Biphasic recombination of photodissociated CO compound of cytochrome o(s) from Vitreoscilla.
The soluble cytochrome o from Vitreoscilla contains two identical subunits and two hemes. The reduced form binds 2 mol of CO in a cooperative manner with a Hill coefficient near 2 (Tyree, B., and Webster, D. A. (1978) J. Biol. Chem. 253, 6988-6991). This carbonyl compound was photolysed with a dye laser and recombination followed at 437 or 420 nm where maximal absorbance changes were registered. Recombination kinetics were biphasic, and the fast phase was approximately 10 times the rate of the slow phase. Apparent rate constants of both phases showed a nonlinear dependence on CO concentration, respectively, in conformity with a reaction scheme which assumes the transient formation of an intermediate species in both slow and fast reactions. A study of temperature dependence of the reactions gave EA = 2.7 kcal/mol for the slow reaction and EA = 3.2 kcal/mol for the fast reaction below 23 degrees C; above this temperature the slope of the Arrhenius plot for the fast reaction became positive. Maximal rates for both phases were around pH 6.5 and fell to approximately 40% of maximal at pH 12. The binding reaction was affected by even a low concentration of sodium dodecyl sulfate (0.0025%), which changed both the kinetic constant of each phase and the relative contribution of each phase to the reaction. A model which assumes the existence of fast and slow reaction conformers in equilibrium is proposed.


Fathers' Needs and Masculinity Dilemmas in a Neonatal Intensive Care Unit in Denmark.
Most healthcare professionals in neonatal intensive care units typically focus on the infants and mothers; fathers often feel powerless and find it difficult to establish a father-child relationship. In family-centered healthcare settings, exploring fathers' experiences and needs is important because men's roles in society, especially as fathers, are changing. To describe fathers' needs when their infants are admitted to a neonatal intensive care unit and to discuss these needs within a theoretical framework of masculinity to advance understanding and generate meaningful knowledge for clinical practices. This qualitative study used participant observation, interviews, multiple sequential interviews, and a focus group discussion. Data were analyzed using grounded theory principles. Analysis of the fathers' needs generated 2 primary themes: (1) Fathers as caregivers and breadwinners and (2) fathers and emotions. Fathers wished to be involved and to take care of their infants but have to balance cultural and social norms and expectations of being breadwinners with their wishes to be equal coparents. Health professionals in neonatal intensive care units must be aware of fathers' need and desire to be equal coparents. Nurses should play a key role by, for example, showing that fathers are as important to their infants as are the mothers, helping them become involved in childcare, and ensuring that they are directly informed about their children's progress. Further research in other cultural settings would contribute to knowledge regarding fatherhood and the role of fathers in childcare.


The Arabidopsis halophytic relative Thellungiella halophila tolerates nitrogen-limiting conditions by maintaining growth, nitrogen uptake, and assimilation.
A comprehensive knowledge of mechanisms regulating nitrogen (N) use efficiency is required to reduce excessive input of N fertilizers while maintaining acceptable crop yields under limited N supply. Studying plant species that are naturally adapted to low N conditions could facilitate the identification of novel regulatory genes conferring better N use efficiency. Here, we show that Thellungiella halophila, a halophytic relative of Arabidopsis (Arabidopsis thaliana), grows better than Arabidopsis under moderate (1 mm nitrate) and severe (0.4 mm nitrate) N-limiting conditions. Thellungiella exhibited a lower carbon to N ratio than Arabidopsis under N limitation, which was due to Thellungiella plants possessing higher N content, total amino acids, total soluble protein, and lower starch content compared with Arabidopsis. Furthermore, Thellungiella had higher amounts of several metabolites, such as soluble sugars and organic acids, under N-sufficient conditions (4 mm nitrate). Nitrate reductase activity and NR2 gene expression in Thellungiella displayed less of a reduction in response to N limitation than in Arabidopsis. Thellungiella shoot GS1 expression was more induced by low N than in Arabidopsis, while in roots, Thellungiella GS2 expression was maintained under N limitation but was decreased in Arabidopsis. Up-regulation of NRT2.1 and NRT3.1 expression was higher and repression of NRT1.1 was lower in Thellungiella roots under N-limiting conditions compared with Arabidopsis. Differential transporter gene expression was correlated with higher nitrate influx in Thellungiella at low (15)NO(3)(-) supply. Taken together, our results suggest that Thellungiella is tolerant to N-limited conditions and could act as a model system to unravel the mechanisms for low N tolerance.


Outcome Comparison between Open and Endovascular Management of TASC II D Aortoiliac Occlusive Disease.
Endovascular management of complex aortoiliac occlusive disease (AIOD) has been described as a viable alternative to open surgical reconstruction. To date, few studies have directly compared the 2 techniques. We therefore, evaluated short and mid- term outcomes of open and endovascular therapy in TASC II D AIOD patients. TASC II D patients undergoing treatment between January 2009 and December 2016 were retrospectively reviewed. Patient demographics, clinical data, and outcomes (complications [technical and systemic] and graft patency) were collected. The primary outcome of this study was primary graft patency. Patients were compared according to treatment group (open versus endovascular). Kaplan-Meier curves were used to analyze follow up results. A total of 75 consecutive patients (open: 30; endovascular: 45) were included in this analysis. In the endovascular group, 25 (55.6%) patients were managed using a hybrid approach with 100% technical success. Critical limb ischemia was the indication for intervention in 16.0% of this cohort (open, 13.3% vs. endovascular, 17.8%, P = 0.397). Overall, there were no significant differences in gender (male: open, 50.0% vs. endovascular, 55.6%, P = 0.637) or age (54.5 ± 5.9 years vs. 57.0 ± 8.7 years, P = 0.171). No in hospital deaths occurred in this cohort. The overall complication rate was significantly higher in the open group (43.3% vs. 17.8%, OR 3.5, 95% CI [1.2-10.1], P = 0.016) with peri-operative systemic complications being more likely in the open cohort (40.0% vs. 6.7%, OR 9.3, 95% CI [2.3-37.3], P < 0.001) while technical complications did not differ between the 2 groups (6.7% vs. 11.1%, OR 0.6, 95% CI [0.1-3.1], P = 0.517). Follow up data was available for 68 patients (90.7%), for a mean of 21.3 ± 17.1 months (range: 1-72 months). Re-intervention rates were significantly higher in the endovascular group (3.3% vs. 20.0%, OR 7.2, 95% CI [1.1-14.3], P = 0.038). The overall primary patency at 2 years was significantly higher in the open group (96.7% vs. 80.0%, OR 7.2, 95% CI [1.2-60.5], P = 0.038). Cox regression analysis revealed separation of the primary outcome for open therapy relative to endovascular repair (log rank, P = 0.320). In this comparison of open and endovascular therapy for complex AIOD, endovascular therapy was associated with high initial technical success and fewer in-hospital systemic complications but also high re-intervention rates when compared to open repair. Further prospective studies aimed at reduction of complications, optimization of patency, and patient selection for such procedures is warranted.


Putting the "mental" back in "mental disorders": a perspective from research on fear and anxiety.
Mental health problems often involve clusters of symptoms that include subjective (conscious) experiences as well as behavioral and/or physiological responses. Because the bodily responses are readily measured objectively, these have come to be emphasized when developing treatments and assessing their effectiveness. On the other hand, the subjective experience of the patient reported during a clinical interview is often viewed as a weak correlate of psychopathology. To the extent that subjective symptoms are related to the underlying problem, it is often assumed that they will be taken care of if the more objective behavioral and physiological symptoms are properly treated. Decades of research on anxiety disorders, however, show that behavioral and physiological symptoms do not correlate as strongly with subjective experiences as is typically assumed. Further, the treatments developed using more objective symptoms as a marker of psychopathology have mostly been disappointing in effectiveness. Given that "mental" disorders are named for, and defined by, their subjective mental qualities, it is perhaps not surprising, in retrospect, that treatments that have sidelined mental qualities have not been especially effective. These negative attitudes about subjective experience took root in psychiatry and allied fields decades ago when there were few avenues for scientifically studying subjective experience. Today, however, cognitive neuroscience research on consciousness is thriving, and offers a viable and novel scientific approach that could help achieve a deeper understanding of mental disorders and their treatment.


Does Embryo Culture Medium Influence the Health and Development of Children Born after In Vitro Fertilization?
In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No. 371) were randomized according to two media (Single Step Medium--SSM group) or Global medium (Global group). This randomized study was stopped prematurely as significantly lower pregnancy and implantation rates were observed in the SSM group. Singletons (No. 73) conceived in the randomized study were included (42 for Global and 31 for SSM). The medical data for gestational, neonatal and early childhood periods were extracted from medical records and parental interviews (256 variables recorded). The developmental profiles of the children in eight domains (social, self-help, gross motor, fine motor, expressive language, language comprehension, letter knowledge and number knowledge--270 items) were compared in relation to the culture medium. The delivery rate was significantly lower in the SSM group than in the Global group (p<0.05). The culture medium had no significant effect on birthweight, risk of malformation (minor and major), growth and the frequency of medical concerns. However, the children of the Global group were less likely than those of the SSM group to show developmental problems (p = 0.002), irrespective of the different domains. In conclusion, our findings showed that the embryo culture medium may have an impact on further development.


Comparison of ultrasound-guided fine-needle aspiration biopsy with core-needle biopsy in the evaluation of thyroid nodules.
To compare the diagnostic rate of ultra-sound-guided fine-needle aspiration biopsy (FNAB) with the diagnostic rate of combined FNAB and core-needle biopsy in the evaluation of nodular thyroid disease. We performed a retrospective case-control study by reviewing charts of patients who underwent ultra-sound-guided FNAB and core-needle biopsy of the thyroid at a tertiary referral center from January 1999 to December 2001. Results were classified as diagnostic (negative, suspicious, or positive for malignancy) or nondiagnostic. These findings were compared with an age- and sex-matched control group who underwent only FNAB. Complications between the groups were reviewed. The patient group consisted of 320 patients who underwent 340 ultrasound-guided fine-needle aspiration and core-needle biopsies of the thyroid; the control group consisted of 311 patients who underwent 340 FNABs. There was no significant difference in the nondiagnostic rates between groups--12.9% in patients who had FNAB-only compared with 10.9% in patients who had both procedures (proportion difference, -2.1%; 95% confidence interval, -7.0% to 2.9%; P = .41). There was a trend towards an increased incidence of hematoma and infection in the core biopsy group. In the group that underwent FNAB and core-needle biopsies, 10 patients (3.1%) developed biopsy-specific complications (hematomas in 8 patients, biopsy site infections in 2 patients). In the FNAB-only group, 3 patients (1.0%) developed hematomas; there was no incidence of infection. In the evaluation of thyroid nodules, the addition of core-needle biopsies to FNAB confers little benefit in decreasing the nondiagnostic rates and may be associated with increased complications. Core-needle biopsies should not be routinely performed in the evaluation of thyroid nodules, but rather, patient selection for the more invasive core biopsy should be done judiciously.


