Characteristics of the 2f(1)-f(2) distortion product otoacoustic emission in a normal hearing population.
Distortion-product otoacoustic emission (DPOAE) fine structure and component characteristics are reported between 0.75 and 16 kHz in 356 clinically normal hearing human subjects ages 10 to 65 yr. Stimulus tones at 55/40, 65/55, and 75/75 dB SPL were delivered using custom designed drivers and a calibration method that compensated for the depth of insertion of the otoacoustic emission (OAE) probe in the ear canal. DPOAE fine structure depth and spacing were found to be consistent with previous reports with depth varying between 3 and 7 dB and average spacing ratios (f/Δf) between 15 and 25 depending on stimulus level and frequency. In general, fine structure depth increased with increasing frequency, likely due to a diminishing difference between DPOAE component levels. Fine structure spacing became wider with increasing age above 8 kHz. DPOAE components were extracted using the inverse fast Fourier transform method, adhering to a strict signal to noise ratio criterion for clearer interpretation. Component data from four age groups between 18 and 55 yr old were available for the stimulus levels of 75/75 dB SPL. The age groups could be differentiated with greater than 90% accuracy when using the level of the component presumed to originate from the DPOAE characteristic frequency place. This accuracy held even for frequencies at and below 4 kHz where the age groups exhibited similar average hearing thresholds.


[Chronic postembolic pulmonary hypertension].
The article deals with chronic pulmonary hypertension consequent upon thromboembolism of the pulmonary artery. From experience gained in the treatment of 15 patients 4 of whom underwent operation, the authors discuss the pathogenesis, clinical picture, diagnosis, and treatment of chronic postembolic pulmonary hypertension.


Squamous Cell Carcinoma and Lymphoma of the Oropharynx: Differentiation Using a Radiomics Approach.
The purpose of this study was to evaluate the diagnostic performance of magnetic resonance (MR) radiomics-based machine learning algorithms in differentiating squamous cell carcinoma (SCC) from lymphoma in the oropharynx. MR images from 87 patients with oropharyngeal SCC (n=68) and lymphoma (n=19) were reviewed retrospectively. Tumors were semi-automatically segmented on contrast-enhanced T1-weighted images registered to T2-weighted images, and radiomic features (n=202) were extracted from contrast-enhanced T1- and T2-weighted images. The radiomics classifier was built using elastic-net regularized generalized linear model analyses with nested five-fold cross-validation. The diagnostic abilities of the radiomics classifier and visual assessment by two head and neck radiologists were evaluated using receiver operating characteristic (ROC) analyses for distinguishing SCC from lymphoma. Nineteen radiomics features were selected at least twice during the five-fold cross-validation. The mean area under the ROC curve (AUC) of the radiomics classifier was 0.750 [95% confidence interval (CI), 0.613-0.887], with a sensitivity of 84.2%, specificity of 60.3%, and an accuracy of 65.5%. Two human readers yielded AUCs of 0.613 (95% CI, 0.467-0.759) and 0.663 (95% CI, 0.531-0.795), respectively. The radiomics-based machine learning model can be useful for differentiating SCC from lymphoma of the oropharynx.


Evaluation of the effect of nimodipine o.d. (extended release) vs nimodipine t.i.d. in the treatment of peripheral vertigo.
Vertigo has a negative impact on quality of life; therefore, it is important to find an effective and convenient therapy that allows patients to continue their everyday tasks as soon as possible and to have a better quality of life. There were two formulations used to assess the effectiveness in vertigo treatment from peripheral origin: nimodipine administrated three times daily (Nimotop®) 30 mg versus nimodipine AP administrated once daily (Tropocer®) 90 mg; both of them in a administrated in a prospective, randomized, double-blind, double dummy, multicenter and parallel-group study, where patients with peripheral vertigo defined as a score ≥7 on the Vertigo-Dizziness Differential Diagnosis Score were included. The patients were evaluated by vertigo severity index and vestibular disability index. In the AP nimodipine group (NAP), vertigo severity index was decreased by 50%: 24% of patients in 14 days, 41% in 4 weeks and 89% in 8 weeks. The vestibular disability index was decreased by 50%: 24% of patients in 15 days, 83% in 4 weeks and 92% of patients in 8 weeks. In the conventional nimodipine group (NC), rate of vertigo severity was decreased by 50%: 17% of patients in 14 days, 41% of patients in 4 weeks and 90% of patients in 8 weeks. The vestibular disability index was decreased by 50%: 15 days in 17% of patients, 53% in 4 weeks and 64% in 8 weeks, without difference between groups. both products were effective and well tolerated in the treatment of peripheral vertigo.


Entry of ketoconazole into Candida albicans.
To define characteristics that determine the entry of ketoconazole (KTZ) into Candida albicans cells, we studied the uptake of [3H]KTZ. The cells rapidly and markedly concentrated the drug: 30% of the final 80-fold intracellular concentration was attained in less than 1 min, and greater than 60% was attained in 10 min. Penetration of [3H]KTZ at an extracellular concentration higher than 0.1875 microM (0.1 microgram/ml) occurred by a simple diffusion mechanism. At lower concentrations, accumulation of the drug was an active, energy-requiring process, dependent at least in part on glycolysis, and pH dependent (optimal pH, 6.6). The active transport system had a high binding affinity (Km = 50 nM) and a high maximum velocity of uptake (Vmax = 1.4 mumol min-1 10(-7) cells). It was not possible to displace intracellular [3H]KTZ with high concentrations of unlabeled KTZ or other antifungal agents. These findings suggest that KTZ is rapidly taken up, highly concentrated, and tightly bound to cellular components of C. albicans.


MAP3K4/CBP-regulated H2B acetylation controls epithelial-mesenchymal transition in trophoblast stem cells.
Epithelial stem cells self-renew while maintaining multipotency, but the dependence of stem cell properties on maintenance of the epithelial phenotype is unclear. We previously showed that trophoblast stem (TS) cells lacking the protein kinase MAP3K4 maintain properties of both stemness and epithelial-mesenchymal transition (EMT). Here, we show that MAP3K4 controls the activity of the histone acetyltransferase CBP, and that acetylation of histones H2A and H2B by CBP is required to maintain the epithelial phenotype. Combined loss of MAP3K4/CBP activity represses expression of epithelial genes and causes TS cells to undergo EMT while maintaining their self-renewal and multipotency properties. The expression profile of MAP3K4-deficient TS cells defines an H2B acetylation-regulated gene signature that closely overlaps with that of human breast cancer cells. Taken together, our data define an epigenetic switch that maintains the epithelial phenotype in TS cells and reveals previously unrecognized genes potentially contributing to breast cancer.


In Situ Understanding of the Effect of Manure on the Availability of Sulfonamide Antibiotics in Soils Using DGT.
In this study, the effects of manure on the availability of sulfonamide antibiotics (SAs) in soils were explored in situ by the Diffusive gradients in thin films (DGT) technique. Five antibiotics, including sulfadiazine (SDZ), sulfamethoxazole (SMX), sulfamethazine (SMZ), sulfachloropyridazine (SCP), and sulfadimethoxine (SDM), were selected as target compounds. Results showed that the manure application to soil could reduce the antibiotic availability indicated by DGT. DGT measurement (CDGT) showed good correlations with the soil solution concentrations (Cd). Manure application can suppress the fluxes of SAs from the soil to the soil solution. Using the DGT-induced soil/sediment flux model (DIFS), the labile pool size (Kdl), the rate constants (k1, k-1) of adsorption and desorption and response time (Tc) of SAs in soils were obtained. The addition of manure increased extractable fraction, labile pool size (Kdl) and k1 but decreased k-1. Together with the nonlinear relationship between DGT fluxes and the reciprocal of diffusive layer thickness (Δg), these findings suggested that the release of SAs from soil particles into the soil solution is thermodynamically and kinetically limited, and the manure application could enhance this limitation. This study offers insight into antibiotic availability in soils caused by manure application.


Soluble mimics of the cytoplasmic face of the human V1-vascular vasopressin receptor bind arrestin2 and calmodulin.
Signal transduction by G protein-coupled receptors (GPCRs) is mediated by interactions between intracellular proteins and exposed motifs on the cytoplasmic face of these receptors. Arrestins bind to GPCRs and modulate receptor function either by interfering with heterotrimeric G protein signaling or by serving as signaling adaptors themselves. Calmodulin interacts with GPCRs triggering a calcium response. We have studied the interaction of arrestin2 and calmodulin with intracellular elements of the human V1-vascular vasopressin receptor (hV1R). For this purpose, we designed, expressed, and purified soluble fusion proteins with the maltose-binding protein (MBP) from Escherichia coli that mimic the intracellular surface of the hV1R. These MBP fusion proteins bind arrestin2 and calmodulin with affinities in the micromolar range. A different series of soluble receptor analogs, named vasopressin receptor 1 elements on a soluble scaffold (V1ROSS) proteins, consist of the third intracellular loop and/or the C-terminal segment of the hV1R receptor juxtaposed on the surface of the MBP. V1ROSS proteins bind calmodulin and a truncated, phosphorylation-independent form of arrestin2 more tightly than the corresponding linear fusion proteins. Thus, embedding receptor loops within the three-dimensional structure of the MBP yields a better representation of the active conformation of these receptor loops than linear receptor peptides fused onto the C terminus of the MBP. V1ROSS proteins provide a valuable tool to study receptor interactions because they are more amenable to structural analysis than the native membrane receptor. These findings set the stage for the detailed structural analysis of these protein-protein interactions that are important for understanding the mechanism of signaling.


Fundus changes following faden operation.
A faden operation was performed in 187 eyes that were studied by indirect binocular ophthalmoscopy at regular intervals after the surgical procedure. Of these eyes, 29 (15.5%) showed ocular fundus abnormalities, with 28 cases of chorioretinal scars at the level of the muscle scleral anchorage and one case of Herpburn-Amalric triangular syndrome of choroidal ischemia. No case of retinal tears was found in this series. Fundus changes are by far the most frequent complication of the faden operation, and to our knowledge, this has not been previously described. The findings of the present study suggest (1) the necessity of routine ophthalmoscopic controls in the postoperative stage regarding those eyes in which the faden operation has been performed, and (2) that the faden operation on the lateral rectus muscle is seriously contraindicated because of the high risk of irreversible macular damage consecutive to these chorioretinal lesions.


