Blood copper and risk of cardiometabolic diseases: a Mendelian randomization study.

Observational evidence links higher blood levels of copper with higher risk of cardiovascular diseases. However, whether those associations reflect causal links or can be attributed to confounding is still not fully clear. We investigated causal effects of copper on the risk of cardiometabolic endpoints (stroke, coronary artery disease [CAD] and type 2 diabetes) and cardiometabolic risk factors in two-sample Mendelian randomization (MR) studies. The selection of genetic instruments for blood copper levels relied on meta-analysis of genome-wide association studies in three independent studies (European Prospective Investigation into Cancer and Nutrition-Potsdam study, Prospective investigation of the Vasculature in Uppsala Seniors study, Queensland Institute of Medical Research studies). For the selected instruments, outcome associations were drawn from large public genetic consortia on the respective disease endpoints (MEGASTROKE, Cardiogram, DIAGRAM) and cardiometabolic risk factors. MR results indicate an inverse association for genetically higher copper levels with risk of CAD (odds ratio [95% confidence interval] = 0.92 [0.86-0.99], P = 0.022) and systolic blood pressure (beta [standard error (SE)] = -0.238 [0.121]; P = 0.049). Multivariable MR incorporating copper and systolic blood pressure into one model suggested systolic blood pressure as mediating factor between copper and CAD risk. In contrast to previous observational evidence establishing higher blood copper levels as risk-increasing factor for cardiometabolic diseases, this study suggests that higher levels of genetically predicted copper might play a protective role for the development of CAD and systolic blood pressure.

Pain management in the postoperative period of amputation surgeries: A scoping review.

BACKGROUND: Pain in the postoperative period of amputation surgeries, when not managed correctly, can have consequences for the patient. PURPOSES AND OBJECTIVES: The aim of this study was to map the scientific evidence on pain management in patients in the postoperative period of amputation surgeries. DESIGN: Scoping review with elaboration based on the recommendations of the Joanna Briggs Institute, supported by The PAGER framework and guided by the PRISMA-ScR Checklist. METHODS: The survey was conducted in August 2022 in 10 data sources. The 3300 publications found were analysed by two independent reviewers, after applying inclusion and exclusion criteria, 16 studies were selected. RESULTS: Publications occurred between the years 1997 and 2022 and mainly dealt with the management of residual pain and phantom limb through pharmacological measures with pain assessment made by numerical scales. CONCLUSIONS: The literature pointed out strategic uses of analgesics and highlighted technologies for neurostimulation through catheters. The need for new studies with non-pharmacological measures and with designs that can prove their effectiveness is reinforced. RELEVANCE TO CLINICAL PRACTICE: The knowledge of these strategies by health professionals allows adequate pain management and patient follow-up for a less traumatic recovery. PATIENT OR PUBLIC CONTRIBUTION: To patient or public involvement in this scoping review.

A live auxotrophic vaccine confers mucosal immunity and protection against lethal pneumonia caused by Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the leading causes of nosocomial pneumonia and its associated mortality. Moreover, extensively drug-resistant high-risk clones are globally widespread, presenting a major challenge to the healthcare systems. Despite this, no vaccine is available against this high-concerning pathogen. Here we tested immunogenicity and protective efficacy of an experimental live vaccine against P. aeruginosa pneumonia, consisting of an auxotrophic strain which lacks the key enzyme involved in D-glutamate biosynthesis, a structural component of the bacterial cell wall. As the amounts of free D-glutamate in vivo are trace substances in most cases, blockage of the cell wall synthesis occurs, compromising the growth of this strain, but not its immunogenic properties. Indeed, when delivered intranasally, this vaccine stimulated production of systemic and mucosal antibodies, induced effector memory, central memory and IL-17A-producing CD4+ T cells, and recruited neutrophils and mononuclear phagocytes into the airway mucosa. A significant improvement in mice survival after lung infection caused by ExoU-producing PAO1 and PA14 strains was observed. Nearly one third of the mice infected with the XDR high-risk clone ST235 were also protected. These findings highlight the potential of this vaccine for the control of acute pneumonia caused by this bacterial pathogen.

In vivo monitoring of Lactiplantibacillus plantarum in the nasal and vaginal mucosa using infrared fluorescence.

