Biodistribution and intracellular localization of hyaluronan and its nanogels. A strategy to target intracellular S. aureus in persistent skin infections.

Intracellular pathogens are a critical challenge for antimicrobial therapies. Staphylococcus aureus (S. aureus) causes approximately 85% of all skin and soft tissue infections in humans worldwide and more than 30% of patients develop chronic or recurrent infections within three months, even after appropriate antibacterial therapies. S. aureus is also one of the most common bacteria found in chronic wounds. Recent evidences suggest that S. aureus is able to persist within phagolysosomes of skin cells (i.e. keratinocytes, phagocytic cells), being protected from both the immune system and a number of antimicrobials. To overcome these limits, nano-formulations that enable targeted therapies against intracellular S. aureus might be developed. Herein, the biodistribution and intracellular localisation of hyaluronan (HA) and HA-based nanoparticles (nanogels, NHs) are investigated, both after intravenous (i.v.) injections (in mice) and topical administrations (in ex vivo human skin). Results indicate HA and NHs accumulate especially in skin and liver of mice after i.v. injection. After topical application on human skin explants, no penetration of both HA and NHs was detected in skin with intact stratum corneum. By contrast, in barrier-disrupted human skin (with partial removal and loosening of stratum corneum), HA and NHs penetrate to the viable epidermis and are taken up by keratinocytes. In mechanically produced wounds (skin without epidermis) they accumulate in wound tissue and are taken up by dermis cells, e.g. fibroblasts and phagocytic cells. Interestingly, in all cases, the cellular uptake is CD44-mediated. In vitro studies confirmed that after CD44-mediated uptake, both HA and NHs accumulate in lysosomes of dermal fibroblasts and macrophages, as previously reported for keratinocytes. Finally, the colocalisation between intracellular S. aureus and HA or NHs is demonstrated, in macrophages. Altogether, for the first time, these results strongly suggest that HA and HA-based NHs can provide a targeted therapy to intracellular S. aureus, in persistent skin or wound infections.

Halting hyaluronidase activity with hyaluronan-based nanohydrogels: development of versatile injectable formulations.

Hyaluronan (HA) is among the most used biopolymers for viscosupplementation and dermocosmetics. However, the current injectable HA-based formulations present relevant limitations: I) unmodified HA is quickly degraded by endogenous hyaluronidases (HAase), resulting in short lasting properties; II) cross-linked HA, although shows enhanced stability against HAase, often contains toxic chemical cross-linkers. As such, herein, we present biocompatible self-assembled hyaluronan-cholesterol nanohydrogels (HA-CH NHs) able to bind to HAase and inhibit the enzyme activity in vitro, more efficiently than currently marketed HA-based cross-linked formulations (e.g. Jonexa™). HA-CH NHs inhibit HAase through a mixed mechanism, by which NHs bind to HAase with an affinity constant 7-fold higher than that of native HA. Similar NHs, based on gellan-CH, evidenced no binding to HAase, neither inhibition of the enzyme activity, suggesting this effect might be due to the specific binding of HA-CH to the active site of the enzyme. Therefore, HA-CH NHs were engineered into injectable hybrid HA mixtures or physical hydrogels, able to halt the enzymatic degradation of HA.

Erythrocyte's aging in microgravity highlights how environmental stimuli shape metabolism and morphology.

The determination of the function of cells in zero-gravity conditions is a subject of interest in many different research fields. Due to their metabolic unicity, the characterization of the behaviour of erythrocytes maintained in prolonged microgravity conditions is of particular importance. Here, we used a 3D-clinostat to assess the microgravity-induced modifications of the structure and function of these cells, by investigating how they translate these peculiar mechanical stimuli into modifications, with potential clinical interest, of the biochemical pathways and the aging processes. We compared the erythrocyte's structural parameters and selected metabolic indicators that are characteristic of the aging in microgravity and standard static incubation conditions. The results suggest that, at first, human erythrocytes react to external stimuli by adapting their metabolic patterns and the rate of consumption of the cell resources. On longer timeframes, the cells translate even small differences in the environment mechanical solicitations into structural and morphologic features, leading to distinctive morphological patterns of aging.