Assessing Prevalence and Factors Related to Frailty in Community-Dwelling Older Adults: A Multinomial Logistic Analysis.
Frailty is an age-related clinical condition that typically involves a deterioration in the physiological capacity of various organ systems and heightens the patient's susceptibility to stressors. For this reason, one of the main research goals currently being addressed is that of characterising the impact of frailty in different settings. The main aim of this study is to determine the prevalence of Fried's frailty phenotype among community-dwelling older people and to analyse the factors associated with frailty. In this research study, 582 persons aged 65 years or more participated in this cross-sectional study that was conducted at primary healthcare centres in Málaga, Spain. Sociodemographic, clinical, functional and comprehensive drug therapy data were compiled. The relationship between the independent variables and the different states of frailty was analysed by using a multinomial logistic regression model. Frailty was present in 24.1% of the study sample (95% CI = 20.7-27.6) of whom 54.3% were found to be pre-frail and 21.6% were non-frail. The study variable most strongly associated with frailty was the female gender (OR = 20.54, 95% CI = 9.10-46.3). Other factors found to be associated with the state of frailty included age, dependence for the instrumental activities of daily living (IADL), polymedication, osteoarticular pathology and psychopathology. This study confirms the high prevalence of frailty among community-dwelling older people. Frailty may be associated with many factors. Some of these associated factors may be preventable or modifiable and, thus, provide clinically relevant targets for intervention. This is particularly the case for depressive symptoms, the clinical control of osteoarthritis and the use of polypharmacy.


Dendritic cells,tolerance and therapy of organ allograft rejection.
Donor dendritic cells (DCs) and those of host origin play key roles in the instigation and maintenance of immune responses to organ allografts. In the normal steady state, however, DCs are important for the maintenance of central and peripheral tolerance. Moreover, the presence of those cells in donor hematopoietic cell infusions may facilitate the induction of transplant tolerance. Accrual of information regarding DC tolerogenicity has driven the assessment of DC-based therapy of allograft rejection. Pioneering work demonstrating increased allograft survival after pretransplant infusion of immature donor-derived DC has prompted the evaluation of several approaches to the generation of DCs with tolerogenic/regulatory properties. These include: identification of specific culture conditions for propagation of homogenous populations of immature DCs; pharmacological manipulation of DCs to stabilize their immature/tolerogenic phenotype; and genetic modification of DCs to impair their stimulating ability/enhance their tolerogenicity. These approaches have rendered DCs capable of markedly prolonging experimental allograft (including kidney transplant) survival and promoting donor-specific tolerance. Recently identified molecular signaling pathways that play key roles in the outcome of DC-T cell interaction are likely to become novel targets for manipulation of allograft immunity and for the promotion of transplant tolerance.


Value of urinary albumin-to-creatinine ratio as a predictor of type 2 diabetes in pre-diabetic individuals.
The albumin-to-creatinine ratio (ACR) reflects urinary albumin excretion and is increasingly being accepted as an important clinical outcome predictor. Because of the great public health need for a simple and inexpensive test to identify individuals at high risk for developing type 2 diabetes, it has been suggested that the ACR might serve this purpose. We therefore determined whether the ACR could predict incident diabetes in a well-characterized cohort of pre-diabetic Americans. A total of 3,188 Diabetes Prevention Program (DPP) participants with a mean BMI of 34 kg/m(2) and elevated fasting glucose, impaired glucose tolerance, and baseline urinary albumin excretion measurements were followed for incident diabetes over a mean of 3.2 years. Of the participants, 94% manifested ACR levels below the microalbuminuria range and 21% ultimately developed diabetes during follow-up. Quartiles of ACR (median [range] within quartiles: 1, 3.0 [0.7-3.7]; 2, 4.6 [3.7-5.5]; 3, 7.1 [5.5-9.7]; and 4, 16.5 [9.7-1,578]) were positively associated with age, markers of adiposity and insulin secretion and resistance, blood pressure, and use of antihypertensive agents with antiproteinuric effects and inversely related to male sex and serum creatinine. An elevated hazard rate for developing diabetes with doubling of ACR disappeared after adjustment for covariates. Within the DPP intervention groups (placebo, lifestyle, and metformin), we found no consistent trend in incident diabetes by quartile or decile of ACR. An ACR at levels below the microalbuminuria range does not independently predict incident diabetes in adults at high risk of developing type 2 diabetes.


Optical nonlinearities and enhanced light transmission in soft-matter systems with tunable polarizabilities.
We demonstrate a new class of synthetic colloidal suspensions capable of exhibiting negative polarizabilities, and observe for the first time robust propagation and enhanced transmission of self-trapped light over long distances that would have been otherwise impossible in conventional suspensions with positive polarizabilities. Such light penetration through the strong scattering environment is attributed to the interplay between optical forces and self-activated transparency effects while no thermal effect is involved. By judiciously mixing colloidal particles of both negative and positive polarizabilities, we show that the resulting nonlinear response of these systems can be fine-tuned. Our experimental observations are in agreement with theoretical analysis based on a thermodynamic model that takes into account particle-particle interactions. These results may open up new opportunities in developing soft-matter systems with engineered optical nonlinearities.


Unpacking the species conundrum: philosophy, practice and a way forward.
The history of ecology and evolutionary biology is rife with attempts to define and delimit species. However, there has been confusion between concepts and criteria, which has led to discussion, debate, and conflict, eventually leading to lack of consistency in delimitation. Here, we provide a broad review of species concepts, a clarification of category versus concept, an account of the general lineage concept (GLC), and finally a way forward for species discovery and delimitation. Historically, species were considered as varieties bound together by reproduction. After over 200 years of uncertainty, Mayr attempted to bring coherence to the definition of species through the biological species concept (BSC). This has, however, received much criticism, and the last half century has spawned at least 20 other concepts. A central philosophical problem is that concepts treat species as 'individuals' while the criteria for categorization treats them as 'classes'. While not getting away from this problem entirely, the GLC attempts to provide a framework where lineage divergence is influenced by a number of different factors (and correlated to different traits) which relate to the different species concepts. We also introduce an 'inclusive' probabilistic approach for understanding and delimiting species. Finally, we provide aWallacean (geography related) approach to the Linnaean problem of identifying and delimiting species, particularly for cases of allopatric divergence, and map this to the GLC. Going one step further, we take a morphometric terrain approach to visualizing and understanding differences between lineages. In summary, we argue that while generalized frameworks may work well for concepts of what species are, plurality and 'inclusive' probabilistic approaches may work best for delimitation.


Insulin sensitivity and insulin response in women with gestational diabetes mellitus.
Gestational diabetes mellitus (GDM) is associated with much increased risk of developing diabetes later on in life. Using the frequently sampled intravenous glucose tolerance test and the minimal model analyses we have therefore determined the early insulin response to glucose (EIR) and insulin sensitivity (Si), in women with GDM of different severity (n = 14) and in normal women (n = 10). During the last trimester of pregnancy. GDMs compared to controls had significantly lower EIR (p < 0.001) and Si (p < 0.01). The reduction in EIR was less marked in GDM patients treated with diet alone (n = 6) as compared to GMD patients (n = 8) who subsequently during pregnancy needed treatment also with insulin. The insulin treated GDM group only had higher fasting glucose level than controls (5.2 vs 4.2 mmol/l, p < 0.001). Both GDM subgroups had slightly elevated basal levels of FFA and 3-hydroxybutyrate. Si and EIR were inversely correlated in control women and their fasting glucose correlated both to EIR (r = 0.63, p < 0.05) and to Si (r = 0.59, p < 0.05). In the GDM subgroups Si and EIR were unrelated and there were no correlations between fasting glucose and Si or EIR. These results suggest that glucose intolerance in GDM patients in the last trimester of pregnancy is characterized by both an impaired insulin secretion and an increased resistance to insulin. The impairment of insulin secretion and action increases with the severity of hyperglycemia, and the relative insulin deficiency characterizing GDM patients is associated with a selected defect in insulin action mainly affecting gluco-regulation.


Laminin-332 alters connexin profile, dye coupling and intercellular Ca2+ waves in ciliated tracheal epithelial cells.
Tracheal epithelial cells are anchored to a dynamic basement membrane that contains a variety of extracellular matrix proteins including collagens and laminins. During development, wound repair and disease of the airway epithelium, significant changes in extracellular matrix proteins may directly affect cell migration, differentiation and events mediated by intercellular communication. We hypothesized that alterations in cell matrix, specifically type I collagen and laminin alpha3beta3gamma2 (LM-332) proteins within the matrix, directly affect intercellular communication in ciliated rabbit tracheal epithelial cells (RTEC). Functional coupling of RTEC was monitored by microinjection of the negatively charged fluorescent dyes, Lucifer Yellow and Alexa 350, into ciliated RTEC grown on either a LM-332/collagen or collagen matrix. Coupling of physiologically significant molecules was evaluated by the mechanism and extent of propagated intercellular Ca2+ waves. Expression of connexin (Cx) mRNA and proteins were assayed by reverse transcriptase - polymerase chain reaction and immunocytochemistry, respectively. When compared to RTEC grown on collagen alone, RTEC grown on LM-332/collagen displayed a significant increase in dye transfer. Although mechanical stimulation of RTEC grown on either LM-332/collagen or collagen alone resulted in intercellular Ca2+ waves, the mechanism of transfer was dependent on matrix: RTEC grown on LM-332/collagen propagated Ca2+waves via extracellular purinergic signaling whereas RTEC grown on collagen used gap junctions. Comparison of RTEC grown on collagen or LM-332/collagen matrices revealed a reorganization of Cx26, Cx43 and Cx46 proteins. Alterations in airway basement membrane proteins such as LM-332 can induce connexin reorganizations and result in altered cellular communication mechanisms that could contribute to airway tissue function.


Parenchymal asbestosis due to primary asbestos exposure among ship-breaking workers: report of the first cases from Bangladesh.
We report for the first time asbestosis among ship-breaking workers of Sitakunda in Bangladesh who were exposed to asbestos during ship-based and beach-based operations for at least 10 years. Asbestosis was present among 35% of workers. Years of work (>20) and forced vital capacity (<80% of predicted) were significantly associated with the disease. Currently, global ship-breaking operations are mainly concentrated in the Indian subcontinent, and Bangladesh has the majority share. Ninety per cent of domestic steel is produced in the ship-breaking operations in Bangladesh and is an important contributor to the economy. It also gives employment to more than 100 000 people. It is imperative to medically check up all the workers for benign and malignant diseases causally related to asbestos among these vulnerable population of workers.