Lactic acid bacteria (LAB) of the genus Lactiplantibacillus have been explored as potential mucosal vaccine vectors due to their ability to elicit an immune response against expressed foreign antigens and to their safety. However, tools for monitoring LAB distribution and persistence at the mucosal surfaces are needed. Here, we characterize Lactiplantibacillus plantarum bacteria expressing the infrared fluorescent protein IRFP713 for exploring their in vivo distribution in the mucosa and potential use as a mucosal vaccine vector. This bacterial species is commonly used as a vaginal probiotic and was recently found to have a niche in the human nose. Three different fluorescent L. plantarum strains were obtained using the nisin-inducible pNZRK-IRFP713 plasmid which contains the nisRK genes, showing stable and constitutive expression of IRFP713 in vitro. One of these strains was further monitored in BALB/c mice using near-infrared fluorescence, indicating successful colonization of the nasal and vaginal mucosae for up to 72 h. This study thus provides a tool for the in vivo spatiotemporal monitoring of lactiplantibacilli, allowing non-invasive bacterial detection in these mucosal sites. KEY POINTS: • Stable and constitutive expression of the IRFP713 protein was obtained in different L. plantarum strains. • IRFP713+ L. plantarum 3.12.1 was monitored in vivo using near-infrared fluorescence. • Residence times observed after intranasal and vaginal inoculation were 24-72 h.

Trace element profile and incidence of type 2 diabetes, cardiovascular disease and colorectal cancer: results from the EPIC-Potsdam cohort study.

PURPOSE: We aimed to examine the prospective association between manganese, iron, copper, zinc, iodine, selenium, selenoprotein P, free zinc, and their interplay, with incident type 2 diabetes (T2D), cardiovascular disease (CVD) and colorectal cancer (CRC). METHODS: Serum trace element (TE) concentrations were measured in a case-cohort study embedded within the EPIC-Potsdam cohort, consisting of a random sub-cohort (n = 2500) and incident cases of T2D (n = 705), CVD (n = 414), and CRC (n = 219). TE patterns were investigated using principal component analysis. Cox proportional hazard models were fitted to examine the association between TEs with T2D, CVD and CRC incidence. RESULTS: Higher manganese, zinc, iodine and selenium were associated with an increased risk of developing T2D (HR Q5 vs Q1: 1.56, 1.09-2.22; HR per SD, 95% CI 1.18, 1.05-1.33; 1.09, 1.01-1.17; 1.19, 1.06-1.34, respectively). Regarding CVD, manganese, copper and copper-to-zinc ratio were associated with an increased risk (HR per SD, 95% CI 1.13, 1.00-1.29; 1.22, 1.02-1.44; 1.18, 1.02-1.37, respectively). The opposite was observed for higher selenium-to-copper ratio (HR Q5 vs Q1, 95% CI 0.60, 0.39-0.93). Higher copper and zinc were associated with increasing risk of developing CRC (HR per SD, 95% CI 1.29, 1.05-1.59 and 1.14, 1.00-1.30, respectively). Selenium, selenoprotein P and selenium-to-copper-ratio were associated to decreased risk (HR per SD, 95% CI 0.82, 0.69-0.98; 0.81, 0.72-0.93; 0.77, 0.65-0.92, respectively). Two TE patterns were identified: manganese-iron-zinc and copper-iodine-selenium. CONCLUSION: Different TEs were associated with the risk of developing T2D, CVD and CRC. The contrasting associations found for selenium with T2D and CRC point towards differential disease-related pathways.

Parental education associated with immune function in adolescence.

BACKGROUND: The immune system is affected and shaped by several internal and external factors. Among the external variables, the socioeconomic status is known to influence the immune system since the early years of life and throughout life. METHODS: In this study, we assessed the relationship between parental education with the white blood cells and its subtypes in 1213 adolescents from the EPITeen cohort, assessed at the age of 13. Beta coefficients (ß) and 95% confidence intervals (CI) were fitted using linear regression models to quantify the association and were adjusted for sex, body mass index and chronic disease. RESULTS: After adjustment, parental education presented a negative association with white blood cells, which was significant among those with higher high-sensitivity C-reactive protein (hs-CRP) median levels [-0.05 mg/l (95% CI -0.08, -0.01)]. On the contrary, a positive association with lymphocytes was observed, which was, significant among those with lower hs-CRP [0.17 mg/l (95% CI 0.02, 0.32)]. A neutrophil-to-lymphocyte ratio significant decrease was also observed with the increment of parental education (P<0.001). CONCLUSION: We found that parental education was positively associated with a higher proportion of lymphocytes and a lower proportion of neutrophils, suggesting that parental education is associated with offsprinǵs innate immune system regulation. These results may contribute to clarify the relationships between childhood socioeconomic status and increased risk of adverse cardiovascular outcomes and other immune-related diseases.

