Comparison of pregnenolone sulfate,pregnanolone and estradiol levels between patients with menstrually-related migraine and controls: an exploratory study

BackgroundNeurosteroids affect the balance between neuroexcitation and neuroinhibition but have been little studied in migraine. We compared the serum levels of pregnenolone sulfate, pregnanolone and estradiol in women with menstrually-related migraine and controls and analysed if a correlation existed between the levels of the three hormones and history of migraine and age.MethodsThirty women (mean age ± SD: 33.5 ± 7.1) with menstrually-related migraine (MM group) and 30 aged- matched controls (mean age ± SD: 30.9 ± 7.9) participated in the exploratory study. Pregnenolone sulfate and pregnanolone serum levels were analysed by liquid chromatography-tandem mass spectrometry, while estradiol levels by enzyme-linked immunosorbent assay.ResultsSerum levels of pregnenolone sulfate and pregnanolone were significantly lower in the MM group than in controls (pregnenolone sulfate: P = 0.0328; pregnanolone: P = 0.0271, Student’s t-test), while estradiol levels were similar. In MM group, pregnenolone sulfate serum levels were negatively correlated with history of migraine (R² = 0.1369; P = 0.0482) and age (R² = 0.2826, P = 0.0025) while pregnenolone sulfate levels were not age-related in the control group (R² = 0.04436, P = 0.4337, linear regression analysis).ConclusionLow levels of both pregnanolone, a positive allosteric modulator of the GABAA receptor, and pregnenolone sulfate, a positive allosteric modulator of the NMDA receptor, involved in memory and learning, could contribute either to headache pain or the cognitive dysfunctions reported in migraine patients. Overall, our results agree with the hypothesis that migraine is a disorder associated with a loss of neurohormonal integrity, thus supporting the therapeutic potential of restoring low neurosteroid levels in migraine treatment.Supplementary InformationThe online version contains supplementary material available at 10.1186/s10194-021-01231-9.     

Ischemic necrosis of the femoral head: An experimental rabbit model

To describe the morphology of the proximal femoral epiphysis in a rabbit model from the ischemic insult to the end of the revascularization process. Ischemia of the femoral head was induced in 32 rabbits at the 8th day of life, by sectioning the joint capsule and the ligamentum teres and dislocating the femoral head. Rabbits were sacrificed at 4, 8, 12, 18, 21, 26, 34, and 48 days after surgery and femoral heads were observed histologically. During the first days following the ischemic injury, large areas underwent necrotic changes. Both epiphyseal and physeal cartilage were thicker than normal and less trabecular bone formation was evident. Bone marrow was also diffusely necrotic within the secondary center of ossification. After day 12th, reparative process started with formation of extensive areas of fibrocartilage and several secondary centers of ossifications. At that stage femoral head deformity was already evident. In the following days the secondary centers of ossification cohalesced and epiphyseal and physeal cartilage resumed a normal appearance, but the femur showed a permanent deformity. In newborn rabbits, the ischemic injury to the femoral head blocked the ossification of the epiphyseal and physeal cartilage associated to necrotic bone marrow within the secondary center of ossification of the femoral head as well as to extensive areas of necrosis of epiphyseal and physeal cartilage. Extensive areas of fibrocartilage and small newly formed ossification centers within the femoral epiphysis were the results of the revascularization process, and femoral head deformity became stable afterward. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:535–541, 2015.     

Effects of Azithromycin,Metronidazole, Amoxicillin,and Metronidazole plus Amoxicillin on an In Vitro Polymicrobial Subgingival Biofilm Model

Chronic periodontitis is one of the most prevalent human diseases and is caused by dysbiosis of the subgingival microbiota. Treatment involves primarily mechanical disruption of subgingival biofilms and, in certain cases, adjunctive use of systemic antibiotic therapy. In vitro biofilm models have been developed to study antimicrobial agents targeting subgingival species. However, these models accommodate a limited number of taxa, lack reproducibility, and have low throughput. We aimed to develop an in vitro multispecies biofilm model that mimics subgingival plaque, to test antimicrobial agents. Biofilms were cultivated using the Calgary Biofilm Device and were exposed to amoxicillin (AMX), metronidazole (MTZ), azithromycin (AZM), and AMX-MTZ at four different concentrations for 12, 24, or 36 h. Chlorhexidine (CHX) (0.12%) was used as the positive control. The compositions of the biofilms were analyzed by checkerboard DNA-DNA hybridization, and the percent reduction in biofilm metabolic activity was determined using 2,3,5-triphenyltetrazolium chloride and spectrophotometry. Thirty-five of the 40 species used in the inoculum were consistently recovered from the resulting in vitro biofilms. After 36 h of exposure at the 1:27 dilution, AMX-MTZ reduced metabolic activity 11% less than CHX (q = 0.0207) but 54% more than AMX (q = 0.0031), 72% more than MTZ (q = 0.0031), and 67% more than AZM (q = 0.0008). Preliminary evidence of a synergistic interaction between AMX and MTZ was also observed. In summary, we developed reproducible biofilms with 35 subgingival bacterial species, and our results suggested that the combination of AMX and MTZ had greater antimicrobial effects on these in vitro multispecies biofilms than expected on the basis of the independent effects of the drugs.     

