Subgroup Determination of Bulgarian Isolates of Cucumber Mosaic Virus and the Presence of Satellite RNAs.
Cucumber mosaic virus (CMV) is one of the most important viruses in Bulgaria, causing severe losses to agriculture, but little is known about the occurrence and distribution of subgroups within the country or the presence of satellite RNAs (satRNAs). Samples showing typical symptoms (mild to severe mosaic, vein clearing, vein necrosis, leaf deformation, stunting, and fruit necrosis) on several important crops (tomato, cucumber, pepper, bean, courgette, and tobacco) were collected from the main agricultural regions of the country. Isolates were maintained by sap inoculation in tobacco plants. Total RNAs were isolated from 38 samples (including two from bean) and used in reverse transcription-polymerase chain reaction (RT-PCR) assay with primers corresponding to the coat protein (CP) gene of RNA3 (3). A single strong band, 870 bp in length, was produced from all these samples. Amplified products were analyzed for subgroup differentiation by digestion with the restriction endonucleases MspI (3), PvuII, and EcoRI. The MspI subgroups 2 and 1 designated by Rizos et al. (3) according to their restriction endonuclease digest data correspond to the subgroups I and II widely used in the literature and based on serology, sequence data, and other properties. In this report, the subgroups are referred to as I and II for the sake of clarity. Isolates in both subgroups were found in all the main regions of Bulgaria. A few variations in MspI and EcoRI digestion patterns were seen, indicating some variability between isolates within subgroups. Only five samples, three from tomato and two from pepper, were found to be subgroup II. Subgroup I isolates were found in all the crops sampled. The PCR product from one representative isolate of each subgroup was cloned and sequenced by standard procedures. Alignment of the nucleotide and predicted amino acid sequences with published sequences of the CMV CP gene confirmed that the amplified products were derived from CMV. A further eight samples from bean gave only weak amplification and digestion of the products suggested they were likely to be subgroup II. However, these samples were unusual in not inducing symptoms in inoculated tobacco and in being difficult to propagate. The nature of these virus isolates is therefore unclear. Only a single occurrence in Bulgaria of satRNA of CMV has been reported (4) but in this study satRNAs were detected by RT-PCR (1) in total plant RNA extracts of 21 of the 38 samples tested. Amplified products of two of them, NB and 146D, were sequenced; comparison with published sequences confirmed that they were derived from CMV satellite. As expected from the symptoms induced by these isolates, a sequence homologous to the domain of satRNA Y responsible for bright yellow mosaic on tobacco (2) was identified in satRNA NB but not in satRNA 146D. satRNAs were not detected in the eight bean samples that had given only weak amplification with the CMV CP gene primers. The results presented here clearly demonstrate the presence of both subgroups of CMV in Bulgaria. Although CMV in Bulgaria has been studied previously by serological methods, no evidence had been found for the presence of subgroup II. References: (1) F. Grieco et al. Virology 229:166, 1997. (2) C. Masuta and Y. Takanami. Plant Cell 1:1165, 1989. (3) H. Rizos et al. J. Gen. Virol. 73:2099, 1992. (4) E. Stoimenova. J. Cult. Collect. 1:45, 1995.


Efficacy and safety of levoketoconazole in the treatment of endogenous Cushing's syndrome (SONICS): a phase 3, multicentre, open-label, single-arm trial.
Levoketoconazole is a ketoconazole stereoisomer in development for treatment of Cushing's syndrome and has not been assessed previously in a clinical trial in patients with Cushing's syndrome. We aimed to investigate the efficacy and safety of levoketoconazole in patients with endogenous Cushing's syndrome. SONICS is a phase 3, multicentre, open-label, non-randomised, single-arm study in which we recruited adults (≥18 years) with confirmed Cushing's syndrome and a mean 24-h urinary free cortisol (mUFC) of at least 1·5 times the upper limit of normal from 60 hospital and community sites in 19 countries (15 countries in Europe, and Canada, Israel, Turkey, and the USA). Patients were treated with oral levoketoconazole in a 2-21 week incremental dose-titration phase starting at 150 mg twice daily (150 mg increments until mUFC normalisation, maximum 600 mg twice daily) and a 6-month maintenance phase. The primary outcome was the proportion of patients with mUFC normalisation at end of maintenance, without dose increase during the maintenance phase (in the intention-to-treat population). Prespecified adverse events of special interest were potential liver toxicity, corrected QT prolongation, and adrenal insufficiency. This trial is registered with ClinicalTrials.gov, NCT01838551. Between July 30, 2014, and June 30, 2017, 201 individuals were screened and 94 patients were enrolled and received at least one dose of study medication. Of the 94 patients, 80 (85%) had pituitary Cushing's syndrome. Mean mUFC at baseline was 671·4 nmol/24 h (243·3 μg/24 h), which is 4·9 times the upper limit of normal. Of the 77 patients who advanced to the maintenance phase, 62 (81%) had mUFC normalisation by end-of-dose titration. At the end of the 6-month maintenance phase, 29 (31%) of 94 patients were responders; the least-squares mean estimate of the proportion of responders was 0·30 (95% CI 0·21-0·40; p=0·0154 vs null hypothesis of ≤0·20). The most common adverse events in the 94 patients were nausea (30 [32%]) and headache (26 [28%]). Adverse events led to study discontinuation in 12 (13%) of 94 patients. Two patients had a QT interval (Fridericia corrected) of more than 500 ms, and three patients had suspected adrenal insufficiency. Alanine aminotransferase reversibly increased to more than three times the upper limit of normal in ten (11%) patients. Four patients had serious adverse events that were considered probably or definitely related to the study drug: abnormal liver function test results (n=1), prolonged QT interval (n=2), and adrenal insufficiency (n=1). One person died from colon carcinoma unrelated to study medication. Twice-daily oral levoketoconazole treatment led to sustained improvements in urinary free cortisol, with an acceptable safety and tolerability profile. Levoketoconazole might represent a useful therapeutic option for the medical treatment of Cushing's syndrome. Strongbridge Biopharma.


Myotonic dystrophy 1 in the nervous system: from the clinic to molecular mechanisms.
Myotonic dystrophy type 1 (DM1) is a dominant neuromuscular disorder caused by the expansion of trinucleotide CTG repeats in the 3'-untranslated region (3'-UTR) of the DMPK gene. Prominent features of classical DM1 are muscle wasting and myotonia, whereas mental retardation is distinctive for congenital DM1. The main nervous system symptoms of DM1 are cognitive impairment, neuroendocrine dysfunction, and personality and behavior abnormalities. It is thought that expansion of CTG repeats causes DM1 pathology through different molecular mechanisms; however, a growing body of evidence indicates that an RNA gain-of-function mechanism plays a major role in the disease development. At the skeletal muscle level, three main molecular events can be distinguished in this model: 1) formation of nuclear foci that are composed at least of mutant DMPK mRNA and recruited RNA-binding proteins, such as splicing regulators and transcription factors; 2) disturbance of alternative splicing of specific genes; and 3) impairment of cell differentiation. Contrasting with the substantial advances in understanding DM1 muscle pathology, the molecular basis of DM1 in the nervous system has just started to be revealed. This review focuses in the DM1 nervous system pathology and provides an overview of the genetic and molecular studies analyzing the effects of the DMPK gene CUG expanded repeats on cell function in neuronal systems. A comparison between the molecular mechanisms of DM1 in the skeletal muscle and those identified in DM1 nervous system models is provided. Finally, future directions in the study of DM1 in the nervous system are discussed.


Overweight, obesity, and risk of cardiometabolic multimorbidity: pooled analysis of individual-level data for 120 813 adults from 16 cohort studies from the USA and Europe.
Although overweight and obesity have been studied in relation to individual cardiometabolic diseases, their association with risk of cardiometabolic multimorbidity is poorly understood. Here we aimed to establish the risk of incident cardiometabolic multimorbidity (ie, at least two from: type 2 diabetes, coronary heart disease, and stroke) in adults who are overweight and obese compared with those who are a healthy weight. We pooled individual-participant data for BMI and incident cardiometabolic multimorbidity from 16 prospective cohort studies from the USA and Europe. Participants included in the analyses were 35 years or older and had data available for BMI at baseline and for type 2 diabetes, coronary heart disease, and stroke at baseline and follow-up. We excluded participants with a diagnosis of diabetes, coronary heart disease, or stroke at or before study baseline. According to WHO recommendations, we classified BMI into categories of healthy (20·0-24·9 kg/m2), overweight (25·0-29·9 kg/m2), class I (mild) obesity (30·0-34·9 kg/m2), and class II and III (severe) obesity (≥35·0 kg/m2). We used an inclusive definition of underweight (<20 kg/m2) to achieve sufficient case numbers for analysis. The main outcome was cardiometabolic multimorbidity (ie, developing at least two from: type 2 diabetes, coronary heart disease, and stroke). Incident cardiometabolic multimorbidity was ascertained via resurvey or linkage to electronic medical records (including hospital admissions and death). We analysed data from each cohort separately using logistic regression and then pooled cohort-specific estimates using random-effects meta-analysis. Participants were 120 813 adults (mean age 51·4 years, range 35-103; 71 445 women) who did not have diabetes, coronary heart disease, or stroke at study baseline (1973-2012). During a mean follow-up of 10·7 years (1995-2014), we identified 1627 cases of multimorbidity. After adjustment for sociodemographic and lifestyle factors, compared with individuals with a healthy weight, the risk of developing cardiometabolic multimorbidity in overweight individuals was twice as high (odds ratio [OR] 2·0, 95% CI 1·7-2·4; p<0·0001), almost five times higher for individuals with class I obesity (4·5, 3·5-5·8; p<0·0001), and almost 15 times higher for individuals with classes II and III obesity combined (14·5, 10·1-21·0; p<0·0001). This association was noted in men and women, young and old, and white and non-white participants, and was not dependent on the method of exposure assessment or outcome ascertainment. In analyses of different combinations of cardiometabolic conditions, odds ratios associated with classes II and III obesity were 2·2 (95% CI 1·9-2·6) for vascular disease only (coronary heart disease or stroke), 12·0 (8·1-17·9) for vascular disease followed by diabetes, 18·6 (16·6-20·9) for diabetes only, and 29·8 (21·7-40·8) for diabetes followed by vascular disease. The risk of cardiometabolic multimorbidity increases as BMI increases; from double in overweight people to more than ten times in severely obese people compared with individuals with a healthy BMI. Our findings highlight the need for clinicians to actively screen for diabetes in overweight and obese patients with vascular disease, and pay increased attention to prevention of vascular disease in obese individuals with diabetes. NordForsk, Medical Research Council, Cancer Research UK, Finnish Work Environment Fund, and Academy of Finland.