Risk Factors and Prevention Strategies for Anxiety Disorders in Childhood and Adolescence.

Anxiety is prevalent in childhood and adolescence. Youth with maladaptive responses to common situations and stressors are at risk of having anxiety disorders. Persistent anxiety symptoms and anxiety disorders can be debilitating with long-term adverse outcomes in adulthood. Hence, decreasing the burden of anxiety disorders is an important public health priority. Development of anxiety disorders has a multifactorial etiology. There is a considerable complex interaction of genetics, temperament, parenting behavior, environmental triggers, and physiologic factors. Identification of these risk factors is key to early detection, prevention, and development of applicable management approaches. Despite several evidence-based treatments published, there are limited prevention strategies available. Effective implementation of prevention strategies is essential and can be achieved by either elimination or reduction of the negative risk factors or strengthening the protective factors on anxiety symptoms and anxiety disorders. This chapter reviews the common risk and protective factors and provides current literature on prevention strategies for pediatric and adolescent anxiety disorders.

Central and peripheral body fat distribution: Different associations with low-grade inflammation in young adults?

BACKGROUND AND AIMS: Evidence regarding the impact of regional body fat distribution on low-grade inflammation is limited. The current study examined the association of central and peripheral body fat distribution and low-grade inflammation levels in young adults, considering collinearity between variables. METHODS AND RESULTS: A cross-sectional analysis of 809 adults (aged 27 years) was conducted as part of the EPITeen cohort, Porto, Portugal. Regional body fat was measured by dual-energy X-ray absorptiometry scan (DXA) and serum high-sensitivity C-reactive protein (hsCRP) was measured in a fasting blood sample. OLS (ordinary least squares) and LASSO (least absolute shrinkage and selection operator) regression models were fitted to estimate the association of trunk and peripheral fat with hsCRP, stratified by sex. Using OLS regression, trunk fat in females was positively associated with ln(hsCRP) (ß1 = 0.064, 95% CI 0.018; 0.109). The effect of peripheral fat on ln(hsCRP) was shown not to be significantly different from trunk fat (ß2 = -0.011, 95% CI -0.110; 0.089), but no statistically significant association was observed (ß3 = 0.053, 95% CI -0.004; 0.110) between peripheral fat and ln(hsCRP). In males, trunk fat also showed a positive association with ln(hsCRP) (ß1 = 0.104, 95% CI 0.055; 0.154), and the effect of peripheral fat on ln(hsCRP) was shown to be significantly different from trunk fat (ß2 = -0.124, 95% CI -0.237;-0.011). However, the association between peripheral fat and ln(hsCRP) did not reach statistical significance (ß3 = -0.020, 95% CI -0.086; 0.046). The results of OLS were confirmed by LASSO regression. CONCLUSION: A higher fat deposited in the trunk was positively associated with hsCRP, whereas no statistically significant effect was observed for peripheral fat.

FTIR spectroscopy of chronic venous leg ulcer exudates: an approach to spectral healing marker identification.

