Cognitive Functioning in Survivors of Hematopoietic Stem Cell Transplantation Compared With a Matched General Population Sample-The Maastricht Observational Study of Late Effects After Stem Cell trAnsplantation Study.

Although cognitive problems can recover over time, a subgroup of hematopoietic stem cell transplantation (HCT) survivors experience persistent cognitive problems in the long term. Despite these implications, studies assessing cognitive functioning in HCT survivors are limited. The aim of the present study was (1) to quantify the prevalence of cognitive impairment in patients treated with HCT who survived at least 2 years and to compare these with a matched reference group representing the general population; (2) to identify potential determinants of cognitive functioning within the HCT survivor group. Within the single-center Maastricht Observational study of late effects after Stem cell trAnsplantation, cognitive performance was assessed by a neuropsychological test battery divided into 3 cognitive domains: memory, information processing speed, and executive function and attention. An overall cognition score was calculated as the average of the domain scores. A total of 115 HCT survivors were group-matched on a 1:4 ratio to the reference group by age, sex, and level of education. Regression analyses adjusted for different sets of covariates including demographic and health- and lifestyle-related factors were used to test for differences in cognition between HCT survivors and the reference group resembling the general population. A limited set of clinical characteristics (diagnosis, type of transplant, time since treatment, conditioning regimen with total body irradiation and age at time of transplantation) were assessed as potential determinants of neurocognitive dysfunction among HCT survivors. Cognitive impairment was defined as scores in the cognitive domains < -1.5 standard deviation (SD) from what can be expected based on someone's age, sex, and education. The mean age at time of transplantation was 50.2 (SD ± 11.2) years, and the mean number of years after transplant was 8.7 (SD ± 5.7) years. The majority of HCT survivors were treated with autologous HCT (n = 73 [64%]). The prevalence of cognitive dysfunction was 34.8% in HCT survivors and 21.3% in the reference group (p = .002.) When adjusted for age, sex, and level of education, HCT survivors had a worse overall cognition score (b = -0.35; 95% confidence interval [CI], -0.55 to -0.16; p < .001), translating into 9.0 years of higher cognitive age. Analyses of specific cognitive domain scores showed that HCT survivors scored worse on memory (b = -0.43; 95% CI, -0.73 to -0.13; p = .005), information processing speed (b = -0.33; 95% CI, -0.55 to -0.11; p = .003), and executive function and attention (b = -0.29; 95% CI, -.55 to -.03; p = .031) than the reference group. The odds of cognitive impairment were on average 2.4 times higher among HCT survivors than the reference group (odd ratio = 2.44; 95% CI, 1.47-4.07; p = .001). Within the HCT survivor group none of the tested clinical determinants of cognitive impairment were significantly associated with cognition. This cohort study showed evidence for worse cognitive functioning in HCT survivors encompassing all three cognitive domains, respectively memory, information processing speed, and executive and attention compared to a reference group that represents the general population translating into nine years of faster cognitive ageing in HCT survivors than can be expected based on their chronological age. It is important to increase awareness for signs of neurocognitive dysfunction after HCT in clinicians and HCT survivors.

Comorbidity clusters in patients with moderate-to-severe OSA.

PURPOSE: Obstructive sleep apnea (OSA) is a prevalent and multifaceted disease. To date, the presence and severity of objectively identified comorbidities and their association with specific OSA phenotypes, CPAP adherence, and survival remain to be elucidated. The aim of this study is to cluster patients with OSA based on 10 clinically important objectively identified comorbidities, and to characterize the comorbidity clusters in terms of clinical and polysomnographic characteristics, CPAP adherence, and survival. STUDY DESIGN AND METHODS: Seven hundred ten consecutive patients starting CPAP for moderate-to-severe OSA were included. Comorbidities were based on generally accepted cutoffs identified in the peer-reviewed literature. Self-organizing maps were used to order patients based on presence and severity of their comorbidities and to generate clusters. RESULTS: The majority of patients were men (80%). They were generally middle-aged (52 years) and obese (BMI: 31.5 kg/m2). Mean apnea-hypopnea index (AHI) was 41 ± 20 per h of sleep. More than 94% of the patients had one or more comorbidities with arterial hypertension, dyslipidemia, and obesity being the most prevalent. Nine comorbidity clusters were identified. The clinical relevance of these comorbidity clusters was highlighted by the difference in symptoms, PSG parameters, and cardiovascular risk. Also, differences in CPAP adherence, improvements in ESS, and long-term survival were present between the clusters. CONCLUSION: Comorbidity prevalence in patients with OSA is high, and different comorbidity clusters, demonstrating differences in cardiovascular risk, CPAP adherence, and survival, can be identified. These results further substantiate the need for a comprehensive assessment of patients with OSA beyond the AHI.

