The circulatory small non-coding RNA landscape in community-acquired pneumonia on intensive care unit admission

Community-acquired pneumonia (CAP) is a major cause of sepsis. Despite several clinical trials targeting components of the inflammatory response, no specific treatment other than antimicrobial therapy has been approved. This argued for a deeper understanding of sepsis immunopathology, in particular factors that can modulate the host response. Small non-coding RNA, for example, micro (mi)RNA, have been established as important modifiers of cellular phenotypes. Notably, miRNAs are not exclusive to the intracellular milieu but have also been detected extracellular in the circulation with functional consequences. Here, we sought to determine shifts in circulatory small RNA levels of critically ill patients with CAP-associated sepsis and to determine the influence of clinical severity and causal pathogens on small RNA levels. Blood plasma was collected from 13 critically ill patients with sepsis caused by CAP on intensive care unit admission and from 5 non-infectious control participants. Plasma small RNA-sequencing identified significantly altered levels of primarily mature miRNAs in CAP relative to controls. Pathways analysis of high or low abundance miRNA identified various over-represented cellular biological pathways. Analysis of small RNA levels against common clinical severity and inflammatory parameters indices showed direct and indirect correlations. Additionally, variance of plasma small RNA levels in CAP patients may be explained, at least in part, by differences in causal pathogens. Small nuclear RNA levels were specifically altered in CAP due to Influenza infection in contrast to Streptococcus pneumoniae infection. Pathway analysis of plasma miRNA signatures unique to Influenza or Streptococcus pneumoniae infections showed enrichment for specific proteoglycan, cell cycle, and immunometabolic pathways.     

Comparison of performance for leukocyte differential counting of the Technicon H6000 system with a manual reference method using the NCCLS standard

The National Committee for Clinical Laboratory Standards (NCCLS) has published a tentative standard for leukocyte differential counting, by means of which a manual or automated method for leukocyte differential counting can be compared with a manual reference method. The performance of the Technicon H6000 system was evaluated using the standard at Stamford and Overlook Hospitals. A total of 502 patient samples were analyzed: 315 from Overlook and 187 from Stamford. The H6000 system was found to be approximately four times more precise than the 200-cell manual reference method for each cell type. Correlation of the H6000 system with the manual method was good, with correlation coefficients of 0.98 for neutrophils and lymphocytes, 0.96 for eosinophils, 0.72 for monocytes, and 0.5 for basophils. The clinical sensitivity of the H6000 system, measured in terms of false normals and false abnormals, was similar to that of the manual reference method when measured against itself. There were no clinically significant discrepancies in results from the H6000 system, except for possibly one case where a patient was already on antibiotic therapy. The NCCLS standard was found to be a useful but rather complex and involved method for evaluating the performance of the H6000 system, the major problem being the amount of work needed to count manually the number of cells required for the manual reference method.     

Influence of prostaglandin F autoantibodies on the estrous cycle of the guinea pig

Adult female guinea pigs were actively immunized with prostaglandin F_2α conjugated to bovine serum albumin (BSA). Control animals, immunized against BSA continued to cycle normally, while the animals immunized against prostaglandin F_2α stopped cycling after one to three normal cycles. Laparotomy at 30 days after the last estrus revealed no recently formed corpora lutea. During the remaining 70 days of observation the antibody titer increased to 1:700, accompanied by increasing total serum estrogens (136 pg/ml at day 100) and a slow decline in circulating progesterone levels (0.6 ng/ml at day 100). The ovaries at day 100 contained degenerated corpora lutea and luteinized follicles. The suppression of the estrous cycle in the present experiments was interpreted as resulting from prolongation of luteal function as well as from inhibition of ovulation.     

The gelling of κ-carrageenan in light and heavy water

Gels of κ-carrageenan were prepared in H₂O and D₂O. Rheological measurements revealed that in heavy water the elastic modulus is substantially higher. From the rheological view point, this result indicates the formation of a higher number of physical-crosslinking points in the structure of the gel. These crosslinking points are mainly associated with the aggregation of the double-helix of the κ-carrageenan, which is enhanced in D₂O. The thermal stability of the gels in both solvents (relative to the sol-gel transition) was also investigated by using polarimetric measurements (to measure the relative fraction of helix-coil) and micro-differential scanning calorimetry (micro-DSC). The results obtained using these techniques are very consistent and allow us to propose a model to explain the differences for the gels formed in light and heavy water based on the degree of double-helix aggregation.     

Healthcare Utilization and Clinical Outcomes after Catheter Ablation of Atrial Flutter

Atrial flutter ablation is associated with a high rate of acute procedural success and symptom improvement. The relationship between ablation and other clinical outcomes has been limited to small studies primarily conducted at academic centers. We sought to determine if catheter ablation of atrial flutter is associated with reductions in healthcare utilization, atrial fibrillation, or stroke in a large, real world population. California Healthcare Cost and Utilization Project databases were used to identify patients undergoing atrial flutter ablation between 2005 and 2009. The adjusted association between atrial flutter ablation and healthcare utilization, atrial fibrillation, or stroke was investigated using Cox proportional hazards models. Among 33,004 patients with a diagnosis of atrial flutter observed for a median of 2.1 years, 2,733 (8.2%) underwent catheter ablation. Atrial flutter ablation significantly lowered the adjusted risk of inpatient hospitalization (HR 0.88, 95% CI 0.84–0.92, p<0.001), emergency department visits (HR 0.60, 95% CI 0.54–0.65, p<0.001), and overall hospital-based healthcare utilization (HR 0.94, 95% CI 0.90–0.98, p = 0.001). Atrial flutter ablation was also associated with a statistically significant 11% reduction in the adjusted hazard of atrial fibrillation (HR 0.89, 95% CI 0.81–0.97, p = 0.01). Risk of acute stroke was not significantly reduced after ablation (HR 1.09, 95% CI 0.81–1.45, p = 0.57). In a large, real world population, atrial flutter ablation was associated with significant reductions in hospital-based healthcare utilization and a reduced risk of atrial fibrillation. These findings support the early use of catheter ablation for the treatment of atrial flutter.     

A simple,reliable method to determine the mean gait speed using heel markers on a treadmill

Gait speed is an essential parameter of gait analysis. Our study proposed a simple and accurate method to extract a mean gait speed during walking on a treadmill using only kinematic data from markers placed on the heels of the participants’ feet. This method provided an attractive, simple method that remains resistant to errors in treadmill calibration. In addition, this method required only two markers, since heel markers are essential to gait analysis, and the proposed method is robust enough to differentiate among various gait speeds (mean error <1%).     

Comparative Genomic Analysis Reveals a Possible Novel Non-Tuberculous Mycobacterium Species with High Pathogenic Potential

Mycobacteria have been reported to cause a wide range of human diseases. We present the first whole-genome study of a Non-Tuberculous Mycobacterium, Mycobacterium sp. UM_CSW (referred to hereafter as UM_CSW), isolated from a patient diagnosed with bronchiectasis. Our data suggest that this clinical isolate is likely a novel mycobacterial species, supported by clear evidence from molecular phylogenetic, comparative genomic, ANI and AAI analyses. UM_CSW is closely related to the Mycobacterium avium complex. While it has characteristic features of an environmental bacterium, it also shows a high pathogenic potential with the presence of a wide variety of putative genes related to bacterial virulence and shares very similar pathogenomic profiles with the known pathogenic mycobacterial species. Thus, we conclude that this possible novel Mycobacterium species should be tightly monitored for its possible causative role in human infections.