Infrequent Translation of a Nonsense Codon Is Sufficient to Decrease mRNA Level

In many organisms nonsense mutations decrease the level of mRNA. In the case of mammalian cells, it is still controversial whether translation is required for this nonsense-mediated RNA decrease (NMD). Although previous analyzes have shown that conditions that impede translation termination at nonsense codons also prevent NMD, the residual level of termination was unknown in these experiments. Moreover, the conditions used to impede termination might also have interfered with NMD in other ways. Because of these uncertainties, we have tested the effects of limiting translation of a nonsense codon in a different way, using two mutations in the immunoglobulin μ heavy chain gene. For this purpose we exploited an exceptional nonsense mutation at codon 3, which efficiently terminates translation but nonetheless maintains a high level of μ mRNA. We have shown 1) that translation of Ter462 in the double mutant occurs at only ∼4% the normal frequency, and 2) that Ter462 in cis with Ter3 can induce NMD. That is, translation of Ter462 at this low (4%) frequency is sufficient to induce NMD.

Concentrations and distribution of organochlorine pesticides in shellfish from Changjiang estuary

To analyze and evaluate the status of organochlorine pollutants in the Changjiang (Yangtze River) estuary and adjacent waters, the concentrations of hexachlorocyclohexane (HCHs) and dichlorodiphenyltrichloroethane (DDTs) in shellfish collected in study area from 2006 to 2007 were determined with gas chromatography (GC). The concentration range of HCHs was (ND-12.13)x10(-3) mg/kg wet weight and averaged at 0.54x10(-3) mg/kg while the concentration of DDTs was in the range of (4.06-281.73) x10(-3) mg/kg with a mean of 57.52x10(-3) mg/kg in the survey areas. The concentrations of DDTs in the shellfish were higher than HCHs', so that DDTs could be considered as typical organochlorine pollutants in the areas. The concentrations of DDTs in the shellfish were higher than HCHs', so that DDTs could be considered as typical organochlorines pollutants. The HCHs in all the shellfish conformed to the first level of criterion (0.02 mg/kg) of the Marion Biology Quality (GB 18421-2001), and that of DDTs in most samples were beyond the first level (0.01 mg/kg) but conformed to the second level (0.10 mg/kg). On average, alpha-HCH and delta-HCH occupied the most part of HCHs, while O,P'-DDT and P,P'-DDT occupied the most part of DDTs. The concentrations of organocholorine pesticides in shellfish samples varied in site and in species. The highest level occurred at the Shengsi (SS), followed by Yangkougang (YKG), Lvsi (LS), Dongyuan (DY) and Beibayao (BBY), low concentrations were observed at Changsha (CS), Beidaodi (BDD), and Gouqi (GQ). The concentration of HCHs and DDTs in most sites decreased clearly from 2006 to 2007 except for YKG, DY, BDD, LYS, and SS. All of above results suggested that the study area was slightly affected by organochlorine pesticide, special by DDTs.

The effect of key process operational conditions on enhanced biological phosphorus removal from wastewater

Enhanced biological phosphorus removal (EBPR) is the most economic and sustainable option used in wastewater treatment plants (WWTPs) for phosphorus removal. In this process it is important to control the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), since EBPR deterioration or failure can be related with the proliferation of GAOs over PAOs. This thesis is focused on the effect of operational conditions (volatile fatty acid (VFA) composition, dissolved oxygen (DO) concentration and organic carbon loading) on PAO and GAO metabolism. The knowledge about the effect of these operational conditions on EBPR metabolism is very important, since they represent key factors that impact WWTPs performance and sustainability. Substrate competition between the anaerobic uptake of acetate and propionate (the main VFAs present in WWTPs) was shown in this work to be a relevant factor affecting PAO metabolism, and a metabolic model was developed that successfully describes this effect. Interestingly, the aerobic metabolism of PAOs was not affected by different VFA compositions, since the aerobic kinetic parameters for phosphorus uptake, polyhydroxyalkanoates (PHAs) degradation and glycogen production were relatively independent of acetate or propionate concentration. This is very relevant for WWTPs, since it will simplify the calibration procedure for metabolic models, facilitating their use for full-scale systems. The DO concentration and aerobic hydraulic retention time (HRT) affected the PAO-GAO competition, where low DO levels or lower aerobic HRT was more favourable for PAOs than GAOs. Indeed, the oxygen affinity coefficient was significantly higher for GAOs than PAOs, showing that PAOs were far superior at scavenging for the often limited oxygen levels in WWTPs. The operation of WWTPs with low aeration is of high importance for full-scale systems, since it decreases the energetic costs and can potentially improve WWTP sustainability. Extended periods of low organic carbon load, which are the most common conditions that exist in full-scale WWTPs, also had an impact on PAO and GAO activity. GAOs exhibited a substantially higher biomass decay rate as compared to PAOs under these conditions, which revealed a higher survival capacity for PAOs, representing an advantage for PAOs in EBPR processes. This superior survival capacity of PAOs under conditions more closely resembling a full-scale environment was linked with their ability to maintain a residual level of PHA reserves for longer than GAOs, providing them with an effective energy source for aerobic maintenance processes. Overall, this work shows that each of these key operational conditions play an important role in the PAO-GAO competition and should be considered in WWTP models in order to improve EBPR processes.

