Inducible ablation of CD11c+ cells to determine their role in skin wound repair

Wound healing in healthy skin requires a complex interplay between immune and non‐immune cells. In addition to their roles in infection control and cell debridement, leukocytes secrete factors to orchestrate the timing of the repair process via cross‐talk with epithelia. This critical role in wound timing is vital as failure to induce wound closure leads to debilitating chronic wounds susceptible to further infection and patient sepsis

The antibacterial activity of a photoactivatable diarylacetylene against Gram-positive bacteria

The emergence of antibiotic resistance is a growing threat to human health, and therefore, alternatives to existing compounds are urgently needed. In this context, a novel fluorescent photoactivatable diarylacetylene has been identified and characterised for its antibacterial activity, which preferentially eliminates Gram-positive over Gram-negative bacteria. Experiments confirmed that the Gram-negative lipopolysaccharide-rich outer surface is responsible for tolerance, as strains with reduced outer membrane integrity showed increased susceptibility. Additionally, bacteria deficient in oxidative damage repair pathways also displayed enhanced sensitivity, confirming that reactive oxygen species production is the mechanism of antibacterial activity. This new diarylacetylene shows promise as an antibacterial agent against Gram-positive bacteria that can be activated in situ, potentially for the treatment of skin infections

The antibacterial activity of a photoactivatable diarylacetylene against Gram-positive bacteria

The emergence of antibiotic resistance is a growing threat to human health, and therefore, alternatives to existing compounds are urgently needed. In this context, a novel fluorescent photoactivatable diarylacetylene has been identified and characterised for its antibacterial activity, which preferentially eliminates Gram-positive over Gram-negative bacteria. Experiments confirmed that the Gram-negative lipopolysaccharide-rich outer surface is responsible for tolerance, as strains with reduced outer membrane integrity showed increased susceptibility. Additionally, bacteria deficient in oxidative damage repair pathways also displayed enhanced sensitivity, confirming that reactive oxygen species production is the mechanism of antibacterial activity. This new diarylacetylene shows promise as an antibacterial agent against Gram-positive bacteria that can be activated in situ, potentially for the treatment of skin infections

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Heat-denaturation removes key serum factors needed to support epidermal stratification.

<p>(<b>A–C</b>) Eight micron sections of p4 NHEKs grown on DEDs in complete FAD (A), KGM-Gold (B) or EpiLife (C) supplemented with 10% serum boiled at 95°C and 1.4 mM Ca⁺⁺. Scale bars: 50 microns.</p

Keratinocyte media sufficient to support feeder-free, serum-free epidermal growth fails to support stratification in skin equivalents.

<p>(<b>A, B, D–F</b>) Hematoxylin and eosin-stained sections of p4 NHEKs grown on DEDs in the following culture media: (A) complete FAD (high Ca⁺⁺/10%FCS), (B) KGM-Gold (low Ca⁺⁺), (D) low-Ca⁺⁺ FAD with 10% serum (low Ca⁺⁺/10%FCS), (E) KGM-gold plus added Ca⁺⁺ (high Ca⁺⁺), (F) KGM-Gold supplemented with 10% fetal bovine serum (low Ca⁺⁺/10%FCS) and (G) KGM supplemented with 10% fetal bovine serum and added Ca⁺⁺ (high Ca⁺⁺/10%FCS). Insets show higher magnification images. Layers of the stratified epidermis indicated: B, basal; S, spinous; G, granular; C, cornified. (<b>C</b>) Graph indicates the average depth of the epithelium layer in the different types of media. Statistical significance calculated relative to control, complete FAD (high Ca⁺⁺/10%FCS) cultured samples. Black arrow in B marks keratinocytes on DED surface. Blue arrows in A and G marks granular cells. ** p value<0.005; *** p value<0.0005 (Student's t-test); scale bars: 50 microns.</p

Heat-treating serum increases epidermal thickness and organisation.

<p>Eight micron sections of p4 NHEKs grown on DEDs in complete FAD with 10% serum heated to 56°C (A, D, F; FAD+HI serum) or KGM supplemented with 10% serum heated to 56°C and added Ca⁺⁺ (B, E, G; KGM+HI serum). (<b>A, B, D–H</b>) Sections were stained with hematoxylin and eosin or with antibodies to Keratin 14 (D, E, green) or Keratin 10 (F, G, green). Sections were counterstained with DAPI (blue). (<b>C</b>) Graph indicates the average depth of the epithelium layer in the two types of media. Statistical significance calculated relative to control, complete FAD with 10% serum (high Ca⁺⁺/10%FCS) cultured samples (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0052494#pone-0052494-g002" target="_blank">Figure 2</a>). ** p value<0.005; *** p value<0.0005 (Student's t-test); scale bars: 50 microns.</p

EpiLife media supplemented with calcium and 10% serum supports keratinocyte stratification.

<p>(<b>A–C</b>) Hematoxylin and eosin-stained sections of p4 NHEKs grown on DEDs in the following culture media: (A) EpiLife plus added Ca⁺⁺ (high Ca⁺⁺), (B) EpiLife supplemented with 10% fetal bovine serum (low Ca⁺⁺/10%FCS) or (C) EpiLife supplemented with 10% fetal bovine serum and added Ca⁺⁺ (high Ca⁺⁺/10%FCS). Scale bars: 50 microns.</p