Cell cycle and mitosis progression during ZIKA virus infection: The viral non-structural protein NS5 as a master regulator of the APC/cyclosome?

Alterations in cell cycle regulation contribute to Zika virus (ZIKV)-associated pathogenesis and may have implications for the development of therapeutic avenues. As a matter of fact, ZIKV alters cell cycle progression at multiple stages, including G1, S, G2, and M phases. During a cell cycle, the progression of mitosis is particularly controlled to avoid any abnormalities in cell division. In this regard, the critical metaphase-anaphase transition is triggered by the activation of anaphase-promoting complex/cyclosome (APC/C) by its E3 ubiquitin ligase subunit Cdc20. Cdc20 recognizes substrates by interacting with a destruction box motif (D-box). Recently, the ZIKV nonstructural protein 5 (NS5), one of the most highly conserved flavivirus proteins, has been shown to localize to the centrosome in each pole and to spindle fibers during mitosis. Inducible expression of NS5 reveals an interaction of this viral factor with centrosomal proteins leading to an increase in the time required to complete mitosis. By analyzing the NS5 sequence, we discovered the presence of a D-box. Taken together, these data support the idea that, in addition to its role in viral replication, NS5 plays a critical role in the control of the cell cycle of infected cells and, more specifically, in the regulation of the mitotic spindle. Here we propose that the NS5 protein may interfere with the metaphase-anaphase progression, and thus cause the observed delay in mitosis via the regulation of APC/C.

Outcome of reverse shoulder arthroplasty secondary to rotator cuff arthropathy in a low-income population.

BACKGROUND: Reverse shoulder arthroplasty (RSA) is a valuable treatment for rotator cuff arthropathy (RCA) in developed regions. Socioeconomic issues impact access to specialized care and there is a lack of data on RSA outcomes in developing regions. We present our 24-month follow-up on RSA surgeries to treat RCA in our low-income population. METHODS: Prospective evaluation of 26 patients subjected to RSA at Hospital Geral de Fortaleza-CE, Brazil, between January 2018 and December 2020. Literacy [>/≤ 8 school years(SY)] and income were documented. Outcomes considered pain (visual analogue scale; VAS) as well as SSV, SPADI, ASES, and UCLA scoring, and range of motion [forward flexion (FF); external rotation (ER)]. RESULTS: Patients were 68.5 ± 7.6 years-old with 16(61.5%) females; 65% had hypertension and 7 (26.9%) had diabetes. Over 90% declared < 900.00 US$ monthly family earnings and 10 (38.4%) patients declared ≤8 SY with > 80% exerting blue-collar jobs. Pain showed a significant reduction from baseline (8 ± 2) to 24 months (2.1 ± 2.3; p < 0.001). UCLA (10.3 ± 5.6 and 28.6 ± 7.2), ASES (16.7 ± 10.8 and 63.1 ± 28.4), SSV (326 ± 311 and 760 ± 234), and SPADI (98.3 ± 26.5) scores significantly improved from baseline to 24 months, achieving minimal clinically important difference. FF (89.2° ± 51.2° to 140.6 ± 38.3°) and ER (19.2° ± 22.5 to 33.4° ± 20.6°) significantly improved from baseline to 24 months (p = 0.004 and 0.027, respectively). There were 5 non-serious adverse events with one surgical revision. All patients returned to daily life activities. CONCLUSION: This is the first outcome report 2 years following RSA in a low-income population. Data indicate this procedure is justifiable regardless of socioeconomic issues.

Comparison of accuracy and reliability of CBCT and 3D laser scanner in the volumetric assessment of the root canal space.