Effects of obesity and caloric intake on biliary lipid metabolism in man.
The effects of obesity and caloric intake on biliary lipid metabolism were investigated in a series of related studies. The degree of saturation of gallbladder bile with cholesterol was found to be significantly higher in a group of 23 obese healthy subjects than in a group of 23 nonobese controls matched for age, sex, and race. Bile was also significantly more saturated in 11 obese subjects before than after weight reduction. To determine whether supersaturated bile in obesity is due to excessive secretion of cholesterol or to deficient secretion of bile acids and phospholipids, the hepatic outputs of these three lipids were measured during constant duodenal infusion of formula in the same 11 subjects before and after weight reduction. Weight reduction resulted in significant reduction of cholesterol output but not of bile acid or phospholipid output. Moreover, very obese subjects were found to have cholesterol secretion rates markedly higher than less obese subjects previously studied by the same method. In obese subjects, bile was supersaturated with cholesterol despite increased bile acid pool sizes and increased secretion rates of bile acids and phospholipids. Supersaturated bile in the obese could therefore be attributed to a single defect in lipid secretion, namely, an excessive output of cholesterol. To determine whether the rate of caloric intake can account for the effects of obesity on biliary lipid composition and secretion, nine obese white men were studied on a weight maintenance diet and then during weight reduction on a 1,000 cal diet. As compared to weight maintenance, chronic caloric restriction resulted in reduced outputs of cholesterol, bile acids, and phospholipids, reduced bile acid pool size, and reduced synthesis and fecal excretion of cholesterol. Saturation of bile with cholesterol did not decrease during weight reduction, evidently because of the mobilization of cholesterol from adipose stores and the marked reduction in bile acid and phospholipid output observed during chronic caloric restriction. Acute alterations in caloric infusion rates did not fully reproduce the effects of chronic administration of high and low calorie diets. Likewise, chronic intake of hypercaloric diets by nonobese subjects did not reproduce the cholesterol hypersecretion characteristic of the obese. Thus, increased cholesterol secretion in obese subjects could not be fully explained by the amount of calories they ingested to maintain stable weight. It is concluded that obesity is characterized by excessive hepatic secretion of cholesterol which results in supersaturated bile.


Optically isotropic longitudinal piezoelectric resonant photoelastic modulator for wide angle polarization modulation at megahertz frequencies.
Polarization modulators have a broad range of applications in optics. The acceptance angle of a free-space polarization modulator is crucial for many applications. Polarization modulators that can achieve a wide acceptance angle are constructed by attaching a piezoelectric transducer to an isotropic material, and utilizing a resonant transverse interaction between light and acoustic waves. Since their demonstration in the 1960s, the design of these modulators has essentially remained the same with minor improvements in the following decades. In this work, we show that a suitable single crystal with the correct crystal orientation, functioning as both the piezoelectric transducer and the acousto-optic interaction medium, could be used for constructing a highly efficient free-space resonant polarization modulator operating at megahertz frequencies and exhibiting a wide acceptance angle. We construct the modulator using gallium arsenide, an optically isotropic and piezoelectric crystal, and demonstrate polarization modulation at 6 MHz with an input aperture of 1 cm in diameter, acceptance angle reaching ±30∘, and modulation efficiency exceeding 50%. Compared to state-of-the-art resonant photoelastic modulators, the modulator reported in this work exhibits greater than 50-fold improvement in modulation frequency for the same input aperture, while simultaneously reducing the thickness by approximately a factor of 80. Increasing the modulation frequency of photoelastic modulators from the kilohertz to the megahertz regime and substantially reducing their thickness lead to significant performance improvements for various use cases. This technological advancement also creates opportunities for utilizing these devices in new applications.


Aldosterone inhibition and cardiovascular protection: more important than it once appeared.
Aldosterone is present and active all along the cardiovascular continuum. Excessive tissue production occurs in cardiovascular diseases including myocardial infarction (MI) and heart failure, resulting in a multitude of adverse effects in the cardiovascular system necessitating pharmacologic blockade of this neurohormone. Both human and animal studies have consistently proven the beneficial effects of antialdosteronics in the improvement of: 1) endothelial function, 2) modulation of inflammatory mechanisms between blood and the vascular wall and 3) reduction of tissue proliferation and cardiovascular remodeling leading to different severities of cardiovascular damage. These basic mechanisms of anti-aldosterone therapy strongly support the promising data observed in major clinical trials with aldosterone blockers in cardiovascular diseases, specially in heart failure patients. Whereas aldosterone receptor blockers were initially viewed as potassium-sparing diuretics there has been a clear change of concept in the past 10 years, mainly following the positive results of RALES with spironolactone in chronic heart failure, followed by EPHESUS using eplerenone in patients with systolic dysfunction post MI. The significant positive results in both studies were a clear support for the inclusion of this pharmacologic intervention as first line treatment in most international guidelines for the management of heart failure. More recent and ongoing studies are exploring the usefulness of this type of intervention in preventing vascular and myocardial hypertrophy and remodeling in refractory hypertensive and some hyperfibrotic syndromes. There are also provocative studies investigating in the possibility of inhibiting atherosclerosis. More recently, some studies are suggesting the benefit of aldosterone blockade in sleep apnea. In addition, two large multicentric trials, TOPCAT and EMPHASIS are analyzing the potential use of antialdosteronics in patients with cardiac insufficiency and preserved systolic function and the possibility of extending their indication in systolic heart failure to Phase II respectively. New compounds, blocking the synthesis of aldosterone instead of blocking its receptor are being developed, and initial Phase 2 studies are positive. All of the above results are very interesting, show an optimistic future and are consolidating and enlarging the spectrum of aldosterone blockade in cardiovascular disorders every day.


Motor excitability is reduced prior to voluntary movements in children and adolescents with Tourette syndrome.
Tourette syndrome (TS) is a neuro-developmental disorder characterized by the occurrence of motor and vocal tics: involuntary, repetitive, stereotyped behaviours that occur with a limited duration, often typically many times in a single day. Previous studies suggest that children and adolescents with TS may undergo compensatory, neuroplastic changes in brain structure and function that help them gain control over their tics. In the current study we used single-pulse and dual-site paired-pulse transcranial magnetic stimulation (TMS), in conjunction with a manual choice reaction time task that induces high levels of inter-manual conflict, to investigate this conjecture in a group of children and adolescents with TS, but without co-morbid Attention Deficit Hyperactivity Disorder (ADHD). We found that performance on the behavioural response-conflict task did not differ between the adolescents with TS and a group of age-matched typically developing individuals. By contrast, our study demonstrated that cortical excitability, as measured by TMS-induced motor-evoked potentials (MEPs), was significantly reduced in the TS group in the period immediately preceding a finger movement. This effect is interpreted as consistent with previous suggestions that the cortical hyper-excitability that may give rise to tics in TS is actively suppressed by cognitive control mechanisms. Finally, we found no reliable evidence for altered patterns of functional inter-hemispheric connectivity in TS. These results provide evidence for compensatory brain reorganization that may underlie the increased self-regulation mechanisms that have been hypothesized to bring about the control of tics during adolescence.


Meaning in Life, Religious Coping, and Loneliness During the Coronavirus Health Crisis in Turkey.
Since December 2019, the COVID-19 pandemic has caused serious mental health challenges and consequently the Turkish population has been adversely affected by the virus. The present study examined how meaning in life related to loneliness and the degree to which religious coping strategies mediated these relations. Participants were a sample of 872 adults (242 males and 360 females) drawn from general public in Turkey. Data were collected using Meaning in Life Questionnaire, UCLA Loneliness Scale, and the Religious Coping Measure. Meaning in life was associated with more positive religious coping and less negative religious coping and loneliness. Positive religious coping was associated with less loneliness, while negative religious coping was associated with more loneliness. Religious coping strategies mediated the impact of meaning in life on loneliness. These findings suggest that greater meaning in life may link with lesser loneliness due to, in part, an increased level of positive religious coping strategies and a decreased level of negative coping strategies.


The Golgi Ca2+-ATPase KlPmr1p function is required for oxidative stress response by controlling the expression of the heat-shock element HSP60 in Kluyveromyces lactis.
The Golgi P-type Ca2+-ATPase, Pmr1p, is the major player for calcium homeostasis in yeast. The inactivation of KlPMR1 in Kluyveromyces lactis leads to high pleiotropic phenotypes that include reduced glycosylation, cell wall defects, and alterations of mitochondrial metabolism. In this article we found that cells lacking KlPmr1p have a morphologically altered mitochondrial network and that mitochondria (m) from Klpmr1delta cells accumulate Ca2+ more slowly and reach a lower [Ca2+]m level, when exposed to [Ca2+] < 5 microM, than wild-type cells. The Klpmr1delta cells also exhibit traits of ongoing oxidative stress and present hyperphosphorylation of KlHog1p, the hallmark for the activation of stress response pathways. The mitochondrial chaperone KlHsp60 acts as a multicopy suppressor of phenotypes that occur in cells lacking the Ca2+-ATPase, including relief from oxidative stress and recovery of cell wall thickness and functionality. Inhibition of KlPMR1 function decreases KlHSP60 expression at both mRNA and protein levels. Moreover, KlPRM1 loss of function correlates with both decreases in HSF DNA binding activity and KlHSP60 expression. We suggest a role for KlPMR1 in HSF DNA binding activity, which is required for proper KlHSP60 expression, a key step in oxidative stress response.


