Antimicrobial activity and biofilm inhibition of riparins I, II and III and ultrastructural changes in multidrug-resistant bacteria of medical importance.

Natural products have been used to treat various infections; however, the development of antimicrobials has made natural products in disuse. Riparin I, II and III are natural alkamide isolated from Aniba riparia (Ness) Mez (Lauraceae), that exhibit economic importance and it is used in traditional medicine, and popularly known as "louro". This study investigated the cytotoxicity, antimicrobial and antibiofilm activity, and ultrastructural changes in vitro by riparins I, II and III in Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa. We analyzed the cytotoxicity by MTT assay in Vero cells and hemolytic action verified in human erythrocytes. The antimicrobial activity was determined by microdilution in broth against ATCC strains, identifying the susceptible species. Subsequently, only the MDR isolates of sensitive bacterial species were evaluated regarding its biofilm formation and ultrastructural changes. Riparin I presented low cytotoxicity and hemolytic percentage ranging from of 9.01%-12.97%. Only the riparin III that showed antimicrobial activity against MDR clinical isolates, and significant reduction in biofilm formation in S. aureus. Moreover, the riparin III promoted ultrastructural changes in bacterial cells, such as elongated cellular without bacterial septum, cells with a rugged appearance on the cell surface and cytoplasmic material extravasation. As has been noted riparin III has an inhibitory potential against biofilm formation in S. aureus, besides having antimicrobial activity and promoting ultrastructural changes in MDR clinical isolates. Thus, riparin III is an interesting alternative for further studies aiming to develop new therapeutic options.

Ultrastructural changes caused by the combination of intravenous immunoglobulin with meropenem, amikacin and colistin in multidrug-resistant Acinetobacter baumannii.

Acinetobacter baumannii is an opportunistic pathogen associated with increased morbidity and mortality in Healthcare-associated infections (HAI). Combination antimicrobial therapy, meropenem, amikacin and colistin, has been used as an alternative in multidrug-resistant (MDR) A. baumannii infections due to reduced treatment options. However, these combinations are not always effective and exhibit high toxicity. Empiric therapy of intravenous immunoglobulin (IVIG) associated with antimicrobials has shown promising results in bacterial infections, considering the immunomodulatory action of IVIG. Thus, the aim of this study was to determine the combined antimicrobial action and to describe the ultrastructural changes caused in ten MDR A. baumannii isolates submitted to IVIG alone and in combination with colistin, meropenem and amikacin. Minimum Inhibitory Concentration (MIC) of antimicrobials and checkerboard were determined. Isolates were submitted to 4 mg/mL of IVIG alone and in combination with different synergistic sub-MIC of antimicrobials tested, and processed for scanning electron microscopy. Nine bacterial isolates showed meropenem-resistant, two isolates had colistin-intermediate, and four isolates were considered intermediate to amikacin. Synergism in five isolates for meropenem/amikacin and meropenem/colistin were observed. Bacterial cells submitted to IVIG and meropenem, amikacin and colistin presented several ultrastructural changes, such as cell elongation and rupture, membrane roughness, incomplete cell division, cell surface "bubbles" and "depression". A. baumannii isolates presented high resistance to meropenem and synergism among evaluated antimicrobials. In addition, it was possible to verify in vitro that IVIG associated with meropenem, amikacin and colistin is a promising alternative for MDR A. baumannii infections. Thus, these data support the continued empirical use and stimulate in vivo analyzes with IVIG to search for new therapeutic options for HAI.

Perturbed neural activity disrupts cerebral angiogenesis during a postnatal critical period.

