An abrupt increase in foliage ABA levels on incipient leaf death occurs across vascular plants.

Forest mortality during drought has been attributed to hydraulic failure, which can be challenging to measure. A limited number of alternative proxies for incipient leaf death exist. Here we investigate whether a terminal increase in abscisic acid (ABA) levels in leaves occurs across vascular land plants and is an indicator of imminent leaf death. For different species across vascular plants, we monitored ABA levels during lethal drought as well as leaf embolism resistance, across the canopy as leaves die following senescence, or when leaves are exposed to a heavy, lethal frost late in the growing season. We observed a considerable increase in foliage ABA levels once leaves showed signs of incipient leaf death. This increase in ABA levels upon incipient leaf death, could be induced by embolism during drought, by freezing or as leaves age naturally, and was observed in species spanning the phylogeny of vascular land plants as well as in an ABA biosynthetic mutant plant. A considerable increase in foliage ABA levels may act as an indicator of impending leaf death.

Indoor NO2 air pollution and lung function of professional cooks.

Studies of cooking-generated NO2 effects are rare in occupational epidemiology. In the present study, we evaluated the lung function of professional cooks exposed to NO2 in hospital kitchens. We performed spirometry in 37 cooks working in four hospital kitchens and estimated the predicted FVC, FEV1 and FEF(25-75), based on age, sex, race, weight, and height, according to Knudson standards. NO2 measurements were obtained for 4 consecutive days during 4 different periods at 20-day intervals in each kitchen. Measurements were performed inside and outside the kitchens, simultaneously using Palm diffusion tubes. A time/exposure indicator was defined as representative of the cumulative exposure of each cook. No statistically significant effect of NO2 exposure on FVC was found. Each year of work as a cook corresponded to a decrease in predicted FEV1 of 2.5% (P = 0.046) for the group as a whole. When smoking status and asthma were included in the analysis the effect of time/exposure decreased about 10% and lost statistical significance. On predicted FEF(25-75), a decrease of 3.5% (P = 0.035) was observed for the same group and the inclusion of controllers for smoking status and asthma did not affect the effects of time/exposure on pulmonary function parameter. After a 10-year period of work as cooks the participants of the study may present decreases in both predicted FEV1 and FEF(25-75) that can reach 20 and 30%, respectively. The present study showed small but statistically significant adverse effects of gas stove exposure on the lung function of professional cooks.

Rapid, reliable and inexpensive quality assessment of biotinylated cRNA.

The interpretation of oligonucleotide array experiments depends on the quality of the target cRNA used. cRNA target quality is assessed by quantitative analysis of the representation of 5' and 3' sequences of control genes using commercially available Test arrays. The Test array provides an economically priced means of determining the quality of labeled target prior to analysis on whole genome expression arrays. This manuscript validates the use of a duplex RT-PCR assay as a faster (6 h) and less expensive (<10 US dollars) but equally accurate alternative to the Test arrays in determining biotinylated cRNA quality. Forty-one different cRNA samples were hybridized to HG-U133A microarrays from Affymetrix. Ten cRNA samples with a beta-actin 3'/5' ratio >6 were chosen and classified as degraded cRNAs, and 31 samples with a beta-actin 3'/5' ratio <6 were selected as good quality cRNAs. Blinded samples were then used for the RT-PCR assay. After gel electrophoresis, optical densities of the amplified 3' and 5' fragments of beta-actin were measured and the 3'/5' ratio was calculated. There was a strong correlation (r(2) = 0.6802) between the array and the RT-PCR beta-actin 3'/5' ratios. Moreover, the RT-PCR 3'/5' ratio was significantly different (P < 0.0001) between undegraded (mean +/- SD, 0.34 +/- 0.09) and degraded (1.71 +/- 0.83) samples. None of the other parameters analyzed, such as i) the starting amount of RNA, ii) RNA quality assessed using the Bioanalyzer Chip technology, or iii) the concentration and OD260/OD280 ratio of the purified biotinylated cRNA, correlated with cRNA quality.

Immunotherapy with acute leukemia cells modified into antigen-presenting cells: ex vivo culture and gene transfer methods.