HLA-DRB1*15 association with multiple sclerosis is confirmed in a multigenerational Italian family.

Environmental and genetic factors seem to play a pathogenetic role in multiple sclerosis (MS). The genetic component is partly suggested by familial aggregation of cases; however, MS families with affected subjects over different generations have rarely been described. The aim of this study was to report clinical and genetic features of a multigenerational MS family and to perform a review of the literature on this topic. We describe a multigenerational Italian family with six individuals affected by MS, showing different clinical and neuroradiological findings. HLA-DRB1* typing revealed the presence of the DRB1*15:01 allele in all the MS cases and in 4/5 non-affected subjects. Reports on six multigenerational MS families have previously been published, giving similar results. The HLA-DRB1*15:01 allele was confirmed to be linked to MS disease in this family; moreover, its presence in non-affected subjects suggests the involvement of other susceptibility factors in the development and expression of the disease, in accordance with the complex disease model now attributed to MS.

The impact of excess body fat on bone remodeling in adolescents.

The impact of excess body fat on bone remodeling was evaluated in overweight, obese, and extremely obese adolescents. In adolescents with excess weight, it was observed that the higher the bone mineral content and bone mineral density values, the lower the levels of the biomarkers. Nutritional imbalances by excess had a negative effect on bone formation in this stage of life. INTRODUCTION: The aim of this study was to investigate the impact of excess body fat on bone remodeling in adolescents. METHODS: Body weight, height, and body mass index were determined in 391 adolescents classified as normal weight, overweight, obese, and extremely obese. Bone age was obtained and bone mineral content and bone mineral density were evaluated in the lumbar spine, proximal femur, and total and subtotal body. Blood samples were collected for evaluation of the following bone biomarkers: osteocalcin, bone alkaline phosphatase (BAP), and serum carboxy-terminal telopeptide (S-CTx). The data were analyzed according to nutritional status and age. RESULTS: In girls with excess weight, the biomarkers were higher in the 10 to 13-year age group and no significant differences were observed between groups according to nutritional status. In boys, the levels were higher in those aged 13 to 15 years. According to nutritional status, significant differences were only observed in mean S-CTx for the age groups of 10-15 years, with higher levels between overweight and obese adolescents aged 10-12 years and between obese and extremely obese adolescents aged 13-15 years. In girls, significant negative correlations were observed between lean mass, fat mass, and fat percentage and each of the three bone markers studied. There was no correlation between lean mass or fat mass and the three biomarkers in boys. The biomarker trends demonstrated across the age groups follow the age trends for growth velocity. CONCLUSIONS: The higher the fat percentage and fat mass in girls, the lower the levels of the biomarkers, indicating that excess body fat has a negative effect on the evolution of these markers during adolescence.

Endometrial cancer in unicornuate uterus: a case report.

PURPOSE OF INVESTIGATION: Miillerian anomalies have not been implicated as a significant risk factor for the development of cervical, uterine, and ovarian cancers; in the present literature, there are only a few reports of endometrial cancer arising in patients with Miillerian abnormalities. To the best of the authors' knowledge, this is the first reported case of endometrial cancer arising in a patient with unicornuate uterus. CASE REPORT: A 69-year-old Caucasian woman underwent clinical examination and office hysteroscopy with endometrial biopsy because of abnormal post-menopausal bleeding. The diagnosis was endometrial cancer in unicornuate uterus, hence the patient underwent total hysterectomy with pelvic lymphadenectomy. CONCLUSION: Uterine malformations and genetic disorders may cause a delayed diagnosis of gynaecological cancers. Gynaecological examination in asymptomatic patients and differential diagnosis in abnormal uterine bleeding patients should be considered.

Novel FUS mutations identified through molecular screening in a large cohort of familial and sporadic amyotrophic lateral sclerosis.

