Sap flow of sweet cherry reveals distinct effects of humidity and wind under rain covered and netted protected cropping systems.

Protected cropping systems (PCS) alter the plant growing environment, though understanding of this in ventilated systems and how the new climate affects tree water uptake is limited. Sap flow sensors and weather stations were deployed in 16-year-old 'Lapins' on 'Colt' rootstock cherry trees under a ventilated Voen PCS and in an adjacent bird netted PCS. Average and maximum temperatures were consistently higher (14.7 °C and 22.9 °C) while total daily solar radiation and average wind were consistently lower (12.9 MJ/m2 and 0.2 m/s) in rain covered, in contrast to netted, PCS (13.9 °C, 21.3 °C, 13.7 MJ/m2 and 0.9 m/s). Over the season, a threefold lower daily sap flow rate was observed under rain covered PCS. Using generalised additive modelling (GAM), the influence of individual climate parameters on sap flow were predicted. Whilst sap flow was only slightly affected by relative humidity (RH) less than 60%, above this threshold sap flow rapidly declined under rain covered PCS whereas sap flow more gradually declined above 20% RH under netted PCS. Overall, our novel modelling approach led to the discovery of the 60% RH critical threshold on predicted sap flow and the indirect effect that wind speeds have on sap flow under PCS.

HLA genotyping in the clinical laboratory: comparison of next-generation sequencing methods.

Implementation of human leukocyte antigen (HLA) genotyping by next-generation sequencing (NGS) in the clinical lab brings new challenges to the laboratories performing this testing. With the advent of commercially available HLA-NGS typing kits, labs must make numerous decisions concerning capital equipment and address labor considerations. Therefore, careful and unbiased evaluation of available methods is imperative. In this report, we compared our in-house developed HLA NGS typing with two commercially available kits from Illumina and Omixon using 10 International Histocompatibility Working Group (IHWG) and 36 clinical samples. Although all three methods employ long range polymerase chain reaction (PCR) and have been developed on the Illumina MiSeq platform, the methodologies for library preparation show significant variations. There was 100% typing concordance between all three methods at the first field when a HLA type could be assigned. Overall, HLA typing by NGS using in-house or commercially available methods is now feasible in clinical laboratories. However, technical variables such as hands-on time and indexing strategies are sufficiently different among these approaches to impact the workflow of the clinical laboratory.

Yeast strain affects phenolic concentration in Pinot noir wines made by microwave maceration with early pressing.

AIMS: This study examined the effects of yeast strains in a novel winemaking process that had been designed to optimize phenolic extraction and improve production efficiency for Pinot noir winemaking. METHODS AND RESULTS: Microwave maceration with early pressing and co-inoculation of yeast and malolactic bacteria for simultaneous alcoholic and malolactic fermentation was investigated. Yeast treatments (Saccharomyces cerevisiae RC212 and EC1118, and Saccharomyces bayanus AWRI1176) were co-inoculated with Oenococcus oeni PN4 immediately after must microwave maceration. Alcoholic and malolactic fermentation were complete 17 days postinoculation for all three yeast treatments. At 16-month bottle age, the AWRI1176-treated wines had approximately twice the nonbleachable pigment and colour density of wines fermented by EC1118 and RC212. CONCLUSIONS: The novel winemaking process produced Pinot noir wine that was stable 37 days after fruit had been harvested and yeast strain choice significantly impacted the stability and phenolic character of wine. SIGNIFICANCE AND IMPACT OF STUDY: Successful simultaneous alcoholic and malolactic fermentation in 17 days, and a demonstrated lack of inhibition between the yeast strains and malolactic strain applied in this study, provide proof of concept for very rapid red winemaking using the novel winemaking approach described herein. Further investigation would be required to assess strain effects on wine aroma, mouth feel and taste, however, this novel winemaking approach may offer significant industry efficiencies.

Down-regulation of photosynthesis following girdling, but contrasting effects on fruit set and retention, in two sweet cherry cultivars.

