diagnosis of Hodgkin lymphoma (HL) registered by the International Network on Cancer, Infertility and Pregnancy (INCIP) to guide physicians in clinical management. Methods Clinical data of pregnant patients diagnosed with HL between 1969 and 2018 were collected from the INCIP registry. For survival analysis of classical HL treated with an ABVD-based regimen, non-pregnant controls were selected based on stage and prognostic score at diagnosis. Results The median gestational age at diagnosis of 134 eligible patients was 20 weeks (range: 3 – 37). Antenatal chemotherapy was initiated in 53.7% of patients. Ten (7.5%) early pregnancies were terminated. One foetus deceased in the third trimester after three cycles of chemotherapy. In total, 120 (89.6%) pregnancies ended in a live birth. Preterm delivery was observed in 47 (40.1%) singleton pregnancies. Birth weight percentiles were lower in children prenatally exposed to oncological treatment and 17.9% were small for gestational age at birth. Four children (3.5%) had major congenital malformations. Five-year progression-free survival (PFS) for HL during pregnancy was 82.5% and 90.9% for early (n = 62) and advanced stage (n = 15). Five-year overall survival (OS) was 97.3% and 100%, respectively. Although not significant, patients with early stage HL appeared to have inferior PFS compared with matched non-pregnant controls (n = 62), more clearly seen in the subgroup that initiated chemotherapy during pregnancy (n = 45). OS was comparable between both groups, supporting the effectiveness of salvage therapy. For advanced stage HL survival was similar to controls, albeit small numbers. Conclusions Although further prospective research on the efficacy of chemotherapy during pregnancy is necessary, survival of patients diagnosed with early stage HL during pregnancy appears not to be statistically different from matched non-pregnant controls, Awareness of complications as preterm delivery and low birth weight is important in this population. Clinical trial identification NTC00330447. Legal entity responsible for the study The authors. Funding European Union’s Horizon 2020 Research and Innovation Program under grant agreement No 647047 Research Foundation-Flanders (FWO., grant no G070514N) and ESGO (European Society of Gynaecological Oncology) Charles University Research Project Progres Q28 and Q34 and by grant MH CZ - DRO ("Kralovske Vinohrady University Hospital - FNKV, 00064173"). Disclosure All authors have declared no conflicts of interest.

Pseudoephedrine (PSE) is a stereoisomer of ephedrine that is commonly used as a nasal decongestant in combination with other anti-inflammatory drugs for the symptomatic treatment of some common pathologies such as common cold. Herein, we describe for the first time the effects of PSE on T-cell activation events. We found that PSE inhibits interleukin-2 (IL-2) and tumor necrosis factor (TNF) alpha-gene transcription in stimulated Jurkat cells, a human T-cell leukemia cell line. To further characterize the inhibitory mechanisms of PSE at the transcriptional level, we examined the transcriptional activities of nuclear factor kappa B (NF-κB), nuclear factor of activated T cells (NFAT), and activator protein-1 (AP-1) transcription factors and found that PSE inhibited NF-κB-dependent transcriptional activity without affecting either the phosphorylation, the degradation of the cytoplasmic NF-κB inhibitory protein, IκBα or the DNA-binding activity. However, phosphorylation of the p65/RelA subunit was clearly inhibited by PSE in stimulated cells. In addition, PSE inhibited the transcriptional activity of NFAT without interfering with the calcium-induced NFAT dephosphorylation event, which represents the major signaling pathway for its activation. NFAT cooperates with c-Jun, a compound of the AP-1 complex, to activate target genes, and we also found that PSE inhibited both JNK activation and AP-1 transcriptional activity. These findings provide new mechanistic insights into the potential immunomodulatory activities of PSE and highlight their potential in designing novel therapeutic strategies to manage inflammatory diseases.

The purpose of this study was to compare the effects of estrogen deficiency and bone mass loss on normal knee cartilage in mature rabbits. Bilateral ovariectomy (OVX) was performed in 13 rabbits, 6 of which also received systemic glucocorticoid for 4 weeks. Seven additional healthy rabbits were used as controls. Bone mineral density (BMD) was measured by dual X-ray absorptiometry in lumbar spine, knee, and subchondral bone of the knee at baseline and 22 weeks after OVX. After sacrifice, the knees were dissected, macroscopy was assessed, and histological cartilage abnormalities were evaluated according to the Mankin score. Correlations of Mankin with BMD at different regions were also performed. When compared to baseline, differences in BMD were only found in spine and knee of the animals receiving glucocorticoids. All the animals subjected to OVX had a significantly higher Mankin score than controls. Mankin was upper in OVX animals receiving glucocorticoids, but differences were not significant. The Mankin score was inversely related with BMD in lumbar spine (r = -0.67; p < 0.01). Although low bone mineral density contributes to the minor osteoarthritic alterations observed in our model, estrogen deficiency itself seems to act directly to induce the main pathogenic effects in healthy cartilage of the rabbit.