Low level detection of Benzene in Food Grade Hexane by Ultraviolet Spectrophotometry.

A simple, sensitive, and accurate Ultraviolet Spectrophotometric method has been developed and validated for the determination of Benzene in Food Grade Hexane. Benzene in spectroscopic grade Hexane shows vibrational fine structure having four well resolved peaks. In the wavelength range 240-270nm, peak at 255nm is considered for the method development. Beer's law was obeyed in the concentration range of 0.6-10.0µLL(-1), with correlation coefficient, 0.9999, detection limit 0.2µLL(-1) and quantitation limit 0.6µLL(-1) are established. Percentage recovery studies showed that the method was not affected by the presence of other solvents having the similar boiling range with Hexane. The method was validated by determining its accuracy and precision which proves suitability of the developed method for the routine determination of Benzene in Food Grade Hexane. The proposed method has been applied successfully for the analysis of the Food Grade Hexane.

Integrity of N- and C-termini is important for E. coli Hsp31 chaperone activity.

Hsp31 is a stress-inducible molecular chaperone involved in the management of protein misfolding at high temperatures and in the development of acid resistance in starved E. coli. Each subunit of the Hsp31 homodimer consists of two structural domains connected by a flexible linker that sits atop a continuous tract of nonpolar residues adjacent to a hydrophobic bowl defined by the dimerization interface. Previously, we proposed that while the bowl serves as a binding site for partially folded species at physiological temperatures, chaperone function under heat shock conditions requires that folding intermediates further anneal to high-affinity binding sites that become uncovered upon thermally induced motion of the linker. In support of a mechanism requiring that client proteins first bind to the bowl, we show here that fusion of a 20-residue-long hexahistidine tag to the N-termini of Hsp31 abolishes chaperone activity at all temperatures by inducing reversible structural changes that interfere with substrate binding. We further demonstrate that extending the C-termini of Hsp31 with short His tags selectively suppresses chaperone function at high temperatures by interfering with linker movement. The structural and functional sensitivity of Hsp31 to lengthening is consistent with the high degree of conservation of class I Hsp31 orthologs and will serve as a cautionary tale on the implications of affinity tagging.

A genetic approach for controlling the binding and orientation of proteins on nanoparticles.

Although silver nanoparticles are excellent surface enhancers for Raman spectroscopy, their use to probe the conformation of large proteins at interfaces has been complicated by the fact that many polypeptides adsorb weakly or with a random orientation to colloidal silver. To address these limitations, we sought to increase binding affinity and control protein orientation by fusing a silver-binding dodecapeptide termed Ag4 to the C-terminus of maltose-binding protein (MBP), a well-characterized model protein with little intrinsic silver binding affinity. Quartz crystal microbalance measurements conducted with the MBP-Ag4 fusion protein revealed that its affinity for silver (Kd approximately 180 nM) was at least 1 order of magnitude higher than a control protein, MBP2, containing a non-silver-specific C-terminal extension. Under our experimental conditions, MBP-Ag4 SERS spectra exhibited 2-4 fold higher signal-to-background relative to MPB2 and contained a number of amino acid-assigned vibrational modes that were either weak or absent in control experiments performed with MBP2. Changes in amino acid-assigned peaks before and after MBP-Ag4 bound maltose were used to assess protein orientation on the surface of silver nanoparticles. The genetic route described here may prove useful to study the orientation of other proteins on a variety of SERS-active surfaces, to improve biosensors performance, and to control functional nanobiomaterials assembly.

Ammonium dithionate- a new material for highly sensitive EPR dosimetry.