During the neonatal period, activity-dependent neural-circuit remodelling coincides with growth and refinement of the cerebral microvasculature. Whether neural activity also influences the patterning of the vascular bed is not known. Here we show in neonatal mice, that neither reduction of sensory input through whisker trimming nor moderately increased activity by environmental enrichment affects cortical microvascular development. Unexpectedly, chronic stimulation by repetitive sounds, whisker deflection or motor activity led to a near arrest of angiogenesis in barrel, auditory and motor cortices, respectively. Chemically induced seizures also caused robust reductions in microvascular density. However, altering neural activity in adult mice did not affect the vasculature. Histological analysis and time-lapse in vivo two-photon microscopy revealed that hyperactivity did not lead to cell death or pruning of existing vessels but rather to reduced endothelial proliferation and vessel sprouting. This anti-angiogenic effect was prevented by administration of the nitric oxide synthase (NOS) inhibitor L-NAME and in mice with neuronal and inducible NOS deficiency, suggesting that excessive nitric oxide released from hyperactive interneurons and glia inhibited vessel growth. Vascular deficits persisted long after cessation of hyperstimulation, providing evidence for a critical period after which proper microvascular patterning cannot be re-established. Reduced microvascular density diminished the ability of the brain to compensate for hypoxic challenges, leading to dendritic spine loss in regions distant from capillaries. Therefore, excessive sensorimotor stimulation and repetitive neural activation during early childhood may cause lifelong deficits in microvascular reserve, which could have important consequences for brain development, function and pathology.

"You Are Beautiful, No Matter What They Say": Applying An Evidence-Based Approach To Body Image Law.

Israeli and French Governments passed Body Image Laws that require models to have a minimum BMI or be of a healthy weight and if an image was modified to make the model appear thinner, it must have a warning. Are these laws merely symbolic, to focus a spotlight on this issue, or can they too have an impact? This article analyses some of the criticisms of the Body Image Laws by applying existing evidence from health research. Ultimately, it argues that there are many flaws with the Body Image Laws and that such a law should not be passed in Australia.

Green spaces are not all the same for the provision of air purification and climate regulation services: The case of urban parks.

The growing human population concentrated in urban areas lead to the increase of road traffic and artificial areas, consequently enhancing air pollution and urban heat island effects, among others. These environmental changes affect citizen's health, causing a high number of premature deaths, with considerable social and economic costs. Nature-based solutions are essential to ameliorate those impacts in urban areas. While the mere presence of urban green spaces is pointed as an overarching solution, the relative importance of specific vegetation structure, composition and management to improve the ecosystem services of air purification and climate regulation are overlooked. This avoids the establishment of optimized planning and management procedures for urban green spaces with high spatial resolution and detail. Our aim was to understand the relative contribution of vegetation structure, composition and management for the provision of ecosystem services of air purification and climate regulation in urban green spaces, in particular the case of urban parks. This work was done in a large urban park with different types of vegetation surrounded by urban areas. As indicators of microclimatic effects and of air pollution levels we selected different metrics: lichen diversity and pollutants accumulation in lichens. Among lichen diversity, functional traits related to nutrient and water requirements were used as surrogates of the capacity of vegetation to filter air pollution and to regulate climate, and provide air purification and climate regulation ecosystem services, respectively. This was also obtained with very high spatial resolution which allows detailed spatial planning for optimization of ecosystem services. We found that vegetation type characterized by a more complex structure (trees, shrubs and herbaceous layers) and by the absence of management (pruning, irrigation and fertilization) had a higher capacity to provide the ecosystems services of air purification and climate regulation. By contrast, lawns, which have a less complex structure and are highly managed, were associated to a lower capacity to provide these services. Tree plantations showed an intermediate effect between the other two types of vegetation. Thus, vegetation structure, composition and management are important to optimize green spaces capacity to purify air and regulate climate. Taking this into account green spaces can be managed at high spatial resolutions to optimize these ecosystem services in urban areas and contribute to improve human well-being.

Ecosystem services: Urban parks under a magnifying glass.