PURPOSE: To compare the accuracy and reliability of cone-beam computed tomography (CBCT) and laser scanner in measuring minor volume changes such as the root canal space. METHODS: 35 maxillary incisors were endodontically prepared. A dimensionally stable silicone material was injected into the root canal space and scanned with CBCT. The root canal volume was measured using Romexis 3.0.1 R software. Replicas were carefully removed from the teeth and scanned using an extraoral laser scanner. These images were exported to the Rhinoceros software for volume measurement. The volume of each replica was also assessed using the gravimetric method. To determine the accuracy, the volume obtained from both devices was compared with the gravimetric method. Statistical analysis was done using a paired t-test. The reliability was assessed using the intraclass correlation coefficient. RESULTS: There was no statistically significant difference between the mean volume of CBCT 27.04 ± 7.25 mm³ and the mean volume of the gravimetric method 27.87 ± 7.17 mm³ (P< 0.05). A statistically significant difference was seen with the laser scanner at 25.31 ± 6.89 mm³ and the gravimetric method at 27.87 ± 7.17 mm³ (P< 0.05). CBCT showed a good degree of agreement (ICC 0.899), while the laser scanner showed a moderate degree of agreement (ICC 0.644) with the gravimetric method. CBCT proved accurate and reliable in measuring minor volumes like the root canal space, ideally in the range of 20-25 mm³. The laser scanner presented acceptable reliability. CLINICAL SIGNIFICANCE: The laboratory data showed satisfactory outcomes, providing an evidence-based approach and potentially motivating clinicians to integrate cone-beam computed tomography for volume analysis into clinical practice. The accuracy and reliability of laser scanners for small-volume analysis have not previously been evaluated. Consequently, the findings from this study warrant further clinical investigations.

Fermented Pineapple Juice Consumption Limits Metabolic Disorders Associated to Sugary Drinks on High-Fat Diet-Fed Mice.

SCOPE: Lactic acid fermentation (LAF) modulates the composition of food, leading to changes in safety, sensory, and nutritional properties. The effects of lactic fermented pineapple juice (FJ) supplementation on energetic metabolism of high-fat diet (HFD) fed mice are compared with either water (control), sweetened water (SW), bacteria in SW, and pineapple juice (J) supplementation. METHODS AND RESULTS: Drink consumption and body weight are measured during the 6 weeks of experiment, whereas glycemia and lipid content are determined at the beginning and at the end of the experiment. Total energy intake is similar between all groups though the volume of juice consumed is lower than that of SWs. Weight gain is higher for mice provided with sugary drinks (5.65 ± 1.32 to 7.74 ± 2.98 g) compared to water (4.68 ± 0.93 g). The FJ is less detrimental to blood carbohydrate regulation than other sugary drinks. Triglyceride (TG) and total cholesterol content are not modified following fermented juice or water consumption, contrarily to other sugary drinks. Whatever the drink, intestinal permeability is preserved. Lactic acid bacterium (LAB) population in feces is not affected by the beverage but species composition is modified. CONCLUSION: From a health perspective, FJ is preferable to other sugary drinks to limit metabolic disorders related to HFD.

Improving connections to early childhood systems of care via a universal home visiting program in Massachusetts.

Welcome Family is a universal, short-term nurse home visiting program designed to promote optimal maternal and infant physical and mental well-being and provide an entry point into the early childhood system of care to all families with newborns up to 8 weeks old living in defined communities in Massachusetts. The present study examines whether: 1) Welcome Family meets its goal of successfully connecting families to two early childhood programs-evidence-based home visiting (EBHV) and early intervention (EI)-relative to families with similar background experiences who do not participate in Welcome Family, and 2) whether these impacts are conditional on families' race and ethnicity and their primary language-two characteristics that are related to structural racism and health inequities. The study used coarsened exact matching (CEM) based on birth certificate data to match Welcome Family participants who enrolled during 2013-2017 to mothers and their infants living in the home visiting catchment areas who did not receive home visiting during the study period. Primary study outcomes included enrollment in any EBHV program supported by the Massachusetts Maternal, Infant, and Early Childhood Home Visiting (MA MIECHV) program up to age 1 year, measured using MA MIECHV home visiting program data, and EI service receipt for children aged up to age 3 years, measured using EI program data. Impacts were assessed by fitting weighted regression models adjusted for preterm birth, maternal depression, and substance use. Mothers' race, ethnicity, and language were included in the model as moderators of Welcome Family impacts on enrollment in EBHV and EI. Welcome Family participants (n = 3,866) had more than double the odds of EBHV enrollments up to age 1 and had 1.39 greater odds of receiving EI individualized family service plans (IFSPs) up to age 3 relative to the comparison group (n = 46,561). Mothers' primary language moderated Welcome Family impacts on EBHV enrollments. Universal, short-term programs such as Welcome Family may be an effective method of ensuring families who could benefit from more intensive early childhood services are identified, engaged, and enrolled.