Adult patients with acute leukemia have, in general, a poor prognosis, with long-term, disease-free survival achieved in only approximately one-third of cases. One of the proposed mechanisms for this poor overall response is the inability of the immune system to detect and eliminate residual malignant leukemia cells, which subsequently serve as a source of leukemic relapse. This review discusses the rationale of immunotherapy for acute leukemia and presents in vitro and in vivo model systems that were devised for pre-B acute lymphocytic leukemia (ALL) and acute myeloid leukemia (AML). New advances in the ex vivo manipulation of acute leukemia cells are presented, which attempt to modify these cells into functional antigen-presenting cells. These cells can then be used as autologous vaccines at the time of minimal residual disease after standard chemotherapy, to stimulate host immune responses against their own leukemia cells. The various approaches toward this aim include incubation of leukemia cells with cytokines or growth factors and gene manipulation of these cells. In particular, ex vivo culture of ALL cells with CD40 ligand, incubation of AML cells with granulocyte-macrophage colony-stimulating factor and interleukin-4 (GM-CSF/IL-4) and lentiviral transduction of ALL and AML cells for expression of immunomodulators (CD80 and GM-CSF) are current approaches under investigation for the development of autologous acute leukemia cell vaccines.

Biologic response modifiers in acute lymphoblastic leukemia.

Despite the presence of tumor antigens, the paucity of clinically significant T-cell mediated immune responses against human tumors is striking. This may, in part, be because of the inability of cancer cells to function as efficient antigen-presenting cells. For full activation, T cells must receive two signals delivered by antigen-presenting cells. The first is antigen-specific and is delivered by presentation of antigenic peptide by the major histocompatibility complex molecules to the T-cell receptor. This signal, although necessary, is in itself insufficient to mediate T-cell activation, cytokine release, and subsequent T-cell proliferation and function. For full T-cell activation, T cells require delivery of a secondary, costimulatory signal, such as that delivered by members of the B7 family to their receptor on the T-cell, CD28. Delivery of an antigen signal in the absence of costimulation does not result in productive immunity, but rather in anergy, a state of antigen-specific T-cell nonresponsiveness. To induce T-cell proliferation against B-cell malignancies, the tumor cell must first be induced to express B7 or the tumor antigen must be presented by an efficient antigen-presenting cell. Simple expression of B7 on the tumor cell alone, however, cannot reverse anergy. Reversal of anergy is a complex process involving stepwise repair of the T-cell defect and can be accomplished by prolonged exposure to interleukin-2, signaling through the CD2 pathway, followed by antigen presentation with B7-mediated costimulation. Successful immunotherapeutic strategies in the B-cell malignancies will likely require steps to reverse established anergy in the tumor-bearing host as well as effective tumor-antigen presentation.

Additive effects of steel factor and antisense TGF-beta 1 oligodeoxynucleotide on CD34+ hematopoietic progenitor cells.

We previously demonstrated that TGF-beta 1 antisense oligodeoxynucleotides can release early CD34+ bone marrow progenitors from quiescence, and increase the numbers of mixed and large erythroid colonies. As Steel Factor (SF) has a similar effect on colony formation by CD34+ cells, we tested whether this factor acts by blocking the inhibitory effects of TGF-beta. That this is not generally the case was demonstrated by the finding that the combination of TGF-beta 1 antisense with SF in cultures of CD34+ bone marrow cells yielded enhanced colony formation that was more than additive when compared to cultures containing the single agents. This combination also yielded enhanced colony formation by CD34+ umbilical cord blood cells, but in this case, the effect was slightly less than additive. Thus in cord blood, some, but not all, of the progenitors that are maintained in quiescence by TGF-beta can be triggered into cycle by SF. However, the absolute number of CFU-GEMMs found in the antisense TGF-beta plus SF cultures of cord blood was 4-fold higher than that found in comparable bone marrow cultures. These data correlate well with our previous observations that umbilical cord blood contains 4-fold more CD34+ CD38- cells, the population found to respond to TGF-beta 1 antisense oligodeoxynucleotides.

An improved panning technique for the selection of CD34+ human bone marrow hematopoietic cells with high recovery of early progenitors.

