[Prevalence of dyslipidemias in adults in laboratory tests from Salvador, Brazil].
To determine the prevalence of dyslipidemia in non-hospital laboratory tests of adults from Salvador, Brazil. The study was carried out in subjects from a probabilistic sample of 25% of a total of 104 local laboratories that used the same enzymatic method for lipid analysis with the quality control as recommended by the Brazilian Society of Clinical Analyses. These represented 93% of all non-hospital laboratories of Salvador in 1995. The odd months of 1995 were selected for sampling in the present study. Criteria for dyslipidemias were: total cholesterol > or = 240; LDL > or = 160; HDL < 35 and triglycerides > or = 200 mg/dl. Prevalence rates and their 95% confidence intervals (CI) and chi 2 test were used in the analyses. Tests from two of the 26 laboratories were not used in the analyses due to file problems or refusal. 7,392 adults were screened, 65% female. We observed prevalence (95% CI) of hypercholesterolemia in 30.0 (27.8; 32.2)%, high LDL in 30.1 (30.8; 35.4)% and hypertriglyceridemia in 30.4 (29.0; 31.4)% of the females. As to the male subjects, prevalences were; 24.0 (20.5; 27.5)% for hypercholesterolemia, 26.1 (22.4; 29.3)% for high LDL, 27.6 (25.7; 29.5)% for hypertriglyceridemia. All gender differences were significant. Low HDL occurred in 15.9 (14.2; 17.8)% of males and in 8.0 (7.1; 8.9)% of females. Dyslipidemia is an important risk factor observed in non-hospital laboratory tests of men and women in Salvador. Our data may provide physicians and other health care professionals with objective information to encourage life-style changes.


Role of l- and d-Menthol in the Glucuronidation and Detoxification of the Major Lung Carcinogen, NNAL.
Menthol, which creates mint flavor and scent, is often added to tobacco in both menthol and nonmenthol cigarettes. A potent tobacco carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), is extensively metabolized to its equally carcinogenic metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) as (R)- or (S)-NNAL enantiomers. NNAL is detoxified by UDP-glucuronosyltransferase (UGT) enzymes, with glucuronidation occurring on either NNAL's pyridine ring nitrogen (NNAL-N-Gluc) or the chiral alcohol [(R)- or (S)-NNAL-O-Gluc]. To characterize a potential effect by menthol on NNAL glucuronidation, in vitro menthol glucuronidation assays and menthol inhibition of NNAL-Gluc formation assays were performed. Additionally, NNAL and menthol glucuronides (MG) were measured in the urine of smokers (n = 100) from the Southern Community Cohort Study. UGTs 1A9, 1A10, 2A1, 2A2, 2A3, 2B4, 2B7, and 2B17 all exhibited glucuronidating activity against both l- and d-menthol. In human liver microsomes, both l- and d-menthol inhibited the formation of each NNAL-Gluc, with a stereospecific difference observed between the formation of (R)-NNAL-O-Gluc and (S)-NNAL-O-Gluc in the presence of d-menthol but not l-menthol. With the exception of three nonmenthol cigarette smokers, urinary MG was detected in all menthol and nonmenthol smokers, with l-MG comprising >98% of total urinary MG. Levels of urinary NNAL-N-Gluc were significantly (P < 0.05) lower among subjects with high levels of total urinary MG; no significant changes in free NNAL were observed. These data suggest that the presence of menthol could lead to increases in alternative, activating metabolic pathways of NNAL in tobacco target tissues, increasing the opportunity for NNAL to damage DNA and lead to the development of tobacco-related cancers. SIGNIFICANCE STATEMENT: High levels of the major menthol metabolite, menthol-glucuronide, was observed in the urine of smokers of either menthol or nonmenthol cigarettes. The fact that a significant inverse correlation was observed between the levels of urinary menthol-glucuronide and NNAL-N-glucuronide, a major detoxification metabolite of the tobacco carcinogen, NNK, suggests that menthol may inhibit clearance of this important tobacco carcinogen.


Effects of nasal saline lavage on pediatric sinusitis symptoms and disease-specific quality of life: a case series of 10 patients.
We conducted a prospective study to assess (1) the effects of daily nasal irrigation in children with chronic sinonasal symptoms and (2) the impact that treatment had on disease-specific quality of life as assessed by a validated instrument, the five-item Sinus and Nasal Quality of Life Survey (SN-5), and by an overall nasal quality-of-life (NQL) score based on a 10-point faces scale. Our patient population was made up of 10 children-7 girls and 3 boys, aged 3 to 9 years (mean: 6.1)-who had presented with symptoms of chronic rhinosinusitis for more than 3 months and who had not responded to previous medical management. Patients were administered nasal saline lavage daily for 1 month. These patients and/or their caregivers completed an SN-5 questionnaire upon entry into the study and at the completion of treatment. At study's end, a comparison of pre- and post-treatment scores with paired Student t tests showed that the mean total SN-5 score improved significantly over baseline, falling 45% from 21.4 to 11.7 (p = 0.0002). Moreover, significant overall improvement was seen in each of the five subcategories of the SN-5 survey (p = 0.0009 to 0.038). The NQL scores also improved significantly from 4.7 to 7.7 (p = 0.0034). Compliance with nasal lavage was generally good among the 10 patients, as 8 of them used at least 75% of the recommended quantity of saline at least once a day. During a follow-up period that ranged from 2 to 23 months (mean: 10.4), only 1 patient required an adenoidectomy for symptom control. The results of this pilot study suggest that nasal saline lavage may significantly alleviate chronic sinonasal symptoms and improve disease-specific quality of life in children with symptoms of chronic rhinosinusitis.


Fluorescence resonance energy transfer scanning near-field optical microscopy.
The method of fluorescence resonance energy transfer scanning near-field optical microscopy (FRET SNOM) consists in the separation of a FRET pair between an SNOM tip and a sample. The donor (or acceptor) centre is located at the tip apex and scanned in the vicinity of a sample while acceptor fluorescence (or donor-fluorescence quenching) is detected. It is shown that the spatial resolution for such an approach is governed not by the aperture size but by the FRET characteristic radius (Förster radius), and thus can attain the value of 2-7 nm with the same (or higher) sensitivity as characteristic for the aperture SNOM. The theoretical fundamentals of the method, its experimental realization and connections with other types of near-field optical microscopy are discussed. Coherent FRET SNOM, which can be realized at liquid helium temperatures, and its possible applications for quantum informatics, are briefly outlined.


IL-10 protects against OPC ferroptosis by regulating lipid reactive oxygen species levels post stroke.
Accumulation of reactive oxygen species (ROS), especially on lipids, induces massive cell death in neurons and oligodendrocyte progenitor cells (OPCs) and causes severe neurologic deficits post stroke. While small compounds, such as deferoxamine, lipostatin-1, and ferrostatin-1, have been shown to be effective in reducing lipid ROS, the mechanisms by which endogenously protective molecules act against lipid ROS accumulation and subsequent cell death are still unclear, especially in OPCs, which are critical for maintaining white matter integrity and improving long-term outcomes after stroke. Here, using mouse primary OPC cultures, we demonstrate that interleukin-10 (IL-10), a cytokine playing roles in reducing neuroinflammation and promoting hematoma clearance, significantly reduced hemorrhage-induced lipid ROS accumulation and subsequent ferroptosis in OPCs. Mechanistically, IL-10 activated the IL-10R/STAT3 signaling pathway and upregulated the DLK1/AMPK/ACC axis. Subsequently, IL-10 reprogrammed lipid metabolism and reduced lipid ROS accumulation. In addition, in an autologous blood injection intracerebral hemorrhagic stroke (ICH) mouse model, deficiency of the endogenous Il-10, specific knocking out Il10r or Dlk1 in OPCs, or administration of ACC inhibitor was associated with increased OPC cell death, demyelination, axonal sprouting, and the cognitive deficits during the chronic phase of ICH and vice versa. These data suggest that IL-10 protects against OPC loss and white matter injury by reducing lipid ROS, supporting further development of potential clinical applications to benefit patients with stroke and related disorders.


Genomic and proteomic profiles reveal the association of gelsolin to TP53 status and bladder cancer progression.
Bladder cancer transformation and immortalization require the inactivation of key regulatory genes, including TP53. Genotyping of a large cohort of bladder cancer patients (n = 256) using the TP53 GeneChip showed mutations in 103 cases (40.2%), the majority of them mapping to the DNA-binding core domain. TP53 mutation status was significantly associated with tumor stage (P = 0.0001) and overall survival for patients with advanced disease (P = 0.01). Transcript profiling using oligonucleotide arrays was performed on a subset of these cases (n = 46). Supervised analyses identified genes differentially expressed between invasive bladder tumors with wild-type (n = 24) and mutated TP53 (n = 22). Pathway analyses of top-ranked genes supported the central role of TP53 in the functional network of such gene patterns. A proteomic strategy using reverse phase arrays with protein extracts of bladder cancer cell lines validated the association of identified differentially expressed genes, such as gelsolin, to TP53 status. Immunohistochemistry on tissue microarrays (n = 294) revealed that gelsolin was associated with tumor stage and overall survival, correlating positively with TP53 status in a subset of these patients. This study further reveals that TP53 mutations are frequent events in bladder cancer progression and identified gelsolin related to TP53 status, tumor staging, and clinical outcome by independent high-throughput strategies.


A Molecular Framework for the Control of Adventitious Rooting by TIR1/AFB2-Aux/IAA-Dependent Auxin Signaling in Arabidopsis.
In Arabidopsis thaliana, canonical auxin-dependent gene regulation is mediated by 23 transcription factors from the AUXIN RESPONSE FACTOR (ARF) family that interact with auxin/indole acetic acid repressors (Aux/IAAs), which themselves form co-receptor complexes with one of six TRANSPORT INHIBITOR1/AUXIN-SIGNALLING F-BOX (TIR1/AFB) proteins. Different combinations of co-receptors drive specific sensing outputs, allowing auxin to control a myriad of processes. ARF6 and ARF8 are positive regulators of adventitious root initiation upstream of jasmonate, but the exact auxin co-receptor complexes controlling the transcriptional activity of these proteins has remained unknown. Here, using loss-of-function mutants we show that three Aux/IAA genes, IAA6, IAA9, and IAA17, act additively in the control of adventitious root (AR) initiation. These three IAA proteins interact with ARF6 and/or ARF8 and likely repress their activity in AR development. We show that TIR1 and AFB2 are positive regulators of AR formation and TIR1 plays a dual role in the control of jasmonic acid (JA) biosynthesis and conjugation, as several JA biosynthesis genes are up-regulated in the tir1-1 mutant. These results lead us to propose that in the presence of auxin, TIR1 and AFB2 form specific sensing complexes with IAA6, IAA9, and/or IAA17 to modulate JA homeostasis and control AR initiation.