BACKGROUND AND PURPOSE: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. Approximately 5%-10% of cases are familial (FALS) and the remaining are sporadic (SALS). To date FUS mutations are responsible for 4%-6% of familial cases as well as 0.7%-1.8% of sporadic cases. METHODS: The frequency of FUS mutations was investigated in an Italian cohort of 500 SALS and 40 FALS patients through direct sequencing of exons 5, 6, 13, 14 and 15. RESULTS: Eight FUS mutation carriers were identified in five SALS (1%) and three FALS (7.5%), five already known and three new mutations: a de novo mutation was identified in a sporadic subject as well as the co-presence of FUS/C9ORF72 mutations in a FALS subject. The molecular and clinical details of the three patients harbouring a novel mutation (G245V, G509D and R491C) are presented here. Moreover the co-presence of the R491C mutation and C9ORF72 pathological expansion was found according to the oligogenic disease model. CONCLUSIONS: In conclusion our results revealed a higher frequency of FUS mutation carriers (7.5%) in FALS compared to literature data together with a higher presence of female gender.

Benign pulmonary metastasizing leiomyomatosis: case report.

The authors report a rare case of leiomyomatosis of the lung diagnosed in a 43-year-old woman, with uterine intravenous leiomyomatosis. Benign metastasizing leiomyoma (BML) is an extremely rare lesion characterized by usually multiple, benign-appearing smooth muscle tumors of the lung in females with coexisting uterine leiomyoma. On the basis of their histological and immunohistological features, a unified histogenetic view of leiomyomas with vascular invasion (LWVI) and BML of the uterus is proposed. LWVI and BML may be the same pathological entity and microscopic vascular invasion may represent the metastatic mechanism of BML. LWVI seems to be the precursor of BML.

Amyotrophic lateral sclerosis in pregnancy is associated with a vascular endothelial growth factor promoter genotype.

BACKGROUND AND PURPOSE: The occurrence of amyotrophic lateral sclerosis (ALS) during pregnancy is uncommon and the effect of one on the other is not well described. METHODS: The clinical and genetic features of five cases of ALS are reported with an onset during pregnancy or within 1 month from delivery. Charts from 239 women with a diagnosis of ALS attending the neuromuscular clinics at the Neuromuscular Omnicentre (NEMO) and at IRCCS Policlinico San Donato from 2008 to 2011 were reviewed. RESULTS: Of these, 12.8% of the women in child-bearing age had a diagnosis of ALS during pregnancy or immediately after delivery. Genetic screening of the major causative genes revealed two mutations in superoxide dismutase 1 (SOD1) gene; the analysis of vascular endothelial growth factor (VEGF) promoter variation showed a segregation of the haplotype CA/AG (-2578C/A; -1190A/G) in patients developing ALS related to pregnancy. No effects on foetal development or neonatal course were observed. CONCLUSIONS: Pregnancy may unmask ALS but whether this is coincidental is unclear. Hormonal and inflammatory modifications might trigger ALS in subjects with increased susceptibility to oxidative stress related to the toxic function of SOD1 or in subjects with a reduction of neuroprotective molecules such as VEGF.

Reduction in dietary trans fat intake is associated with decreased LDL particle number in a primary prevention population.