Polycrystalline ammonium dithionate has been examined for its radiation response in the low dose range (<5Gy) using EPR technique. The SO(3)(-) radical ion was detected as a single EPR line with a peak-to-peak derivative width of ca. 0.44mT in irradiated samples and its intensity was found to vary linearly with dose. At equal and moderate settings of microwave power and modulation amplitude ammonium dithionate was at least seven times more sensitive than l-alanine which is the most common EPR dosimeter standard. Pulse experiments were performed on the powder samples to obtain the longitudinal relaxation time. These and microwave saturation experiments served to indicate the optimal microwave power to be applied during measurements as an EPR dosimeter for best sensitivity of this material. It is thus claimed that ammonium dithionate has excellent potential to become an EPR dosimeter with a low limit of the measurable dose for cases where tissue equivalence is not required or can be corrected for.

Conformational control of inorganic adhesion in a designer protein engineered for cuprous oxide binding.

Combinatorial selection of peptides that bind technological materials has emerged as a valuable tool for room-temperature nucleation and assembly of complex nanostructured materials. At present, the parameters that control peptide-solid binding are poorly understood, but such knowledge is needed to build the next generation of hybrid materials. Here, we use a derivative of the DNA binding protein TraI engineered with a disulfide-bonded cuprous oxide binding sequence called CN225 to probe the influence of sequence composition and conformation on Cu2O binding affinity. We previously reported a statistically significant enrichment in paired arginines (RR) among a family of cuprous oxide binding peptides and hypothesized that this is a key motif for binding. However, systematic alanine (A) substitutions in the CN225 RR motif (creating RA, AR, and AA pairs) do not support the hypothesis that RR is critical for Cu2O binding by CN225. Instead, we find that the presentation of the peptide in a disulfide-constrained loop (i.e., the conformation present during combinatorial selection) is crucial for binding to the metal oxide. Our results suggest that caution should be exerted when extrapolating from statistical data and that, in some cases, conformation is more important than composition in determining peptide-inorganic adhesion.

Development of nickel-doped lithium formate as potential EPR dosimeter for low dose determination.

EPR dosimetry employing L-alpha-alanine has been in vogue during the past few years, due to its tissue equivalence and linear dose response. However, L-alpha-alanine dosimetry has been improved during the past years, the sensitivity of this material is still too low for clinical applications. Polycrystalline lithium formate doped with NiCl2 was therefore examined for radiation response in the dose range of clinical interest (<5 Gy) using CW EPR and pulse EPR techniques. At equal and moderate settings of microwave power and modulation amplitude lithium formate doped with 1.6 wt% of NiCl2 was almost four times more sensitive compared to L-alpha-alanine, which is the most common EPR dosimeter standard. It was shown that the nickel-doped lithium formate has an excellent radiation response with a low limit of the measurable dose, and a linear dose response in the range 1-5 Gy. The relaxation and power saturation studies showed that high microwave power can be applied during measurements to improve the sensitivity of this material as an EPR dosimeter. These results show that lithium formate doped with Ni(II) exhibits promising properties required for further development of an EPR dosimeter in the dose range typical for clinical dosimetry.

Dynamical effects and ergodicity in the dipolar glass phase: evidence from time-domain EPR and phase memory time studies of AsO(4)(4-) in Rb(1-x)(NH(4))(x)H(2)PO(4) (x = 0,0.5,1).