Modelling associations between public understanding, engagement and forest conditions in the Inland Northwest, USA.
Opinions about public lands and the actions of private non-industrial forest owners in the western United States play important roles in forested landscape management as both public and private forests face increasing risks from large wildfires, pests and disease. This work presents the responses from two surveys, a random-sample telephone survey of more than 1500 residents and a mail survey targeting owners of parcels with 10 or more acres of forest. These surveys were conducted in three counties (Wallowa, Union, and Baker) in northeast Oregon, USA. We analyze these survey data using structural equation models in order to assess how individual characteristics and understanding of forest management issues affect perceptions about forest conditions and risks associated with declining forest health on public lands. We test whether forest understanding is informed by background, beliefs, and experiences, and whether as an intervening variable it is associated with views about forest conditions on publicly managed forests. Individual background characteristics such as age, gender and county of residence have significant direct or indirect effects on our measurement of understanding. Controlling for background factors, we found that forest owners with higher self-assessed understanding, and more education about forest management, tend to hold more pessimistic views about forest conditions. Based on our results we argue that self-assessed understanding, interest in learning, and willingness to engage in extension activities together have leverage to affect perceptions about the risks posed by declining forest conditions on public lands, influence land owner actions, and affect support for public policies. These results also have broader implications for management of forested landscapes on public and private lands amidst changing demographics in rural communities across the Inland Northwest where migration may significantly alter the composition of forest owner goals, understanding, and support for various management actions.


Production of hepatitis B virus covalently closed circular DNA in transfected cells is independent of surface antigen synthesis.
Covalently closed circular DNA (cccDNA) is the first hepatitis B virus (HBV) DNA replicative intermediate formed from the genomic DNA and serves as the template for synthesis of viral mRNA and pregenomic RNA. It also appears to be produced by intracellular recycling of relaxed circular DNA intermediates. Here, we report that none of the forms of HBV surface antigen affect this intracellular recycling of HBV DNA. Elimination of the initiation codons for the large and middle surface proteins did not affect the detection of surface antigen in culture supernatants. In contrast, detection of surface antigen was eliminated by the removal of, or the introduction of two stop codons downstream of, the initiation codon of the small (major) surface antigen. None of the mutations affected the production, in transfected HepG2 cells, of HBV DNA replicative intermediates, including cccDNA.


Complications and outcomes associated with 13 cases of triceps tendon disruption in dogs and cats (2003-2014).
This study reports data from a larger number of cases of triceps tendon disruption. Records from 10 veterinary referral hospitals between 2003 and 2014 were searched for canine and feline cases diagnosed with triceps tendon disruption, based on orthopaedic examination confirmed during surgery. Long-term follow-up and owner satisfaction were assessed using a questionnaire. There were 13 cases of triceps tendon disruption diagnosed across seven hospitals (nine dogs, four cats). Trauma, history or presence of a wound, surgery in the region of tendon attachment or corticosteroid treatment preceded triceps tendon disruption. Radiographic signs or histopathology suggestive of a chronic tendinopathy was common. All cases underwent surgical repair involving a tendon suture pattern, 12 of which were secured through bone tunnels. Immobilisation was used in all cases in the form of transarticular external skeletal fixation (TAESF) (8/9 dogs) or spica splint (four cats, two dogs; in one dog a TAESF was applied after complications associated with the spica splint). Complications occurred in 11 cases (17 total complications), frequently associated with the immobilisation method. One case had traumatic tendon rerupture two years following surgery. A wound at presentation was associated with the development of multiple complications. Nine cases had long-term follow-up; five achieved normal function, four achieved acceptable function. Despite the complications, overall return to subjective normal or acceptable function, as assessed by the owners, was achieved in the majority of cases.


Palivizumab for prophylaxis against respiratory syncytial virus infection in children with cystic fibrosis.
Respiratory syncytial virus (RSV) infection causes acute lung infection in infants and young children worldwide, resulting in considerable morbidity and mortality. Children with cystic fibrosis (CF) are prone to recurrent lung inflammation, bacterial colonisation and subsequent chronic airway disease, putting them at risk for severe RSV infections requiring intensive care and respiratory support. No treatment currently exists, hence prevention is important. Palivizumab is effective in reducing RSV hospitalisation rates and is recommended for prophylaxis in high-risk children with other conditions. It is unclear if palivizumab can prevent RSV hospitalisations and intensive care unit admissions in children with CF. To determine the efficacy and safety of palivizumab (Synagis((R))) compared with placebo, no prophylaxis or other prophylaxis, in preventing hospitalisation and mortality from RSV infection in children with CF. We searched the Cochrane CF and Genetic Disorders Group Trials Register and scanned references of the eligible study and related reviews.Last search: 20 January 2010. Randomised and quasi-randomised studies. The authors independently extracted data and assessed risk of bias. One study (186 infants up to two years old) comparing five monthly doses of palivizumab (N=92) to placebo (N=94) over one RSV season was identified and met our inclusion criteria. At six months follow-up, one participant in each group was hospitalised due to RSV; there were no deaths in either group. In the palivizumab and placebo groups, 86 and 90 children experienced any adverse event, while 5 and 4 children had related adverse events respectively. Nineteeen children receiving palivizumab and 16 receiving placebo suffered serious adverse events; one participant receiving palivizumab discontinued due to this. At 12 months follow-up, there were no significant differences between groups in number of Pseudomonas bacterial colonisations or change in weight-to-height ratio. We identified one randomised controlled trial comparing five monthly doses of palivizumab to placebo in infants up to two years old with CF. While the overall incidence of adverse events was similar in both groups, it is not possible to draw conclusions on the safety and tolerability of RSV prophylaxis with palivizumab in infants with CF because the trial did not specify how adverse events were classified. Six months after treatment, the authors reported no clinically meaningful differences in outcomes; however no data were provided. Additional randomised studies are needed to establish the safety and efficacy of palivizumab in children with CF.


Hematopoietic cell transplantation in acute lymphoblastic leukemia: better long term event-free survival with conditioning regimens containing total body irradiation.
There is only limited experience with conditioning regimens based on busulfan for patients with acute lymphoblastic leukemia (ALL). Therefore, the aim of this study was to compare the event-free survival (EFS), transplant-related mortality (TRM) and the probability of relapse (PR) of patients undergoing hematopoietic cell transplantation (HCT) for ALL conditioned with or without total body irradiation (TBI). The study sample consisted of 156 patients conditioned with regimens based on TBI (n=114) or on high doses of oral busulfan (BU) (n=42). Most of the BU group received phenytoin as prophylaxis for seizures. The median follow-up was 6 years. EFS at 6 years was 43% (95% CI 35%-51%) versus 22% (95% CI 10%-34%) in the TBI and BU subsets respectively (p=0.01). TRM at 18 months was 22% and 17% in the BU and TBI groups (p=0.24), respectively. At 3 years actuarial PR was 71% in the BU group and 47% in the TBI group (p=0.01). In the multivariable analysis, a worse EFS was associated with BU, relative risk (RR) 1.7; advanced disease versus 1st and 2nd complete remission (CR) at HCT, RR 2.5; absence of chronic graft-versus-host disease, RR 1.8; development of veno-occlusive disease RR 2.2 and shorter CR duration before transplant. INTERPRETATION AND CONCLUSIONS. TBI was associated with a lower relapse rate and better EFS, even in patients in 1(st )and 2(nd) CR, than schemes based on high doses of busulfan. This suggests that conditioning regimens based on TBI should remain the standard method of preparative regimen for patients with ALL.


UV-visible-NIR reflectance spectrophotometry in cultural heritage: Background paper.
Diffuse reflectance spectrophotometry in the ultraviolet, visible and near-infrared (UV-vis-NIR) range is becoming a widely-used tool for the non-invasive analysis of cultural heritage objects, especially in its variant with optical fibres, commonly dubbed FORS (Fibre Optic Reflectance Spectroscopy). It is especially useful to study objects such as medieval manuscripts for which sampling is prohibited by current conservation standards. This brief describes the most common types of application of FORS in the analysis of cultural heritage objects; it also illustrates the instrumentation used and the main advantages and limitations of the technique.


Gastroenteropancreatic endocrine tumours: effect of Sandostatin on tumour growth. The German Sandostatin Study Group.
One hundred and fifteen gastroenteropancreatic (GEP) patients with malignant endocrine tumours entered a prospective multicentre trial (12 patients with gastrinoma, 53 with carcinoid syndrome, 45 with nonfunctioning tumours and 5 with other endocrine GEP tumours) to determine the efficacy of 200 micrograms Sandostatin t.i.d. in the control of tumour growth. This interim report describes the results in 85 patients. Thirty-four patients died, 14 before and 20 after the first follow-up investigation, indicating a 'negative' selection of patients included in the trial and suggesting that Sandostatin is unable to prevent disease progression when it is far advanced. In the evaluation of 68 patients followed up for at least 3 months, partial regression was observed in 4.4%, stable disease in 50% and tumour progression in 45%. An initially favourable response occurred frequently, however, it was followed by a decrease in response, from 54.4% at 3 months to 38% at 12 months, for the whole group of patients. Proven inhibition of tumour growth was mirrored by suppression of serum and urine hormone parameters. It is concluded that Sandostatin exerts a beneficial effect on tumour growth in patients with metastatic endocrine GEP tumours. This beneficial effect decreases with time and is as yet unpredictable in the individual patient.


Duration of type 2 diabetes and very low density lipoprotein levels are associated with cognitive dysfunction in metabolic syndrome.
Type 2 diabetes (T2D) is now recognized as an independent risk factor for accelerated cognitive decline and neurological conditions like Alzheimer's disease. Less is known about the neurocognitive function of T2D patients with comorbid metabolic syndrome, despite their elevated risk for impairment. Computerized testing in 47 adults with T2D that met criteria for NCEP metabolic syndrome revealed that cognitive impairment was prevalent, including 13% in tests of memory, 50% in attention, and 35% in executive function. Partial correlations showed that longer duration of diabetes was associated with poorer performance on tests of basic attention (r = -0.43), working memory (r = 0.43), and executive function (r = 0.42). Strong associations between very low density lipoprotein and poor cognitive function also emerged, including tests of set shifting (r = 0.47) and cognitive inhibition (r = -0.51). Findings suggest that patients with T2D that meet criteria for metabolic syndrome are at high risk for cognitive impairment. Prospective studies should look to replicate these findings and examine the possible neuroprotective effects of lipid-lowering medication in this population.