Clinicopathological features and immunohistochemical utility of NTRK-, ALK-, and ROS1-rearranged papillary thyroid carcinomas and anaplastic thyroid carcinomas.
NTRK1/3, ALK, and ROS1 translocations have been reported in a minor subset of papillary thyroid carcinomas (PTCs). We aimed to elucidate the prevalence and clinicopathological characteristics of these gene rearrangements and the utility of immunohistochemistry (IHC) in PTC and anaplastic thyroid carcinoma (ATC). We screened nonradiation-exposed cases of 307 PTCs and 16 ATCs by IHC for pan-Trk, ALK, and ROS1, followed by fluorescence in situ hybridization (FISH). In the PTC group, IHC for pan-Trk, ALK, and ROS1 was positive in 18 cases (5.9%), 1 case (0.3%), and 12 cases (3.9%), respectively. Among the pan-Trk IHC-positive cases (n = 18), 2 cases (11.1%; 0.7% of all PTCs) had NTRK1 or NTRK3 gene rearrangement with conventional PTC histology. The ALK IHC-positive case (n = 1) was the follicular variant of PTC with consistent ALK gene rearrangement. ROS1 gene rearrangement was not detectable in the ROS1 IHC-positive PTCs (0/12) by FISH. Most (approximately 70%) of the pan-Trk or ROS1 IHC-positive/FISH-negative cases had BRAF gene mutation with conventional PTC morphology. In the ATC group, neither ALK nor ROS1 IHC was positive, whereas pan-Trk IHC was positive in 1 case (6.3%) in which NTRK1 gene rearrangement was confirmed by FISH. These results suggest that NTRK, ALK, and ROS1 rearrangements are rare molecular events in nonradiation-exposed Japanese patients with PTC and ATC. Although IHC is not an entirely specific surrogate for these abnormalities and does not serve as a stand-alone companion diagnosis, the combined use of IHC and molecular testing may be helpful for determining promising therapeutic strategies with tyrosine kinase inhibitors.


The Application of Neoprene Diving Boots in Reduction of Feet Injuries in the Sowing Process Among the Rice Farmers in Thailand.
Foot injuries have been a problem among Thai farmers for a long time. The sowing process is one of the high-risk processes because farmers have to walk in the field to spray rice seed without footwear. Literature review showed that cuts from stepping on sharp material or snail shells were major sources of injury. Traditional footwear such as Ninja shoes or cloth socks have raised the questions regarding their ability in protection from such injuries. This study investigates using Neoprene diving boots to protect from foot injuries during the rice sowing process. The sample from this study were farmers from Sena District, Phra Nakhon Si Ayutthaya Province, Thailand. 29 farmer wearing Neoprene diving boots, 7 farmers wearing traditional Ninja shoes, and 19 farmers wearing cloth socks were asked to work in the sowing process and interviewed about any foot injuries that may have occurred. The statistical test showed there were no significant differences in characteristics among each group of farmers, except for work experience. The results show that there were no foot injuries in farmers wearing Neoprene diving boots, whereas 73% of farmers wearing Ninja shoes, and 14% of farmers wearing cloth socks still had foot injuries. Most injuries were alaceration that were caused by the golden apple snail. The satisfaction survey showed that farmers were satisfied and strongly satisfied with all of the aesthetics, utilization, and functionality factors. This study suggested that the Neoprene diving boots can be recommended for farmers to use as safeguarding against foot injuries in the rice sowing process.


Distribution of Health Resource Allocation in the Fars Province Using the Scalogram Analysis Technique in 2011.
The importance of health indicators in the recent years has created challenges in resource allocation. Balanced and fair distribution of health resources is one of the main principles in achieving equity. The goal of this cross-sectional descriptive study, conducted in 2010, was to classify health structural indicators in the Fars province using the scalogram technique. Health structural indicators were selected and classified in three categories; namely institutional, human resources, and rural health. The data were obtained from the statistical yearbook of Iran and was analyzed according to the scalogram technique. The distribution map of the Fars province was drawn using ArcGIS (geographic information system). The results showed an interesting health structural indicator map across the province. Our findings revealed that the city of Mohr with 85 and Zarindasht with 36 had the highest and the lowest scores, respectively. This information is valuable to provincial health policymakers to plan appropriately based on factual data and minimize chaos in allocating health resources. Based on such data and reflecting on the local needs, one could develop equity based resource allocation policies and prevent inequality. It is concluded that, as top priority, the provincial policymakers should place dedicated deprivation programs for Farashband, Eghlid and Zaindasht regions.


Highly Variable Disease Courses in Siblings with Stargardt Disease.
To investigate intersibling phenotypic concordance in Stargardt disease (STGD1). Retrospective cohort study. Siblings with genetically confirmed STGD1 and at least 1 available fundus autofluorescence (FAF) image of both eyes. We compared age at onset within families. Disease duration was matched to investigate differences in best-corrected visual acuity (BCVA) and compared the survival time for reaching severe visual impairment (<20/200 Snellen or >1.0 logarithm of the minimum angle of resolution [logMAR]). Central retinal atrophy area was quantified independently by 2 experienced graders using semiautomated software and compared between siblings. Both graders performed qualitative assessment of FAF and spectral-domain (SD) OCT images to identify phenotypic differences. Differences in age at onset, disease duration-matched BCVA, time to severe visual impairment development, FAF atrophy area, FAF patterns, and genotypes. Substantial differences in age at onset were present in 5 of 17 families, ranging from 13 to 39 years. Median BCVA at baseline was 0.60 logMAR (range, -0.20 to 2.30 logMAR; Snellen equivalent, 20/80 [range, 20/12-hand movements]) in the right eye and 0.50 logMAR (range, -0.20 to 2.30 logMAR; Snellen equivalent, 20/63 [range, 20/12-hand movements]) in the left eye. Disease duration-matched BCVA was investigated in 12 of 17 families, and the median difference was 0.41 logMAR (range, 0.00-1.10 logMAR) for the right eye and 0.41 logMAR (range, 0.00-1.08 logMAR) for the left eye. We observed notable differences in time to severe visual impairment development in 7 families, ranging from 1 to 29 years. Median central retinal atrophy area was 11.38 mm2 in the right eye (range, 1.98-44.78 mm2) and 10.59 mm2 in the left eye (range, 1.61-40.59 mm2) and highly comparable between siblings. Similarly, qualitative FAF and SD OCT phenotypes were highly comparable between siblings. Phenotypic discordance between siblings with STGD1 carrying the same ABCA4 variants is a prevalent phenomenon. Although the FAF phenotypes are highly comparable between siblings, functional outcomes differ substantially. This complicates both sibling-based prognosis and genotype-phenotype correlations and has important implications for patient care and management.


Perception of facial affect in chronic schizophrenia and right brain damage.
This study was designed to compare the performance of 50 chronic schizophrenics (CS) to that of 30 right brain-damaged patients (R), and 50 healthy controls (N) on several facial perception measures: Emotion Labelling and Recognition, and the Benton Facial Recognition Test. CSs were diagnosed according to DSM-IV criteria and their psychiatric state was assessed using the PANSS scale. All subjects were right handed. Their cognitive state was assessed using the MMSE. Subjects rated their current mood on a visual analogue scale. The results showed that the CSs and Rs were significantly impaired compared to Ns for the emotional tasks but did not differ from each other. Moreover, the patient groups were significantly less accurate in recognising emotionally neutral facial stimuli. Each subject group had more difficulty processing negative relative to positive affect. The deficit in schizophrenia was found to be stable, which may reflect a trait-like, rather than a state-dependent, characteristic. Moreover, some support is provided for the notion that facial affect perception in chronic schizophrenia is associated with right hemisphere dysfunction.


Coping behavior in depressed patients: a longitudinal study.
The relationship of coping behavior to outcome in depressed patients was examined. Subjects (n=105) with major depressive disorder (n=85), depressive disorder not otherwise specified (n=7) or major depressive disorder with axis I comorbidity (n=13) were followed for 6 months. Their coping behavior (i.e. rumination, active distraction, cognitive distraction and dangerous activities) was defined using the Comprehensive Assessment List for Affective Disorders. Based on their Hamilton Rating Scale for Depression (HRSD) scores at 6 months, the patients were categorized as having had a good or a poor outcome. Severity of depression and coping behavior were similar among the three diagnostic groups. At baseline assessment, coping behavior was not correlated with either HRSD score or age. However, males were significantly more likely to be engaged in dangerous activity as a coping behavior than females. Patients with a good outcome at 6 months were significantly more likely to use rumination as a coping behavior while patients with a poor outcome were significantly more likely to use dangerous activity. Multiple regression analysis confirmed this finding, indicating that rumination and dangerous activity were significant predictors of outcome at 6 months. Rumination might be associated with good outcomes in depressed patients while dangerous activity might be associated with poor outcomes.


Understanding nitrate reactions with zerovalent iron using tafel analysis and electrochemical impedance spectroscopy.
This study investigated the reaction mechanisms of nitrate (NO3-) with zerovalent iron (ZVI) media under conditions relevantto groundwatertreatment using permeable reactive barriers (PRB). Reaction rates of NO3- with freely corroding and with cathodically or anodically polarized iron wires were measured in batch reactors. Tafel analysis and electrochemical impedance spectroscopy (EIS) were used to investigate the reactions occurring on the iron surfaces. Reduction of NO3- by corroding iron resulted in near stoichiometric production of NO2-, which did not measurably react in the absence of added Fe(II). Increasing NO3- concentrations resulted in increasing corrosion currents. However, EIS and Tafel analyses indicated that there was little direct reduction of NO3- at the ZVI surface, despite the presence of water reduction. This behavior can be attributed to formation of a microporous oxide on the iron surfaces that blocked reduction of NO3- and NO2- but did not block water reduction. This finding is consistent with previous observations that NO3- impedes reduction of organic compounds by ZVI. Nitrite concentrations greater than 4 mM resulted in anodic passivation of the iron, but passivation was not observed with NO3- concentrations as high as 96 mM. This indicates that the passivating oxide preventing NO3- reduction was permeable toward cation migration. Since reaction with Fe(0) can be excluded asthe mechanism for NO3- and NO2- reduction, reaction with Fe(II)-containing oxides coating the iron surface is the most likely reaction mechanism. This suggests that short-term batch tests requiring little turnover of reactive sites on the iron surface may overestimate long-term rates of NO3- removal because the effects of passivation are not apparent in batch tests conducted with high initial Fe(II) to NO3- ratios.


