Parakeratosis

A mature dairy cow was transitioned from a high forage (100% forage) to a high-grain (79% grain) diet over seven days. Continuous ruminal pH recordings were utilized to diagnose the severity of ruminal acidosis. Additionally, blood and rumen papillae biopsies were collected to describe the structural and functional adaptations of the rumen epithelium. On the final day of the grain challenge, the daily mean ruminal pH was 5.41+/-0.09 with a minimum of 4.89 and a maximum of 6.31. Ruminal pH was under 5.0 for 130 minutes (2.17 hours) which is characterized as the acute form of ruminal acidosis in cattle. The grain challenge increased blood beta-hydroxybutyrate by 1.8 times and rumen papillae mRNA expression of 3-hydroxy-3-methylglutaryl-coenzyme A synthase by 1.6 times. Ultrastructural and histological adaptations of the rumen epithelium were imaged by scanning electron and light microscopy. Rumen papillae from the high grain diet displayed extensive sloughing of the stratum corneum and compromised cell adhesion as large gaps were apparent between cells throughout the strata. This case report represents a rare documentation of how the rumen epithelium alters its function and structure during the initial stage of acute acidosis.

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 Δep ), Dnase1l2 -/- Dnase2 Δep 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 Δep 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 Δep 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.

Hereditary nasal parakeratosis (HNPK), an inherited monogenic autosomal recessive skin disorder, leads to crusts and fissures on the nasal planum of Labrador Retrievers. We performed a genome-wide association study (GWAS) using 13 HNPK cases and 23 controls. We obtained a single strong association signal on chromosome 2 (p(raw) = 4.4×10⁻¹⁴). The analysis of shared haplotypes among the 13 cases defined a critical interval of 1.6 Mb with 25 predicted genes. We re-sequenced the genome of one case at 38× coverage and detected 3 non-synonymous variants in the critical interval with respect to the reference genome assembly. We genotyped these variants in larger cohorts of dogs and only one was perfectly associated with the HNPK phenotype in a cohort of more than 500 dogs. This candidate causative variant is a missense variant in the SUV39H2 gene encoding a histone 3 lysine 9 (H3K9) methyltransferase, which mediates chromatin silencing. The variant c.972T>G is predicted to change an evolutionary conserved asparagine into a lysine in the catalytically active domain of the enzyme (p.N324K). We further studied the histopathological alterations in the epidermis in vivo. Our data suggest that the HNPK phenotype is not caused by hyperproliferation, but rather delayed terminal differentiation of keratinocytes. Thus, our data provide evidence that SUV39H2 is involved in the epigenetic regulation of keratinocyte differentiation ensuring proper stratification and tight sealing of the mammalian epidermis.

Granular parakeratosis (GP) is an uncommon keratinization disorder that predominantly affects intertriginous areas and may clinically mimic atopic dermatitis or psoriasis. Histologically, GP is characterized by compact hyperkeratosis, parakeratosis, and retention of keratohyalin granules within the stratum corneum, although its histopathological spectrum is heterogeneous. Previous studies have consisted mainly of case reports, while mechanistic investigations remain limited, and whether GP represents a purely cornification-related abnormality is unclear. During and after the coronavirus disease 2019 (COVID-19) pandemic, increased use of benzalkonium chloride (BAC)-containing laundry disinfectants coincided with more patients presenting with recurrent erythema and scaling that responded poorly to conventional treatment, prompting renewed attention to this disease. We performed single-cell RNA sequencing of lesional skin from three patients with GP and matched skin samples from three healthy controls, followed by keratinocyte subclustering, pseudotime analysis, and cell-cell communication analysis. Key findings were further validated by immunofluorescence in an independent cohort of four patients with GP and three healthy controls. HaCaT cells were exposed to BAC and analyzed by the Cell Counting Kit-8 (CCK-8) assay and bulk RNA sequencing. Single-cell analysis revealed the expansion of inflammatory keratinocytes in GP lesions. Pathway enrichment showed the activation of innate immune and antimicrobial responses and dysregulation of keratinocyte differentiation. S100A8/S100A9 were increased in GP keratinocytes and validated at the protein level. BAC induced interleukin-1 (IL-1)-related inflammatory activation and cellular stress responses, suggesting that GP is an exposure-associated inflammatory epidermal differentiation disorder.