Separation of viable T and B lymphocytes using a cytochemical stain, Hoechst 33342.
Data are presented that show that a histochemical stain, Hoeschst 33342, can be used to discriminate between viable B and T lymphocytes in the mouse. Quantitative analysis of the staining of cells from various lymphoid tissues with Hoechst 33342 using a Fluorescence-Activated Cell Sorter (FACS) indicates that two populations of cells can be identified. In the spleen approximately 60% of the lymphocytes can be classified as brightly stained with 1 microgram/ml of Hoechst 33342, while in the lymph node only 40% of the cells stain brightly. Thymocytes exhibit only the dull staining profile. Separation of these two populations from the spleen using the FACS and reanalyzing them for cell surface antigenic markers shows that the lymphocytes stained brightly with Hoechst 33342 are predominantly immunoglobulin positive, while the cells that stain less brightly express Thy 1.2. This indicates that a histochemical stain correlates directly with classical immunological markers on cell surfaces.


Alkenyl Isocyanide Conjugate Additions: A Rapid Route to γ-Carbolines.
Isocyanides are exceptional building blocks, the wide deployment of which in multicomponent and metal-insertion reactions belies their limited availability. The first conjugate addition/alkylation to alkenyl isocyanides is described, which addresses this deficiency. An array of organolithiums, magnesiates, enolates, and metalated nitriles add conjugately to β- and β,β-disubstituted arylsulfonyl alkenyl isocyanides to rapidly assemble diverse isocyanide scaffolds. The intermediate metalated isocyanides are efficiently trapped with electrophiles to generate substituted isocyanides incorporating contiguous tri- and tetra-substituted centers. The substituted isocyanides are ideally functionalized for elaboration into synthetic targets as illustrated by the three-step synthesis of γ-carboline N-methyl ingenine B.


Haemodynamics of pelvic nerve induced penile erection in the dog: possible mediation by vasoactive intestinal polypeptide.
The haemodynamics of erection were elucidated in the anaesthetized dog by analysing in quantitative terms the changes of penile arterial inflow, venous outflow and tissue volume during graded pelvic nerve stimulation. The study also provides information on possible neurotransmitter mechanisms of the erectile response. Erection evoked by pelvic nerve stimulation appeared to result from two main circulatory events: first, there was a prompt dilatation of the penile 'resistance vessels', causing a greatly increased arterial inflow which in the early phase bypassed the cavernous bodies and, hence, increased venous outflow to the same extent. Secondly, the erectile response proper began after a distinct delay (approximately equal to 20 s). This was apparently caused by sudden opening of low resistance 'shunt vessels' diverting part of the arterial inflow into the cavernous bodies, leading to rapid filling. During the filling phase arterial inflow greatly exceeded venous outflow, and returned to the venous outflow level again in the steady state of full erection. The initial dilator response seemed to ensure rapid erection by establishing a high pressure head from the arterial microvessels to the cavernous spaces. The threshold frequency for the penile vasodilator response to pelvic nerve stimulation was 1-2 Hz and was always higher for the erectile volume response, viz. 2-4 Hz. Maximal effects for both were obtained at 16 Hz, causing on the average a 25-fold increase in peak arterial inflow, a 17-fold increase in venous outflow and a 107% increase in penile volume. Muscarinic blockade by atropine caused no significant decrease in the blood flow response induced by pelvic nerve stimulation, but clearly curtailed the erectile response. This indicates that the dilatation of the penile 'resistance vessels' is mainly non-cholinergic in nature, whereas a cholinergic mechanism seems to contribute to the erectile volume response proper. Pelvic nerve stimulation caused a substantial output of vasoactive intestinal polypeptide (VIP) from the penis which was correlated in onset and duration to the vasodilator response. Intra-arterial (I.A.) infusion of VIP elicited moderate erection and a penile vasodilator response which resembled the neural response. Similar effects were evoked by I.A. infusion of substance P, but the output of this peptide from the penis during stimulation was poorly correlated to the vascular events. These in vivo observations indicate that VIP might be the neurotransmitter responsible for the non-cholinergic pelvic nerve induced penile vasodilatation.


[MRI-based radiotherapy planning].
MRI-based radiotherapy planning is a topical subject due to the introduction of a new generation of treatment machines combining a linear accelerator and a MRI. One of the issues for introducing MRI in this task is the lack of information to provide tissue density information required for dose calculation. To cope with this issue, two strategies may be distinguished from the literature. Either a synthetic CT scan is generated from the MRI to plan the dose, or a dose is generated from the MRI based on physical underpinnings. Within the first group, three approaches appear: bulk density mapping assign a homogeneous density to different volumes of interest manually defined on a patient MRI; machine learning-based approaches model local relationship between CT and MRI image intensities from multiple data, then applying the model to a new MRI; atlas-based approaches use a co-registered training data set (CT-MRI) which are registered to a new MRI to create a pseudo CT from spatial correspondences in a final fusion step. Within the second group, physics-based approaches aim at computing the dose directly from the hydrogen contained within the tissues, quantified by MRI. Excepting the physics approach, all these methods generate a synthetic CT called "pseudo CT", on which radiotherapy planning will be finally realized. This literature review shows that atlas- and machine learning-based approaches appear more accurate dosimetrically. Bulk density approaches are not appropriate for bone localization. The fastest methods are machine learning and the slowest are atlas-based approaches. The less automatized are bulk density assignation methods. The physical approaches appear very promising methods. Finally, the validation of these methods is crucial for a clinical practice, in particular in the perspective of adaptive radiotherapy delivered by a linear accelerator combined with an MRI scanner.


Erythema ab igne: evolving technology, evolving presentation.
We present a case of a 49-year-old woman with erythema ab igne on her posterior thighs owing to 2-4 hours per day of seat heater use in her car. Erythema ab igne is caused by prolonged exposure to a heat source. It used to be caused mainly by wood stoves used to heat homes. Erythema ab igne is now more often related to other heat sources, including heating pads, laptop computers, and car seat heaters, as in our case. As technology changes, so does the presentation of skin conditions that are related to technology.


Explaining variation in ventral and inguinal hernia repair outcomes: A population-based analysis.
No study has evaluated the relative importance of patient, surgeon, and hospital-level factors on surgeon and hospital variation in hernia reoperation rates. This population-based retrospective cohort study evaluated factors associated with variation in reoperation rates for recurrence after initial ventral hernia repair and inguinal hernia repair. The Statewide Planning and Research Cooperative System identified initial ventral hernia repairs and inguinal hernia repairs in New York state from 2003-2009. Mixed-effects Cox proportional hazards analyses were performed assessing factors associated with surgeon/hospital variation in 5-year reoperation rates for hernia recurrence. Among 78,267 ventral hernia repairs and 124,416 inguinal hernia repairs, the proportion of total variation in reoperation rates attributable to individual surgeons compared with hospitals was 87% for ventral hernia repairs and 92% for inguinal hernia repairs. In explaining variation in ventral hernia repair reoperation between surgeons, 19% was attributable to patient-level factors, 4% attributable to mesh placement, and 10% attributable to surgeon volume and type of board certification. In explaining variation in inguinal hernia repair reoperation between surgeons, 1.1% was attributable to mesh placement and 10% was attributable to surgeon volume and years of experience. However, 67% of the variation between surgeons for ventral hernia repair and 89% of the variation between surgeons for inguinal hernia repair remained unexplained by factors in the models. The majority of variation in hernia reoperation rates is attributable to surgeon-level variation. This suggests that hernia recurrence may be an appropriate surgeon quality metric. While modifiable factors such as mesh placement and surgeon characteristics play roles in surgeon variation, future research should focus on identifying additional surgeon attributes responsible for this variation.


