ABSTRACT The 18S rRNA gene ( Rns ) phylogeny of Acanthamoeba is being investigated as a basis for improvements in the nomenclature and taxonomy genus. We previously analyzed sequences from 18 isolates morphological groups 2 3 found that they fell into four distinct evolutionary lineages we called sequence types T1‐T4. Here, 53 representing 16 species including 35 new strains. Eight additional (sequence T5‐T12) were identified. Four 12 included strains more than one nominal species. Thus,...

ABSTRACT Classification of Acanthamoeba at the subgenus level has been problematic, but increasing reports as an opportunistic human pathogen have generated interest in finding a more consistent basis for classification. Thus, we are developing classification scheme based on RNA gene sequences. This first report is analysis complete sequences nuclear small ribosomal subunit genes ( Rns ) from 18 strains. Sequence variation was localized 12 highly variable regions. Four distinct sequence...

ABSTRACT Acanthamoeba keratitis (AK) is a rare but sight-threatening ocular infection. Outbreaks have been associated with contaminated water and contact lens wear. The epidemiology pathology may be unique genotypes. We determined the Rns genotype for 37 clinical isolates from 23 patients presenting at University of Miami Bascom Palmer Eye Institute confirmed AK infections in 2006 to 2008. genus-specific ASA.S1 amplicon allowed rapid genotyping nonaxenic cultures. Of isolates, 36 were T4...

Abstract Matrix metalloproteinase‐9 (MMP‐9) is a well‐known regulator and effecter of many cellular processes including wound healing. In the cornea, either too much or little MMP‐9 can be detrimental to overall repair. We investigated secreted factors as well intracellular signaling pathways promoter sequences that mediate this regulation. Primary culture rabbit corneal epithelial cells were treated with various cytokines alone in different combinations induction was assessed by gel...

Uncontrolled increases of matrix metalloproteinase-9 (MMP-9) activity have been causally linked to epithelial barrier disruption and severe symptoms inflammatory diseases such as dry eye (DE). The data presented here show that the anti-inflammatory, cytoprotective intracellular extracellular chaperone protein clusterin (CLU) interacts with MMP-9 both inside outside cells. CLU bound very strongly active MMP-9, an affinity constant K(D) 2.63 nmol/L. Unexpectedly, had a much higher for...

Children with sepsis and multisystem organ failure have downregulated leukocyte gene expression of peroxisome proliferator-activated receptor-α (PPARα), a nuclear hormone receptor transcription factor that regulates inflammation lipid metabolism. Mouse models likewise demonstrated the absence PPARα is associated decreased survival injury, specifically heart. Using clinically relevant mouse model early sepsis, we found heart function increases in wild-type (WT) mice over first 24 h but...

Anesthetics used in infants and children are implicated the development of neurocognitive disorders. Although propofol induces neuroapoptosis developing brain, underlying mechanisms require elucidation may have an energetic basis. We studied substrate utilization immature swine anesthetized with either or isoflurane for 4 hours. Piglets were infused 13-Carbon-labeled glucose leucine common carotid artery to assess citric acid cycle (CAC) metabolism parietal cortex. The anesthetics produced...

Introduction Kawasaki disease (KD) is a diffuse vasculitis in children. Response to high dose intravenous gamma globulin (IVIG), the primary treatment, varies according genetic background. We sought identify loci, which associate with treatment response using whole genome sequencing (WGS). Method performed WGS 472 KD patients 305 IVIG responders and 167 non-responders defined by AHA clinical criteria. conducted logistic regression models test additive effect entire cohort four subgroups...

Aging presumably initiates shifts in substrate oxidation mediated part by changes insulin sensitivity. Similar occur with cardiac hypertrophy and may contribute to contractile dysfunction. We tested the hypothesis that aging modifies utilization alters sensitivity mouse heart when provided multiple substrates. In vivo function was measured microtipped pressure transducers left ventricle from control (4-6 mo) aged (22-24 mice. Cardiac also isolated working hearts along anaplerotic fractional...

Antifungal drugs have been proposed as a novel treatment for Acanthamoeba keratitis. The cysticidal activity of several antifungal compounds was tested against different genotypes culture collection and clinical isolates Acanthamoeba. Only voriconazole posaconazole were found to be cysticidal, with no differences in observed between isolates.