Three-pulse electron spin echo envelope modulation (ESEEM), hyperfine sublevel correlation spectroscopy (HYSCORE) investigations and two-pulse electron spin echo (ESE) measurements of phase memory time T(M), were carried out, in the 20-200 K temperature range, on an AsO(4)(4-) paramagnetic probe stabilized in RbH(2)PO(4) (RDP), NH(4)H(2)PO(4) (ADP), and dipolar glass Rb(0.5)(NH(4))(0.5)H(2)PO(4) (RADP). The results obtained on ADP revealed hyperfine interaction of the probe ion with the (14)N of the ammonium ion, the coupling constant satisfying the condition of 'cancellation' at a field of 480 mT. The ammonium ion was found to be in two different sites in ADP, which became indistinguishable on the formation of dipolar glass RADP. These results were confirmed by HYSCORE spectral measurements. The fast Fourier transform (FFT) spectra of three-pulse ESEEM decays have clearly revealed the interaction with protons in the [Formula: see text] bond both in ADP and RDP; and in RADP with an averaged coupling constant. The phase memory times in RADP exhibited strong temperature dependence and were found to be dependent on the nuclear spin quantum number m(I) of (75)As. The temperature dependence of T(M) exhibited a well-defined maximum around 90 K, coinciding with the temperature of onset of 'freezing' in Rb(0.5)(NH(4))(0.5)H(2)PO(4). This is symptomatic of dynamic fluctuations in the dipolar glass phase, with onset around 150 K, going through a maximum around 90 K and slowing down on further cooling. These results suggest that in RADP, a dynamical mechanism with progressive slowing down below 90 K is operative in the glass formation. This implies that the RADP system, with x = 0.5, exists in an ergodic relaxor (R)-state in the 20-200 K temperature range wherein every fluctuating monodomain can be viewed as statistically representative of the whole sample.

Enhanced sensitivity of lithium dithionates doped with rhodium and nickel for EPR dosimetry.

Electron paramagnetic resonance (EPR) studies of X-irradiated lithium dithionate, Li(2)S(2)O(6) x 2H(2)O, doped with Ni and Rh have shown that these impurities enhance the yield of radicals formed by X-irradiation at room temperature. The signal in the doped samples, measured peak-to-peak of the single EPR derivative line attributed to the SO(3)(-) anion was about 3-4 times that of the pure lithium dithionate and more than 10 times stronger than the alanine signal. These impurities also shortened the spin-lattice relaxation time, T(1), which gives the possibility to measure the doped samples at a higher microwave power. This implies that sensitivity could be further enhanced in the already sensitive EPR dosimeter material lithium dithionate.

Plasma leptin concentration increases early during highly active antiretroviral therapy for acquired immunodeficiency syndrome, independent of body weight.

Leptin, the protein product of the obese gene (ob), is secreted by adipocytes. Circulating leptin levels correlate with fat mass in humans, including individuals infected with HIV. Leptin serves as an adipostatic hormone, a permissive factor for reproduction and a modulator of immune function. Leptin is a cytokine, and has been demonstrated to enhance CD4 cell proliferation and IL-2 secretion from CD4 cells in vitro. The role of leptin in HIV-positive patients treated with highly active antiretroviral therapy (HAART) has not been well defined. We haveevaluated leptin levels in HIV-infected individualsduringthe early phase of HAART. We measured plasma leptin levels in 15 antiretroviral-naive HIV positive patients at baseline and after 1 and 4 weeks of HAART. After the first week of therapy, mean leptin level and CD4 count were increased compared to baseline, 6.0 vs 7.2 ng/ml (p = 0.004) and 377 vs 432 cells/ul (p = 0.014), respectively. In contrast, mean body mass index (BMI) remained unchanged 27.0 vs 26.8 kg/m2 (p < 0.08). After four weeks of therapy, leptin and BMI values were unchanged compared to baseline, 6.0 vs 5.9 (p < 0.4) and 27.0 vs 26.9 (p < 0.5), respectively, whereas CD4 count continued to increase to 491 cells/ul (p < 0.012 compared to baseline). These data demonstrate an early transient increase in plasma leptin levels in HIV positive patients initiated on HAART, despite a lack of change in BMI. It is unclear if the transient increase in leptin is related to its role as a cytokine, a metabolic regulator, or reproductive factor.

Formates and dithionates: sensitive EPR-dosimeter materials for radiation therapy.

Polycrystalline formates and dithionates are promising materials for EPR dosimetry, as large yields of radiation induced stable radicals are formed with a linear dose response. Rapid spin relaxation rates were detected in many of the substances, indicating that a high microwave power can be applied during EPR acquisition in order to improve sensitivity. Different techniques used to further improve the sensitivity, such as the replacement of 7Li with 6Li or exchange of protons with deuterons in the corresponding crystalline matrices and metal ion doping are discussed. It is concluded that formates and dithionates may be up to 10 times as sensitive as L-alpha-alanine.