[EN]: Background and objective: Hypertensive disorders of pregnancy could be favoured by polymorphisms in genes affecting vascular physiology. The aim of our work was to study several variants in the genes regulating oxidative stress, plasma lipids metabolism and endothelial function (observational study). Material and methods: We studied the −930A/G polymorphism of the CYBA gene promoter, the apolipoprotein E (APOE) genotype and the methylene-tetrahydrofolate reductase (MTHFR) gene C677T polymorphism in 134 healthy pregnant women, 266 pregnant with non-proteinuric hypertension (NPH) and 184 patients with preeclampsia (PE). Results: The GG genotype of the CYBA gene promoter was present in 32.1% of the control population, 38.7% of patients with NPH (P=0.19) and 21.2% of the women with PE (P=0.03). A higher frequency of 3/ 4 and 4/ 4 genotypes of APOE was observed in patients with PE or NPH compared with controls (P, [ES]: Fundamento y objetivo: Nos propusimos valorar en un estudio observacional si algunos polimorfismos en genes que regulan el estrés oxidativo, los niveles de homocisteina y el metabolismo de los lípidos, podrían predisponer a diferentes trastornos hipertensivos del embarazo. Material y métodos: Estudiamos el polimorfismo −930A/G del gen promotor del CYBA, el genotipò de la apolipoproteína E (ApoE) y el polimorfismo C677T del gen de la metilen tetrahidrofolato-reductasa (MTHFR) en 134 embarazadas sanas, 266 embarazadas con hipertensión no proteinurica (HNP) y 184 pacientes con preeclampsia (PE). Resultados: El genotipo GG del promotor del gen del CYBA estuvo presente en el 32,1% de la población de control, en el 38,7% de las pacientes con HNP (p=0,19) y el 21,2% de las mujeres con PE (p=0,03). Los pacientes con PE o HNP, en comparación con los controles, mostraron una mayor frecuencia de genotipos 3/ 4 y 4/ 4 (p

Increased fracture risk is commonly reported in cancer patients receiving radiotherapy, particularly at sites within the field of treatment. The direct and systemic effects of ionizing radiation on bone at a therapeutic dose are not well-characterized in clinically relevant animal models. Using 20-week-old male C57Bl/6 mice, effects of irradiation (right hindlimb; 2 Gy) on bone volume and microarchitecture were evaluated prospectively by microcomputed tomography and histomorphometry and compared to contralateral-shielded bone (left hindlimb) and non-irradiated control bone. One week postirradiation, trabecular bone volume declined in irradiated tibias (-22%; p 0.0001) and femurs (-14%; p = 0.0586) and microarchitectural parameters were compromised. Trabecular bone volume declined in contralateral tibias (-17%; p = 0.003), and no loss was detected at the femur. Osteoclast number, apoptotic osteocyte number, and marrow adiposity were increased in irradiated bone relative to contralateral and non-irradiated bone, whereas osteoblast number was unchanged. Despite no change in osteoblast number 1 week postirradiation, dynamic bone formation indices revealed a reduction in mineralized bone surface and a concomitant increase in unmineralized osteoid surface area in irradiated bone relative to contralateral and non-irradiated control bone. Further, dose-dependent and time-dependent calvarial culture and in vitro assays confirmed that calvarial osteoblasts and osteoblast-like MC3T3 cells were relatively radioresistant, whereas calvarial osteocyte and osteocyte-like MLO-Y4 cell apoptosis was induced as early as 48 hours postirradiation (4 Gy). In osteoclastogenesis assays, radiation exposure (8 Gy) stimulated murine macrophage RAW264.7 cell differentiation, and coculture of irradiated RAW264.7 cells with MLO-Y4 or murine bone marrow cells enhanced this effect. These studies highlight the multifaceted nature of radiation-induced bone loss by demonstrating direct and systemic effects on bone and its many cell types using clinically relevant doses; they have important implications for bone health in patients treated with radiation therapy.

The case is reported of acute retinal necrosis with bilateral involvement due to Varicella Zoster virus in a 77 year-old man. Polymerase chain reaction (PCR) of aqueous humor was positive for Varicella Zoster virus (VZV). He developed a Kyrieleis' vasculitis a month after the starting treatment, when the PCR analysis was negative.PCR is a quick and safe technique, with a high sensitivity and specificity of 97%, useful to diagnose and monitor the viral activity. The intervention must be urgent, due to the dramatically rapid evolution. Oral famciclovir oral is good alternative owing to its bioavailability.