Electrically evoked taste threshold.
How accurately can the electrically evoked taste threshold be determined? Why have standards of normal values not yet been generally accepted? In order to answer these questions, 80 normal subjects were examined by means of the method of adjustment. The results showed a greater variability of threshold values and side differences than found by other authors. In explanation, it is suggested that the method of adjustment minimizes the biasing effect of the examiner, which normally leads to an underestimation of the threshold variability as well as of the side differences. The method of adjustment is therefore suitable for certain experimental purposes. For clinical use, a simple procedure like Krarups should be preferred. Due to its limited validity, however, only side differences of 100% or more should be considered pathological. A log-scale with rather large units (25% increment steps) is recommended.


The estimated health impact of alcohol interventions in New Zealand: A modelling study.
To estimate the health impacts of key modelled alcohol interventions among Māori (indigenous peoples) and non-Māori in New Zealand (NZ). Multi-stage life-table intervention modelling study. We modelled two scenarios: (1) business-as-usual (BAU); and (2) an intervention package scenario that included a 50% alcohol tax increase, outlet density reduction from 63 to five outlets per 100 000 people, outlet hours reduction from 112 to 50 per week and a complete ban on all forms of alcohol marketing. The model's population replicates the 2018 NZ population by ethnicity (Māori/non-Māori), age and sex. Alcohol consumption was estimated using nationally representative survey data combined with sales data and corrected for tourist and unrecorded consumption. Disease incidence, prevalence and mortality were calculated using Ministry of Health data. We used dose-response relationships between alcohol and illness from the 2016 Global Burden of Disease study and calculated disability rates for each illness. Changes in consumption were based on the following effect sizes: total intervention package [-30.3%, standard deviation (SD) = 0.02); tax (-7.60%, SD = 0.01); outlet density (-8.64%, SD = 0.01); outlet hours (-9.24%, SD = 0.01); and marketing (-8.98%, SD = 0.02). We measured health gain using health-adjusted life years (HALYs) and life expectancy. Compared with the BAU scenario, the total alcohol intervention package resulted in 726 000 [95% uncertainty interval (UI) = 492 000-913 000] HALYs gained during the life-time of the modelled population. Māori experienced greater HALY gains compared with non-Māori (0.21, 95% UI = 0.14-0.26 and 0.16, 95% UI = 0.11-0.20, respectively). When modelled individually, each alcohol intervention within the intervention package produced similar health gains (~200 000 HALYs per intervention) owing to the similar effect sizes. Modelled interventions for increased alcohol tax, reduced availability of alcohol and a ban on alcohol marketing among Māori and non-Māori in New Zealand (NZ) suggest substantial population-wide health gains and reduced health inequities between Māori and non-Māori.


Synthesis and application of monodisperse oligo(oxyethylene)-grafted polystyrene resins for solid-phase organic synthesis.
In a preliminary investigation by our group, we found that poly(styrene-oxyethylene) graft copolymers (PS-PEG), for example, TentaGel resins, are advantageous for gel-phase (13)C NMR spectroscopy. Because of the solution-like environment provided by the PS-PEG resins, good spectral quality of the attached moiety can be achieved, which is useful for nondestructive on-resin analysis. The general drawbacks of such resins are low loading capacities and the intense signal in the spectra resulting from the PEG linker (>50 units). Here, we describe the characterization of solvent-dependent swelling and reaction kinetics on a new type of resin for solid-phase organic synthesis (SPOS) that allows an accurate monitoring by gel-phase NMR without the above disadvantages. A series of polystyrene-oligo(oxyethylene) graft copolymers containing monodisperse PEG units (n = 2-12) was synthesized. A strong correlation between the linker (PEG) length and the line widths in the (13)C gel-phase spectra was observed, with a grafted PEG chain of 8 units giving similar results in terms of reactivity and gel-phase NMR monitoring to TentaGel resin. Multistep on-resin reaction sequences were performed to prove the applicability of the resins in solid-phase organic synthesis.


Substrate orientation and catalytic specificity in the action of xanthine oxidase: the sequential hydroxylation of hypoxanthine to uric acid.
Xanthine oxidase is a molybdenum-containing enzyme catalyzing the hydroxylation of a sp(2)-hybridized carbon in a broad range of aromatic heterocycles and aldehydes. Crystal structures of the bovine enzyme in complex with the physiological substrate hypoxanthine at 1.8 A resolution and the chemotherapeutic agent 6-mercaptopurine at 2.6 A resolution have been determined, showing in each case two alternate orientations of substrate in the two active sites of the crystallographic asymmetric unit. One orientation is such that it is expected to yield hydroxylation at C-2 of substrate, yielding xanthine. The other suggests hydroxylation at C-8 to give 6,8-dihydroxypurine, a putative product not previously thought to be generated by the enzyme. Kinetic experiments demonstrate that >98% of hypoxanthine is hydroxylated at C-2 rather than C-8, indicating that the second crystallographically observed orientation is significantly less catalytically effective than the former. Theoretical calculations suggest that enzyme selectivity for the C-2 over C-8 of hypoxanthine is largely due to differences in the intrinsic reactivity of the two sites. For the orientation of hypoxanthine with C-2 proximal to the molybdenum center, the disposition of substrate in the active site is such that Arg(880) and Glu(802), previous shown to be catalytically important for the conversion of xanthine to uric acid, play similar roles in hydroxylation at C-2 as at C-8. Contrary to the literature, we find that 6,8-dihydroxypurine is effectively converted to uric acid by xanthine oxidase.


Relevance of social and self-standards in eating disorders.
To compare the importance given to self/other standards by eating disorder (ED) patients and healthy controls. A total of 392 individuals (240 consecutively referred and 152 healthy controls) took part in this study. All subjects were diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision criteria and were female patients. Participants completed the Family Style, Self-Expectations and Emotional related subscales of the Cross-Cultural Questionnaire. Three domains (namely, family standards, self-achievement and physical appearance) were associated with ED. Family standards scores discriminated for the presence of an ED (area under receiver operating characteristic curve equals 0.89), the main predictors being a higher level of importance of physical appearance (p < .001), family standards (p = .029) and conflicts with parents about physical appearance (p < .001). Higher self-standards, in physical appearance, were more relevant in bulimia nervosa and ED not otherwise specified, whereas higher family standards were more associated with anorexia nervosa. High self-standards and social standards are common features in ED. The parallelism that ED may establish between reaching them and their life success may have a crucial role as a developing and maintaining factor in ED.


Principles of microbial PAH-degradation in soil.
Interest in the biodegradation mechanisms and environmental fate of polycyclic aromatic hydrocarbons (PAHs) is motivated by their ubiquitous distribution, their low bioavailability and high persistence in soil, and their potentially deleterious effect on human health. Due to high hydrophobicity and solid-water distribution ratios, PAHs tend to interact with non-aqueous phases and soil organic matter and, as a consequence, become potentially unavailable for microbial degradation since bacteria are known to degrade chemicals only when they are dissolved in water. As the aqueous solubility of PAHs decreases almost logarithmically with increasing molecular mass, high-molecular weight PAHs ranging in size from five to seven rings are of special environmental concern. Whereas several reviews have focussed on metabolic and ecological aspects of PAH degradation, this review discusses the microbial PAH-degradation with special emphasis on both biological and physico-chemical factors influencing the biodegradation of poorly available PAHs.


Long-term effect of Bordetella bronchiseptica infection in neonatal pigs.
A total of 62 gnotobiotic pigs were inoculated with Bordetella bronchiseptica at 7 days of age. There were 25 noninoculated controls. Thirteen of the inoculated pigs were killed and 6 pigs which died were examined between 3 and 5 weeks after inoculations were done (group A). Of these 19 pigs, 16 had severe pneumonia and B bronchiseptica was isolated from the lungs of 17. The turbinates in most of the pigs appeared to have some atrophy. Four other pigs died of pneumonia after they were transferred to the floor in isolation rooms, and 9 inoculated pigs were examined at necropsy at about 112 days of age (group B). Bordetella bronchiseptica was isolated from 8 of the latter 9 pigs, and 7 of the pigs had pneumonic lesions. The turbinates all appeared normal. Thirty inoculated pigs (group C) were killed at 180 days of age. Of these, 22 had pneumonic lesions. Bordetella bronchiseptica was reisolated from the turbinates of 2 of 30 pigs, although the turbinates in all appeared normal. The noninoculated controls examined at necropsy at 3 to 5 weeks, 112 days, and 180 days of age did not have any pulmonic lesions, and their turbinates appeared normal. The inoculated pigs killed at 112 days of age gained an average of 0.25 kg/day, and the control pigs gained 0.31 kg. At 180 days of age, the inoculated pigs had gained 0.41 kg/day and the controls, 0.49 kg. Bordetella bronchiseptica infection in susceptible pigs can cause a severe pneumonia which can persist and reduce the growth rate.


Potential of Mean Force Calculations of Solute Permeation Across UT-B and AQP1: A Comparison between Molecular Dynamics and 3D-RISM.
Membrane channels facilitate the efficient and selective flux of various solutes across biological membranes. A common approach to investigate the selectivity of a channel has been the calculation of potentials of mean force (PMFs) for solute permeation across the pore. PMFs have been frequently computed from molecular dynamics (MD) simulations, yet the three-dimensional reference interaction site model (3D-RISM) has been suggested as a computationally efficient alternative to MD. Whether the two methods yield comparable PMFs for solute permeation has remained unclear. In this study, we calculated potentials of mean force for water, ammonia, urea, molecular oxygen, and methanol across the urea transporter B (UT-B) and aquaporin-1 (AQP1), using 3D-RISM, as well as using MD simulations and umbrella sampling. To allow direct comparison between the PMFs from 3D-RISM and MD, we ensure that all PMFs refer to a well-defined reference area in the bulk or, equivalently, to a well-defined density of channels in the membrane. For PMFs of water permeation, we found reasonable agreement between the two methods, with differences of ≲3 kJ mol-1. In contrast, we found stark discrepancies for the PMFs for all other solutes. Additional calculations confirm that discrepancies between MD and 3D-RISM are not explained by the choice for the closure relation, the definition the reaction coordinate (center of mass-based versus atomic site-based), details of the molecule force field, or fluctuations of the protein. Comparison of the PMFs suggests that 3D-RISM may underestimate effects from hydrophobic solute-channel interactions, thereby, for instance, missing the urea binding sites in UT-B. Furthermore, we speculate that the orientational averages inherent to 3D-RISM might lead to discrepancies in the narrow channel lumen. These findings suggest that current 3D-RISM solvers provide reasonable estimates for the PMF for water permeation, but that they are not suitable to study the selectivity of membrane channels with respect to uncharged nonwater solutes.