BACKGROUND AND AIMS: Increased trans fat intake has been associated with an increased risk of cardiovascular disease (CVD). While the effect of trans fat on traditional lipids is known, it's association with LDL particle number (LDL-P), a novel marker of CVD risk, has not been established. The purpose of this study was to determine the association between trans fat intake and LDL-P over 1-year among individuals participating in a lifestyle intervention trial. METHODS AND RESULTS: Family members (N = 400, 33% male, mean age 48 ± 13) of patients hospitalized with CVD who participated in a 1-year randomized controlled primary prevention lifestyle intervention trial and had complete dietary data and LDL-P measures at baseline and 1-year. Change in trans fat as a percentage of total diet and mean absolute change in LDL-P at 1-year was assessed using multivariate adjusted linear regression models. At baseline, there was a significant positive correlation between dietary trans fat intake and LDL-P (Beta = 37, p = 0.04). For every 1 percent change in trans fat intake there was a 27 nmol/L change in LDL-P (Beta = 27, p = 0.04) over 1-year which was independent of baseline predictors and confounders (age, sex, smoking, statin use, waist size and physical activity; Beta = 30, p = 0.03). CONCLUSION: A reduction in trans fat intake over 1-year was significantly associated with a reduction in LDL-P independent of potential confounders. Healthcare providers should reinforce the beneficial impact of a healthy diet, and in particular modifications in trans fat intake on improving lipid profiles.

Vaginal microbiota and viral sexually transmitted diseases.

Healthy vaginal microbiota is an important biological barrier to pathogenic microorganisms. When this predominantly Lactobacillus community is disrupted, decreased in abundance and replaced by different anaerobes, bacterial vaginosis (BV) may occur. BV is associated with prevalence and incidence of several sexually transmitted infections. This review provides background on BV, discusses the epidemiologic data to support a role of altered vaginal microbiota for acquisition of sexually transmitted diseases and analyzes mechanisms by which lactobacilli could counteract sexually transmitted viral infections.

The expression pattern of small nucleolar and small Cajal body-specific RNAs characterizes distinct molecular subtypes of multiple myeloma.

Small nucleolar RNAs (snoRNAs) and small Cajal body-specific RNAs (scaRNAs) are non-coding RNAs involved in the maturation of other RNA molecules and generally located in the introns of host genes. It is now emerging that altered sno/scaRNAs expression may have a pathological role in cancer. This study elucidates the patterns of sno/scaRNAs expression in multiple myeloma (MM) by profiling purified malignant plasma cells from 55 MMs, 8 secondary plasma cell leukemias (sPCLs) and 4 normal controls. Overall, a global sno/scaRNAs downregulation was found in MMs and, even more, in sPCLs compared with normal plasma cells. Whereas SCARNA22 resulted the only sno/scaRNA characterizing the translocation/cyclin D4 (TC4) MM, TC2 group displayed a distinct sno/scaRNA signature overexpressing members of SNORD115 and SNORD116 families located in a region finely regulated by an imprinting center at 15q11, which, however, resulted overall hypomethylated in MMs independently of the SNORD115 and SNORD116 expression levels. Finally, integrative analyses with available gene expression and genome-wide data revealed the occurrence of significant sno/scaRNAs/host genes co-expression and the putative influence of allelic imbalances on specific snoRNAs expression. Our data extend the current view of sno/scaRNAs deregulation in cancer and add novel information to the bio-molecular complexity of plasma cell dyscrasias.

Effectiveness of Lactobacillus-containing vaginal tablets in the treatment of symptomatic bacterial vaginosis.

The purpose of this study was to determine the effectiveness of Lactobacillus-containing vaginal tablets in the treatment of bacterial vaginosis (BV) and in the restoration of a healthy vaginal flora. Thirty-nine women with BV were enrolled in a double-blind, placebo-controlled clinical trial. Patients received either one Lactobacillus-containing tablet or placebo daily for 7 days. Clinical criteria, vaginal Gram stain scores and symptoms were compared with those at the initial visit and those at completion of therapy and 2 weeks later. After completion of therapy, all of the patients in the Lactobacillus-treated group (n = 18) were free of BV, showing a normal (83%) or intermediate (17%) vaginal flora, as compared with only two patients free of BV with intermediate flora (12%) from among the 16 placebo-treated women (p <0.001). Two weeks after completion of therapy, treatment was successful (score <7) in 61% of Lactobacillus-treated patients as compared with 19% of those in the placebo group (p <0.05). In the treatment group, the total number of symptomatic patients and the intensity of their symptoms, in particular vaginal malodour, were significantly reduced at both follow-up visits. The data indicate that intravaginal administration of exogenous selected strains of lactobacilli can restore a normal vaginal microbiota and be used in treating bacterial vaginosis.