Sweet cherry (Prunus avium) trees were manipulated to analyse the contribution of soluble sugars to sink feedback down-regulation of leaf net CO2 assimilation rate (Anet) and fruit set and quality attributes. Total soluble sugar concentration and Anet were measured in the morning on fully expanded leaves of girdled branches in two sweet cherry cultivars, 'Kordia' and 'Sylvia' characterised typically by low and high crop load, respectively. Leaves on girdled trees had higher soluble sugar concentrations and reduced Anet than leaves on non-girdled trees. Moreover, RuBP carboxylation capacity of Rubisco (Vcmax) and triose-phosphate utilisation (TPU) were repressed in the girdled treatments, despite Jmax remaining unchanged; suggesting an impairment of photosynthetic capacity in response to the girdling treatment. Leaf Anet was negatively correlated to soluble sugars, suggesting a sink feedback regulatory control of photosynthesis. Although there were significantly less fruit set and retained in 'Kordia' than 'Sylvia'; girdling had contrasting effects in each cultivar. Girdling significantly increased fruit set and fruitlet retention in 'Sylvia' cultivar, but had no effect in 'Kordia' cultivar. We propose that low inherent sink demand for photoassimilates of 'Kordia' fruit could have contributed to the low fruit retention rate, since both non-girdled and girdled trees exhibited similar retention rate and that increases in foliar carbohydrates was observed above the girdle. In 'Sylvia' cultivar, the carbohydrate status may be a limiting factor for 'Sylvia' fruit, since girdling improved both fruit set and retention, and leaf soluble solids accumulation.

The effect of increased concentrations of homocysteine on the concentration of (E)-4-hydroxy-2-nonenal in the plasma and cerebrospinal fluid of patients with Alzheimer's disease.

There is evidence that increased blood concentrations of homocysteine may be a risk factor for Alzheimer's disease. (E)-4-hydroxy-2-nonenal (HNE) is a neurotoxic product of lipid peroxidation that is increased in the ventricular fluid and brains of patients with Alzheimer's disease. We measured the concentrations of homocysteine, HNE, vitamin B(12) and folate in the plasma of 27 patients with Alzheimer's disease and 25 control subjects. There was a statistically significant increase in the plasma concentration of homocysteine (P < 0.001) and HNE (P < 0.001) in the Alzheimer's disease patients compared to the control group. There was a significant decrease in the plasma concentration of vitamin B(12) (P < 0.001) and folate (P = 0.002) in the Alzheimer's group compared to the controls. There was a significant positive correlation between the plasma concentrations of homocysteine and HNE in the patients with Alzheimer's disease (r = 0.661, P < 0.001). A significant negative correlation was found between the plasma concentration of homocysteine and the plasma concentrations of vitamin B(12) (r = -0.605, P = 0.0006) and folate (r = 0.586, P = 0.001). We also measured the concentrations of homocysteine, HNE, vitamin B(12) and folate in the cerebrospinal fluid (CSF) of 8 patients with Alzheimer's disease compared to 6 control subjects. The concentrations of homocysteine (P = 0.032) and HNE (P = 0.001) were significantly higher in the CSF of Alzheimer's patients than in the control subjects. There were significant positive correlations between the CSF concentrations of homocysteine and HNE (r = 0.924, P = 0.001). There was also a significant positive correlation between the plasma concentration of homocysteine and the CSF concentrations of homocysteine (r = 0.850, P = 0.007) and HNE (r = 0.092, P = 0.002). These results demonstrate that there is a relationship between increased homocysteine concentrations and increased HNE concentrations in Alzheimer's disease.

Matrix metalloproteinase inhibitors in rheumatic diseases.