Urban areas' population has grown during the last century and it is expected that over 60% of the world population will live in cities by 2050. Urban parks provide several ecosystem services that are valuable to the well-being of city-dwellers and they are also considered a nature-based solution to tackle multiple environmental problems in cities. However, the type and amount of ecosystem services provided will vary with each park vegetation type, even within same the park. Our main goal was to quantify the trade-offs in ecosystem services associated to different vegetation types, using a spatially detailed approach. Rather than relying solely on general vegetation typologies, we took a more ecologically oriented approach, by explicitly considering different units of vegetation structure and composition. This was demonstrated in a large park (44ha) located in the city of Almada (Lisbon metropolitan area, Portugal), where six vegetation units were mapped in detail and six ecosystem services were evaluated: carbon sequestration, seed dispersal, erosion prevention, water purification, air purification and habitat quality. The results showed that, when looking at the park in detail, some ecosystem services varied greatly with vegetation type. Carbon sequestration was positively influenced by tree density, independently of species composition. Seed dispersal potential was higher in lawns, and mixed forest provided the highest amount of habitat quality. Air purification service was slightly higher in mixed forest, but was high in all vegetation types, probably due to low background pollution, and both water purification and erosion prevention were high in all vegetation types. Knowing the type, location, and amount of ecosystem services provided by each vegetation type can help to improve management options based on ecosystem services trade-offs and looking for win-win situations. The trade-offs are, for example, very clear for carbon: tree planting will boost carbon sequestration regardless of species, but may not be enough to increase habitat quality. Moreover, it may also negatively influence seed dispersal service. Informed practitioners can use this ecological knowledge to promote the role of urban parks as a nature-based solution to provide multiple ecosystem services, and ultimately improve the design and management of the green infrastructure. This will also improve the science of Ecosystem Services, acknowledging that the type of vegetation matters for the provision of ecosystem services and trade-offs analysis.

Body image diversity in the media: A content analysis of women's fashion magazines.

ISSUE ADDRESSED: The narrow representation of body image in the media has been linked to body dissatisfaction, particularly among readers of women's fashion magazines. Some countries have made efforts to improve body image diversity in the media and the fashion industry. This has included attempts to regulate minimum body size of models (eg, Israel, France), and the development of codes of practices such as the Australian Industry Code of Conduct on Body Image. However, there is little evidence of whether these efforts have impacted media content. METHOD: This study aimed to gauge the state of body image diversity in the print media 5 years after the introduction of the Australian Code of Conduct via a content analysis of 13 Australian women's fashion magazines published in 2015. RESULTS: Results revealed low levels of diversity in body size, ethnicity and age among models depicted in fashion magazine images. Models were predominantly young, white and underweight. CONCLUSION: The results suggest that efforts to improve body image diversity have had little impact on print media. Further research is needed to understand the barriers to increased diversity in the representation of body image in the media so that the industry and regulatory bodies can further address this important issue. This is increasingly pressing given the proliferation of content now enabled through online media platforms.

Evaluating green infrastructure in urban environments using a multi-taxa and functional diversity approach.

Forested areas within cities host a large number of species, responsible for many ecosystem services in urban areas. The biodiversity in these areas is influenced by human disturbances such as atmospheric pollution and urban heat island effect. To ameliorate the effects of these factors, an increase in urban green areas is often considered sufficient. However, this approach assumes that all types of green cover have the same importance for species. Our aim was to show that not all forested green areas are equal in importance for species, but that based on a multi-taxa and functional diversity approach it is possible to value green infrastructure in urban environments. After evaluating the diversity of lichens, butterflies and other-arthropods, birds and mammals in 31 Mediterranean urban forests in south-west Europe (Almada, Portugal), bird and lichen functional groups responsive to urbanization were found. A community shift (tolerant species replacing sensitive ones) along the urbanization gradient was found, and this must be considered when using these groups as indicators of the effect of urbanization. Bird and lichen functional groups were then analyzed together with the characteristics of the forests and their surroundings. Our results showed that, contrary to previous assumptions, vegetation density and more importantly the amount of urban areas around the forest (matrix), are more important for biodiversity than forest quantity alone. This indicated that not all types of forested green areas have the same importance for biodiversity. An index of forest functional diversity was then calculated for all sampled forests of the area. This could help decision-makers to improve the management of urban green infrastructures with the goal of increasing functionality and ultimately ecosystem services in urban areas.

Ultrastructural Changes in Clinical and Microbiota Isolates of Klebsiella pneumoniae Carriers of Genes bla SHV, bla TEM, bla CTX-M, or bla KPC When Subject to ß-Lactam Antibiotics.

