DNase1L2 Degrades Nuclear DNA during Corneocyte Formation

The removal of keratinocyte (KC) nuclear DNA by deoxyribonucleases (DNases) is an important step in the formation of normal stratum corneum (SC). However, the molecular identity of the DNA-degrading enzymes has so far remained elusive. Here we show that the endonuclease DNase1-like 2 (DNase1L2) is preferentially expressed in the epidermis and that its expression correlates with terminal differentiation of KC in vitro and in vivo. In biopsies of normal skin, DNase1L2 mRNA was regularly found in suprabasal KC and DNase1L2 protein was highly abundant in the stratum granulosum. In contrast to normal skin, DNase1L2 expression was downregulated in parakeratotic epidermis such as in psoriatic lesions. When DNase1L2 gene expression was knocked down by small interfering RNA in a human skin equivalent model, nuclei were maintained through all layers of the SC. Taken together, our data demonstrate that DNase1L2 plays an essential role in DNA degradation during terminal differentiation of epidermal KC

Community shifts from eukaryote to cyanobacteria dominated phytoplankton: The role of mixing depth and light quality

Lake stratification strengthens with increasing surface water temperatures, thereby reducing the depth of the mixed layer. Phytoplankton communities are not only exposed to different nutrient availability within a mixed water column, but also to different light quality. We conducted controled laboratory and mesocosm experiments to investigate phytoplankton, especially cyanobacteria, responses to different light quality and mixing depths. Our mesocosm experiment allowed the manipulation of mixing depth in situ by a mesocosm approach and to follow the effects of changing mixing depth on the phytoplankton community composition. Our laboratory experiment allowed the control of temperature and light quantity. To investigate the effect of light quality on phytoplankton, we created a light gradient from full photosynthetic active radiation to a reduced blue spectrum. In both experiments, shifts in phytoplankton community composition from eukaryote to cyanobacteria occurred at shallow mixing depth with higher availability of photosynthetic active radiation. Our results from the mesocosm experiment support the idea that reduced mixing depth can promote cyanobacterial abundance. With our laboratory experiment, we were able to manipulate light quality independent of temperature, available nutrients and light intensity influencing phytoplankton abundance. Results from the laboratory experiments support our hypothesis that a shift in light spectrum alone is a driver, strong enough to enhance cyanobacteria occurrence. Most of the previous studies dealing with cyanobacterial blooms have investigated temperature and eutrophication effects. Certainly, these are major factors for the growth of phytoplankton, but our experiments show that other aspects, such as the quality of light, must be also taken into account to explain cyanobacterial blooms. Such shifts in the phytoplankton community from eukaryote to cyanobacteria dominated communities will have strong consequences for food web dynamics. Several cyanobacteria specific traits, (e.g., toxin production, lack of essential fatty acids, and inedibility through production of large colonies) reduce transfer efficiencies of energy and matter between phyto- and zooplankton and therefore can influence higher trophic levels such as fish

Fast Humidity Sensors for Harsh Environment

With the application of a recently developed deposition method called initiated chemical vapor deposition (iCVD), remarkably fast responsive hydrogel thin films in the order of a few hundred nanometers were created. When in contact with humid air, the hydrogel layer extends its thickness manifold, which can be detected. The verification of the thickness change was realized interferometrically with a laser and a white light input source in two different implementations. The setup was designed without electric components in the vicinity of the active sensor layer and is therefore applicable in harsh and explosive environment. The achieved response time for an abrupt change of the humidity τ63 ≤ 2.5 s is about three times lower compared to one of the fastest commercially available sensors on the market

Fast Optical Humidity Sensor Based on Hydrogel Thin Film Expansion for Harsh Environment

With the application of a recently developed deposition method called initiated chemical vapor deposition (iCVD), responsive hydrogel thin films in the order of a few hundred nanometers were created. When in contact with humid air, the hydrogel layer increases its thickness considerably. The measurement of the thickness change was realized interferometrically with a laser and a broadband light source in two different implementations. The relative change in thickness with respect to humidity can be described with the Flory⁻Huggins theory. The required Flory⁻Huggins interaction parameter was determined for the actual hydrogel composition. The setup was designed without electric components in the vicinity of the active sensor layer and is therefore applicable in harsh environments such as explosive or corrosive ones. The implemented sensor prototype delivered reproducible relative humidity ( R H ) values and the achieved response time for an abrupt change of the humidity τ 63 ≤ 2.5 s was about three times faster compared to one of the fastest commercially available sensors on the market

The Trichohyalin-Like Protein Scaffoldin Is Expressed in the Multilayered Periderm during Development of Avian Beak and Egg Tooth