Comparison between blue and green stimulated luminescence of Al(2)O(3):C

This paper presents a systematic comparison of OSL signals from Al(2)O(3):C when stimulated with blue and green light. Al(2)O(3):C detectors were irradiated with various doses and submitted to various bleaching regimes using yellow, green and blue light. Most of the investigations were carried out using Luxel (TM)-type detectors used in the commercial Luxet (TM) and InLight (TM) dosimetry systems (Landauer Inc.). Al(2)O(3):C single crystals and Al(2)O(3):C powder were also used to complement the investigations. The results show that, although blue stimulation provides faster readout times (OSL curves that decayed faster) and higher initial OSL intensity than green stimulation, blue stimulation introduced complicating factors. These include incomplete bleaching of the dosimetric trap when the Al(2)O(3):C detectors are bleached with yellow or green light and the OSL is recorded with blue light stimulation, and an increased residual level due to stimulation of charge carriers from deep traps. The results warrant caution when using blue stimulation to measure the OSL signal from Al(2)O(3):C detectors, particularly if the doses involved are low and the detectors have been previously exposed to high doses. (C) 2010 Elsevier Ltd. All rights reserved.

Iterative reconstruction as a novel method of radiation dose reduction at computed tomography in patients with Crohn's disease

Prior work of our research group, that quantified the alarming levels of radiation dose to patients with Crohn’s disease from medical imaging and the notable shift towards CT imaging making these patients an at risk group, provided context for this work. CT delivers some of the highest doses of ionising radiation in diagnostic radiology. Once a medical imaging examination is deemed justified, there is an onus on the imaging team to endeavour to produce diagnostic quality CT images at the lowest possible radiation dose to that patient. The fundamental limitation with conventional CT raw data reconstruction was the inherent coupling of administered radiation dose with observed image noise – the lower the radiation dose, the noisier the image. The renaissance, rediscovery and refinement of iterative reconstruction removes this limitation allowing either an improvement in image quality without increasing radiation dose or maintenance of image quality at a lower radiation dose compared with traditional image reconstruction. This thesis is fundamentally an exercise in optimisation in clinical CT practice with the objectives of assessment of iterative reconstruction as a method for improvement of image quality in CT, exploration of the associated potential for radiation dose reduction, and development of a new split dose CT protocol with the aim of achieving and validating diagnostic quality submillisiever t CT imaging in patients with Crohn’s disease. In this study, we investigated the interplay of user-selected parameters on radiation dose and image quality in phantoms and cadavers, comparing traditional filtered back projection (FBP) with iterative reconstruction algorithms. This resulted in the development of an optimised, refined and appropriate split dose protocol for CT of the abdomen and pelvis in clinical patients with Crohn’s disease allowing contemporaneous acquisition of both modified and conventional dose CT studies. This novel algorithm was then applied to 50 patients with a suspected acute complication of known Crohn’s disease and the raw data reconstructed with FBP, adaptive statistical iterative reconstruction (ASiR) and model based iterative reconstruction (MBIR). Conventional dose CT images with FBP reconstruction were used as the reference standard with which the modified dose CT images were compared in terms of radiation dose, diagnostic findings and image quality indices. As there are multiple possible user-selected strengths of ASiR available, these were compared in terms of image quality to determine the optimal strength for this modified dose CT protocol. Modified dose CT images with MBIR were also compared with contemporaneous abdominal radiograph, where performed, in terms of diagnostic yield and radiation dose. Finally, attenuation measurements in organs, tissues, etc. with each reconstruction algorithm were compared to assess for preservation of tissue characterisation capabilities. In the phantom and cadaveric models, both forms of iterative reconstruction examined (ASiR and MBIR) were superior to FBP across a wide variety of imaging protocols, with MBIR superior to ASiR in all areas other than reconstruction speed. We established that ASiR appears to work to a target percentage noise reduction whilst MBIR works to a target residual level of absolute noise in the image. Modified dose CT images reconstructed with both ASiR and MBIR were non-inferior to conventional dose CT with FBP in terms of diagnostic findings, despite reduced subjective and objective indices of image quality. Mean dose reductions of 72.9-73.5% were achieved with the modified dose protocol with a mean effective dose of 1.26mSv. MBIR was again demonstrated superior to ASiR in terms of image quality. The overall optimal ASiR strength for the modified dose protocol used in this work is ASiR 80%, as this provides the most favourable balance of peak subjective image quality indices with less objective image noise than the corresponding conventional dose CT images reconstructed with FBP. Despite guidelines to the contrary, abdominal radiographs are still often used in the initial imaging of patients with a suspected complication of Crohn’s disease. We confirmed the superiority of modified dose CT with MBIR over abdominal radiographs at comparable doses in detection of Crohn’s disease and non-Crohn’s disease related findings. Finally, we demonstrated (in phantoms, cadavers and in vivo) that attenuation values do not change significantly across reconstruction algorithms meaning preserved tissue characterisation capabilities with iterative reconstruction. Both adaptive statistical and model based iterative reconstruction algorithms represent feasible methods of facilitating acquisition diagnostic quality CT images of the abdomen and pelvis in patients with Crohn’s disease at markedly reduced radiation doses. Our modified dose CT protocol allows dose savings of up to 73.5% compared with conventional dose CT, meaning submillisievert imaging is possible in many of these patients.