Asymmetric bifurcated halogen bonds.
Halogen bonding (XB) is being extensively explored for its potential use in advanced materials and drug design. Despite significant progress in describing this interaction by theoretical and experimental methods, the chemical nature remains somewhat elusive, and it seems to vary with the selected system. In this work we present a detailed DFT analysis of three-center asymmetric halogen bond (XB) formed between dihalogen molecules and variously 4-substituted 1,2-dimethoxybenzene. The energy decomposition, orbital, and electron density analyses suggest that the contribution of electrostatic stabilization is comparable with that of non-electrostatic factors. Both terms increase parallel with increasing negative charge of the electron donor molecule in our model systems. Depending on the orientation of the dihalogen molecules, this bifurcated interaction may be classified as 'σ-hole - lone pair' or 'σ-hole - π' halogen bonds. Arrangement of the XB investigated here deviates significantly from a recent IUPAC definition of XB and, in analogy to the hydrogen bonding, the term bifurcated halogen bond (BXB) seems to be appropriate for this type of interaction.


Time-domain analysis of periodic anisotropic media at oblique incidence: an efficient FDTD implementation.
We describe an efficient implementation of the finite-difference time-domain (FDTD) method as applied to lightwave propagation through periodic media with arbitrary anisotropy (birefringence). A permittivity tensor with non-diagonal elements is successfully integrated here with periodic boundary conditions, bounded computation space, and the split-field update technique. This enables modeling of periodic structures using only one period even with obliquely incident light in combination with both monochromatic (sinusoidal) and wideband (time-domain pulse) sources. Comparisons with results from other techniques in four validation cases are presented and excellent agreement is obtained. Our implementation is freely available on the Web.


Small molecule activation of metabolic enzyme pyruvate kinase muscle isozyme 2, PKM2, circumvents photoreceptor apoptosis.
Photoreceptor cell death is the ultimate cause of vision loss in many retinal disorders, and there is an unmet need for neuroprotective modalities to improve photoreceptor survival. Similar to cancer cells, photoreceptors maintain pyruvate kinase muscle isoform 2 (PKM2) expression, which is a critical regulator in aerobic glycolysis. Unlike PKM1, which has constitutively high catalytic activity, PKM2 is under complex regulation. Recently, we demonstrated that genetically reprogramming photoreceptor metabolism via PKM2-to-PKM1 substitution is a promising neuroprotective strategy. Here, we explored the neuroprotective effects of pharmacologically activating PKM2 via ML-265, a small molecule activator of PKM2, during acute outer retinal stress. We found that ML-265 increased PKM2 activity in 661 W cells and in vivo in rat eyes without affecting the expression of genes involved in glucose metabolism. ML-265 treatment did, however, alter metabolic intermediates of glucose metabolism and those necessary for biosynthesis in cultured cells. Long-term exposure to ML-265 did not result in decreased photoreceptor function or survival under baseline conditions. Notably, though, ML-265-treatment did reduce entrance into the apoptotic cascade in in vitro and in vivo models of outer retinal stress. These data suggest that reprogramming metabolism via activation of PKM2 is a novel, and promising, therapeutic strategy for photoreceptor neuroprotection.


Evaluation of current surgeon practice for patients undergoing lumbar spinal fusion surgery in the United Kingdom.
To ascertain current surgeon practice in the United Kingdom National Health Service for the management of patients undergoing lumbar spinal fusion surgery. Descriptive survey methodology utilised an online questionnaire administered through SurveyMonkey. Eligible participants were all surgeons currently carrying out lumbar spinal fusion surgery in the National Health Service. Two previous surveys and a recent systematic review informed questions. Statistical analyses included responder characteristics and pre-planned descriptive analyses. Open question data were interpreted using thematic analysis. The response rate was 73.8%. Most surgeons (84%) were orthopaedic surgeons. Range of surgeon experience (1-15 years), number of operations performed in the previous 12 mo (4-250), and range of information used to predict outcome was broad. There was some consistency of practice: most patients were seen preoperatively; all surgeons ensured patients are mobile within 3 d of surgery; and there was agreement for the value of post-operative physiotherapy. However, there was considerable variability of practice: variability of protocols, duration of hospital stay, use of discharge criteria, frequency and timing of outpatient follow up, use of written patient information and outcome measures. Much variability was explained through patient-centred care, for example, 62% surgeons tailored functional advice to individual patients. Current United Kingdom surgeon practice for lumbar spinal fusion is described. The surgical procedure and patient population is diverse, and it is therefore understandable that management varies. It is evident that care should be patient-centred. However with high costs and documented patient dissatisfaction it is important that further research evaluates optimal management.


Dexamethasone doses in patients with COVID-19 and hypoxia: A systematic review and meta-analysis.
The optimal dose of dexamethasone for severe/critical COVID-19 is uncertain. We compared higher versus standard doses of dexamethasone in adults with COVID-19 and hypoxia. We searched PubMed and trial registers until 23 June 2023 for randomised clinical trials comparing higher (>6 mg) versus standard doses (6 mg) of dexamethasone in adults with COVID-19 and hypoxia. The primary outcome was mortality at 1 month. Secondary outcomes were mortality closest to 90 days; days alive without life support; and the occurrence of serious adverse events/reactions (SAEs/SARs) closest to 1 month. We assessed the risk of bias using the Cochrane RoB2 tool, risk of random errors using trial sequential analysis, and certainty of evidence using Grading of Recommendations Assessment, Development and Evaluation (GRADE). We included eight trials (2478 participants), of which four (1293 participants) had low risk of bias. Higher doses of dexamethasone probably resulted in little to no difference in mortality at 1 month (relative risk [RR] 0.97, 95% CI: 0.79-1.19), mortality closest to Day 90 (RR 1.01, 95% CI: 0.86-1.20), and SAEs/SARs (RR 1.00, 95% CI: 0.97-1.02). Higher doses of dexamethasone probably increased the number of days alive without invasive mechanical ventilation and circulatory support but had no effect on days alive without renal replacement therapy. Based on low to moderate certainty evidence, higher versus standard doses of dexamethasone probably result in little to no difference in mortality, SAEs/SARs, and days alive without renal replacement therapy, but probably increase the number of days alive without invasive mechanical ventilation and circulatory support.


The Able Worry More? Debt Delinquency, Financial Capability, and Financial Stress.
Research on the link between debt and financial stress is emerging. This study was one of the first attempts to examine the association between debt delinquency and financial stress and the moderating role of financial capability in the association. Delinquencies in three types of debts were examined: (a) mortgage, (b) credit card, and (c) student loan. With data from the 2018 U.S. National Financial Capability Study, multivariate regression results showed that payment delinquencies of mortgage, credit card and student loans were positively, while financial capability was negatively associated with financial stress. Further, surprisingly, the results implied that among consumers with debt delinquencies, financial capability may increase financial stress. If both financial capability's direct and interactive effect were considered, financial capability may decrease financial stress at much smaller rates than those without debt delinquencies. The situation was the worst among consumers with multiple delinquencies, in which the potential net effect of financial capability on financial stress was positive. The results of this study have implications for consumer financial service practices.


Trimerization of monocyanate ester in nanopores.
The effects of nanoconfinement on the reaction kinetics and properties of a monocyanate ester and the resulting cyanurate trimer are studied using differential scanning calorimetry (DSC). On the basis of both dynamic heating scans and isothermal reaction studies, the reaction rate is found to increase with decreasing nanopore size without a change in reaction mechanism. Both the monocyanate ester reactant and cyanurate product show reduced glass transition temperatures (T(g)s) as compared to the bulk; the T(g) depression increases with conversion and is more pronounced for the fully reacted product, suggesting that molecular stiffness influences the magnitude of nanoconfinement effects. Our results are consistent with the accelerated reaction and the T(g) depression found previously for the nanoconfined difunctional cyanate ester, supporting the supposition that intracyclization is not the origin of these effects.


In Vivo and In Vitro Comparison of the DPP-IV Inhibitory Potential of Food Proteins from Different Origins after Gastrointestinal Digestion.
Dipeptidyl-peptidase IV (DPP-IV) plays an essential role in glucose metabolism by inactivating incretins. In this context, food-protein-derived DPP-IV inhibitors are promising glycemic regulators which may act by preventing the onset of type 2 diabetes in personalized nutrition. In this study, the DPP-IV-inhibitory potential of seven proteins from diverse origins was compared for the first time in vitro and in vivo in rat plasma after the intestinal barrier (IB) passage of the indigested proteins. The DPP-IV-inhibitory potentials of bovine hemoglobin, caseins, chicken ovalbumin, fish gelatin, and pea proteins were determined in rat plasma thirty minutes after oral administration. In parallel, these proteins, together with bovine whey and gluten proteins, were digested using the harmonized INFOGEST protocol adapted for proteins. The DPP-IV half-maximal inhibitory concentration (IC50) was determined in situ using Caco-2 cells. The DPP-IV-inhibitory activity was also measured after IB passage using a Caco2/HT29-MTX mixed-cell model. The peptide profiles were analyzed using reversed-phase high-performance liquid chromatography tandem mass spectrometry (RP-HPLC-MS/MS) with MS data bioinformatics management, and the IC50 of the identified peptides was predicted in silico. The in vitro and in vivo DPP-IV-inhibitory activity of the proteins differed according to their origin. Vegetable proteins and hemoglobin yielded the highest DPP-IV-inhibitory activity in vivo. However, no correlation was found between the in vivo and in vitro results. This may be partially explained by the differences between the peptidome analysis and the in silico predictions, as well as the study complexity.


Effects and mechanisms of actions of phytochemicals on Alzheimer's disease neuropathology.
Alzheimer's disease affects millions of people, yet, there are only a limited number approaches for it pharmacological treatment. Thus, identifying factors that decrease the risk of developing Alzheimer's disease is of paramount importance. A growing body of epidemiological and experimental evidence suggests that dietary fruits and vegetables have neuroprotective effects against the harmful effects of oxidative stress, neuroinflammation, and aging. These effects are mediated by various phytochemical compounds found in plants that exhibit antioxidant, anti-inflammatory, and other beneficial properties. This review addresses epidemiological and experimental evidence for the effects and potential mechanisms of several commonly consumed phytochemicals on neuropathology and outcomes of Alzheimer's disease. Based on available evidence, we suggest that regular consumption of bioactive phytochemicals from a variety of fruits and vegetables attenuates age- and insult-related neuropathology in Alzheimer's disease.