Comparison of the salivary and dentinal microbiome of children with severe-early childhood caries to the salivary microbiome of caries-free children.
The main objectives of this study were to describe and compare the microbiota of 1) deep dentinal lesions of deciduous teeth of children affected with severe early childhood caries (S-ECC) and 2) the unstimulated saliva of these children and 3) the unstimulated saliva of caries-free children, and to compare microbiota compositional differences and diversity of taxa in these sampled sites. Children with S-ECC and without S-ECC were recruited. The saliva of all children with and without S-ECC was sampled along with the deep dentinal microbiota from children affected by S-ECC. The salivary microbiota of children affected by S-ECC (n = 68) was compared to that of caries-free children (n = 70), by Illumina MiSeq sequencing of 16S rRNA amplicons. Finally, the caries microbiota of deep dentinal lesions of those children with S-ECC was investigated. Using two beta diversity metrics (Bray Curtis dissimilarity and UniFrac distance), the caries microbiota was found to be distinct from that of either of the saliva groups (caries-free & caries-active) when bacterial abundance was taken into account. However, when the comparison was made by measuring only presence and absence of bacterial taxa, all three microbiota types separated. While the alpha diversity of the caries microbiota was lowest, the diversity difference between the caries samples and saliva samples was statistically significant (p < 0.001). The major phyla of the caries active dentinal microbiota were Firmicutes (median abundance value 33.5%) and Bacteroidetes (23.2%), with Neisseria (10.3%) being the most abundant genus, followed by Prevotella (10%). The caries-active salivary microbiota was dominated by Proteobacteria (median abundance value 38.2%) and Bacteroidetes (27.8%) with the most abundant genus being Neisseria (16.3%), followed by Porphyromonas (9.5%). Caries microbiota samples were characterized by high relative abundance of Streptococcus mutans, Prevotella spp., Bifidobacterium and Scardovia spp. Distinct differences between the caries microbiota and saliva microbiota were identified, with separation of both salivary groups (caries-active and caries-free) whereby rare taxa were highlighted. While the caries microbiota was less diverse than the salivary microbiota, the presence of these rare taxa could be the difference between health and disease in these children.


[Interactions between Neisseria meningitidis and cell cultures (data of cytomorphologic studies)].
The present work shows that the cytopathogenic action of N. meningitidis on continuous human amniotic epithelial cell culture FL begins from their active adhesion and subsequent invasion. The degree of the manifestation of the cytopathic effect depends on the capacity of the infective agent for adhesion and invasion and on its biological properties, as well as on the initial state of the cells. The infection of the cells is accompanied by disturbances in their mitotic activity together with the lesions of their chromosomal apparatus. The cells die either in the state of degenerative mitosis, or as the result of the rupture of the cytoplasm in massively invaded cells. The response of the cells to the invasion of faintly cytopathogenic and noncytopathogenic strains takes the form of nonprofessional phagocytosis.


Noninvasive study of high-energy phosphate metabolism in human heart by depth-resolved 31P NMR spectroscopy.
Phosphorus-31 nuclear magnetic resonance (NMR) spectra showing the relative concentrations of high-energy phosphate metabolites have been recorded noninvasively from the human heart in vivo. Spectral data were spatially localized by combining a pulsed magnetic field gradient with surface NMR excitation-detection coils. The location of the selected spectral region was determined by conventional proton NMR imaging immediately before examination by phosphorus-31 NMR spectroscopy.


Effect of Atmospheric CO2 Enrichment on Root Growth and Carbohydrate Allocation of Phaseolus spp.
A glasshouse experiment was conducted with plants of Phaseolus grown in liquid culture. Root growth parameters (biomass, diameter, length, growth rate, zone of cell division), root rheological components (wall extensibility, water potential yield threshold, water potential), shoot growth, carbon allocation, and abscisic acid (ABA) concentration were measured in Phaseolus acutifolius A. Gray at ambient (550 µmol mol-1) and elevated (700 µmol mol-1) atmospheric CO2 concentrations. For contrast, measurements of above- and belowground growth were conducted on Phaseolus vulgaris L. in the same treatments. Under nonlimiting conditions of water and nutrients, elevated CO2 increased root and shoot growth of P. acutifolius but not P. vulgaris. While root mass was increased by nearly 60% in P. acutifolius, there was no effect of atmospheric CO2 on any of the rheological components measured. In contrast, starch and ABA accumulated in roots of P. acutifolius. The concentration of starch in roots of P. acutifolius increased by 10-fold, while root concentrations of ABA doubled. From the data it is concluded that CO2 enrichment is favorable for root growth in some species in that more carbon is allocated to belowground growth. In addition, ABA may play a role in growth responses and/or allocation of photosynthates at elevated CO2 in P. acutifolius.


Heterotrophic Proteobacteria in the vicinity of diffuse hydrothermal venting.
Deep-sea hydrothermal vents are highly dynamic habitats characterized by steep temperature and chemical gradients. The oxidation of reduced compounds dissolved in the venting fluids fuels primary production providing the basis for extensive life. Until recently studies of microbial vent communities have focused primarily on chemolithoautotrophic organisms. In our study, we targeted the change of microbial community compositions along mixing gradients, focusing on distribution and capabilities of heterotrophic microorganisms. Samples were retrieved from different venting areas within the Menez Gwen hydrothermal field, taken along mixing gradients, including diffuse fluid discharge points, their immediate surroundings and the buoyant parts of hydrothermal plumes. High throughput 16S rRNA gene amplicon sequencing, fluorescence in situ hybridization, and targeted metagenome analysis were combined with geochemical analyses. Close to diffuse venting orifices dominated by chemolithoautotrophic Epsilonproteobacteria, in areas where environmental conditions still supported chemolithoautotrophic processes, we detected microbial communities enriched for versatile heterotrophic Alpha- and Gammaproteobacteria. The potential for alkane degradation could be shown for several genera and yet uncultured clades. We propose that hotspots of chemolithoautotrophic life support a 'belt' of heterotrophic bacteria significantly different from the dominating oligotrophic microbiota of the deep sea.


[Factor analysis of the results of combined clinico-radiological examination in thyroid diseases].
The paper is concerned with factor analysis data on the results of combined radioclinical investigation in thyroid diseases. A program including 111 parameters in 291 patients (anamnestic data, results of physical examination, palpation of the thyroid and the neck region, scannographic, thermographic, and ultrasound symptoms, indicators of radiometry and in vitro tests) was developed at the IBM Company. This method was employed for obtaining objective information on the diagnostic value of each method as well as for detecting certain symptoms of the greatest diagnostic importance in different nosological entities of thyroid diseases.


Both Ox1R and Ox2R orexin receptors contribute to the cardiorespiratory response evoked from the perifornical hypothalamus.
Orexin/hypocretin neurons are located in and around the perifornical hypothalamus. Disinhibition of this area in the anaesthetized preparation evokes cardiorespiratory changes that can be reduced to nearly half or more by systemic Almorexant, a dual receptor antagonist of the two known orexin receptors, Ox1R and Ox2R. It is not clear if these reductions result from the blockade of one receptor or both. To determine the contribution of the two receptors, we compared the effects of Almorexant to those of the selective Ox1R antagonist ACT335827 and the selective Ox2R antagonists EMPA and TCS-OX2-29. Bicuculline (20 pmol) was injected in the perifornical hypothalamus of urethane-anaesthetized rats before and after administration of the drugs (all 15 mg/kg, intravenously). The pressor, tachycardic and tachypneic responses to bicuculline were attenuated/reduced by ACT335827 (by 19%, ns; 10%, ns and 24%, P < 0.01, respectively), EMPA (by 35% P < 0.01; 6%, ns; and 26% P < 0.05) and TCS-OX2-29 (by 13%, ns; 10%, ns and 42%, P < 0.001). These reductions represented only a fraction of the reduction after Almorexant (by 43%, P < 0.001; 42%, P < 0.001 and 65% P < 0.001). However, when the selective Ox1R and Ox2R antagonists were given in combination, the reductions were greater and closer to those of Almorexant (ACT335827 + EMPA, by 26%, P < 0.05; 24%, P < 0.05 and 47%, P < 0.001; ACT335827 + TCS-OX2-29, by 40%, P < 0.01; 26%, P < 0.001 and 59%, P < 0.0001). This was particularly clear with the tachypneic response. These results suggest that both orexin receptors contribute to the cardiorespiratory response evoked from the hypothalamus under anaesthesia. They are consistent with our previous study in the conscious animal.


Human antibodies specific for the high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae mediate opsonophagocytic activity.
The HMW1- and HMW2-like adhesion proteins of nontypeable Haemophilus influenzae are expressed by 75% of these strains, and antibodies directed against these proteins are protective in animal models of infection. The purpose of the present study was to define the functional activity of human antibodies specific for these proteins in an in vitro complement-dependent opsonophagocytic assay. Human promyelocytic cell line HL-60 served as the source of phagocytic cells, and a commercial preparation of intravenous immunoglobulin (IVIG) served as the source of human antibodies. High-molecular-weight (HMW) proteins were purified from four prototype nontypeable H. influenzae strains and used to prepare solid-phase affinity columns. IVIG was adsorbed on each column to remove strain-specific anti-HMW antibodies and to allow recovery of affinity-purified anti-HMW antibody fractions. Unadsorbed IVIG killed each of the prototype strains at titers of 1:80 to 1:320. HMW-adsorbed sera demonstrated fourfold decreases in opsonophagocytic titer against the homologous strains compared to unadsorbed IVIG. Affinity-purified anti-HMW antibody preparations demonstrated opsonophagocytic titers of 1:20 to 1:80 against the respective homologous strains and opsonophagocytic titers as high as 1:80 against heterologous strains. None of the affinity-purified anti-HMW antibody preparations was opsonophagocytic for a representative nontypeable H. influenzae strain that did not express HMW1- or HMW2-like proteins. These data demonstrate that human antibodies specific for the HMW1/HMW2-like adhesion proteins of nontypeable H. influenzae are opsonophagocytic and that such antibodies recognize epitopes shared by the HMW proteins of unrelated nontypeable H. influenzae strains. These results argue for continued investigation of the HMW1/HMW2-like proteins as potential vaccine candidates for prevention of disease due to nontypeable H. influenzae.


