Intrathecal AAVrh10 corrects biochemical and histological hallmarks of mucopolysaccharidosis VII mice and improves behavior and survival.

Mucopolysaccharidosis (MPS) type VII is a lysosomal storage disease caused by ß-glucuronidase deficiency, prompting glycosaminoglycan accumulation in enlarged vesicles, leading to peripheral and neuronal dysfunction. Here, we present a gene therapy strategy using lumbar puncture of AAVrh10 encoding human ß-glucuronidase (AAVrh10-GUSB) to adult MPS VII mice. This minimally invasive technique efficiently delivers the recombinant vector to the cerebrospinal fluid (CSF) with a single intrathecal injection. We show that AAVrh10 delivery to the CSF allows global, stable transduction of CNS structures. In addition, drainage of AAVrh10-GUSB from the CSF to the bloodstream resulted in the transduction of somatic organs such as liver, which provided a systemic ß-glucuronidase source sufficient to achieve serum enzyme activity comparable to wild type mice. ß-glucuronidase levels were enough to correct biochemical and histopathological hallmarks of the disease in the CNS and somatic organs at short and long term. Moreover, the progression of the bone pathology was also reduced. Importantly, the biochemical correction led to a significant improvement in the physical, cognitive and emotional characteristics of MPS VII mice, and doubling their life span. Our strategy may have implications for gene therapy in patients with lysosomal storage diseases.

Reprogramming human B cells into induced pluripotent stem cells and its enhancement by C/EBPα.

B cells have been shown to be refractory to reprogramming and B-cell-derived induced pluripotent stem cells (iPSC) have only been generated from murine B cells engineered to carry doxycycline-inducible Oct4, Sox2, Klf4 and Myc (OSKM) cassette in every tissue and from EBV/SV40LT-immortalized lymphoblastoid cell lines. Here, we show for the first time that freshly isolated non-cultured human cord blood (CB)- and peripheral blood (PB)-derived CD19+CD20+ B cells can be reprogrammed to iPSCs carrying complete VDJH immunoglobulin (Ig) gene monoclonal rearrangements using non-integrative tetracistronic, but not monocistronic, OSKM-expressing Sendai Virus. Co-expression of C/EBPα with OSKM facilitates iPSC generation from both CB- and PB-derived B cells. We also demonstrate that myeloid cells are much easier to reprogram than B and T lymphocytes. Differentiation potential back into the cell type of their origin of B-cell-, T-cell-, myeloid- and fibroblast-iPSCs is not skewed, suggesting that their differentiation does not seem influenced by 'epigenetic memory'. Our data reflect the actual cell-autonomous reprogramming capacity of human primary B cells because biased reprogramming was avoided by using freshly isolated primary cells, not exposed to cytokine cocktails favoring proliferation, differentiation or survival. The ability to reprogram CB/PB-derived primary human B cells offers an unprecedented opportunity for studying developmental B lymphopoiesis and modeling B-cell malignancies.

[Perception of UCI nurses in relation with satisfactory care: convergences and divergences with the perception of critical patients]. / Percepción de las enfermeras de UCI en relación al cuidado satisfactorio: convergencias y divergencias con la percepción del paciente crítico.

OBJECTIVE: Explore convergences and divergences between perception of nurses and of critically ill patients, in relation to the satisfactory care given and received. METHODS: It is part of a larger qualitative study, according to the Grounded Theory. Carried out in 3 intensive care units with 34 boxes. Sampling theoretical profiles with n=19 patients and n=7 nurses after data saturation. Recruitment of patients included in the profiles of elderly and long-stay got stretched over some time due to the low incidence of cases. Data collection consisted of: in-depth interview to critically ill patients, group discussion of expert nurses in the critical care patient and field diary. Analysis themed on Grounded Theory according Strauss and Corbin: open coding, axial and selective. Analysis followed criteria of Guba and Lincoln rigor, Calderón quality and Gastaldo and McKeever ethical reflexivity. There was a favorable report from the ethical committee of the Hospital and informed consent of the participants. RESULTS: Four matching categories were found: professional skills, human, technical and continued care. Combination of these elements creates feelings of security, calmness and feeling like a person, allowing the patient a close and trusting relationship with the nurse who takes individualized care. Not divergent categories were found. CONCLUSIONS: Perceptions of nurses in relation to care match perceptions of critically ill patients in both the definition and dimensions upon satisfactory care.