The human hematopoietic pluripotent repopulating "stem cell" is thought to be present within a minor subpopulation of CD34+ cells. This has not been definitively shown, although the more primitive CD34+ cell subset contains precursors for all lymphoid and nonlymphoid cell lineages. When purifying CD34+ cells, it is important to recover these early progenitors, which are more strongly immunoadsorbent to the separation devices. Using a commercialized panning system (AIS CELLector flasks), we observed that a high degree of purity requires a thorough washing procedure so that cells not binding or weakly binding to CD34 antibodies are removed. High recoveries can be obtained if the adherent cells are then efficiently detached by a 2-hour incubation in culture medium without added cytokines. In this way, we can routinely obtain 93.5 +/- 3.4% purity of CD34+ cells with a 74% yield of the multipotent colony-forming units (CFU-GEMM). Complete recovery of the putative "stem cell," or at least the early progenitor cell compartment (CD34+ CD38low/- CD34+ Thy-1+ cells), is also obtained. More than 30% of these cells can generate day-14 colonies in vitro. Comparable results were obtained when the separation was scaled up for clinical application using appropriate large-scale devices. The various incubation times of the procedure can be easily adjusted to the work schedule. This renders the procedure easy to handle, efficient, safe, and, because the cells can be observed under light microscopy, easy to control.

Fluorometric fiber optic drop sensor for atmospheric hydrogen sulfide.

A fluorometric technique based on a liquid drop excited from its interior by an optical fiber is described for the measurement of low concentrations of atmospheric hydrogen sulfide (H(2)S). A drop of alkaline fluorescein mercuric acetate (FMA) solution is suspended in a flowing air sample stream and serves as a renewable sensor. An optical fiber contained within the conduit that forms the drop, brings in the excitation beam; the fluorescence emission is measured by an inexpensive photodiode positioned close to the drop. As H(2)S in the sample is collected by the alkaline drop, it reacts rapidly with FMA resulting in a significant decrease in fluorescence intensity, proportional to the concentration of H(2)S sampled. The chemistry of this uniquely selective reaction has been well established for many years; the present technique permits a simple fast inexpensive near real-time measurement with very little reagent consumption. Even without prolonged sampling/preconcentration steps, limits of detection (LODs) in the double digit ppbv range is readily attainable.

Spectrophotometric detection of arsenic using flow-injection hydride generation following sorbent extraction preconcentration.

An automated system with a C(18) bonded silica gel packed minicolumn is proposed for spectrophotometric detection of arsenic using flow-injection hydride generation following sorbent extraction preconcentration. Complexes formed between arsenic(III) and ammonium diethyl dithiophosphate (ADDP) are retained on a C(18) sorbent. The eluted As-DDP complexes are merged with a 1.5% (w/v) NaBH(4) and the resulting solution is thereafter injected into the hydride generator/gas-liquid separator. The arsine generated is carried out by a stream of N(2) and trapped in an alkaline iodine solution in which the analyte is determined by the arsenomolybdenum blue method. With preconcentration time of 120 s, calibration in the 5.00-50.0 mug As l(-1) range and sampling rate of about 20 samples h(-1) are achieved, corresponding to 36 mg ADDP plus 36 mg ammonium heptamolybdate plus 7 mg hydrazine sulfate plus 0.7 mg stannous chloride and about 7 ml sample consumed per determination. The detection limit is 0.06 mug l(-1) and the relative standard deviation (n=12) for a typical 17.0 mug As l(-1) sample is ca. 6%. The accuracy was checked for arsenic determination in plant materials from the NIST (1572 citrus leaves; 1573 tomato leaves) and the results were in agreement with the certified values at 95% confidence level. Good recoveries (94-104%) of spiked tap waters, sugars and synthetic mixtures of trivalent and pentavalent arsenic were also found.

Kupffer cell activation with BCG. Corynebacterium parvum or zymosan protects against acute liver injury induced by carbon tetrachloride in rats.