The aim of this study was to characterize the ultrastructural effects caused by ß-lactam antibiotics in Klebsiella pneumoniae isolates. Three K. pneumoniae clinical isolates were selected for the study with resistance profiles for third-generation cephalosporins, aztreonam, and/or imipenem and with different resistance genes for extended-spectrum ß-lactamases (ESBL) or Klebsiella pneumoniae carbapenemase (KPC). Two K. pneumoniae isolates obtained from the microbiota, which were both resistant to amoxicillin and ampicillin, were also analyzed. In accordance with the susceptibility profile, the clinical isolates were subjected to subminimum inhibitory concentrations (sub-MICs) of cefotaxime, ceftazidime, aztreonam, and imipenem and the isolates from the microbiota to ampicillin and amoxicillin, for analysis by means of scanning and transmission electron microscopy. The K. pneumoniae isolates showed different morphological and ultrastructural changes after subjection to ß-lactams tested at different concentrations, such as cell filamentation, loss of cytoplasmic material, and deformation of dividing septa. Our results demonstrate that K. pneumoniae isolates harboring different genes that encode for ß-lactamases show cell alterations when subjected to different ß-lactam antibiotics, thus suggesting that they possess residual activity in vitro, despite the phenotypic resistance presented in the isolates analyzed.

Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation.

Angiogenesis, the growth of new blood vessels from pre-existing vasculature, is a key process in several pathological conditions, including tumour growth and age-related macular degeneration. Vascular endothelial growth factors (VEGFs) stimulate angiogenesis and lymphangiogenesis by activating VEGF receptor (VEGFR) tyrosine kinases in endothelial cells. VEGFR-3 (also known as FLT-4) is present in all endothelia during development, and in the adult it becomes restricted to the lymphatic endothelium. However, VEGFR-3 is upregulated in the microvasculature of tumours and wounds. Here we demonstrate that VEGFR-3 is highly expressed in angiogenic sprouts, and genetic targeting of VEGFR-3 or blocking of VEGFR-3 signalling with monoclonal antibodies results in decreased sprouting, vascular density, vessel branching and endothelial cell proliferation in mouse angiogenesis models. Stimulation of VEGFR-3 augmented VEGF-induced angiogenesis and sustained angiogenesis even in the presence of VEGFR-2 (also known as KDR or FLK-1) inhibitors, whereas antibodies against VEGFR-3 and VEGFR-2 in combination resulted in additive inhibition of angiogenesis and tumour growth. Furthermore, genetic or pharmacological disruption of the Notch signalling pathway led to widespread endothelial VEGFR-3 expression and excessive sprouting, which was inhibited by blocking VEGFR-3 signals. Our results implicate VEGFR-3 as a regulator of vascular network formation. Targeting VEGFR-3 may provide additional efficacy for anti-angiogenic therapies, especially towards vessels that are resistant to VEGF or VEGFR-2 inhibitors.

Time to Start a New Enhanced Recovery After Surgery (ERAS): A Retrospective Cohort Study.

BACKGROUND: The enhanced recovery after surgery (ERAS®) is a multimodal perioperative care pathway designed to reduce surgical stress and ultimately improve patient recovery and outcome. It can require significant resources but with proven benefits. The main goal of this study was to perform a diagnostic assessment of perioperative practice in a local colorectal surgical center. METHODS: 93 patients who underwent elective colorectal surgery from January to December 2022 were analyzed. Preadmission, preoperative, and postoperative data of all patients were collected in a database developed by the researchers, according to ERAS® guidelines. Descriptive statistics were employed to summarize demographic and clinical characteristics. Chi-square and T-test were performed to identify possible associations between categorical variables and postoperative complications. RESULTS: Overall analysis showed deficient preoperative patient optimization, especially regarding nutritional counseling and supplementation, smoking and alcohol cessation, anemia treatment (9%), and pre-anesthetic medication (42%). Removal of invasive devices was significantly delayed (removal of urinary catheter average on the fourthday and surgical drain average on the fifth day) in the postoperatively period and oral intake (average onset on the sixth day). Both contribute to hospital length of stay (mean of 13 days) and a significant number of complications. CONCLUSION: The results lead us to an individual and multidisciplinary reflection on current practices and outcomes. ERAS® program, already adopted by many centers, could have a positive impact on the immediate postoperative recovery of colorectal patients in Funchal Central Hospital and implementation seems necessary.