Aim : To determine the efficacy of flow cytometry (FC) in the assessment of bone marrow (BM) in B-cell non-Hodgkin lymphoma (B-NHL). FC is a common practice, but is far from being validated. Methods and results : Morphological analysis and FC immunophenotyping were perfomed on 421 samples. T-cell lymphomas, Hodgkin's disease, chronic lymphocytic leukaemia and hairy cell leukaemia were not included in the study. Clonality was assessed by the standard κ/λ/CD19 test. Aberrant immunophenotypes present in the B-cell subpopulation were also investigated. A double-step procedure was employed in all cases to increase the sensitivity of the FC procedure. Of 380 evaluable samples, 188 corresponded to follicular lymphoma (FL), 58 to diffuse large B-cell lymphoma (DLBCL), 57 to mantle cell lymphoma (MCL), seven to Burkitt's lymphoma and the remaining 70 samples to other low-grade lymphomas. Morphological marrow infiltration was found in 148 cases, and flow immunophenotyping identified 138 cases with BM involvement. A concordance between the two methods was detected in 298 cases (79%). There was a discordance in 82 cases (21%): morphology positive/FC negative in 46 cases and morphology negative/FC positive in 36 (61% of all cases with discordance were from FL). There was no difference in outcome when patients with discordances were compared with patients without discordances. Conclusions : Most samples showed concordance between morphological and FC results. FC identified BM involvement in the absence of morphological infiltration. Morphology/FC discordance seems to have no influence on the outcome of FL patients.

We report a case of simultaneous diagnosis of chronic lymphocytic leukemia (CLL) and acute myeloid leukemia (AML), in which the use of flow cytometry analysis allowed the demonstration of two different cell populations and the study of both immunophenotyping patterns with a large panel of monoclonal antibodies (MoAbs). CLL cells showed a typical immunophenotype with coexpression of B cell markers with CD5, CD23, CD43, and weak surface immunoglobulin light chain restriction expression, whereas the AML population had a very uncommon phenotype with expression of myeloid markers and CD56 and lack of expression of other natural killer (NK) antigens, CD34 and HLA-DR. After chemotherapeutic treatment of AML with two induction courses, the patient achieved complete remission of the AML with persistence of a CD19/CD5 positive population. After consolidation chemotherapy, this latter population was no longer detectable despite the presence of lymphoid nodules in a bone marrow biopsy. Six months after diagnosis, the patient relapsed with AML and died shortly afterwards.

Altered bone turnover is a key pathologic feature of chronic kidney disease-mineral and bone disorder (CKD-MBD). Expression of TGF-β1, a known regulator of bone turnover, is increased in bone biopsies from individuals with CKD. Similarly, TGF-β1 mRNA and downstream signaling is increased in bones from jck mice, a model of high-turnover renal osteodystrophy. A neutralizing anti-TGF-β antibody (1D11) was used to explore TGF-β's role in renal osteodystrophy. 1D11 administration to jck significantly attenuated elevated serum osteocalcin and type I collagen C-telopeptides. Histomorphometric analysis indicated that 1D11 administration increased bone volume and suppressed the elevated bone turnover in a dose-dependent manner. These effects were associated with reductions in osteoblast and osteoclast surface areas. Micro-computed tomography (µCT) confirmed the observed increase in trabecular bone volume and demonstrated improvements in trabecular architecture and increased cortical thickness. 1D11 administration was associated with significant reductions in expression of osteoblast marker genes (Runx2, alkaline phosphatase, osteocalcin) and the osteoclast marker gene, Trap5. Importantly, in this model, 1D11 did not improve kidney function or reduce serum parathyroid hormone (PTH) levels, indicating that 1D11 effects on bone are independent of changes in renal or parathyroid function. 1D11 also significantly attenuated high-turnover bone disease in the adenine-induced uremic rat model. Antibody administration was associated with a reduction in pSMAD2/SMAD2 in bone but not bone marrow as assessed by quantitative immunoblot analysis. Immunostaining revealed pSMAD staining in osteoblasts and osteocytes but not osteoclasts, suggesting 1D11 effects on osteoclasts may be indirect. Immunoblot and whole genome mRNA expression analysis confirmed our previous observation that repression of Wnt/β-catenin expression in bone is correlated with increased osteoclast activity in jck mice and bone biopsies from CKD patients. Furthermore, our data suggest that elevated TGF-β may contribute to the pathogenesis of high-turnover disease partially through inhibition of β-catenin signaling.

Introduction Dental implant osseointegration can be impaired in medical conditions with low bone mass, such as glucocorticoid-induced osteoporosis. Intermittent human parathyroid hormone (PTH) [1-34] administration has shown relevant anabolic bone activity in various animal models of osteoporosis. Therefore, we studied the effects of intermittent PTH [1-34] on bone response around titanium implants in experimental osteoporosis induced by ovariectomy and glucocorticoid administration. Methods Titanium dental implants were placed in the proximal tibia metaphysis in 38 animals. Twenty-eight rabbits had undergone bilateral ovariectomy and further methylprednisolone administration for 4 weeks to induce osteoporosis. Ten healthy rabbits were used as controls. At week 8, osteoporotic rabbits started saline vehicle or intermittent PTH administration for 12 weeks. Bone mineral density (BMD) was assessed in peri-implant area, lumbar spine, and global and subchondral knee bone at baseline, and weeks 6 and 20. Animal sacrifice was carried out at week 21. Afterward, tibiae were removed for μCT morphometry and undecalcified sections were evaluated by light and scanning electron microscopy. Results PTH increased bone-to-implant contact compared with control rabbits or vehicle administration in osteoporotic rabbits (P

