A pipette-tip based method for seeding cells to droplet microfluidic platforms

Amongst various microfluidic platform designs frequently used for cellular analysis, droplet-microfluidics provides a robust tool for isolating and analyzing cells at the single-cell level by eliminating the influence of external factors on the cellular microenvironment. Encapsulation of cells in droplets is dictated by the Poisson distribution as a function of the number of cells present in each droplet and the average number of cells per volume of droplet. Primary cells, especially immune cells, or clinical specimens can be scarce and loss-less encapsulation of cells remains challenging. In this paper, we present a new methodology that uses pipette-tips to load cells to droplet-based microfluidic devices without the significant loss of cells. With various cell types , we demonstrate efficient cell encapsulation in droplets that closely corresponds to the encapsulation efficiency predicted by the Poisson distribution. Our method ensures loss-less loading of cells to microfluidic platforms and can be easily adapted for downstream single cell analysis, e.g., to decode cellular interactions between different cell types

Plasticity in nest site choice behavior in response to hydric conditions in a reptile

Natural selection is expected to select for and maintain maternal behaviors associated with choosing a nest site that promotes successful hatching of offspring, especially in animals that do not exhibit parental care such as reptiles. In contrast to temperature effects, we know little about how soil moisture contributes to successful hatching and particularly how it shapes nest site choice behavior in nature. The recent revelation of exceptionally deep nesting in lizards under extreme dry conditions underscored the potential for the hydric environment in shaping the evolution of nest site choice. But if deep nesting is an adaptation to dry conditions, is there a plastic component such that mothers would excavate deeper nests in drier years? We tested this hypothesis by excavating communal warrens of a large, deep-nesting monitor lizard (Varanus panoptes), taking advantage of four wet seasons with contrasting rainfall amounts. We found 75 nests during two excavations, including 45 nests after a 4-year period with larger wet season rainfall and 30 nests after a 4-year period with smaller wet season rainfall. Mothers nested significantly deeper in years associated with drier nesting seasons, a finding best explained as a plastic response to soil moisture, because differences in both the mean and variance in soil temperatures between 1 and 4 m deep are negligible. Our data are novel for reptiles in demonstrating plasticity in maternal behavior in response to hydric conditions during the time of nesting. The absence of evidence for other ground-nesting reptile mothers adjusting nest depth in response to a hydric-depth gradient is likely due to the tradeoff between moisture and temperature with changing depth; most ground-nesting reptile eggs are deposited at depths of ~ 2–25 cm—nesting deeper within or outside of that range of depths to achieve higher soil moisture would also generally create cooler conditions for embryos that need adequate heat for successful development. In contrast, extreme deep nesting in V. panoptes allowed us to disentangle temperature and moisture. Broadly, our data suggest that ground-nesting reptiles can assess soil moisture and respond by adjusting the depth of the nest, but may not, due to the cooling effect of nesting deeper. Our results, within the context of previous work, provide a more complete picture of how mothers can promote hatching success through adjustments in nest site choice behavior

Global Research Priorities to Better Understand the Burden of Iatrogenic Harm in Primary Care: An International Delphi Exercise

There is a need to identify and reach agreement on key foci for patient safety research in primary care contexts and understand how these priorities differ between low-, middle-, and high-income settings. We conducted a modified Delphi exercise, which was distributed to an international panel of experts in patient safety and primary care. Family practice and pharmacy were considered the main contexts on which to focus attention in order to advance patient safety in primary care across all income categories. Other clinical contexts prioritised included community midwifery and nursing in low-income countries and care homes in high-income countries. The sources of patient safety incidents requiring further study across all economic settings that were identified were communication between health care professionals and with patients, teamwork within the health care team, laboratory and diagnostic imaging investigations, issues relating to data management, transitions between different care settings, and chart/patient record com- pleteness. This work lays the foundation for a range of research initiatives that aim to promote a more comprehensive appreciation of the burden of unsafe primary care, develop understanding of the main areas of risk, and identify interventions that can enhance the safety of primary care provision internationall

UVA/UVA1 phototherapy and PUVA photochemotherapy in connective tissue diseases and related disorders: a research based review

BACKGROUND: Broad-band UVA, long-wave UVA1 and PUVA treatment have been described as an alternative/adjunct therapeutic option in a number of inflammatory and malignant skin diseases. Nevertheless, controlled studies investigating the efficacy of UVA irradiation in connective tissue diseases and related disorders are rare. METHODS: Searching the PubMed database the current article systematically reviews established and innovative therapeutic approaches of broad-band UVA irradiation, UVA1 phototherapy and PUVA photochemotherapy in a variety of different connective tissue disorders. RESULTS: Potential pathways include immunomodulation of inflammation, induction of collagenases and initiation of apoptosis. Even though holding the risk of carcinogenesis, photoaging or UV-induced exacerbation, UVA phototherapy seems to exhibit a tolerable risk/benefit ratio at least in systemic sclerosis, localized scleroderma, extragenital lichen sclerosus et atrophicus, sclerodermoid graft-versus-host disease, lupus erythematosus and a number of sclerotic rarities. CONCLUSIONS: Based on the data retrieved from the literature, therapeutic UVA exposure seems to be effective in connective tissue diseases and related disorders. However, more controlled investigations are needed in order to establish a clear-cut catalogue of indications

Ecosystem engineering by deep-nesting monitor lizards

As the current biodiversity crisis approaches levels comparable to the rates of the five historical mass extinctions, increasing attention has focused on how to stop or slow species loss and preserve ecosystem function. The impact of the loss of an individual species on communities and ecosystems is heterogeneous, however. Removing some species has negligible effects while the removal of others can be catastrophic. Metaphorically, the scenario can be likened to Jenga, a popular block-balancing game in which players build a tower of wooden pieces, analogous to a dynamic ecosystem (de Ruiter et al. 2005)