Intensity dependent residual amplitude modulation in electro-optic phase modulators

Residual amplitude modulation (RAM) mechanisms in electro-optic phase modulators are detrimental in applications that require high purity phase modulation of the incident laser beam. While the origins of RAMare not fully understood, measurements have revealed that it depends on the beam properties of the laser as well as the properties of the medium. Here we present experimental and theoretical results that demonstrate, for the first time, the dependence of RAM production in electro-optic phase modulators on beam intensity. The results show an order of magnitude increase in the level of RAM, around 10 dB, with a fifteenfold enhancement in the input intensity from 12 to 190 mW/mm 2. We show that this intensity dependent RAM is photorefractive in origin. © 2012 Optical Society of America.

Reducing residual amplitude modulation in electro-optic phase modulators by erasing photorefractive scatter

Residual amplitude modulation (RAM) is an unwanted noise source in electro-optic phase modulators. The analysis presented shows that while the magnitude of the RAM produced by a MgO:LiNbO3 modulator increases with intensity, its associated phase becomes less well defined. This combination results in temporal fluctuations in RAM that increase with intensity. This behaviour is explained by the presented phenomenological model based on gradually evolving photorefractive scattering centres randomly distributed throughout the optically thick medium. This understanding is exploited to show that RAM can be reduced to below the 10-5 level by introducing an intense optical beam to erase the photorefractive scatter.

Dependence of residual amplitude noise in electro-optic phase modulators on the intensity distribution of the incident field

Our results demonstrate that photorefractive residual amplitude modulation (RAM) noise in electro-optic modulators (EOMs) can be reduced by modifying the incident beam intensity distribution. Here we report an order of magnitude reduction in RAM when beams with uniform intensity (flat-top) profiles, generated with an LCOS-SLM, are used instead of the usual fundamental Gaussian mode (TEM00). RAM arises from the photorefractive amplified scatter noise off the defects and impurities within the crystal. A reduction in RAM is observed with increasing intensity uniformity (flatness), which is attributed to a reduction in space charge field on the beam axis. The level of RAM reduction that can be achieved is physically limited by clipping at EOM apertures, with the observed results agreeing well with a simple model. These results are particularly important in applications where the reduction of residual amplitude modulation to 10^-6 is essential.