Chronic venous leg ulcer (CVLU) arises as a chronic venous insufficiency complication and is a major cause of morbidity throughout the world. Our hypothesis is that the CVLU exudate composition is a biochemical representation of the wound clinical state. Then, Fourier Transform Infrared (FTIR) spectroscopy could be a useful and less-invasive technique to study the clinical state of the ulcer. For this, the aim of this work was to perform a spectral characterization of the exudate from CVLU using FTIR spectroscopy to identify potential healing markers. 45 exudate samples from CVLU, 95% of the strains isolated from CVLU in planktonic and biofilm phenotypes and other related biological samples such as human plasma, serum, urine, blood cells, urea, creatinine, glucose and albumin were studied by FTIR spectroscopy. According to the vibration frequency of biomolecules' (lipids, proteins, nucleic acids and carbohydrates) characteristic bonds in the infrared region, different spectral windows were selected and spectral areas of each window were measured. Besides, Savitzky-Golay second derivatives were obtained for all spectra and peaks from each standardized window were detected. FTIR spectroscopy allowed identification of sample types (exudate, plasma, serum, urine) as each one presents a unique relative composition and ratios range. Also, this technique could be useful to identify bacteria in the phenotypic-ulcer state and allows differentiation of whether bacteria are in the biofilm or planktonic form which is unlikely by conventional methods. In this work we found some spectral markers (areas, peaks) that allow identification of several parameters in the exudate such as (a) total cellularity, (b) inflammatory cell load, (c) bacterial load, (d) fibrin amount, and (e) inflammatory proteins. Because the measured areas or founded peaks are concentration-dependent this method could also serve to measure them. Therefore, FTIR spectroscopy could be useful to evaluate patient evolution as all these exudate parameters represent critical negative markers for wound healing.

Maternal Smoking: A Life Course Blood Pressure Determinant?

Introduction: Exposure to maternal smoking early in life may affect blood pressure (BP) control mechanisms. We examined the association between maternal smoking (before conception, during pregnancy, and 4 years after delivery) and BP in preschool children. Methods: We evaluated 4295 of Generation XXI children, recruited at birth in 2005-2006 and reevaluated at the age of 4. At birth, information was collected by face-to-face interview and additionally abstracted from clinical records. At 4-year follow-up, interviews were performed and children's BP measured. Linear regression models were fitted to estimate the association between maternal smoking and children's BP. Results: Children of smoking mothers presented significantly higher BP levels. After adjustment for maternal education, gestational hypertensive disorders, and child's body mass index, children exposed during pregnancy to maternal smoking presented a higher systolic BP (SBP) z-score (ß = 0.08, 95% confidence interval [CI] 0.04 to 0.14). In crude models, maternal smoking was associated with higher SBP z-score at every assessed period. However, after adjustment, an attenuation of the association estimates occurred (ß = 0.08, 95% CI 0.03 to 0.13 before conception; ß = 0.07, 95%CI 0.02 to 0.12; ß = 0.04, 95%CI -0.02 to 0.10; and ß = 0.06, 95%CI 0.00 to 0.13 for the first, second, and third pregnancy trimesters, respectively; and ß = 0.07, 95%CI 0.02 to 0.12 for current maternal smoking). No significant association was observed for diastolic BP z-score levels. Conclusion: Maternal smoking before, during, and after pregnancy was independently associated with systolic BP z-score in preschool children. This study provides additional evidence to the public health relevance of maternal smoking cessation programs if early cardiovascular health of children is envisaged. Implications: Using observational longitudinal data from the birth cohort Generation XXI, this study showed that exposure to maternal smoking-before pregnancy, during pregnancy, and 4 years after delivery-was associated with a systolic BP-raising effect in children at the age of 4. The findings of this study add an important insight into the need to support maternal smoke-free environments in order to provide long-term cardiovascular benefit, starting as early as possible in life.

Relationship between dietary vitamin D and serum 25-hydroxyvitamin D levels in Portuguese adolescents.

OBJECTIVE: To understand the relationship between vitamin D intake and serum 25-hydroxyvitamin D (25(OH)D) levels in a sample of Portuguese adolescents. DESIGN: Cross-sectional evaluation carried out in the 2003/2004 school year. Vitamin D intake was assessed by an FFQ and 25(OH)D was measured in a fasting blood sample. SETTING: Public and private schools in Porto, Portugal. SUBJECTS: Adolescents aged 13 years (n 521) enrolled at school (EPITeen cohort). RESULTS: Both mean (sd) intake and serum 25(OH)D level were far below the recommended, 4·47 (2·49) µg/d and 16·5 (5·7) ng/ml, respectively. A significant difference in serum level was found according to season, with lower values in winter than summer (14·8 (4·6) v. 17·3 (5·9) ng/ml, P<0·001). Vitamin D intake was weakly correlated with serum 25(OH)D (r=0·056, P=0·203). CONCLUSIONS: Dietary vitamin D and serum 25(OH)D levels were positively but weakly correlated and the error was higher among those with higher serum 25(OH)D concentration. Our results support the need for strategies that promote increase of the most important food sources of vitamin D to reduce the high prevalence of low vitamin D status.