[Therapy of bone and joint changes in Albers-Schönberg osteopetrosis].
The history, genetic heterogenity, clinical appearance, radiologic morphology and specific pathology of Albers-Schönberg disease are described, with special reference to its orthopedic aspects, on the basis of 14 case histories. The orthopedic problems (osteomyelitis, delayed fracture and osteotomy healing, growth deviations of long bones, spinal pathology, arthrosis and fatigue fractures) are described in detail and documented. The disease is often recognized only by chance when routine X-ray examinations are performed. More serious manifestations of the disease, however, such as spontaneous osteomyelitis, fatigue fractures, coxarthrosis or gonarthrosis with or without growth aberrations, pose difficult problems of treatment. Delayed callus formation is routinely observed; this demands special osteosynthesis techniques must be constantly borne in mind during and after operative care.


Atypical Presentation of Systemic Arthritis in a Toddler with Down Syndrome.
Systemic juvenile idiopathic arthritis (sJIA) is a chronic, inflammatory disease of childhood, which is characterized by the combination of arthritis, serositis, daily, high-spiking fevers, and evanescent macular rash and can present with the life-threatening complication of macrophage activation syndrome (MAS). Children with Down syndrome (DS) have complex medical challenges related to abnormalities in their immune system, which can cause a broad spectrum of disease manifestations, which can occur atypically. Children with DS are at increased risk for arthritis and interstitial lung disease (ILD) associated with sJIA that has high mortality. This case report outlines an atypical presentation of sJIA in a 21-month-old male with DS in which fever was not part of the initial presentation of sJIA and then later developed MAS and ILD. Due to broad spectrum of disease and atypical presentation in children with DS, this case report was created to increase awareness of atypical presentations of rheumatic disease in children with DS.


Oocytes and hypoxanthine orchestrate the G2-M switch mechanism in ovarian granulosa cells.
In mammalian growing follicles, oocytes are arrested at the diplotene stage (which resembles the G2/M boundary in mitosis), while the granulosa cells (GCs) continue to proliferate during follicular development, reflecting a cell cycle asynchrony between oocytes and GCs. Hypoxanthine (Hx), a purine present in the follicular fluid, has been shown to induce oocytes meiotic arrest, although its role in GC proliferation remains ill-defined. Here, we demonstrate that Hx indiscriminately prevents G2-to-M phase transition in porcine GCs. However, oocyte-derived paracrine factors (ODPFs), particularly GDF9 and BMP15, maintain the proliferation of GCs, partly by activating the ERK1/2 signaling and enabling the G2/M transition that is suppressed by Hx. Interestingly, GCs with lower expression of GDF9/BMP15 receptors appear to be more sensitive to Hx-induced G2/M arrest and become easily detached from the follicular wall. Importantly, Hx-mediated inhibition of G2/M progression instigates GC apoptosis, which is ameliorated in the presence of GDF9 and/or BMP15. Therefore, our data indicate that the counterbalance of intrafollicular factors, particularly Hx and oocyte-derived GDF9/BMP15, fine-tunes the development of porcine follicles by regulating the cell cycle progression of GCs.


Differential susceptibility of naive and memory CD4+ T cells to the cytopathic effects of infection with human immunodeficiency virus type 1 strain LAI.
CD4+ T lymphocytes of individuals infected with human immunodeficiency virus type 1 (HIV-1) exhibit a qualitative defect in their ability to mount memory responses to previously encountered antigens although their responses to mitogens remain normal. T cells responsible for memory responses can be distinguished from naive T cells based on differential expression of isoforms of the tyrosine phosphatase CD45. It has been suggested that memory CD4+ T cells from infected individuals have a greater virus burden than naive CD4+ T cells and that this accounts for the loss of recall responses in infected individuals. However, it has been unclear whether naive and memory T cells are equally susceptible to infection and to the cytopathic effects of the virus. We therefore infected highly purified resting naive and memory CD4+ T cells from HIV-1-seronegative individuals with HIV-1(LAI). Infected cells were then stimulated with phytohemagglutinin to render them permissive for viral replication. Cell viability and growth rate were monitored for 8 to 10 days as indicators of cytopathic effects induced by HIV-1(LAI). Our results indicated that naive and memory CD4+ T cells display marked differences in susceptibility to the cytopathic effects induced by HIV-1(LAI), infection. The cytopathic effects induced by HIV-1(LAI) were much more severe in memory CD4+ T cells than in naive CD4+ T cells. Differential cytopathic effects in naive and memory T cells were not due to differences in virus entry into and replication in these cell populations. Rather, memory cells were more susceptible to cytopathic effects. Pronounced cytopathic effects in memory cells were clearly detectable at 7 day postinfection. Cell death occurred at the single-cell level and was not accompanied by syncytium formation. The growth rate of infected memory CD4+ T cells was also severely compromised compared to that of naive CD4+ T cells, whereas the growth rates of both uninfected naive and memory CD4+ T cells were approximately the same. At least a portion of the dying cells exhibited biochemical changes characteristic of apoptosis. These results suggest that the selective functional defects present in the memory CD4+ T-cell subset of HIV-1-infected individuals may in part be the result of the greater susceptibility of memory T cells to cytopathic effects induced by HIV-1.


Cyclosporine A drives a Th17- and Th2-mediated posttransplant obliterative airway disease.
Calcineurin-inhibitor refractory bronchiolitis obliterans (BO) represents the leading cause of late graft failure after lung transplantation. T helper (Th)2 and Th17 lymphocytes have been associated with BO development. Taking advantage of a fully allogeneic trachea transplantation model in mice, we addressed the pathogenicity of Th cells in obliterative airway disease (OAD) occurring in cyclosporine A (CsA)-treated recipients. We found that CsA prevented CD8(+) T cell infiltration into the graft and downregulated the Th1 response but affected neither Th2 nor Th17 responses in vivo. In secondary mixed lymphocyte cultures, CsA dramatically decreased donor-specific IFN-γ production, enhanced IL-17 production and did not affect IL-13. As CD4(+) depletion efficiently prevented OAD in CsA-treated recipients, we further explored the role of Th2 and Th17 immunity in vivo. Although IL-4 and IL-17 deficient untreated mice developed an OAD comparable to wild-type recipients, a single cytokine deficiency afforded significant protection in CsA-treated recipients. In conclusion, CsA treatment unbalances T helper alloreactivity and favors Th2 and Th17 as coexisting pathways mediating chronic rejection of heterotopic tracheal allografts.


Islet cell tumor.
A case of islet cell tumor occurring in a patient with the multiple endocrine neoplasia type I syndrome is reported. Immunostaining for insulin was strongly positive in the tumor cells. Numerous dense-core granules of endocrine caliber were identified ultrastructurally. Morphometric analysis of the secretory granules in 20 islet cell tumors gave a granule size of 182 +/- 52 nm (mean +/- standard deviation).