Rehabilitation therapies after botulinum toxin-A injection to manage limb spasticity: a systematic review.
Botulinum toxin A (BoNT-A) injections are increasingly used to treat muscle spasticity and are often complemented by adjunctive rehabilitation therapies; however, little is known about the effect of therapy after injection. The aim of this study was to identify and summarize evidence on rehabilitation therapies used after BoNT-A injections to improve motor function in adults with neurological impairments. Searches were conducted in PubMed, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), CINAHL, National Research Register, metaRegistry of Controlled Trials, PEDro, and OTseeker. Randomized and quasi-randomized controlled trials were considered for inclusion. Participants with neurological impairments received BoNT-A to treat focal spasticity in limbs, with rehabilitation interventions provided to experimental groups only. Primary outcome measures were joint mobility, function of the affected limb, and spasticity. Eleven studies with 234 participants, most of whom had stroke, were included in the review. Two reviewers extracted study details and data. Methodological quality was rated using the PEDro scale. Both fixed-effects and random-effects models were used to calculate effect size. Studies were of variable quality: 3 were poor (PEDro score 1 to 4), and 8 were moderate (PEDro score 6 to 7). No study investigated effects for longer than 24 weeks (6 months). Included trials presented 9 therapy types, including ergometer cycling, electrical stimulation, stretch (casting, splinting, taping, or manual or exercise-induced stretch), constraint-induced movement therapy, task-specific motor training, and exercise programs. Statistical findings suggest that combined therapy and BoNT-A is slightly more effective than BoNT-A alone. Evidence relating to impact of adjunct therapy is available, but the heterogeneity of studies limits the opportunity to demonstrate overall impact. Researchers need to consider the benefits of greater consistency in study approaches and measures so that meaningful evaluations of overall adjunct therapy effects can be made.


The cost-effectiveness of large amalgam and crown restorations over a 10-year period.
To assist clinical decision making for an individual patient or on a community level, this study was done to determine the differences in costs and effectiveness of large amalgams and crowns over 5 and 10 years when catastrophic subsequent treatment (root canal therapy or extraction) was the outcome. Administrative data for patients seen at the University of Iowa, College of Dentistry for 1735 large amalgam and crown restorations in 1987 or 1988 were used. Annual costs and effectiveness values were calculated. Costs of initial treatment (large amalgam or crown), and future treatments were determined, averaged and discounted. The effectiveness measure was defined as the number of years a tooth remained in a state free of catastrophic subsequent treatment. Years free of catastrophic treatment were averaged, and discounted. The years free of catastrophic treatment accounted for individuals who dropped out or withdrew from the study. Teeth with crowns had higher effectiveness values at a much higher cost than teeth restored with large amalgams. The cost of an addition year free of catastrophic treatment for crowns was 1088.41 dollars at 5 years and 500.10 dollars at 10 years. Teeth in women had more favorable cost-effectiveness ratios than those in men, and teeth in the maxillary arch had more favorable cost-effectiveness ratios than teeth in the mandibular arch. Neither the large amalgam or crown restoration had both the lowest cost and the highest effectiveness. The higher incremental cost-effectiveness ratio for crowns should be considered when making treatment decisions between large amalgam and crown restorations.


Antioxidant capacity of crude extracts containing carotenoids from the berries of various cultivars of Sea buckthorn (Hippophae rhamnoides L.).
Comparative analysis of antioxidant capacity was performed using FRAP and DPPH methods on extracts containing carotenoids acquired from fruits of Sea buckthorn. The examination included nine varieties of Sea buckthorn growing in the comparative cultivation. Conducted analysis allowed to compare the antioxidant capacity with carotenoids content measured with spectrophotometric and HPLC methods. Three of the examined cultivars indicating high antioxidant activity in both, FRAP and DPPH methods, also revealed highest ('Aromatnaya') and high ('Botanicheskaya', 'Arumnyj') total carotenoids content in HPLC analysis.


HIV update. Diagnostic tests and markers of disease progression and response to therapy.
HIV-1 infection continues to present diagnostic and management challenges on a global scale. Multiple testing strategies currently are used for evaluation of patients and infection status. Serologic testing represents the standard for diagnosis of HIV-1 infection. Molecular methods enable screening of the blood supply, detection of vertical transmission, and viral load testing to monitor patients during therapy. Molecular resistance testing facilitates the assessment of viral quasi species and the presence of specific mutations that confer antiviral resistance. Both genotyping and phenotyping approaches have been adopted for HIV-1 drug-resistance testing and enable rational modification of therapies for patients in whom their current regimens are failing.


A computational structural analysis of functional attributes of hypodermin A and B proteins: A way forward for vaccine development.
Hypodermosis is a parasitic disease of cattle. The pathogenicity of the disease is attributed to Hypodermin proteins (Hypodermin A, Hypodermin B and Hypodermin C). Studies suggest that Hypodermin proteins may be defined as Serine proteases and collagenases. The structure of both proteases Hypodermin A and Hypodermin B were modeled using the Swiss-model server followed by its validation using Procheck, Errat and Verify-3D. Afterwards, both Hypodermin A and Hypodermin B were docked against collagen in order to study its interaction with respective Hypodermin proteins. The structure of both Hypodermin A and Hypodermin B showed more bent towards hydrophobic nature as more beta sheets were present in them. Both structures were also superimposed to check out similarities and differences present between them. Serine, Aspartic acid, Histidine, Glutamic acid and Lysine are found as interacting residues that are involved in hydrogen bonding with collagen. The interactions are found in the active domain region of Hypodermin proteins. The interacting residues were present in the active region of the hypodermin proteins thus validating the docking studies. This study may help in the drug development against hypodermosis with least side effects.


Dieldrin uptake and translocation in plants growing in hydroponic medium.
It has been known that the Cucurbitaceae family takes up a large amount of persistent organic pollutants from soils and that the translocation of those compounds in cucurbits is higher than those in non-cucurbits. To understand the persistent organic pollutant uptake mechanisms of plant species, we compared the dieldrin absorption and transportation potentials of several plants in hydroponic medium. Sorghum (Sorghum vulgare Moench), sunflower (Helianthus annuus L.), soybean (Glycine max), komatsuna (Brassica rapa var. peruviridis), white-flowered gourd (Lagenaria siceraria var. hispida), cucumber (Cucumis sativus L.), and zucchini (Cucurbita pepo L.) were grown in a dieldrin-added hydroponic medium for 10 d, and then the amount of dieldrin in their shoots and roots was measured. All of the roots contained dieldrin, whereas only the cucurbits (white-flowered gourd, cucumber, and zucchini) contained considerable amounts of dieldrin in their shoots. The dieldrin uptake to the roots depended on the concentration of the n-hexane soluble components in the roots, regardless of whether the dieldrin in the roots was translocated to shoots or not. The dieldrin uptake from the solution to the roots was thought to be due to a passive response, such as adsorption on the roots. The translocation of dieldrin from the roots to the shoots was probably through the xylems. The amounts of dieldrin in the shoots per transpiration rates were higher for cucurbits than for non-cucurbits. It seems likely that cucurbits have uptake mechanisms for hydrophobic organic chemicals.


Face Validity and Content Validity of a Game for Distal Radius Fracture Rehabilitation.
Background Patients recovering from a variety of wrist injuries are frequently advised to exercise to regain lost wrist and hand function. Treatment regimens to regain motion in the wrist are highly variable, and adherence to exercise protocols is known to be low. A serious game ReValidate! incorporating standardized exercise regimens was developed to motivate patients. In this study, the game is evaluated regarding its face validity and content validity. Methods In this cross-sectional study, a mixed group of "users" ( n = 53) including patients currently recovering from wrist injury, and a mixed group of "experts" ( n = 46) including professionals advising patients on therapy regimen after wrist injury played at least one complete level of the serious game. Players evaluated the game by means of a structured questionnaire regarding its content, clinical applicability, and user experience. Questions were answered on a Likert scale ranging from 1 (strongly disagree) to 5 (strongly agree). Results All groups valued the game as being able to support wrist rehabilitation and being of use to patients recovering from a distal radius fracture (users: median 4, P25-P75 3-4 vs. experts: median 4, P25-P75 3.50-5; p = not significant). The types of exercises performed during the game were considered to be both realistic and complete compared with regular physiotherapy exercises (users: median 4, P25-P75 3-4 vs. experts: median 4, P25-P75 3-5, p = not significant). Conclusions The ReValidate! serious game can be regarded as a valid tool for patients to regain their wrist function after injury. Level of evidence This is a Level II study.


Characterization of Ginkgo biloba Leaf Flavonoids as Neuroexocytosis Regulators.
Ginkgo biloba leaf (GBL) is known as a potential source of bioactive flavonoids, such as quercetin, arresting the neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-zippering. Here, the GBL flavonoids were isolated in two different manners and then examined for their bioactivity, physicochemical stability, and biocompatibility. The majority of flavonoids in the non-hydrolyzed and acidolyzed isolates, termed non-hydrolyzed isolate (NI) and acidolyzed isolate (AI) hereafter, were rich in flavonol glycosides and aglycones, respectively. Glycosidic/aglyconic quercetin and kaempferol were abundant in both NI and AI, whereas a little of apigenin, luteolin, and isorhamnetin were found in AI. NI was more thermostable in all pH ranges than quercetin, kaempferol, and AI. NI and AI both inhibited neurotransmitter release from differentiated neuronal PC-12 cells. NI and AI showed 1/2-1/3 lower EC50/CC50 values than quercetin and kaempferol. The NI and AI exhibited no toxicity assessed by the tests on chorioallantoic membranes of hen's eggs, removing toxicological concerns of irritation potential. Moreover, GBL isolates, particularly AI, showed antioxidant and anti-inflammatory activities in the use below the CC50 levels. Taken together, these results suggest that GBL isolates that are rich in antioxidant flavonoids are effective anti-neuroexocytotic agents with high stability and low toxicity.


