Digital PCR methods improve detection sensitivity and measurement precision of low abundance mtDNA deletions

Mitochondrial DNA (mtDNA) mutations are a common cause of primary mitochondrial disorders, and have also been implicated in a broad collection of conditions, including aging, neurodegeneration, and cancer. Prevalent among these pathogenic variants are mtDNA deletions, which show a strong bias for the loss of sequence in the major arc between, but not including, the heavy and light strand origins of replication. Because individual mtDNA deletions can accumulate focally, occur with multiple mixed breakpoints, and in the presence of normal mtDNA sequences, methods that detect broad-spectrum mutations with enhanced sensitivity and limited costs have both research and clinical applications. In this study, we evaluated semi-quantitative and digital PCR-based methods of mtDNA deletion detection using double-stranded reference templates or biological samples. Our aim was to describe key experimental assay parameters that will enable the analysis of low levels or small differences in mtDNA deletion load during disease progression, with limited false-positive detection. We determined that the digital PCR method significantly improved mtDNA deletion detection sensitivity through absolute quantitation, improved precision and reduced assay standard error

Understanding lived experiences of food insecurity through a paraliminality lens

This article examines lived experiences of food insecurity in the United Kingdom as a liminal phenomenon. Our research is set within the context of austerity measures, welfare reform and the precarity experienced by increasing numbers of individuals. Drawing on original qualitative data, we highlight diverse food insecurity experiences as transitional, oscillating between phases of everyday food access to requiring supplementary food, which are both empowering and reinforcing of food insecurity. We make three original contributions to existing research on food insecurity. First, we expand the scope of empirical research by conceptualising food insecurity as liminal. Second, we illuminate shared social processes and practices that intersect individual agency and structure, co-constructing people’s experiences of food insecurity. Third, we extend liminality theory by conceptualising paraliminality, a hybrid of liminal and liminoid phenomena that co-generates a persistent liminal state. Finally, we highlight policy implications that go beyond short-term emergency food access measures

Assessment of optimal strategies in a two-patch dengue transmission model with seasonality

Emerging and re-emerging dengue fever has posed serious problems to public health officials in many tropical and subtropical countries. Continuous traveling in seasonally varying areas makes it more difficult to control the spread of dengue fever. In this work, we consider a two-patch dengue model that can capture the movement of host individuals between and within patches using a residence-time matrix. A previous two-patch dengue model without seasonality is extended by adding host demographics and seasonal forcing in the transmission rates. We investigate the effects of human movement and seasonality on the two-patch dengue transmission dynamics. Motivated by the recent Peruvian dengue data in jungle/rural areas and coast/urban areas, our model mimics the seasonal patterns of dengue outbreaks in two patches. The roles of seasonality and residence-time configurations are highlighted in terms of the seasonal reproduction number and cumulative incidence. Moreover, optimal control theory is employed to identify and evaluate patch-specific control measures aimed at reducing dengue prevalence in the presence of seasonality. Our findings demonstrate that optimal patch-specific control strategies are sensitive to seasonality and residence-time scenarios. Targeting only the jungle (or endemic) is as effective as controlling both patches under weak coupling or symmetric mobility. However, focusing on intervention for the city (or high density areas) turns out to be optimal when two patches are strongly coupled with asymmetric mobility.ope

The characteristics of the mechanoreceptors of the hip with arthrosis

Mechanoreceptors have been extensively studied in different joints and distinct signals that convey proprioceptive information to the cortex. Several clinical reports have established a link between the number of mechanoreceptors and a deficient proprioceptive system; however, little or no literature suggest concentration of mechanoreceptors might be affected by hip arthrosis. The purpose of this study is first to determine the existence of mechanoreceptors and free nerve endings in the hip joint and to distinguish between their conditions: those with arthrosis and without arthrosis. Samples of 45 male hips were analyzed: 30 taken from patients with arthrosis that were submitted to total arthroplasty and 15 taken from male cadavers without arthrosis. The patients' ages ranged from 38 to75 years (average 56.5) and the cadavers' ages ranged from 21 to 50 years (average 35.5). The capsule, labrum, and femoral head ligament tissues were obtained during the arthroplasty procedure from 30 patients with arthrosis and from 15 male cadavers. The tissue was cut into fragments of around 3 mm. Each fragment was then immediately stained with gold chloride 1% solution and divided into sections of 6 μm thickness. The Mann-Whitney test was used for two groups and the ANOVA, Friedman and Kruskal-Wallis tests for more than two groups. Results show the mechanoreceptors (Pacini, Ruffini and Golgi corpuscles) and free nerve endings are present in the capsule, femoral head ligament, and labrum of the hip joint. When all the densities of the nerve endings were examined with regard to those with arthrosis and those without arthrosis, the mechanoreceptors of cadavers without arthrosis were found to be more pronounced and an increase in free nerve endings could be observed (p = 0.0082). Further studies, especially electrophysiological studies, need to be carried out to clarify the functions of the mechanoreceptors in the joints