A neutron diffraction investigation of residual stresses in rail ends after severe deformation of rail surfaces

Insulated rail joints (IRJs) are a primary component of the rail track safety and signalling systems. Rails are supported by two fishplates which are fastened by bolts and nuts and, with the support of sleepers and track ballast, form an integrated assembly. IRJ failure can result from progressive defects, the propagation of which is influenced by residual stresses in the rail. Residual stresses change significantly during service due to the complex deformation and damage effects associated with wheel rolling, sliding and impact. IRJ failures can occur when metal flows over the insulated rail gap (typically 6-8 mm width), breaks the electrically isolated section of track and results in malfunction of the track signalling system. In this investigation, residual stress measurements were obtained from rail-ends which had undergone controlled amounts of surface plastic deformation using a full scale wheel-on-track simulation test rig. Results were compared with those obtained from similar investigations performed on rail ends associated with ex-service IRJs. Residual stresses were measured by neutron diffraction at the Australian Nuclear Science and Technology Organisation (ANSTO). Measurements with constant gauge volume 3x3x3 mm3 were carried in the central vertical plane on 5mm thick sliced rail samples cut by an electric discharge machine (EDM). Stress evolution at the rail ends was found to exhibit characteristics similar to those of the ex-service rails, with a compressive zone of 5mm deep that is counterbalanced by a tension zone beneath, extending to a depth of around 15mm. However, in contrast to the ex-service rails, the type of stress distribution in the test-rig deformed samples was apparently different due to the localization of load under the particular test conditions. In the latter, in contrast with clear stress evolution, there was no obvious evolution of d0. Since d0 reflects rather long-term accumulation of crystal lattice damage and microstructural changes due to service load, the loading history of the test rig samples has not reached the same level as the ex-service rails. It is concluded that the wheel-on-rail simulation rig provides the potential capability for testing the wheel-rail rolling contact conditions in rails, rail ends and insulated rail joints.

CAE-based prediction of projectile residual velocity for impact on single and multi-layered metallic armour plates

The present, paper deals with the CAE-based study Of impact of jacketed projectiles on single- and multi-layered metal armour plates using LS-DYNA. The validation of finite element modelling procedure is mainly based on the mesh convergence study using both shell and solid elements for representing single-layered mild steel target plates. It, is shown that the proper choice of mesh density and the strain rate-dependent material properties are essential for all accurate prediction of projectile residual velocity. The modelling requirements are initially arrived at by correlating against test residual velocities for single-layered mild steel plates of different depths at impact velocities in the ran.-c of approximately 800-870 m/s. The efficacy of correlation is adjudged, in terms of a 'correlation index', defined in the paper: for which values close to unity are desirable. The experience gained for single-layered plates is next; used in simulating projectile impacts on multi-layered mild steel target plates and once again a high degree of correlation with experimental residual velocities is observed. The study is repeated for single- and multi-layered aluminium target plates with a similar level of success in test residual velocity prediction. TO the authors' best knowledge, the present comprehensive study shows in particular for the first time that, with a. proper modelling approach, LS-DYNA can be used with a great degree of confidence in designing perforation-resistant single and multi-layered metallic armour plates.

Effect of residual stresses and metallographic stability on the over all performance of integral diaphragm material

The integral diaphragm pressure transducer consists of a diaphragm machined from precipitation hardened martensitic (APX4) steel. Its performance is quite significant as it depends upon various factors such as mechanical properties including induced residual stress levels, metallurgical and physical parameters due to different stages of processing involved. Hence, the measurement and analysis of residual stress becomes very important from the point of in-service assessment of a component. In the present work, the stress measurements have been done using the X-ray diffraction (XRD) technique, which is a non-destructive test (NDT). This method is more reliable and widely used compared to the other NDT techniques. The metallurgical aspects have been studied by adopting the conventional metallographic practices including examination of microstructure using light microscope. The dimensional measurements have been carried out using dimensional gauge. The results of the present investigation reveals that the diaphragm material after undergoing series of realization processes has yielded good amount of retained austenite in it. Also, the presence of higher compressive stresses induced in the transducer results in non-linearity, zero shift and dimensional instability. The problem of higher retained austenite content and higher compressive stress have been overcome by adopting a new realization process involving machining and cold and hot stabilization soak which has brought down the retained austenite content to about 5–6% and acceptable level of compressive stress in the range −100 to −150 MPa with fine tempered martensitic phase structure and good dimensional stability. The new realization process seems to be quite effective in terms of controlling retained austenite content, residual stress, metallurgical phase as well as dimensional stability and this may result in minimum zero shift of the diaphragm system.