Hyperbaric Medicine in Pediatrics - reality of a Portuguese reference center.

OBJECTIVE: To identify and characterize the population of Pediatric patients referred to our hyperbaric oxygen therapy center. METHODS: Retrospective and observational study, including pediatric patients treated with hyperbaric oxygen therapy, from 2006 to 2021, at the hyperbaric medicine reference center in the north of Portugal. Variables of interest were extracted from electronic medical records. RESULTS: Our study included 134 patients. The most frequent reasons for referral were carbon monoxide poisoning (n=59) and sudden sensorineural hearing loss (n=41). In 75 cases (56%), treatment was initiated in an urgent context. Symptom presentation at Emergency Department varied among patients, the most frequent being headache and nausea/vomiting. Concerning carbon monoxide poisoning, the most common sources were water heater, fireplace/brazier, and boiler. Regarding adverse effects, it was identified one case of intoxication by oxygen and four cases of middle ear barotrauma. CONCLUSIONS: The most frequent cause for referral was carbon monoxide poisoning. All patients evolved favorably, with few side effects being reported, emphasizing the safety of this therapy. While most pediatricians may not be aware of the potential benefits arising with hyperbaric oxygen therapy, it is of upmost importance to promote them, so that this technique is increasingly implemented.

Effects of the COVID-19 Pandemic on Breastfeeding Initiation and Duration: A Retrospective Cohort Study.

INTRODUCTION AND OBJECTIVES: Several studies support the health benefits of breastfeeding for both the mother and the newborn. However, a significant number of mothers discontinue breastfeeding within the first six months of childbirth, with several factors influencing breastfeeding adherence. The purpose of this study is to assess the impact of the COVID-19 pandemic on the prevention of mother-to-newborn infection transmission, breastfeeding patterns and duration, and the incidence of other infections during the first year of life. METHODS: Data from a sample of 39 mothers who gave birth at the Hospital Pedro Hispano in Porto, Portugual, between March 2020 and November 2021 were collected and a telephone questionnaire was administered. Statistical analysis was conducted using R software, v. 4.2.1 (R Foundation for Statistical Computing, Vienna, Austria). RESULTS AND DISCUSSION: In terms of the impact of the COVID-19 norm 18/2020, which went into effect on March 30th, our research found that the type of feeding during hospitalization was significantly influenced by this norm (X2=10.30, p=0.006). We also confirmed that mothers who received home assistance breastfed for an extra 4.5 months (95% CI: 1-7.5) compared with mothers who did not receive such assistance. Regarding the effect of COVID-19 and breastfeeding on newborn health, our study found that if the total duration of breastfeeding is less than six months, an infection is approximately five times more likely (95% CI = 1.06- 29.56). CONCLUSION: Overall, the findings of this study indicate that the efforts implemented at Hospital Pedro Hispano to limit the effects of the COVID-19 pandemic had some effect on immediate breastfeeding patterns, but not on the total duration of breastfeeding or newborn health. Nonetheless, more continuous assistance at home would have been beneficial.

Laboratory Assessment of the Effects of AGA@4life Multidisciplinary Intervention on the Inflammatory Profile, MMPs, and TIMPs in a Geriatric Population.

In recent years, the world's aging population has increased, contributing to the development of age-related pathologies, which have been aggravated by physical inactivity and excessive fat intake. This study aimed to evaluate the effect of implementing a nutritional program (control group-CG) combined with exercise (intervention group-IG) on the inflammatory profile, MMPs, and TIMPs in a group of 34 elderly participants (IG, n = 18; CG, n = 16). Participants underwent a full multidisciplinary diagnostic evaluation (T0), with the gathering of clinical information and biochemical and hematological determinations being re-evaluated eight weeks later (T1). A diet manual was made, which provided a selection of different types of diets resulting from the nutritional needs of the different users at the center. The aerobic exercise consisted of two sessions per week with a total duration of 1 h. The laboratory evaluation was performed by slot blot. Statistical analysis included a paired sample t-test and Spearman's correlation coefficient. We observed that in the IG, there was a significant increase at T1 of TNF-α (p < 0.05) and MMP-2 (p < 0.05), without changes in IL-6 and MMP-9, showing that the intervention did not cause an exacerbated inflammatory response in exercised elderly people. The intervention program implemented showed potential to contribute to better active aging strategies, taking advantage of the known benefits of exercise without inducing a harmful inflammatory response in elderly participants.