Kupffer cells have been implicated in the pathogenesis of liver injury, but there is controversy about the effects of activation of these cells on the hepatotoxicity of chemicals and endotoxin. It has been shown that injection of Corynebacterium parvum in rats induces macrophage activation that protects against toxic effects of carbon tetrachloride and acetaminophen, five days after injection, and this protection is due to inhibition of microsomal oxidizing enzymes and increased production of glutathion. To verify if the protective effect occurs soon after Kupffer cell activation, with different activators, male albino rats were treated with intravenous injection of BCG (0.5 ml with 7.5.10(8) bacilli), Corynebacterium parvum (30 mg/kg) or zymosan (7.5.10(6) yeast cells). Fourty-eight hours after the injection of one of the macrophage activators, the animals and rats treated with intravenous injection of saline (controls) received carbon tetrachloride by subcutaneous route (1 ml/kg of CCl4, 3:1 in soybean oil). Fourty-eight hours after the animals were killed after ether anesthesia and fragments of the liver were fixed, paraffin embedded and the sections stained with hematoxylin and eosin. A Weibel grid with 168 points was used to estimate the percent volume of necrosis and severe hydropic degeneration. The results showed that the volume density of necrosis and severe hydropic degeneration were significatively lesser in rats treated with the three Kupffer cells activators. The protection was greater with BCG and Corynebacterium parvum than with zymosan. These results confirm that activation of Kupffer cell with three different activators can induce protection against liver cell injury produced by carbon tetrachloride in rats soon as 48 h after injection of activators.

[Magnifying colonoscopy in the diagnosis of colorectal carcinoma invading the submucosa in familial adenomatous polyposis]. / Colonoscopia com magnificação de imagem no diagnóstico de carcinoma colorretal invasivo da submucosa na polipose adenomatosa familiar.

The development of colonoscopy with image magnification has enable to study the colonic mucosa in detail and to do differential diagnosis between neoplastic and non-neoplastic lesions from the observation of pit patterns. The results are comparable to stereomicroscopy being possible to predict the histologic diagnosis. In a patient with familial adenomatous polyposis magnifying colonoscopy was performed and this method demonstrated a wide variation of benign polypoid lesions and the morphological features of early colorectal cancer. In this patient, the evaluation by image magnification, together with indigo carmin 0.4% chromoscopy, showed a wide variety of lesions in the colon and rectum: laterally spreading tumor in the cecum, with IIIL + IV pits, subpediculate polyp in the transverse colon with approximately 2.0 cm diameter and IV + V pits, flat elevated lesions IIIL type, and in the sigmoid colon IIa + IIc lesion with V type of Kudo's classification were observed. The evaluation of pit patterns of the lesions in the transverse and sigmoid colon has enable to do the endoscopic diagnosis of the lesion with submucosal invasion.

[Risk stratification in patients recuperating from acute myocardial infarct. How useful is it?]. / Estratificação do risco no doente que recuperou do enfarte agudo do miocárdio. Que utilidade?

The author intended to discuss the individual prognostic value of some best known risk stratification variables after Acute Myocardial Infarction (AMI), whose tests have a low diagnostic accuracy due to high risk individual variations, emphasizing the statistic errors of using one single risk variable on post AMI risk stratification schemes. Some epidemiologic data about AMI survivors are reviewed and the author concludes by emphasizing the importance of using more AMI population characteristics on post AMI risk stratification schemes.

[Who should be treated? How should they be treated? What has changed in arterial hypertension therapy?]. / Quem tratar? Como tratar? O que mudou na terapêutica da HTA.

The author briefly revises the paradoxical changes that occurred in hypertension treatment, according the answers to the two following questions: Who to treat? How to treat? Emphasizes the main importance of non-drug treatment for the preservation of the hypertensive's quality of life and hyperadrenergic responses control. Concludes that the hypertension treatment is so individualized as conditionated by a lot of factors (drug efficacity in lowering blood pressure levels, concomitant risk factors and cardiovascular diseases, own patient's adherence or preference, etc.).

[Calculation of cardiac output using Doppler echocardiography]. / Cálculo do débito cardíaco por ecocardiografia Doppler.