51V(n, beta)52Cr reaction for neutron dosimetry: development and assessment of a spectrophotometric method for determination of Cr in vanadium at sub ppm level.

With a view to monitoring the changes in coloration caused by the nuclear reaction 51V(n, beta)52Cr in solution of vanadyl sulphate and using it for neutron dosimetry, electronic absorption spectra of vanadyl sulphate solutions were investigated at different concentrations of chromate impurity in micromolar range. It was observed that the presence of chromate enhances the absorptivity over a wide wavelength range serving essentially as a colouring agent for vanadium matrix, presumably due to charge transfer process. The absorbance at 380 nm varied linearly over a wide concentration range. The limit of detection of chromate obtained is shown to be adequate for detecting neutron-induced chemical transmutation of vanadium to chromium under standard reactor conditions, when used with long path length cells. It was observed that the absorbance does not change on electron irradiation, suggesting that radiolytic effects due to beta decay, if any, do not interfere in the measurement of neutron-induced changes. In addition to its potential for neutron dosimetry, this is the first report of a simple and direct method of estimation of Cr in vanadium matrix at sub ppm level.

Identification and characterization of Cu(2)O- and ZnO-binding polypeptides by Escherichia coli cell surface display: toward an understanding of metal oxide binding.

We have used the FliTrx cell surface display system to identify disulfide-constrained dodecapeptides binding to the semiconducting metal oxides Cu(2)O and ZnO. Sequence analysis of the inserts revealed that the two populations exhibit similar, yet subtly different patterns of amino acid usage. Both sets of binders were enriched in arginine, tryptophan, and glycine with a higher degree of positional preference in the case of Cu(2)O binders. Tyrosine, proline, and serine were underrepresented in both populations. Peptides binding electrodeposited Cu(2)O or ZnO with high avidity could be subdivided into two classes based on pI and hydrophilicity. In the hydrophilic and positively charged Class I binders, the Arg-X-X-Arg tetrapeptide appears to be implicated in metal oxide binding, whereas Arg-Arg and Arg-Lys pairs allow for discrimination between Cu(2)O and ZnO. Molecular dynamics simulations of the disulfide-constrained peptides suggest that the aforementioned motifs are important to properly orient two basic residues that are likely to contact the metal oxides. The implications of our results in understanding the rules governing the interaction between peptides and inorganic compounds and in their use for the design of hybrid nanoarchitectures are discussed.

The linker-loop region of Escherichia coli chaperone Hsp31 functions as a gate that modulates high-affinity substrate binding at elevated temperatures.

Precise control of substrate binding and release is essential for molecular chaperones to exert their protective function in times of stress. The mechanisms used are diverse and have been difficult to unravel. Escherichia coli heat-shock protein 31 (Hsp31) is a recent addition to the known complement of eubacterial chaperones. Crystallographic studies have revealed the presence of a hydrophobic bowl at the Hsp31 dimer interface and shown that the linker region connecting the two structural domains within each subunit is disordered. Together with a neighboring flexible loop, the linker caps a hydrophobic area adjacent to the bowl. Using a collection of Hsp31 mutants, we show that although both bowl and linker-loop-shielded residues participate in substrate binding, the latter are critical for protein capture at high temperature. Linker immobilization via an artificial disulfide bridge abolishes chaperone activity at elevated temperatures by precluding exposure of the underlying hydrophobic domain. We conclude that Hsp31 uses its linker-loop region as a thermally activated gate to control nonnative protein annealing to a high-affinity substrate-binding site. This simple yet efficient strategy to capture partially folded proteins under heat-shock conditions may be shared by other folding modulators.

Compounds of 6Li and natural Li for EPR dosimetry in photon/neutron mixed radiation fields.