Characterisation of Development and Electrophysiological Mechanisms Underlying Rhythmicity of the Avian Lymph Heart.
Despite significant advances in tissue engineering such as the use of scaffolds, bioreactors and pluripotent stem cells, effective cardiac tissue engineering for therapeutic purposes has remained a largely intractable challenge. For this area to capitalise on such advances, a novel approach may be to unravel the physiological mechanisms underlying the development of tissues that exhibit rhythmic contraction yet do not originate from the cardiac lineage. Considerable attention has been focused on the physiology of the avian lymph heart, a discrete organ with skeletal muscle origins yet which displays pacemaker properties normally only found in the heart. A functional lymph heart is essential for avian survival and growth in ovo. The histological nature of the lymph heart is similar to skeletal muscle although molecular and bioelectrical characterisation during development to assess mechanisms that contribute towards lymph heart contractile rhythmicity have not been undertaken. A better understanding of these processes may provide exploitable insights for therapeutic rhythmically contractile tissue engineering approaches in this area of significant unmet clinical need. Here, using molecular and electrophysiological approaches, we describe the molecular development of the lymph heart to understand how this skeletal muscle becomes fully functional during discrete in ovo stages of development. Our results show that the lymph heart does not follow the normal transitional programme of myogenesis as documented in most skeletal muscle, but instead develops through a concurrent programme of precursor expansion, commitment to myogenesis and functional differentiation which offers a mechanistic explanation for its rapid development. Extracellular electrophysiological field potential recordings revealed that the peak-to-peak amplitude of electrically evoked local field potentials elicited from isolated lymph heart were significantly reduced by treatment with carbachol; an effect that could be fully reversed by atropine. Moreover, nifedipine and cyclopiazonic acid both significantly reduced peak-to-peak local field potential amplitude. Optical recordings of lymph heart showed that the organ's rhythmicity can be blocked by the HCN channel blocker, ZD7288; an effect also associated with a significant reduction in peak-to-peak local field potential amplitude. Additionally, we also show that isoforms of HCN channels are expressed in avian lymph heart. These results demonstrate that cholinergic signalling and L-type Ca2+ channels are important in excitation and contraction coupling, while HCN channels contribute to maintenance of lymph heart rhythmicity.


Concise review: mesenchymal stem/multipotent stromal cells: the state of transdifferentiation and modes of tissue repair--current views.
Mesenchymal stem cells or multipotent stromal cells (MSCs) isolated from the bone marrow of adult organisms were initially characterized as plastic adherent, fibroblastoid cells with the capacity to generate heterotopic osseous tissue when transplanted in vivo. In recent years, MSCs or MSC-like cells have been shown to reside within the connective tissue of most organs, and their surface phenotype has been well described. A large number of reports have also indicated that the cells possess the capacity to transdifferentiate into epithelial cells and lineages derived from the neuroectoderm. The broad developmental plasticity of MSCs was originally thought to contribute to their demonstrated efficacy in a wide variety of experimental animal models of disease as well as in human clinical trials. However, new findings suggest that the ability of MSCs to alter the tissue microenvironment via secretion of soluble factors may contribute more significantly than their capacity for transdifferentiation in tissue repair. Herein, we critically evaluate the literature describing the plasticity of MSCs and offer insight into how the molecular and functional heterogeneity of this cell population, which reflects the complexity of marrow stroma as an organ system, may confound interpretation of their transdifferentiation potential. Additionally, we argue that this heterogeneity also provides a basis for the broad therapeutic efficacy of MSCs.


Whole grains, legumes, and the subsequent meal effect: implications for blood glucose control and the role of fermentation.
Whole grains and legumes are known to reduce postprandial glycemia and, in some instances, insulinemia. However, the subsequent meal effect of ingesting whole grains and legumes is less well known. That is, inclusion of whole grains or legumes at breakfast decreases postprandial glycemia at lunch and/or dinner on the same day whereas consumption of a whole grain or lentil dinner reduces glycemia at breakfast the following morning. This effect is lost upon milling, processing, and cooking at high temperatures. The subsequent meal effect has important implications for the control of day-long blood glucose, and may be partly responsible for the reduction in diabetes incidence associated with increased whole grain and legume intake. This paper describes the subsequent meal effect and explores the role of acute glycemia, presence of resistant starch, and fermentation of indigestible carbohydrate as the mechanisms responsible for this effect.


A facile method for the detection of DNA by using gold nanoparticle probes coupled with dynamic light scattering.
In this paper, we present a simple and rapid method for deoxyribonucleic acid (DNA) detection using gold nanoparticle probes coupled with dynamic light scattering (DLS) analysis. The redox agent 1,4-dithio-dl-threitol cross-links the gold nanoparticles (AuNPs) to form clusters, while the monothiol DNA could terminate the formation and stabilize the assembled clusters by their negatively charge-based repulsions. By varying the concentration of DNA, the different sizes of DNA-AuNP clusters can be obtained. The sizes of the DNA-AuNP clusters were determined by DLS. A linear correlation was obtained between the sizes and the logarithm of DNA concentration from 2 nM to 5 μM with a detection limit of 1 nM (S/N = 3).


A Paleolithic diet confers higher insulin sensitivity, lower C-reactive protein and lower blood pressure than a cereal-based diet in domestic pigs.
A Paleolithic diet has been suggested to be more in concordance with human evolutionary legacy than a cereal based diet. This might explain the lower incidence among hunter-gatherers of diseases of affluence such as type 2 diabetes, obesity and cardiovascular disease. The aim of this study was to experimentally study the long-term effect of a Paleolithic diet on risk factors for these diseases in domestic pigs. We examined glucose tolerance, post-challenge insulin response, plasma C-reactive protein and blood pressure after 15 months on Paleolithic diet in comparison with a cereal based swine feed. Upon weaning twenty-four piglets were randomly allocated either to cereal based swine feed (Cereal group) or cereal free Paleolithic diet consisting of vegetables, fruit, meat and a small amount of tubers (Paleolithic group). At 17 months of age an intravenous glucose tolerance test was performed and pancreas specimens were collected for immunohistochemistry. Group comparisons of continuous variables were made by use of the t-test. P < 0.05 was chosen for statistical significance. Simple and multivariate correlations were evaluated by use of linear regression analysis. At the end of the study the Paleolithic group weighed 22% less and had 43% lower subcutaneous fat thickness at mid sternum. No significant difference was seen in fasting glucose between groups. Dynamic insulin sensitivity was significantly higher (p = 0.004) and the insulin response was significantly lower in the Paleolithic group (p = 0.001). The geometric mean of C-reactive protein was 82% lower (p = 0.0007) and intra-arterial diastolic blood pressure was 13% lower in the Paleolithic group (p = 0.007). In evaluations of multivariate correlations, diet emerged as the strongest explanatory variable for the variations in dynamic insulin sensitivity, insulin response, C-reactive protein and diastolic blood pressure when compared to other relevant variables such as weight and subcutaneous fat thickness at mid sternum. There was no obvious immunohistochemical difference in pancreatic islets between the groups, but leukocytes were clearly more frequent in sampled pancreas from the Cereal group. This study in domestic pigs suggests that a Paleolithic diet conferred higher insulin sensitivity, lower C-reactive protein and lower blood pressure when compared to a cereal based diet.


Who mixes with whom among men who have sex with men? Implications for modelling the HIV epidemic in southern India.
In India, the identity of men who have sex with men (MSM) is closely related to the role taken in anal sex (insertive, receptive or both), but little is known about sexual mixing between identity groups. Both role segregation (taking only the insertive or receptive role) and the extent of assortative (within-group) mixing are known to affect HIV epidemic size in other settings and populations. This study explores how different possible mixing scenarios, consistent with behavioural data collected in Bangalore, south India, affect both the HIV epidemic, and the impact of a targeted intervention. Deterministic models describing HIV transmission between three MSM identity groups (mostly insertive Panthis/Bisexuals, mostly receptive Kothis/Hijras and versatile Double Deckers), were parameterised with behavioural data from Bangalore. We extended previous models of MSM role segregation to allow each of the identity groups to have both insertive and receptive acts, in differing ratios, in line with field data. The models were used to explore four different mixing scenarios ranging from assortative (maximising within-group mixing) to disassortative (minimising within-group mixing). A simple model was used to obtain insights into the relationship between the degree of within-group mixing, R0 and equilibrium HIV prevalence under different mixing scenarios. A more complex, extended version of the model was used to compare the predicted HIV prevalence trends and impact of an HIV intervention when fitted to data from Bangalore. With the simple model, mixing scenarios with increased amounts of assortative (within-group) mixing tended to give rise to a higher R0 and increased the likelihood that an epidemic would occur. When the complex model was fit to HIV prevalence data, large differences in the level of assortative mixing were seen between the fits identified using different mixing scenarios, but little difference was projected in future HIV prevalence trends. An oral pre-exposure prophylaxis (PrEP) intervention was modelled, targeted at the different identity groups. For intervention strategies targeting the receptive or receptive and versatile MSM together, the overall impact was very similar for different mixing patterns. However, for PrEP scenarios targeting insertive or versatile MSM alone, the overall impact varied considerably for different mixing scenarios; more impact was achieved with greater levels of disassortative mixing.