Linear Immunoglobulin A Dermatosis: A Rare Case Illustrating Successful Treatment With Dapsone.

This is the case report of a previously healthy four-year-old girl with a history of upper airway infection that was treated with a ß-lactam antibiotic. She was seen in the emergency department one month later with vesiculobullous lesions with clear content that were isolated or grouped in rosettes. Direct immunofluorescence showed baseline linear positivity for immunoglobulin A (IgA) (+) and fibrinogen-positive bullous content with absent remaining immunosera expression. The observed results were compatible with linear IgA bullous dermatosis. After confirming the diagnosis and excluding glucose-6-phosphate dehydrogenase (G6PD) deficiency, dapsone was added to the initial treatment with systemic and topical corticosteroids. This case report is a reminder of the importance of a high index of clinical suspicion for this condition to reach a timely diagnosis.

In vitro evaluation of human intravenous immunoglobulin in combination with antimicrobials and human serum against multidrug-resistant isolates of Acinetobacter baumannii.

The high incidence of multidrug-resistant (MDR) Acinetobacter baumannii has been a challenge for health worldwide, due to the reduction of therapeutic options, making the use of antimicrobial combinations necessary for the treatment, such as meropenem, amikacin, and colistin. Antibodies against bacterial species, mainly immunoglobulins G (IgG), are produced for acting as effector mechanisms (neutralization, opsonization, phagocytosis, and complement system activation). Some studies have demonstrated promising results of IgG in combination with antimicrobial preparations against bacterial infections, in which the direct action of IgG has restored the immune system balance. Serious problem caused by the increase of MDR A. baumannii isolates results in a constant search for therapeutic alternatives to defeat these infections. However, this study aims to verify in vitro the phagocytosis rate of the A. baumannii-infected human monocytes, as well as to analyze possible morphological changes induced by intravenous immunoglobulin G (IVIG) with human serum in association with antimicrobials. The phagocytosis rate and bacterial cell binding capacity of IVIG were determined for two A. baumannii isolates submitted to 4 mg/mL of human IVIG alone and in combination with different sub-minimum inhibitory concentrations (sub-MICs) of meropenem, amikacin, and colistin and processed for indirect immunofluorescence. Subsequently, these isolates were resubmitted and coupled with human serum and processed for scanning electron microscopy. There was no statistical difference for phagocytosis rates in the isolates tested. Bacterial isolates showed alterations in cell morphology when exposed to IVIG/human serum alone and in combination with antimicrobials such as alteration in shape, wrinkling, membrane depression, and especially cell rupture with extravasation of cytoplasmic material. The isolates visually differed in the IVIG binding to the bacterial cell, with higher fluorescence intensity, which corresponds to the highest IVIG binding, in the isolate more sensitive to meropenem, amikacin, and colistin. No differences between treatments were observed in the IVIG binding to the bacterial cell. The combined action of IVIG with meropenem, amikacin, and colistin against A. baumannii MDR isolates induced several bacterial cell damages. And when associated with human serum, a massive destruction of cells can be observed. These results may suggest the analysis of the use of IgG preparations for the treatment of A. baumannii MDR infections.

Treatment of auditory verbal hallucinations with transcranial magnetic stimulation in a patient with psychotic major depression: one-year follow-up.