Testing Nonmonotonicity in Health Preferences.
The main aim of this article is to test monotonicity in life duration. Previous findings suggest that, for poor health states, longer durations are preferred to shorter durations up to some threshold or maximum endurable time (MET), and shorter durations are preferred to longer ones after that threshold. Monotonicity in duration is tested through 2 ordinal tasks: choices and rankings. A convenience sample (n = 90) was recruited in a series of experimental sessions in which participants had to rank-order health episodes and to choose between them, presented in pairs. Health episodes result from the combination of 7 EQ-5D-3L health states and 5 durations. Monotonicity is tested comparing the percentage rate of participants whose preferences were monotonic with the percentage of participants with nonmonotonic preferences for each health state. In addition, to test the existence of preference reversals, we analyze the fraction of people who switch their preference from rankings to choices. Monotonicity is frequently violated across the 7 EQ-5D health states. Preference patterns for individuals describe violations ranging from almost 49% with choices to about 71% with rankings. Analysis performed by separate states shows that the mean rates of violations with choices and ranking are about 22% and 34%, respectively. We also find new evidence of preference reversals and some evidence-though scarce-of transitivity violations in choices. Our results show that there is a medium range of health states for which preferences are nonmonotonic. These findings support previous evidence on MET preferences and introduce a new "choice-ranking" preference reversal. It seems that the use of 2 tasks with a similar response scale may make preference reversals less substantial, although it remains important and systematic. Two procedures based on ordinal comparisons are used to elicit preferences: direct choices and rankings. Our study reports significant rates of nonmonotonic preferences (or maximum endurable time [MET]-type preferences) for different combinations of durations and EQ-5D health states.Analysis for separate health states shows that the mean rates of nonmonotonicity range from 22% (choices) to 34% (rankings), but within-subject analysis shows that nonmonotonicity is even higher, ranging from 49% (choices) to 71% (rankings). These violations challenge the validity of multiplicative QALY models.We find that the MET phenomenon may affect particularly those EQ-5D health states that are in the middle of the severity scale and not so much the extreme health states (i.e., very mild and very severe states).We find new evidence of preference reversals even using 2 procedures of a similar (ordinal) nature. Percentage rates of preference reversals range from 1.5% to 33%. We also find some (although scarce) evidence on violations of transitivity.


Stab wounds of the brain.
Unlike the penetrating injuries to the brain caused by missiles, injuries by stabbing are largely restricted to the wound tract. Early recognition, debridement and judicious antibiotic therapy can limit or prevent complications in the management of stab wounds. Among the common sequelae of stab wounds of the brain are pneumocephalus, meningitis, intracerebral hemorrhage and direct blood vessel or nerve injury.


Nutritional programming of gastrointestinal tract development. Is the pig a good model for man?
The consequences of early-life nutritional programming in man and other mammalian species have been studied chiefly at the metabolic level. Very few studies, if any, have been performed in the gastrointestinal tract (GIT) as the target organ, but extensive GIT studies are needed since the GIT plays a key role in nutrient supply and has an impact on functions of the entire organism. The possible deleterious effects of nutritional programming at the metabolic level were discovered following epidemiological studies in human subjects, and confirmed in animal models. Investigating the impact of programming on GIT structure and function would need appropriate animal models due to ethical restrictions in the use of human subjects. The aim of the present review is to discuss the use of pigs as an animal model as a compromise between ethically acceptable animal studies and the requirement of data which can be interpolated to the human situation. In nutritional programming studies, rodents are the most frequently used model for man, but GIT development and digestive function in rodents are considerably different from those in man. In that aspect, the pig GIT is much closer to the human than that of rodents. The swine species is closely comparable with man in many nutritional and digestive aspects, and thus provides ample opportunity to be used in investigations on the consequences of nutritional programming for the GIT. In particular, the 'sow-piglets' dyad could be a useful tool to simulate the 'human mother-infant' dyad in studies which examine short-, middle- and long-term effects and is suggested as the reference model.


Distribution of killer cell immunoglobulin-like receptors (KIR) and their HLA-C ligands in two Iranian populations.
Killer cell immunoglobulin-like receptors (KIR) gene frequencies vary between populations and contribute to functional variation in immune responses to viruses,autoimmunity and reproductive success. This study describes the frequency distribution of 12 variable KIR genes and their HLA-C ligands in two Iranian populations who have lived for many generations in different environments:t he Azerbaijanis at high altitude and the Jonobi people at sea level. The results are compared with those published for other human populations and a large group of English Caucasians. Differences were seen in KIR and HLA-C group frequencies, in linkage disequilibrium and inhibitory/activating KIR ratios between the groups. Similarities with geographically close populations in the frequencies of the KIR A and B haplotypes and KIR AA genotype reflected their common ancestry. The extreme variability of the KIR gene family and their HLA-C ligands is highlighted and their importance in defining differences between geographically and culturally isolated communities subject to different environmental pressures who come from the same ethnic grouping.


Improving diabetes outcomes using a web-based registry and interactive education: a multisite collaborative approach.
To support the adoption of guideline concordant care by primary care practices, the New York Diabetes Coalition (NYDC) promoted use of an electronic diabetes registry and developed an interactive educational module on using the registry and improving patient communication. The NYDC hypothesized that use of a registry with immediate feedback would achieve measurable and clinically meaningful improvement in the proportion of patients at goal for diabetes health metrics. In 2006-2007, the NYDC recruited 7 small to midsized primary care practices to implement the registry and to receive education and coaching on registry use, practice work flow, and patient engagement. The patient cohort included those with 2 or more visits with a diagnosis of diabetes within a 12-month period. Each patient's health measure status (at goal, above goal, not recorded) was assessed quarterly for hemoglobin A1C , low-density lipoprotein (LDL), and blood pressure (BP), and most recent A1C value was noted. A cohort analysis was performed using random effects regression models to assess the impact of the registry over time for each diabetes health metric. After controlling for variability between sites, with each subsequent quarter during the registry period patients were 1.4 times more likely to have A1C ≤ 9, almost twice (OR = 1.8) as likely to have LDL < 100, and 1.3 times more likely to have BP < 140/90. These improvements in compliance were statistically significant. Average A1C also improved over time, though this did not reach statistical significance. Utilizing a Web-based registry and interactive education, the project demonstrated improved patient outcomes, as well as the feasibility of collecting aggregate data from unrelated, independent practices.


Value of information methods for assessing a new diagnostic test.
Value-of-information methods are applied to assess the evidence in support of a new diagnostic test and, where the evidence is insufficient for decision making, to determine the optimal sample size for future studies. Net benefit formulations are derived under various diagnostic and treatment scenarios. The expressions for the expected opportunity loss of adopting strategies that include the new test are given. Expressions for the expected value of information from future studies are derived. One-sample and two-sample designs, with or without known prevalence, are considered. An example is given.


Pseudomeningocele in communication with the facet joint: demonstration by computerized tomography-arthrography.
Pseudomeningocele is an uncommon but well-known complication of lumbar spine operations. Although it is mostly asymptomatic and managed conservatively in most cases, it is claimed as a causative factor of failed back surgery syndrome and requires surgery in some cases. Usually, its diagnosis is confidently done with imaging modalities such as magnetic resonance imaging, computed tomography and myelography. In this report, we describe a case of pseudomeningocele that communicated with a facet joint. The diagnostic approach for this unusual lesion and its probable causes are discussed.


Object dependency of resolution in reconstruction algorithms with interiteration filtering applied to PET data.
In this paper, we study the resolution properties of those algorithms where a filtering step is applied after every iteration. As concrete examples we take filtered preconditioned gradient descent algorithms for the Poisson log likelihood for PET emission data. For nonlinear estimators, resolution can be characterized in terms of the linearized local impulse response (LLIR). We provide analytic approximations for the LLIR for the class of algorithms mentioned above. Our expressions clearly show that when interiteration filtering (with linear filters) is used, the resolution properties are, in most cases, spatially varying, object dependent and asymmetric. These nonuniformities are solely due to the interaction between the filtering step and the Poisson noise model. This situation is similar to penalized likelihood reconstructions as studied previously in the literature. In contrast, nonregularized and postfiltered maximum-likelihood expectation maximization (MLEM) produce images with nearly "perfect" uniform resolution when convergence is reached. We use the analytic expressions for the LLIR to propose three different approaches to obtain nearly object independent and uniform resolution. Two of them are based on calculating filter coefficients on a pixel basis, whereas the third one chooses an appropriate preconditioner. These three approaches are tested on simulated data for the filtered MLEM algorithm or the filtered separable paraboloidal surrogates algorithm. The evaluation confirms that images obtained using our proposed regularization methods have nearly object independent and uniform resolution.


Serum Cortisol Level in Indian Patients with Severe Sepsis/Septic Shock.
The relationship between cortisol level and sepsis is not known in Indian patients of severe sepsis/septic shock. The study was done to determine the optimal range of cortisol levels, defining the adrenocortical response, and predicting the mortality, if possible, in the above type of patients. The study was a single-centered prospective cohort study, conducted in a tertiary referral center, North India. Sixty patients with severe sepsis (n = 30) and septic shock (n = 30) were recruited. Basal and postcosyntropin (1 μg)-stimulated cortisol levels were measured, and all patients were closely monitored with daily assessments of clinical and laboratory variables. Western diagnostic criteria were followed for defining adrenal insufficiency (AI). The end point was the survival assessed at day 28 or death, whichever came earlier. The mean basal (T0) and poststimulation (T30) cortisol levels were 31.77 ± 15.9 μg/dL and 37.58 ± 17.31 μg/dL, respectively. In all sepsis patients, 48.33% qualified as AI at T0 ≤ 24 μg/dL, 61.67% at delta cortisol (Δ = T30-T0) ≤7 μg/dL, and 78.33% at Δ ≤9 μg/dL. Using receiver operating characteristic curve, the area under the curve (AUC) was 0.4954, signifying poor prediction to death. Indians have completely different characteristics of cortisol levels in sepsis patients, in comparison to the Western data. They have higher range of basal cortisol levels, higher percentage of AI, and an inability to predict mortality with the cortisol levels. Hence, there is requirement of an international study to confirm the dichotomy of the results.