Molecular Characterization of Natural Hybrids of Phytophthora nicotianae and P. cactorum.
ABSTRACT Hybrid isolates of Phytophthora nicotianae x P. cactorum from five different hosts (Cyclamen, Lavandula, Lewisia, Primula, and Spathiphyllum spp.) were identified by their atypical morphology and their well-defined heterozygous isozyme patterns. The hybrid nature of these isolates was tested by restriction fragment length polymorphism analysis of the internal transcribed spacer (ITS) region of rDNA, generating fragments typical for both P. nicotianae and P. cactorum. In hybrid isolates, polymerase chain reactions (PCR) with primers derived from unique parts of the ITS region (ITS-PCR) of both species yielded a combination of unique amplicons typical of both parental species. Eleven hybrid isolates, three isolates of each parental species and two atypical isolates from Rhododendron and Idesia spp. close to P. cactorum, were analyzed for amplified fragment length polymorphisms (AFLP). Consistent differences in AFLP patterns existed among the hybrid isolates, strongly indicating that these hybrids have arisen from independent hybridization events between P. nicotianae and P. cactorum. The two atypical isolates morphologically resembling P. cactorum were identical to the latter species in ITS-restriction fragment length polymorphism and response to the specific PCR primers but were intermediate between P. nicotianae x P. cactorum and P. cactorum in isozyme profiles and AFLP patterns. Since the introduction of hydroponic systems in greenhouses in the Netherlands, outbreaks of Phytophthora diseases are occurring in previously unaffected host species. This may be due to interspecific hybridization events resulting in novel pathogenic behavior.


Stabilization by harmonic intensities for the output signal in heterodyne detection.
We experimentally demonstrated that the technique to stabilize the output signal by harmonic intensities is useful in heterodyne detection with an incoherent light source such as a halogen lamp. The relation between the relative standard deviation of an output signal and the fluctuation of the light intensity is analyzed and simulated. Using the fundamental to sixth harmonics increases the stabilities of output signal approximately 3 times, and subtracting the relative standard deviation of the intensity of light source enhances the stabilities 49 times. The fluctuating phase that is due to the fluctuating frequency and temperature and its power spectrum density for an interferometer is also calculated with the Allan variance.


Which intervention is better for malaria vector control: insecticide mixture long-lasting insecticidal nets or standard pyrethroid nets combined with indoor residual spraying?
Malaria control today is threatened by widespread insecticide resistance in vector populations. The World Health Organization (WHO) recommends the use of a mixture of unrelated insecticides for indoor residual spraying (IRS) and long-lasting insecticidal nets (LNs) or as a combination of interventions for improved vector control and insecticide resistance management. Studies investigating the efficacy of these different strategies are necessary. The efficacy of Interceptor® G2 LN, a newly developed LN treated with a mixture of chlorfenapyr (a pyrrole) and alpha-cypermethrin (a pyrethroid), was compared to a combined chlorfenapyr IRS and Interceptor® LN (a standard alpha-cypermethrin LN) intervention in experimental huts in Cove Southern Benin, against wild, free-flying, pyrethroid-resistant Anopheles gambiae s.l. A direct comparison was also made with a pyrethroid-only net (Interceptor® LN) alone and chorfenapyr IRS alone. WHO resistance bioassays performed during the trial demonstrated a pyrethroid resistance frequency of >90% in the wild An. gambiae s.l. from the Cove hut site. Mortality in the control (untreated net) hut was 5%. Mortality with Interceptor® LN (24%) was lower than with chlorfenapyr IRS alone (59%, P < 0.001). The combined Interceptor® LN and chlorfenapyr IRS intervention and the mixture net (Interceptor® G2 LN) provided significantly higher mortality rates (73 and 76%, respectively) and these did not differ significantly between both treatments (P = 0.15). Interceptor LN induced 46% blood-feeding inhibition compared to the control untreated net, while chlorfenapyr IRS alone provided none. Both mixture/combination strategies also induced substantial levels of blood-feeding inhibition (38% with combined interventions and 30% with Interceptor® G2 LN). A similar trend of improved mortality of pyrethroid-resistant An. gambiae s.l. from Cove was observed with Interceptor® G2 LN (79%) compared to Interceptor LN (42%, P < 0.001) in WHO tunnel tests. The use of chlorfenapyr and alpha-cypermethrin together as a mixture on nets (Interceptor® G2 LN) or a combined chlorfenapyr IRS and pyrethroid LN intervention provides improved control of pyrethroid-resistant malaria vectors by inducing significantly higher levels of mortality through the chlorfenapyr component and providing personal protection through the pyrethroid component. Both strategies are comparable in their potential to improve the control of malaria transmitted by pyrethroid resistant mosquito vectors.


Generation of high energy, 30 fs pulses at 527 nm by hollow-fiber compression technique.
The compression of 300-fs-long, chirp-free laser pulses at 527 nm down to 30 fs is reported. The laser pulses, originated from a frequency-doubled, mode-locked Nd:glass laser, were compressed by a 0.7-m-long, 150-microm-bore-diameter, argon-filled hollow fiber, and a pair of SF10 prisms with a final energy of 160 microJ. These are the shortest, high energy pulses ever produced by direct pulse compression at the central wavelength of 527 nm. The spectral broadening of the pulses propagating inside the hollow fiber was experimentally examined for various filling-gas pressures and input pulse energies. The spectral width of the pulses was broadened up to 25 nm, and 27 nm for argon- and krypton-filled hollow fiber, respectively, at a gas pressure lower than 2 bar. The physical limitations of the hollow-fiber pulse compression technique applied in the visible range are also studied.


Factors Statistically Predicting At-Risk/Problematic Internet Use in a Sample of Young Adolescent Boys and Girls in South Korea.
Aims: This study aimed to investigate in a gender-sensitive manner factors related to at-risk/problematic Internet use (ARPIU) in a sample of young Korean adolescents. Given prior findings, we hypothesized we would observe specific temperamental, social and biological measures that would statistically predict ARPIU in boys and girls, respectively. Method: Subjects included 653 middle-school students from Chuncheon, Korea who completed measures assessing Internet addiction, mood, temperament, and social interactions. Finger digit (2D:4D) ratios were also assessed. Chi-square and logistic regression models were conducted. Results: Among boys and girls, the ARPIU and non-ARPIU groups showed differences in temperament, mood, social tendencies, and gaming behaviors. In boys, IAT correlated inversely with the 2D:4D digit ratio and novelty-seeking and positively with reward-dependence scores when controlling for BDI scores; these relationships were not found in girls. Multivariate analyses showed that among boys, novelty-seeking, harm avoidance, self-transcendence, and daily time spent gaming statistically predicted ARPIU. Among girls, daily time spent gaming, number of best friends, self-directedness, and cooperation statistically predicted ARPIU. Conclusion: ARPIU was linked to specific temperamental, behavioral and biological characteristics, with specific relationships observed in boys and girls. Specific risk factors may exist for boys and girls with respect to their propensities to developing ARPIU, suggesting the need for gender-sensitive approaches to prevent ARPIU in youth.


Evaluation of total oxidative stress and antioxidant capacity of brain tumour patients attending referral hospitals in Addis Ababa, 2020: a comparative cross-sectional study.
The exact cause of brain tumours is still unknown, but disruptions of redox balance are thought to play a significant role in all stages of brain tumour development. However, the roles of free radical imbalance at different grades of brain tumour and degree of oxidative stress before and after surgery have not been addressed in prior studies. A comparative cross-sectional study was conducted to assess the redox imbalance among confirmed brain tumour patients. An institution-based comparative cross-sectional study was conducted on a total of 100 participants (50 brain tumour patients and 50 controls) at referral hospitals in Addis Ababa, Ethiopia. Descriptive statistics, t-test and analysis of variance (ANOVA) (post-hoc) analysis were used and statistical significance was declared at p ≤ 0.05. The serum oxidised glutathione and total oxidative stress were significantly higher in the serum of brain tumour patients (0.72 ± 0.03 μM/μg and 9.66 ± 1.76 μmol H2O2 Eq/L, respectively) compared to the control group (0.21 ± 0.07 μM/μg and 6.59 ± 0.81 μmol H2O2 Eq/L, respectively) (p ≤ 0.05). The serum total oxidant status gradually increased as the tumour grade increased, being higher in grade four (11.96 ± 0.72) and lower in grade one (8.43 ± 1.56), and the mean differences were statistically significant (p ≤ 0 05). A statistically significantly higher total antioxidant capacity (116.78 ± 5.03 Trolox Eq/L) was obtained in the post-surgery than pre-surgery level (79.65 ± 17.914 Trolox Eq/L) (p ≤ 0 05). Higher oxidant and lower antioxidant levels were found in the serum of brain tumour patients than in the control group. The post-surgery oxidant level was lower than the pre-surgery state. The findings of this study could suggest that redox imbalance may have a role in the pathophysiology of brain tumours, but further experimental studies are needed.


Osmoregulation by Oat Coleoptile Protoplasts (Effect of Auxin).
The effect of auxin on the physiology of protoplasts from growing oat (Avena sativa L.) coleoptiles was investigated. Protoplasts, isolated iso-osmotically from peeled oat coleoptile segments, were found to swell steadily over many hours. Incubated in 1 mM CaCl2, 10 mM KCl, 10 mM 2-(morpholino)ethanesulfonic acid/1,3-bis-[tris(hydroxymethyl)methylamino]propane, pH 6.5, and mannitol to 300 milliosmolal, protoplasts swelled 28.9% [plus or minus] 2.0 (standard error) after 6 h. Addition of 10 [mu]M indoleacetic acid (IAA) increased swelling to 41.1% [plus or minus] 2.1 (standard error) after 6 h. Swelling (in the absence of IAA) was partially dependent on K+ in the bath medium, whereas auxin-induced swelling was entirely dependent on K+. Replacement of mannitol in the bath by Glc increased swelling (in the absence of IAA) and eliminated auxin-induced swelling. Swelling with or without IAA was inhibited by osmotic shock and was completely reversed by 0.1 mM NaN3. Sodium orthovanadate, applied at 0.5 mM, only gradually inhibited swelling under various conditions but was most effective with protoplasts prepared from tissue preincubated in vanadate. Our data are interpreted to suggest that IAA increases the conductance of the plasma membrane to K+.