Experimental diabetes in neonatal mice induces early peripheral sensorimotor neuropathy.

Animal models of diabetes do not reach the severity of human diabetic neuropathy but relatively mild neurophysiological deficits and minor morphometric changes. The lack of degenerative neuropathy in diabetic rodent models seems to be a consequence of the shorter length of the axons or the shorter animal life span. Diabetes-induced demyelination needs many weeks or even months before it can be evident by morphometrical analysis. In mice myelination of the peripheral nervous system starts at the prenatal period and it is complete several days after birth. Here we induced experimental diabetes to neonatal mice and we evaluated its effect on the peripheral nerve 4 and 8 weeks after diabetes induction. Neurophysiological values showed a decline in sensory nerve conduction velocity at both time-points. Morphometrical analysis of the tibial nerve demonstrated a decrease in the number of myelinated fibers, fiber size and myelin thickness at both time-points studied. Moreover, aldose reductase and poly(ADP-ribose) polymerase activities were increased even if the amount of the enzyme was not affected. Thus, type 1 diabetes in newborn mice induces early peripheral neuropathy and may be a good model to assay pharmacological or gene therapy strategies to treat diabetic neuropathy.

Factors associated with migration in individuals affected by leprosy, maranhão, Brazil: an exploratory cross-sectional study.

In Brazil, leprosy is endemic and concentrated in high-risk clusters. Internal migration is common in the country and may influence leprosy transmission and hamper control efforts. We performed a cross-sectional study with two separate analyses evaluating factors associated with migration in Brazil's Northeast: one among individuals newly diagnosed with leprosy and the other among a clinically unapparent population with no symptoms of leprosy for comparison. We included 394 individuals newly diagnosed with leprosy and 391 from the clinically unapparent population. Of those with leprosy, 258 (65.5%) were birth migrants, 105 (26.6%) were past five-year migrants, and 43 (10.9%) were circular migrants. In multivariate logistic regression, three independent factors were found to be significantly associated with migration among those with leprosy: (1) alcohol consumption, (2) separation from family/friends, and (3) difficulty reaching the healthcare facility. Separation from family/friends was also associated with migration in the clinically unapparent population. The health sector may consider adapting services to meet the needs of migrating populations. Future research is needed to explore risks associated with leprosy susceptibility from life stressors, such as separation from family and friends, access to healthcare facilities, and alcohol consumption to establish causal relationships.

[Review of the knowledge on acute kidney failure in the critical patient]. / Revisión de conocimientos sobre el fracaso renal agudo en el contexto del paciente crítico.

Acute renal failure affects from 1% to 25% of patients admitted to intensive care units. These figures vary depending on the population studied and criteria. The complications of acute renal failure (fluid overload, metabolic acidosis, hyperkalemia, bleeding) are treated. However, mortality remains high despite the technological advances of recent years because acute renal failure is usually associated with sepsis, respiratory failure, serious injury, surgical complications or consumption coagulopathy. Mortality ranges from 30% to 90%. Although there is no universally accepted definition, the RIFLE classification gives us an operational tool to define the degree of acute renal failure and to standardize the initiation of renal replacement techniques as well as to evaluate the results. Therefore, nurses working within the intensive care unit must be familiar with this disease, with its treatment (drug or alternative) and with the prevention of possible complications. Equally, they must be capable of detecting the manifestations of dependency each one of the basic needs and to be able to identify the collaboration problems in order to achieve an individualized care plan.