Simulation-based comparison of Biopharmaceutics Classification System and drug structure.
Background: The Biopharmaceutics Classification System (BCS), which classifies bioactive molecules based on solubility and permeability, is widely used to guide new drug development and drug formulation, as well as predict pharmacokinetics. Here we performed computer simulations to study correlations between a molecule's structure and its BCS classification. Methods: A total of 411 small molecules were assigned to BCS categories based on published drug data, and their Pybel-FP4 fingerprints were extrapolated. The information gain (IG) of each fingerprint was calculated and its characteristic structure analyzed. IG was calculated using multiple thresholds, and results were verified using support vector machine prediction, while taking into account the dose coefficient (0-0.1, 0.1-1, or >1). Structural functional features common to fingerprints of compounds in each type of BCS class were determined using computer simulations. Results: BCS classes III and IV appear to share several structural and functional characteristics, including secondary aliphaticamine, Michael acceptor, isothiourea, and sulfonamide sulfonic derivatives. Conclusion: We demonstrate that our approach can correlate characteristic fingerprints of small-molecule drugs with BCS classifications, which may help guide the development and optimization of new drugs.


[Diagnosis of uterine vascular malformation using Doppler ultrasound].
We present the case of a 56-year-old woman, who was admitted to our clinic for diagnostic laparoscopy because of a cystic uterine tumour of uncertain dignity. In the patient's history three curettages due to recurrent acyclic premenopausal vaginal bleeding were reported without specific histological findings. The preceding MRI described the structure as a myoma. During preoperative diagnostics an arteriovenous malformation was suspected by transvaginal Doppler sonography. Consequently the procedure was changed and a laparotomy performed. The sonographic findings were confirmed during surgery and by histological examination. This case points out the important role of transvaginal sonography combined with colour-flow-mapping. By confirming the diagnosis preoperatively and changing the management a low-risk procedure could be ensured.


Mandibular movement patterns during speech in subjects with temporomandibular disorders and in asymptomatic individuals.
The mandibular movements used during speech modify space to allow different articulation postures proper for each sound. Temporomandibular disorders (TMD) may cause modifications in these movements due to joint and muscular conditions. The aim of this study was to verify the amplitude and the characterization of the mandibular movements during speech, using computerized electrognathography, in individuals with TMD and in asymptomatic individuals, analyzing possible interferences of these dysfunctions. One hundred thirty-five (135) adult subjects were divided into two groups: GI with 90 participants diagnosed with TMD and GIII with 45 asymptomatic participants. Their mandibular movements were observed during the sequential naming of pictures containing all of the word sounds, which occur in the Brazilian Portuguese language. The records were obtained with computerized electrognathography (BioEGN-BioPak system, BioResearch Associates, Inc., Milwaukee, WI). Mean values of the amplitude were described for the two groups. The analysis of such results showed statistically significant differences between the means of the values, obtained for the two groups in the opening and retrusion ranges. Statistically significant differences were not established for the presence and the range of the deviations in laterality, during speech. Prevalence of bilateral deviations was verified in GIII and unilateral deviations in GI. This study describes the 3-dimensional thresholds of mandibular movements in speech for Brazilian Portuguese, for the investigated individuals of both groups. The presence of TMD shows reduction in mandibular opening and retrusion ranges and prevalence of unilateral deviation movements during speech.


A quantitative model of thermal stabilization and destabilization of proteins by ligands.
Equilibrium binding ligands usually increase protein thermal stability by an amount proportional to the concentration and affinity of the ligand. High-throughput screening for the discovery of drug-like compounds uses an assay based on thermal stabilization. The mathematical description of this stabilization is well developed, and the method is widely applicable to the characterization of ligand-protein binding equilibrium. However, numerous cases have been experimentally observed where equilibrium binding ligands destabilize proteins, i.e., diminish protein melting temperature by an amount proportional to the concentration and affinity of the ligand. Here, we present a thermodynamic model that describes ligand binding to the native and unfolded (denatured) protein states explaining the combined stabilization and destabilization effects. The model also explains nonsaturation and saturation effects on the protein melting temperature when the ligand concentration significantly exceeds the protein concentration. Several examples of the applicability of the model are presented, including specific sulfonamide binding to recombinant hCAII, peptide and ANS binding to the Polo-box domain of Plk1, and zinc ion binding to the recombinant porcine growth hormone. The same ligands may stabilize and destabilize different proteins, and the same proteins may be stabilized and destabilized by different ligands.


Three-dimensional echocardiography reveals early mitral valve alterations in hypertrophic cardiomyopathy genetic mutation carriers.
The mitral valve undergoes structural modifications in response to cardiac functional changes, often predating cardiac decompensation and overt clinical signs. Our study assessed the potential of mitral valve morphological changes as early indicators for detecting carriers of hypertrophic cardiomyopathy (HCM)-associated gene mutations. We studied 505 participants: 189 without the pathogenic gene mutations and left ventricular hypertrophy (G-/LVH-), 149 carriers without LV hypertrophy (G+/LVH-), and 167 manifest HCM patients (G+/LVH+). We juxtaposed the mitral valve morphology and associated metrics across these groups, emphasizing those carrying MYH7 and MYBPC3 mutations. We discerned pronounced disparities in the mitral annulus and leaflet structures across the groups. The mitral valve apparatus in mutation carriers exhibited a tendency towards a flattened profile. Detailed analysis spotlighted MYBPC3 mutation carriers, whose mitral valves were notably flatter (with notably lower AHCWR values than non-carriers); this contrast was not evident in MYH7 mutation carriers. This mitral valve flattening, manifest in the mutation carriers, suggests it might be an adaptive response to incipient cardiac dysfunction in HCM's nascent stages. Three-dimensional echocardiography illuminates the initial mitral valve structural changes in HCM patients bearing pathogenic gene mutations. These morphological signatures hold promise as sensitive imaging markers, especially for asymptomatic carriers of the MYBPC3 mutation.


Can cranberry extract and vitamin C + Zn supplements affect the in vivo activity of paraoxonase 1, antioxidant potential, and lipid status?
The modern way of life exposes us to substantial oxidative stress, putting the focus on the research of antioxidant effects of dietary supplements. Recent studies have shown that the effectiveness of particular vitamins and herbal preparations might have an effect on paraoxonase activity. Paraoxonase 1 is an HDL associated enzyme which prevents the oxidation of LDL. Several studies have shown the beneficial effect of some dietary components to the activity of paraoxonase. The aim of this study was to analyze the effects of cranberry extract and vitamin C and zinc preparations (vitamin C + Zn) on serum paraoxonase 1 activity, antioxidant status, and glucose and lipid concentration. The study included 31 healthy volunteers (median age 24 years). They were divided into 3 groups according to the intervention type and smoking status and exposed to commercially available preparations of the cranberry extract (2 g/day) and vitamin C + Zn (300 mg/day) during 4 weeks. The results have shown that there is a significant increase in the activity of the paraoxonase 1 in nonsmokers after the intervention with the cranberry extract as well as with vitamin C + Zn preparations. Also, total antioxidant status increased in the non-smokers subgroup after intervention with vitamin C + Zn. However, the lipid profile did not change significantly in response to antioxidant preparations. Our results show that antioxidant supplements can increase the antioxidant potential of an organism as well as paraoxonase 1 activity. This observation is pointing to the potential complementary role of dietary supplements in the primary prevention of atherosclerosis.


Lumbosacral transitional vertebra in 14 dog breeds in Norway: Occurrence, risk factors and association with hip dysplasia.
A lumbosacral transitional vertebra (LTV) is a congenital anomaly of the spine and has been suggested to predispose to canine hip dysplasia (CHD). This retrospective, cross-sectional study investigated the prevalence of LTV and CHD among 14 dog breeds in Norway, the possible associations with risk factors, and whether LTV was a risk factor for the development of hip dysplasia. The results were based on evaluation of ventrodorsal radiographs from the CHD screening program from the Norwegian Kennel Club from February 2014 to January 2022. A total of 13,950 dogs were included in the study. For statistical analysis, CHD grades were reclassified from the official Federation Cynologique Internationale (FCI) grades into three grades: CHD free (CHD=A, B), CHD mild (CHD=C), and severe (CHD=D, E). In the study sample, the overall occurrence of LTV was 18.5%, of which 32.9% were type 1, 45.7% type 2 and 21.4% type 3. The occurrence of LTV varied significantly among the included breeds, ranging from 9.5% to 46.2%. There was no association between sex and LTV. The frequencies of CHD grades were A: 43.1%; B: 31.4%; C: 18.4%; D: 6.0%; E: 1.1%. There was a statistically significant association with mild and severe CHD in dogs with LTV type 2 and LTV type 3 (P<0.001). In the population studied, the prevalence of LTV was different among breeds. This supports initial data on the heredity of LTV and the diverse occurrence of LTV among breeds. Our results indicate that LTV type 2 and type 3 are associated with mild and severe CHD development. Therefore, this study has potentially identified an additional risk factor for the development of hip dysplasia.


Preliminary study of quinine pharmacokinetics in pregnant women with malaria-HIV co-infection.
Pregnant women bear the greatest burden of malaria-human immunodeficiency virus co-infection. Previous studies suggest that interaction with antiretroviral drugs may compromise antimalarial pharmacokinetics and treatment outcomes. We conducted a preliminary clinical study to assess quinine pharmacokinetics in Malian pregnant women with acute malaria who reported taking nevirapine-based antiretroviral therapy. Of seven women, six had stable concentrations of nevirapine in the plasma and one had none. Quinine concentrations were lower, and its metabolite 3-hydroxyquinine higher, in the six women with nevirapine than in the one without, and quinine concentrations were below the recommended therapeutic range in 50% of the women. This preliminary observation warrants further research to understand the impact of long-term antiretroviral therapy on the treatment of acute malaria.