SummaryPurposeSynoviopathy contributes to cartilage degradation in osteoarthritis (OA). Intermittent parathyroid hormone (PTH) [1-34] administration inhibits terminal differentiation of human chondrocytes and prevents cartilage damage. We aimed to determine whether PTH [1-34] could modify synovial changes in experimental OA preceded by osteoporosis (OP).MethodsTwenty osteoporosis (OP) rabbits underwent knee surgery to induce OA. They were administered either saline vehicle or PTH for 10 weeks. Ten healthy rabbits were used as controls. Following sacrifice, synovial changes were assessed by Krenn synovitis score, immunohistochemistry for macrophages (RAM-11), B and T lymphocytes, type I collagen, parathyroid hormone 1 receptor (PTH1R), and anti-proliferating cell nuclear antigen (PCNA). Synovial mRNA levels of Col1A1, IL-1β, cyclooxygenase 2 (COX-2), matrix-degrading metalloproteinases (MMP-9, MMP-13), and monocyte chemotactic protein-1 (MCP-1), as well as protein expression of PTH1R were also determined. Cartilage damage was analyzed by Mankin score.ResultsOPOA + vehicle rabbits showed an increase in synovitis score vs controls (P = 0.003), mainly due to synovial hyperplasia and fibrosis, while PTH reduced these changes (P = 0.017). Mankin and Krenn scores were well correlated in all groups (r = 0.629, P = 0.012). Immunostaining for RAM-11 and B lymphocytes was increased (P ≤ 0.05), whereas PTH1R protein levels tended to be higher in OPOA + vehicle animals vs controls. PTH did not modify RAM-11 staining or PTH1R levels; however, it restored PTH1R localization to the vicinity of synovial vessels. PTH also decreased type I collagen, MCP-1, and MMP-13 expression (P

The aim of this study was to assess the importance of immunohistochemical markers in the diagnosis of pigmented conjunctival lesions. Due to the difficulty of making an exact clinical diagnosis, the suspicion of malignancy requires the removal of the lesion and performing a histopathology study in which immunohistochemical markers may help to determine the nature of the lesion.A case is presented of a 25 year-old woman with a pigmented lesion in the caruncle. It appeared recently and was growing fast with increasing pigmentation. Due to a suspicion of malignancy, the total lesion was removed. The microscopic study revealed cellular alterations which suggested malignancy. However, after carrying out immunohistochemical markers the diagnosis was conjunctival compound nevus.

Intermittent parathyroid hormone (PTH) administration has been shown to be a promising therapy for systemic bone loss. Accordingly, we hypothesized that PTH could have positive results in treating oral complications of osteoporosis. Hence, we evaluated both mandibular bone loss and its response to PTH in a rabbit model of osteoporosis induced by ovariectomy and glucocorticoid administration. There was a significant and marked decrease in bone mineral density (BMD), bone mineral content (BMC), and calcium content in ash from the osteoporotic peri-alveolar region, which influenced global jaw loss. Remarkably, PTH (1–34) administration to osteoporotic rabbits almost completely reversed BMD, BMC, and calcium content fall in the peri-alveolar region, subsequently reducing global mandibular bone loss. Thus, although the peri-alveolar region is particularly susceptible to osteoporosis, it also responds well to intermittent PTH. Therefore, these results suggest that PTH might represent a valid therapy for improving the osseointegration of dental implants in persons with osteoporosis.

Objectives. Synoviocytes play a crucial role in the inflammatory response leading to structural damage in OA. Our aim was to assess the effects of diacerein and NSAIDs on cellular responses of synoviocytes associated with inflammation and structural integrity of cartilage in OA. Methods. The effects of diacerein, celecoxib, diclofenac, meloxicam and indomethacin on prostaglandin (PG) E2 production, cyclooxygenase-2 (COX-2) protein expression, nitrite levels, presence of MMP-1 and -13, and activation of nuclear factor-� B (NF-� B) were studied on stimulated OA synoviocytes and chondrocytes. Results. Diacerein and NSAIDs inhibited IL-1� -stimulated NF-� B activation in synoviocytes and chondrocytes except indomethacin in synoviocytes. Diacerein further increased COX-2 protein expression and PGE2 synthesis in synoviocytes stimulated with IL-1� , while no effect was observed on stimulated chondrocytes. NSAIDs diminished until almost basal levels PGE2 release in both cells and, surprisingly, these drugs also diminished COX-2 protein expression both in synoviocytes and chondrocytes. With regard to structural mediators, diacerein decreased MMP-13 levels in synoviocytes but did not modify MMP-1 presence. NSAIDs induced a significant increase in MMP-1 levels in both cell types and in MMP-13 levels in chondrocytes. Conclusions. Diacerein does not seem to reduce but rather increase inflammatory mediators in synoviocytes, while it does not overall affect chondrocyte inflammatory profile.