The Effect of Two Acute Bouts of Exercise on Oxidative Stress, Hematological, and Biochemical Parameters, and Rectal Temperature in Trained Canicross Dogs.
Canicross is a sport discipline that connects human and canine athletes in running. Changes in physiological, hematological, and biochemical parameters, and exercise-induced oxidative stress have not been thoroughly characterized in canicross dogs. The aim of our study was the assessment of the health status of trained canicross dogs that were subjected to two acute bouts of exercise with their owners during the training season. Health status was assessed by measuring the rectal temperature, hematological and biochemical parameters, as well as blood oxidative stress parameters (plasma malondialdehyde, lipid peroxidation marker; whole blood glutathione peroxidase and erythrocyte superoxide dismutase1, antioxidant enzymes) before and during a two-day canicross training session and after a 24-h rest period. Seven trained canicross dogs (three females/four males) aged 12-120 months were included in the study. Blood samples were collected before and immediately after the first acute bout of exercise (day 1), after the second acute bout of exercise (day 2), and after 24 h of rest (day 3). Rectal temperature was measured at the same time as blood sample collection. The majority of hematological and biochemical parameters remained within reference ranges at all sampling times. Rectal temperature was significantly higher after training on days 1 and 2 compared to resting temperature on day 3. Hematological parameters did not change significantly; however, there were significant differences in urea, creatinine, creatine kinase, and triglycerides between specific sampling times. Despite significant changes, these biochemical parameters remained within reference ranges. Significant changes in biochemical parameters seem to reflect the dogs' physiological response to each acute bout of exercise, considering all biochemical parameters and rectal temperature returned to pre-exercise values after a 24-h rest period (day 3). No significant differences in oxidative stress parameters were found between any sampling times. Relatively high erythrocyte superoxide dismutase1 activity at all sampling times may indicate that the canicross dogs are adapted to training by an increased expression of antioxidant enzymes. Based on our results, we can conclude that the trained canicross dogs included in our study were healthy, in good physical condition, and fit for the two acute bouts of field exercise.


Diabetes and factors associated with cognitive and functional decline. The screening for CKD among older people across Europe (SCOPE) study.
Type 2 diabetes mellitus (DM) in older people is a heterogeneous condition that exhibits differential characteristics in comparison with younger adults. DM increases the risk of disability, is associated with dementia and loss of function, and cognition may often be interrelated and more pronounced in older patients with DM than in those without. Our aim was to evaluate the incidence of functional and/or cognitive impairment in older adults with and without DM, and its associated factors in DM participants. A 2-year prospective analysis was conducted in a European multicenter prospective cohort (SCOPE study). Older community-dwelling adults (aged ≥ 75 years) underwent a comprehensive geriatric assessment. New functional and/or cognitive decline was explored. Of 1611 participants, 335 (22.0%) had DM at baseline. The percentage of participants scoring at least one ADL impairment and/or cognitive impairment (MMSE < 24) was similar in both groups (9.6%). Factors associated with any new disability in participants with DM in the multivariate analysis were female sex (OR 3.28, 95% CI 1.42-7.56), history of stroke (OR 4.58, 95% CI 1.64-12.7), and greater IADL dependency (OR 1.08 95% CI 1.02-1.15). Association between DM and cognitive or functional decline in outpatients of 75 years and older was not found, but factors such as female gender, history of stroke, and IADL dependency could be related. Decline in functional and cognitive status of community-dwelling older adults with DM was similar to participants without DM in a short period of 2 years of follow-up, though several clinical factors may increase its risk in this population.


Phylogeographic analysis elucidates the influence of the ice ages on the disjunct distribution of relict dragonflies in Asia.
Unusual biogeographic patterns of closely related groups reflect events in the past, and molecular analyses can help to elucidate these events. While ample research on the origin of disjunct distributions of different organism groups in the Western Paleartic has been conducted, such studies are rare for Eastern Palearctic organisms. In this paper we present a phylogeographic analysis of the disjunct distribution pattern of the extant species of the strongly cool-adapted Epiophlebia dragonflies from Asia. We investigated sequences of the usually more conserved 18 S rDNA and 28 S rDNA genes and the more variable sequences of ITS1, ITS2 and CO2 of all three currently recognised Epiophlebia species and of a sample of other odonatan species. In all genes investigated the degrees of similarity between species of Epiophlebia are very high and resemble those otherwise found between different populations of the same species in Odonata. This indicates that substantial gene transfer between these populations occurred in the comparatively recent past. Our analyses imply a wide distribution of the ancestor of extant Epiophlebia in Southeast Asia during the last ice age, when suitable habitats were more common. During the following warming phase, its range contracted, resulting in the current disjunct distribution. Given the strong sensitivity of these species to climatic parameters, the current trend to increasing global temperatures will further reduce acceptable habitats and seriously threaten the existences of these last representatives of an ancient group of Odonata.


The manganese superoxide dismutase gene of Drosophila: structure, expression, and evidence for regulation by MAP kinase.
The gene encoding manganese superoxide dismutase (MnSOD) from Drosophila melanogaster has been isolated and its expression has been studied. In contrast to several mammalian MnSOD genes, the Drosophila gene contains a single intron and is transcribed into a single 0.8-kb transcript. Whole-mount in situ hybridization reveals extensive transcript accumulation in ovarian nurse cells and a heavy maternal contribution to the early embryo. Larval imaginal discs are enriched with MnSOD transcripts relative to other larval tissues, further suggesting a possible relationship between high MnSOD expression and mitotic activity. The 5'-upstream region contains several well-known regulatory elements including metal response, antioxidant response, and xenobiotic response elements (MRE, ARE, and XRE, respectively), sites for activator protein-1 (AP-1), nuclear factor-kappaB (NF-kappaB), adenosine 3',5'-cyclic monophosphate regulator binding element factor (CREB), as well as classic TATA and CAAT boxes. That MnSOD expression in Drosophila is regulated in part by the transcription factor AP-1 via the MAP kinase signal transduction pathway is suggested by experiments which show that a hypomorphic mutation of the MAP kinase-encoding rolled gene substantially reduces levels of MnSOD transcripts and correlates with reduced resistance to oxidative stress in rolled mutants.


Appreciation and emotion: theoretical reflections on the MacArthur Treatment Competence Study.
When emotions are mentioned in the literature on mental competence, it is generally because they are thought to influence competence negatively; that is, they are thought to impede or compromise the cognitive capacities that are taken to underlie competence. The purpose of the present discussion is to explore the possibility that emotions might play a more positive role in the determination of competence. Using the MacArthur Treatment Competence Study as an example, it is argued that appreciation, a central theoretical concept in many contemporary approaches to competence, has important emotive components that are seldom sufficiently recognized or acknowledged. If true, this means that some leading contemporary accounts of competence need to be revised in order to make more adequate provision for the positive contribution of emotion.


The management of sinonasal inverted papilloma: our experience.
The inverted papilloma of the nose and paranasal sinuses is a controversial disease. It invades surrounded structures, shows a high recurrence rate after surgical excision and may coexist with squamous cell carcinoma. The use of the optimal surgical approach and technique is of the utmost importance, as it is related to the possibility of recurrence. In cases of tumors of limited size a conservative local excision is sufficient, while, in larger tumors filling the nasal cavity and the paranasal sinuses a more aggressive strategy is needed, such as a medial maxillectomy after lateral rhinotomy. In this paper we present our experience from treating 42 sinonasal inverted papillomas, using a surgical technique which is based on the conservation of as much part of the healthy lateral nasal wall as possible. This modified medial maxillectomy allows us to create a limited in size and normally functioning nasal cavity, thus minimizing the disadvantages of the classical medial maxillectomy without jeopardizing the safety of the operation.