Comprehensive Physiologically Based Pharmacokinetic Model to Assess Drug-Drug Interactions of Phenytoin.
Regulatory agencies worldwide expect that clinical pharmacokinetic drug-drug interactions (DDIs) between an investigational new drug and other drugs should be conducted during drug development as part of an adequate assessment of the drug's safety and efficacy. However, it is neither time nor cost efficient to test all possible DDI scenarios clinically. Phenytoin is classified by the Food and Drug Administration as a strong clinical index inducer of CYP3A4, and a moderate sensitive substrate of CYP2C9. A physiologically based pharmacokinetic (PBPK) platform model was developed using GastroPlus® to assess DDIs with phenytoin acting as the victim (CYP2C9, CYP2C19) or perpetrator (CYP3A4). Pharmacokinetic data were obtained from 15 different studies in healthy subjects. The PBPK model of phenytoin explains the contribution of CYP2C9 and CYP2C19 to the formation of 5-(4'-hydroxyphenyl)-5-phenylhydantoin. Furthermore, it accurately recapitulated phenytoin exposure after single and multiple intravenous and oral doses/formulations ranging from 248 to 900 mg, the dose-dependent nonlinearity and the magnitude of the effect of food on phenytoin pharmacokinetics. Once developed and verified, the model was used to characterize and predict phenytoin DDIs with fluconazole, omeprazole and itraconazole, i.e., simulated/observed DDI AUC ratio ranging from 0.89 to 1.25. This study supports the utility of the PBPK approach in informing drug development.


Free Open Access Med(ical edu)cation for critical care practitioners.
Free Open Access Med(ical edu)cation refers to an online community of knowledge relating to medicine. Originating from practitioners in emergency medicine, it has since spread to critical care, internal medicine, prehospital medicine, paediatrics, and allied health professionals and continues to grow at an advanced rate. Weblogs ('blog' for short), emails, social media (in particular Twitter), recorded audio material ((podcasts), and video material are all produced on a daily basis and contribute to the continual professional development of trainees and consultants worldwide. In this article, we explain its background, rise to prominence, and explore some of its controversies.


Analysis of genomic aberrations and gene expression profiling identifies novel lesions and pathways in myeloproliferative neoplasms.
Polycythemia vera (PV), essential thrombocythemia and primary myelofibrosis, are myeloproliferative neoplasms (MPNs) with distinct clinical features and are associated with the JAK2V617F mutation. To identify genomic anomalies involved in the pathogenesis of these disorders, we profiled 87 MPN patients using Affymetrix 250K single-nucleotide polymorphism (SNP) arrays. Aberrations affecting chr9 were the most frequently observed and included 9pLOH (n=16), trisomy 9 (n=6) and amplifications of 9p13.3-23.3 (n=1), 9q33.1-34.13 (n=1) and 9q34.13 (n=6). Patients with trisomy 9 were associated with elevated JAK2V617F mutant allele burden, suggesting that gain of chr9 represents an alternative mechanism for increasing JAK2V617F dosage. Gene expression profiling of patients with and without chr9 abnormalities (+9, 9pLOH), identified genes potentially involved in disease pathogenesis including JAK2, STAT5B and MAPK14. We also observed recurrent gains of 1p36.31-36.33 (n=6), 17q21.2-q21.31 (n=5) and 17q25.1-25.3 (n=5) and deletions affecting 18p11.31-11.32 (n=8). Combined SNP and gene expression analysis identified aberrations affecting components of a non-canonical PRC2 complex (EZH1, SUZ12 and JARID2) and genes comprising a 'HSC signature' (MLLT3, SMARCA2 and PBX1). We show that NFIB, which is amplified in 7/87 MPN patients and upregulated in PV CD34+ cells, protects cells from apoptosis induced by cytokine withdrawal.


[Staging gastritis with the OLGA system: prevalence of advanced stages of gastric atrophy in Mexican patients].
Gastric adenocarcinoma of intestinal type is preceded by inflammation, which produces mucosal atrophy and intestinal metaplasia, progressing eventually to dysplasia and invasive cancer. Recently an international group, the Operative Link on Gastritis Assessment (OLGA) proponed a staging system for gastric biopsies. To recognize the distribution of advanced stages of gastric mucosal atrophy in Mexican patients with dyspepsia according to the OLGA system. We apply the OLGA system for cancer risk (Stages 0 to IV) to 322 gastric biopsies from consecutive patients with dyspepsia. Using the Sydney protocol, we recorded the presence of atrophy, dysplasia and the relationship with ulcer disease. We report the stage of atrophy for each region and the Helicobacter pylori infection status. We documented 72 (22.4%) cases with atrophy, 50 of them (69.4%) were metaplastic-type. Overall, nine biopsies (2.78%) were stage III (all of them with metaplastic-type atrophy) and there was not stage IV cases. We did not find high-grade dysplasia or intramucosal carcinoma. In 8 of subjects with stage III, we observed low-grade dysplasia. We documented gastric ulcer in 5 patients with stage II, 60% of them with associated low-grade dysplasia. Five patients with duodenal ulcer were found in stages 0 and I. We found low prevalence of advanced stages of mucosal gastric atrophy among patients with dyspepsia. However we recognized 9 patients with stage III according to OLGA system worthy of follow-up because the high risk for developing gastric cancer.


A Novel Technique of No-Touch Endoscopic Saphenous Vein Harvesting.
No-touch saphenous vein harvesting is considered an ideal procedure for harvesting good-quality veins, whereas endoscopic vein harvesting (EVH) is considered ideal for decreasing wound complications. The no-touch EVH (NT-EVH) technique, which was developed in 2018, is one of the best vein-harvesting procedures. We have improved this method in several aspects since our initial experience. Herein, we present the details of this technique.


Methylation of the PTENP1 pseudogene as potential epigenetic marker of age-related changes in human endometrium.
The processed pseudogene PTENP1 is involved in the regulation of the expression of the PTEN and acts as a tumor suppressor in many types of malignances. In our previous study we showed that PTENP1 methylation is present not only in tumor, but also in normal endometrium tissues of women over 45 years old. Here we used methylation-specific PCR to analyze methylation status of CpG island located near promoter region of PTENP1 in malignant and non-malignant endometrium tissues collected from 236 women of different age groups. To confirm our results, we also analyzed RNA sequencing and microarray data from 431 women with endometrial cancer from TCGA database. We demonstrated that methylation of PTENP1 is significantly increased in older patients. We also found an age-dependent increase in the level of PTENP1 expression in endometrial tissue. According to our data, PTENP1 methylation elevates the level of the pseudogene sense transcript. In turn, a high level of this transcript correlates with a more favorable prognosis in endometrial cancer. The data obtained suggested that PTENP1 methylation is associated with age-related changes in normal and hyperplastic endometrial tissues. We assumed that age-related increase in PTENP1 methylation and subsequent elevation of its expression may serve as a protective mechanism aimed to prevent malignant transformation of endometrial tissue in women during the perimenopause, menopause, and postmenopause periods.


Perovskites beyond photovoltaics: field emission from morphology-tailored nanostructured methylammonium lead triiodide.
Herein, methylammonium lead triiodide (CH3NH3PbI3) nanorods and nanocrystals were prepared by a facile room-temperature wet chemical method via simple variation of the synthesis parameters. Proper phase formation was confirmed by X-ray diffraction studies, whereas the morphological features were investigated using field emission scanning electron microscopy and transmission electron microscopy. The bonding information and the presence of organic functional groups within the synthesized nanostructures were confirmed using Fourier transform infrared spectroscopy. The work function of the material was calculated using first principles studies. In an attempt to explore the potential of the perovskites beyond photovoltaic applications, the field emission performance of the nanostructured CH3NH3PbI3 was investigated. The turn-on field (electric field corresponding to a 10 μA cm-2 current density) was obtained as 4.2 V μm-1 with the current density reaching up to 96 μA cm-2 for an inter-electrode spacing of 200 μm for the nanorod samples. The emission current stability was tested to be good enough for as long as 2 hours. Finally, finite element-based simulations were performed using ANSYS to obtain a theoretical perception of our experimental findings.


In vitro and in vivo anti-tumoral effect of curcumin against melanoma cells.
Curcumin, the active ingredient from the spice turmeric (Curcuma longa Linn), is known to be an anti-oxidant and an anti-inflammatory agent. It has been demonstrated recently to possess anti-angiogenic effects and pro-apoptotic activities against Ehrlich ascites tumor cells. In the current study, curcumin was found to be cytotoxic in vitro for B16-R melanoma cells resistant to doxorubicin either cultivated as monolayers or grown in three-dimensional (3-D) cultures (spheroids). We have demonstrated that the cytotoxic effect observed in the 2 culture types can be related to the induction of programmed cell death. In our in vivo studies, we examined the effectiveness of a prophylactic immune preparation of soluble proteins from B16-R cells, or a treatment with curcumin as soon as tumoral appearance, alone or in combination, on the murine melanoma B16-R. The combination treatment resulted in substantial inhibition of growth of B16-R melanoma, whereas each treatment by itself showed little effect. Moreover, animals receiving the combination therapy exhibited an enhancement of their humoral anti-soluble B16-R protein immune response and a significant increase in their median survival time (> 82.8% vs. 48.6% and 45.7% respectively for the immunized group and the curcumin-treated group). Our study shows that curcumin may provide a valuable tool for the development of a therapeutic combination against the melanoma.


The interplay between teamwork, clinicians' emotional exhaustion, and clinician-rated patient safety: a longitudinal study.
Effectively managing patient safety and clinicians' emotional exhaustion are important goals of healthcare organizations. Previous cross-sectional studies showed that teamwork is associated with both. However, causal relationships between all three constructs have not yet been investigated. Moreover, the role of different dimensions of teamwork in relation to emotional exhaustion and patient safety is unclear. The current study focused on the long-term development of teamwork, emotional exhaustion, and patient safety in interprofessional intensive care teams by exploring causal relationships between these constructs. A secondary objective was to disentangle the effects of interpersonal and cognitive-behavioral teamwork. We employed a longitudinal study design. Participants were 2100 nurses and physicians working in 55 intensive care units. They answered an online questionnaire on interpersonal and cognitive-behavioral aspects of teamwork, emotional exhaustion, and patient safety at three time points with a 3-month lag. Data were analyzed with cross-lagged structural equation modeling. We controlled for professional role. Analyses showed that emotional exhaustion had a lagged effect on interpersonal teamwork. Furthermore, interpersonal and cognitive-behavioral teamwork mutually influenced each other. Finally, cognitive-behavioral teamwork predicted clinician-rated patient safety. The current study shows that the interrelations between teamwork, clinician burnout, and clinician-rated patient safety unfold over time. Interpersonal and cognitive-behavioral teamwork play specific roles in a process leading from clinician emotional exhaustion to decreased clinician-rated patient safety. Emotionally exhausted clinicians are less able to engage in positive interpersonal teamwork, which might set in motion a vicious cycle: negative interpersonal team interactions negatively affect cognitive-behavioral teamwork and vice versa. Ultimately, ineffective cognitive-behavioral teamwork negatively impacts clinician-rated patient safety. Thus, reducing clinician emotional exhaustion is an important prerequisite of managing teamwork and patient safety. From a practical point of view, team-based interventions targeting patient safety are less likely to be effective when clinicians are emotionally exhausted.