Numerical analysis of aeroacoustic-structural interaction of a flexible panel in uniform duct flow.
Accurate prediction of the acoustics of fluid-structure interaction is important in devising quieting designs for engineering systems equipped with extensive flow duct networks where the thin duct wall panels are in contact with the flowing fluid. The flow unsteadiness generates acoustic waves that propagate back to the source region to modify the flow process generating them. Meanwhile the unsteady flow pressure excites the thin panels to vibrate, which in turn modifies the flow processes. Evidently a strong coupling between the fluid aeroacoustics and the panel structural dynamics exists. Such coupled physical processes have to be thoroughly understood; otherwise, effective quieting design is never achieved. This paper reports an analysis, using a time-domain numerical methodology the authors have recently developed, of the nonlinear aeroacoustic-structural interaction experienced by a flexible panel in a duct carrying a uniform mean flow. With no mean flow, the numerical results agree well with existing theories and reveal the physics of duct transmission loss. Four regimes of aeroacoustic-structural interaction are identified when the duct flow velocity increases from low subsonic to low supersonic values. Insight in the underlying physics of duct transmission loss at different velocities are highlighted and discussed.


Aberrant Clonal Hematopoiesis following Lentiviral Vector Transduction of HSPCs in a Rhesus Macaque.
Lentiviral vectors (LVs) are used for delivery of genes into hematopoietic stem and progenitor cells (HSPCs) in clinical trials worldwide. LVs, in contrast to retroviral vectors, are not associated with insertion site-associated malignant clonal expansions and, thus, are considered safer. Here, however, we present a case of markedly abnormal dysplastic clonal hematopoiesis affecting the erythroid, myeloid, and megakaryocytic lineages in a rhesus macaque transplanted with HSPCs that were transduced with a LV containing a strong retroviral murine stem cell virus (MSCV) constitutive promoter-enhancer in the LTR. Nine insertions were mapped in the abnormal clone, resulting in overexpression and aberrant splicing of several genes of interest, including the cytokine stem cell factor and the transcription factor PLAG1. This case represents the first clear link between lentiviral insertion-induced clonal expansion and a clinically abnormal transformed phenotype following transduction of normal primate or human HSPCs, which is concerning, and suggests that strong constitutive promoters should not be included in LVs.


Complications of elbow arthroscopy.
Although the potential complications of elbow arthroscopy, including nerve injuries, have been described, the prevalence of their occurrence has not been well defined. The purpose of this paper is to describe the serious and minor complications in a large series of patients treated with elbow arthroscopy. A retrospective review of 473 consecutive elbow arthroscopies performed in 449 patients over an eighteen-year period was conducted. Of the 473 cases, 414 were followed for more than six weeks. The most common final diagnoses were osteoarthritis (150 cases), loose bodies (112), and rheumatoid or inflammatory arthritis (seventy-five). The arthroscopic procedures included synovectomy (184), debridement of joint surfaces or adhesions (180), excision of osteophytes (164), diagnostic arthroscopy (154), loose-body removal (144), and capsular procedures such as capsular release, capsulotomy, and capsulectomy (seventy-three). A serious complication (a joint space infection) occurred after four (0.8%) of the arthroscopic procedures. Minor complications occurred after fifty (11%) of the arthroscopic procedures. These complications included prolonged drainage from or superficial infection at a portal site after thirty-three procedures, persistent minor contracture of 20 degrees or less after seven, and twelve transient nerve palsies (five ulnar palsies, four superficial radial palsies, one posterior interosseous palsy, one medial antebrachial cutaneous palsy, and one anterior interosseous palsy) in ten patients. The most significant risk factors for the development of a temporary nerve palsy were an underlying diagnosis of rheumatoid arthritis (p < 0.001) and a contracture (p < 0.05). There were no permanent neurovascular injuries, hematomas, or compartment syndromes in our series, and all of the minor complications, except for the minor contractures, resolved without sequelae. Our results indicate that the prevalence of temporary or minor complications following elbow arthroscopy may be greater than previously reported. However, serious or permanent complications were uncommon.


Effectiveness of saltwater baths in the treatment of epidermolysis bullosa.
Current management of epidermolysis bullosa (EB) focuses on preventing secondary infections--a leading cause of mortality--by regularly cleaning and bandaging blisters and erosions and preventive bandaging of high-friction areas. Unfortunately the baths and showers used to remove bandages and clean the skin are often painful, causing lack of adherence and subsequent increases in infections, pain, and antimicrobial use. This study evaluates the hypothesis that bathing individuals with EB in saltwater is less painful than in normal bath water. The study also explores whether taking saltwater baths reduces infections as measured through skin pruritus, odor, discharge, and nonbathing pain. Participants recruited from the Children's Hospital Colorado Outpatient EB Clinic completed standardized questionnaires assessing the effects of the clinic's saltwater bathing recommendations; the data were analyzed using frequencies and Fisher tests. After starting saltwater baths, patients reported a significant reduction in pain (91%), pain medication use (66%), skin odor (31%), and skin discharge (44%). No significant differences were found with respect to the type of EB, age, length of time using baths, or amount of salt added. Saltwater baths are a noninvasive, low-cost, effective treatment that significantly reduces bathing pain, pain medication use, and some signs of skin infection. This treatment can be recommended to patients with all studied EB types without regard to age, the specific amount of salt used, bathing frequency, or pain level. Given the central role bathing and dressing changes play in the management of EB, the use of saltwater baths can lead to significant improvement in quality of life.


Augmentation of Glucotoxicity, Oxidative Stress, Apoptosis and Mitochondrial Dysfunction in HepG2 Cells by Palmitic Acid.
Hyperglycemia and hyperlipidemia are the hallmarks of diabetes and obesity. Experimental and epidemiological studies have suggested that dietary management and caloric restriction are beneficial in reducing the complications of diabesity. Studies have suggested that increased availability of energy metabolites like glucose and saturated fatty acids induces metabolic, oxidative, and mitochondrial stress, accompanied by inflammation that may lead to chronic complications in diabetes. In the present study, we used human hepatoma HepG2 cells to investigate the effects of high glucose (25 mM) and high palmitic acid (up to 0.3 mM) on metabolic-, inflammatory-, and redox-stress-associated alterations in these cells. Our results showed increased lipid, protein, and DNA damage, leading to caspase-dependent apoptosis and mitochondrial dysfunction. Glucolipotoxicity increased ROS production and redox stress appeared to alter mitochondrial membrane potential and bioenergetics. Our results also demonstrate the enhanced ability of cytochrome P450s-dependent drug metabolism and antioxidant adaptation in HepG2 cells treated with palmitic acid, which was further augmented with high glucose. Altered NF-kB/AMPK/mTOR-dependent cell signaling and inflammatory (IL6/TNF-α) responses were also observed. Our results suggest that the presence of high-energy metabolites enhances apoptosis while suppressing autophagy by inducing inflammatory and oxidative stress responses that may be responsible for alterations in cell signaling and metabolism.


Brain barriers and their potential role in migraine pathophysiology.
Migraine is a ubiquitous neurologic disease that afflicts people of all ages. Its molecular pathogenesis involves peptides that promote intracranial vasodilation and modulate nociceptive transmission upon release from sensory afferents of cells in the trigeminal ganglion and parasympathetic efferents of cells in the sphenopalatine ganglion. Experimental data have confirmed that intravenous infusion of these vasoactive peptides induce migraine attacks in people with migraine, but it remains a point of scientific contention whether their site of action lies outside or within the central nervous system. In this context, it has been hypothesized that transient dysfunction of brain barriers before or during migraine attacks might facilitate the passage of migraine-inducing peptides into the central nervous system. Here, we review evidence suggestive of brain barrier dysfunction in migraine pathogenesis and conclude with lessons learned in order to provide directions for future research efforts.


Immunisation against ovine caseous lymphadenitis: comparison of Corynebacterium pseudotuberculosis vaccines with and without bacterial cells.
Sheep were immunised with Corynebacterium pseudotuberculosis vaccines prepared from cell-free toxoid or from toxoid with formalin-killed cells of C pseudotuberculosis added. Resistance of sheep to infection was tested 6 months after immunisation by inoculation with caseous lymphadenitis pus. The outcome was assessed 3 months later by slaughter and inspection of the sheep for lesions of caseous lymphadenitis. immunised sheep were adequately protected against infection as shown by a significant reduction in the number of sheep exhibiting lesions compared with control sheep, and by fewer abscesses in affected vaccinated sheep than in affected control sheep. The protective potency of the vaccines was not improved by the inclusion of cells of C pseudotuberculosis.


Nurses attitudes towards the importance of families in psychiatric care following an educational and training intervention program.
This study measures the attitudes of the psychiatric nurses, after having received an education and training intervention program (ETI-PROGRAM) in family systems nursing, towards the importance of the families in their care. Nurses' knowledge of the impact that family nursing intervention can have on family members may increase positive attitudes towards families. However, little is known about the impact that education and training intervention can have on nurses' attitudes, towards families in clinical practice. Quasi-experimental design was used to assess the change in nurses' attitudes towards families in psychiatric care after the intervention, which included a one-day seminar on the Calgary family nursing conceptual frameworks and skills training with clinical vignettes of families from psychiatry. The Families Importance in Nursing Care - Nurses' Attitude questionnaire was used to evaluate nurses' attitudes. A total of 81 nurses (65%) working in psychiatric care responded to the questionnaire. Nurses with more than 15 years of work experience were significantly more supportive of families in their care compared with less experienced nurses. Out of the 81 nurses, 52 (64%) answered the questionnaire again 14 months later. Furthermore, psychiatric nurses saw families significantly less burdensome after having participated in the ETI-PROGRAM.


Detection of long-range electrostatic interactions between charged molecules by means of fluorescence correlation spectroscopy.
In the present paper, an experimental feasibility study on the detection of long-range intermolecular interactions through three-dimensional molecular diffusion in solution is performed. This follows recent theoretical and numerical analyses reporting that long-range electrodynamic forces between biomolecules could be identified through deviations from Brownian diffusion. The suggested experimental technique was fluorescence correlation spectroscopy (FCS). By considering two oppositely charged molecular species in aqueous solution, namely, lysozymes and fluorescent dye molecules (Alexa488), the diffusion coefficient of the dyes has been measured for different values of the concentration of lysozyme, that is, for different average distances between the oppositely charged molecules. For our model, long-range interactions are of electrostatic origin, suggesting that their action radius can be varied by changing the ionic strength of the solution. The experimental outcomes clearly prove the detectability of long-range intermolecular interactions by means of the FCS technique. Molecular dynamics simulations provide a clear and unambiguous interpretation of the experimental results.


Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease in mice is influenced by the H-2D region: correlation with TEMV-specific delayed-type hypersensitivity.
Intracranial inoculation of Theiler's murine encephalomyelitis virus (TMEV) leads to the development of a chronic demyelinating disorder in certain mouse strains. Development of this disease is controlled by at least two unlinked genes, one of which is within or linked to the H-2 complex. In the present study, we attempted to map the relevant H-2 loci involved in susceptibility to TMEV-induced demyelination using crosses between SJL and several congenic H-2 recombinant mouse strains bearing different combinations of MHC genes from the susceptible H-2s and resistant H-2b haplotypes all on the C57BL/10 strain background. The data suggest that the D region of the H-2 complex strongly influences development of the demyelinating disease because increased susceptibility correlates well with homozygosity for H-2s alleles in the D region, but not in K or I-A. In addition, we also attempted to correlate certain immune and nonimmune pathophysiologic parameters with the development of clinical disease. Specifically, central nervous system TMEV titers and TMEV-specific humoral and cellular [delayed-type hypersensitivity (DTH) and T cell proliferative (Tprlf)] responses were examined. The data show that TMEV-induced demyelinating disease did not correlate with either CNS TMEV titers or TMEV-specific humoral or Tprlf responses but did correlate closely with the presence of high levels of TMEV-specific DTH. Collectively, our findings demonstrating a strong correlation between disease incidence, the presence of particular H-2D region genotypes, and high levels of TMEV-specific DTH in susceptible strains (as well as previous findings showing predominant mononuclear cell infiltrates in CNS demyelinating lesions) support the hypothesis that the disease is immune mediated rather than a result of direct cytolytic effects of virus infection.


Skin-to-Skin Contact Facilitates More Equal Parenthood - A Qualitative Study From Fathers' Perspective.
A descriptive design was used where 20 fathers of premature infants were interviewed using a semi-structured interview-guide. The guide contained the following five topics: the fathers' feelings about and experiences of using skin-to-skin contact (SSC), the physical environment at the Neonatal Intensive Care unit, staff attitudes, and whether SSC had any impact on the relationship with their partner and other children in the family. The interviews were analyzed using directed qualitative content analysis and the results compared with those of a qualitative systematic review by Anderzén-Carlsson and colleagues. The result shows that the fathers' overall experience of SSC was positive and as in the aforementioned review, they described experiences that were both gratifying and challenging. They felt more included and just as important as the mother because SSC and the demands placed on them by the premature birth helped them to achieve more equal parenthood. The fathers' overall experiences of SSC were positive as they felt included in their infants' care and just as important as the mothers. To a great degree the theoretical model tested was supported, although some weaknesses and differences were identified. It is important to include fathers in neonatal care and recognize that they feel equally important as mothers in caring for premature infants.


Written disclosure therapy for patients with chronic lung disease undergoing pulmonary rehabilitation.
Chronic lung diseases are typically associated with impaired quality of life, stress, and anxiety. Written disclosure therapy (WDT) reduces stress in patients with a variety of chronic illnesses. We sought to determine whether WDT benefits patients with chronic lung disease. A prospective, randomized, controlled trial was performed to evaluate the effect of using WDT in patients (N = 66) participating in a pulmonary rehabilitation program. Patients were randomly assigned to write about a particularly traumatic life event (WDT group) or to write about an emotionally neutral subject (control group). Exercise capacity, dyspnea and quality of life, and values of spirometry were recorded at baseline, at the end of the program, and at 6 months. The 6-minute walk distance (6MWD) significantly improved in both groups at 2 months, from 278 to 327 m in WDT and from 269 to 314 m in control groups (P < .01 in both groups). There was no difference in improvement in 6MWD between groups (P = .88). At 6 months, the gains made in 6MWD were no longer present. Dyspnea severity, as well as most of the other domains of the Chronic Respiratory Disease Questionnaire and the St. George's Respiratory Questionnaire, showed improvement within each group, but not between WDT and control groups. WDT did not add any additional benefit in patients with chronic obstructive pulmonary disease or idiopathic pulmonary fibrosis when included as a component of pulmonary rehabilitation. These results are in contrast to previously seen benefits in patients with asthma.


In Vitro Proliferation and Anti-Apoptosis of the Papain-Generated Casein and Soy Protein Hydrolysates towards Osteoblastic Cells (hFOB1.19).
Casein and soy protein were digested by papain to three degrees of hydrolysis (DH) 7.3%-13.3%, to obtain respective six casein and soy protein hydrolysates, aiming to clarify their in vitro proliferation and anti-apoptosis towards a human osteoblastic cell line (hFOB1.19 cells). Six casein and soy protein hydrolysates at five levels (0.01-0.2 mg/mL) mostly showed proliferation as positive 17β-estradiol did, because they conferred the osteoblasts with cell viability of 100%-114% and 104%-123%, respectively. The hydrolysates of higher DH values had stronger proliferation. Casein and soy protein hydrolysates of the highest DH values altered cell cycle progression, and enhanced cell proportion of S-phase from 50.5% to 56.5% and 60.5%. The two also antagonized etoposide- and NaF-induced osteoblast apoptosis. In apoptotic prevention, apoptotic cells were decreased from 31.6% to 22.6% and 15.6% (etoposide treatment), or from 19.5% to 17.7% and 12.4% (NaF treatment), respectively. In apoptotic reversal, soy protein hydrolysate decreased apoptotic cells from 13.3% to 11.7% (etoposide treatment), or from 14.5% to 11.0% (NaF treatment), but casein hydrolysate showed no reversal effect. It is concluded that the hydrolysates of two kinds had estradiol-like action on the osteoblasts, and soy protein hydrolysates had stronger proliferation and anti-apoptosis on the osteoblasts than casein hydrolysates.


Biochemical and Morphological Characterization of a Neurodevelopmental Disorder-Related Mono-ADP-Ribosylhydrolase, MACRO Domain Containing 2.
MACRO Domain Containing 2 (MacroD2) is a neurodevelopmental disorder-related mono-ADP-ribosylhydrolase. Molecular features of this protein in neural tissues are largely unknown. In this study, we generated a specific antibody against MacroD2, and carried out expression and morphological analyses of the molecule during mouse brain development. In Western blotting, 2 MacroD2 isoforms with molecular masses of ∼70 and ∼75 kDa started to be expressed at embryonic day 16.5, reached the maximal level at postnatal day 8, and then gradually decreased through P30. In contrast, other isoforms with molecular masses of ∼110 and ∼140 kDa gradually increased during embryonic to postnatal development. In immunohistochemical analyses, MacroD2 was strongly detected in cortical neurons in layer II-V at P0 and P7, while the protein expression decreased significantly in the neurons at P30. Immunofluorescence analyses revealed that MacroD2 was mainly distributed in the soma and to a lesser extent in the axon and dendrite of immature primary cultured mouse hippocampal neurons. On the other hand, in the matured hippocampal neurons, while MacroD2 was detected in the soma, it displayed in dendrites a punctate distribution pattern with a partial colocalization with synaptic markers, synaptophysin, and PSD95. The obtained results indicate that MacroD2 is expressed and may have a physiological role in the central nervous system during brain development.


Functional anatomy of the portal vein and its main and segmental branches in the adult man.
The AA. study the special disposition of collagen, elastic and muscular fibers in the supraduodenal portion of the portal vein and its main and segmental branches. Forty specimens were removed at necropsy from adult individuals and prepared by various methods: a) pellicle preparations stained by resorcin-fuchsin (Weigert), Azan, azo-carmine B, alum-carmine, or else examined by polarized light without staining, b) dissection under stereomicroscope of specimens treated by the method of Semper or van Gieson, c) histological sections stained by Masson's trichrome, Azan, Weigert's resorcin-fuchsin and Verhöff methods. The results corresponding to the different layers and levels of the vascular system studied are schematically reported in Figures 1A, 1C, 2A, 2E and 4B and are commented upon under a morphofunctional criterion.


[Validation study on a method for multiresidue analysis of pesticides in vegetables and fruits with supercritical fluid extraction].
Supercritical fluid extraction (SFE) was applied to extraction of pesticides from vegetables and fruits. Residues were extracted from homogenized samples mixed with water-absorbent polymer with supercritical carbon dioxide in a stainless steel tube, followed by elution with acetone. Co-extractives were removed by means of mini-column clean-up. Measurement was performed by GC-MS/MS. Calibration was achieved by preparing matrix-matched calibration standards to counteract matrix effects. With the Japanese method validation guideline as a reference, the method was assessed in 5 agricultural products spiked with 334 pesticides at 0.01 and 0.1 µg/g. Compounds at each level were extracted from 2 samples on 5 separate days. The trueness of the method for 189 pesticides in all samples was 70-120%, and the repeatability and within-run reproducibility were also consistent with the guideline. The trueness of the method for the other 71 pesticides was in the range of 50-70%, though the repeatability and within-run reproducibility were satisfactory. This method is available as a multiresidue analysis method for vegetables and fruits.


Diagnostic Dilemma in the Treatment of a Fatal Case of Bloody Diarrhea.
Although diarrhea is the most commonly reported pediatric illness in the United States, mortality is usually a rare and unexpected event. We report the case of a healthy 13-month-old male that succumbed to a diarrheal illness of unclear etiology. Presenting signs included frequent nonbloody stools that progressed to frankly bloody stools over 72 hours. Associated symptoms included fever, tenesmus, relief with stool passage, and significant fatigue. On examination, the patient appeared tired and lay with legs curled toward his chest. The abdominal exam was remarkable for hypoactive bowel sounds, diffuse tenderness to palpation without guarding or rebound pain, and intermittent prolapse of rectal tissue. Abdominal plain films demonstrated a paucity of bowel gas, especially in the rectum; and ultrasound revealed thickening of bowel loops in the left lower quadrant. Abdominal computed tomography scan showed decreased enhancement of the mucosa of the rectosigmoid colon. The patient deteriorated rapidly with cardiorespiratory arrest occurring 48 hours after admission. Despite a protracted effort at cardiopulmonary resuscitation, perfusing heart rate or rhythm could not be reestablished. Autopsy revealed infarction and necrosis of the rectosigmoid colon with invasive gram-negative bacilli. Here we present his perplexing case, diagnostic evaluations, and suggest a unifying diagnosis.