Sociodemographic characteristics determine dietary pattern adherence during pregnancy.

OBJECTIVE: Sociodemographic factors may affect adherence to specific dietary patterns during pregnancy. The present study aimed to identify dietary patterns during pregnancy and associated factors among Brazilian pregnant women. DESIGN: A cross-sectional analysis. Dietary intake was evaluated with a semi-quantitative FFQ during the first postpartum week; the time frame included the second and third gestational trimesters. Principal component analysis was used to identify dietary patterns during pregnancy. Sociodemographic data were obtained using a structured questionnaire. Multiple linear regressions were applied to test the associations between the sociodemographic factors and dietary patterns. SETTING: Mesquita, Rio de Janeiro, Brazil, 2011. SUBJECTS: Postpartum women (n 327) who were 18-45 years of age and Mesquita residents. RESULTS: Three different dietary patterns were identified: 'healthy' (mainly comprising legumes, vegetables and fruits), 'mixed' (mainly comprising candy, butter and margarine, and snacks) and 'traditional' (mainly comprising beans and rice). Women with a higher monthly per capita family income (ß=0·0006; 95% CI 0·0001, 0·001; P=0·011) and women of older age (ß=0·021; 95% CI -0·001, 0·042; P=0·058) were more likely to adhere to the 'healthy' dietary pattern. Women with higher parity were less likely to adhere to the 'healthy' pattern (ß=-0·097; 95% CI -0·184, -0·009; P=0·030) and were more likely to adhere to the 'traditional' pattern (ß=0·098; 95% CI 0·021, 0·175; P=0·012). Although not statistically significant, older women were less likely to adhere to the 'mixed' (ß=-0·017; 95% CI -0·037, 0·003; P=0·075) and 'traditional' (ß=-0·018; 95% CI -0·037, 0·001; P=0·061) dietary patterns. CONCLUSIONS: Monthly per capita family income, parity and maternal age were factors associated with adherence to a healthy diet during pregnancy.

Novel topical formulation for ischemic chronic wounds. Technological design, quality control and safety evaluation.

Ulceration of the foot in diabetes is common and disabling, and frequently leads to amputation of the leg. The pathogenesis of foot ulceration is complex, clinical presentation variable and management requires early expert assessment. Despite treatment, ulcers readily become chronic wounds. Chronic wounds are those that remain in a chronic inflammatory state failing a normal healing process patterns. This is partially caused by inefficient eradication of opportunistic pathogens like Pseudomonas aeruginosa. We propose its control or eradication will promote wound healing. Lactobacillus plantarum cultures supernatants (LAPS) shows antipathogenic and pro-healing properties. The main objective was to design two pharmaceutical dosage forms by using LAPS as active pharmaceutical ingredient and to perform its quality control, in vitro activity conservation tests and human trials (safety evaluation). Both selected formulations reach the technological quality expected for 120 days, shows adequate occlusive characteristics and proper adhesion to human skin. From the in vitro release assays were found that LAPS shows adequate release from matrix and maintain its antimicrobial and anti-biofilm activity. First human trials were developed and neither edema nor erythema on healthy skin voluntaries was found. We conclude that C80 and C100 are adequate for their use in future clinical trials to demonstrate a comprehensive therapeutic effectiveness in ischemic chronic wounds.

Equipment and method for in vitro release measurements on topical dosage forms.