The rheumatic diseases continue to represent a significant healthcare burden in the 21st century. However, despite the best standard of care and recent therapeutic advances it is still not possible to consistently prevent the progressive joint destruction that leads to chronic disability. In rheumatoid arthritis and osteoarthritis this progressive cartilage and bone destruction is considered to be driven by an excess of the matrix metalloproteinase (MMP) enzymes. Consequently, a great number of potent small molecule MMP inhibitors have been examined. Several MMP inhibitors have entered clinical trials as a result of impressive data in animal models, although only one MMP inhibitor, Ro32-3555 (Trocade), a collagenase selective inhibitor, has been fully tested in the clinic, but it did not prevent progression of joint damage in patients with rheumatoid arthritis. The key stages and challenges associated with the development of an MMP inhibitor in the rheumatic diseases are presented below with particular reference to Trocade. It is concluded that the future success of MMP inhibitors necessitates a greater understanding of the joint destructive process and it is hoped that their development may be accompanied with clearer, more practical, outcome measures to test these drugs for, what remains, an unmet medical need.

Free radical formation in X-irradiated crystals of 2'-deoxycytidine hydrochloride. Electron magnetic resonance studies at 10 K.

Single crystals of deoxycytidine hydrochloride (CdR.HCl) have been X-irradiated at 10 K with doses up to about 150 kGy and studied using 24 GHz (K-band) EPR, ENDOR and FSE spectroscopy. In this system, the cytosine base is protonated at the N3 position. Nine different radicals were characterized and identified. Three of these are ascribed to three versions of the one-electron reduced species, probably differing in their protonation state. Radicals formed by net hydrogen addition to the cytosine C5 and C6 positions were observed at 10 K. The hydrogen-abstraction radical at the deoxyribose C1' position most probably results from initial oxidation of the base. The remaining radical species are all localized to the sugar moiety, representing products formed by net hydrogen abstraction from three of the five available carbons of the deoxyribose sugar. The lack of base-centered oxidation products as well as the structures of the one-electron reduced species is rationalized by considering the specific proton donor-acceptor properties of this crystalline lattice in comparison with similar systems.

Associations of demographic and health-related characteristics with prostate cancer screening in Washington State.

This report describes associations of demographic and health-related characteristics with use of prostate cancer screening. Data are from a random-digit dial survey of Washington State residents. Analyses are restricted to men ages 40-79 years (n = 332) and examine both digital rectal examination (DRE) and blood tests for prostate-specific antigen (PSA) in the previous 2 years. Results are adjusted to be representative of the state's population. In 1996, 53.6% of men received either DRE, PSA, or both. Among those screened, 42% received DRE alone, 15% PSA alone, and 43% both PSA and DRE, and the percentages of men receiving PSA increased markedly with age (30%, ages 40-49 years; 58%, ages 50-59 years; and 77%, ages 60-79 years). After control for other demographic characteristics, the relative odds for any prostate cancer screening were 5.5 for ages 60-79 versus 40-49 years, 2.4 for 16+ versus < or = 12 years of education, and 4.0 for 2+ versus no physician visits in the previous 2 years (all P < 0.05). Characteristics generally associated with good health, including regular exercise and low fat and high fruit and vegetable intakes, were also significantly associated with prostate cancer screening. In conclusion, in 1996, approximately one-half of the men in Washington State over age 40 years had received prostate cancer screening in the previous 2 years. Few men were screened with PSA alone, and the use of PSA as part of prostate cancer screening increased markedly with age. Because PSA screening increases detection of prostate cancer, epidemiological studies of health behavior and cancer risk must carefully control for screening history to avoid detection bias.

Electron paramagnetic resonance and electron nuclear double resonance studies of X-irradiated crystals of cytosine hydrochloride. Part I: free radical formation at 10 K after high radiation doses.

Anhydrous single crystals of cytosine hydrochloride (protonated at N3) have been X-irradiated at 10 K and studied using K-band EPR, ENDOR and FSE spectroscopy. At least seven radicals were present at 10 K after X irradiation with a dose of about 150 kGy. Two different protonation states of the one-electron reduced cytosine cation were observed: an amino-protonated species (R1) and the pristine one-electron reduced species (R2) with zero net charge. Apparently three deprotonated versions of the one-electron oxidized cytosine cation were formed: the amino-deprotonated cation (R3), an N3-deprotonated cation (R4) and an N1-deprotonated cation (R5). Finally, two products formed by net hydrogen addition to the cytosine base were observed: a C5 hydrogen-addition radical (R6) and a C6 hydrogen-addition radical (R7). The crystalline lattice of cytosine hydrochloride is characterized in part by a cytosine base initially protonated at the N3-position, thus forming a cytosine base cation, and in part by an extended network of hydrogen bonding involving the chlorine anions. Proton transfer properties of pristine one-electron oxidation and reduction base products in this lattice are discussed and are suggested as explanations of the unusual multitude of positions for deprotonation of the one-electron oxidized species as well as for the two protonation states of the reduction product observed. The magnetic parameters for the amino-protonated species R1 agree well with those extracted from previous studies of cytosine derivatives in frozen solutions and in various glasses.