Unusual coexistence of bilateral keratoconus and optic disc pit: a case report.

PURPOSE: To report the unknown coexistence of bilateral optic disc pit and keratoconus. METHODS: A 23-year-old man with bilateral keratoconus underwent complete ophthalmology screening, with an unexpected detection of undiagnosed optic disc pit in both eyes. Computerized corneal topography (CT), Orbscan, corneal pachometry, endothelial microscopy, and optical coherence tomography (OCT) examination were performed. RESULTS: The corneal CT showed a keratoconus pattern in both eyes, evolved in the right eye with a minimum corneal pachometry of 336 micronm in the right eye and 405 micronm in the left eye. Mean endothelial cell density was 1937 cells/mm2 in the right eye and 1912 cells/mm2 in the left eye. The OCT scans showed the presence of the disc pit in both eyes with a normal macular thickness and profile in the right eye, and in the left eye an augmented retinal thickness in the nasal macular zone due to retinal oedema and schisis, with an initial detachment of the neuroepithelium in the parapapillary area starting from the optic pit. CONCLUSIONS: This is the first clinical report of bilateral optic disc pit and keratoconus. Further investigations will be necessary to assess if there is a possible pathogenetic correlation between these two ocular pathologies or if this is an unusual coexistence of separate entities.

Raloxifeno aumenta el riesgo de ictus en mujeres de alto riesgo / Effects of Raloxifene on cardiovascular events and breast

No disponible

Myocardial blood flow and flow reserve in response to hormone therapy in postmenopausal women with risk factors for coronary disease.

Estrogen has beneficial effects on markers of coronary heart disease (CHD) risk, but may increase overall CHD events. The effects of hormone therapy on vascular endothelial function have been mixed, and require further assessment. We studied the myocardial blood flow (MBF) response to postmenopausal combination hormone therapy (CHT) in postmenopausal women with risk factors for CHD. We performed dynamic [13N]ammonia positron emission tomography in 15 postmenopausal women in a 7-month placebo-controlled crossover trial of continuous conjugated equine estrogen/cyclical micronized progesterone. MBF was measured at rest, after sympathetic stimulation with the cold pressor test (CPT), and after i.v. adenosine infusion, to determine baseline, endothelium-dependent, and maximal flows, respectively. Response to CPT was neutral in all women at baseline (-0.51 +/- 27%). Adenosine induced a marked increase in MBF (161 +/- 111%). Treatment with 3 months of combined estrogen/progestin CHT did not change CPT or adenosine MBF responses. Myocardial flow reserve was unchanged as well. In this group of postmenopausal women at higher cardiovascular risk, no association was found between CHT assignment and change in MBF. Further study is needed to clarify the effects of CHT on the endothelium of women with presumably diseased vasculature.

Association between self-reported physical activity and vascular reactivity in postmenopausal women.

Short-term exercise training has been associated with improved endothelial-dependent vasodilation, but the impact of long-term habitual physical activity on vascular reactivity is not established. We studied the correlation between self-reported, habitual physical activity and vasoreactivity in non-smoking, non-diabetic, postmenopausal women (n=34, mean age 65.6+/-7.4 years). Vasoreactivity was evaluated by the percentage and absolute change in brachial artery diameter in response to reactive hyperemia induced by occlusion-release, and in response to cold pressor testing (CPT). Habitual physical activity was assessed by a standardized questionnaire based on participant recall. Our results indicate that 64.7% of the women were exercising-to-sweat > or =1x/week, 4.8 flights of stairs were climbed/day, 5.0 city blocks were walked/day and 29.4% participated in moderately physically demanding daily activity. There was a significant association between the number of city blocks walked daily and exercising-to-sweat > or =1x/week with brachial artery percentage and absolute change to CPT (P<0.05). Women who reported a moderately physically demanding daily activity had a significantly greater brachial reactivity percentage change in response to CPT compared with those performing less demanding daily activity (2.0+/-3.6 versus 1.4+/-7.0%, P<0.05). The response to reactive hyperemia was also greater in those women reporting moderately physically demanding daily activity compared to less active women (6.5+/-5.4 versus 5.8+/-5.9%, P=n.s.), but this did not reach statistical significance. Stepwise, multivariate analysis adjusting for body mass index and HDL-cholesterol eliminated the association between physical activity and brachial reactivity in response to CPT, suggesting that physical activity may affect vasoreactivity via these mechanisms. This study suggests that moderate levels of self-reported physical activity are associated with a greater brachial reactivity in response to CPT and supports the recommendation that moderate intensity physical activity may be cardioprotective in postmenopausal women.