The Massachusetts Racial Equity Data Road Map: Data as a Tool Toward Ending Structural Racism.

BACKGROUND: In 2015, the Massachusetts Department of Public Health (MDPH) adopted a Title V maternal and child health priority to "promote health and racial equity by addressing racial justice and reducing disparities." A survey assessing staff capacity to support this priority identified data collection and use as opportunities for improvement. In response, MDPH initiated a quality improvement project to improve use of data for action to promote racial equity. METHODS: MDPH conducted value stream mapping to understand existing processes for using data to inform racial equity work. Key informant interviews and a survey of program directors identified challenges to using data to promote racial equity. MDPH used a cause-and-effect diagram to identify and organize challenges to using data to inform racial equity work and better understand opportunities for improvement and potential solutions. RESULTS: Key informants highlighted the need to consider structural factors and historical and community contexts when interpreting data. Program directors noted limited staff time, lack of performance metrics, competing priorities, low data quality, and unclear expectations as challenges. To address the identified challenges, the team identified potential solutions and prioritized development and piloting of the MDPH Racial Equity Data Road Map (Road Map). CONCLUSIONS: The Road Map framework provides strategies for data collection and use that support the direction of actionable data-driven resources to racial inequities. The Road Map is a resource to support programs to authentically engage communities; frame data in the broader contexts that impact health; and design solutions that address root causes. With this starting point, public health systems can work toward creating data-driven programs and policies to improve racial equity.

Surgical outcome following rotator cuff tear repair in a low-income population. Impact of obesity and smoking.

BACKGROUND: Comorbidities and socioeconomic issues impact outcome of rotator cuff tear (RCT) repair. There are no data on RCT repair outcome from developing regions. We determined the impact of obesity and smoking following RCT repair in a low-income population. METHODS: This is a retrospective case series. Forty-seven shoulders of 42 patients subjected to open or arthroscopic repair of a RCT with a minimum of 2 years follow-up were cross-sectionally evaluated. Patients were seen in the Orthopaedic Service of the Hospital Geral de Fortaleza-CE, Brazil between March and September 2018. RCT were classified as partial or full-thickness lesions. Fatty infiltration (Goutallier) and tendon retraction (Patte) were recorded as well as obesity (BMI > 30), literacy [>/≤ 8 school years (SY)] and smoking status 6 months prior to surgery (present/absent). Outcomes included pain (visual analogue scale; VAS, 0-10 cm), range of motion [active forward flexion and external rotation (ER)], UCLA and ASES scoring. RESULTS: Patients were 59.9 ± 7.4 years-old, 35(74.4%) female with 19 (17.1-30.2 IQR) median of months from diagnosis to surgery and 25 median months of follow-up (26.9-34.0 IQR); over 90% declared < 900.00 US$ monthly family income and two-thirds had ≤8 SY. Forty patients (85.1%) had full-thickness tears, 7 (14.9%) had Goutallier ≥3 and over 80% had < Patte III stage. Outcomes were similar regardless of fatty infiltration or tendon retraction staging. There were 17 (36.1%) smokers and 13 (27.6%) obese patients. Outcome was similar when comparing obese vs non-obese patients. Smokers had more pain (P = 0.043) and less ER (P = 0.029) with a trend towards worse UCLA and ASES scores as compared to non-smokers though differences did not achieve minimal clinically important difference (MCID) proposed for surgical RCT treatment. After adjusting for obesity, VAS and ER values in smokers were no longer significant (P = 0.2474 and 0.4872, respectively). CONCLUSIONS: Our data document outcomes following RCT repair in a low-income population. Smoking status but not obesity impacted RCT repair outcome though not reaching MCID for surgical treatment.

Protective Effects of Medicinal Plant Decoctions on Macrophages in the Context of Atherosclerosis.