Localization and biosynthesis of polyamines in insulin-producing cells.
Two recently developed fluorescence cytochemical methods, specific for spermidine and spermine, were used to localize polyamines in the endocrine pancreas. The polyamines were restricted to the insulin-producing beta-cells and were mainly associated with the secretory granules. Chemical polyamine determinations carried out on isolated rat and mouse pancreatic islets revealed large amounts of polyamines. Compared with extracts of whole pancreas, the islets contained very high concentrations of spermine relative to spermidine. Biosynthesis of polyamines from [3H]ornithine or from [3H]putrescine in isolated islets was significantly stimulated at high glucose concentrations. Moreover, significant incorporation of label from [3H]putrescine was also detected in gamma-aminobutyric acid. This incorporation, however, was not stimulated by high glucose. Possible roles for polyamines associated with the secretory granules in insulin-producing cells are discussed.


Technology, society, and mental illness: challenges and opportunities for assessment and treatment.
Technology is rapidly changing society, and many activities now require the ability to use technology. This situation has the potential to lead to problems for several populations, including the elderly, the disadvantaged, and people with severe mental illness. In this column, we review the state of technology as it affects daily activities. We then review previous efforts to use technology positively for both the assessment and treatment of psychiatric conditions, including posttraumatic stress disorder and severe mental illness. We conclude that technology-based interventions and assessment strategies have the potential to deliver benefit to a wide array of older people and those with severe mental illness, including reaching people who would not have had access otherwise.


Development of an LC-MS-MS method for the quantification of taurine derivatives in marine invertebrates.
Sulfur amino acids, such as taurine, hypotaurine, and thiotaurine, were found in high quantities in tissues of marine symbiotic organisms (e.g., bivalves, tubeworms) living close to hydrothermal vent sites. Therefore, they are assumed to play a key role in the S-oxidizing base metabolism or sulfide detoxification. We propose here a specific, rapid, and original analytical procedure for the direct determination of sulfur amino acids at the level of a few parts per billion in biological samples, avoiding the classical low specific post-column ortho-phthaldialdehyde derivatization step required by non-ultraviolet-absorbing molecules. Indeed, by coupling liquid chromatography on a porous graphitic stationary phase under isocratic conditions (10 mM ammonium acetate buffer adjusted to pH 9.3) to tandem mass spectrometry (ionization process by pneumatically assisted electrospray in negative ion mode), it is possible to perform specific quantification of these metabolites in less than 10 min directly in biological matrices without any derivatization step or other tedious sample treatments. Thus, taurine, hypotaurine, and thiotaurine have been identified and assayed in several deep sea organisms, showing that the developed method is well suited for this kind of application.


Solution NMR structure of the TRIM21 B-box2 and identification of residues involved in its interaction with the RING domain.
Tripartite motif-containing (TRIM) proteins are defined by the sequential arrangement of RING, B-box and coiled-coil domains (RBCC), where the B-box domain is a unique feature of the TRIM protein family. TRIM21 is an E3 ubiquitin-protein ligase implicated in innate immune signaling by acting as an autoantigen and by modifying interferon regulatory factors. Here we report the three-dimensional solution structure of the TRIM21 B-box2 domain by nuclear magnetic resonance (NMR) spectroscopy. The structure of the B-box2 domain, comprising TRIM21 residues 86-130, consists of a short α-helical segment with an N-terminal short β-strand and two anti-parallel β-strands jointly found the core, and adopts a RING-like fold. This ββαβ core largely defines the overall fold of the TRIM21 B-box2 and the coordination of one Zn2+ ion stabilizes the tertiary structure of the protein. Using NMR titration experiments, we have identified an exposed interaction surface, a novel interaction patch where the B-box2 is likely to bind the N-terminal RING domain. Our structure together with comparisons with other TRIM B-box domains jointly reveal how its different surfaces are employed for various modular interactions, and provides extended understanding of how this domain relates to flanking domains in TRIM proteins.


Age and gender-specific reference values of orbital fat and muscle volumes in Caucasians.
To provide age and gender-specific reference values for orbital fat and muscle volumes (MV) in Caucasian adults. Computed tomographic scans of 160 orbits from 52 men and 55 women, aged 20-80 years, not affected by orbital disease were evaluated. Orbital bony cavity volume (OV), fat volume (FV) and MV were calculated by a previously validated method using the software program Mimics. Ratios of FV to OV and of MV to OV were determined. OV, FV and MV were all significantly larger in men than in women (p<0.001), but FV/OV and MV/OV were similar in both sexes. OV and MV did not change with age, but FV increased with advancing age in both women (p<0.001) and men (p<0.001). Linear regression analysis with FV/OV and MV/OV ratios as dependent variables and age and gender as independent variables showed a significant correlation between age and FV/OV (r=0.52, p<0.0001) and age and MV/OV (r=-0.26, p=0.001). Advancing age is associated with an increase of FV/OV and a minor decrease of MV/OV. Gender-specific differences in orbital FV and MV disappear once FV and MV are related to OV, by calculating the ratios FV/OV and MV/OV. Age-specific gender-neutral reference ranges (2.5 and 97.5 percentiles) of FV/OV and MV/OV are presented.


Residential PTSD treatment outcomes during cognitive processing therapy for veterans with and without recent histories of cannabis use.
Prior evidence has suggested that cannabis use is associated with greater posttraumatic stress disorder (PTSD) symptom severity and worse outcomes following trauma-focused treatment. However, lack of high-quality randomized studies necessitates the use of clinical data to clarify the relationship between cannabis use and PTSD treatment to help inform clinical practice. A total of 114 veterans completed cognitive processing therapy in a residential PTSD treatment program. Differences in treatment response between cannabis users and nonusers were evaluated for measures of PTSD, depression, and posttraumatic growth using analysis of covariance to control for pretreatment scores and other drug use. At baseline, cannabis users reported higher levels of PTSD symptom severity relative to nonusers but reported similar levels of depression and posttraumatic growth. Significant differences between groups in the amount of change were not observed on any of the measures from before to after treatment; however, the total sample reported significant improvements in all measures of interest. These results suggest that PTSD treatment outcomes for cannabis users may be similar to nonusers when use is stopped during treatment. Additional data are needed regarding whether outcomes remain similar at follow-up, whether cannabis users maintain abstinence after treatment, and the impact of resumed cannabis use on PTSD symptoms. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Associating quantitative behavioral traits with gene expression in the brain: searching for diamonds in the hay.
Gene expression and phenotypic functionality can best be associated when they are measured quantitatively within the same experiment. The analysis of such a complex experiment is presented, searching for associations between measures of exploratory behavior in mice and gene expression in brain regions. The analysis of such experiments raises several methodological problems. First and foremost, the size of the pool of potential discoveries being screened is enormous yet only few biologically relevant findings are expected, making the problem of multiple testing especially severe. We present solutions based on screening by testing related hypotheses, then testing the hypotheses of interest. In one variant the subset is selected directly, in the other one a tree of hypotheses is tested hierarchical; both variants control the False Discovery Rate (FDR). Other problems in such experiments are in the fact that the level of data aggregation may be different for the quantitative traits (one per animal) and gene expression measurements (pooled across animals); in that the association may not be linear; and in the resolution of interest only few replications exist. We offer solutions to these problems as well. The hierarchical FDR testing strategies presented here can serve beyond the structure of our motivating example study to any complex microarray study. Supplementary data are available at Bioinformatics online.


Total hip arthroplasty in neglected congenital dislocation of the hip.
Between 1984 and 1995, 74 total hip replacements were performed in 64 adult patients who had painful untreated congenital dislocation of the hip. The arthroplasty was performed in the position of the true acetabulum in all patients who had either high or low congenital dislocations of the hip. The femoral head was positioned in the true acetabulum after either osteotomy of the greater trochanter or shortening of the femur, or progressively using external fixation. Information was available on all patients with a followup of 1 to 11 years (mean, 7.2 years). Of the 74 replaced hips, 70 showed marked improvement concerning pain, gait, and mobility, according to the Merle D'Aubigne and Postel scale. Four hips were revised with satisfactory results. The reason for revision was infection in one case and loosening of the plastic cup in three cases. Shortening of the femur by removing a segment of bone below the level of the lesser trochanter followed by osteosynthesis without osteotomy of the greater trochanter was found to be the best method for treating bilateral and several unilateral high congenital dislocation of the hip.


Overlapping group screening for detection of gene-gene interactions: application to gene expression profiles with survival trait.
The development of a disease is a complex process that may result from joint effects of multiple genes. In this article, we propose the overlapping group screening (OGS) approach to determining active genes and gene-gene interactions incorporating prior pathway information. The OGS method is developed to overcome the challenges in genome-wide data analysis that the number of the genes and gene-gene interactions is far greater than the sample size, and the pathways generally overlap with one another. The OGS method is further proposed for patients' survival prediction based on gene expression data. Simulation studies demonstrate that the performance of the OGS approach in identifying the true main and interaction effects is good and the survival prediction accuracy of OGS with the Lasso penalty is better than the ordinary Lasso method. In real data analysis, we identify several significant genes and/or epistasis interactions that are associated with clinical survival outcomes of diffuse large B-cell lymphoma (DLBCL) and non-small-cell lung cancer (NSCLC) by utilizing prior pathway information from the KEGG pathway and the GO biological process databases, respectively. The OGS approach is useful for selecting important genes and epistasis interactions in the ultra-high dimensional feature space. The prediction ability of OGS with the Lasso penalty is better than existing methods. The OGS approach is generally applicable to various types of outcome data (quantitative, qualitative, censored event time data) and regression models (e.g. linear, logistic, and Cox's regression models).


Research Strategies for Orphan G-Protein-Coupled Receptors.
The Society for Medicines Research meeting on orphan G-protein-coupled receptors (GPCRs) was held on March 7, 2002 at the Novartis Research Centre in Horsham, United Kingdom. The dynamic and highly competitive field of GPCR research was the focus of this SMR meeting, featuring speakers from Pharmagene, Manchester University, GlaxoSmithKline, The Royal Danish School of Pharmacy, Pfizer, AstraZeneca, Merck and Synaptic. The meeting attracted a large and enthusiastic audience interested in the research efforts of leading international research teams in the area of GPCR research, whose immediate aim is to evolve the novel molecular targets into lead discovery programs. (c) 2002 Prous Science. All rights reserved.