Formates and dithionates of 6Li, enriched and 7Li in natural composition of Li offer a possibility to measure the absorbed dose from photons and thermal neutrons in a mixed radiation field for instance at a boron neutron capture therapy (BNCT) facility. Tests with formates and dithionates of enriched 6Li and lithium compounds with natural composition have been performed at the BNCT facility at Studsvik, Sweden. Irradiations have been performed at 3 cm depth in a Perspex phantom in a fluence rate of thermal neutrons 1.8 x 10(9) n cm(-2) s(-1). The compounds were also irradiated in a pure X-ray field from a 4MV linear accelerator at 5 cm depth in a phantom with accurately determined absorbed doses. The signal intensity and shape was investigated within 3 h after the irradiation. A single line spectrum attributed to the CO2- radical was observed after irradiation of lithium formate. An increase in line width occurring after neutron irradiation in comparison with photon irradiation of the 6Li sample was attributed to dipolar broadening between CO2- radicals trapped in the tracks of the alpha particles. A spectrum due to the SO3- radical anion was observed after irradiation of lithium dithionate. The signal amplitude increased using the 6Li in place of the Li with natural composition of isotopes, in studies with low energy X-ray irradiation. Due to the decreased line width, caused by the difference in g(N) and I between the isotopes, the sensitivity with 6Li dithionate may be enhanced by an order of magnitude compared to alanine dosimetry. After comprehensive examination of the different combinations of compounds with different amounts of 6Li and 7Li regarding dosimetry, radiation chemistry and EPR properties these dosimeter material might be used for dose determinations at BNCT treatments and for biomedical experiments. Interesting properties of the radical formation might be visible due to the large difference in ionization density of neutrons compared to photons.

Air quality status at selected locations in Hyderabad City.

A case study for assessing the air quality status is elaborated for Hyderabad city. Monitoring was carried out at 11 locations during March 2003. These observations on air quality status and AQEI predicts that most of the localities in Hyderabad are experiencing the air pollution stress and the trend is likely to worsen in near future if proper control measures are not implemented.

Physical behavior of purified and crude wax obtained from sunflower (Helianthus annuus) seed oil refineries and seed hulls.

The sunflower seed waxes obtained from two sources (i) seed hull as a standard and (ii) crude wax from oil refineries were studied for their crystallization, melting characteristics and morphology of crystals. The results of differential scanning calorimetry of wax obtained from seed hulls showed the melting temperature range of 13.18 degrees C with the onset at 62.32 degrees C, for purified wax, compared to the melting range of 24.73 degrees C with the onset at 42.3 degrees C. for crude wax. The enthalpy of fusion for both waxes were 57.55 mcal/mg and 7.63 mcal/mg, respectively. The DSC melt crystallization temperature range was 15.79 degrees C with the onset of 64.58 degrees C for purified wax and temperature range of 31.45 degrees C with an onset of 57.76 degrees C for crude wax. A similar pattern was observed of wax obtained from the crude wax of oil refineries. The enthalpy of crystallization was -64.27 mcal/mg and -7.67 mcal/mg, respectively. The purified wax obtained from the two sources (i) and (ii) were comparable with completion temperatures of 75.5 degrees C and 75.1 degrees C, respectively. The effect of inhibitor (lecithin) on crystallization of purified wax under light microscope and surface structure by scanning electron microscope were observed. Lecithin at 0.2% inhibited the crystallization but nucleation was unaltered. The wax crystal was inhibited to around 60% of the original size with 0.2% lecithin. It is concluded that the sunflower waxes studied were not comparable in their crystal properties of crude and purified states. Lecithin inhibited the crystallization of sunflower seed wax.

Hsp31, the Escherichia coli yedU gene product, is a molecular chaperone whose activity is inhibited by ATP at high temperatures.

The Escherichia coli chromosome contains several uncharacterized heat-inducible loci that may encode novel molecular chaperones or proteases. Here we show that the 31-kDa product of the yedU gene is an efficient homodimeric molecular chaperone that is conserved in a number of pathogenic eubacteria and fungi. Heat shock protein (Hsp) 31 relies on temperature-driven conformational changes to expose structured hydrophobic domains that are likely responsible for substrate binding. Complementing the function of refolding, remodeling, and holding chaperones, Hsp 31 preferentially interacts with early unfolding intermediates and rapidly releases them in an active form after transfer to low temperatures. Although Hsp 31 does not appear to exhibit intrinsic ATPase activity, binding of ATP at high temperatures restricts the size or availability of the substrate binding site, thereby modulating chaperone activity. The possible role of ATP in coordinating the function of the cellular complement of molecular chaperones is discussed.