Atherosclerosis is a hallmark of most cardiovascular diseases. The implication of macrophages in this pathology is widely documented, notably for their contribution to lipid accumulation within the arterial wall, associated with oxidative stress and inflammation processes. In order to prevent or limit the atherosclerosis damage, nutritional approaches and medicinal plant-based therapies need to be considered. In Reunion Island, medicinal plant-based beverages are traditionally used for their antioxidant, lipid-lowering and anti-inflammatory properties. The aim of our study was to assess the protective effects of eight medicinal plant decoctions in an in vitro model of RAW 264.7 murine macrophages exposed to pro-atherogenic conditions (oxidized low-density lipoproteins-ox-LDL-E. coli Lipopolysaccharides-LPS). The impact of polyphenol-rich medicinal plant decoctions on cell viability was evaluated by Neutral Red assay. Fluorescent ox-LDL uptake was assessed by flow cytometry and confocal microscopy. Activation of NF-κB was evaluated by quantification of secreted alkaline phosphatase in RAW-Blue™ macrophages. Our results show that medicinal plant decoctions limited the cytotoxicity induced by ox-LDL on macrophages. Flow cytometry analysis in macrophages demonstrated that medicinal plant decoctions from S. cumini and P. mauritianum decreased ox-LDL uptake and accumulation by more than 70%. In addition, medicinal plant decoctions also inhibited NF-κB pathway activation in the presence of pro-inflammatory concentrations of E. coli LPS. Our data suggest that medicinal plant decoctions exert protective effects on ox-LDL-induced cytotoxicity and limited macrophage lipid uptake. Moreover, herbal preparations displayed anti-inflammatory properties on macrophages that can be of interest for limiting the atherosclerotic process.

Neurothekeoma in the Axilla Causing Persistent Shoulder Pain: Case Report.

Neurothekeomas, also known as neural sheath myxomas, are rare benign tumors of the neural sheath affecting most commonly the head, arms and shoulder of women in their 2 nd and 3 rd decades of life. Due to the low prevalence and undefined clinical picture, they are hardly considered in the initial differential diagnosis of skin tumors. We report the case of a 24 year-old woman who was seen in 2016 reporting > 1 year of moderate pain and limited mobility of her left shoulder. Clinical evaluation revealed restricted mobility of the affected shoulder and nuclear magnetic resonance imaging showed a T2-weighted contrast-enhanced multilobular mass in the quadrilateral area apparently invading the adjacent humeral cortical region. Histopathology of a needle sample material revealed loose fibroconnective tissue with no signs of invasion, mitosis or atypical figures. Successful surgical excision was performed and the diagnosis of neurothekeoma was confirmed after detailed histopathology, including immunohistochemistry. The patient was asymptomatic at 18 months of follow-up, with full recovery of shoulder movement and no signs of relapse.

Neurothekeoma in the Axilla Causing Persistent Shoulder Pain: Case Report / Neurotecoma na axila causando dor persistente no ombro: Relato de caso

Abstract Neurothekeomas, also known as neural sheath myxomas, are rare benign tumors of the neural sheath affecting most commonly the head, arms and shoulder of women in their 2nd and 3rd decades of life. Due to the low prevalence and undefined clinical picture, they are hardly considered in the initial differential diagnosis of skin tumors. We report the case of a 24 year-old woman who was seen in 2016 reporting > 1 year of moderate pain and limited mobility of her left shoulder. Clinical evaluation revealed restricted mobility of the affected shoulder and nuclear magnetic resonance imaging showed a T2-weighted contrast-enhanced multilobular mass in the quadrilateral area apparently invading the adjacent humeral cortical region. Histopathology of a needle sample material revealed loose fibroconnective tissue with no signs of invasion, mitosis or atypical figures. Successful surgical excision was performed and the diagnosis of neurothekeoma was confirmed after detailed histopathology, including immunohistochemistry. The patient was asymptomatic at 18 months of follow-up, with full recovery of shoulder movement and no signs of relapse.