Scaffoldin, an S100 fused-type protein (SFTP) with high amino acid sequence similarity to the mammalian hair follicle protein trichohyalin, has been identified in reptiles and birds, but its functions are not yet fully understood. Here, we investigated the expression pattern of scaffoldin and cornulin, a related SFTP, in the developing beaks of birds. We determined the mRNA levels of both SFTPs by reverse transcription polymerase chain reaction (RT-PCR) in the beak and other ectodermal tissues of chicken (Gallus gallus) and quail (Coturnix japonica) embryos. Immunohistochemical staining was performed to localize scaffoldin in tissues. Scaffoldin and cornulin were expressed in the beak and, at lower levels, in other embryonic tissues of both chickens and quails. Immunohistochemistry revealed scaffoldin in the peridermal compartment of the egg tooth, a transitory cornified protuberance (caruncle) on the upper beak which breaks the eggshell during hatching. Furthermore, scaffoldin marked a multilayered peridermal structure on the lower beak. The results of this study suggest that scaffoldin plays an evolutionarily conserved role in the development of the avian beak with a particular function in the morphogenesis of the egg tooth

Scientific Reports / Inactivation of DNase1L2 and DNase2 in keratinocytes suppresses DNA degradation during epidermal cornification and results in constitutive parakeratosis

The stratum corneum of the epidermis constitutes the mammalian skin barrier to the environment. It is formed by cornification of keratinocytes, a process which involves the removal of nuclear DNA. Here, we investigated the mechanism of cornification-associated DNA degradation by generating mouse models deficient of candidate DNA-degrading enzymes and characterizing their epidermal phenotypes. In contrast to Dnase1l2 / mice and keratinocyte-specific DNase2 knockout mice (Dnase2 ep), Dnase1l2 / Dnase2 ep mice aberrantly retained nuclear DNA in the stratum corneum, a phenomenon commonly referred to as parakeratosis. The DNA within DNase1L2/DNase2-deficient corneocytes was partially degraded in a DNase1-independent manner. Isolation of corneocytes, i.e. the cornified cell components of the stratum corneum, and labelling of DNA demonstrated that corneocytes of Dnase1l2 / Dnase2 ep mice contained DNA in a nucleus-shaped compartment that also contained nucleosomal histones but lacked the nuclear intermediate filament protein lamin A/C. Parakeratosis was not associated with altered corneocyte resistance to mechanical stress, changes in transepidermal water loss, or inflammatory infiltrates in Dnase1l2 / Dnase2 ep mice. The results of this study suggest that cornification of epidermal keratinocytes depends on the cooperation of DNase1L2 and DNase2 and indicate that parakeratosis per se does not suffice to cause skin pathologies.(VLID)461539

The Expression of the Endogenous mTORC1 Inhibitor Sestrin 2 Is Induced by UVB and Balanced with the Expression Level of Sestrin 1.

Sestrin 2 (SESN2) is an evolutionarily conserved regulator of mechanistic target of rapamycin complex 1 (mTORC1) which controls central cellular processes such as protein translation and autophagy. Previous studies have suggested that SESN2 itself is subjected to regulation at multiple levels. Here, we investigated the expression of SESN2 in the skin and in isolated skin cells. SESN2 was detected by immunofluorescence analysis in fibroblasts and keratinocytes of human skin. Differentiation of epidermal keratinocytes was not associated with altered SESN2 expression and siRNA-mediated knockdown of SESN2 did not impair stratum corneum formation in vitro. However, SESN2 was increased in both cell types when the expression of its paralog SESN1 was blocked by siRNA-mediated knock down, indicating a compensatory mechanism for the control of expression. Irradiation with UVB but not with UVA significantly increased SESN2 expression in both keratinocytes and fibroblasts. Upregulation of SESN2 expression could be completely blocked by suppression of p53. These results suggest that SESN2 is dispensable for normal epidermal keratinization but involved in the UVB stress response of skin cells

Inactivation of Autophagy in Keratinocytes Reduces Tumor Growth in Mouse Models of Epithelial Skin Cancer

Autophagy is a ubiquitous degradation mechanism, which plays a critical role in cellular homeostasis. To test whether autophagy suppresses or supports the growth of tumors in the epidermis of the skin, we inactivated the essential autophagy gene Atg7 specifically in the epidermal keratinocytes of mice (Atg7∆ep) and subjected such mutant mice and fully autophagy-competent mice to tumorigenesis. The lack of epithelial Atg7 did not prevent tumor formation in response to 7, 12-dimethylbenz(a)anthracene (DMBA) as the initiator and 12-O tetradecanoylphorbol-13-acetate (TPA) as the promoter of tumor growth. However, the number of tumors per mouse was reduced in mice with epithelial Atg7 deficiency. In the K5-SOS EGFRwa2/wa2 mouse model, epithelial tumors were initiated by Son of sevenless (SOS) in response to wounding. Within 12 weeks after tumor initiation, 60% of the autophagy-competent K5-SOS EGFRwa2/wa2 mice had tumors of 1 cm diameter and had to be sacrificed, whereas none of the Atg7∆ep K5-SOS EGFRwa2/wa2 mice formed tumors of this size. In summary, the deletion of Atg7 reduced the growth of epithelial tumors in these two mouse models of skin cancer. Thus, our data show that the inhibition of autophagy limits the growth of epithelial skin tumors