CONTEXT: In countries where research budgets are meager as Argentina, the tendency to innovation and improvements in the designs prototypes "made in Argentina" marks a growing trend adopted by researchers. This article presents a diffusion cell of original design, for release studies of Active Pharmaceutical Ingredient (API) from classical topical dosage forms, also includes the methodology for its optimization and validation. The objective was to evaluate and validate a system designed and to compare it to the Franz cells system. METHODS: Parameters, reproducibility and robustness were performed included factors as, stirring conditions, membrane stabilization treatment and temperature variation. Release and retention on membrane assay were performed using two different API and formulations. RESULTS: The method is reproducible and robust for the parameters tested. Release assays show that no significative difference with the Franz Cells system. Our system allows the simultaneous measurement of different parameters, representing an innovation on these methodologies. The LMC was used for assays of in vitro retention on membrane and the values obtained were reproducible and coincident whit values obtained for other authors. CONCLUSIONS: The system designed and the methodology employed, are acceptable for in vitro release studies. The device and method has the characteristics required.

Formulation and quality control of semi-solid containing harmless bacteria by-products: chronic wounds pro-healing activity.

Chronic wounds are those that remain in a chronic inflammatory state and fail to follow normal healing process. Infection is one of the most important causes of chronicity. A frequent pathogen isolated from chronic infections is Pseudomonas aeruginosa; refractory to therapy and host immune attack in its biofilm phenotype. Lactobacillus plantarum cultures supernatants (LAPS) interfere with its pathogenic capacity. In addition, LAPS showed bacteriostatic and bactericide properties and is neither cytotoxic nor an inductor of necrosis-apoptosis. LAPSs chemical composition was determined; allowing us to propose a correlation between its constituents and their biological activity. This article shows a pharmaceutical dosage form designed by using LAPS as an API with pro-healing activity and its quality control. Pharmacotechnical and anti-microbial assays were adapted to demonstrate that the vehicle used does not modify LAPS activities. Selected formulation (F100) showed fair mechanical and technological properties. From the in vitro release assays was found an adequate release from the carrier matrix and maintains its anti-pathogenic activity for 6 months. We propose F100 for chronic wounds treatment. The use of harmless bacteria by-products, such as LAPS, to antagonize infectious pathogens that have ability to form biofilm is an efficient and economic approach to treat infected chronic wounds.

Compounds from Lactobacillus plantarum culture supernatants with potential pro-healing and anti-pathogenic properties in skin chronic wounds.

CONTEXT: It is necessary to advance the field of alternative treatments for chronic wounds that are financially accessible to the least economically developed countries. Previously we demonstrated that topical applications of Lactobacillus plantarum culture supernatants (LAPS) on human-infected chronic wounds reduce the pathogenic bioburden, the amount of necrotic tissue, and the wound area, as well as promote debridement, granulation tissue, and wound healing. OBJECTIVE: To study LAPS chemically and biologically and to find potential molecules responsible for its pro-healing and anti-pathogenic properties in chronic wounds. MATERIALS AND METHODS: (1) Chemical analysis: extracts were subjected to a column chromatography and the fractions obtained were studied by GCMS. (2) Quantification: dl-lactic acid (commercial kit), phenolic compounds (Folin-Ciocalteu), H2O2 (micro-titration), and cations (flame photometry). (3) Biological analysis: autoinducers type 2 (AI-2) (Vibrio harveyi BB170 bioassay), DNAase activity (Agar DNAase), and Pseudomonas aeruginosa biofilm inhibition (crystal violet technique). RESULTS: According to its biological activity, the most significant molecules found by GCMS were the following: antimicrobials (mevalonolactone, 5-methyl-hydantoine, benzoic acid, etc.); surfactants (di-palmitin, distearin, and 1,5-monolinolein); anesthetics (barbituric acid derivatives), and AI-2 precursors (4,5-dihydroxy-2,3-pentanedione and 2-methyl-2,3,3,4-tetrahydroxytetrahydrofurane). Concentrations measured (µg/mL): DL-lactic acid (11.71 ± 1.53) and H2O2 (36 ± 2.0); phenolic compounds (485.2 ± 15.20); sodium (370 ± 17); potassium 920 ± 24); calcium (20 ± 4); and magnesium (15 ± 3). DNAase from LAPS had activity on genomic DNA from PMNs and P. aeruginosa. DISCUSSION AND CONCLUSION: The molecules and biological activities found in LAPS could explain the observed effects in human chronic wounds.

Efficacy of coupling repetitive transcranial magnetic stimulation and physical therapy to reduce upper-limb spasticity in patients with stroke: a randomized controlled trial.