Resumo Neurotecomas, também conhecidos como mixomas da bainha neural, são tumores benignos raros da bainha neural afetando mais comumente a cabeça, braços e ombros de mulheres entre 20 e 40 anos de idade. Devido à baixa prevalência e quadro clínico mal definido, essas lesões são raramente consideradas no diagnóstico diferencial de tumores cutâneos. Relatamos o caso de uma mulher de 24 anos de idade que procurou atendimento em 2016 relatando dor moderada por mais de um ano e limitação dos movimentos do ombro esquerdo. Ao exame, foi constatada restrição da mobilidade dessa articulação e uma ressonância magnética revelou imagem multilobular com aumento de sinal em T2 na região quadrilateral, aparentando invasão da região cortical do úmero subjacente. A histopatologia de uma biópsia incisional mostrou lesão composta por tecido conjuntivo frouxo, sem sinais de invasão, figuras de mitose ou atipias. Foi realizada excisão completa da lesão e o diagnóstico de neurotecoma foi confirmado após análise histopatológica que incluiu painel imunohistoquímico. À revisão de 18 meses, a paciente estava assintomática com recuperação completa do movimento e sem evidência de recidiva da lesão.

Evaluation of Polyphenol Content and Antioxidant Capacity of Aqueous Extracts from Eight Medicinal Plants from Reunion Island: Protection against Oxidative Stress in Red Blood Cells and Preadipocytes.

Background-Medicinal plants are traditionally used as infusions or decoctions for their antioxidant, anti-inflammatory, hypolipidemic and anti-diabetic properties. Purpose-The aim of the study was to define the polyphenol composition and to assess the antioxidant capacity of eight medicinal plants from Reunion Island referred to in the French Pharmacopeia, namely Aphloia theiformis, Ayapana triplinervis, Dodonaea viscosa, Hubertia ambavilla, Hypericum lanceolatum, Pelargonium x graveolens, Psiloxylon mauritianum and Syzygium cumini. Methods-Polyphenol content was assessed by biochemical assay and liquid chromatography coupled to mass spectrometry. Antioxidant capacity was assessed by measuring DPPH reduction and studying the protective effects of herbal preparation on red blood cells or preadipocytes exposed to oxidative stress. Results-Polyphenol content ranged from 25 to 143 mg gallic acid equivalent (GAE)/L for infusions and 35 to 205 mg GAE/L for decoctions. Liquid chromatography coupled to mass spectrometry analysis showed the presence of major bioactive polyphenols, such as quercetin, chlorogenic acid, procyanidin and mangiferin. Antioxidant capacity assessed by different tests, including DPPH and Human red blood cell (RBC) hemolysis of herbal preparations, demonstrated a dose-dependent effect whatever the extraction procedure. Our data suggest that decoction slightly improved polyphenol extraction as well as antioxidant capacity relative to the infusion mode of extraction (DPPH test). However, infusions displayed a better protective effect against oxidative stress-induced RBC hemolysis. Conclusion-Traditional preparations of medicinal plant aqueous extracts (infusions and decoctions) display antioxidant properties that limit oxidative stress in preadipocytes and red blood cells, supporting their use in the context of metabolic disease prevention and treatment.

Effects of exposure to large sharks on the abundance and behavior of mobile prey fishes along a temperate coastal gradient.

Top predators can exert strong influences on community structure and function, both via direct, consumptive effects, as well as through non-consumptive, fear-based effects (i.e. predation risk). However, these effects are challenging to quantify, particularly for mobile predators in marine ecosystems. To advance this field of research, here we used baited remote underwater video stations (BRUVs) to assess how the behavior of mobile fish species off Cape Cod, Massachusetts, was affected by exposure to large sharks. We categorized sites into three levels of differential shark predation exposure (white sharks, Carcharodon carcharias) and quantified the relative abundance and arrival times (elapsed time before appearing on screen) for six mobile fish prey groups to the BRUV stations. Increased large shark exposure was associated with a decrease in overall prey abundance, but the overall response was prey group-specific. Foraging of smooth dogfish, a likely important prey item for large sharks in the system, was significantly reduced in areas frequented by white sharks. Specifically, the predicted probabilities of smooth dogfish bait contacts or bite attempts occurring were reduced by factors of 5.7 and 8.4, respectively, in areas of high exposure as compared to low exposure. These modifications were underscored by a decrease in smooth dogfish abundance in areas of high exposure as well. Our results suggest that populations of large, roving sharks may induce food-related costs in prey. We discuss the implications of this work within the context of the control of risk (COR) hypothesis, for the purposes of advancing our understanding of the ecological role and effects of large sharks on coastal marine ecosystems.