Measurements of adequacy of anesthesia and level of consciousness during surgery and intensive care

The adequacy of anesthesia has been studied since the introduction of balanced general anesthesia. Commercial monitors based on electroencephalographic (EEG) signal analysis have been available for monitoring the hypnotic component of anesthesia from the beginning of the 1990s. Monitors measuring the depth of anesthesia assess the cortical function of the brain, and have gained acceptance during surgical anesthesia with most of the anesthetic agents used. However, due to frequent artifacts, they are considered unsuitable for monitoring consciousness in intensive care patients. The assessment of analgesia is one of the cornerstones of general anesthesia. Prolonged surgical stress may lead to increased morbidity and delayed postoperative recovery. However, no validated monitoring method is currently available for evaluating analgesia during general anesthesia. Awareness during anesthesia is caused by an inadequate level of hypnosis. This rare but severe complication of general anesthesia may lead to marked emotional stress and possibly posttraumatic stress disorder. In the present series of studies, the incidence of awareness and recall during outpatient anesthesia was evaluated and compared with that of in inpatient anesthesia. A total of 1500 outpatients and 2343 inpatients underwent a structured interview. Clear intraoperative recollections were rare the incidence being 0.07% in outpatients and 0.13% in inpatients. No significant differences emerged between outpatients and inpatients. However, significantly smaller doses of sevoflurane were administered to outpatients with awareness than those without recollections (p<0.05). EEG artifacts in 16 brain-dead organ donors were evaluated during organ harvest surgery in a prospective, open, nonselective study. The source of the frontotemporal biosignals in brain-dead subjects was studied, and the resistance of bispectral index (BIS) and Entropy to the signal artifacts was compared. The hypothesis was that in brain-dead subjects, most of the biosignals recorded from the forehead would consist of artifacts. The original EEG was recorded and State Entropy (SE), Response Entropy (RE), and BIS were calculated and monitored during solid organ harvest. SE differed from zero (inactive EEG) in 28%, RE in 29%, and BIS in 68% of the total recording time (p<0.0001 for all). The median values during the operation were SE 0.0, RE 0.0, and BIS 3.0. In four of the 16 organ donors, EEG was not inactive, and unphysiologically distributed, nonreactive rhythmic theta activity was present in the original EEG signal. After the results from subjects with persistent residual EEG activity were excluded, SE, RE, and BIS differed from zero in 17%, 18%, and 62% of the recorded time, respectively (p<0.0001 for all). Due to various artifacts, the highest readings in all indices were recorded without neuromuscular blockade. The main sources of artifacts were electrocauterization, electromyography (EMG), 50-Hz artifact, handling of the donor, ballistocardiography, and electrocardiography. In a prospective, randomized study of 26 patients, the ability of Surgical Stress Index (SSI) to differentiate patients with two clinically different analgesic levels during shoulder surgery was evaluated. SSI values were lower in patients with an interscalene brachial plexus block than in patients without an additional plexus block. In all patients, anesthesia was maintained with desflurane, the concentration of which was targeted to maintain SE at 50. Increased blood pressure or heart rate (HR), movement, and coughing were considered signs of intraoperative nociception and treated with alfentanil. Photoplethysmographic waveforms were collected from the contralateral arm to the operated side, and SSI was calculated offline. Two minutes after skin incision, SSI was not increased in the brachial plexus block group and was lower (38 ± 13) than in the control group (58 ± 13, p<0.005). Among the controls, one minute prior to alfentanil administration, SSI value was higher than during periods of adequate antinociception, 59 ± 11 vs. 39 ± 12 (p<0.01). The total cumulative need for alfentanil was higher in controls (2.7 ± 1.2 mg) than in the brachial plexus block group (1.6 ± 0.5 mg, p=0.008). Tetanic stimulation to the ulnar region of the hand increased SSI significantly only among patients with a brachial plexus block not covering the site of stimulation. Prognostic value of EEG-derived indices was evaluated and compared with Transcranial Doppler Ultrasonography (TCD), serum neuron-specific enolase (NSE) and S-100B after cardiac arrest. Thirty patients resuscitated from out-of-hospital arrest and treated with induced mild hypothermia for 24 h were included. Original EEG signal was recorded, and burst suppression ratio (BSR), RE, SE, and wavelet subband entropy (WSE) were calculated. Neurological outcome during the six-month period after arrest was assessed with the Glasgow-Pittsburgh Cerebral Performance Categories (CPC). Twenty patients had a CPC of 1-2, one patient had a CPC of 3, and nine patients died (CPC 5). BSR, RE, and SE differed between good (CPC 1-2) and poor (CPC 3-5) outcome groups (p=0.011, p=0.011, p=0.008, respectively) during the first 24 h after arrest. WSE was borderline higher in the good outcome group between 24 and 48 h after arrest (p=0.050). All patients with status epilepticus died, and their WSE values were lower (p=0.022). S-100B was lower in the good outcome group upon arrival at the intensive care unit (p=0.010). After hypothermia treatment, NSE and S-100B values were lower (p=0.002 for both) in the good outcome group. The pulsatile index was also lower in the good outcome group (p=0.004). In conclusion, the incidence of awareness in outpatient anesthesia did not differ from that in inpatient anesthesia. Outpatients are not at increased risk for intraoperative awareness relative to inpatients undergoing general anesthesia. SE, RE, and BIS showed non-zero values that normally indicate cortical neuronal function, but were in these subjects mostly due to artifacts after clinical brain death diagnosis. Entropy was more resistant to artifacts than BIS. During general anesthesia and surgery, SSI values were lower in patients with interscalene brachial plexus block covering the sites of nociceptive stimuli. In detecting nociceptive stimuli, SSI performed better than HR, blood pressure, or RE. BSR, RE, and SE differed between the good and poor neurological outcome groups during the first 24 h after cardiac arrest, and they may be an aid in differentiating patients with good neurological outcomes from those with poor outcomes after out-of-hospital cardiac arrest.

