Confocal laser scanning microscopy (CLSM) offers improved depth discrimination and spatial resolution to the analysis of biologic samples. We demonstrate in this paper that such technology is ...valuable in examining DNA single‐strand breaks in human cells. The single‐cell‐gel (SCG) assay is a new technique for measuring DNA strand breaks in individual cells. Cells embedded in lowmelting‐point agarose are treated with varying concentrations of hydrogen peroxide to induce DNA strand breaks. Following cell lysis and alkaline electrophoresis, which enables single‐stranded break detection, analysis of the resulting “comets” provides an accurate method of comparing changes in DNA migration patterns, which have been shown to reflect the DNA damage levels. A statistically significant difference (p < 0.01) in single‐stranded DNA damage levels was detected in cells exposed to hydrogen peroxide concentrations as low as 10 nm for 2 min. LSM analysis of the SCG technique allows rapid, sensitive and reproducible quantitation of single‐stranded breaks of cellular DNA.
The single-cell gel (SCG) assay has been shown to be a valuable technique to measure DNA-strand breaks in individual cells. Imaging of the assay is enhanced by laser-scanning microscopy (LSM) ...technology. Cells embedded in low melt point agarose were treated with 12.5 mM H2O2 to induce DNA-strand breaks. Following cell lysis and electrophoresis under alkaline conditions to allow single-stranded break detection, analysis of the resulting comets can provide an accurate method of comparing DNA-damage levels. The migration patterns of stained DNA were quantitated by LSM analysis. A statistically significant time-dependent dose-response relationship was clearly observed. LSM analysis of the SCG technique allows rapid, sensitive, and reproducible quantitation of single-stranded breaks and alkali-labile sites in the DNA of single cells.
SUMMARY
The Arabidopsis thaliana aldehyde oxidase 3 (AAO3) catalyzes the oxidation of abscisic aldehyde (ABal) to abscisic acid (ABA). Besides ABal, plants generate other aldehydes that can be toxic ...above a certain threshold. AAO3 knockout mutants (aao3) exhibited earlier senescence but equivalent relative water content compared with wild‐type (WT) during normal growth or upon application of UV‐C irradiation. Aldehyde profiling in leaves of 24‐day‐old plants revealed higher accumulation of acrolein, crotonaldehyde, 3Z‐hexenal, hexanal and acetaldehyde in aao3 mutants compared with WT leaves. Similarly, higher levels of acrolein, benzaldehyde, crotonaldehyde, propionaldehyde, trans‐2‐hexenal and acetaldehyde were accumulated in aao3 mutants upon UV‐C irradiation. Aldehydes application to plants hastened profuse senescence symptoms and higher accumulation of aldehydes, such as acrolein, benzaldehyde and 4‐hydroxy‐2‐nonenal, in aao3 mutant leaves as compared with WT. The senescence symptoms included greater decrease in chlorophyll content and increase in transcript expression of the early senescence marker genes, Senescence‐Related‐Gene1, Stay‐Green‐Protein2 as well as NAC‐LIKE, ACTIVATED‐BY AP3/P1. Notably, although aao3 had lower ABA content than WT, members of the ABA‐responding genes SnRKs were expressed at similar levels in aao3 and WT. Moreover, the other ABA‐deficient mutants aba2 and 9‐cis‐poxycarotenoid dioxygenase3‐2 (nced3‐2), that has functional AAO3 exhibited similar aldehydes accumulation and chlorophyll content like WT under normal growth conditions or UV‐C irradiation. These results indicate that the absence of AAO3 oxidation activity and not the lower ABA and its associated function is responsible for the earlier senescence symptoms in aao3 mutant.
Significance Statement
The current study demonstrates that AAO3, known to catalyze the last step of ABA biosynthesis by oxidizing ABal, is essential in preventing leaf damage by oxidizing toxic levels of aldehydes in rosette leaves. We provided credible evidences that premature senescence in this study was not because of low ABA content and its consequential function in aao3, but rather due to higher aldehyde accumulation, which is caused by AAO3 deficiency.