Fusobacterium nucleatum exacerbates the progression of pulpitis by regulating the STING-dependent pathway.
Bacterial infection is the main cause of pulpitis. However, whether a dominant bacteria can promote the progression of pulpitis and its underlying mechanism remains unclear. We provided a comprehensive assessment of the microbiota alteration in pulpitis using 16S rRNA sequencing. Fusobacterium nucleatum was the most enriched in pulpitis and played a pathogenic role accelerating pulpitis progression in rat pulpitis model. After odontoblast-like cells cocultured with F. nucleatum, the stimulator of interferon genes (STING) pathway and autophagy were activation. There was a float of STING expression during F. nucleatum stimulation. STING was degraded by autophagy at the early stage. At the late stage, F. nucleatum stimulated mitochondrial Reactive Oxygen Species (ROS) production, mitochondrial dysfunction and then mtDNA escape into cytosol. mtDNA, which escaped into cytosol, caused more cytosolic mtDNA binds to cyclic GMP-AMP synthase (cGAS). The release of IFN-β was dramatically reduced when mtDNA-cGAS-STING pathway inhibited. STING-/- mice showed milder periapical bone loss and lower serum IFN-β levels compared with wildtype mice after 28 days F. nucleatum-infected pulpitis model establishment. Our data demonstrated that F. nucleatum exacerbated the progression of pulpitis, which was mediated by the STING-dependent pathway.


Quantitative methods for analysing cumulative effects on fish migration success: a review.
It is often recognized, but seldom addressed, that a quantitative assessment of the cumulative effects, both additive and non-additive, of multiple stressors on fish survival would provide a more realistic representation of the factors that influence fish migration. This review presents a compilation of analytical methods applied to a well-studied fish migration, a more general review of quantitative multivariable methods, and a synthesis on how to apply new analytical techniques in fish migration studies. A compilation of adult migration papers from Fraser River sockeye salmon Oncorhynchus nerka revealed a limited number of multivariable methods being applied and the sub-optimal reliance on univariable methods for multivariable problems. The literature review of fisheries science, general biology and medicine identified a large number of alternative methods for dealing with cumulative effects, with a limited number of techniques being used in fish migration studies. An evaluation of the different methods revealed that certain classes of multivariable analyses will probably prove useful in future assessments of cumulative effects on fish migration. This overview and evaluation of quantitative methods gathered from the disparate fields should serve as a primer for anyone seeking to quantify cumulative effects on fish migration survival.


What is pain? A phenomenological approach to understanding.
This paper presents four phenomenological accounts of pain which describe: the pain at the time of the experience; the meaning the pain had at the time of the experience (reflection in); and the meaning of the experience looking back on it (reflection upon). Specifically, the accounts describe pain that is emotional, cognitive and physical (acute and chronic). By exploring the essence of pain using very personal accounts, we encourage nurses to reflect on how their own understanding of pain and individual responses to pain can impact on how they in turn recognise, assess and manage pain in older people and especially those living with dementia. Paper 1 highlights the under-assessment of pain in older people with dementia. By confronting readers with this paper's phenomenological approach, it is hoped that they will be challenged to address this situation.


[Health inequality among vulnerable groups in Mexico: older adults, indigenous people, and migrants].
Health vulnerability refers to a lack of protection for specific population groups with specific health problems, as well as the disadvantages they face in solving them in comparison with other population groups. This major public health problem has multiple and diverse causes, including a shortage of trained health care personnel and the lack of family, social, economic, and institutional support in obtaining care and minimizing health risks. Health vulnerability is a dynamic condition arising from the confluence of multiple social determinants. This article attempts to describe the health situation of three vulnerable groups in Mexico-older adults, indigenous people, and migrants-and, after defining the needs of each, explore measures that could contribute to the design and implementation of public health policies better tailored to their respective needs.


Haemodynamic effects of the antiarrhythmic quaternary ammonium compound QX-572 in man.
The haemodynamic effects of N, N-bis(phenyl-carbamoylmethyl) dimethylammonium chloride (QX-572) in man were studied. A controlled study was performed to rule out a possible influence of the catheterization procedure as such on the results. Ten patients with mild to moderate aortic regurgitation were studied: based on clinical data the patients were divided into 2 groups of 5. Randomly it was decided that one group should constitute a control group receiving saline while the second group received QX-572 , MG/KG BODY WEIGHT. In both groups the administration was performed as a slow intravenous infusion during 30 minutes. Heart rate, pressures in brachial artery and right atrium, cardiac output, stroke volume, and systemic vascular resistance were determined before, during, and up to 30 minutes after completion of placebo or QX-572. These variable remained stable in the control group while QX-572 produced an increase in heart rate most pronounced at the end of the infusion period. A transient decrease in systolic and mean brachial artery pressure during the infusion, and during the same period a decrease in right atrial pressure. Cardiac output and systemic vascular resistance were unchanged by QX-572 but they were not measured during the infusion when the changes in pressures were most pronounced. QX-572 was thought to act as a peripheral vasodilator during the infusion. Left ventricular contractility was studied by means of pressure curves obtained from a catheter tip manometer placed in the left ventricle. The first derivative of the isovolumic left ventricular pressure at the highest level (45mmHg) common to all patients was used (dp/dt-45). No significant difference could be observed when comparing mean changes of dp/dt-45 for the two groups. In the control group there was a slight but significant increase in dp/dt-45 during the time of observation. In the QX-572 group the results varied between individuals. Two of the patients differed from all other patients in the control and QX-572 groups showing a decrease in dp/dt-45 which, when most pronounced at the end of the infusion period, was -31 and -28 per cent of the preinfusion levels, respectively. This decrease probably reflects reduction of contractility. It was concluded that QX-572 in a dose of 8 mg/kg body weight did not have any major haemodynamic drawbacks.


The long and short flavodoxins: I. The role of the differentiating loop in apoflavodoxin structure and FMN binding.
Flavodoxins are well known one-domain alpha/beta electron-transfer proteins that, according to the presence or absence of a approximately 20-residue loop splitting the fifth beta-strand of the central beta-sheet, have been classified in two groups: long and short-chain flavodoxins, respectively. Although the flavodoxins have been extensively used as models to study electron transfer, ligand binding, protein stability and folding issues, the role of the loop has not been investigated. We have constructed two shortened versions of the long-chain Anabaena flavodoxin in which the split beta-strand has been spliced to remove the original loop. The two variants have been carefully analyzed using various spectroscopic and hydrodynamic criteria, and one of them is clearly well folded, indicating that the long loop is a peripheral element of the structure of long flavodoxins. However, the removal of the loop (which is not in contact with the cofactor in the native structure) markedly decreases the affinity of the apoflavodoxin-FMN complex. This seems related to the fact that, in long flavodoxins, the adjacent tyrosine-bearing FMN binding loop (which is longer and thus more flexible than in short flavodoxins) is stabilized in its competent conformation by interactions with the excised loop. The modest role played by the long loop of long flavodoxins in the structure of these proteins (and in its conformational stability, see Lopez-Llano, J., Maldonado, S., Jain, S., Lostao, A., Godoy-Ruiz, R., Sanchez-Ruiz, Cortijo, M., Fernandez-Recio, J., and Sancho, J. (2004) J. Biol. Chem. 279, 47184-47191) opens the possibility that its conservation in so many species is related to a functional role yet to be discovered. In this respect, we discuss the possibility that the long loop is involved in the recognition of some flavodoxin partners. In addition, we report on a structural feature of flavodoxins that could indicate that the short flavodoxins derive from the long ones.


Production and dosimetry of copper L ultrasoft x-rays for biological and biochemical investigations.
Ultrasoft x-rays provide a unique tool for investigating the intracellular mechanisms of radiation action. Secondary electrons are produced with a well defined energy and a range comparable with that of critical structures in the cell. Copper L characteristic x-rays of weighted average energy of 956 eV interact within the cell, mainly with the oxygen atom, typically producing a photoelectron with energy 424 eV (95%) followed by an Auger electron with an average energy of 505 eV, with a combined continuous slowing down approximation (csda) range of approximately 40 nm. The attenuation through the cell is similar to that of carbon K x-rays (277 eV, single electron), therefore a useful comparison can be made due to similar dose-averaging factors but different electron configurations (total range, and pairs versus singlets). The production, absorption, dosimetry and biological implications of Cu L x-rays using the Medical Research Council cold cathode source is described extending the number of energies available for study in the ultrasoft region. Design parameters were optimized to overcome the inherently low L-characteristic-to-bremsstrahlung yield ratio. Surface absorbed dose rates of 1 Gy min-1 have been obtained with a bremsstrahlung contamination of less than 0.5%. A confocal microscope was used to make thickness measurements on live cells to allow careful determination of the mean absorbed dose. Survival curves for V79-4 Chinese hamster cells were obtained, showing that Cu L x-rays are substantially more lethal per unit dose than are hard x-rays or gamma-rays, with a relative biological effectiveness (RBE) of 1.8. The data are consistent with the hypothesis that clustered damage at the DNA/chromatin level produced by low-energy electrons is biologically more effective.


Escherichia coli endonuclease VIII: cloning, sequencing, and overexpression of the nei structural gene and characterization of nei and nei nth mutants.
Escherichia coli possesses two DNA glycosylase/apurinic lyase activities with overlapping substrate specificities, endonuclease III and endonuclease VIII, that recognize and remove oxidized pyrimidines from DNA. Endonuclease III is encoded by the nth gene. Endonuclease VIII has now been purified to apparent homogeneity, and the gene, nei, has been cloned by using reverse genetics. The gene nei is located at 16 min on the E. coli chromosome and encodes a 263-amino-acid protein which shows significant homology in the N-terminal and C-terminal regions to five bacterial Fpg proteins. A nei partial deletion replacement mutant was constructed, and deletion of nei was confirmed by genomic PCR, activity analysis, and Western blot analysis. nth nei double mutants were hypersensitive to ionizing radiation and hydrogen peroxide but not as sensitive as mutants devoid of base excision repair (xth nfo). Single nth mutants exhibited wild-type sensitivity to X rays, while nei mutants were consistently slightly more sensitive than the wild type. Double mutants lacking both endonucleases III and VIII exhibited a strong spontaneous mutator phenotype (about 20-fold) as determined by a rifampin forward mutation assay. In contrast to nth mutants, which showed a weak mutator phenotype, nei single mutants behaved as the wild type.