An evaluation of Choose to Move 1999: an American Heart Association physical activity program for women.

BACKGROUND: Rates of physical inactivity and poor nutrition, which are 2 of the most important modifiable risk factors for cardiovascular disease in women, are substantial. Even so, studies of interventions designed to improve lifestyle behaviors in women have been limited and often confined to particular geographical areas. OBJECTIVE: To evaluate the effect of Choose to Move on increasing women's physical activity, improving their knowledge of heart disease and stroke, and improving their nutrition. PARTICIPANTS AND METHODS: A prospective, nonrandomized, 12-week educational intervention designed by the American Heart Association for women across the United States. Participants received a welcome kit and manual with weekly information about how to manage cardiovascular disease risk factors and how to build a support system for lifestyle change. Women (N = 23 171) aged 25 years or older were recruited by direct mail, the media, health care providers, and other means. Follow-up evaluations were returned from 6389 women at 2 weeks, 5338 at 4 weeks, 4209 at 8 weeks, 3916 at 10 weeks, and 3775 at 12 weeks. Participants self-reported their physical activity, diet, and knowledge about heart disease, stroke, and related symptoms. RESULTS: Ninety percent of the participants were white and 56% were aged between 35 and 54 years. Among the participants who completed the week 12 follow-up evaluation, the percentage who reported being active (at least moderate exercise > or =5 times per week or >2(1/2) hours per week for the past 1 to 6 months) increased from 32% at baseline to 67% at the program's end (P =.001). Participants currently limiting excess calories or fat increased from 72% to 91% at week 10 follow-up evaluation (P =.001). The proportion correctly identifying heart disease as the leading cause of death increased from 84% to 91% at week 10 follow-up evaluation (P<.001). CONCLUSIONS: Women who completed the Choose to Move program evaluation reported that they significantly increased their levels of physical activity, reduced their consumption of high-fat foods, and increased their knowledge and awareness of cardiovascular disease risk and its symptoms. This program provides an important model for public health, voluntary, and other health organizations of population-based, targeted low-cost self-help programs that support the Healthy People 2010 objectives for physical activity, nutrition, and cardiovascular health.

Variability in fibrinogen measurements: an obstacle to cardiovascular risk stratification.

The clinical utility of fibrinogen measurement has been limited by large intraindividual variability. Several approaches that have been shown to improve the repeatability of fibrinogen include acquisition of samples at the same time of day, standardized sample procurement techniques, and multiple replicate sampling. This study employed established pre-analytical and analytical techniques known to reduce fibrinogen variability, including the acquisition of three replicate samples, each analyzed in duplicate, to evaluate the impact of intraindividual variability in fibrinogen measurement at baseline and 3 months on cardiovascular risk in 60 healthy subjects. Classification accuracy was evaluated by the ability to categorize subjects into tertiles of fibrinogen. Only 55% (33/60) of the subjects were correctly assigned to the appropriate fibrinogen tertile. Fibrinogen measurements varied by more than 10% in 45% of subjects and by 5% in 80% of subjects. Intraindividual variability in fibrinogen measurement with a functional assay limits cardiovascular risk assessment even when three replicates are averaged.