OBJECTIVE: To assess the efficacy of inhibitory repetitive transcranial magnetic stimulation (rTMS) for decreasing upper-limb muscle tone after chronic stroke. DESIGN: A randomized sham-controlled trial with a 4-week follow-up. SETTING: Research hospital. PARTICIPANTS: Patients with stroke (N=20) with poststroke upper limb spasticity. INTERVENTIONS: The experimental group received rTMS to the primary motor cortex of the unaffected side (1500 pulses; 1Hz; 90% of resting motor threshold for the first dorsal interosseous muscle) in 10 sessions, 3d/wk, and physical therapy (PT). The control group received sham stimulation and PT. MAIN OUTCOME MEASURES: Modified Ashworth scale (MAS), upper-extremity Fugl-Meyer assessment, FIM, range of motion, and stroke-specific quality-of-life scale. All outcomes were measured at baseline, after treatment (postintervention), and at a 4-week follow-up. A clinically important difference was defined as a reduction of ≥1 in the MAS score. RESULTS: Friedman test revealed that PT is efficient for significantly reducing the upper limb spasticity of patients only when it is associated with rTMS. In the experimental group, 90% of the patients at postintervention and 55.5% at follow-up showed a decrease of ≥1 in the MAS score, representing clinically important differences. In the control group, 30% of the patients at postintervention and 22.2% at follow-up experienced clinically meaningful changes. There were no differences between the groups at any time for any of the other outcome measures, indicating that both groups demonstrated similar behaviors over time for all variables. CONCLUSIONS: rTMS associated with PT can be beneficial in reducing poststroke spasticity. However, more studies are needed to clarify the clinical changes underlying the reduction in spasticity induced by noninvasive brain stimulations.

[15]aneN4S: synthesis, thermodynamic studies and potential applications in chelation therapy.

The purpose of this work was to synthesize and characterize the thiatetraaza macrocycle 1-thia-4,7,10,13-tetraazacyclopentadecane ([15]aneN4S). Its acid-base behaviour was studied by potentiometry at 25 °C and ionic strength 0.10 M in KNO3. The protonation sequence of this ligand was investigated by 1H-NMR titration that also allowed the determination of protonation constants in D2O. Binding studies of [15]aneN4S with Mn2+, Fe2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+, Hg2+ and Pb2+ metal ions were further performed under the same experimental conditions. The results demonstrated that this compound has a higher selectivity and thermodynamic stability for Hg2+ and Cu2+, followed by Ni2+. The UV-visible-near IR spectroscopies and magnetic moment data for the Co(II) and Ni(II) complexes indicated a tetragonal distorted coordination geometry for both metal centres. The value of magnetic moment and the X-band EPR spectra of the Cu(II) complex are consistent with a distorted square pyramidal geometry.

Neph1 is required for neurite branching and is negatively regulated by the PRRXL1 homeodomain factor in the developing spinal cord dorsal horn.

The cell-adhesion molecule NEPH1 is required for maintaining the structural integrity and function of the glomerulus in the kidneys. In the nervous system of Drosophila and C. elegans, it is involved in synaptogenesis and axon branching, which are essential for establishing functional circuits. In the mammalian nervous system, the expression regulation and function of Neph1 has barely been explored. In this study, we provide a spatiotemporal characterization of Neph1 expression in mouse dorsal root ganglia (DRGs) and spinal cord. After the neurogenic phase, Neph1 is broadly expressed in the DRGs and in their putative targets at the dorsal horn of the spinal cord, comprising both GABAergic and glutamatergic neurons. Interestingly, we found that PRRXL1, a homeodomain transcription factor that is required for proper establishment of the DRG-spinal cord circuit, prevents a premature expression of Neph1 in the superficial laminae of the dorsal spinal cord at E14.5, but has no regulatory effect on the DRGs or on either structure at E16.5. By chromatin immunoprecipitation analysis of the dorsal spinal cord, we identified four PRRXL1-bound regions within the Neph1 introns, suggesting that PRRXL1 directly regulates Neph1 transcription. We also showed that Neph1 is required for branching, especially at distal neurites. Together, our work showed that Prrxl1 prevents the early expression of Neph1 in the superficial dorsal horn, suggesting that Neph1 might function as a downstream effector gene for proper assembly of the DRG-spinal nociceptive circuit.