Are Thermotolerant and Osmotolerant Characteristics of Acanthamoeba Species an Indicator of Pathogenicity?

Objective: The aim of this study was to evaluate the pathogenicity of Acanthamoeba strains with T4, T5, T11, and T12 genotypes by comparing the osmotolerance and thermotolerance characteristics of Acanthamoeba strains isolated from genotype groups, within species with the same genotype, and from environmental and keratitis cases. Methods: In this study, after axenic cultures of 22 Acanthamoeba strains with T4 (Neff, A, B, D, E), T5, T11, and T12 genotypes isolated from clinical and environmental samples, thermotolerance (37 °C, 39 °C and 41 °C) and osmotolerance (0.5 M, 1 M) tests were performed. Results: All strains showed growth ability at 37 °C and 0.5 M osmolarity. While all five strains isolated from patients with Acanthamoeba keratitis showed growth ability at 37 °C and 0.5 M osmolarity, no growth was detected at 41 °C and 1 M osmolarity. When the tolerance characteristics of the strains with the same genotype were evaluated, the strains with the T5 and T4E genotypes showed the same characteristics. When Acanthamoeba strains with the T4 genotype were evaluated in general, 31.25% of the strains were found to grow at 39 °C and 6.25% at 41 °C. Of the T4Neff strains, only one strain did not show the ability to reproduce at 39 °C and showed a different feature from the other strains. While the strain with the T11 genotype grew at all temperatures, the strain with the T12 genotype did not grow at 41 °C. Conclusion: According to our research results, we believe that tolerance to 39 °C and 1 M mannitol is not an indicator of pathogenicity. More studies with Acanthamoeba strains are required to clarify this issue.

Single-cell and bulk transcriptome sequencing identifies two epithelial tumor cell states and refines the consensus molecular classification of colorectal cancer.

The consensus molecular subtype (CMS) classification of colorectal cancer is based on bulk transcriptomics. The underlying epithelial cell diversity remains unclear. We analyzed 373,058 single-cell transcriptomes from 63 patients, focusing on 49,155 epithelial cells. We identified a pervasive genetic and transcriptomic dichotomy of malignant cells, based on distinct gene expression, DNA copy number and gene regulatory network. We recapitulated these subtypes in bulk transcriptomes from 3,614 patients. The two intrinsic subtypes, iCMS2 and iCMS3, refine CMS. iCMS3 comprises microsatellite unstable (MSI-H) cancers and one-third of microsatellite-stable (MSS) tumors. iCMS3 MSS cancers are transcriptomically more similar to MSI-H cancers than to other MSS cancers. CMS4 cancers had either iCMS2 or iCMS3 epithelium; the latter had the worst prognosis. We defined the intrinsic epithelial axis of colorectal cancer and propose a refined 'IMF' classification with five subtypes, combining intrinsic epithelial subtype (I), microsatellite instability status (M) and fibrosis (F).

The Effect of Dream Anxiety on Sleep Quality in Inflammatory Bowel Diseases.

INTRODUCTION: Declining sleep quality is a well-known issue in inflammatory bowel disease (IBD), but dream characteristics of patients with IBD and their role in sleep quality are unknown. In this study, we aimed to examine whether and how patients with ulcerative colitis (UC) and Crohn's disease (CD) differ on sleep quality, sleepiness level, and dream anxiety (DA) level compared to healthy controls (HC), controlling for their depressive and anxious tendencies. METHODS: Patients and HCs were enrolled prospectively into the study. The Van DA Scale, Pittsburg Sleep Quality Index, Epworth Sleepiness Scale, Beck Depression Index, and State-Trait Anxiety Inventories were used to assess DA, sleep quality, sleepiness, depression, and anxiety, respectively. RESULTS: Patients with IBD had significantly lower depression (p = 0.004), state anxiety (p = 0.0001), trait anxiety (p = 0.004), and DA (p = 0.0001) than HCs. Although no statistically significant difference in sleep quality was found (p = 0.99), daytime sleepiness was more common in HCs than in IBD patients (p = 0.0001). No statistically significant difference was seen in depression, state anxiety, trait anxiety, DA, sleep quality, and daytime sleepiness between patients with CD and those with UC. No correlation was found between disease activity indices and psychological parameters. CONCLUSION: In contrast to previous studies, this study found lower anxiety and depression levels in patients with IBD than in HCs. Moreover, DA score was higher in HCs. For the first time, we revealed that DA may be one of the factors leading to sleep disturbance in patients with IBD.

The effects of antibiotics and melatonin on hepato-intestinal inflammation and gut microbial dysbiosis induced by a short-term high-fat diet consumption in rats.

High-fat diet (HFD) consumption leads to metabolic disorders, gastrointestinal dysfunction and intestinal dysbiosis. Antibiotics also disrupt the composition of intestinal microbiota. The aim of the present study was to investigate the impact of a short-term feeding with HFD on oxidative status, enteric microbiota, intestinal motility and the effects of antibiotics and/or melatonin treatments on diet-induced hepato-intestinal dysfunction and inflammation. Male Sprague-Dawley rats were pair-fed with either standard chow or HFD (45 % fat) and were given tap water or melatonin (4 mg/kg per d) or melatonin plus antibiotics (ABX; neomycin, ampicillin, metronidazole; each 1 g/l) in drinking water for 2 weeks. On the 14th day, colonic motility was measured and the next day intestinal transit was assessed using charcoal propagation. Trunk blood, liver and intestine samples were removed for biochemical and histopathological evaluations, and faeces were collected for microbiota analysis. A 2-week HFD feeding increased blood glucose level and perirenal fat weight, induced low-level hepatic and intestinal inflammation, delayed intestinal transit, led to deterioration of epithelial tight junctions and overgrowth of colonic bacteria. Melatonin intake in HFD-fed rats reduced ileal inflammation, colonic motility and perirenal fat accumulation. ABX abolished increases in fat accumulation and blood glucose, reduced ileal oxidative damage, suppressed HFD-induced overgrowth in colonic bacteria, and reversed HFD-induced delay in intestinal transit; however, hepatic neutrophil accumulation, hepatic injury and dysfunction were further enhanced. In conclusion, the results demonstrate that even a short-term HFD ingestion results in hepato-intestinal inflammatory state and alterations in bacterial populations, which may be worsened with antibiotic intake, but alleviated by melatonin.

The polymorphisms of the MBL2 and MIF genes associated with Pediatric Cochlear Implant Patients.

OBJECTIVES: Mannose-binding lectin and macrophage migration inhibitory factor gene polymorphisms are associated with several acute/chronic autoimmune or inflammatory diseases. The aim of this study was to investigate if there was any association between mannose-binding lectin 2 (MBL2) and macrophage migration inhibitory factor (MIF) gene polymorphisms and profound congenital sensorineural hearing loss in children who underwent cochlear implantation. METHODS: A total of 62 patients with congenital hearing loss and 80 age- and sex-matched healthy controls were evaluated for codon 54 A/B polymorphisms in MBL2 and the-173 G/C polymorphism in MIF by using the polymerase chain reaction and restriction fragment length polymorphism method. RESULTS: The frequency of the BB genotype of MBL2 and MIF -173 GC genotype were statistically significantly higher in the patient group than in the controls (p=0.0127, p=0.0408, respectively). CONCLUSION: In this study, we found that a subject who is homozygous for the variant allele B of codon 54 of the MBL2and heterozygous for variant allele C of -173 MIF has a risk factor for sensorineural hearing loss.