[Update in continuous renal replacement techniques]. / Actualización en técnicas continuas de reemplazo renal.

Acute renal failure affects 25% of patients hospitalized in intensive care units. Despite technological advances, the mortality of these patients is still high due to its associated complications. Continuous renal replacement techniques are one of the treatments for acute renal failure because they make it possible to treat the complications and decrease mortality. The nurse's knowledge and skills regarding these techniques will be decisive for the success of the therapy. Consequently, the nurse's experience and training are key components. The objective of this article is to update the knowledge on continuous renal replacement techniques. Keeping this in mind, a review has been made of the physical and chemical principles such as diffusion and convection, among others. A description of the different continuous renal replacement techniques, a presentation of the main vascular access, and a description of the nursing cares and complications related to techniques used have also been provided.

Investigation of a scabies outbreak in a kindergarten in Constance, Germany.

In industrialized countries, scabies occurs sporadically or in the form of protracted epidemics, typically in nursing homes for elderly people. Outbreaks of scabies in a kindergarten are very rare. The main goal of our study was to investigate an outbreak of scabies in a kindergarten and to identify risk factors for the infestation with the ectoparasitosis. We investigated an outbreak of scabies in a kindergarten in the City of Constance, southern Germany, with a particular pedagogical concept. Risk factors indicating a transmission of Sarcoptes mites through body contact or via fomites were assessed using questionnaires and by following the daily routine in the kindergarten. A total of 16 cases were identified. The attack rate was significantly higher in nursery teachers (risk ratio 42.1) compared to children (risk ratio 10.5). In all cases, scabies had developed rather recently, with minimal clinical manifestations. In nursery teachers, the probability of scabies was 4.4 times higher in those teachers who hugged children regularly. Children who preferably played with their own soft toys had a lower probability of developing scabies [risk ratio 0.14, 95 % confidence interval (CI) 0.05-0.42; p = 0.04]. It seems conceivable that the particular pedagogical concept of the kindergarten favored the spread of Sarcoptes mites. We were unable to show whether transmission had preferably occurred through body contact or via fomites.

[Perception of the critical patient on nursing cares: an approach to the concept of satisfaction]. / La percepción del paciente crítico sobre los cuidados enfermeros: una aproximación al concepto de satisfacción.

INTRODUCTION: Level of satisfaction is a key indicator of quality of care. There are many tools that measure satisfaction with nursing care, however they do not respond to the reality of the critical care patient or to our context. OBJECTIVES: To define and to identify the dimensions of the satisfaction of patients admitted to the intensive care unit of a tertiary hospital with nursing cares and to define and identify the dimensions of the concept of satisfaction from their point of view. MATERIAL AND METHODS: A qualitative research study was conducted according to the Grounded Theory Method in three Intensive Care Units with 34 individual boxes, with theoretical sampling. Nineteen patients remained after data saturation sampling. Data collection was obtained through recorded in-depth interviews and field logbook. Contents analysis was made according to the Grounded Theory. Guba and Lincoln rigor's criteria were followed. There was a favorable report from the Hospital's Ethics Committee and informed consent was obtained from the patients. RESULTS: Four categories were found: The definition and dimensions of the satisfaction concept, expectations and life experiences. The participants included the following dimensions in their satisfaction definition: professional competences, human, technical and continuous cares. The combination of these elements produces feelings of security, calmness, being monitored, feeling like a person, perceiving a close relationship and trustfulness with the nurse who performs the individualized cares. CONCLUSIONS: The definition and dimensions of satisfaction concept from the patient's point of view show the important aspects of the person and also clarify their dimensions, allowing the construction of tools more in line with the context and real perception.

Schwann cell targeting via intrasciatic injection of AAV8 as gene therapy strategy for peripheral nerve regeneration.