Immunophenotypic features by multiparameter flow cytometry can help distinguish low grade B‐cell lymphomas with plasmacytic differentiation from plasma cell proliferative disorders with an unrelated clonal B‐cell process

Highly sensitive flow cytometry studies may incidentally identify B cell clones when used to assess plasma cell clonality in bone marrows. Clinical history, which can help differentiate related clones (low grade B cell lymphoma with plasmacytic differentiation/LBCL‐PD) from unrelated ones (plasma cell proliferative disorder (PCPD) with an unrelated B cell clone), is often unavailable in referred specimens. We sought to identify morphologic or phenotypic features that would help predict the significance of these clones in the absence of history. We included only cases with identical light chain B and plasma cell clones, as determined by 6‐color flow cytometry with additional DNA ploidy analysis, in which the relationship between clones could be established by review of medical records. There were 26 cases; 18 were related (14 were Waldenstrom macroglobulinemia) and eight were unrelated (seven multiple myeloma). Features seen exclusively in LBCL‐PD include CD19+/CD45+ clonal plasma cell phenotype (66·7%, P = 0·0022) and morphologic features such as paratrabecular bone marrow involvement, increased mast cells, and plasma cells surrounding B‐cell nodules. Aneuploidy was identified exclusively in PCPD cases (75%, P = 0·000028). We conclude that CD19+/CD45+ clonal plasma cell phenotype and aneuploidy are useful in distinguishing related clones (LBCL‐PD) from unrelated clones (PCPD).     

Immune reconstitution syndrome in patients treated for HIV and tuberculosis in Rio de Janeiro.

We made a retrospective longitudinal study from January 2000 to January 2003 to examine cases of immune reconstitution syndrome (IRS) and its incidence rate in tuberculosis (TB)-human immunodeficiency virus (HIV) co-infected patients. The incidence rate (IR) was calculated using a Poisson regression. The confidence interval (CI) that was stipulated was 95%. IRS occurred in 10/84 HIV and TB-positive patients; nine of them were on highly active anti-retroviral therapy (HAART) during a mean of 61.7 (+/- 59) days following the introduction of antiretrovirals. Lymph-node enlargement was the sole clinical manifestation. CD4 counts were <100 cells/mm(3)in 50% of the patients, at the time of TB diagnosis. All but two patients were treated with prednisone, and recovered from TB within a mean of 91 days (+/- 30 days). One relapse of TB was observed, but there were no IRS-related deaths. The incidence rate was higher (IR=11.18; CI, 1.41-88.76) in patients that had superficial lymph node enlargement at the moment of TB diagnosis (not associated with TB), extrapulmonary TB (IR=1.97; CI, 0.44-8.79), were antiretroviral naive (IR=1.85; CI, 0.48-7.16), and CD4 counts <100 cells/mm(3) (IR=1.50; CI, 0.40-5.59), although with a wide CI. IRS was frequent in our sample, occurred more frequently in HIV-naive patients with lymph-node enlargement and extrapulmonary TB. No cases of new pulmonary lesions or worsening of pulmonary infiltrates were observed.     

Oxysterols and EBI2 promote osteoclast precursor migration to bone surfaces and regulate bone mass homeostasis