Abstract Since 1990, a wide range of palliative care services has been implemented throughout the Catalan Health Care System. In 2005, 21,400 patients received palliative care; 59% had cancer (79.4% ...of all cancer patients) and 41% had other noncancer diagnoses (25.0%–56.5% of all noncancer patients). Today, more than 95% of Catalonia is covered by palliative care services. Fourteen districts have comprehensive palliative care networks. A total of 140 full-time physicians work in 183 specialty programs, including 63 palliative care units (with a total of 552 beds), 34 hospital consult teams, 70 home care teams, 16 outpatient clinics, and specialized pediatric and HIV/AIDS consult teams. Opioid consumption increased from 3.5 mg per capita in 1989 to 21 mg per capita population in 2004. The cost of the specialist palliative care network is more than 40 million Euros annually. However, the cost efficiency is striking. Due to the radical change in the use of acute and emergency beds, the project saves the Catalan Health Care System an estimated 48 million Euros annually, a net savings of 8 million Euros annually. Additional preliminary data suggest that symptom control and patient/family satisfaction are both improved by these services.
Summary A post-Millennium Development Goals agenda for health in Bangladesh should be defined to encourage a second generation of health-system innovations under the clarion call of universal health ...coverage. This agenda should draw on the experience of the first generation of innovations that underlie the country's impressive health achievements and creatively address future health challenges. Central to the reform process will be the development of a multipronged strategic approach that: responds to existing demands in a way that assures affordable, equitable, high-quality health care from a pluralistic health system; anticipates health-care needs in a period of rapid health and social transition; and addresses underlying structural issues that otherwise might hamper progress. A pragmatic reform agenda for achieving universal health coverage in Bangladesh should include development of a long-term national human resources policy and action plan, establishment of a national insurance system, building of an interoperable electronic health information system, investment to strengthen the capacity of the Ministry of Health and Family Welfare, and creation of a supraministerial council on health. Greater political, financial, and technical investment to implement this reform agenda offers the prospect of a stronger, more resilient, sustainable, and equitable health system.
Background. The risk of overt and subtle cerebral injury may be higher in intracardiac operation (ICO) rather than coronary artery bypass grafting (CABG). S-100 protein is a specific astroglial ...protein whose serum level increases after cerebral injury. Elevated serum levels of S-100 have been detected after adult cardiac operations and correlated with neurologic injury.
Methods. The level of S-100 protein was measured serially over 24 hours in 40 patients (27 undergoing aortic valve replacement, 9 mitral valve replacement, 4 closure of atrial septal defect) undergoing ICO and 20 patients undergoing CABG.
Results. The groups were similar with respect to age and cardiopulmonary bypass times. The S-100 level was not elevated before operation in any patient. Peak S-100 levels were reached at skin closure, when 35 of the ICO patients (88%) and 13 of the CABG patients (65%) had elevated S-100 levels. At skin closure peak S-100 levels were significantly greater in the ICO group (median interquartile range, 0.76 0.44–1.16 versus 0.3 0–0.55 μg/L; p < 0.01). At 5 hours S-100 levels were still elevated in 22 patients in the ICO group compared with 1 patient in the CABG group (p < 0.01), and at 24 hours 17 ICO patients had persistently elevated S-100 levels in comparison with 2 in the CABG group (p < 0.01). One valve patient had a stroke 24 hours after operation accompanied by a secondary increase in the S-100 level. There was no significant difference in postoperative S-100 levels between 5 patients in the ICO group with a prior history of stroke and those without. The peak S-100 level correlated with patient age (r = 0.59; p < 0.001) but not with the duration of cardiopulmonary bypass or core temperature during the operation.
Conclusions. Intracardiac operation results in a significantly greater elevation in S-100 levels than CABG. Elevated S-100 levels correlate with increasing patient age but not with the duration of cardiopulmonary bypass or intraoperative core temperature. These findings raise the possibility that ICO patients may be more vulnerable to even subtle levels of cerebral injury than CABG patients.
(Ann Thorac Surg 1997;63:492–6)
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