Dear Editor, We would like to discuss the treatment of paroxysmal nocturnal hemoglobinuria (PNH) by presenting the fatal outcome of one of our patients with this disease. A 30-year-old Caucasian man with a history of auto-immune hepatitis developed pancytopenia. He had no complaints and did not take any medication. On physical examination, a pale man was seen without any other remarkable findings. His hemoglobin was 8.3 g/dL (13.8–16.9 g/dL), leucocytes 0.30 (neutrophils 0.2) × 109/L (3.5–10 × 109/L), platelet count 16 × 109/L (150–350 × 109/L) and reticulocytes 8.4 × 109/L (30–95 × 109/L) without signs of hemolysis. Bone-marrow biopsy showed reduced cellularity (10 %) without any other (including cytogenetic) abnormalities. In peripheral blood, glycosylphosphatidylinositol (PI)-deficient clones were detected in 34, 27, and 1 % of the granulocyte, monocyte, and erythroid lineages, respectively. Severe aplastic anemia (SAA) was diagnosed secondary to PNH. Lacking an HLA-identical family donor, standard treatment with immunosuppression was started: rabbit anti-thymocyte globuline, methylprednisolone, and cyclosporine. This resulted in a partial response 3 months later: persistent transfusion-dependent anemia without signs of intravascular hemolysis, thrombocytopenia, and a normal leucocyte count. An expansion of the PI-deficient clones up to 86, 86, and 7 % of the granulocyte, monocyte, and erythroid lineages, respectively (Fig. 1) was noted. Fig. 1 PNH clones at diagnosis (a) and their expansion (b) after treatment for SAA. Comparison of the PI-deficient clones at diagnosis of severe aplastic anemia (a) and after triple immunosuppression (b). Monocytes and granulocytes were gated based on scatter ... Three months after initial presentation, he was re-admitted to hospital because of abdominal pain and bloody stools. Thrombosis of the superior mesenteric vein was diagnosed and anticoagulant therapy was instituted with nadroparin 15,200 IU/day subcutaneously. Eculizumab treatment was considered, but because of the complete absence of intravascular hemolysis, the effect of the anticoagulant therapy was awaited. Subsequently, he developed headache and a left hemiparesis. Repeated CT scans of the brain showed expanding ischemic areas parieto-occipital with secondary bleeding, and the patient finally died. Thrombosis is the most feared and life-threatening complication in PNH patients, because after onset, it is frequently progressive and refractory to anticoagulant therapy [1, 2]. The mechanism of thrombotic complications in PNH is not completely understood, but it has mainly been attributed to intravascular hemolysis [1]. Experimental studies show that the free plasma hemoglobin liberated during hemolysis scavenges nitric oxide (NO). As a result, NO depletion produces dystonia and spasm of the smooth muscle, suppresses the anti-inflammatory effect on the endothelium and promotes platelet activation and thrombosis [3]. Eculizumab is a monoclonal antibody that binds complement factor C5, thereby inhibiting complement activation on erythrocytes and reducing hemolysis. As a result, eculizumab decreases thrombotic risks in PNH patients [2, 4, 5]. Interestingly, our patient suffered from progressive abdominal and cerebral thrombotic complications without any signs of intravascular hemolysis. It is also possible that the pathogenesis of thrombosis in PNH might be caused by other mechanisms associated to complement activation and not directly related to hemolysis [1, 6]. We speculate that the mechanism of thrombosis could be associated to platelet activation caused by the absence of CD59 and CD55 on the platelet surface [7, 8]. Internationally accepted indications for eculizumab treatment include transfusion-dependent hemolysis (four or more transfusions in 12 months) and PNH-related complications (i.e., thrombosis or renal failure) regardless of transfusion history [3]. Although no clinical data are available, eculizumab could be indicated in patients with thrombotic events in the absence of hemolysis [5, 9, 10].

Follicular lymphoma (FL) is a specific entity defined by characteristic histology, phenotype and molecular rearrangements. Classically, reactivity for CD19, CD10, and strong positivity for the surface light chain immunoglobulin (SIg) are considered to be phenotypic signs typically expressed in FL. In practice, this pattern is difficult to identify since most neoplastic cells analysed by flow cytometry (FC) show weak intensity for CD19-Pe/Cy5 and for SIg and negativity for CD10-FITC. We used triple antigen combinations including two monoclonal antibodies (MoAbs) against CD10 (CD10-FITC and CD10-Pe/Cy5) and a long-distance polymerase chain reaction (PCR) approach to establish the phenotypic pattern of neoplastic cells carrying t(14;18)(q32;q21). Neoplastic cells showed the following immunophenotype: stronger reactivity against CD20 than against CD19, positivity for CD22 and SIg and negativity for CD5, CD11c and CD10-FITC. Characteristically, CD10-Pe/Cy5 was expressed in all the samples with positive bcl-2/JH rearrangements. In FL, there was a high correlation between histologic diagnosis and reactivity against CD10-Pe/Cy5 (96% cases). In diffuse large cell lymphomas (DLCL), CD10-Pe/Cy5 identified positive cases with t(14;18)(q32;q21) chromosomal translocation, whereas Burkitt lymphomas showed all cases reactivity against CD10-Pe/Cy5. In conclusion, CD10-Pe/Cy5 is a useful antibody for identifying neoplastic cells carrying t(14;18)(q32;q21) in FL and DLCL. In combination with other MoAbs, anti-CD10 (HI10a, Cy-Chrome) can be used to identify a characteristic phenotypic profile of FL against other lymphoproliferative disorders.