The authors describe the theoretical basis and general methods for calculating Cardiac Output (CO) by Doppler Echocardiography, using triangulation and planimetric methods ro obtain the Average Velocity (AV) of the flows. They point the more advisable Bidimensional Echocardiography views plans for the measurement of aortic, mitral, pulmonary and tricuspid cross sectional areas (CA) used in calculating the respective outputs (CO = CA x AV x cardiac frequency) and the rate Pulmonary Flow/Systemic Flow (PF/SF). They point out the main technological limitations, alternative methods and places for calculating CO in certain situations (Aortic stenosis, Pulmonary stenosis, etc.). They stress that these are alternative methods to the invasive techniques in the clinic Cardiac Pathologies (infants and adults), many of them treated in Intensive Care units, where the prolonged non invasive monitorization of CO is facilitated by the use of the same CA.

[Prognostic significance of angina pectoris before myocardial infarction]. / Significado prognóstico da angina de peito prévia ao enfarte do miocárdio.

The AA analysed the incidence of previous angina to myocardial infarction (PA) (42.3%) and post myocardial infarction angina (PMIA) (46.39%) in 97 patients that survived the acute phase of myocardial infarction, all discharged from the CCU of the Funchal's Hospital Center Cardiological Department (Madeira Island), whose 25 of them (26.8%) presented both. They met 14 positive Treadmill tests in the 30 patients that were submitted to sub-maximal protocols. They concluded that the presence of PA and positive treadmill tests can identify a patient group with increased risk of PMIA, suggesting that patients with PA have also an increased isquemic risk.

[Assessment of the first myocardial infarctions with lead ECG --implications in the study of hospital mortality and its reduction using thrombolytic drugs]. / Avaliação da dimensão dos primeiros enfartes do miocárdio pelo ECG de superfície--implicações no estudo da mortalidade intra-hospitalar e sua redução por fármacos trombolíticos.

STUDY OBJECTIVE: Correlation between mortality reduction of first Myocardial Infarction (MI) by thrombolytic therapy and MI size evaluated with the classical Electrocardiogram (ECG). DESIGN: A retrospective sequential study. SETTING: Coronary Unit patients. PATIENTS: Sequential sample of 132 patients with first MI obeying all the following criteria: 1) no previous MI; 2) age less than or equal to 70 years; 3) clinical evolution less than 12 hours; 4) no Left Bundle Branch Block in the CCU first ECG; 5) ischemic ST elevation in greater than or equal to 1 initial ECG leads. Patients were divided into Group A, with less than or equal to 3 initial ECG leads with ischemic ST elevation (n = 80), and Group B, with greater than or equal to 4 initial ECG leads with ischemic ST elevation (n = 52). Only 34 patients (25.7%) did thrombolytic therapy with IV Streptokinase (SK); 15 from Group A and 19 from Group B. MEASUREMENTS AND MAIN RESULTS: 17 patients died in MI acute phase (12.8%); 4 in Group A (5%) and 13 in Group B (24.9%). Inhospital mortality was statistically worst in Group B than in Group A (24.9% vs 5% with p less than 0.01). Creatin kinase (CK) maximal values (A = 911.5 UI; B = 1444.6 UI with p less than 0.01) and initial Heart Rate (A = 75.7; B = 86.7 with p less than 0.001) were also statistically greatest in Group B. Inhospital mortality was smaller in patients treated with SK (8.8% vs 14.3%), as in Group B (10.5% vs. 33.3%), both without statistical significance. CONCLUSIONS: Inhospital mortality and thrombolytic therapy benefit were so bigger as MI size evaluated by the number of initial ECG leads with ischemic ST elevation, by initial HR and maximal values of CK. Classical ECG can be useful by identifying patients with first MI that can more benefit with thrombolytic therapy (greater than or equal to 4 leads with ischemic ST elevation).

Mortality and morbidity trends in ischemic heart disease in the autonomous region of Madeira in the ten-year period 1987-1996.