Similarity relations in visual search predict rapid visual categorization.
How do we perform rapid visual categorization?It is widely thought that categorization involves evaluating the similarity of an object to other category items, but the underlying features and similarity relations remain unknown. Here, we hypothesized that categorization performance is based on perceived similarity relations between items within and outside the category. To this end, we measured the categorization performance of human subjects on three diverse visual categories (animals, vehicles, and tools) and across three hierarchical levels (superordinate, basic, and subordinate levels among animals). For the same subjects, we measured their perceived pair-wise similarities between objects using a visual search task. Regardless of category and hierarchical level, we found that the time taken to categorize an object could be predicted using its similarity to members within and outside its category. We were able to account for several classic categorization phenomena, such as (a) the longer times required to reject category membership; (b) the longer times to categorize atypical objects; and (c) differences in performance across tasks and across hierarchical levels. These categorization times were also accounted for by a model that extracts coarse structure from an image. The striking agreement observed between categorization and visual search suggests that these two disparate tasks depend on a shared coarse object representation.


Variations in the optical scattering properties of phytoplankton cultures.
The scattering and backscattering coefficients of 15 phytoplankton species were determined in the laboratory using the acs and BB9 instruments. The spectral variability of scattering properties was investigated and the homogenous sphere model based on Mie theory was also evaluated. The scattering efficiencies at 510 nm varied from 1.42 to 2.26, and the backscattering efficiencies varied from 0.003 to 0.020. The backscattering ratios at 510 nm varied from 0.17% to 0.97%, with a mean value of 0.58%. The scattering properties were influenced by algal cell size and cellular particulate organic carbon content rather than the chlorophyll a concentration. Comparison of the measured results to the values estimated using the homogenous sphere model showed that: (1) The model could well reproduce the spectral scattering coefficient with relative deviations of 5-39%, which indicates that cell shape and internal structure have no significant effects on predicting the scattering spectra; (2) Although the homogenous sphere model generally reflected the spectral trend of backscattering spectra for most species, it severely underestimated the backscattering coefficients by 1.4-48.6 folds at 510 nm. The deviations for Chaetoceros sp. and Microcystis aeruginosa were large and might be due to algal cell chain links and intracellular gas vacuoles, respectively.


Unusual topogenic sequence directs prion protein biogenesis.
Biosynthetic studies of the prion protein (PrP) have shown that two forms of different topology can be generated from the same pool of nascent chains in cell-free translation systems supplemented with microsomal membranes. A transmembrane form is the predominant product generated in wheat germ (WG) extracts, whereas a completely translocated (secretory) form is the major product synthesized in rabbit reticulocyte lysates (RRL). An unusual topogenic sequence within PrP is now shown to direct this system-dependent difference. The actions of this topogenic sequence were independent of on-going translation and could be conferred to heterologous proteins by the engineering of a discrete set of codons. System-dependent topology conferred by addition of RRL to WG translation products suggests that this sequence interacts with one or more cytosolic factors.


Deaths with transdermal fentanyl patches.
Fentanyl is a potent Schedule II narcotic analgesic recommended for use in the management of unremitting pain not controlled by morphine or other opiate/opioid drugs. The danger inherent to fentanyl is its potency (greater than 50-100 times that of morphine) and rapidity of action, causing respiratory depression within minutes of administration. Advisories have been issued on a state and national level to health care providers and through manufacturers' package inserts for patients. Still, as will be demonstrated in this case review, the use of only a single transdermal patch taken as prescribed for the first time can prove fatal. A drug that requires such extensive warnings-that if unheeded lead to death because of its narrow therapeutic/toxic window, should have strict criteria and limited outpatient use. Initial medical observation and documentation for determining tolerance might be required before issuing a prescription. There has been a rise in the popularity of this drug evidenced by increased deaths among drug abusers and more prescriptions written. In the year 2006, the Center for Forensic Sciences in Onondaga County had 8 cases where fentanyl was considered the cause of death, often with other drugs detected in therapeutic concentrations. This number was a marked increase from the 1 to 2 cases occurring annually from 2002 to 2005. All of these 2006 overdoses because of fentanyl involved the transdermal formulation. The investigative data, blood and liver fentanyl levels, and autopsy findings will be presented.


A conserved structural element in the RNA helicase UPF1 regulates its catalytic activity in an isoform-specific manner.
The RNA helicase UPF1 is a key component of the nonsense mediated mRNA decay (NMD) pathway. Previous X-ray crystal structures of UPF1 elucidated the molecular mechanisms of its catalytic activity and regulation. In this study, we examine features of the UPF1 core and identify a structural element that adopts different conformations in the various nucleotide- and RNA-bound states of UPF1. We demonstrate, using biochemical and single molecule assays, that this structural element modulates UPF1 catalytic activity and thereby refer to it as the regulatory loop. Interestingly, there are two alternatively spliced isoforms of UPF1 in mammals which differ only in the lengths of their regulatory loops. The loop in isoform 1 (UPF11) is 11 residues longer than that of isoform 2. We find that this small insertion in UPF11 leads to a two-fold increase in its translocation and ATPase activities. To determine the mechanistic basis of this differential catalytic activity, we have determined the X-ray crystal structure of the helicase core of UPF11 in its apo-state. Our results point toward a novel mechanism of regulation of RNA helicases, wherein alternative splicing leads to subtle structural rearrangements within the protein that are critical to modulate enzyme movements and catalytic activity.


Prevention and early detection of ovarian cancer: mission impossible?
Epithelial ovarian cancer is neither a common nor a rare disease. In the United States, the prevalence of ovarian cancer in postmenopausal women (1 in 2,500) significantly affects strategies for prevention and detection. If chemoprevention for ovarian cancer were provided to all women over the age of 50, side effects would have to be minimal in order to achieve an acceptable ratio of benefit to risk. This ratio might be improved by identifying subsets of individuals at increased risk or by bundling prevention of ovarian cancer with treatment for other more prevalent conditions. Approximately 10% of ovarian cancers are familial and relate to mutations of BRCA1, BRCA2, and mismatch repair genes. More subtle genetic factors are being sought in women with apparently sporadic disease. Use of oral contraceptive agents for as long as 5 years decreases the risk of ovarian cancer in later life by 50%. In one study, fenretinide (4-HPR) delayed development of ovarian cancer in women at increased risk of developing breast and ovarian cancer. Accrual to confirmatory studies has been prohibitively slow and prophylactic oophorectomy is recommended for women at increased genetic risk. Vaccines may have a role for prevention of several different cancers. Breast and ovarian cancers express mucins that could serve as targets for vaccines to prevent both cancers. Early detection of ovarian cancer requires a strategy with high sensitivity (> 75% for stage I disease) and very high specificity (> 99.6%) to achieve a positive predictive value of 10%. Transvaginal sonography (TVS) has achieved these values in some studies, but is limited by the cost of annual screening in a general population. Two-stage strategies that incorporate both serum markers and TVS promise to be more cost-effective. An algorithm has been developed that calculates risk of ovarian cancer based on serial CA125 values and refers patients at highest risks for TVS. Use of the algorithm is currently being evaluated in a trial with 200,000 women in the United Kingdom that will critically test the ability of a two-stage screening strategy to improve survival in ovarian cancer. Whatever the outcome, additional serum markers will be required to detect all patients in an initial phase of screening. More than 30 serum markers have been evaluated alone and in combination with CA125. Recent candidates include: HE4, mesothelin, M-CSF, osteopontin, kallikrein(s) and soluble EGF receptor. Proteomic approaches have been used to define a distinctive pattern of peaks on mass spectroscopy or to identify a limited number of critical markers that can be assayed by more conventional methods. Several groups are placing known markers on multiplex platforms to permit simultaneous assay of multiple markers with very small volumes of serum. Mathematical techniques are being developed to analyze combinations of marker levels to improve sensitivity and specificity. In the future, serum markers should improve the sensitivity of detecting recurrent disease as well as facilitate earlier detection of ovarian cancer.


Eye fixation during multiple object attention is based on a representation of discrete spatial foci.
We often look at and attend to several objects at once. How the brain determines where to point our eyes when we do this is poorly understood. Here we devised a novel paradigm to discriminate between different models of spatial selection guiding fixation. In contrast to standard static attentional tasks where the eye remains fixed at a predefined location, observers selected their own preferred fixation position while they tracked static targets that were arranged in specific geometric configurations and which changed identity over time. Fixations were best predicted by a representation of discrete spatial foci, not a polygonal grouping, simple 2-foci division of attention or a circular spotlight. Moreover, attentional performance was incompatible with serial selection. Together with previous studies, our findings are compatible with a view that attentional selection and fixation rely on shared spatial representations and suggest a more nuanced definition of overt vs. covert attention.


Protection against pathogen and salt stress by four plant growth-promoting rhizobacteria isolated from Pinus sp. on Arabidopsis thaliana.
The ability of four plant growth-promoting rhizobacteria, isolated in a previous study, to induce systemic resistance on Arabidopsis thaliana Col 0 against biotic and abiotic stress was evaluated. All the bacteria enhanced protection against the foliar pathogen Pseudomonas syringae DC3000 and increased plant tolerance to salt stress (NaCl 60 mM). Bacillus sp. strain L81 and Arthrobacter oxidans strain BB1 performed best with a decrease in the disease index of 61.2 and 52.3%, respectively, and a reduction in the mortality due to salt stress of 72.4 and 57.8%, respectively. Additionally, significant differences were found in growth and photosynthesis, again, L81 and BB1 performed best either in normal or under stress conditions. In order to elucidate the pathway elicited by these two strains to induce systemic resistance, experiments with the transgenic line of Arabidopsis thaliana NahG (defective in salicylic acid [SA]) and with the jar1 mutant (defective in jasmonic acid) were carried out. Results showed that the SA-dependent pathway was involved in the defense response induced by strains L81 and BB1. Results from quantitative reverse transcription-polymerase chain reaction analysis of the PR1 gene, related to the SA-dependent pathway and the PDF1.2 gene related to the SA-independent pathway, showed an increased expression of PR1 in BB1-treated plants, confirming involvement of the SA-dependent pathway in the defensive response.


