Stratification of Patients with Chronic Obstructive Pulmonary Disease Using Volatile Organic Compounds.

Chronic obstructive pulmonary disease (COPD) is a common disease that causes long-term disability and death. Its natural history is punctuated by acute worsening of symptoms, called exacerbations, which are associated with increased mortality and hospitalization. In this work, we aim to stratify patients with COPD based on their risk for exacerbation; for this purpose, we employ non-invasive biomarkers, that is, volatile organic compounds (VOCs), acquired from the patients' exhaled breath coupled with their spirometry and age. We utilize a series of classification schemes with the best performing one achieving overall Accuracy = 93.5%. The yielded results are, therefore, encouraging and prompt for further investigation toward the utilization of VOCs in the management of COPD.

Reducing MCM levels in human primary T cells during the G(0)-->G(1) transition causes genomic instability during the first cell cycle.

DNA replication is tightly regulated, but paradoxically there is reported to be an excess of MCM DNA replication proteins over the number of replication origins. Here, we show that MCM levels in primary human T cells are induced during the G(0)-->G(1) transition and are not in excess in proliferating cells. The level of induction is critical as we show that a 50% reduction leads to increased centromere separation, premature chromatid separation (PCS) and gross chromosomal abnormalities typical of genomic instability syndromes. We investigated the mechanisms involved and show that a reduction in MCM levels causes dose-dependent DNA damage involving activation of ATR & ATM and Chk1 & Chk2. There is increased DNA mis-repair by non-homologous end joining (NHEJ) and both NHEJ and homologous recombination are necessary for Mcm7-depleted cells to progress to metaphase. Therefore, a simple reduction in MCM loading onto DNA, which occurs in cancers as a result of aberrant cell cycle control, is sufficient to cause PCS and gross genomic instability within one cell cycle.

CDKN2B methylation status and isolated chromosome 7 abnormalities predict responses to treatment with 5-azacytidine.

5-Azacytidine, a DNA methyl transferase inhibitor, is effective in patients with myelodysplastic syndromes (MDS). Whether responses to 5-Azacytidine are achieved by demethylation of key genes or by cytotoxicity is unclear. Of 34 patients with MDS or acute myeloid leukaemia (AML) treated with 5-Azacytidine, 7 achieved complete remissions (CR) (21%) and 6 achieved haematological improvement. All six had less than 5% bone marrow (BM) blasts at the time of haematological improvements (HI) (2 had pre-existing refractory anaemia (RA), 4 had refractory anaemia with excess blasts (RAEB)). A further patient with RAEB had blast reduction to less than 5% without HI. Five of the seven (71%) complete responders had chromosome 7 abnormalities. BM CR predicted longer overall survival (OS) (median 23 versus 9 months, P=0.015). Bisulphite genomic sequencing (BGS) of the CDKN2B (p15(INK4b)) promoter showed low level, heterogeneous pretreatment methylation (mean 12.2%) in 14/17 (82%) patients analysed. Lower baseline methylation occurred in responders (9.8% versus 16.2% in non-responders P=0.07). No response was seen in patients with >24% methylation, in whom p15(INK4b) mRNA was not expressed. 5-Azacytidine reduced CDKN2B methylation by mean 6.8% in 8/17 (47%) patients, but this did not correlate with response. At 75 mg/m(2), cell death (reduced BM cellularity (P=0.001) and increased apoptosis (P=0.02)) rather than demethylation of CDKN2B correlates with response. Patients with >24% methylation may benefit from alternative dosing or combination strategies.

Static stabilometry, transcranial Doppler, and single photon emission computed tomography in patients with central dizziness.

Previous studies have found that transcranial doppler (TCD) and single photon emission computed tomography (SPECT) are effective means of diagnosing cerebral blood flow disorders in patients with central dizziness whose etiology was unknown by standard audiologic and/or vestibular assessment techniques. Also, static stabilometry, which measures a person's standing center of pressure (COP) movements, has been used to distinguish between patients with central neurologic and peripheral vestibular disorders. The purpose of this retrospective study was to examine the relation between TCD, SPECT, and stabilometry in patients with central dizziness attributable to cerebral blood flow disorders. Stabilometry testing was conducted on 50 normal subjects and 31 subjects with dizziness, the latter group consisting of persons with cerebral dysautoregulation, migraines, and unknown etiology with negative or positive SPECT results. The results indicated that patients with cerebral dysautoregulation were not significantly different from normal subjects or the other three groups in their COP movements. The other three groups exhibited significantly higher COP movements than the normal subjects, particularly when visual inputs were compromised. Patients with negative SPECT results were significantly different in their COP movements from the other three groups of subjects with dizziness. These results suggest that the pattern of COP movements may be useful in identifying patients with postural dysfunctions whose etiology may then be detected by TCD and SPECT.

Dihydropyridine-sensitive Ca2+ spikes and Ca2+ currents in rabbit ciliary body epithelial cells.

Intracellular microelectrode and whole-cell patch-clamp recordings were used to investigate a Ba(2+)-induced regenerative depolarization and its underlying Ba2+ current in the ciliary body epithelial cells of the rabbit eye. Exposure of these epithelial cells to 4-10 mmol l-1 Ba2+ depolarized the membrane potential and caused the generation of one or more spikes, before the membrane potential reached a steady-state level. The spikes, but not the slow phase of depolarization, could be blocked with Co2+ (2 mmol l-1), Gd3+ (25 mumol l-1), La3+ (20 mumol l-1), Cd2+ (10 mumol l-1), verapamil (30 mumol l-1) and nifedipine (1 mumol l-1). Tetrodotoxin at 100 nmol l-1 had no effect. In the absence of Na+, but in the presence of external Ba2+, step depolarization of the membrane potential activated an inward current that could be blocked with Co2+ (2 mmol l-1), Cd2+ (10 mumol l-1) and nifedipine (1 mumol l-1), but not with Ni2+ (50 mumol l-1) or omega-conotoxin (1-10 mumol). This inward current could be enhanced with the dihydropyridine agonist (+/-)BAY K 8644 (1 mumol l-1). The inactivation characteristics of the inward current (v1/2 = -38.7 mV, k = 12.6 mV) is most like that seen in neurons. These findings indicate that the epithelial cells of the ciliary body possess dihydropyridine-sensitive, voltage-activated Ca2+ channels.