Advances in proteomic techniques for biomarker discovery in COPD.
Chronic obstructive pulmonary disease (COPD) is a disorder characterized by chronic inflammation of the lung with airflow obstruction and progressive deterioration of pulmonary function. The need to discover and validate biomarkers as prognostic tools of development and progression of the disease has received further support with the advent of proteomic techniques. Liquid chromatography-mass spectrometry (LC/MS) and gel electrophoresis-mass spectrometry (2-DE/MS) have been applied to investigate the proteome of a number of lung-origin samples, including sputum, bronchoalveolar lavage fluid, exhaled-breath condensate, cells and biopsies from COPD patients. In particular, 2-DE and MS are the main proteomic approaches with 2-DE presenting the major approach for quantitative proteomics. The molecules identified as potential biomarkers of COPD may represent a preliminary step for better comprehension of the mechanisms involved in the onset/progression of the disease.


Mycoplasma gallisepticum vaccination: effects on egg transmission and egg production.
The effects of Mycoplasma gallisepticum (MG) vaccination on egg transmission of MG and egg production were evaluated. Leghorn hens vaccinated with live MG (strain F), with strain F plus MG bacterin, with one dose of MG bacterin, or with two doses of MG bacterin all transmitted MG through the egg at a significantly lower level than unvaccinated controls. Hens vaccinated with two doses of MG bacterin had the longest lag before detectable transmission of MG through the egg. All vaccinated groups were protected against the egg-production drop seen in unvaccinated hens challenged with virulent MG.


The effect of age and sex on the perception of time in life.
As a measure of their personal perception of time in life, 320 participants complete the Lines Test. Participants were asked to mark off on a line their perceived present life location between the endpoint anchors of birth and death. The percentage of the life span marked was compared with actuarial life expectancy to establish a quantitative degree of difference for each respondent. Results indicated a significant sex difference in which women across the age range investigated were more accurate as to their life location. Results also showed a significant age effect in which older participants consistently underestimated their location to a much greater degree than their younger peers. A second investigation presented an amended version of the traditional Lines Test and scaled the actuarial life span to each participant's specific age. The pattern of findings was replicated by this procedure. Reasons for this overall pattern of results are discussed in terms of what is currently understood about the perception of short intervals of time and the perception of duration across the life span.


Protective effect of vitamin C on 8-hydroxy-2'-deoxyguanosine level in peripheral blood lymphocytes of chronic hemodialysis patients.
This study focused on the effect of vitamin C on the 8-hydroxy-2'-deoxyguanosine (8-OHdG) level of cellular DNA, as well as 8-oxoguanine-DNA glycosylase 1 (hOGG1) and human MutT homologue (hMTH1) gene expression in peripheral blood lymphocytes of chronic hemodialysis patients. Sixty chronic hemodialysis patients (35 men and 25 women) were recruited to participate in a randomized, placebo-controlled study. Treatment order is block-randomized with intravenous sodium ascorbate (vitamin C, 300 mg) or placebo (0.9% saline), administered postdialysis three times a week. We evaluated 8-OHdG level, intracellular reactive oxygen species (ROS) production, and gene expression of hOGG1 and hMTH1 in peripheral blood lymphocytes by using high-performance liquid chromatography (HPLC) electrochemical detection method, flow cytometric analysis, and reverse transcription-polymerase chain reaction (RT-PCR), respectively. A total of 51 patients completed the study (26 in placebo group and 25 in vitamin C group). Mean 8-OHdG levels significantly decreased in total subjects following 8 weeks of vitamin C supplementation (22.9 vs. 18.8/10(6) dG, P < 0.01). The decrease in 8-OHdG levels after vitamin C supplementation was also noted in the patients with ferritin <500 or > or =500 microg/L and transferrin saturation (TSAT) <50 or > or =50% (P < 0.05). But 8-OHdG levels had no significant changes in total patients or in the four subgroups of patients treated with placebo as compared to their baselines. Intracellular ROS production by lymphocytes from the four subgroups of patients, either spontaneous (P < 0.05) or phorbol-12-myristate-13-acetate (PMA)-stimulated (P < 0.001), was significantly reduced after 8 weeks vitamin C supplementation. Steady-state hOGG1 mRNA levels were significantly up-regulated at 24 hours after vitamin C administration (P < 0.05), but hMTH1 mRNA levels were not. The changes in the spontaneous and PMA-stimulated ROS production, and an up-regulation of hOGG1 mRNA expression were not observed in patients treated with placebo as compared to their baselines. Vitamin C supplementation in chronic hemodialysis patients can reduce the lymphocyte 8-OHdG levels and intracellular ROS production, as well as up-regulate hOGG1 gene expression for repair. There is no compelling evidence for an in vivo pro-oxidant effect of vitamin C on lymphocyte DNA base oxidation, even in the status of increased iron stores.


Isolation of mutants of the Arabidopsis thaliana alternative oxidase (ubiquinol:oxygen oxidoreductase) resistant to salicylhydroxamic acid.
The plant-type ubiquinol:oxygen oxidoreductase, commonly called the alternative oxidase, is a respiratory enzyme thought to contain non-heme iron at its active site. To explore the structure of the enzyme by identifying amino acids involved in inhibitor-binding, a library of random mutants of the Arabidopsis thaliana alternative oxidase was constructed using error-prone polymerase chain reaction and expressed in the heme-deficient Escherichia coli SASX41B. Selection for resistance to salicylhydroxamic acid (SHAM) resulted in the recovery of four mutations. Three of these, F215L, M219I, and M219V, confer a small, but measurable resistance to SHAM of between 1.4- and 1.7-fold relative to the wild type alternative oxidase. These changes are located in a putative amphipathic helix following the second transmembrane helix. The fourth mutation, G303E, is found three residues from the C-terminus of the protein, and results in 4. 6-fold resistance to SHAM. None of the mutations have any effect on the sensitivity of the alternative oxidase to propyl gallate. The identification of distant residues involved in SHAM resistance suggests that the poorly conserved C-terminal region is in spatial proximity to the amphipathic helix, and thus located in the vicinity of the iron-binding motif.


Chemical, mineral composition, and sensory acceptability of cocoyam-based recipes enriched with cowpea flour.
The study was conducted to improve cocoyam-based recipes (steamed cocoyam paste [ebiripo], ikokore, and fried cocoyam balls [ojojo]) using different blends of cocoyam and cowpea flours (100:0, 80:20, 70:30, 60:40, and 50:50). The proximate composition, mineral composition, and sensory qualities of the recipes were determined using standard analytical procedures. The recipes had significantly (P < 0.05) higher contents of protein, fat, crude fiber, iron, zinc, sodium, and phosphorus compared with the control recipe (100% cocoyam flour). The protein content was highest in all recipes containing 50:50 cocoyam: cowpea flour (10.79%, 10.56%, 10.36% for ojojo, ikokore, and ebiripo, respectively). However, ikokore had higher iron (2.5 mg), phosphorus (92.5 mg), and zinc (1.92 mg) contents than ebiripo and ojojo. While the 80:20 recipe for ebiripo had significantly (P < 0.05) higher flavor and overall acceptability scores compared with other recipes. In conclusion, enrichment of cocoyam-based recipes with cowpea flour improved the proximate composition, mineral composition, and sensory acceptability of the foods.