Hypoxia induces an immunodominant target of tuberculosis specific T cells absent from common BCG vaccines.
M. tuberculosis (MTB) species-specific antigenic determinants of the human T cell response are important for immunodiagnosis and vaccination. As hypoxia is a stimulus in chronic tuberculosis infection, we analyzed transcriptional profiles of MTB subject to 168 hours of hypoxia to test the hypothesis that upregulation by hypoxia might result in gene products being recognized as antigens. We identified upregulation of two region of difference (RD) 11 (Rv2658C and Rv2659c), and one RD2 (Rv1986) absent from commonly used BCG strains. In MTB infected persons, the IL-2 ELISpot response to Rv1986 peptides was several times greater than the corresponding IFN-γ response to the reference immunodominant ESAT-6 or CFP-10 antigens. The IL-2 response was confined to two epitopic regions containing residues 61-80 and 161-180. The biggest population of IL-2 secreting T cells was single cytokine positive central memory T cells. The IL-2 response to live MTB bacilli lacking Rv1986 was significantly lower than the response to wild type or mutant complemented with Rv1986. In addition, the IL-2 response to Rv1986 was significantly lower in HIV-TB co-infected persons than in HIV uninfected persons, and significantly increased during antiretroviral therapy. These findings demonstrate that Rv1986 is an immunodominant target of memory T cells and is therefore of relevance when considering the partial efficacy of currently used BCG vaccines and provide evidence for a clinical trial comparing BCG strains.


Systematic review of psychosocial factors at work and private life as risk factors for back pain.
A systematic review of observational studies. To assess whether psychosocial factors at work and in private life are risk factors for the occurrence of back pain. Several reviews on risk factors for back pain have paid attention to psychosocial factors. However, in none of the published reviews was a strict systematic approach used to identify and summarize the available evidence. A computerized bibliographical search of several databases was performed, restricted to studies with a cohort or case-control design. A rating system was used to assess the strength of the evidence for various factors, based on the methodologic quality of the studies and the consistency of the findings. Eleven cohort and two case-control studies were included in this review. Strong evidence was found for low social support in the workplace and low job satisfaction as risk factors for back pain. Insufficient evidence was found for an effect of a high work pace, high qualitative demands, low job content, low job control, and psychosocial factors in private life. Evidence was found for an effect of low workplace social support and low job satisfaction. However, the result for workplace social support was sensitive to slight changes in the rating system, and the effect found for low job satisfaction may be a result of insufficient adjustment for psychosocial work characteristics and physical load at work. In addition, the combined evaluation of job content and job control, both aspects of decision latitude, led to strong evidence of a role for low job decision latitude. Thus, based on this review, there is evidence for an effect of work-related psychosocial factors, but the evidence for the role of specific factors has not been established yet.


Central neck dissection via the transoral approach.
There has been a strong impetus for the development of remote access approaches to the central neck. The primary motivation for this has been to alleviate the negative impact that some patients may perceive from a central neck scar. Numerous approaches have been described; however the only approach that provides midline access and equivalent visualization of the bilateral thyroid lobes and paratracheal basins is transoral neck surgery (TONS). TONS has been shown to be safe and effective in performing thyroidectomy, parathyroidectomy, and central neck dissection (CND) via both the endoscopic and robotic techniques. In contrast with other remote access techniques, it provides the surgeon with familiar views of the bilateral recurrent laryngeal nerves (RLN) at their insertion site in concert with equivalent access to both paratracheal basins, thus uniquely facilitating safe and comprehensive CND. Though feasible and safe, CND via TONS is not appropriate in all cases. CND via TONS should only be performed with concomitant transoral total thyroidectomy, either prophylactically if the surgeon routinely performs prophylactic CND, or therapeutically if there is newly found evidence of nodal metastasis in the central compartment at the time of surgery. We base these recommendations on both the recent American Head and Neck Society (AHNS) consensus statement for indications for transcervical CND and the baseline indications for TONS.


Positive involuntary autobiographical memories: you first have to live them.
Involuntary autobiographical memories (IAMs) are typically discussed in the context of negative memories such as trauma 'flashbacks'. However, IAMs occur frequently in everyday life and are predominantly positive. In spite of this, surprisingly little is known about how such positive IAMs arise. The trauma film paradigm is often used to generate negative IAMs. Recently an equivalent positive film was developed inducing positive IAMs (Davies, Malik, Pictet, Blackwell, & Holmes, 2012). The current study is the first to investigate which variables (emotional reaction to the film; recognition memory of the film; participant characteristics) would best predict the frequency of positive IAMs. Higher levels of positive mood change to the film were significantly associated with the number of positive IAMs recorded in the subsequent week. Results demonstrate the importance of positive emotional reaction at the time of an event for subsequent positive IAMs.


Magnetic resonance studies of brain function and neurochemistry.
In the short time since its introduction, magnetic resonance imaging (MRI) has rapidly evolved to become an indispensable tool for clinical diagnosis and biomedical research. Recently, this methodology has been successfully used for the acquisition of functional, physiological, and biochemical information in intact systems, particularly in the human body. The ability to map areas of altered neuronal activity in the brain, often referred to as functional magnetic resonance imaging (fMRI), is probably one of the most significant recent achievements that rely on this methodology. This development has permitted the examination of functional specialization in human and animal brains with unprecedented spatial resolution, as demonstrated by mapping at the level of orientation and ocular dominance columns in the visual cortex. These functional imaging studies are complemented by the ability to study neurochemistry using magnetic resonance spectroscopy, allowing the determination of metabolic processes that support neurotransmission and neurotransmission rates themselves.


Outcomes in vestibular ablative procedures.
To assess dizziness handicap and postural recovery in 17 patients undergoing a vestibular ablative procedure. Outcomes were compared between nine patients who underwent selective vestibular neurectomy and eight patients who underwent a transmastoid labyrinthectomy. Patients underwent ablative procedures at a tertiary care facility. Patients were tested with posturography (sensory organization test [SOT]) and the Dizziness Handicap Inventory (DHI) before and 4 to 5 weeks after their procedure. The mean change scores for each of the DHI subscales and the total DHI score showed significant improvement for the labyrinthectomy group. For the neurectomy group, only the emotional DHI subscale change score showed significant improvement. No difference was noted between preprocedure and postprocedure SOT scores for either group. The results indicated that the largest proportion of significant correlations existed between DHI and the somatosensory and vestibular subtests of platform posturography. In regression analyses, a model with age (p = 0.04) and vestibular score (p = 0.001) fitted the data well and explained 52.9% of the variance. Persons who were less than 69 years old were three times more likely to report persistent episodes of dizziness after the ablative procedure, and those who had neurectomies were 2.3 times more likely to report episodes of dizziness than those who underwent transmastoid labyrinthectomy. Dizziness handicap after an ablative procedure is influenced by the type of procedure (labyrinthectomy versus neurectomy), age, and preoperative vestibular score on the SOT. Preoperative vestibular SOT score is highly associated with the emotional, physical, and functional domains of perceived handicap and quality of life after the surgery.


Advanced techniques in abdominal surgery.
Almost every abdominal organ is now amenable to laparoscopic surgery. Laparoscopic appendicectomy is a routine procedure which also permits identification of other conditions initially confused with an inflamed appendix. However, assessment of appendiceal inflammation is more difficult. Almost all colonic procedures can be performed laparoscopically, at least partly, though resection for colonic cancer is still controversial. For simple patch repair of perforated duodenal ulcers laparoscopy is ideal, and inguinal groin hernia can be repaired satisfactorily with a patch of synthetic mesh. Many upper abdominal procedures, however, still take more time than the open operations. These techniques reduce postoperative pain and the incidence of wound infections and allow a much earlier return to normal activity compared with open surgery. They have also brought new disciplines: surgeons must learn different hand-eye coordination, meticulous haemostasis is needed to maintain picture quality, and delivery of specimens may be problematic. The widespread introduction of laparoscopic techniques has emphasised the need for adequate training (operations that were straight-forward open procedures may require considerable laparoscopic expertise) and has raised questions about trainee surgeons acquiring adequate experience of open procedures.


Vitamin D and adenosine triphosphatase dependent on divalent cations in rat intestinal mucosa.
Intestinal brush borders prepared from vitamin D-deficient rats demonstrate increased susceptibility in vitro to fragmentation by shear forces or to loss of microvillus enzymes on treatment with EDTA. These effects are relatively nonspecific and are also observed in normal rats starved for 48 h. They may underlie prior observations that purport to demonstrate a vitamin D-dependent increase in brush border Ca-dependent ATPase. In addition, however, vitamin D increases ATPase activity dependent on certain divalent cations, including Ca and Zn, in whole-particulate suspensions pelleted by high-speed centrifugation of mucosal homogenates. This action is independent of changes in other microvillus enzymes, i.e. disaccharidases, and tissue distribution and cation specificity studies support the hypothesis that the mucosal whole-particulate ATPase is related to transport of Ca, Zn, and possibly other divalent cations.


Social class gradients in health during adolescence.
To review existing data on social class gradients in adolescent health and to examine whether such gradients exist in new data concerning US adolescents. Review of relevant publications and unpublished data; regression analyses using adolescent self reported health status data to determine whether there are gradients by social class, using three classes categorised by adolescent reported parental work status and education. Adolescents of ages 11-17. Findings from the literature indicate the presence of social class gradients in some but not all aspects of adolescent health. Results from new data showed social class gradients in several domains of health and in profiles of health. The likelihood of being satisfied with one's health, of being more resilient (better family involvement, better problem solving, more physical activity, better home safety), having higher school achievement, and of being in the best health profiles were significantly and progressively greater as social class rose. Moreover, the probability of being in the poorest health profile type group was progressively higher as social class declined. The review of existing data and the new findings support the existence of social class gradients in satisfaction with one's health, in resilience to health threats, in school achievement, and in being in the best health overall (as manifested by the health profiles composed of four major domains of health). The study had two especially notable findings: (1) the paucity of studies using the same or similar indicators, and (2) the consistent existence of social class gradients in characteristics related to subsequent health, particularly intake of nutritional foods and physical activity. The sparseness of existing data and the different aspects of health investigated in the relatively few studies underscore the need for (1) the development of conceptual models specifically focused on adolescent health and social class; (2) additional inquiry into the measurement of social class and adolescent perceptions of class; (3) inclusion of contextual variables in study design; and (4) longitudinal cohort studies to better understand the specific determinants of health during adolescence.