Foot sole and ankle muscle inputs contribute jointly to human erect posture regulation.
In order to assess the relative contribution and the interactions of the plantar cutaneous and muscle proprioceptive feedback in controlling human erect posture, single or combined vibratory stimuli were applied to the forefoot areas and to the tendons of the tibialis anterior muscles of nine standing subjects using various vibration frequency patterns (ranging from 20 to 80 Hz). The variations in the centre of foot pressure, ankle angle and the EMG activities of the soleus and tibialis anterior muscles of each subject were recorded and analysed. Separate stimulation of the plantar forefoot zones or the tibialis anterior muscles always resulted in whole-body tilts oppositely directed backwards and forwards, respectively, the amplitude of which was proportional to the vibration frequency. EMG activity of ankle muscles also varied according to the direction of the postural responses. However, the same vibration frequency did not elicit equivalent postural responses: in the low frequency range, tactile stimulation induced stronger postural effects than proprioceptive stimulation, and the converse was the case for the higher frequency range. Under sensory conflict conditions, i.e. foot sole-flexor ankle muscle co-stimulation, the direction of the body tilts also varied according to the difference and the absolute levels of the vibration frequencies. In all cases, the resulting postural shifts always corresponded to the theoretical sum of the isolated effects observed upon vibrating each of these two sensory channels. We proposed that tactile and proprioceptive information from the foot soles and flexor ankle muscles might be co-processed following a vector addition mode to subserve the maintenance of erect stance in a complementary way.


In Vitro Studies on Ameliorative Effects of Limonene on Cadmium-Induced Genotoxicity in Cultured Human Peripheral Blood Lymphocytes.
Cadmium (Cd) is a non-redox metal that can indirectly cause oxidative stress by depleting cellular levels of glutathione. It is well-known for the generating reactive oxygen species (ROS) such as hydroxyl radicals, superoxide anions, nitric oxide, and hydrogen peroxide. The latter inactivates antioxidant enzymes and induces lipid peroxidation and DNA damage in cells. In our study, we have investigated the ameliorative effect of limonene against Cd-induced genotoxicity using various biomarkers such as sister chromatid exchange (SCE), comet, and lipid peroxidation (LPA) assays in cultured human peripheral blood lymphocytes (PBL) from healthy individuals. It is a naturally occurring flavonoid, found in essential oil of citrus fruits. Limonene at 20-μM and 100-μM concentrations had significantly (P < 0.05 and P < 0.01) reduced the SCE frequency, tail moment, and peroxidation of lipids. Ameliorative effect of limonene was also determined by measuring the activity of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT). We found a significant increase (P < 0.05) in the enzyme activity after limonene treatment. We also studied the effect of GSTP1 gene polymorphism on Cd-induced genotoxicity and antigenotoxic potential of limonene. We found a significant decrease (P < 0.05) in SCE frequency, tail moment, and lipid peroxidation after limonene treatment compared to Cd. However, we did not observe any significant relationship (P > 0.05) between GSTP1 polymorphism and Cd, limonene genotoxicity and antigenotoxicity, respectively.


[Role of Toll-like receptors in the therapeutic effect of probiotics].
We considered data on the interaction of probiotics with Toll-like receptors. Is was shown that the probiotic effect may have and intact bacterial cells, living or dead, and their structural components. The basis of the surface structures and the DNA of probiotics is the ability to interact with Toll-like receptors, and this interaction can lead to uneven effects, which greatly depends on the strain of probiotic. Hence the obvious need for a more detailed study of the effects of the interaction of probiotic strains with the Toll-receptors. The criteria for evaluating new drugs probiotioticheskih and feasibility of optimal probiotic in a particular clinical case.


In vitro antioxidant and antihyperlipidemic activities of Bauhinia variegata Linn.
To evaluate the ethanolic and aqueous extracts of Bauhinia variegata Linn. for in vitro antioxidant and antihyperlipidemic activity. Ethanolic and aqueous extracts of the stem bark and root of B. variegata Linn. were prepared and assessed for in vitro antioxidant activity by various methods namely total reducing power, scavenging of various free radicals such as 1,2-diphenyl-2-picrylhydrazyl (DPPH), super oxide, nitric oxide, and hydrogen peroxide. The percentage scavenging of various free radicals were compared with standard antioxidants such as ascorbic acid and butylated hydroxyl anisole (BHA). The extracts were also evaluated for antihyperlipidemic activity in Triton WR-1339 (iso-octyl polyoxyethylene phenol)-induced hyperlipidemic albino rats by estimating serum triglyceride, very low density lipids (VLDL), cholesterol, low-density lipids (LDL), and high-density lipid (HDL) levels. Significant antioxidant activity was observed in all the methods, (P < 0.01) for reducing power and (P < 0.001) for scavenging DPPH, super oxide, nitric oxide, and hydrogen peroxide radicals. The extracts showed significant reduction (P < 0.01) in cholesterol at 6 and 24 h and (P < 0.05) at 48 h. There was significant reduction (P < 0.01) in triglyceride level at 6, 24, and 48 h. The VLDL level was also significantly (P < 0.05) reduced from 24 h and maximum reduction (P < 0.01) was seen at 48 h. There was significant increase (P < 0.01) in HDL at 6, 24, and 48 h. From the results, it is evident that alcoholic and aqueous extracts of B. variegata Linn. can effectively decrease plasma cholesterol, triglyceride, LDL, and VLDL and increase plasma HDL levels. In addition, the alcoholic and aqueous extracts have shown significant antioxidant activity. By the virtue of its antioxidant activity, B. variegata Linn. may show antihyperlipidemic activity.


Alloimmunization among transfusion-dependent thalassemia patients.
Thalassemia is a common hemoglobin disorder in Iran and one of the major public health problems. Although blood transfusions are lifesavers for thalassemia patients, they may be associated with some complications especially erythrocyte alloimmunization. The purpose of this study was to investigate the prevalence of red blood cell alloantibodies and to determine types of these antibodies among multiple-transfused thalassemic patients. A total of 313 thalassemia patients in the northeast of Iran, who received regular blood transfusion, were included in this study. Screening of antibodies was performed on fresh serum of all patients and then antibodies were identified in patients' serum that had positive antibody screening test using a panel of recognized blood group antigens. We identified 12 alloantibodies in 9 patients (2.87%) that all were against Rhesus (Rh) blood group antigens (D, C, E). Three patients developed 2 antibodies, and others had one antibody. The most common alloantibodies were Anti-D (88.88%) and followed by Anti-C and Anti-E. Higher frequency of alloimmunization was observed in female, Rh negative and splenectomized patients. This study showed that evaluation of the packed cells for Rh (C, E) from the start of transfusion can be helpful in decreasing the rate of alloantibody synthesis.


HIV prevention trial design in an era of effective pre-exposure prophylaxis.
Pre-exposure prophylaxis (PrEP) has demonstrated remarkable effectiveness protecting at-risk individuals from HIV-1 infection. Despite this record of effectiveness, concerns persist about the diminished protective effect observed in women compared with men and the influence of adherence and risk behaviors on effectiveness in targeted subpopulations. Furthermore, the high prophylactic efficacy of the first PrEP agent, tenofovir disoproxil fumarate/emtricitabine (TDF/FTC), presents challenges for demonstrating the efficacy of new candidates. Trials of new agents would typically require use of non-inferiority (NI) designs in which acceptable efficacy for an experimental agent is determined using pre-defined margins based on the efficacy of the proven active comparator (i.e. TDF/FTC) in placebo-controlled trials. Setting NI margins is a critical step in designing registrational studies. Under- or over-estimation of the margin can call into question the utility of the study in the registration package. The dependence on previous placebo-controlled trials introduces the same issues as external/historical controls. These issues will need to be addressed using trial design features such as re-estimated NI margins, enrichment strategies, run-in periods, crossover between study arms, and adaptive re-estimation of sample sizes. These measures and other innovations can help to ensure that new PrEP agents are made available to the public using stringent standards of evidence.


Regulation of T-cell receptor signalling by membrane microdomains.
There is now considerable evidence suggesting that the plasma membrane of mammalian cells is compartmentalized by functional lipid raft microdomains. These structures are assemblies of specialized lipids and proteins and have been implicated in diverse biological functions. Analysis of their protein content using proteomics and other methods revealed enrichment of signalling proteins, suggesting a role for these domains in intracellular signalling. In T lymphocytes, structure/function experiments and complementary pharmacological studies have shown that raft microdomains control the localization and function of proteins which are components of signalling pathways regulated by the T-cell antigen receptor (TCR). Based on these studies, a model for TCR phosphorylation in lipid rafts is presented. However, despite substantial progress in the field, critical questions remain. For example, it is unclear if membrane rafts represent a homogeneous population and if their structure is modified upon TCR stimulation. In the future, proteomics and the parallel development of complementary analytical methods will undoubtedly contribute in further delineating the role of lipid rafts in signal transduction mechanisms.


Sonographic appearance of hemorrhagic ovarian cyst with acute abdomen by transvaginal scan.
Hemorrhagic ovarian cyst (HOC) which is one of the functional cysts, is often involved in acute abdomen leading to laparatomy intervention. The reason for this mainly lies in the fact that it is easily misdiagnosed as an organic mass because of the presence of lower abdominal pain and the variable appearance of ultrasonographic images at presentation. We analyzed 15 cases of HOC associated with acute abdomen, of which in 2 cases the disease was confirmed by laparotomy. The remaining 13 cases were followed-up clinically and by daily transvaginal sonography (TVS) from the first detection of the cyst until complete resolution. The TVS images showed a variety of changes; however, when the images or their magnified views were observed precisely, important diagnostic characteristics were found which were classified into 3 categories: type 1 images showed mixed hypoechoic and hyperechoic areas, the demarcation line between which appeared as a thin or thick septum-like echo of smooth formation; type 2 images showed hypoechoic background and vertical, horizontal, or lamellar thin or thick thread-like echoes with an overall reticular-like or sponge-like pattern; and type 3 images showed an overall hyperechoic and solid pattern. Type 1 and 2 images occurred more frequent (93.3%), and only 1 case had a type 3 image. In all image types, septum-like or thread-like echoes were seen, TVS type 1 and 2 images showed a clear division into hyperechoic and other areas with the passing of time which was finally changed into a cystic pattern and disappeared. Severe lower abdominal pain was present for 1 to 3 hours in 12 cases (80%), 4 to 6 hours in 2 cases (13.3%), and 11 hours in 1 case (6.7%). Other characteristics of HOC may be its most frequent occurrence in the young age group (10 to 20 years old, 80.0%) and in the luteal phase (84.6%). With operative cases, histopathological diagnosis was HOC. The clinical and particularly TVS findings described in the present study are of significant value in differential diagnosis of HOC with acute abdomen from other disorders presenting with acute abdomen.