AggNet: Deep Learning From Crowds for Mitosis Detection in Breast Cancer Histology Images.
The lack of publicly available ground-truth data has been identified as the major challenge for transferring recent developments in deep learning to the biomedical imaging domain. Though crowdsourcing has enabled annotation of large scale databases for real world images, its application for biomedical purposes requires a deeper understanding and hence, more precise definition of the actual annotation task. The fact that expert tasks are being outsourced to non-expert users may lead to noisy annotations introducing disagreement between users. Despite being a valuable resource for learning annotation models from crowdsourcing, conventional machine-learning methods may have difficulties dealing with noisy annotations during training. In this manuscript, we present a new concept for learning from crowds that handle data aggregation directly as part of the learning process of the convolutional neural network (CNN) via additional crowdsourcing layer (AggNet). Besides, we present an experimental study on learning from crowds designed to answer the following questions. 1) Can deep CNN be trained with data collected from crowdsourcing? 2) How to adapt the CNN to train on multiple types of annotation datasets (ground truth and crowd-based)? 3) How does the choice of annotation and aggregation affect the accuracy? Our experimental setup involved Annot8, a self-implemented web-platform based on Crowdflower API realizing image annotation tasks for a publicly available biomedical image database. Our results give valuable insights into the functionality of deep CNN learning from crowd annotations and prove the necessity of data aggregation integration.


Distinct effect of contraction and ion transport on NADH fluorescence and lactate production in uterine smooth muscle.
The present studies were designed to monitor metabolic perturbations following changes in contractile activity and ion transport in rat and rabbit myometrium by simultaneous measurement of NADH fluorescence, lactate production and isometric force. Stimulation (127 mM K+) and inhibition (Ca2+-deficient solution) of isometric force development induced a decrease/increase of fluorescence intensity corresponding to a more oxidized/reduced state of tissue pyridine nucleotides, respectively. If tension development was abolished by EGTA, fluorescence changes due to altered ion transport could be monitored. Slow progressive stimulation of the sodium pump by Ca2++-deficient solution resulted in an ouabain (10(-3) M) sensitive monophasic NADH oxidation, which was reversed by inhibition of the pump by isotonic K+ solution. If, however, stimulation of the pump was rapid and maximal (addition of 30 mM KCl to Na+-loaded tissues) the fluorescence response was triphasic and ouabain sensitive: it consisted of an initial NAD reduction followed by a transient NADH oxidation and a second slow NAD reduction. These fluorescence changes are interpreted in terms of separate redox changes in the cytoplasm and mitochondria. The ouabain sensitive stimulation or inhibition of Na/K transport was always accompanied by parallel changes in uterine lactate production irrespective of the actual contractile state of the myometrium. The present results show that 1 fluorescence technique can be applied to monitor metabolic perturbations in the uterus evoked by changes in contractility or ion transport; and 2 aerobic glycolysis and Na/K transport are tightly coupled in the myometrium.


Characterization of 3': 5' -cyclic AMP phosphodiesterase in Klebsiella aerogenes and its role in substrate-accelerated death.
Cyclic AMP phosphodiesterase in Klebsiella aerogenes is a soluble cytoplasmic enzyme with an apparent Km of 0.9 mM and a pH optimum of 7.0. It was inhibited by EDTA, Mg2+ and other metal ions. The enzyme activity was inhibited or activated by some nucleotides but not by any metabolite except pyruvate. It was inhibited by the methylxanthines, caffeine, theophylline and methylisobutylxanthine. During starvation or substrate-accelerated death, the enzyme activity remained essentially constant. It is postulated that during substrate-accelerated death the enzyme acts as a drain on the cellular cyclic AMP levels. The cyclic nucleotide concentrations during substrate-accelerated death are proposed to be controlled directly by adenylate cyclase.


Validation of a newly developed hexaplex real-time PCR assay for screening for presence of GMOs in food, feed and seed.
For years, an increasing number and diversity of genetically modified plants has been grown on a commercial scale. The need for detection and identification of these genetically modified organisms (GMOs) calls for broad and at the same time flexible high throughput testing methods. Here we describe the development and validation of a hexaplex real-time polymerase chain reaction (PCR) screening assay covering more than 100 approved GMOs containing at least one of the GMO targets of the assay. The assay comprises detection systems for Cauliflower Mosaic Virus 35S promoter, Agrobacterium tumefaciens NOS terminator, Figwort Mosaic Virus 34S promoter and two construct-specific sequences present in novel genetically modified soybean and maize that lack common screening elements. Additionally a detection system for an internal positive control (IPC) indicating the presence or absence of PCR inhibiting substances was included. The six real-time PCR systems were allocated to five detection channels showing no significant crosstalk between the detection channels. As part of an extensive validation, a limit of detection (LOD(abs)) < or = ten target copies was proven in hexaplex format. A sensitivity < or = ten target copies of each GMO detection system was still shown in highly asymmetric target situations in the presence of 1,000 copies of all other GMO targets of each detection channel. Furthermore, the applicability to a broad sample spectrum and reliable indication of inhibition by the IPC system was demonstrated. The presented hexaplex assay offers sensitive and reliable detection of GMOs in processed and unprocessed food, feed and seed samples with high efficiency.


Inhibition of interferon-beta responses in multiple sclerosis immune cells associated with high-dose statins.
To determine whether statins affect type 1 interferon responses in relapsing-remitting multiple sclerosis (RRMS). Study effects of atorvastatin on type 1 interferon responses in Jurkat cells, mononuclear cells (MNCs) from therapy-naive patients with RRMS in vitro, and MNCs from interferon-treated RRMS patients in vivo in 4 conditions: no drug, statin only, interferon-beta only, and statin added on to interferon-beta therapy. The study examined clinically stable patients with RRMS: 21 therapy-naive patients and 14 patients receiving interferon-beta with a statin. Statin effects on in vitro and in vivo interferon-beta-induced STAT1 transcription factor activation, expression of interferon-stimulated proteins in MNCs, and serum type 1 interferon activity. In vitro, atorvastatin dose dependently inhibited expression of interferon-stimulated P-Y-STAT1 by 44% (P < .001), interferon regulatory factor 1 protein by 30% (P=.006), and myxovirus resistance 1 protein by 32% (P=.004) compared with no-statin control in MNCs from therapy-naive RRMS patients. In vivo, 9 of 10 patients who received high-dose statins (80 mg) had a significant reduction in interferon-beta therapy-induced serum interferon-α/β activity, whereas only 2 of 4 patients who received medium- dose statins (40 mg) had reductions. High-dose add-on statin therapy significantly blocked interferon-beta function, with less P-Y-STAT1 transcription factor activation, and reduced myxovirus resistance 1 protein and viperin protein production. Medium doses of statins did not change STAT1 activation. High-dose add-on statin therapy significantly reduces interferon-beta function and type 1 interferon responses in RRMS patients. These data provide a putative mechanism for how statins could counteract the beneficial effects of interferon-beta and worsen disease.


Immunoregulatory role of ocular macrophages: the macrophages produce RANTES to suppress experimental autoimmune uveitis.
Murine experimental autoimmune uveitis (EAU) is a model of human uveitis. Ocular-infiltrating macrophages play a crucial role in the generation of tissue damage in EAU. In fact, several chemokines are actually produced in the inflamed eye. The aim of this study was to elucidate the role of ocular macrophage-derived chemokines in EAU. C57BL/6 mice were immunized with human interphotoreceptor retinoid binding protein peptide 1-20, and the EAU severity was scored at multiple time points based on microscopic fundus observations (retinal vascular dilatation and exudates) and histological examinations. The peak inflammatory response was observed 1 wk (day 16) after the beginning of macrophage infiltration to the eye (day 9). Ocular-infiltrating cells were enriched or depleted of macrophages by magnetic beads and analyzed by real-time RT-PCR for chemokine mRNA production. We found that only the macrophage-enriched cells from the eye produced RANTES, and thus proposed that macrophage-derived RANTES facilitated the ocular inflammations. In contrast to our postulate, neutralization of RANTES by specific Ab in vivo on days 9 and 13 exacerbated EAU. We also found that the ratio of ocular CD4/CD8 T cells was markedly increased after treatment. As a result, RANTES neutralization might exacerbate EAU by modulating the type of T cell subsets recruited to the eye. In conclusion, our data provide insight into the immunoregulatory role of macrophages and RANTES in the pathogenesis of ocular inflammation. Not all macrophage-derived chemokines cause local inflammation, since RANTES produced by ocular macrophages appears to suppress EAU.


Time and predictors of response to tumour necrosis factor-alpha blockers in psoriatic arthritis: an analysis of a longitudinal observational cohort.
To determine predictors and time to response to treatment with TNF-alpha blockers in patients with PsA in a longitudinal observational cohort. We performed a cohort analysis of patients who were followed prospectively in a large PsA clinic. Response to treatment was defined as an improvement of at least 40% in active (tender and/or swollen) and swollen joint count (SJC) and 50% improvement in the Psoriasis Area and Severity Index (PASI) score. Ninety-five patients were included in the analysis. Of the total patients, 72.6 and 77.9% demonstrated 40% improvement in active joint counts at 3 and 12 months, respectively. Also, 80.5 and 87.4% of the patients showed 40% improvement in SJC at 3 and 12 months, respectively. A PASI50 was achieved by 54 and 60.4% after 3 and 12 months of treatment, respectively. Of 17 patients who did not achieve 40% improvement in total SJC at 3 months, 11 (64.7%) responded at 12 months. In multivariate analysis, the number of swollen joints at baseline predicted response of total active joints at 12 months [odds ratio (OR) 1.34; P = 0.02], whereas past use of TNF-alpha blocker decreased odds of response (OR 0.05; P = 0.01). TNF-alpha blockers are effective in most PsA patients with the majority responding within 3 months of treatment. A significant proportion of the early non-responders will have a delayed response to treatment. Higher SJC at baseline and no prior use of TNF-alpha blockers predict response.