Radiation damage to DNA base pairs. II. Paramagnetic resonance studies of 1-methyluracil x 9-ethyladenine complex crystals X-irradiated at 10 K.

Single crystals of the co-crystalline complex of 1-methyluracil and 9-ethyladenine were X-irradiated and studied using EPR, ENDOR and FSE spectroscopic techniques at 10 K. All together seven radicals were identified, and experimental evidence for at least one more species, as well as for a very low population of radical pairs, is available. Oxidation and reduction products appear to be stabilized at both base constituents of the pair. Of the 1-methyluracil moiety, the product formed by net hydrogen abstraction from the methyl group was observed, together with the 1-methyluracil anion and the 1-methyluracil-5-yl radical. From the 9-ethyladenine moiety, the N3-protonated 9-ethyladenine anion is stabilized. In addition, the 9-ethyladenine cation as well as traces of the amino-deprotonated cation were observed, together with the C8-H hydrogen adduct. The presence of oxidation and reduction products in each of the two bases may indicate that negligible energy transfer takes place between them. This behavior is different from that observed in the similar pair of 1-methylthymine-9-methyladenine. There also seems to be minor proton exchange between the two stacks of molecules: Interbase protonation-deprotonation channeled through the hydrogen-bonding scheme seems to be almost completely suppressed.

Electron spin resonance and electron nuclear double resonance study of X-irradiated deoxyadenosine: proton transfer behavior of primary ionic radicals.

A study of deoxyadenosine crystals (anhydrous form) after irradiation at 10 K found four base-centered radicals and one sugar-centered radical. Radical R1, thermally stable to about 100 K and photobleachable easily with white light, was the product of deprotonation at the amino group by the primary radical cation. Radical R2, also thermally stable to about 100 K, was the product of protonation at N3 of the primary radical anion. Radical R3, stable to about 170 K, was centered in the deoxyribose moiety and evidently was the result of net hydrogen abstraction from C4'. Radicals R4 and R5 were the C2 and C8 H-addition products with couplings typical of those species. Both R4 and R5 were formed at 10 K and were stable at room temperature. The behavior of R1 in several systems provides additional evidence for significant involvement of the hydrogen-bonding environment in controlling the stabilization (or formation) of radicals resulting directly from ionization, as described previously (Radiat. Res. 131, 272-284, 1992). From comparison of amino-group hydrogen-bonding environments in which radicals with the structure of R1 were stabilized, we conclude that oxygen atoms as proton acceptors are important in permitting the charge and spin separation necessary for radical stabilization. In particular, oxygens of ROH structures seem most efficient by readily permitting a multi-proton shuffle through a mechanism amounting to proton exchange. The collective results show that stabilization of these products is unlikely unless the charge and spin can be separated by at least one intervening molecule.

BPRS syndrome scales during the course of an episode of psychiatric illness.

This report examines changes in symptom levels on the four major syndrome scales from the Brief Psychiatric Rating Scale (BPRS): thought disturbance, paranoid disturbance, anxiety/depression, and emotional withdrawal/motor retardation. Baseline BPRS ratings were obtained during the first week of hospitalization for an acute episode of psychiatric illness, in 120 patients with schizophrenia, schizoaffective disorder, and depression. BPRS ratings were carried out in the week prior to discharge. Findings indicated that patients with schizoaffective disorder showed a greater magnitude of general clinical improvement than schizophrenics, although both groups had comparable improvement on thought disorder from admission to discharge. Paranoid symptoms did not recover as completely among schizophrenics compared to schizoaffective disorder patients. As expected, anxiety and depression symptoms remitted most prominently among the depressed inpatients.