Fuzzy-logic-based health monitoring and residual-life prediction for composite helicopter rotor

A health-monitoring and life-estimation strategy for composite rotor blades is developed in this work. The cross-sectional stiffness reduction obtained by physics-based models is expressed as a function of the life of the structure using a recent phenomenological damage model. This stiffness reduction is further used to study the behavior of measurable system parameters such as blade deflections, loads, and strains of a composite rotor blade in static analysis and forward flight. The simulated measurements are obtained using an aeroelastic analysis of the composite rotor blade based on the finite element in space and time with physics-based damage modes that are then linked to the life consumption of the blade. The model-based measurements are contaminated with noise to simulate real data. Genetic fuzzy systems are developed for global online prediction of physical damage and life consumption using displacement- and force-based measurement deviations between damaged and undamaged conditions. Furthermore, local online prediction of physical damage and life consumption is done using strains measured along the blade length. It is observed that the life consumption in the matrix-cracking zone is about 12-15% and life consumption in debonding/delamination zone is about 45-55% of the total life of the blade. It is also observed that the success rate of the genetic fuzzy systems depends upon the number of measurements, type of measurements and training, and the testing noise level. The genetic fuzzy systems work quite well with noisy data and are recommended for online structural health monitoring of composite helicopter rotor blades.

Residual noise covariance for Planck low-resolution data analysis

Aims: Develop and validate tools to estimate residual noise covariance in Planck frequency maps. Quantify signal error effects and compare different techniques to produce low-resolution maps. Methods: We derive analytical estimates of covariance of the residual noise contained in low-resolution maps produced using a number of map-making approaches. We test these analytical predictions using Monte Carlo simulations and their impact on angular power spectrum estimation. We use simulations to quantify the level of signal errors incurred in different resolution downgrading schemes considered in this work. Results: We find an excellent agreement between the optimal residual noise covariance matrices and Monte Carlo noise maps. For destriping map-makers, the extent of agreement is dictated by the knee frequency of the correlated noise component and the chosen baseline offset length. The significance of signal striping is shown to be insignificant when properly dealt with. In map resolution downgrading, we find that a carefully selected window function is required to reduce aliasing to the sub-percent level at multipoles, ell > 2Nside, where Nside is the HEALPix resolution parameter. We show that sufficient characterization of the residual noise is unavoidable if one is to draw reliable contraints on large scale anisotropy. Conclusions: We have described how to compute the low-resolution maps, with a controlled sky signal level, and a reliable estimate of covariance of the residual noise. We have also presented a method to smooth the residual noise covariance matrices to describe the noise correlations in smoothed, bandwidth limited maps.