A high expression of GDP-fucose transporter in hepatocellular carcinoma is a key factor for increases in fucosylation.
Changes in the levels of fucosylation regulate the biological phenotype of cancer cells and a specific fucosylation, such as fucosylated alpha-fetoprotein (AFP-L3) has been clinically used as a tumor marker for hepatocellular carcinoma (HCC). However, detailed molecular mechanisms that explain the increased fucosylation in HCC remain unknown despite 10 years of study by these researchers. Fucosylation is regulated by complicated mechanisms that involve several factors: fucosyltransferases, GDP-fucose transporter (GDP-Fuc Tr), and synthetic enzymes of GDP-fucose, such as GDP-mannose 4, 6-dehydratase (GMD), GDP-4-keto-6-deoxy-mannose-3, 5-epimerase-4-reductase (FX), and GDP-fucose pyrophosphorylase. In this study, the expression of fucosylation-related genes in HCC tissues was studied and it was found that GDP-Fuc Tr is a key factor for increases in fucosylation. A real-time reverse transcription polymerase chain reaction (RT-PCR) analysis showed significant increases in GDP-Fuc Tr and FX expression in HCC, and levels of the GMD protein were upregulated by posttranslational modification in HCC tissues. In vitro cell experiments showed that the level of GDP-Fuc Tr was the most significantly correlated with the level of cellular fucosylation and the overexpression of GDP-Fuc Tr dramatically increased fucosylation in Hep3B cells. The importance of GDP-Fuc Tr in the increase of fucosylation was also confirmed with immunohistochemical analyses. These findings suggest that the upregulation of GDP-Fuc Tr plays a pivotal role in increased fucosylation in HCC and represents an attractive target for new treatments and diagnosis for HCC.


The restoration of the vasculature of skin autografts in the rabbit.
The restoration of the vasculature of full-thickness skin autografts was investigated on the rabbit ear, the grafted discs of skin being replaced with their original orientation, or rotated 90 or 180 degrees. Circulation in the graft was first detected 1 day after grafting, increasing on the 2nd and successive days. The larger arteries in the grafts survived and became permanently joined at the periphery of the grafts to adjacent severed arteries in the host by well-defined and often tortuous segments termed junction vessels. The pre-operative vascular pattern was re-established when grafts were replaced without rotation but comparable survival of arteries also occurred when the grafted discs of skin were rotated. In most grafts, the segment of artery became stabilized by 4-9 weeks; more than 75% of the length of the arterial segments survived in 83% of grafts, with minimal accompanying histological changes in their walls. There was some increase in number and aborization of small vessels in the established grafts, although the vascular pattern was essentially similar to that of unoperated skin. The factors possibly involved in the survival of the large arteries in the grafts include the relative rigidity of the arterial wall, the early establishment of circulation, and the favourable haemodynamic position of the grafts.


Outcomes of systemic bivalirudin and sodium bicarbonate purge solution for Impella 5.5.
Impella 5.5 (Abiomed; Danvers, MA) (IMP5) is a commonly used, surgically implanted, tMCS device that requires systemic anticoagulation and purge solution to avoid pump failure. To avoid heparin-induced thrombocytopenia (HIT) from unfractionated heparin (UFH) use, our program has explored the utility of bivalirudin (BIV) for systemic anticoagulation and sodium bicarbonate-dextrose purge solution (SBPS) in IMP5.5. This single center, retrospective study included 34 patients supported on IMP5.5 with BIV based AC and SBPS between December 1st 2020 to December 1st 2021.The efficacy and safety end points were incidence of development of HIT, Tissue Plasminogen Activator (tPA) use for suspected pump thrombosis, stroke, and device failure as well as clinically significant bleeding. The median duration of IMP5.5 support was 9.8 days (IQR: 6-15). Most patients were bridged to HTX (58%) followed by recovery (27%) and LVAD implantation (15%). Patients were therapeutic on bivalirudin for 64% of their IMP5.5 support. One patient (2.9%) suffered from ischemic stroke and 26.5% (9) patients developed clinically significant bleeding. tPA was administered to 7(21%) patients. One patient in the entire cohort developed HIT. Our experience supports the use of systemic BIV and SBPS as a method to avoid heparin exposure in a patient population predisposed to the development of HIT.


[Radiodiagnosis of vesicoureteral reflux in the early postoperative period following adenectomy of the prostate gland].
Altogether 204 patients aged 43 to 81 were examined in the early postoperative period after adenomectomy of the prostate: 167 transvesical and 37 transureteral adenomectomies were performed. Vesicoureteral reflux (VUR) was usually noted on the 4th-5th day after operation therefore investigations aimed at its detection, were conducted at that time. Passive VUR was detected in 72 patients, active VUR in 31 patients. Both passive and active VUR were diagnosed in 23 patients. Passive VUR was detected by means of routine cystography, X-ray TV cystography, direct radionuclide cystography and ultrasonic investigation. The presence of VUR in ultrasonic investigation was established by the signs of dilatation of renal pelves and calyces in filling in of the bladder with antiseptic solution. Ultrasonic investigation made it possible to diagnose VUR of III-IV degree. Its sensitivity was 32%. The sensitivity of direct radionuclide cystography was 92%, that of cystography 51%. Active VUR was detected by miction cystography and X-ray TV miction cystography on the day of removal of drainage tubes from the bladder. The sensitivity of these two methods was 57 and 100%, respectively. Unlike the radionuclide and ultrasonic methods, these two methods made it possible to assess the state of the bladder cervix and urethra. The authors proposed an optimum sequence of the use of diagnostic methods for early detection of VUR and prevention of severe complications.


CBT for Generalized Anxiety Disorder in Parkinson's Disease: A Case Study.
To examine the use of cognitive behavioral therapy (CBT) in a case of co-occurring generalized anxiety disorder (GAD) and Parkinson's disease (PD). This case study refers to a male aged 75 years with a diagnosis of Idiopathic Parkinson's disease. It focuses on applying a CBT model to address the psychological difficulties with PD and GAD. This case study reveals key aspects in presentation, diagnosis, and psychological treatment between PD and GAD, and is one of few studies published in this area. Symptoms of anxiety in an older adult with PD decreased during a course of CBT. The implications of the treatment outcome of this study and further considerations of treatment plans for comorbid PD and anxiety have been discussed. Using CBT could positively impact non-motor symptoms of Parkinson's, such as sleep difficulties and speech impediments. Using CBT for the catastrophic thinking and worry content in GAD seems to act as a complementary therapy for psychological/non-motor symptoms of PD.


The Role of Tumor Markers: Carcinoembryonic Antigen and Cancer Antigen 15-3 in Patients With Breast Cancer.
Introduction Breast cancer is a major cause of mortality among females, worldwide. The present study was intended to evaluate the significance in the management of carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA15-3) in patients with breast cancer. Methodology A cohort study was conducted at the Jinnah Postgraduate Medical Center, Karachi, Pakistan from June 2020 to May 2021. All diagnosed cases of breast cancer who underwent surgical excision of tumor were eligible to partake. Patients who had metastatic breast cancer or had a recurrence were excluded. The patient's sociodemographic and clinical data were documented in a predefined pro forma. It included information about the age, sex, weight, as well as serum CEA and CA15-3. The CA15-3 and CEA levels for each patient were assessed by taking a 5ml blood sample and sending it to the laboratory for further workup. preoperatively on the second, seventh, and 28th postoperative days. Results A mean ± SD age of 52.6 ± 8.89 years was reported. Family history of breast cancer was positive in one-fourth of the patients. Nodal metastasis was negative in 114 (46.72%) patients. Three-fourth of patients had Stage II-IV with only a minority diagnosed with Stage I. The mean levels for CA15-3 in women with Stage I cancer was significantly lower on the seventh day and 28th postoperative day, compared to preoperative levels (p = 0.05). Similar associations were seen for stages II and III. Higher CEA levels were significantly associated with stage III breast cancer preoperatively (5.88 ng/ml, p = 0.05) compared to postoperative values. Conclusion The current study revealed that preoperative values of serum CEA and CA15-3 significantly reduced postoperatively. Moreover, patients with advanced cancers had significantly higher levels of both tumor markers than those with less advanced diseases. The current study highlighted the importance of regular assessment of serum CEA and CA15-3 in breast cancer patients. Both these biomarkers are substantially elevated in breast cancer patients, preoperatively. Determining the levels of serum CEA and CA15-3 pre- and postoperatively may determine the prognosis and aid in forming the most optimal patient care regime with respect to the stage and subtype of cancer.