Where are the sugar radicals in irradiated DNA?

Numerous sugar radicals have been observed by EPR and ENDOR spectroscopy in X-irradiated nucleosides and nucleotides. However, no sugar radicals have been observed in irradiated DNA. One possibility exists that sugar radicals may be present in relatively small abundance and therefore have so far escaped detection. Another possibility is that each of the five carbon-centered H-abstraction sugar radicals may exist in a wide range of conformations. Adding together various groups of radicals with different hyperfine couplings and anisotropic g factors may result in very broad EPR lines which would be difficult to detect. Using the crystal structure of a B-DNA dodecamer, the conformations of the five H-abstraction sugar radicals have been computed at all sites. The X-band EPR spectra for each radical were then simulated using typical alpha-proton and beta-proton couplings computed from the various radical conformations. The results suggest considerable broadenings of the EPR lines for the C2', C3' H-abstraction radicals. Composite EPR spectra were simulated by adding 15% of an H-abstraction radical to the DNA spectrum. The results indicate that the outer lines of the C1' radical are visible and should be easy to identify. The broad spectra of the C2' and C3' radicals are barely visible. The simulated spectra of the C4' and C5' radicals are basically doublets and are obscured by the DNA signal.

Neuropsychological dysfunction and clinical outcome in psychiatric disorders: a two-year follow-up study.

The presence of neuropsychological disturbances in schizophrenia and mood disorders raises the question that cognitive impairments might contribute to poor outcome. This report examines changes in neuropsychological performance from hospitalization to a 2-year follow-up evaluation in relation to psychosocial outcome. Findings indicated that unfavorable clinical outcome is associated with marginal changes in neuropsychological performance, whereas good outcome status is associated with neuropsychological improvement. Neuropsychological improvement may thus require a stable period of favorable psychosocial recovery, in schizophrenia and schizoaffective disorder, as well as major mood disorder syndromes.

Radiation damage to DNA base pairs. I. Electron paramagnetic resonance and electron nuclear double resonance study of single crystals of the complex 1-methylthymine.9-methyladenine X-irradiated at 10K.

Single crystals of the complex 1-methylthymine.9-methyl-adenine were X -irradiated at 10 and at 65 K and studied in the temperature range 10 to 290 K using K-band EPR, ENDOR and field-swept ENDOR (FSE) techniques. The EPR and ENDOR spectra are dominated by two major and four minor resonances. The two major resonances are: MTMA1, the well-known radical formed by net hydrogen abstraction fr om the CS methyl group of the thymine moiety, and MTMA2, the radical formed by net hydrogen abstraction from the N1 methyl group of the thymine moiety. The latter product has not been observed previously in any 1-methylthymine derivative. The four minor resonances are: MTMA3, the anion of 1-methylthymine, possibly protonated at the O4 position; MTMA4, the well-known species formed by net hydrogen addition to C6 of the thymine moiety; MTMA5, the species formed by net hydrogen addition to C2 of the adenine moiety; and MTMA6, the species formed by net hydrogen addition to C8 of the adenine moiety. Radical MTMA3, the O4-protonated thymine anion, was clearly detected at 10 K, but upon thermal annealing at 40 K the lines began to disappear. In crystals irradiated at 65 K MTMA3 was only weakly present. Radical MTMA2 decayed at about 250 K with no detectable successor, and radical MTMA5 disappeared at about 180 K. It was not possible to learn from the d ata if MTMA5 transformed into MTMA6. The radical distribution in the 1-methylthymine.9-methyladenine crystal system is different from that in crystals of the individual components. Reasons for this behavior are discussed in light of the hydrogen bonding schemes and molecular stacking interactions in each of the crystals. An important feature is the concept of excited-state transfer from the adenine to the thymine moiety, followed by dehydrogenation at the thymine Nl-methyl group, the mechanism resulting in radical MTMA2.