Enzymatic Solid-Phase Reactor Based on Silica Organofunctionalized with p-Phenylenediamine for Electrochemical Detection of Phenolic Compounds

A new biomaterial, based on silica organofunctionalized with p-phenylenediamine (p-PDA) and the enzyme peroxidase, was used in the development of an enzymatic solid-phase reactor. The analytical techniques used in the characterization showed that the organic ligand was incorporated into the silica matrix. Thus, the silica modified with p-PDA allowed the incorporation of peroxidase by the electrostatic interaction between the carboxylic groups present in the enzyme molecules and the amino groups attached to the silica. The enzymatic solid-phase reactor was used for chemical oxidation of phenols in 1 4-benzoquinone that was then detected by chronoamperometry. The system allowed the analysis of hydroquinone with a detection limit of 83.6 nmol L −1 . Thus, the new material has potential in the determination of phenolic compounds river water samples

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

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In Vivo Approaches Reveal a Key Role for DCs in CD4+ T Cell Activation and Parasite Clearance during the Acute Phase of Experimental Blood-Stage Malaria

Dendritic cells (DCs) are phagocytes that are highly specialized for antigen presentation. Heterogeneous populations of macrophages and DCs form a phagocyte network inside the red pulp (RP) of the spleen, which is a major site for the control of blood-borne infections such as malaria. However, the dynamics of splenic DCs during Plasmodium infections are poorly understood, limiting our knowledge regarding their protective role in malaria. Here, we used in vivo experimental approaches that enabled us to deplete or visualize DCs in order to clarify these issues. To elucidate the roles of DCs and marginal zone macrophages in the protection against blood-stage malaria, we infected DTx (diphtheria toxin)-treated C57BL/6.CD11c-DTR mice, as well as C57BL/6 mice treated with low doses of clodronate liposomes (ClLip), with Plasmodium chabaudi AS (Pc) parasites. The first evidence suggesting that DCs could contribute directly to parasite clearance was an early effect of the DTx treatment, but not of the ClLip treatment, in parasitemia control. DCs were also required for CD4+ T cell responses during infection. The phagocytosis of infected red blood cells (iRBCs) by splenic DCs was analyzed by confocal intravital microscopy, as well as by flow cytometry and immunofluorescence, at three distinct phases of Pc malaria: at the first encounter, at pre-crisis concomitant with parasitemia growth and at crisis when the parasitemia decline coincides with spleen closure. In vivo and ex vivo imaging of the spleen revealed that DCs actively phagocytize iRBCs and interact with CD4+ T cells both in T cell-rich areas and in the RP. Subcapsular RP DCs were highly efficient in the recognition and capture of iRBCs during pre-crisis, while complete DC maturation was only achieved during crisis. These findings indicate that, beyond their classical role in antigen presentation, DCs also contribute to the direct elimination of iRBCs during acute Plasmodium infection.São Paulo Research Foundation grants: (2011/24038-1 [MRDL], 2009/08559-1 [HBdS], CAPES/IGC 04/ 2012 [MRDL, CET])

Family network of children with special health needs: implications for Nursing

OBJECTIVE: to describe appropriate sources and resources for caregivers of children with special health needs in the community. METHOD: A qualitative study that used the creativity and sensitivity dynamics speaking map, part of the sensitive creative method, involving 11 caregivers of children with special health needs who are assisted in a university hospital located in the South of Brazil. RESULTS: the maps graphically represented through the genogram and ecomap showed that the caregiving women consistently and regularly use the resources of the internal and external family network; they eventually and irregularly access the community social network for physical and psychological support. CONCLUSION: the reclusive nature of care for these children inside the family circle contributes to their social invisibility. Based on this new information, it is recommended that Nursing participate in the care that is focused on these children's families, with particular attention to their socio-cultural conditions