Direct observation of Brownian dynamics of hard colloidal nanorods.
We synthesize monodisperse selenium (Se) colloidal rods, and the suspensions exhibit the smectic phase at a particle volume fraction (phi) of 0.28. Side-by-side rod clustering occurs at phi > 0.04. Cluster-size distributions and persistence times are determined for various phi. In dense suspensions (phi > 0.1), individual rods reveal characteristic fundamental motions, e.g., reptation and synchronized rotation. Mean-square displacements of the rods suggest a cage trapping and escape. Estimated translational and rotational diffusion coefficients show a large difference from predictions by computer simulations.


Genotyping-by-Sequencing Facilitates a High-Density Consensus Linkage Map for Aegilops umbellulata, a Wild Relative of Cultivated Wheat.
High-density genetic maps are useful to precisely localize QTL or genes that might be used to improve traits of nutritional and/or economical importance in crops. However, high-density genetic maps are lacking for most wild relatives of crop species, including wheat. Aegilops umbellulata is a wild relative of wheat known for its potential as a source of biotic and abiotic stress resistance genes. In this work, we have developed a framework consensus genetic map using two biparental populations derived from accessions PI 298905, PI 542369, PI 5422375, and PI 554395. The framework map comprised 3009 genotype-by-sequence SNPs with a total map size of 948.72 cM. On average, there were three SNPs per centimorgan for each chromosome. Chromosome 1U was the shortest (66.5 cM), with only 81 SNPs, whereas the remaining chromosomes had between 391 and 591 SNP markers. A total of 2395 unmapped SNPs were added to the linkage maps through a recombination frequency approach, and increased the number of SNPs placed on the consensus map to a total of 5404 markers. Segregation distortion was disproportionally high for chromosome 1U for both populations used to construct component linkage maps, and thus segregation distortion could be one of the probable reasons for the exceptionally reduced linkage size for chromosome 1U. From comparative analysis, Aeumbellulata chromosomes except 4U showed moderate to strong collinearity with corresponding homeologous chromosomes of hexaploid wheat and barley. The present consensus map may serve as a reference map in QTL mapping and validation projects, and also in genome assembly to develop a reference genome sequence for Ae. umbellulata.


Noninvasive bilevel positive pressure ventilation in patients with blunt thoracic trauma.
Noninvasive bilevel positive pressure ventilation (N-BiPAP) has an established role in providing respiratory support in patients with acute respiratory failure. The significant advantage of N-BiPAP is to avoid endotracheal intubation and its complications. Currently there are no data that support N-BiPAP as first-line treatment in patients with blunt thoracic trauma. To evaluate the safety and efficacy of N-BiPAP in patients with acute respiratory failure due to blunt thoracic trauma. Prospective observational study. Twenty-two patients with blunt chest trauma (mean injury severity score 26 +/- 9) were studied. N-BiPAP was applied via a tight-fitting full or total-face mask, combined with regional anesthesia in all patients. N-BiPAP resulted in significant changes in blood gasses, heart rate and breathing frequency at 1 h. Eighteen out of 22 patients avoided intubation and were discharged from the ICU (success group). Four patients met predefined criteria and required intubation (failure group) within 24 h after N-BiPAP. Three of the patients in the failure group survived while 1 developed septic shock and died. The acute response of oxygenation to N-BiPAP differed significantly between groups, being higher in the success group. Complications related to N-BiPAP were minor, consisting of nose bridge injury (1 patient) and gastric distention (1 patient). N-BiPAP administration could be a safe and effective method to improve the gas exchange in patients with acute respiratory failure due to blunt thoracic trauma.


Long-term effect of stenting after an endoscopic modified Lothrop procedure.
The aim of this study was to study the effect of stenting with a silastic sheet on the long-term patency of the common frontal ostium after an endoscopic modified Lothrop procedure (EMLP). A retrospective analysis was performed of prospectively collected data at an academic tertiary referral center, where 72 patients underwent an EMLP between 1996 and 2003. Patency of the common frontal ostium was studied as an ordinal variable with three classes (patent, stenotic, and closed). A common ostium was endoscopically visualized in 94% of the patients (61.1% patent and 33.3% stenotic). Symptoms improved in 75% of the patients. A significant positive correlation was found between ostium patency and symptom improvement. Twenty-five patients were stented postoperatively. There was no statistical difference between the stented and the nonstented groups with regards to ostium patency and symptoms improvement. Short-term stenting does not appear to reduce the rate of postoperative stenosis of the common frontal ostium after an EMLP.


Neutrophilia and NETopathy as Key Pathologic Drivers of Progressive Lung Impairment in Patients With COVID-19.
There is an urgent need for new therapeutic strategies to contain the spread of the novel coronavirus disease 2019 (COVID-19) and to curtail its most severe complications. Severely ill patients experience pathologic manifestations of acute respiratory distress syndrome (ARDS), and clinical reports demonstrate striking neutrophilia, elevated levels of multiple cytokines, and an exaggerated inflammatory response in fatal COVID-19. Mechanical respirator devices are the most widely applied therapy for ARDS in COVID-19, yet mechanical ventilation achieves strikingly poor survival. Many patients, who recover, experience impaired cognition or physical disability. In this review, we argue the need to develop therapies aimed at inhibiting neutrophil recruitment, activation, degranulation, and neutrophil extracellular trap (NET) release. Moreover, we suggest that currently available pharmacologic approaches should be tested as treatments for ARDS in COVID-19. In our view, targeting host-mediated immunopathology holds promise to alleviate progressive pathologic complications of ARDS and reduce morbidities and mortalities in severely ill patients with COVID-19.


[Strategies for efficacy and function positioning of new resources of Chinese materia medica].
Chinese materia medica( CMM) serves as an important cornerstone for the development of traditional Chinese medicine( TCM) culture and industry due to its unique ecological,medical,economic,scientific and technological,and cultural values. The supply shortage and unstable quality of some CMM resources have hindered the development of TCM. Ensuring the sustainable use of CMM resources has become essential for the development of TCM in China. Enriching CMM resources is the key to ensuring the sustainable utilization of TCM resources in China,which can be achieved via expanding the medicinal parts,developing the substitutes,seeking for analogues,exploring the ethnic and folk medicines,or introducing foreign medicinal materials. CMM efficacy or function positioning plays a very important role in the transformation of new CMM resources. The strategies and methods for efficacy or function positioning of new CMM resources,including analogy,plant genetic relationship exploration,medicinal property deduction,ethnobotanical investigation,text mining,network pharmacology,and structure-activity relationship exploration,were systematically proposed in this study based on CMM theory,textual research,and modern methodologies. This paper is expected to provide a theoretical reference for the continuous enrichment and development of CMM resources and the high-quality development of TCM culture and industry.