Free radical formation in single crystals of 9-methyladenine X-irradiated at 10 K. An electron paramagnetic resonance and electron nuclear double resonance study.

Single crystals of 9-methyladenine were X-irradiated at 10 K and at 65 K and were studied using K-band EPR, ENDOR and field-swept ENDOR (FSE) techniques in the temperature range 10 K to 290 K. Three major radicals are stabilized in 9-methyladenine at 10 K. These are: MA1, the adenine anion, probably protonated at N3; MA2, the species formed by net hydrogen abstraction from the 9-methyl group; and MA3, the radical formed by net hydrogen addition to C8 of the adenine moiety. Radical MA1 decayed at about 80 K, possibly into the C2 H adduct (MA4). The other two species (MA2, MA3) were stable at room temperature. A fifth radical species was clearly present in the EPR spectra at 10 K but was not detectable by ENDOR. This species, which decayed above 200 K (possibly into MA3), remains unidentified. The radical population at room temperature is as described by previous authors. The mechanisms for radical formation in 9-methyladenine are discussed in light of the hydrogen bonding scheme and molecular stacking interactions.

ESR and ENDOR study of single crystals of deoxyadenosine monohydrate X-irradiated at 10 K.

Five free radicals have been detected by detailed ESR/ENDOR experiments on single crystals of deoxyadenosine monohydrate (AdRm), X-irradiated and observed at 10 K. In a previous study of adenosine (Radiat. Res. 117, 367-378, 1989), only the anion (protonated at N3) and the cation (deprotonated at the exocyclic NH2) were detected at 10 K. In AdRm, Radical R1 is the N3-protonated anion, similar to that observed previously in adenosine. Radical R3 is a C5' hydroxyalkyl radical formed by net H-abstraction from C5'. A second sugar radical is formed by net C1' H-abstraction. Two other base radicals observed in AdRm at 10 K are the C2 and C8 H-addition radicals. The C2 H-addition radical (Radical R5) exhibits inequivalent methylene hydrogen couplings of 5.43 and 3.29 mT, while in the C8 H-addition radical (Radical R6) the couplings are somewhat more equivalent (3.63 and 4.17 mT). No link between RAdical R1 and the H-addition radicals has been observed. The reduced base appears to protonate rapidly even at 10 K, while at the same time both H-addition radicals are clearly present. On warming, Radical R1 appears to decay at about 80 K with no apparent successor. Although no base cation was stabilized in AdRm at 10 K, it is interesting to note that Radicals R3 and R4 could both be formed as the result of deprotonation of primary oxidation products.

Phase II trial of piroxantrone in metastatic gastric adenocarcinoma.

Piroxantrone, a synthetic intercalating agent, was studied in patients with advanced, measurable gastric adenocarcinoma who had not received prior chemotherapy. The starting piroxantrone dose was 150 mg/m2 given intravenously over 1 hour on day 1 and repeated every 21 days. Response and toxicity could be evaluated in 15 patients. No complete, partial, or minor responses were observed. Toxic effects included granulocytopenia, anemia, vomiting, nausea, anorexia, fatigue, stomatitis, alopecia, hyperbilirubinemia, and increased alkaline phosphatase levels. At the stated dose and schedule, piroxantrone does not possess significant activity against advanced gastric cancer.

Radical ions and their reactions in DNA constituents: ESR/ENDOR studies of radiation damage in the solid state.

This updating review contains the results of recent ESR/ENDOR experiments on DNA constituents in the solid state. The compounds reviewed include single crystals of the DNA bases, nucleosides, and nucleotides. The emphasis is on low-temperature ENDOR results. Typical experiments involve irradiations at or near helium temperatures in attempts to determine the primary radiation-induced oxidation and reduction products. The use of the ENDOR technique allows one to determine the protonation state of the initial products. Subsequent warming of the sample facilitates a study of the reactions that these primary products undergo. A critical review of previous ESR experiments is considered to see if these results conflict with the newer ENDOR results. Finally, a summary is provided to show which of the numerous results discussed may have some relevance to understanding the radiation chemistry of DNA.