Antibodies against CD66 identify antigens from the carcinoembryonic antigen (CEA) family of proteins, which belong to the immunoglobulin gene superfamily. Despite being usually restricted to cells of myeloid or monocytic origin, CD66 expression has also been reported in blasts from children with B-cell lineage acute lymphocytic leukaemia (ALL). An analysis of the CD66 expression was undertaken in a series of acute leukaemia patients. Antigenic expression was analysed using triple combinations of monoclonal antibodies (mAbs) in forty-five patients. The CD66 Kat4 fluorescein isothiocyanate clone was purchased from Dako (Glostrup, Denmark). CD66 was expressed in 2 of 29 patients with AML (acute myeloblastic leukemia) (6.8%) and in 8 of 12 patients with B-cell lineage ALL (66.7%; P

Acute graft-versus-host disease (AGVHD) severity is usually graded (grades 0-IV) by the pattern of organ involvement using the classic Glucksberg-Seattle criteria (GSC). Recently, the International Bone Marrow Transplant Registry (IBMTR) developed a new Severity Index by regrouping the patterns of organ involvement into five Indexes (0-D) that appeared more predictive of transplant-related mortality (TRM) and transplant failure (TF, relapse or TRM). We studied the predictive value of both grading systems of TRM, TF and GVHD-related mortality (GTRM) in a series of 114 consecutive patientsor = 12 years old allografted from a histocompatible sibling at our institution, 100 of whom were evaluable for AGVHD. The IBMTR Severity Index showed better incremental prediction of TRM (relative risks (RR) of 1, 1.5, 1.4, 2 and 2.5 for Indexes 0, A, B, C and D), TF (RRs of 1, 1.6, 1.6, 2 and 2.3, respectively) and GTRM (RRs of 1, 2.2 and 4.8 for Indexes B, C and D) than the GSC. With the GSC different outcomes for TRM and TF were found only from grade 0 to I-II and 0 to IV or I-III to IV, but not from I-II to III. The GSC also appeared less predictive of GTRM (RRs of 1, 0.4 and 2.9 for grades II, III and IV). In our relatively small patient sample, the new IBMTR Severity Index appeared more predictive of transplant outcome than the GSC, especially between no AGVHD, early Indexes (A-B) and advanced Indexes (C-D).

Electrical vehicles (Zero Emission Vehicles) represent a very elegant solution in order to decrease pollution in industrialised cities. To meet the pollution requirements, it is envisaged to replace gasoline engines by light fuell cells. They necessitate good electroconductive carbon monolithic electrodes. Monolithic organic resorcinol-formaldehyde aerogels (carbogels) can easily be made using the sol to gel method accompanied by the supercritical drying process using liquid CO 2 (critical temperature = 31 °C). Thereafter, these copolymer monolithic carbogels are pyrolysed in an atmosphere of N 2 at 1050 °C for 3 hours, in order to obtain monolithic carbon carbogels. The carbon carbogels are impregnated by H 2 PtCl 6 to obtain a weight % of metal equal to 0.44. In this work, the unique solvent used was acetone in each step, - the sol to gel reactions were catalysed by perchloric acid, from the copolymer synthesis up to the final impregnation of the carbon monoliths. The advantages of this new preparation method are compared to the aqueous former one.

We report a case of simultaneous diagnosis of chronic lymphocytic leukemia (CLL) and acute myeloid leukemia (AML), in which the use of flow cytometry analysis allowed the demonstration of two different cell populations and the study of both immunophenotyping patterns with a large panel of monoclonal antibodies (MoAbs). CLL cells showed a typical immunophenotype with coexpression of B cell markers with CD5, CD23, CD43, and weak surface immunoglobulin light chain restriction expression, whereas the AML population had a very uncommon phenotype with expression of myeloid markers and CD56 and lack of expression of other natural killer (NK) antigens, CD34 and HLA-DR. After chemotherapeutic treatment of AML with two induction courses, the patient achieved complete remission of the AML with persistence of a CD19/CD5 positive population. After consolidation chemotherapy, this latter population was no longer detectable despite the presence of lymphoid nodules in a bone marrow biopsy. Six months after diagnosis, the patient relapsed with AML and died shortly afterwards.