Oxidative potential of size-fractionated atmospheric aerosol in urban and rural sites across Europe.
In this study we applied several assays, an in vitro rat alveolar macrophage model, a chemical ROS probe (DTT, dithiothreitol), and cytokine induction (TNFα) to examine relationships between PM-induced generation of reactive oxygen species (ROS) and PM composition, using a unique set of size-resolved PM samples obtained from urban and rural environments across Europe. From April-July 2012, we collected PM from roadside canyon, roadside motorway, and background urban sites in each of six European cities and from three rural sites spanning the continent. A Hi-Vol sampler was used to collect PM in three size classes (PM>7, PM7-3, PM3) and PM was characterized for total elements, and oxidative activity quantified in unfiltered and filtered PM extracts. We measured a remarkable uniformity in air concentrations of ROS and especially DTT activity across the continent. Only a 4-fold difference was documented for DTT across the urban sites and a similar variance was documented for ROS, implying that chemical drivers of oxidative activity are relatively similar between sites. The ROS and DTT specific activity was greater at urban background sites (and also rural sites) than at urban canyon locations. PM3 dominated the size distribution of both ROS activity (86% of total) and DTT activity (76% of total), reflecting both the large contribution of PM3 to total PM mass levels and importantly the higher specific oxidative activity of the PM3 in comparison with the larger particles. The soluble fraction of total activity was very high for DTT (94%) as well as for ROS (64%) in the PM3. However in the larger PM size fractions the contributions of the insoluble components became increasingly significant. The dominance of the insoluble PM drivers of activity was particularly evident in the TNFα data, where the insoluble contribution to cytokine production could be 100-fold greater than that from soluble components. ROS and DTT activity were strongly correlated in the PM3 (r = 0.93), however oxidative activity was not correlated with any measured inorganic element in this size cut. In contrast, significant correlations of both ROS and DTT oxidative activity with specific groups of chemical elements were documented in the larger PM size fractions.


Delivery of oligonucleotides to bone marrow to modulate ferrochelatase splicing in a mouse model of erythropoietic protoporphyria.
Erythropoietic protoporphyria (EPP) is a rare genetic disease in which patients experience acute phototoxic reactions after sunlight exposure. It is caused by a deficiency in ferrochelatase (FECH) in the heme biosynthesis pathway. Most patients exhibit a loss-of-function mutation in trans to an allele bearing a SNP that favors aberrant splicing of transcripts. One viable strategy for EPP is to deploy splice-switching oligonucleotides (SSOs) to increase FECH synthesis, whereby an increase of a few percent would provide therapeutic benefit. However, successful application of SSOs in bone marrow cells is not described. Here, we show that SSOs comprising methoxyethyl-chemistry increase FECH levels in cells. We conjugated one SSO to three prototypical targeting groups and administered them to a mouse model of EPP in order to study their biodistribution, their metabolic stability and their FECH splice-switching ability. The SSOs exhibited distinct distribution profiles, with increased accumulation in liver, kidney, bone marrow and lung. However, they also underwent substantial metabolism, mainly at their linker groups. An SSO bearing a cholesteryl group increased levels of correctly spliced FECH transcript by 80% in the bone marrow. The results provide a promising approach to treat EPP and other disorders originating from splicing dysregulation in the bone marrow.


Investigation of Killer Immunoglobulin-like Receptor (KIR) and HLA Genotypes to Predict the Occurrence of Acute Allograft Rejection after Kidney Transplantation.
After kidney transplantation, natural killer (NK) cells play a pivotal role in triggering the immune response to the allogeneic grafts primarily by their killer-cell immunoglobulin-like receptors (KIR). This process may be one mechanism that contributes to graft rejection. In this study, we have evaluated whether acute rejection after kidney transplantation was associated with predicted NK cell alloreactivity based on KIR gene and ligand along with KIR/HLA compound genotype analysis. After kidney transplantation, natural killer (NK) cells play a pivotal role in triggering the immune response to the allogeneic grafts primarily by their killer-cell immunoglobulin-like receptors (KIR). This process may be one mechanism that contributes to graft rejection. In this study, we have evaluated whether acute rejection after kidney transplantation was associated with predicted NK cell alloreactivity based on KIR gene and ligand along with KIR/HLA compound genotype analysis. DNA from 65 patients with biopsy-proven acute kidney allograft rejection (AKAR), 61 clinically stable graft function (SGF) recipients and 176 healthy subjects were identified for the presence or absence of 10 variable KIR genes (both activating and inhibitory receptors) and their HLA ligands using polymerase chain reaction-sequence specific primers (PCR-SSP) assay. Although no significant difference in the frequency of individual KIR genes, was found the gene content, and the haplotypic distribution between the three categories were detected, the frequency of the KIR3DL1+HLA-Bw4*A allele combination was significantly lower in AKAR patients compared to SGF recipients (p=0.004, OR=0.34, CI=0.16-0.72) and healthy subjects (p=0.019, OR=0.47, CI=0.25-0.89). Kaplan-Meier survival test showed that the KIR3DL1+HLA-Bw4*A allele combination could be considered protective for AKAR (p=0.04 by log-rank). The results of this study suggest that KIR/HLA polymorphism may be a genetic susceptibility factor to alloreactivity dysfunction in the NK cells of patients with AKAR. It is likely that a KIR/HLA combinatorial study can be beneficial in predicting AKAR occurrence for the purpose of selecting donors appropriately.


Involvement of aquaporins in a mouse model of rotavirus diarrhea.
Rotavirus diarrhea is a major worldwide cause of infantile gastroenteritis; however, the mechanism responsible for intestinal fluid loss remains unclear. Water transfer across the intestinal epithelial membrane seems to occur because of aquaporins (AQPs). Accumulating evidence indicates that alterations in AQPs may play an important role in pathogenesis. Here, we focus on changes in AQPs in a mouse model of rotavirus diarrhea. In the present study, 32 of 35 mice developed diarrhea and mild dehydration within 24 hours after infection with rotavirus strain SA11. Intestinal epithelial cells demonstrated cytoplasmic vacuolation, malaligned villi, and atrophy. AQP1 expression was significantly attenuated in the ileum and colon in comparison with controls; likewise, AQP4 and -8 protein expression were significantly decreased in the colon of rotavirus diarrhea-infected mice. In contrast, AQP3 protein expression was significantly increased in the colon of rotavirus-infected mice in comparison with controls. These results indicate that rotavirus diarrhea is associated with the downregulation of AQP1, -4, and -8 expression. Therefore, AQPs play an important role in rotavirus diarrhea.


Identification of potential mRNA biomarkers in peripheral blood lymphocytes for human exposure to ionizing radiation.
Since early in the Atomic Age, biological indicators of radiation exposure have been sought, but currently available methods are not entirely satisfactory. Using cDNA microarray hybridization to discover new potential biomarkers, we have identified genes expressed at increased levels in human peripheral blood lymphocytes after ex vivo irradiation. We recently used this technique to identify a large set of ionizing radiation-responsive genes in a human cell line (Oncogene 18, 3666-3672, 1999). The present set of radiation markers in peripheral blood lymphocytes was identified 24 h after treatment, and while the magnitude of mRNA induction generally decreased over time, many markers were still significantly elevated up to 72 h after irradiation. In all donors, the most highly responsive gene identified was DDB2, which codes for the p48 subunit of XPE, a protein known to play a crucial role in repair of ultraviolet (UV) radiation damage in DNA. Induction of DDB2, CDKN1A (also known at C1P1/WAF1) and XPC showed a linear dose-response relationship between 0.2 and 2 Gy at 24 and 48 h after irradiation, with less linearity at earlier or later times. These results suggest that relative levels of gene expressions in peripheral blood cells may provide estimated of environmental radiation exposures.


Postoperative Meld-Lactate and Isolated Lactate Values As Outcome Predictors Following Orthotopic Liver Transplantation.
The Model for End Stage Liver (MELD) score is validated to predict pretransplant mortality. However, as a predictor of postoperative outcomes, its utility has proven inconsistent. Recently developed MELD-Lactate models better predict 30-day survival as compared with the MELD and MELD-Sodium scores. We compared the MELD-Lactate, original MELD, and MELD-Sodium formulae and the initial postoperative lactate as predictors of 30-day and in-hospital mortality following liver transplantation.Adult patients (n = 989) undergoing orthotopic liver transplant between 2002 to 2013 were included. In addition to the previous models, the first postoperative lactate value and a newly derived Mount Sinai MELD-Lactate score and associated c-statistics were compared.The Mount Sinai MELD-Lactate model yielded the highest c-statistic value (0.749), followed by the original MELD-Lactate (0.740), initial lactate value (0.729), postoperative MELD (0.653), and MELD-Sodium (0.641) models in predicting survival at 30 days following liver transplantation. For in-hospital mortality, the original MELD-Lactate model had slightly higher c-statistic (0.739) compared with the Mount Sinai MELD-Lactate model (0.734). Despite the distribution differences in the MELD-Lactate models, the model validation results, both from cross-validation and bootstrap methods, were similar.Postoperative MELD-Lactate and isolated postoperative lactate values were moderately predictive of 30-day and in-hospital mortality following liver transplantation in this patient cohort.


Sustaining observations of the unsteady ocean circulation.
Sustained observations of ocean properties reveal a global warming trend and rising sea levels. These changes have been documented by traditional ship-based measurements of ocean properties, whereas more recent Argo profiling floats and satellite records permit estimates of ocean changes on a near real-time basis. Through these and newer methods of observing the oceans, scientists are moving from quantifying the 'state of the ocean' to monitoring its variability, and distinguishing the physical processes bringing signals of change. In this paper, I give a brief overview of the UK contributions to the physical oceanographic observations, and the role they have played in the wider global observing systems. While temperature and salinity are the primary measurements of physical oceanography, new transbasin mooring arrays also resolve changes in ocean circulation on daily timescales. Emerging technologies permit routine observations at higher-than-ever spatial resolutions. Following this, I then give a personal perspective on the future of sustained observations. New measurement techniques promise exciting discoveries concerning the role of smaller scales and boundary processes in setting the large-scale ocean circulation and the ocean's role in climate. The challenges now facing the scientific community include sustaining critical observations in the case of funding system changes or shifts in government priorities. These long records will enable a determination of the role and response of the ocean to climate change.


Abdominal compartment syndrome from bleeding duodenal diverticulum.
Duodenal diverticuli are acquired false diverticuli of unknown etiology. Although mostly asymptomatic, they can occasionally cause upper gastrointestinal hemorrhage, rarely with massive bleeding. In this report, we present (to the best of our knowledge) the first reported case of duodenal diverticular bleeding, causing abdominal compartment syndrome. Albeit a rare event, duodenal diverticular bleeding should be included in the differential diagnosis of upper gastrointestinal bleeding. As with our case, a multidisciplinary approach to managing such patients is crucial.