Auditory verbal hallucinations (AVH) in patients with schizophrenia can respond to repetitive transcranial magnetic stimulation (TMS). We report the therapeutic utility of rTMS in a 48-year-old patient with a 20-year history of severe depression (five suicidal gestures and previous failure of ECT) and internal AVH. First, 20 Hz rTMS to the left prefrontal cortex for 3 weeks significantly improved depression (BDI-II: 89% change, lasting 14 months along with weekly/bi-weekly maintenance treatments), but AVH remained unchanged. The patient also underwent a further course of the left temporo-parietal 1 Hz rTMS and amelioration of AVH severity was achieved (PSYRATS-AH: 53% change) and maintained at 1-year follow-up. AVH respond to rTMS in disorders other than schizophrenia. Furthermore, targeted rTMS to different brain regions can address diverse symptoms in neuropsychiatric conditions.

Polymerized Laminin-521: A Feasible Substrate for Expanding Induced Pluripotent Stem Cells at a Low Protein Concentration.

Laminins (LNs) play a central role in the self-assembly and maintenance of basement membranes and are involved in critical interactions between cells and other extracellular matrix (ECM) proteins. Among the defined, xeno-free ECM culture matrices, LNs-namely LN521-have emerged as promising coating systems for the large-scale expansion of induced pluripotent stem cells (iPSCs). The biologic activity of LNs is enhanced by their acidification-induced self-polymerization into a cell-associated network called polylaminin (polyLN), which can recapitulate the native-like polymeric array in a cell-free system. Here, we show for the first time to our knowledge that polyLN521 displays a native-like hexagonal-like structure and that, at basal and low concentrations, it permits the large-scale expansion of human iPSCs. Human iPSCs expanded with polyLN521 maintained the pluripotent state and showed no impairment of karyotype stability or telomere length. These results suggest that low-concentration polyLN521 is a stable and cost-effective coating for large-scale iPSC expansion.

Modulatory effects of theta burst stimulation on cerebellar nonsomatic functions.

Clinical and functional imaging studies suggest that the cerebellar vermis is involved in the regulation of a range of nonsomatic functions including cardiovascular control, thirst, feeding behavior, and primal emotions. Cerebello-hypothalamic circuits have been postulated to be a potential neuroanatomical substrate underlying this modulation. We tested this putative relationship between the cerebellar vermis and nonsomatic functions by stimulating the cerebellum noninvasively via neuronavigated transcranial magnetic stimulation. In this randomized, counter-balanced, within-subject study, intermittent theta burst stimulation (TBS) was applied on three different days to the vermis and the right and left cerebellar hemispheres of 12 right-handed normal subjects with the aim of modulating activity in the targeted cerebellar structure. TBS-associated changes were investigated via cardiovascular monitoring, a series of emotionally arousing picture stimuli, subjective analog scales for primal emotions, and the Profile of Mood States test. All 36 sessions of cerebellar stimulation were tolerated well without serious adverse events. Cardiovascular monitoring pointed to a mild but significant decrease in heart rate subsequent to vermal stimulation; no changes were detected in systolic or diastolic blood pressure measurements. Subjective ratings detected a significant increase in Thirst and a trend toward increased Appetite following vermal stimulation. These observations are consistent with existing neurophysiological and neuroimaging data indicating a role for the cerebellum in the regulation of visceral responses. In conjunction with the modulatory function of the cerebellum, our results suggest a role for the vermis in somatovisceral integration likely through cerebello-hypothalamic pathways. Further research is warranted to elucidate the potential mechanisms underlying the cerebellar modulation of nonsomatic functions.

Guidance of vascular development: lessons from the nervous system.

The vascular system of vertebrates consists of an organized, branched network of arteries, veins, and capillaries that penetrates all the tissues of the body. One of the most striking features of the vascular system is that its branching pattern is highly stereotyped, with major and secondary branches forming at specific sites and developing highly conserved organ-specific vascular patterns. The factors controlling vascular patterning are not yet completely understood. Recent studies have highlighted the anatomic and structural similarities between blood vessels and nerves. The 2 networks are often aligned, with nerve fibers and blood vessels following parallel routes. Furthermore, both systems require precise control over their guidance and growth. Several molecules with attractive and repulsive properties have been found to modulate the proper guidance of both nerves and blood vessels. These include the Semaphorins, the Slits, and the Netrins and their receptors. In this review, we describe the molecular mechanisms by which blood vessels and axons achieve proper path finding and the molecular cues that are involved in their guidance.