Participant characteristics in autism intervention studies.

The purpose of this Letter to the Editor is to discuss the recent paper, "Lessons learned: Engaging culturally diverse families in neurodevelopmental disorders intervention research" by Ratto et al. Specifically, we are interested in further exploring the question of "who participates in autism spectrum disorder intervention research," and how this question may impact interpretation of Ratto and colleagues' paper.

Lessons Learned: Implementation of Pilot Universal Postpartum Nurse Home Visiting Program, Massachusetts 2013-2016.

Purpose Home visiting programs for new families in the United States have traditionally served high-risk families. In contrast, universal home visiting models serve all families regardless of income, age, risk or other criteria. They offer an entry point into a system of care for children and families, with the potential to improve population health. This paper describes lessons learned from the first three years of implementing a universal home visiting model. Description Welcome Family is a universal home visiting program in Massachusetts that offers a one-time visit by a nurse to new mothers up to eight weeks postpartum. The Massachusetts Department of Public Health (MDPH) is piloting Welcome Family in four communities with the goal of expanding statewide. Assessment Welcome Family served over 3000 families in its first three years. Program performance measures provided a framework to examine successes and challenges related to outreach and enrollment, program operations, and linkages with community resources. Early challenges included increasing referrals to a new program and limited capacity to serve all women giving birth. Local implementing agencies tested innovative strategies and MDPH made program modifications, such as developing quarterly data reports and establishing a learning collaborative, to address identified challenges. Conclusion MDPH is committed to the success of Welcome Family and uses continuous quality improvement to maximize the impact of the program on families and the system of care in Massachusetts. Lessons learned from the Massachusetts pilot can inform other states' efforts to enhance their early childhood systems of care through expanding universal home visiting.

Porphyromonas gingivalis lipopolysaccharides act exclusively through TLR4 with a resilience between mouse and human.

Porphyromonas gingivalis is a key bacterium in chronic periodontitis, which is associated with several chronic inflammatory diseases. Lipopolysaccharides from P. gingivalis (Pg LPS) can activate multiple cell types via the production of pro-inflammatory cytokines. The receptors for Pg LPS have initially been reported as TLR2, contrasting with the well-studied TLR4 receptor for E. coli LPS; this observation remains controversial since synthetic Pg lipid A activates TLR4 but not TLR2. Despite this observation, the dogma of Pg LPS-mediated TLR2 activation remains the basis of many hypotheses and result interpretations. In the present work, we aimed at determining whether TLR4 or TLR2, or both, mediate Pg LPS pro-inflammatory activity using Pg LPS with different grades of purity, instead of synthetic lipid A from Pg LPS. Here we show that Pg LPS 1) acts exclusively through TLR4, and 2) are differently recognized by mouse and human TLR4 both in vitro and in vivo. Taken together, our results suggest that Pg LPS activity is mediated exclusively through TLR4 and only weakly induces proinflammatory cytokine secretion in mouse models. Caution should be taken when extrapolating data from mouse systems exposed to Pg or Pg LPS to humans.

Structural characteristics are crucial to the benefits of guar gum in experimental osteoarthritis.

Protein-free guar gum (DGG) was oxidized (DGGOX) or sulfated (DGGSU) by insertion of new groups in C-6 (manose) and C-6 (galactose), for DGGOX and DGGSU, respectively. Rats were subjected to anterior cruciate ligament transection (ACLT) of the knee, joint pain recorded using the articular incapacitation test, and the analgesic effect of intraarticular 100µg DGG, DGGOX or DGGSU solutions at days 4-7 was evaluated. Other groups received DGG or saline weekly, from days 7 to 70 and joint damage assessed using histology and biochemistry as the chondroitin sulfate (CS) content of cartilage. The molar mass of CS samples was obtained by comparing their relative electrophoretic mobility to standard CS. DGG but not DGGOX or DGGSU significantly inhibited joint pain. DGG significantly reversed the increase in CS, its reduced electrophoretic mobility, and histological changes following ACLT, as compared to vehicle. Structural integrity accounts for DGG benefits in experimental osteoarthritis.