Background Protein posttranslational modifications by O ‐linked β‐N‐acetylglucosamine (O‐GlcNAc) increase with cardiac hypertrophy, yet the functional effects of these changes are incompletely understood. In other organs, O‐GlcNAc promotes adaptation to acute physiological stressors; however, prolonged elevations believed be detrimental. We hypothesize that early O‐GlcNAcylation improves function during initial response pressure overload but sustained established pathological hypertrophy...

purpose. This work was intended to test the classification of Acanthamoeba into genotypes based on nuclear ribosomal RNA gene (18S rDNA, Rns) sequences. Nearly all keratitis (AK) isolates are genotype RnsT4. marked phylogenetic localization is presumably either due an innate potential for pathogenicity or a peculiarity sequences used. To differentiate between these possibilities, relationships among have been reexamined, using second gene. methods. Phylogenetic were studied, mitochondrial...

Background Extracorporeal membrane oxygenation ( ECMO ) provides a bridge to recovery after myocardial injury in infants and children, yet morbidity mortality remain high. Weaning from the circuit requires adequate cardiac contractile function, which can be impaired by metabolic disturbances induced either ischemia‐reperfusion and/or . We tested hypothesis that although partially ameliorates abnormalities , these persist or recur with weaning. also determined if thyroid hormone...

Fungal ocular infections cause significant morbidity, particularly when diagnosis and treatment are delayed. Accurate morphologic identification of Fusarium spp. beyond the genus is time-consuming insensitive. It was purpose this study to examine usefulness nuclear ribosomal RNA (rRNA) internal transcribed spacer regions (ITS1 -2) detect differentiate responsible for infections.Fifty-eight archived isolates from sources 52 patients diagnosed with keratitis at Bascom Palmer Eye Institute...

Hypertrophic stimuli cause transcription of the proto-oncogene c-Myc (Myc). Prior work showed that myocardial knockout (Myc) attenuated hypertrophy and decreased expression metabolic genes after aortic constriction. Accordingly, we assessed interplay between Myc, substrate oxidation cardiac function during early pressure overload hypertrophy. Mice with specific, inducible Myc (MycKO-TAC) non-transgenic littermates (Cont-TAC) were subjected to transverse constriction (TAC; n = 7/group)....

Galactokinase is an essential enzyme in the metabolism of galactose. Patients with deficiencies galactokinase exhibit early-onset cataracts. We examined sequence human gene (GK1) from 13 patients exhibiting deficiency and identified 12 novel mutations. One mutations occurred six probands examined, remaining 11 were unique Expression each mutant GK1 genes Xenopus oocytes resulted very low activity levels. These results provide important information regarding types that occur population.

Cdk5 is a member of the cyclin-dependent kinase family, which expressed predominantly in terminally differentiated neurons. Lower levels are also found wide variety cell types, including lens. Although has been shown to play an important role neuronal migration and neurite outgrowth, its function non-neuronal cells not known. Therefore, this study was undertaken explore Results showed that, within adult mouse lens, localized cytoplasm,especially along lateral membranes differentiating...

Extracorporeal membrane oxygenation (ECMO) provides essential mechanical circulatory support necessary for survival in infants and children with acute cardiac decompensation. However, ECMO also causes metabolic disturbances, which contribute to total body wasting protein loss. Cardiac stunning can occur, prevents weaning, contributes high mortality. The heart may specifically undergo impairments, influence functional recovery. We tested the hypothesis that alters oxidative metabolism...

Protein posttranslational modifications (PTMs) by O-linked β-N-acetylglucosamine (O-GlcNAc) rise during pressure-overload hypertrophy (POH) to affect hypertrophic growth. The hexosamine biosynthesis pathway (HBP) branches from glycolysis make the moiety for O-GlcNAcylation. It is speculated that greater glucose utilization POH augments HBP flux increase O-GlcNAc levels; however, recent results suggest availability does not primarily regulate cardiac levels. We hypothesize induction of key...

Protein posttranslational modifications (PTMs) by O-GlcNAc globally rise during pressure-overload hypertrophy (POH). However, a major knowledge gap exists on the specific proteins undergoing changes in levels POH primarily because this PTM is low abundance and easily lost standard mass spectrometry (MS) conditions used for protein identification. Methodologies have emerged to enrich samples O-GlcNAcylated prior MS analysis. Accordingly, our goal was identify POH. We C57/Bl6 mice subjected...