Sugars that Glow in the Dark: Fluorescent Tagged Glucose Bioprobes and their Facilitation of the Drug Discovery Process
Fluorescent tagged glucose probes offer an attractive alternative to traditional, radioactive based methods for measuring glucose flux in biological systems. Thus, it could be envisaged that these probes would be widely used. However, this is not the case and, since their development in the mid-1980s, fluorescent tagged glucose bioprobes are relatively underutilized in biological research compared to radioactive methods, with only a small number (<10) publications per year using these probes. However, within the past five years there has been a surge in research activity. By the year 2012, numerous novel probes were developed and the number of research publications dramatically increased. This was especially relevant for drug discovery applications related to cancer, neurology and diabetes research. In this review article, we discuss the research impact of these bioprobes and assess which probes have been most successfully applied to drug discovery applications. Significantly, we also discuss latest research that shows the potential of these probes to be used for drug discovery in animal models and their application to in vivo-based drug validation. Overall, we hope that this review will raise awareness of the research opportunities that these probes offer to the drug discovery research community.


A RheuMetric physician checklist to quantitate levels of inflammation, damage and distress on 0-10 visual analogue scales.
A physician global assessment of patient status (DOCGL) was designed initially to quantitate inflammatory activity in rheumatoid arthritis (RA) clinical trials, in which patients are selected for high levels of activity. However, in patients seen in routine care with various diagnoses, and even in some RA patients selected for clinical trials, DOCGL also may be affected by joint damage and/or patient distress. To clarify DOCGL on a 0-10 visual analogue scale (VAS), 3 additional 0-10 VAS have been developed to record physician estimates of inflammation (DOCINF), damage (DOCDAM), and distress (DOCSTR) (such as fibromyalgia (FM)/depression). Results from 3 locales for these 4 VASs are summarised, including 478 initial-visit patients from Tennessee in 1996 to 2007, 197 initial-visit patients from Pennsylvania in 2008 to 2012, and a random visit of 739 patients from Illinois in 2014 to 2015. Highest DOCGL estimates were seen at the 3 sites in FM, followed by RA and osteoarthritis (OA), spondyloarthropathies (SpA), gout, and systemic lupus erythematosus (SLE). Highest DOCINF (inflammation) estimates were seen in RA and SpA, followed by gout, SLE, FM, and OA. Highest DOCDAM (damage) estimates were in OA, followed by RA, SpA, gout, SLE and FM. Highest DOCSTR (distress) estimates were in FM, followed by OA, RA, SpA, SLE, and gout. In the 2 earlier series, DOCDAM was considerably higher than DOCINF only in OA, and lower in the other diagnoses, although within 50% of DOCINF. In more recent patients from Illinois, mean DOCDAM was higher than DOCINF in all 6 diagnoses. The 0-10 physician VASs depict the expertise of a rheumatologist to distinguish between inflammation, damage and distress in an individual patient and rate levels as quantitative data beyond narrative descriptions. These VASs appear informative for rheumatology care, documentation, and research.


Investigation of impact and spreading of molten nanosized gold droplets on solid surfaces.
Understanding the impact dynamics and spreading of molten nanosized droplets on a solid surface is a crucial step towards the design and control of nano-fabrication in many novel applications of nanotechnology. In this context, molecular dynamic (MD) simulations have been conducted to compute temperature and dynamic contact angles of nano-droplets during impact. The evolution of the morphology of a molten metallic nano-droplet impacting on a substrate has been studied using a combination of experimental and simulation techniques. Femtosecond lasers have been used to transfer nanosized gold droplets. Droplet morphology calculated in MD simulations is found to be in good agreement with that seen in scanning electron microscopy (SEM) images. It is found that the spreading of nanoscale molten gold droplets upon impact is enhanced by increasing the droplet impact energy. As observed in experimental data, MD simulation results show that a high droplet-substrate heat transfer rate together with increased wettability of the substrate facilitates spreading and results in a thinner metal deposit after solidification.


In vitro dentin permeability after application of Gluma® desensitizer as aqueous solution or aqueous fumed silica dispersion.
To assess and to compare the effects of Gluma® Desensitizer (GDL) with an experimental glutaraldehyde and HEMA containing fumed silica dispersion (GDG) on dentin permeability using a chemiluminous tracer penetration test. Twenty disc-shaped dentin specimens were dissected from extracted human third molars. The dentin specimens were mounted in a split chamber device for determination of permeability under liquid pressure using a photochemical method. Ten specimens were randomly selected and allocated to the evaluation groups Gluma® Desensitizer as aqueous solution and glutaraldehyde/HEMA as fumed silica dispersion, respectively. Dentin disc permeability was determined at two pressure levels after removal of smear with EDTA, after albumin soaking, and after application of the desensitizing agents. Two desensitizer-treated and rinsed specimens of each group were examined by scanning electron microscopy (SEM) for surface remnants. Comparatively large standard deviations of the mean EDTA reference and albumin soaked samples permeability values reflected the differences of the dentin substrates. The mean chemiluminescence values of specimen treated with GDL and GDG, respectively, were significantly reduced after topical application of the desensitizing agents on albumin-soaked dentin. The effects of GDL and GDG on permeability were not significantly different. Treated specimens showed no surface remnants after rinsing. The experimental desensitizer gel formulation reduced dentin permeability as effectively as the original Gluma® Desensitizer solution.


Prevalence and variability of current depressive disorder in 27 European countries: a population-based study.
We aimed to estimate the prevalence of current depressive disorder in 27 European countries, and to explore differences in prevalence between European countries and by gender. In this population-based study, we analysed data from respondents living in 27 European countries who were included in the second wave of the European Health Interview Survey, collected between 2013 and 2015. We assessed the prevalence of current depressive disorder using the eight-item Patient Health Questionnaire (PHQ-8), with depressive disorder defined as a PHQ-8 score of 10 or higher. Prevalence estimates and 95% CIs were calculated for all 27 countries overall and for each country individually. We assessed variation in prevalence (country vs the rest of Europe) using crude and adjusted prevalence ratios obtained from negative binomial regression models. We did all analyses for the total sample and stratified by gender. Our analysis sample comprised 258 888 individuals, of whom 117 310 (weighted proportion 47·8%) were men and 141 578 (52·2%) were women. The overall prevalence of current depressive disorder was 6·38% (95% CI 6·24-6·52) with important variation across countries, ranging from 2·58% (2·14-3·02) in the Czech Republic to 10·33% (9·33-11·32) in Iceland. Prevalence was higher in women (7·74% [7·53-7·95]) than in men (4·89% [4·71-5·08]), with clear gender differences for all countries except Finland and Croatia. Compared with the other European countries in our sample, those with the highest adjusted prevalence ratios were Germany (1·80 [1·71-1·89]) and Luxembourg (1·50 [1·35-1·66]), and those with the lowest adjusted prevalence ratios were Slovakia (0·28 [0·24-0·33]) and the Czech Republic (0·32 [0·27-0·38]). Depressive disorders, although common across Europe, vary substantially in prevalence between countries. These results could be a baseline for monitoring the prevalence of current depressive disorder both at a country level in Europe and for planning health-care resources and services. UK Medical Research Council and CIBER Epidemiology and Public Health (CIBERESP).


Copper absorption, excretion, and retention by young men consuming low dietary copper determined by using the stable isotope 65Cu.
A study was conducted in young men to evaluate the effect of a low-copper diet on copper absorption, excretion, and retention. Eleven young men were confined to a metabolic research unit for 90 d. The study was divided into three periods, with dietary copper as the only variable. Dietary copper intake was 0.66 mg/d for 24 d, 0.38 mg/d for 42 d, and 2.49 mg/d for 24 d. The stable isotope 65Cu was fed to five of the subjects once during the first and last dietary period and twice, early and late, in the second period to determine copper absorption. 65Cu was infused into an arm vein of the other six subjects once during each dietary period to estimate excretion of endogenous copper. Total copper and 65Cu were determined by isotope dilution with thermal-ionization mass spectrometry. Fractional absorption was significantly higher during the low-copper period than in either period with higher dietary copper and excretion of the infused isotope was significantly lower in the low-copper period. Subjects were in negative balance early in the first two periods but achieved balance by the end of those periods. They retained copper during the highest dietary copper period (third period). The results suggest that endogenous copper excretion is a major point of regulation of the body's copper stores. Regulation of absorption and of endogenous excretion in response to dietary copper intake helps to protect against deficiency and toxicity. However, this regulation was not sufficient to maintain copper status at the lowest intake of dietary copper, 0.38 mg/d.


Diminished A-LAK cytotoxicity and proliferation accompany disease progression in chronic myelogenous leukemia.
We have compared the proliferative and cytotoxic capacities of a highly purified population of recombinant interleukin-2 (rIL-2)-activated peripheral blood mononuclear cells (PBMNC), termed adherent lymphokine-activated killer cells (A-LAK), in 15 chronic phase (CP) and 10 advanced disease (AD) Ph-positive chronic myelogenous leukemia (CML) patients. The selective enrichment of CML A-LAK cells depended on their propensity to adhere to plastic and to proliferate when cultured in the presence of rIL-2 for 14 days. In both CP and AD patients, 14-day culture resulted in growth of a uniform population of large granular lymphocytes. While less than 10% of the A-LAK cells were CD56-/CD3+ (mature T lymphocytes), 82% +/- 12% of A-LAK cells from early CP patients (diagnosed less than 1 year from study), 84% +/- 3% of A-LAK cells from late CP patients (studied greater than 1 year after diagnosis), and 87% +/- 3% of A-LAK cells from AD patients were CD56+/CD3- (activated natural killer [NK] cells). No bcr gene rearrangement could be found in A-LAK cells from 13 CP and six AD CML patients studied. A-LAK cells from seven early CP CML patients displayed similar cytotoxicity against K562 (80% +/- 7% lysis at effector:target ratio of 20:1) and against Raji (80% +/- 12% lysis) compared with A-LAK from 17 normal individuals (72% +/- 3% K562 lysis, P = .21; 74% +/- 5% Raji lysis, P = .39). However, the cytotoxicity of A-LAK cells from eight late CP patients (59% +/- 5% K562 lysis, P = .02; 52% +/- 8% Raji lysis, P = .02) and that of 10 AD patients studied at any point after diagnosis (31% +/- 3% K562 lysis, P less than .001; 25% +/- 6% Raji lysis, P less than .001) was significantly lower than that of seven early CP CML patients and 17 normals. The proliferative potential of A-LAK cells from seven early CP CML patients (291 +/- 191-fold) was significantly greater than that of A-LAK cells from 17 normal individuals (23 +/- 3-fold, P = .03), eight late CP patients (46 +/- 17-fold, P = .02), and 10 AD patients (5.4 +/- 1.9-fold, P = .01). In contrast to CML A-LAK, K562 cytotoxicity of unstimulated mature peripheral blood NK cells was significantly lower in early CP CML patients than in normals and remained low at all stages of disease.(ABSTRACT TRUNCATED AT 400 WORDS)


Genetically enhancing mitochondrial antioxidant activity improves muscle function in aging.
Age-related skeletal muscle dysfunction is a leading cause of morbidity that affects up to half the population aged 80 or greater. Here we tested the effects of increased mitochondrial antioxidant activity on age-dependent skeletal muscle dysfunction using transgenic mice with targeted overexpression of the human catalase gene to mitochondria (MCat mice). Aged MCat mice exhibited improved voluntary exercise, increased skeletal muscle specific force and tetanic Ca(2+) transients, decreased intracellular Ca(2+) leak and increased sarcoplasmic reticulum (SR) Ca(2+) load compared with age-matched wild type (WT) littermates. Furthermore, ryanodine receptor 1 (the sarcoplasmic reticulum Ca(2+) release channel required for skeletal muscle contraction; RyR1) from aged MCat mice was less oxidized, depleted of the channel stabilizing subunit, calstabin1, and displayed increased single channel open probability (Po). Overall, these data indicate a direct role for mitochondrial free radicals in promoting the pathological intracellular Ca(2+) leak that underlies age-dependent loss of skeletal muscle function. This study harbors implications for the development of novel therapeutic strategies, including mitochondria-targeted antioxidants for treatment of mitochondrial myopathies and other healthspan-limiting disorders.