Stent-assisted coil embolization of aneurysms with small parent vessels: safety and efficacy analysis.

BACKGROUND: Stent-assisted coil embolization (SACE) is a viable therapeutic approach for wide-neck intracranial aneurysms. However, it can be technically challenging in small cerebral vessels (≤2 mm). OBJECTIVE: To present our experience with stents approved for SACE in aneurysms with small parent arteries. METHODS: All patients who underwent stent-assisted aneurysm treatment with either a Neuroform or an Enterprise stent device at our institution between June 2006 and October 2012 were identified. Additionally, we evaluated each patient's vascular risk factors, aneurysm characteristics (ruptured vs non-ruptured, incidental finding, recanalized) and follow-up angiography data. RESULTS: A total of 41 patients with 44 aneurysms met our criteria, including 31 women and 10 men. Most of the aneurysms were located in the anterior circulation (75%). Stent placement in vessels 1.2-2 mm in diameter was successful in 93.2%. Thromboembolic complications occurred in 6 cases and vessel straightening was seen in 1 case only. Initial nearly complete to complete aneurysm obliteration was achieved in 88.6%. Six-month follow-up angiography showed coil compaction in three cases, one asymptomatic in-stent stenosis and stent occlusion. Twelve to 20-months' follow-up showed stable coil compaction in two patients compared with previous follow-up, and aneurysm recanalization in two patients. Twenty-four to 36-months' follow-up showed further coil compaction in one of these patients and aneurysm recanalization in a previous case of stable coil compaction on mid-term follow-up. CONCLUSIONS: Our results suggest that SACE of aneurysms with small parent vessels is feasible in selected cases and shows good long-term patency rates of parent arteries.

Recombinant human HSP60 produced in ClearColi™ BL21(DE3) does not activate the NFκB pathway.

HSP60, an intracellular molecular chaperone has been largely described as an alarmin or damage-associated molecular pattern when released outside the cell. HSP60 has been reported as a possible ligand of TLR2 or TLR4 inducing NFκB-dependant signaling pathway leading to cytokine secretion. However, recent publications suggested that HSP60 could not act as an activator of TLR4 by itself. The observed effect could be due to the presence of endotoxin in HSP60 preparation especially LPS. In order to clarify the controversy, we produced recombinant human HSP60 in two different strains of Escherichia coli, standard strain for protein overproduction, BL21(DE3), and the new ClearColi BL21(DE3) strain which lacks LPS-activity through TLR4. Undoubtedly, we have shown that recombinant HSP60 by itself was not able to induce NFκB-dependant signaling pathway in a model of THP1 monocyte cell line. Our data suggest that HSP60 needs either pathogen-associated molecules, specific post-translational modification and/or other host factors to activate immune cells via NFκB activation.

Antioxidant polyphenol-rich extracts from the medicinal plants Antirhea borbonica, Doratoxylon apetalum and Gouania mauritiana protect 3T3-L1 preadipocytes against H2O2, TNFα and LPS inflammatory mediators by regulating the expression of superoxide dismutase and NF-κB genes.