Does a myocardial infarction boost your (B cell) memory?
Graphical abstract Establishment of an autoreactive B cell memory after myocardial infarction: a working hypothesis. The demise of cardiac cells leads to the release of cryptoantigens that induce a humoral immune response that leads to accumulation of immunoglobulins in plaques and eventually amplifies atherogenesis at remote sites.


AAEM minimonograph #47: normative data in electrodiagnostic medicine. ff.
This article reviews, without mathematics, the important principles governing the acquisition and use of normative data in electrodiagnostic medicine. Common flaws in neurophysiological normative data include vague clinical criteria for establishing freedom from disease, samples that are too small and inadequately stratified, and application of Gaussian statistics to non-Gaussian variables. Other problematic issues concern the trade-off between permissible false-positivity and false-negativity in defining the limits of normative from sample data, test-retest variability, and the use of multiple independent test measurements in each electrodiagnostic examination. The following standards for normative data are proposed: (1) standardized objective determination of freedom from disease; (2) appropriately large sample of normal subjects; (3) proportional statification of normal subjects for known relevant variables; (4) test of Gaussian fit for application of Gaussian statistics; and (5) data presentation by percentiles when Gaussian fit is in doubt. Many existing normative studies in clinical neurophysiology do not meet these standards. High-quality normative data, readily accessible, is essential for the accurate electrodiagnosis of neuromuscular diseases.


Thyroid transcription factor 1: a marker for lung adenoarinoma in body cavity fluids.
Adenocarcinomas are the most common epithelial malignancies in body cavity fluids. Subclassification of adenocarcinomas according to primary site can be a challenging task. Thyroid transcription factor 1 (TTF-1) is a nuclear transcription factor that is expressed in normal lung, in thyroid, and in their neoplasms. Because thyroid carcinomas rarely metastasize to the serosal surfaces, the authors used TTF-1 as a marker to distinguish adenocarcinomas of the lung from carcinomas of other organs. The authors studied 113 body cavity fluids (92 pleural fluid samples and 21 ascitic samples) from 113 patients with a diagnosis of adenocarcinoma on the basis of routine cytology. The primary sites of origin were confirmed clinically or histologically for all patients. There were 39 adenocarcinomas of the lung, 24 adenocarcinomas of the breast, 34 adenocarcinomas of the genitourinary tract, and 16 adenocarcinomas of the gastrointestinal tract. Archival Papanicolaou-stained, cytocentrifuged slides were used without destaining for immunocytochemistry with the monoclonal TTF-1 antibody using a commercial available method. TTF-1 was expressed in 21 of 39 lung adenocarcinomas (54%). Intense nuclear staining was present in tumor cells that occurred in groups or in isolated form. In contrast, none of the other types of adenocarcinomas expressed TTF-1. TTF-1 is a highly specific marker for adenocarcinomas of the lung in body cavity fluids. Immunocytochemistry using this antibody can be performed easily on archival Papanicolaou-stained, cytocentrifuged slides of fluid specimens.


Multisite Phosphorylation Modulates the T Cell Receptor ζ-Chain Potency but not the Switchlike Response.
Multisite phosphorylation is ubiquitous in cellular signaling and is thought to provide signaling proteins with additional regulatory mechanisms. Indeed, mathematical models have revealed a large number of mechanisms by which multisite phosphorylation can produce switchlike responses. The T cell antigen receptor (TCR) is a multisubunit receptor on the surface of T cells that is a prototypical multisite substrate as it contains 20 sites that are distributed on 10 conserved immunoreceptor tyrosine-based activation motifs (ITAMs). The TCR ζ-chain is a homodimer subunit that contains six ITAMs (12 sites) and exhibits a number of properties that are predicted to be sufficient for a switchlike response. We have used cellular reconstitution to systematically study multisite phosphorylation of the TCR ζ-chain. We find that multisite phosphorylation proceeds by a nonsequential random mechanism, and find no evidence that multiple ITAMs modulate a switchlike response but do find that they alter receptor potency and maximum phosphorylation. Modulation of receptor potency can be explained by a reduction in molecular entropy of the disordered ζ-chain upon phosphorylation. We further find that the tyrosine kinase ZAP-70 increases receptor potency but does not modulate the switchlike response. In contrast to other multisite proteins, where phosphorylations act in strong concert to modulate protein function, we suggest that the multiple ITAMs on the TCR function mainly to amplify subsequent signaling.


Female genital tract malignancies in Uyo, South-South Nigeria.
Female genital tract malignancies cause a significant number of deaths in the developing world. These deaths are however usually overshadowed by maternal deaths from obstetric causes. A retrospective study of all histologically confirmed female genital tract malignancies seen in the histopathology department of the University of Uyo Teaching Hospital between January 2000 and December 2004. Sixty five female genital tract malignancies were studied. Majority of patients were in the fifth to seventh decades of life. Carcinoma of the cervix was the commonest cancer seen (49.2%) followed by ovarian cancer (21.5%). Carcinoma of the vulva was the least common cancer seen (1.5%). Squamous cell carcinoma was the commonest histological variety. Female genital tract malignancies are major causes of morbidity and mortality in Nigeria. There is thus a need for public enlightenment to increase the awareness of risk factors and the provision of regular cervical cancer screening services which should be free to encourage utilisation.


Immune response does not prevent homologous Porcine epidemic diarrhoea virus reinfection five months after the initial challenge.
The aim of the present study was to evaluate the duration of protective immunity against Porcine epidemic diarrhoea virus (PEDV). To do so, a two phases study was performed. In the first phase, 75 four-week-old pigs (group A) were orally inoculated (0 days post-inoculation; dpi) with a European PEDV G1b strain and 14 were kept as controls (group B). The second phase started five months later (154 dpi), when animals in group A were homologous challenged and animals in group B were challenged for first time. Clinical signs, viral shedding and immune responses were evaluated after each inoculation, including the determination of antibodies (ELISA and viral neutralization test, IgA and IgG ELISPOTs using peripheral blood mononuclear cells and lymph node cells) and the frequency of interferon-gamma (IFN-γ) secreting cells. During the first phase, loose stools/liquid faeces were observed in all group A animals. Faecal shedding of PEDV occurred mostly during the first 14 days but, in some animals, persisted until 42 dpi. All inoculated animals seroconverted for specific-PEDV IgG and IgA, and for neutralizing antibodies (NA). At 154 dpi, 77% of pigs were still positive for NA. After that, the homologous challenge resulted in a booster for IgG, IgA, NA, as well as specific-PEDV IgG, IgA and IFN-γ secreting cells. In spite of that, PEDV was detected in faeces of all pigs from group A, indicating that the immune response did not prevent reinfection, although the duration of the viral shedding and the total load of virus shed were significantly lower for previously challenged pigs (p < .05). Taken together, the results indicated that, potentially, maintenance of PEDV infection within an endemic farm may occur by transmission to and from previously infected animals and also indicates that sterilizing immunity is shorter than the productive life of pigs.


A Comparison of 4D DSA with 2D and 3D DSA in the Analysis of Normal Vascular Structures in a Canine Model.
4D DSA allows viewing of 3D DSA as a series of time-resolved volumes of a contrast bolus. There is no comparison of the accuracy of the anatomic information provided by 4D DSA with that available from conventional 2D and 3D DSA. Our purpose was to make this comparison by using a canine model. 2D, 3D, and 4D DSA acquisitions were performed in 5 canines from 3 catheter positions in the common carotid artery yielding 15 2D, 15 3D, and 15 4D datasets. For each territory, 3 vascular segments were chosen for comparison. Images were reviewed by 2 experienced neuroradiologists and were graded by the ability to visualize a segment, its filling direction, and preferred technique. Two visualization modes for 4D DSA were compared (volume-rendering technique and MIP). 4D DSA was preferred in 73.9% of the image sets; 2D, in 22.7%; and 3D, in 3.4%. 4D DSA MIP rendering yielded superior visualization of very small vessel details; the 4D DSA volume-rendering technique offered superior depth and overlap information and better visualization of the surface details of the vasculature. In this study, 4D DSA was preferred over 2D and 3D DSA for analysis of normal vasculature. The ability to provide any view of a vascular territory at any time during passage of a contrast bolus seems likely to reduce the need for many 2D acquisitions during diagnostic and therapeutic procedures. This then potentially translates into a reduction in radiation and contrast dose.


Modified femtosecond pulse shaper using microlens arrays.
We discuss a modified femtosecond pulse shaper that uses microlens arrays to convert the continuous band of frequencies normally obtained in the mask plane of a pulse shaper into a series of discrete spots. Our experiments demonstrate that this modification can improve pulse-shaping quality when modulator arrays with large interpixel gaps are used for Fourier-plane filtering.


Mechanisms of reactivation of latent tuberculosis infection due to SIV coinfection.
HIV is a major driver of tuberculosis (TB) reactivation. Depletion of CD4+ T cells is assumed to be the basis behind TB reactivation in individuals with latent tuberculosis infection (LTBI) coinfected with HIV. Nonhuman primates (NHPs) coinfected with a mutant simian immunodeficiency virus (SIVΔGY) that does not cause depletion of tissue CD4+ T cells during infection failed to reactivate TB. To investigate the contribution of CD4+ T cell depletion relative to other mechanisms of SIV-induced reactivation of LTBI, we used CD4R1 antibody to deplete CD4+ T cells in animals with LTBI without lentiviral infection. The mere depletion of CD4+ T cells during LTBI was insufficient in generating reactivation of LTBI. Instead, direct cytopathic effects of SIV resulting in chronic immune activation, along with the altered effector T cell phenotypes and dysregulated T cell homeostasis, were likely mediators of reactivation of LTBI. These results revealed important implications for TB control in HIV-coinfected individuals.