Penetration of nicotine and nitrosonornicotine across porcine oral mucosa.
The use of smokeless tobacco (chewing tobacco or snuff) results in contact between this material and the lining of the mouth over relatively long periods of time. As tobacco contains nicotine and a carcinogen, nitrosonornicotine (NNN) there is a likelihood of systemic absorption of both these compounds under these conditions. This study measured the penetration of nicotine and NNN across skin and various regions of oral mucosa. Specimens of porcine skin, keratinized gingiva and non-keratinized floor of mouth and cheek mucosa were placed in perfusion chambers and the flux across these tissues of isotopically labeled water, nicotine and NNN was determined; permeability constants were calculated for each compound and for each region and the values compared statistically. Skin showed a lower permeability than the oral regions to all compounds and floor of mouth was generally the most permeable site. Nicotine passed across oral mucosa almost as rapidly as water, the most permeable region being floor of mouth, whereas gingiva and buccal mucosa showed a similar, but lower, permeability. The non-keratinized oral regions were most permeable to NNN and this can be related to the known distribution of oral carcinomas, regions with the higher permeability being sites that show the greatest tendency to develop such lesions.


Limits to Genomic Divergence Under Sexually Antagonistic Selection.
Since the autosomal genome is shared between the sexes, sex-specific fitness optima present an evolutionary challenge. While sexually antagonistic selection might favor different alleles within females and males, segregation randomly reassorts alleles at autosomal loci between sexes each generation. This process of homogenization during transmission thus prevents between-sex allelic divergence generated by sexually antagonistic selection from accumulating across multiple generations. However, recent empirical studies have reported high male-female FST statistics. Here, we use a population genetic model to evaluate whether these observations could plausibly be produced by sexually antagonistic selection. To do this, we use both a single-locus model with nonrandom mate choice, and individual-based simulations to study the relationship between strength of selection, degree of between-sex divergence, and the associated genetic load. We show that selection must be exceptionally strong to create measurable divergence between the sexes and that the decrease in population fitness due to this process is correspondingly high. Individual-based simulations with selection genome-wide recapitulate these patterns and indicate that small sample sizes and sampling variance can easily generate substantial male-female divergence. We therefore conclude that caution should be taken when interpreting autosomal allelic differentiation between the sexes.


Cardiac troponin I elevation in hospitalized patients without acute coronary syndromes.
Increase of cardiac troponins occurs in a variety of clinical situations in the absence of an acute coronary syndrome (ACS). Few data exist regarding the incidence, clinical characteristics, and predictive value of various cardiac diagnostic tests and outcome of patients with a non-ACS-related troponin increase. We studied 883 consecutive hospitalized patients with increased cardiac troponin I levels. The discharge diagnosis was reclassified and troponin increase attributed to ACS or another process. Clinical data and results of cardiac diagnostic tests were collected. Patients were followed for a median of 30 months. Three hundred eleven patients were classified as having a non-ACS-related troponin increase (35.2%). An alternative explanation for troponin increase was found in 99% of these patients. Troponin level had poor accuracy in discriminating patients with and without ACS (area under the receiver operating characteristics curve 0.63). Coronary angiography was frequently unhelpful in excluding a non-ACS-related troponin increase because 77% of patients in the non-ACS group had significant flow-limiting coronary artery disease. Patients with non-ACS-related troponin increase had significantly higher in-hospital (hazard ratio 2.8, 95% confidence interval 2.0 to 3.8) and long-term (hazard ratio 2.0, 95% confidence interval 1.6 to 2.5) mortalities compared with patients with ACS. In conclusion, cardiac troponin level is frequently increased in hospitalized patients in the absence of an ACS and portends poor short- and long-term outcomes. Most of these patients have an alternative explanation for cardiac troponin increase. Cardiac diagnostic procedures are frequently unhelpful in excluding a non-ACS-related troponin increase.


Repetitive DNA in the Architecture, Repatterning, and Diversification of the Genome of Aegilops speltoides Tausch (Poaceae, Triticeae).
The genome's adaptability to environmental changes, especially during rapid climatic fluctuations, underlies the existence and evolution of species. In the wild, genetic and epigenetic genomic changes are accompanied by significant alterations in the complex nuclear repetitive DNA fraction. Current intraspecific polymorphism of repetitive DNA is closely related to ongoing chromosomal rearrangements, which typically result from erroneous DNA repair and recombination. In this study, we addressed tandem repeat patterns and interaction/reshuffling both in pollen mother cell (PMC) development and somatogenesis in the wild diploid cereal Aegilops speltoides, with a focus on genome repatterning and stabilization. Individual contrasting genotypes were investigated using the fluorescent in situ hybridization (FISH) approach by applying correlative fluorescence and electron microscopy. Species-specific Spelt1 and tribe-specific Spelt52 tandem repeats were used as the markers for monitoring somatic and meiotic chromosomal interactions and dynamics in somatic interphase nuclei. We found that, the number of tandem repeat clusters in nuclei is usually lower than the number on chromosomes due to the associations of clusters of the same type in common blocks. In addition, tightly associated Spelt1-Spelt52 clusters were revealed in different genotypes. The frequencies of nonhomologous/ectopic associations between tandem repeat clusters were revealed in a genotype-/population-specific manner. An increase in the number of tandem repeat clusters in the genome causes an increase in the frequencies of their associations. The distal/terminal regions of homologous chromosomes are separated in nuclear space, and nonhomologous chromosomes are often involved in somatic recombination as seen by frequently formed interchromosomal chromatin bridges. In both microgametogenesis and somatogenesis, inter- and intrachromosomal associations are likely to lead to DNA breaks during chromosome disjunction in the anaphase stage. Uncondensed/improperly packed DNA fibers, mainly in heterochromatic regions, were revealed in both the meiotic and somatic prophases that might be a result of broken associations. Altogether, the data obtained showed that intraorganismal dynamics of repetitive DNA under the conditions of natural out-crossing and artificial intraspecific hybridization mirrors the structural plasticity of the Ae. speltoides genome, which is interlinked with genetic diversity through the species distribution area in contrasting ecogeographical environments in and around the Fertile Crescent.


Secondary diurnal enuresis: evaluation of cueing and reinforcement interventions with a sensory-impaired youth.
Secondary enuresis consists of sporadic urinary accidents in persons who, otherwise, possess toileting skills. The present study evaluated the behavioural treatment of secondary enuresis displayed by a 19-year-old, developmentally disabled female with multiple sensory impairments. Treatment consisted of external and self-cueing procedures combined with reinforcement for on-toilet urination. As evaluated in a multiple baseline design, intervention resulted in a decrease in wetting incidents and an increase in self-initiated toileting. Treatment effects were maintained through a 7-month follow-up assessment.


More than Meets the Eye: Orbital Swelling in an Adolescent with Sickle Cell Disease.
Periorbital swelling is a clinical presentation with a broad differential and potentially deleterious consequence. Causes range from benign, including allergic reaction, to vision- and life-threatening, including orbital cellulitis and orbital infarction. The recent climate of SARS-CoV-2 has further complicated this differential, as the virus poses broad clinical presentations with new manifestations reported frequently. Rapid identification of the underlying etiology is crucial, as treatment approaches diverge greatly. Here, we report the case of an African American adolescent male with a history of homozygous sickle cell anemia presenting to an inner city hospital with bilateral periorbital swelling amid the coronavirus pandemic. Differentials, including orbital cellulitis, COVID-MIS-C, orbital inflammatory syndrome, Hoagland sign, and orbital infarction secondary to sickle cell crisis are contrasted. We contrast our case with 12 case reports of orbital infarction in the setting of sickle cell crisis within the past 10 years, highlighting how these presentations, along with commonly reported findings of orbital infarction, compare with our patient.


Exploring the clinical utility of two staging models for bipolar disorder.
To assess the clinical utility of two staging models for bipolar disorder by examining distribution, correlation, and the relationship to external criteria. These are primarily defined by the recurrence of mood episodes (model A), or by intra-episodic functioning (model B). In the Dutch Bipolar Cohort, stages according to models A and B were assigned to all patients with bipolar-I-disorder (BD-I; N = 1396). The dispersion of subjects over the stages was assessed and the association between the two models calculated. For both models, change in several clinical markers were concordant with the stage was investigated. Staging was possible in 87% of subjects for model A and 75% for model B. For model A, 1079 participants (93%) were assigned to stage 3c (recurrent episodes). Subdividing stage 3c with cut-offs at 5 and 10 episodes resulted in subgroups containing 242, 510, and 327 subjects. For model B, most participants were assigned to stage II (intra-episodic symptoms, N = 431 (41%)) or stage III (inability to work, N = 451 (43%)). A low association between models was found. For both models, the clinical markers "age at onset," "treatment resistance," and "episode acceleration" changed concordant with the stages. The majority of patients with BD-I clustered in recurrent stage 3 of Model A. Model B showed a larger dispersion. The stepwise change in several clinical markers supports the construct validity of both models. Combining the two staging models and sub-differentiating the recurrent stage into categories with cut-offs at 5 and 10 lifetime episodes improves the clinical utility of staging for individual patients.