Practice Environment Scale of the Nursing Work Index: A Reliability Generalization Meta-Analysis.
A healthy work environment is a critical factor in nurse satisfaction, retention, and patient outcomes. The Practice Environment Scale of the Nursing Work Index (PES-NWI) is the most commonly used instrument to measure the nursing practice environment. This study uses meta-analysis to examine the reliability generalization of the PES-NWI. A meta-analysis of 51 studies representing a total of 80,563 subjects was conducted. The mean score reliability for the PES-NWI based on 38 studies (n = 68,278) was .922 (p < .05). The Mean Weighted Effect Size was stronger for studies conducted in the United States versus non-U.S. (.946 vs. .907). For studies rated high and moderate quality, the mean score reliability was .911 and .946, respectively. Scores on the PES-NWI are reliable for measuring the nursing practice environment across samples in the United States and non-U.S. countries.


The prolactin gene, but not the growth hormone gene, is located on chromosome 9 in the rat.
Experiments from a number of laboratories, employing clonal lines of rat pituitary tumor cells (GH cells), have shown that the cellular levels of the mRNAs for prolactin and growth hormone are subject to regulation by a variety of hormones and other factors. To provide a framework for further studies of the regulation of the expression of these two genes, we have studied their chromosomal locations in the rat, employing our recently developed technique for identifying a unique gene by direct hybridization in situ with a 125I-labeled cDNA plasmid. A rat prolactin cDNA plasmid ( pPRL -1) yielded a signal significantly above that expected on a random basis over chromosomes 9 and 15 from rat FRE cells. Both chromosomes exhibited a subchromosomal clustering of silver grains near the telomere of chromosome 9 and on the short arm of chromosome 15. Control experiments with plasmid pBR322 also yielded a significant signal clustered over the short arm of chromosome 15. Hence, we conclude that chromosome 15 contains sequences homologous to pBR322 and that the prolactin gene is located near the telomere of chromosome 9 in the rat. A rat growth hormone cDNA plasmid (pBR322-GH1) yielded a signal above that expected on a random basis over FRE cell chromosomes 7, 13, and 15, although the labeling was poorer than with prolactin. The signal over chromosome 15 is probably due to sequences homologous to plasmid pBR322. The signals over chromosomes 7 and 13 showed a clear subchromosomal clustering near the centromere on chromosome 7 and near the telomere on the long arm of chromosome 13.(ABSTRACT TRUNCATED AT 250 WORDS)


Regulation of terminal differentiation of cultured human keratinocytes by vitamin A.
Vitamin A is known to exert an important influence on epithelial differentiation. The fetal calf serum supplement of cell-culture medium contains enough of the vitamin to affect the differentiation of cultured keratinocytes derived from epidermis and from other stratified squamous epithelia. The cellular and molecular properties of the cultures are altered when the medium is supplemented with serum from which the vitamin A has been removed by solvent extraction (delipidized serum). Cell motility is reduced, the adhesiveness of cells increases and pattern formation is prevented. In both epidermal and conjunctival keratinocytes, removal of vitamin A leads to the synthesis of a 67 kd keratin characteristic of terminally differentiating epidermis and to much reduced synthesis of the 52 kd and 40 kd keratins typical of conjunctiva. These changes, both cellular and molecular, are reversed by the addition of retinyl acetate to the medium containing delipidized serum. Cell motility and pattern formation are restored, and detachment of the most mature cells from the surface of the stratified epithelium is promoted. Synthesis of the 67 kd keratin is prevented and the synthesis of the 40 and 52 kd keratins is stimulated. The nature of the keratins synthesized is regulated by the concentration of vitamin A, and each cell type adjusts its synthesis differently at a given vitamin concentration.


Considerations when calculating the sample size for an inequality test.
Click here for Korean Translation. Calculating the sample size is a vital step during the planning of a study in order to ensure the desired power for detecting clinically meaningful differences. However, estimating the sample size is not always straightforward. A number of key components should be considered to calculate a suitable sample size. In this paper, general considerations for conducting sample size calculations for inequality tests are summarized.


Technological Development and Application of Plant Genetic Transformation.
Genetic transformation is an important strategy for enhancing plant biomass or resistance in response to adverse environments and population growth by imparting desirable genetic characteristics. Research on plant genetic transformation technology can promote the functional analysis of plant genes, the utilization of excellent traits, and precise breeding. Various technologies of genetic transformation have been continuously discovered and developed for convenient manipulation and high efficiency, mainly involving the delivery of exogenous genes and regeneration of transformed plants. Here, currently developed genetic transformation technologies were expounded and compared. Agrobacterium-mediated gene delivery methods are commonly used as direct genetic transformation, as well as external force-mediated ways such as particle bombardment, electroporation, silicon carbide whiskers, and pollen tubes as indirect ones. The regeneration of transformed plants usually involves the de novo organogenesis or somatic embryogenesis pathway of the explants. Ectopic expression of morphogenetic transcription factors (Bbm, Wus2, and GRF-GIF) can significantly improve plant regeneration efficiency and enable the transformation of some hard-to-transform plant genotypes. Meanwhile, some limitations in these gene transfer methods were compared including genotype dependence, low transformation efficiency, and plant tissue damage, and recently developed flexible approaches for plant genotype transformation are discussed regarding how gene delivery and regeneration strategies can be optimized to overcome species and genotype dependence. This review summarizes the principles of various techniques for plant genetic transformation and discusses their application scope and limiting factors, which can provide a reference for plant transgenic breeding.


New Methods for Replacing Single Missing Teeth with Non-prep Bridges (NPBs) - A Case Series.
Newly developed non-invasive methods for replace a missing tooth and closing single-tooth gaps in the poster- ior region using resin composite are presented. Four different non-invasive methods and the technical procedures, materials and instruments used are presented in a case series. These include the direct intraoral insertion of composite (with and without individual shaping aids) and indirect restorations, which are fabricated conventionally or digitally and bonded. The case series showed that all four methods can be used to replace single missing teeth in the posterior region, meeting current clinical requirements. Particular attention was paid to the design of the pontics, the dimension of the connector area, firm proximal contacts to the adjacent teeth, hygiene, and appearance of the non-prep bridges (NPBs). The advantages and disadvantages for both the direct and indirect techniques illustrated in this case series were com- pared in detail. Several direct and indirect non-invasive methods for single-tooth replacement are available today. Although the evidence is still limited, there is a potential for frugal dental interventions with NPBs. Further experimental and clinical studies are necessary to demonstrate that they reliably meet quality requirements (including sufficient survival rates), satisfy the criteria of cost-effectiveness (compared to treatment alternatives) and that there is a demand from the population.


Recurrent pleomorphic adenoma of the parotid.
Nineteen patients with recurrent pleomorphic adenoma of the parotid are reported. The aetiology of recurrence is discussed and capsule rupture is emphasized as a major cause, especially of multinodular recurrence. Further surgical attempts to remove recurrences are justified by the possibility of misdiagnosis of the original tumour and the late development of malignancy in recurrent nodules. Tumour clearance was achieved in all but two of 19 patients with recurrence or incomplete primary excision in those in whom it was attempted. Radiotherapy does not seem to be an appropriate alternative to repeated surgical excision.