The increase in absolute number of deaths from ischemic heart disease (IHD) in the population aged > or = 65 years, in both sexes, in Madeira, when comparing the years 1987 and 1996, led to significant increases in the corresponding standardized death rates that go against the stabilization seen at national level. Significant increases in these rates for the same years were also seen in the district of Beja and in the Azores. The aim of this study was to ascertain the trends for the incidence, morbidity and mortality from acute myocardial infarction (AMI) in patients admitted in Madeira and its contribution to the increase in these rates, particularly in the population aged < 65 years of both sexes, which the number of deaths from ischemic heart disease did not increase. We studied 119 pts with AMI admitted in 1987 (year A), of whom 53 were aged < 65 years, and 186 pts with AMI admitted in 1996 (year B), of whom 72 were aged < 65 years, whose data were included in the Madeira Ischemic Heart Disease Register (RECIMA), an IHD hospital register that covers 1792 patients admitted with AMI in the Coronary Intensive Care Unit of the Department of Medical and Surgical Cardiology of Funchal Hospital over a period of 15 years (1984-1998). Mortality by the 28th day (fatal AMI admissions) in all ages fell slightly in both sexes in the two years studied (A = 19.3%; B = 16.1%). The number of fatal AMI admissions rose among females in the two age groups considered A = 11; B = 20; delta% = +45) and fell among males (A = 12; B = 10; delta% = -20). In males aged > or = 65 years, this number remained the same (A = 7; B = 7) and fell in males aged > or = 65 years (A = 5; B = 3; delta% = -40). The number of pts who survived to the 28th day (non-fatal AMI admissions) rose in all age groups for both sexes (A = 96; B = 156; delta% = +38.46), as did the ratios with deaths from IHD. These increases were roughly double in the group of patients aged 65 years compared to patients aged < 65 years. We found highly significant positive correlations in the population aged < 65 years between the number of non-fatal AMI admissions (morbidity data) and the number of deaths from IHD (mortality data) recorded in every year of the 10-year period 1987-96, these values being highly significant in both sexes (r = 0.89; p < 0.0001), in males (r = 0.87; p < 0.0001) and in females (r = 0.77; p < 0.0001). Since our study was carried out on an island on which all AMI cases are admitted to a single treatment center, we can conclude that these positive correlations represent a trend towards worsening of morbidity and mortality from IHD in Madeira in the population aged < 65 years, even though the number of deaths from IHD did not rise. The establishment of IHD registers similar to RECIMA in other regions of the country would help to identify trends in morbidity, mortality, and morbidity plus mortality in this population that would be useful in improving the orientation of resources allocated to the prevention and treatment of cardiovascular diseases.

Activities and participation in children with developmental coordination disorder: a systematic review.

PURPOSE: To systematically review all literature published in peer reviewed journals from January 1995 to July 2008 in order to summarize and describe the activity limitations and participation restrictions of children with developmental coordination disorder (DCD). METHODS: Multiple databases were systematically searched for articles related to DCD; only descriptive, intervention or qualitative articles were retained. Articles were coded using the International Classification of Function, Disability and Health (ICF) and descriptions of the activity and participation issues of individuals with DCD were identified. RESULTS: Data analysis revealed that, from 371 articles that met inclusion criteria, only 44 (14.4%) presented any data related to activity or participation issues. Information was inconsistent and only 18 articles used published measurement tools. Most frequently cited issues were poor handwriting, difficulties playing ball games, getting dressed and participating in organized sports. CONCLUSION: Evidence concerning activity and participation issues for children with DCD is limited in both volume and scope. Improved understanding of participation and of activity limitations in children with DCD is essential for clarifying diagnostic criteria, guiding assessment, and making evidence-based decisions regarding intervention. Researchers working with this population should make every effort to measure and consistently report the impact of children's motor impairments on function.

Measurements and modeling of reactive nitrogen deposition in southeast Brazil.

Increased reactive nitrogen (Nr) deposition due to expansion of agro-industry was investigated considering emission sources, atmospheric transport and chemical reactions. Measurements of the main inorganic nitrogen species (NO2, NH3, HNO3, and aerosol nitrate and ammonium) were made over a period of one year at six sites distributed across an area of ∼130,000 km2 in southeast Brazil. Oxidized species were estimated to account for ∼90% of dry deposited Nr, due to the region's large emissions of nitrogen oxides from biomass burning and road transport. NO2-N was important closer to urban areas, however overall HNO3-N represented the largest component of dry deposited Nr. A simple mathematical modeling procedure was developed to enable estimates of total Nr dry deposition to be made from knowledge of NO2 concentrations. The technique, whose accuracy here ranged from <1% to 29%, provides a useful new tool for the mapping of reactive nitrogen deposition.