BACKGROUND: Adipose cells responsible for fat storage are the targets of reactive oxygen species (ROS) like H2O2 and pro-inflammatory agents including TNFα and LPS. Such mediators contribute to oxidative stress and alter inflammatory processes in adipose tissue, leading to insulin resistance during obesity. Thus, the identification of natural compounds such as plant polyphenols able to increase the antioxidant and anti-inflammatory capacity of the body is of high interest. We aimed to evaluate the biological properties of polyphenol-rich extracts from the medicinal plants A. borbonica, D. apetalum and G. mauritiana on preadipocytes exposed to H2O2, TNFα or LPS mediators. METHODS: Medicinal plant extracts were analysed for their polyphenol contents by Folin-Ciocalteu and UPLC-ESI-MS methods as well as for their free radical-scavenging activities by DPPH and ORAC assays. To assess the ability of polyphenol-rich extracts to protect 3T3-L1 preadipocytes against H2O2, TNFα or LPS mediators, several parameters including cell viability (MTT and LDH assays), ROS production (DCFH-DA test), IL-6 and MCP-1 secretion (ELISA) were evaluated. Moreover, the expression of superoxide dismutase, catalase and NF-κB genes was explored (RT-QPCR). RESULTS: All medicinal plants exhibited high levels of polyphenols with free radical-scavenging capacities. Flavonoids such as quercetin, kaempferol, epicatechin and procyanidins, and phenolic acids derived from caffeic acid including chlorogenic acid, were detected. Polyphenol-rich plant extracts did not exert a cytotoxic effect on preadipocytes but protected them against H2O2 anti-proliferative action. Importantly, they down-regulated ROS production and the secretion of IL-6 and MCP-1 pro-inflammatory markers induced by H2O2, TNFα and LPS mediators. Such a protective action was associated with an increase in superoxide dismutase antioxidant enzyme gene expression and a decrease in mRNA levels of NF-κB pro-inflammatory transcription factor. CONCLUSION: This study highlights that antioxidant strategies based on polyphenols derived from medicinal plants tested could contribute to regulate adipose tissue redox status and immune process, and thus participate to the improvement of obesity-related oxidative stress and inflammation.

A randomized, controlled trial of corneal collagen cross-linking in progressive keratoconus: three-year results.

PURPOSE: To report the refractive, topographic, and clinical outcomes 3 years after corneal collagen cross-linking (CXL) in eyes with progressive keratoconus. DESIGN: Prospective, randomized controlled trial. PARTICIPANTS: One hundred eyes with progressive keratoconus were randomized into the CXL treatment or control groups. METHODS: Cross-linking was performed by instilling riboflavin 0.1% solution containing 20% dextran for 15 minutes before and during the 30 minutes of ultraviolet A irradiation (3 mW/cm(2)). Follow-up examinations were arranged at 3, 6, 12, 24, and 36 months. MAIN OUTCOME MEASURES: The primary outcome measure was the maximum simulated keratometry value (Kmax). Other outcome measures were uncorrected visual acuity (UCVA; measured in logarithm of the minimum angle of resolution [logMAR] units), best spectacle-corrected visual acuity (BSCVA; measured in logMAR units), sphere and cylinder on subjective refraction, spherical equivalent, minimum simulated keratometry value, corneal thickness at the thinnest point, endothelial cell density, and intraocular pressure. RESULTS: The results from 48 control and 46 treated eyes are reported. In control eyes, Kmax increased by a mean of 1.20±0.28 diopters (D), 1.70±0.36 D, and 1.75±0.38 D at 12, 24, and 36 months, respectively (all P <0.001). In treated eyes, Kmax flattened by -0.72±0.15 D, -0.96±0.16 D, and -1.03±0.19 D at 12, 24, and 36 months, respectively (all P <0.001). The mean change in UCVA in the control group was +0.10±0.04 logMAR (P = 0.034) at 36 months. In the treatment group, both UCVA (-0.15±0.06 logMAR; P = 0.009) and BSCVA (-0.09±0.03 logMAR; P = 0.006) improved at 36 months. There was a significant reduction in corneal thickness measured using computerized videokeratography in both groups at 36 months (control group: -17.01±3.63 µm, P <0.001; treatment group: -19.52±5.06 µm, P <0.001) that was not observed in the treatment group using the manual pachymeter (treatment group: +5.86±4.30 µm, P = 0.181). The manifest cylinder increased by 1.17±0.49 D (P = 0.020) in the control group at 36 months. There were 2 eyes with minor complications that did not affect the final visual acuity. CONCLUSIONS: At 36 months, there was a sustained improvement in Kmax, UCVA, and BSCVA after CXL, whereas eyes in the control group demonstrated further progression.