We studied erythropoiesis in 31 patients with vitamin B12 deficiency by measuring serum erythropoietin (s-Epo), serum transferrin receptor (s-TfR, taken as an index of total erythroid activity), reticulocyte count, and the reticulocyte maturation index (RMI). s-Epo and s-TfR were measured with commercial immunoassays, whereas reticulocyte count and RMI were determined by flow cytometry. s-Epo (123 +/- 196 U/L) and s-TfR (4.1 +/- 2 mg/L) levels were increased in patients with vitamin B12 deficiency. The absolute reticulocyte counts were decreased (29 +/- 18 x 10(9)/L) with a relative increase in the most immature fractions (RMI: 29.6 +/- 18%). A significant negative relationship was found between s-Epo and Hb level (r = -0.65, p0.0001). On the average, however, s-Epo was inappropriately low for the degree of anemia, since the observed/predicted (O/P) s-Epo ratio was 0.80 +/- 0.28 in vitamin B12 deficiency vs 1.00 +/- 0.16 in a group of patients with iron deficiency anemia. It is concluded that at least a portion of patients with vitamin B12 deficiency have serum erythropoietin levels that are inappropriately low for the degree of anemia.

Introduction It's known that adherence to treatment is a key factor in the treatment of addictions. The presence of comorbid substance dependence disorder and other psychiatric disorder is very high. Comorbid psychiatric disorders interfere with adherence to drug treatment and detoxification. Aims To analyze the percentage of patients who had a voluntary discharge to Hospital Detoxification Unit and to describe sociodemographic, clinical and diagnostic test characteristics. Material and methods We descriptively studied drug dependents patients admitted to our Detoxification Unit from June 2008 to August 2012. Data was gathered at admission on demographic (age), clinical (main abused drug, psychiatric comorbidities, polydrug users, binge consumption previous intake) and alcoholtest and/or urinalysis. Results from patients with and without voluntary discharge were compared. Results The study sample included 469 patients (77.7% men, average age 38.3 ± 9). 10.7% of the patients had voluntary discharge. We found significant differences between voluntary discharge and the non-voluntary discharge group on younger people (37,6 vs 40,42,p Conclusions Few patients had a voluntary discharge. Younger people, opiate dependence, having psychotic and borderline personality co morbidity, binge consumption previous intake and positive urinalysis might be considered as risk factors for voluntary discharge.

A total of 1068 eels were examined from a population located in the Guadalquivir river estuary (37°N, 6°25′W). Maximum ages recorded were 4 + (males) and 7+ (females), and maximum lengths were 39-1 cm (males) and 54.1 cm (females). No growth was recorded between November and April, most occurring in May and, to a lesser extent, in June-October. Females grew to be larger than males. A classification analysis, based on 17 different European eel populations revealed that populations in brackish waters grew faster than those in fresh waters, but latitude also had an influence. Length-weight relationships obtained for three eel categories (males, females and undifferentiated) were used to estimate relative condition: condition cycles were similar between sexes, with increases in autumn and decreases in winter. There were monthly fluctuations in the sex ratio, and females dominated significantly in the combined catch (234 males/276 females).

SUMMARY. 1. Age, growth and reproduction in an Atlantic population of the small fish, Aphanius iberus (Cuv. & Val., 1846), from the estuary of the Guadalquivir river were studied. 2. The growth period was from April to September and the annulus on the scales was formed in April. 3. The life-span was short with a winter age-structure of 95.9% 0+ fish, 3.9% 1+ and 0.2% 2+. 4. The fecundity of this stock, before the beginning of reproduction, is represented by the formula: Fec=0.907 TL (cm)4.099. During the reproductive period 1+ females contained more, smaller eggs g−1 of gonad than 0+ fish. 5. Males matured before females, but females were larger than males. The overall sex ratio did not differ from 1:1 but males dominated significantly during most of the spawning period. 6. Aphanius iberus spawned intermittently during the reproductive period. From April to July, 1+ group specimens (7–12 months old) reproduced. Their offspring (0+ group; 3–4 months old) reproduced from July to September. Very few of the 1+ group specimens survived to spawn the following year. 7. Mean gonadosomatic indices were much higher in the 1+ group than in the 0+ group and in females compared to males. 8. The loss of condition and the disappearance of the 1 + group during spawning, may indicate the cost of a prolonged high level of reproductive effort. 9. In this productive environment, the life history of Aphanius iberus is similar to that of Gambusia affinis (Baird & Girard, 1853) and Fundulus heteroclitus (Linnaeus) found in the same area.