Zymogen activation confers thermodynamic stability on a key peptide bond and protects human cationic trypsin from degradation.
Human cationic trypsinogen, precursor of the digestive enzyme trypsin, can be rapidly degraded to protect the pancreas when pathological conditions threaten, while trypsin itself is impressively resistant to degradation. For either form, degradation is controlled by two necessary initial proteolytic events: cleavage of the Leu81-Glu82 peptide bond by chymotrypsin C (CTRC) and cleavage of the Arg122-Val123 peptide bond by trypsin. Here we demonstrate that the Leu81-Glu82 peptide bond of human cationic trypsin, but not trypsinogen, is thermodynamically stable, such that cleavage by CTRC leads to an equilibrium mixture containing 10% cleaved and 90% uncleaved trypsin. When cleaved trypsin was incubated with CTRC, the Leu81-Glu82 peptide bond was re-synthesized to establish the same equilibrium. The thermodynamic stability of the scissile peptide bond was not dependent on CTRC or Leu-81, as re-synthesis was also accomplished by other proteases acting on mutated cationic trypsin. The Leu81-Glu82 peptide bond is located within a calcium binding loop, and thermodynamic stability of the bond was strictly dependent on calcium and on the calcium-coordinated residue Glu-85. Trypsinolytic cleavage of the Arg122-Val123 site was also delayed in trypsin relative to trypsinogen in a calcium-dependent manner, but for this bond cleavage was modulated by kinetic rather than thermodynamic control. Our results reveal that the trypsinogen to trypsin conformational switch modulates cleavage susceptibility of nick sites by altering both the thermodynamics and kinetics of cleavage to protect human cationic trypsin from premature degradation.


Decreased connexin32 and a characteristic enzyme phenotype in clofibrate-induced preneoplastic lesions not shared with spontaneously occurring lesions in the rat liver.
Two different types of focal preneoplastic lesions, tentatively named Type I and II lesions, were recognized in the liver of rats chronically treated with clofibrate for 104 weeks. Type I lesions were characterized by mostly negative glucose-6-phosphate dehydrogenase (G6PD) activity (6 out of 10, 60%) and positive expression of succinate dehydrogenase (10 out of 10, 100%), in addition to the previously documented complete lack of expression of glutathione S-transferase, placental form (GST-P) and gamma-glutamyl transpeptidase (GGT). Furthermore, most importantly, Type I lesions exhibited a clear decrease in immunohistochemically demonstrated connexin32 (Cx32) spot counts on their hepatocyte membranes, similarly to nitrosamine-induced lesions. In contrast, Type II lesions, mostly small in size and positively expressing GST-P and/or GGT and G6PD, similarly to their previously reported nitrosamine-induced counterparts, did not exhibit a significant decrease in Cx32 count. In addition, spontaneously occurring lesions, again sharing the same enzyme phenotype, did not show a decrease in Cx32. The results indicate that: (i) a clear distinction between the two lesions, with Type I being involved in clofibrate-induced tumors and Type II being more likely to be spontaneous in nature; (ii) a decrease in Cx32 is closely linked to lesion development and possibly stage of progression, irrespective of the enzyme phenotype and the applied carcinogen; (iii) the unaltered condition of Cx32 may suggest a slow growing or non-progressive nature.


Upregulation of SLC7A11/xCT creates a vulnerability to selenocystine-induced cytotoxicity.
The SLC7A11/xCT cystine and glutamate antiporter has emerged as an attractive target for cancer therapy due to its selective overexpression in multiple cancers and its role in preventing ferroptosis. Utilizing pharmacological and genetic approaches in hepatocellular carcinoma cell lines, we demonstrate that overexpression of SLC7A11 engenders hypersensitivity towards L-selenocystine, a naturally-occurring diselenide that bears close structural similarity to L-cystine. We find that abundance of SLC7A11 positively correlates with sensitivity to L-selenocystine, but surprisingly, not to Erastin, an inhibitor of SLC7A11 activity. Our data indicate that SLC7A11 acts as a transport channel for L-selenocystine, which preferentially incites acute oxidative stress and damage eventuating to cell death in cells that highly express SLC7A11. Hence, our findings raise the prospect of L-selenocystine administration as a novel strategy for targeting cancers that upregulate SLC7A11 expression.


Laser flash photolysis generation and kinetic studies of porphyrin-manganese-oxo intermediates. Rate constants for oxidations effected by porphyrin-Mn(V)-oxo species and apparent disproportionation equilibrium constants for porphyrin-Mn(IV)-oxo species.
Porphyrin-manganese(V)-oxo and porphyrin-manganese(IV)-oxo species were produced in organic solvents by laser flash photolysis (LFP) of the corresponding porphyrin-manganese(III) perchlorate and chlorate complexes, respectively, permitting direct kinetic studies. The porphyrin systems studied were 5,10,15,20-tetraphenylporphyrin (TPP), 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin (TPFPP), and 5,10,15,20-tetrakis(4-methylpyridinium)porphyrin (TMPyP). The order of reactivity for (porphyrin)Mn(V)(O) derivatives in self-decay reactions in acetonitrile and in oxidations of substrates was (TPFPP) > (TMPyP) > (TPP). Representative rate constants for reaction of (TPFPP)Mn(V)(O) in acetonitrile are k = 6.1 x 10(5) M(-1) s(-1) for cis-stilbene and k = 1.4 x 10(5) M(-1) s(-1) for diphenylmethane, and the kinetic isotope effect in oxidation of ethylbenzene and ethylbenzene-d(10) is k(H)/k(D) = 2.3. Competitive oxidation reactions conducted under catalytic conditions display approximately the same relative rate constants as were found in the LFP studies of (porphyrin)Mn(V)(O) derivatives. The apparent rate constants for reactions of (porphyrin)Mn(IV)(O) species show inverted reactivity order with (TPFPP) < (TMPyP) < (TPP) in reactions with cis-stilbene, triphenylamine, and triphenylphosphine. The inverted reactivity results because (porphyrin)Mn(IV)(O) disproportionates to (porphyrin)Mn(III)X and (porphyrin)Mn(V)(O), which is the primary oxidant, and the equilibrium constants for disproportionation of (porphyrin)Mn(IV)(O) are in the order (TPFPP) < (TMPyP) < (TPP). The fast comproportionation reaction of (TPFPP)Mn(V)(O) with (TPFPP)Mn(III)Cl to give (TPFPP)Mn(IV)(O) (k = 5 x 10(8) M(-1) s(-1)) and disproportionation reaction of (TPP)Mn(IV)(O) to give (TPP)Mn(V)(O) and (TPP)Mn(III)X (k approximately 2.5 x 10(9) M(-1) s(-1)) were observed. The relative populations of (porphyrin)Mn(V)(O) and (porphyrin)Mn(IV)(O) were determined from the ratios of observed rate constants for self-decay reactions in acetonitrile and oxidation reactions of cis-stilbene by the two oxo derivatives, and apparent disproportionation equilibrium constants for the three systems in acetonitrile were estimated. A model for oxidations under catalytic conditions is presented.


Sustained oscillations in extracellular calcium concentrations upon hormonal stimulation of perfused rat liver.
Sustained oscillations in extracellular free Ca2+ were shown to occur on addition of vasopressin, phenylephrine or angiotensin II in isolated rat liver perfused with low (10 microM)-Ca2+ medium. The amplitude and frequency of oscillation depend on hormone concentration. In contrast, Ca2+ releases on addition of ATP, t-butyl hydroperoxide or arachidonate do not exhibit oscillatory behaviour. The vasopressin-induced oscillations were suppressed by glucagon and dibutyryl 3',5'-cyclic AMP, but not by dibutyryl 3',5'-cyclic GMP. These observations in the extracellular space complement observations by Woods, Cuthbertson & Cobbold [(1986) Nature (London) 319, 600-602] on oscillations in intracellular free Ca2+ in single liver cells.


Trauma-Associated Pulmonary Laceration in Dogs-A Cross Sectional Study of 364 Dogs.
In this study, we describe the computed tomography (CT) features of pulmonary laceration in a study population, which included 364 client-owned dogs that underwent CT examination for thoracic trauma, and compared the characteristics and outcomes of dogs with and without CT evidence of pulmonary laceration. Lung laceration occurred in 46/364 dogs with thoracic trauma (prevalence 12.6%). Dogs with lung laceration were significantly younger than dogs in the control group (median 42 months (interquartile range (IQR) 52.3) and 62 months (IQR 86.1), respectively; p = 0.02). Dogs with lung laceration were significantly heavier than dogs without laceration (median 20.8 kg (IQR 23.3) and median 8.7 kg (IQR 12.4 kg), respectively p < 0.0001). When comparing groups of dogs with thoracic trauma with and without lung laceration, the frequency of high-energy motor vehicle accident trauma was more elevated in dogs with lung laceration than in the control group. No significant differences were observed between groups regarding tge frequency and length of hospitalization and 30-day mortality. Similar to the human classification scheme, four CT patterns are described in dogs in this study: Type 1, large pulmonary laceration located deeply in the pulmonary parenchyma or around an interlobar fissure; Type 2, laceration occurring in the paraspinal lung parenchyma, not associated with vertebral fracture; Type 3, subpleural lung laceration intimately associated with an adjacent rib or vertebral fracture; Type 4, subpleural lesions not associated with rib fractures. Complications were seen in 2/46 dogs and included lung abscess and collapse.


Habitat-performance relationships: finding the right metric at a given spatial scale.
The field of habitat ecology has been muddled by imprecise terminology regarding what constitutes habitat, and how importance is measured through use, selection, avoidance and other bio-statistical terminology. Added to the confusion is the idea that habitat is scale-specific. Despite these conceptual difficulties, ecologists have made advances in understanding 'how habitats are important to animals', and data from animal-borne global positioning system (GPS) units have the potential to help this clarification. Here, we propose a new conceptual framework to connect habitats with measures of animal performance itself--towards assessing habitat-performance relationship (HPR). Long-term studies will be needed to estimate consequences of habitat selection for animal performance. GPS data from wildlife can provide new approaches for studying useful correlates of performance that we review. Recent examples include merging traditional resource selection studies with information about resources used at different critical life-history events (e.g. nesting, calving, migration), uncovering habitats that facilitate movement or foraging and, ultimately, comparing resources used through different life-history strategies with those resulting in death. By integrating data from GPS receivers with other animal-borne technologies and combining those data with additional life-history information, we believe understanding the drivers of HPRs will inform animal ecology and improve conservation.