Efficient transduction of the peripheral nervous system (PNS) is required for gene therapy of acquired and inherited neuropathies, neuromuscular diseases and for pain treatment. We have characterized the tropism and transduction efficiency of different adeno-associated vectors (AAV) pseudotypes after sciatic nerve injection in the mouse. Among the pseudotypes tested, AAV2/1 transduced both Schwann cells and neurons, AAV2/2 infected only sensory neurons and AAV2/8 preferentially transduced Schwann cells. AAV2/8 expression in the sciatic nerve was detected up to 10 weeks after administration, the latest time point analyzed. The injected mice developed neutralizing antibodies against all AAVs tested; the titers were higher against AAV2/1 than AAV2/2 and were the lowest for AAV2/8, correlating with a higher transgene expression overtime. AAV2/8 coding for ciliary neurotrophic factor (CNTF) led to an upregulation of P0 and PMP22 myelin proteins, four weeks after transduction of injured sciatic nerves. Importantly, CNTF-transduced mice showed a significant increase in both GAP43 expression in sensory neurons, a marker of axonal regeneration, and the compound muscle action potential. These results prove the utility of AAV8 as a gene therapy vector for Schwann cells to treat myelin disorders or to improve nerve regeneration.

A simple method for rapid community assessment of tungiasis.

OBJECTIVE: To evaluate a rapid assessment method to estimate the overall prevalence of tungiasis and severity of disease in endemic communities. METHODS: We analysed data from 10 population-based surveys on tungiasis, performed in five endemic communities in Brazil and Nigeria between 2001 and 2008. To assess the association between occurrence of tungiasis on six defined topographic areas of the feet and the true prevalence/prevalence of severe disease, linear regression analyses were performed. Estimated prevalences were calculated for each of the 10 surveys and compared to true prevalences. We then selected the most useful topographic localization to define a rapid assessment method, based on the strength of association and operational aspects. RESULTS: In total, 7121 individuals of the five communities were examined. Prevalence of tungiasis varied between 21.1% and 54.4%. The presence of periungual lesions on the toes was identified as the most useful rapid assessment to estimate the prevalence of tungiasis (absolute errors: -4% to +3.6%; R(2 )=96%; P < 0.0001). Prevalence of severe tungiasis (>20 lesions) was also estimated by the method (absolute errors: -3.1% to +2.5%; R(2 )=76%; P = 0.001). CONCLUSION: Prevalence of tungiasis and prevalence of severe disease can be reliably estimated in communities with distinct cultural and geographical characteristics, by applying a simple and rapid epidemiological method. This approach will help to detect high-risk communities and to monitor control measures aimed at the reduction of tungiasis.

Prevalence and risk factors of hookworm-related cutaneous larva migrans in a rural community in Brazil.

The epidemiology of hookworm-related cutaneous larva migrans (HrCLM) in endemic communities is not well understood. To describe the prevalence of HrCLM and to identify environmental and behavioural risk factors for the infestation, two cross-sectional surveys were conducted in a small village in an endemic area of north-eastern Brazil - one in the rainy season and the other in the dry season. The members of each household were examined clinically for the presence of HrCLM, and information on possible risk factors was collected. HrCLM, which was diagnosed in 45 individuals in the rainy-season survey and in 17 in the dry-season survey, was significantly more prevalent in the rainy season (4.4% v. 1.7%; P<0.001). The age-specific prevalences peaked, at 14.9%, in infants and children aged < or = 4 years. In a logistic regression analysis, the independent risk factors for current infestation or infestation in the preceding 6 months were identified as young age (odds ratio = 0.96; 95% confidence interval = 0.94-0.98), living in a house without a solid floor (odds ratio = 1.99; 95% confidence interval = 1.22-3.23), and walking barefoot (odds ratio = 1.77; 95% confidence interval = 1.12-2.80). In the study area, therefore, HrCLM is a common parasitic skin disease in children, is associated with behavioural and environmental risk factors, and shows marked seasonality in its prevalence. Local control of HrCLM should be based primarily on the health education of mothers and the elder girls who take care of their younger siblings.