Bone surfaces attract hematopoietic and nonhematopoietic cells, such as osteoclasts (OCs) and osteoblasts (OBs), and are targeted by bone metastatic cancers. However, the mechanisms guiding cells toward bone surfaces are essentially unknown. Here, we show that the Gαi protein–coupled receptor (GPCR) EBI2 is expressed in mouse monocyte/OC precursors (OCPs) and its oxysterol ligand 7α,25-dihydroxycholesterol (7α,25-OHC) is secreted abundantly by OBs. Using in vitro time-lapse microscopy and intravital two-photon microscopy, we show that EBI2 enhances the development of large OCs by promoting OCP motility, thus facilitating cell–cell interactions and fusion in vitro and in vivo. EBI2 is also necessary and sufficient for guiding OCPs toward bone surfaces. Interestingly, OCPs also secrete 7α,25-OHC, which promotes autocrine EBI2 signaling and reduces OCP migration toward bone surfaces in vivo. Defective EBI2 signaling led to increased bone mass in male mice and protected female mice from age- and estrogen deficiency–induced osteoporosis. This study identifies a novel pathway involved in OCP homing to the bone surface that may have significant therapeutic potential.Osteoclasts (OCs) are multinucleated cells that regulate skeletal development and integrity by actively resorbing excess or damaged bone produced by osteoblasts (OBs) and osteocytes. OBs and osteocytes differentiate from rare mesenchymal stem cells that reside in BM parenchyma (Méndez-Ferrer et al., 2010). In contrast, OCs differentiate from BM-resident and circulatory monocytic precursors that come into close contact with bone surfaces where the essential cytokines ligand for receptor activator of nuclear factor kappa binding (RANKL, encoded by Tnfsf11) and M-CSF (encoded by Csf1) are locally produced by OBs and osteocytes (Teitelbaum, 2000; Nakashima et al., 2011). Cellular interactions between OCs and OBs regulate the activity of both cell types, such that resorbed bone is accurately replaced by newly formed bone. Although the understanding of the molecular signals required for OC differentiation has increased significantly over the past several decades, very little is understood about the mechanisms controlling the migration and positioning of OC precursors (OCPs) near, or in contact with, the bone surface.Monocytes and OCPs are dynamic within BM parenchyma (Ishii et al., 2009). OCPs recirculate between BM and peripheral organs via the action of sphingosine 1-phosphate receptors (S1PRs), which attract multiple hematopoietic cell subsets from BM parenchyma into blood circulation (Walzer et al., 2007; Ishii et al., 2009, 2010; Pereira et al., 2010b). In contrast, the signals promoting cell movement toward bone surfaces remain unknown. However, systemic RANKL administration has been shown to increase OCP homing back to BM and to promote local OC differentiation (Kotani et al., 2013). These studies suggest that OCP movement in and out of BM tissue is highly regulated and that balanced responsiveness to various chemoattractants regulates OCP movement and differentiation. Consistent with this hypothesis, bone resorption produces a milieu that contains potent chemoattractants for monocytes/OCPs (Mundy et al., 1978).Here, we investigated the role played by EBI2, a Gαi protein–coupled receptor (GPCR) involved in dendritic cell and B lymphocyte migration in secondary lymphoid organs (Gatto et al., 2009, 2013; Pereira et al., 2009b; Hannedouche et al., 2011; Kelly et al., 2011; Liu et al., 2011; Yi and Cyster, 2013), in controlling monocyte and OCP movement and positioning within BM. We show that EBI2 is highly expressed in OCPs and mature OCs and promotes OCP motility in vitro and in vivo. Furthermore, we show that OCPs deficient in EBI2 migrate poorly toward bone surfaces, which reduces OC differentiation. In contrast, OCPs that overexpressed EBI2 preferentially localized at the bone surface and fused with preexisting OCs more efficiently than control OCPs. OBs expressed the enzymes CH25H and CYP7B1 that are required for the synthesis of the EBI2-ligand 7α,25-dihydroxycholesterol (7α,25-OHC) and secreted EBI2 ligands in vitro. Interestingly, OCPs also secreted 7α,25-OHC, which results in autocrine EBI2 signaling and tempers EBI2-mediated migration toward bone surfaces. Finally, EBI2 signaling–deficient mice exhibit increased bone mass at young and old ages, and EBI2 signaling deficiency significantly protects female mice from osteoporosis induced by estrogen deficiency.     

Impact of the COVID-19 Pandemic on Health-Related Concerns,Quality of Life and Psychological Adjustment in Young Adults with Congenital Heart Disease

Background: The risk for a severe disease course in case of infection with SARS-CoV-2 in young adults with congenital heart disease is largely unknown, potentially leading to uncertainty and anxiety among affected patients. This study aims to investigate health-related concerns, health-related quality of life and psychological adjustment in patients with congenital heart disease compared to healthy peers during the COVID-19 pandemic. Methods: One-hundred patients with congenital heart disease and 50 controls (M = 29.7, SD = 3.8 years) were recruited. They completed an online survey including the assessment of health-related concerns regarding COVID-19, the 12-item Short Form Health Survey and the Brief Symptom Inventory. Results: Patients considered COVID-19 to be a more serious issue (Generalized odds ratio [GenOR] = 1.67, p = 0.04), were more concerned about becoming infected (GenOR = 2.93, p < 0.001) and expressed more fear about leaving their homes (GenOR = 1.81, p = 0.004) while general anxiety symptoms were not different between groups (p = 0.23). Patients relied more on family and friends for support (30% vs. 2% in controls, p < 0.001) and reported better compliance with protective measures (p = 0.03). Mental health-related quality of life and psychological adjustment were not different between groups (p = 0.17 and p = 0.68, respectively). Physical health-related quality of life was lower in patients compared to controls (p = 0.03). Conclusions: Young adults with congenital heart disease in Switzerland are more concerned about their health during the COVID-19 pandemic compared to healthy peers. These concerns, however, do not translate into generally impaired mental wellbeing. The impact of the easing of lockdown measures on long-term anxiety levels and quality of life requires further study