The relationship between temporal phase discrimination ability and the frequency doubling illusion.
The frequency doubling illusion (FDI) occurs when a low spatial frequency sinusoidal grating is modulated at high temporal frequencies--its apparent spatial frequency increases. A recent study suggests that this illusion is perceived due to a frequency-dependent loss of temporal phase encoding ability. We sought to elucidate the relationship between temporal phase encoding and the FDI by exploring the spatiotemporal characteristics of temporal phase discrimination (TPD) thresholds using a novel stimulus comprising three grating patches presented simultaneously in a triangular pattern. A reference grating was presented superiorly, and six degrees below two gratings (one a copy of the reference) were each randomly presented in one of two fixed positions. The odd grating had abutting regions of spatial half-cycles with alternate half-cycles locked in temporal phase. The temporal phase difference between adjoining half-cycles was varied between 0 degrees and 180 degrees via QUEST staircase--subjects had to identify which lower stimulus appeared different from the reference grating. TPD thresholds were measured for 0.25, 0.50, and 2.20 cpd stimulus at six temporal frequencies (1 to 28 Hz) at 2x, 4x, and 8x orientation identification contrast thresholds. For all subjects, thresholds were variable at low contrasts. At higher contrasts, TPD thresholds increase for 0.25 and 0.50 cpd gratings with increasing flicker rate. These data support the idea that frequency-dependent loss of temporal phase encoding ability could possibly underlie the FDI.


PKC isoforms interact with and phosphorylate DNMT1.
DNA methyltransferase 1 (DNMT1) has been shown to be phosphorylated on multiple serine and threonine residues, based on cell type and physiological conditions. Although recent studies have suggested that protein kinase C (PKC) may be involved, the individual contribution of PKC isoforms in their ability to phosphorylate DNMT1 remains unknown. The PKC family consists of at least 12 isoforms that possess distinct differences in structure, substrate requirement, expression and localization. Here we show that PKCα, βI, βII, δ, γ, η, ζ and μ preferentially phosphorylate the N-terminal domain of human DNMT1. No such phosphorylation of DNMT1 was observed with PKCε. Using PKCζ as a prototype model, we also found that PKC physically interacts with and phosphorylates DNMT1. In vitro phosphorylation assays conducted with recombinant fragments of DNMT1 showed that PKCζ preferentially phosphorylated the N-terminal region of DNMT1. The interaction of PKCζ with DNMT1 was confirmed by GST pull-down and co-immunoprecipitation experiments. Co-localization experiments by fluorescent microscopy further showed that endogenous PKCζ and DNMT1 were present in the same molecular complex. Endogenous PKCζ activity was also detected when DNMT1 was immunoprecipitated from HEK-293 cells. Overexpression of both PKCζ and DNMT1 in HEK-293 cells, but not of either alone, reduced the methylation status of genes distributed across the genome. Moreover, in vitro phosphorylation of DNMT1 by PKCζ reduced its methytransferase activity. Our results indicate that phosphorylation of human DNMT1 by PKC is isoform-specific and provides the first evidence of cooperation between PKCζ and DNMT1 in the control of the DNA methylation patterns of the genome.


A compilation of soybean ESTs: generation and analysis.
Whole-genome sequencing is fundamental to understanding the genetic composition of an organism. Given the size and complexity of the soybean genome, an alternative approach is targeted random-gene sequencing, which provides an immediate and productive method of gene discovery. In this study, more than 120000 soybean expressed sequence tags (ESTs) generated from more than 50 cDNA libraries were evaluated. These ESTs coalesced into 16928 contigs and 17336 singletons. On average, each contig was composed of 6 ESTs and spanned 788 bases. The average sequence length submitted to dbEST was 414 bases. Using only those libraries generating more than 800 ESTs each and only those contigs with 10 or more ESTs each, correlated patterns of gene expression among libraries and genes were discerned. Two-dimensional qualitative representations of contig and library similarities were generated based on expression profiles. Genes with similar expression patterns and, potentially, similar functions were identified. These studies provide a rich source of publicly available gene sequences as well as valuable insight into the structure, function, and evolution of a model crop legume genome.


Iodine from brown algae in human nutrition, with an emphasis on bioaccessibility, bioavailability, chemistry, and effects of processing: A systematic review.
Brown algae are becoming increasingly popular as a food source and dietary supplement in Europe and other Western countries. As they are highly rich in iodine, they represent a potential new dietary iodine source. Iodine deficiency has been re-emerging in Europe, and it is important to ensure adequate intake through one's diet. However, macroalgae, and especially brown algae, may contain very high amounts of iodine, and both iodine deficiency and excessive iodine may increase the risk of negative health effects. The iodine content of algae or foods containing algae is currently not regulated in the European Union. The aim of this paper is to review the literature to determine the chemical species of iodine in brown algae, the loss of iodine during processing, and the bioavailability and bioaccessibility of iodine. A systematic search of the literature was performed in April 2021, via the databases Web of Science and PubMed. The review includes studies of iodine in brown macroalgae in relation to bioavailability, bioaccessibility, processing and speciation. A meta-analysis was conducted in relation to the following topics: (i) the correlation between total iodine and iodide (I- ) content in brown algae; (ii) the correlation between the loss of iodine during processing and the I- content; and (iii) the correlation between bioavailability and the I- content. The bioavailability of iodine from brown algae was generally high, with in vivo bioavailability ranging from 31% to 90%. The in vitro bioavailability of iodine (2%-28%) was systematically lower than in vivo bioavailability (31%-90%), indicating an inadequate in vitro methodology. Processing may reduce the iodine content of brown algae, and a higher I- content was positively correlated with increased iodine loss during processing. Although processing strategies may reduce the iodine content of brown algae significantly, the iodine content may still be high after processing. These findings may be used in food safety evaluations of brown algae as well as in the development of macroalgae-containing foods with iodine contents suitable for human consumption. Further research on processing techniques to reduce the iodine content in brown macroalgae are warranted.


Uterine fibroids: a review of health-related quality of life assessment.
To summarize the literature describing the impact of uterine fibroids on health-related quality of life (HRQOL), the measures used to assess HRQOL, and related areas for future research. A search of three electronic databases (MEDLINE, Embase, and Current Contents) was performed, and 40 papers were identified referencing leiomyoma or uterine fibroids and (health-related) quality of life (HRQOL). Studies including women with benign gynecological conditions other than uterine fibroids were also reviewed. The EQ-5D, SF-36, and related generic measures are used to assess HRQOL in women with diverse gynecological conditions, including uterine fibroids. The UFS-QOL, a condition- specific measure, provides scores on symptom severity and six HRQOL dimensions. All instruments consistently demonstrate that HRQOL is considerably impaired in women with symptomatic uterine fibroids and that appropriate treatment can lead to improved QOL. The published data on HRQOL associated with uterine fibroids report significantly lower HRQOL scores for women with fibroids than for women without this disorder. Additional research is needed to confirm these findings and provide greater detail describing the specific domains most affected by uterine fibroids. These advances are necessary to effectively compare new treatment modalities vs. standard invasive therapies, such as hysterectomy.


Prime-Boost Vaccine Regimen for SjTPI and SjC23 Schistosome Vaccines, Increases Efficacy in Water Buffalo in a Field Trial in China.
Schistosomiasis remains a serious zoonotic disease in China and the Philippines. Water buffalo and cattle account for the majority of transmission. Vaccination of water buffalo is considered a key strategy to reduce disease prevalence. Previously, we showed that vaccination of water buffalo with SjC23 or SjCTPI plasmid DNA vaccines, induced 50% efficacy to challenge infection. Here, we evaluated several parameters to determine if we can develop a two dose vaccine that maintains the efficacy of the three dose vaccine. We performed four trials evaluating: (1) lab produced vs. GLP grade vaccines, (2) varying the time between prime and boost, (3) the influence of an IL-12 adjuvant, and (4) a two dose heterologous (DNA-protein) prime-boost. We found the source of the DNA vaccines did not matter, nor did increasing the interval between prime and boost. Elimination of the IL-12 plasmid lowered homologous DNA-DNA vaccine efficacy. A major finding was that the heterologous prime boost improved vaccine efficacy, with the prime-boost regimen incorporating both antigens providing a 55% reduction in adult worms and 53% reduction in liver eggs. Vaccinated buffalo produced vaccine-specific antibody responses. These trials suggest that highly effective vaccination against schistosomes can be achieved using a two dose regimen. No adjuvants were used with the protein boost, and the potential that addition of adjuvant to the protein boost to further increase efficacy should be evaluated. These results suggest that use of these two schistosome vaccines can be part of an integrated control strategy to reduce transmission of schistosomiasis in Asia.


A mathematical model for suppression subtractive hybridization.
Suppression subtractive hybridization (SSH) is frequently used to unearth differentially expressed genes on a whole-genome scale. Its versatility is based on combining cDNA library subtraction and normalization, which allows the isolation of sequences of varying degrees of abundance and differential expression. SSH is a complex process with many adjustable parameters that affect the outcome of gene isolation.We present a mathematical model of SSH based on DNA hybridization kinetics for assessing the effect of various parameters to facilitate its optimization. We derive an equation for the probability that a particular differentially expressed species is successfully isolated and use this to quantify the effect of the following parameters related to the cDNA sample: (a) mRNA abundance; (b) partial sequence complementarity to other species; and (3) degree of differential expression. We also evaluate the effect of parameters related to the process, including: (a) reaction times; and (b) extent of driver excess used in the two hybridization reactions. The optimum set of process parameters for successful isolation of differentially expressed species depends on transcript abundance. We show that the reaction conditions have a significant effect on the occurrence of false-positives and formulate strategies to isolate specific subsets of differentially expressed genes. We also quantify the effect of non-specific hybridization on the false-positive results and present strategies for spiking cDNA sequences to address this problem.


