Fungal Keratitis in a Critically Ill Post-trauma Patient.

Keratitis is the leading cause of corneal blindness in the world. Nearly half the cases are due to a fungal infection known as fungal keratitis (FK). There is much variability in the clinical presentation of FK, so diagnosis can be difficult. With the risks of blindness in disease progression being so high, it is vital to diagnose and treat FK quickly. We present a case of FK due to Candida albicans and Staphylococcus lugdunensis-oxa ss after a motor vehicle accident, its treatment, and the general outcome. A 71-year-old man with a history of hypertension, hyperlipidemia, arthritis, and previous tobacco use presented after a helmeted motorcycle accident with back pain and bilateral lower extremity sensory and motor function loss. He suffered many fractures and was in neurogenic shock. He had nearly daily reduction and fixation of multiple axial spinal fractures while in the surgical intensive care unit and was ultimately unable to be successfully extubated. Between two intubations, he complained to his family of blurry vision, and there was notable purulence and corneal haziness in bilateral eyes. The healthcare team initially suspected the eye infection was due to a bacterial etiology, and he was subsequently diagnosed with Pseudomonas pneumonia on respiratory cultures. However, several days of antibiotics did not improve the ocular exam. A corneal culture was positive for C. albicans and S. lugdunensis-oxa ss, and anti-fungal treatment was initiated with ocular improvement. Unfortunately, the patient succumbed to his injuries and further sepsis at another site. With a progressively poor prognosis and machine dependence, he was made do-not-resuscitate per family wishes and died within two hours after cessation of hemodialysis. One of the greatest barriers to diagnosing FK in the United States is the absence of information regarding the disease. Though novel diagnoses and treatment strategies are in development, the fungal etiology of keratitis should be included in the curricula for not just medical students but also for providers and specialists, as the incidence of FK continues to grow with globalization. We also aim to emphasize the importance of a multidisciplinary team in these cases, as ophthalmology and infectious disease specialists should be involved immediately in order to improve patient outcomes.

Using poly(l-lactic acid) and poly(ɛ-caprolactone) blends to fabricate self-expanding, watertight and biodegradable surgical patches for potential fetoscopic myelomeningocele repair.

Our study focuses on the development and characterization of a self-expanding, watertight and biodegradable patch for fetoscopic myelomeningocele (MMC) prenatal repair. We fabricated poly(l-lactic acid) (PLA) and poly(ɛ-caprolactone) (PCL) blend films by solution casting. Formulation c with average glass transition temperature of 37.6 ± 1.2°C was chosen for temporospatial recovery. Favorable results from surface studies reflected homogeneous dispersion of polymers in the blend. The cytotoxicity was studied in human foreskin fibroblasts. The blend film was cytocompatible, evidenced by matching percentage of live cells in exposed and control solutions. Subsequently, liquid water permeability experiments confirmed watertight nature of films. Finally, in vitro degradation was investigated in phosphate buffered saline (PBS) and amniotic fluid (AF) separately for 16 weeks. Similar weight loss (n = 6, p = 0.912) and significantly different (n = 3, p = 0.025) surface roughness was observed in PBS and AF, respectively, at 16 weeks. Functional group analysis displayed increasing carbonyl and hydroxyl bonds in PBS and AF, respectively, over time, indicating progression of hydrolytic degradation. Favorable characterization results provide strong evidence to employ PLA-PCL blend films as surgical patches in fetoscopic MMC repair. Designed patch serves as standalone system to successfully tackle impending hurdles of MMC repair and proves to be a superior alternative compared to existing patches. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 295-305, 2019.

MAP kinases associate with high molecular weight multiprotein complexes.

Plant responses to the environment and developmental processes are mediated by a complex signaling network. The Arabidopsis thaliana mitogen-activated protein kinases (MAPKs) MPK3 and MPK6 and their orthologs in other plants are shared signal transducers that respond to many developmental and environmental signals and thus represent highly connected hubs in the cellular signaling network. In animals, specific MAPK signaling complexes are assembled which enable input-specific protein-protein interactions and thus specific signaling outcomes. In plants, not much is known about such signaling complexes. Here, we report that MPK3, MPK6, and MPK10 orthologs in tomato, tobacco, and Arabidopsis as well as tomato MAPK kinase 4 (MKK4) associate with high molecular weight (~250-550 kDa) multiprotein complexes. Elicitation by the defense-associated peptides flg22 and systemin resulted in phosphorylation and activation of the monomeric MAPKs, whereas the complex-associated MAPKs remained unphosphorylated and inactive. In contrast, treatment of tomato cells with a phosphatase inhibitor resulted in association of phosphorylated MPK1/2 with the complex. These results demonstrate that plant MAPKs and MAPKKs dynamically assemble into stable multiprotein complexes and this may depend on their phosphorylation status. Identification of the constituents of these multiprotein complexes promises a deeper understanding of signaling dynamics.

The COP9 signalosome controls jasmonic acid synthesis and plant responses to herbivory and pathogens.

The COP9 signalosome (CSN) is a multi-protein complex that regulates the activities of cullin-RING E3 ubiquitin ligases (CRLs). CRLs ubiquitinate proteins in order to target them for proteasomal degradation. The CSN is required for proper plant development. Here we show that the CSN also has a profound effect on plant defense responses. Silencing of genes for CSN subunits in tomato plants resulted in a mild morphological phenotype and reduced expression of wound-responsive genes in response to mechanical wounding, attack by Manduca sexta larvae, and Prosystemin over-expression. In contrast, expression of pathogenesis-related genes was increased in a stimulus-independent manner in these plants. The reduced wound response in CSN-silenced plants corresponded with reduced synthesis of jasmonic acid (JA), but levels of salicylic acid (SA) were unaltered. As a consequence, these plants exhibited reduced resistance against herbivorous M. sexta larvae and the necrotrophic fungal pathogen Botrytis cinerea. In contrast, susceptibility to tobacco mosaic virus (TMV) was not altered in CSN-silenced plants. These data demonstrate that the CSN orchestrates not only plant development but also JA-dependent plant defense responses.