This study was based on examination of 1476 barbels from a first-order stream located in the Guadalquivir River basin (38°N, 4°43′W). This stock comprised nine age groups of male and 11 of females. No growth was recorded between October and March, most occurring in April–June and, to a lesser extent, in July–September. A classification analysis revealed that this stock had the lowest growth rate of 37 different European barbel populations. Length–weight relationships were obtained for 12 barbel categories and used to estimate both condition and instantaneous growth rate. Relative condition (before and after subtracting gonad weight) was similar in both sexes and was affected by gonad growth and environmental summer conditions. Spawning occurred during the second half of May (15 May is suggested as the birthday). Gonads began to develop in September (females) and in February (males). Males matured at between 7 and 9 cm fork length (f.l.) (2–4 years old) and females between 11 and 16 cm (6–7 years). The fecundity of this stock was represented by the formula F=6.07 10 4 f.l. (mm)3.0667. Larger and older fish showed a higher fecundity and bigger eggs. The overall sex ratio did not differ from 1:1.

The age, growth and reproduction of an Atlantic population of Atherina boyeri is discussed on the basis of an examination of 2510 specimens from the Guadalquivir river (37°N, 6°25′W). The life-span was short with a winter age structure of 80·4% in 0+ group, 13·2% in 1 + group and 6·4% in 2+ group. Growth was continuous throughout the year except at the peak of the spawning period (March–April). The formation of annuli extended from April to July. The condition cycles (before and after removing gonad weight) were similar for both sexes, with a minimum in May and a maximum in July. The maximum average Gonadosomatic Index (GSI) in females (8%) was twice as high as in males (4%). One-year-old males and females achieved maturity during the spawning season of the following year at an average of 40 mm FL (fork length). A. boyeri successively spawned from March to June with a maximum in April. Longer specimens spawned earlier (January). The fecundity (Fec) of this stock, before the beginning of reproduction, was represented by the formula Fec=0·865 FL (cm) 2·486 . The overall gender ratio did not differ significantly from 1:1. In this unstable and productive environment A. boyeri displayed the same life-history tactics as Aphanius iberus, Gambusia affinis and Fundulus heteroclitus , three small fish species found in the same area and characterized by fast growth rates, early maturity, frequent spawning over a long breeding season and reduced longevity.

Weight-age data from 50 Retinta beef cows from 8 to 97 mo of age located in southwestern Spain were fitted to von Bertalanffy, Brody, and Richards functions to determine the relationship between growth curve parameters and cow efficiency. Only cows having at least 31 weights were included in the analysis. Von Bertalanffy, Brody, and Richards functions were fitted to weights of each cow. Relevant parameters of the three functions are A and K, associated with the asymptotic mature weight and rate of maturing, respectively. Criteria for comparisons among the three functions were computing difficulty, goodness of fit, and lack of bias of A. Productivity indicators were number of calves weaned during the first five calving seasons (NC), average birth weight (BWT), average weaning weight (WW), and average weaning weight per cow per year (WWY). The von Bertalanffy function was selected as the most appropriate. Least squares means for A and K were 650 +/- 8.17 kg and .038 +/- .001 mo-1, respectively. The values of NC, BWT, WW, and WWY were 4.0 +/- .11 calves, 38.2 +/- .4 kg, 218 +/- 5 kg, and 172 +/- 5 kg, respectively. Regression analysis for A indicated a decrease in NC when mature weight increased (P less than .05). There was a nonsignificant trend for heavier cows (higher A) to have calves with heavier BWT or WW. The value of WWY increased (P less than .05) with increased maturing rate (K) of cows. No significant associations were found between K and BWT or WW.(ABSTRACT TRUNCATED AT 250 WORDS)

In this paper, we report our experience of lymphocyte phenotyping of a series of 108 consecutive samples using a simple flow cytometry test (Lymphogram). The kit consists of a combination of 5 different markers conjugated with three fluorochromes (CD8-FITC, CD19-FITC, CD56-PE, CD3-PE, CD4-PECy5) in the same tube. This allows identification of different T-cells, NK subpopulations and B lymphocytes.The samples were divided into three groups: samples with absolute lymphocytosis (5 x 10(9)/L) (n = 50), samples with relative lymphocytosis (50%) (n = 24) and other categories for which a lymphocyte immunophenotype was required (T-cell lymphoma and estimation of blood involvement in chronic lymphoproliferative disorders (CLPD) (n = 34). When CD19+ cells exceeded the normal range or there was a suspicion of CLPD without B-cell lymphopenia, clonality was investigated by means of light chain restriction analysis.In the first group, 29 samples were abnormal (10 CLPD, 3 polyclonal B-cell lymphocytosis, 13 inversions of the CD4/CD8 ratio and 3 cases with CD4 lymphocytosis) and 21 samples were regarded as normal. In the second group 7 samples showed abnormalities (2 CLPD, 3 inverted CD4/CD8 ratios and 2 with a relative increase in CD4 cells). In one sample from the third group B-cell clonality without lymphocytosis was detected whereas in 18 samples a polyclonal pattern was observed. The presence of B-cell lymphopenia precluded further clonality study in 13 samples.Lymphogram associated with clonality analysis is a rapid, easy and cheap method of assessing lymphocyte phenotypes in the majority of clinically relevant situations.