Some comments and suggestions concerning population pharmacokinetic modeling, especially of digoxin, and its relation to clinical therapy.
Population pharmacokinetic and dynamic modeling is often employed to analyze data of steady-state trough serum digoxin concentrations in the course of what is frequently regarded as routine therapeutic drug monitoring (TDM). Such a monitoring protocol is extremely uninformative. It permits only the estimation of a single parameter of a 1-compartment model, such as clearance. The use of D-optimal design strategies permits much more information to be obtained, employing models having a really meaningful structure. Strategies and protocols for routine TDM policies greatly need to be improved, incorporating these principles of optimal design. Software for population pharmacokinetic modeling has been dominated by NONMEM. However, because NONMEM is a parametric method, it must assume a shape for the model parameter distributions. If the assumption is not correct, the model will be in error, and the most likely results given the raw data will not be obtained. In addition, the likelihood as computed by NONMEM is only approximate, not exact. This impairs statistical consistency and reduces statistical efficiency and the resulting precision of model parameter estimates. Other parametric methods are superior, as they provide exact likelihoods. However, they still suffer from the constraints of assuming the shape of the model parameter distributions. Nonparametric methods are more flexible. One need not make any assumptions about the shape of the parameter distributions. Nonparametric methods also provide exact likelihoods and are statistically consistent, efficient, and precise. They also permit maximally precise dosage regimens to be developed for patients using multiple model dosage design, something parametric modeling methods cannot do. Laboratory assay errors are better described by the reciprocal of the assay variance of each measurement rather than by coefficient of variation. This is easy to do and permits more precise models to be made. This also permits estimation of assay error separately from the other sources of uncertainty in the clinical environment. This is most useful scientifically. Digoxin has at least 2-compartment behavior. Its pharmacologic and clinical effects correlate not with serum digoxin concentrations but with those in the peripheral nonserum compartment. Some illustrative clinical examples are discussed. It seems that digitalis therapy, guided by TDM and our 2 compartment models based on that of Reuning et al, can convert at least some patients with atrial fibrillation and flutter to regular sinus rhythm. Investigators have often used steady-state trough concentrations only to make a 1-compartment model and have sought only to predict future steady-state trough concentrations. Much more than this can be done, and clinical care can be much improved. Further work along these lines is greatly to be desired.


PML/RARalpha plays a role for basal activity and retinoid-induced repression of the tissue factor promoter in acute promyelocytic leukemia cells.
Constitutive expression of tissue factor (TF) by acute promyelocytic leukemia (APL) cells may contribute to thrombotic complications. In this study we examined the transcriptional mechanisms of all-trans retinoic acid (ATRA)-induced down-regulation of TF in the APL cell line NB4, by analysis of stable clones expressing the luciferase gene under the control of 5' flanking regions of the TF gene. We show that the TF promoter is constitutively active in NB4 cells, and that ATRA induces rapid suppression of the promoter. Basal activity and ATRA-induced suppression of TF promoter is determined by the proximal -383 to +121 bp of the promoter. Electrophoretic mobility shift assays demonstrate the binding of Fos/Jun complexes to two TF promoter AP-1 sites in this region. Both complexes were suppressed by ATRA treatment. The ectopic expression of the APL-specific PML/RARalpha oncoprotein in U-937 cells results in induction of TF mRNA and promoter activity. Interestingly, this PML/RARalpha-mediated increase in TF promoter activity is sensitive to ATRA treatment. These data indicate that TF expression in APL cells is exacerbated by the presence of the PML/RARalpha fusion protein, and implicates the loss of Fos/Jun binding to the TF promoter in ATRA-induced suppression of TF.


Prevertebrate Local Gene Duplication Facilitated Expansion of the Neuropeptide GPCR Superfamily.
In humans, numerous genes encode neuropeptides that comprise a superfamily of more than 70 genes in approximately 30 families and act mainly through rhodopsin-like G protein-coupled receptors (GPCRs). Two rounds of whole-genome duplication (2R WGD) during early vertebrate evolution greatly contributed to proliferation within gene families; however, the mechanisms underlying the initial emergence and diversification of these gene families before 2R WGD are largely unknown. In this study, we analyzed 25 vertebrate rhodopsin-like neuropeptide GPCR families and their cognate peptides using phylogeny, synteny, and localization of these genes on reconstructed vertebrate ancestral chromosomes (VACs). Based on phylogeny, these GPCR families can be divided into five distinct clades, and members of each clade tend to be located on the same VACs. Similarly, their neuropeptide gene families also tend to reside on distinct VACs. Comparison of these GPCR genes with those of invertebrates including Drosophila melanogaster, Caenorhabditis elegans, Branchiostoma floridae, and Ciona intestinalis indicates that these GPCR families emerged through tandem local duplication during metazoan evolution prior to 2R WGD. Our study describes a presumptive evolutionary mechanism and development pathway of the vertebrate rhodopsin-like GPCR and cognate neuropeptide families from the urbilaterian ancestor to modern vertebrates.


What leads some people to think they are HIV-positive before knowing their diagnosis? A systematic review of psychological and behavioural correlates of HIV-risk perception.
Current HIV-risk perception refers to the extent to which individuals think they might be HIV-positive. This belief, distinct from the perceived risk about being infected with HIV in the future, is likely to have a range of important consequences. These consequences may include both psychological effects (e.g., impacts on well-being) and behavioural effects (e.g., HIV testing uptake). Given these possible outcomes, and the suggested importance of risk perception in health behaviour models, understanding the behavioural and psychological antecedents of current HIV-risk perception is crucial. This systematic review investigates the relationship between behavioural and psychological factors and current HIV-risk perception (in individuals who are unaware of their actual HIV status). Eight studies were eligible for inclusion in the review (five quantitative and three qualitative studies). Drug risk behaviour and sexual risk behaviour (both self and partner) were often associated with current HIV-risk perception, although other studies failed to show a relationship between one's own sexual risk behaviour and risk perception. Psychological factors were only rarely assessed in relation to current HIV-risk perception. Where these variables were included, there was evidence that experiencing symptoms perceived to be consistent with HIV and prompts to test were associated with increased current HIV-risk perception. These findings are consistent with the Common-Sense Model (CSM) of illness representation and self-regulation. Methodological quality criteria were rarely met for the included studies. In addition, it was often difficult to ascertain whether potentially includable studies were eligible due to imprecise definitions of HIV-risk perception. Research and practice implications are discussed, with particular emphasis on the role of risk appraisals as a potential mediator of the relationship between HIV-risk behaviour, symptoms and current HIV-risk perception.


Inflammation and lung injury in an ovine model of fluid resuscitated endotoxemic shock.
Sepsis is a multi-system syndrome that remains the leading cause of mortality and critical illness worldwide, with hemodynamic support being one of the cornerstones of the acute management of sepsis. We used an ovine model of endotoxemic shock to determine if 0.9% saline resuscitation contributes to lung inflammation and injury in acute respiratory distress syndrome, which is a common complication of sepsis, and investigated the potential role of matrix metalloproteinases in this process. Endotoxemic shock was induced in sheep by administration of an escalating dose of lipopolysaccharide, after which they subsequently received either no fluid bolus resuscitation or a 0.9% saline bolus. Lung tissue, bronchoalveolar fluid (BAL) and plasma were analysed by real-time PCR, ELISA, flow cytometry and immunohistochemical staining to assess inflammatory cells, cytokines, hyaluronan and matrix metalloproteinases. Endotoxemia was associated with decreased serum albumin and total protein levels, with activated neutrophils, while the glycocalyx glycosaminoglycan hyaluronan was significantly increased in BAL. Quantitative real-time PCR studies showed higher expression of IL-6 and IL-8 with saline resuscitation but no difference in matrix metalloproteinase expression. BAL and tissue homogenate levels of IL-6, IL-8 and IL-1β were elevated. This data shows that the inflammatory response is enhanced when a host with endotoxemia is resuscitated with saline, with a comparatively higher release of inflammatory cytokines and endothelial/glycocalyx damage, but no change in matrix metalloproteinase levels.


Carbon monoxide binding by de novo heme proteins derived from designed combinatorial libraries.
Carbon monoxide binding was studied in a collection of de novo heme proteins derived from combinatorial libraries of sequences designed to fold into 4-helix bundles. The design of the de novo sequences was based on the previously reported "binary code" strategy, in which the patterning of polar and nonpolar amino acids is specified explicitly, but the exact identities of the side chains are varied extensively.(1) The combinatorial mixture of amino acids included histidine and methionine, which ligate heme iron in natural proteins. However, no attempt was made to explicitly design a heme binding site. Nonetheless, as reported previously, approximately half of the binary code proteins bind heme.(2) This collection of novel heme proteins provides a unique opportunity for an unbiased assessment of the functional potentialities of heme proteins that have not been prejudiced either by explicit design or by evolutionary selection. To assess the capabilities of the de novo heme proteins to bind diatomic ligands, we measured the affinity for CO, the kinetics of CO binding and release, and the resonance Raman spectra of the CO complexes for eight de novo heme proteins from two combinatorial libraries. The CO binding affinities for all eight proteins were similar to that of myoglobin, with dissociation constants (K(d)) in the low nanomolar range. The CO association kinetics (k(on)) revealed that the heme environment in all eight of the de novo proteins is partially buried, and the resonance Raman studies indicated that the local environment around the bound CO is devoid of hydrogen-bonding groups. Overall, the CO binding properties of the de novo heme proteins span a narrow range of values near the center of the range observed for diverse families of natural heme proteins. The measured properties of the de novo heme proteins can be considered as a "default" range for CO binding in alpha-helical proteins that have neither been designed to bind heme or CO, nor subjected to genetic selections for heme or CO binding.


Active control of the volume acquisition noise in functional magnetic resonance imaging: method and psychoacoustical evaluation.
Functional magnetic resonance imaging (fMRI) provides a noninvasive tool for observing correlates of neural activity in the brain while a subject listens to sound. However, intense acoustic noise is generated in the process of capturing MR images. This noise stimulates the auditory nervous system, limiting the dynamic range available for displaying stimulus-driven activity. The noise is potentially damaging to hearing and is distracting for the subject. In an active noise control (ANC) system, a reference sample of a noise is processed to form a sound which adds destructively with the noise at the listener's ear. We describe an implementation of ANC in the electromagnetically hostile and physically compact MRI scanning environment. First, a prototype system was evaluated psychoacoustically in the laboratory, using the electrical drive to a noise-generating loudspeaker as the reference. This system produced 10-20 dB of subjective noise-reduction between 250 Hz and 1 kHz, and smaller amounts at higher frequencies. The system was modified to operate in a real MR scanner where the reference was obtained by recording the acoustic scanner noise. Objective reduction by 30-40 dB of the most intense component in scanner noises was realized between 500 Hz and 3500 Hz, and subjective reduction of 12 dB and 5 dB in tests at frequencies of 600 Hz and at 1.9 kHz, respectively. Although the benefit of ANC is limited by transmission paths to the cochlea other than air-conduction routes from the auditory meatus, ANC achieves worthwhile attenuation even in the frequency range of maximum bone conduction (1.5-2 kHz). ANC should, therefore, be generally useful during auditory fMRI.