The role of the preserved sequences of Dam methylase.
We have undertaken a site directed mutational analysis of two of the preserved regions in the amino acid sequence of Dam methylase in order to characterize their role. Mutations in region IV (sequence DPPY) abolish catalytic activity and greatly affect AdoMet crosslinking. Mutants in region III display a lowered specific activity with an unchanged AdoMet crosslinking capacity. We have also made a series of deletions both at the N and C terminal parts of the protein, which have been found to provide inactive enzyme. We discuss the significance of these results for the understanding of the functional properties of the enzyme.


The effect of dimethylphenylpiperazinium on the amine uptake process.
Dimethylphenylpiperazinium (DMPP) potentiated the contractile response of rat isolated vas deferens to norepineprine and decreased that to tyramine. This action of DMPP is explained by the observation that it inhibited the uptake of 3H-norepinephrine and 3H-tyramine into adrenergic nerve terminals.


Interconversion studies of betamethasone acetate polymorphs.
The polymorph interconversions of Betamethasone Acetate (BA) were studied under various pharmaceutical conditions, such as grinding, heating and suspending in water, based on differential scanning calorimetry, thermogravimetric analysis, and X-ray powder diffraction. There existed enantiotropic relationships between the three polymorphs of BA, which were named form II, Ialpha, and Ibeta work, respectively. It was concluded that form II was the most stable form when suspended in water.


Early recognition of dementia by nurses.
Early recognition of dementia is a key policy objective designed to maximize the efficacy of treatment and to provide timely and appropriate support before crisis occurs. The impact of early recognition of dementia is under-researched in primary care nursing. To explore whether community mental health nurses (CMHNs), community nurses (CNs) and practice nurses (PNs) have different perspectives on early diagnosis of dementia and to consider the possible effects of any variation. Data are drawn from questionnaires completed by CMHNs (79), CNs (153) and PNs (36) who attended workshops offered on 24 occasions in 21 settings across the United Kingdom. The workshops attracted a range of primary care practitioners and were part of a training programme on early diagnosis of dementia. Analysis of the data shows some differences in knowledge, experience and confidence between the three nurse groups. CMHNs were more confident in their abilities to recognize dementia and found providing support less difficult than CNs and PNs. CMHNs considered that they were best placed to co-ordinate services for people with newly recognized dementia. CNs and PNs, however, reported experience of working with people with dementia and many appeared able to respond to early signs and to identify potential sources of support. While CMHNs may have a key role in responding effectively to the newly identified needs of people with early dementia, other nurses working in the community are likely to encounter people with early dementia. In the context of a policy objective to identify people with dementia earlier, all community-based nurses should be able to recognize the possibility of dementia and support those undergoing referral or assessment. Their confidence in doing so should be enhanced by continued professional development. Training in dementia recognition, involvement in and membership of primary care teams supporting people with dementia should not be confined to CMHNs. Nurses who regularly encounter the general population of older people may be well placed to provide continuity of support for those who may, or may not, have cause to suspect that they or their relatives have early dementia.


Current knowledge and treatment strategies for grade II gliomas.
World Health Organization grade II gliomas (GIIGs) include diffuse astrocytoma, oligodendroglioma, and oligoastrocytoma. GIIG is a malignant brain tumor for which the treatment outcome can still be improved. Review of previous clinical trials found the following: (1) GIIG increased in size by 3-5 mm per year when observed or treated with surgery alone; (2) after pathological diagnosis, the survival rate was increased by early aggressive tumor removal at an earlier stage compared to observation alone; (3) although the prognosis after total tumor removal was significantly better than that after partial tumor removal, half of the patients relapsed within 5 years; (4) comparing postoperative early radiotherapy (RT) and non-early RT after relapse, early RT prolonged progression-free survival (PFS) but did not affect overall survival (OS); (5) local RT of 45 to 64.8 Gy did not impact PFS or OS; (6) in patients with residual tumors, RT combined with chemotherapy (procarbazine plus lomustine plus vincristine) prolonged PFS compared with RT alone but did not affect OS; and (7) poor prognostic factors included astrocytoma, non-total tumor removal, age ≥40 years, largest tumor diameter ≥4-6 cm, tumor crossing the midline, and neurological deficit. To improve treatment outcomes, surgery with functional brain mapping or intraoperative magnetic resonance imaging or chemoradiotherapy with temozolomide is important. In this review, current knowledge regarding GIIG is described and treatment strategies are explored.


Relationships Between Lifestyle, Quality of Life, Organizational Culture and Job Satisfaction Among Nurses Related to Health Promoting Hospital (HPH) Standards.
Providing quality healthcare services through health promotion activities to patients, hospital-based professionals and the wider community is the goal of the health promoting hospital (HPH). There is, however, no formal structured pathway for "universally" providing health promotion services in hospitals. Accordingly, this study was conducted with the aim of presenting a model designed to promote and increase health-related satisfaction of hospital-professionals in health-promoting hospitals (HPHs) in Iran-as a potential tool to guide international HPH standards. Lifestyle, quality of life, organizational culture, and job satisfaction were measured using standardized questionnaires in specialized hospitals in Hamadan, Iran. A structural equation model (SEM) using partial least squares (PLS) software (version 2) was used to determine the validity and fit of the conceptual framework/model. The study revealed that several factors were identified as strong predictors of job satisfaction and wellbeing, including various dimensions of lifestyle such as spiritual health, physical activity, stress management, and interpersonal communication, dimensions of quality of life including physical and mental aspects, and organizational culture. The values of predictive relevance (Q2) for physical and psychological dimension of life quality, organizational culture, and job satisfaction were estimated to be 0.101, 0.250, and 0.040 and 0.251, respectively. Conclusively, the study found a goodness of fit (GOF) value of 0.415, indicating that the model had a high predictive power and fit well. Based on these results, it is suggested that implementing HPH interventions that focus on the outcomes of this model could lead to increased job satisfaction and wellbeing in hospitals. Additionally, the model could serve as a useful indicator of HPHs.


Structural basis of the XPB-Bax1 complex as a dynamic helicase-nuclease machinery for DNA repair.
Nucleotide excision repair (NER) is a major DNA repair pathway for a variety of DNA lesions. XPB plays a key role in DNA opening at damage sites and coordinating damage incision by nucleases. XPB is conserved from archaea to human. In archaea, XPB is associated with a nuclease Bax1. Here we report crystal structures of XPB in complex with Bax1 from Archaeoglobus fulgidus (Af) and Sulfolobus tokodaii (St). These structures reveal for the first time four domains in Bax1, which interacts with XPB mainly through its N-terminal domain. A Cas2-like domain likely helps to position Bax1 at the forked DNA allowing the nuclease domain to incise one arm of the fork. Bax1 exists in monomer or homodimer but forms a heterodimer exclusively with XPB. StBax1 keeps StXPB in a closed conformation and stimulates ATP hydrolysis by XPB while AfBax1 maintains AfXPB in the open conformation and reduces its ATPase activity. Bax1 contains two distinguished nuclease active sites to presumably incise DNA damage. Our results demonstrate that protein-protein interactions regulate the activities of XPB ATPase and Bax1 nuclease. These structures provide a platform to understand the XPB-nuclease interactions important for the coordination of DNA unwinding and damage incision in eukaryotic NER.


Rapid microscope auto-focus method for uneven surfaces based on image fusion.
Microscopic imaging of uneven surfaces is difficult because of the limited depth of field. In this study, we developed a rapid auto-focus method for uneven surfaces based on image fusion. The Prewitt operator was used to detect the vertical edges of the images. Then, the focus position was theoretically calculated using a Gaussian function, and image fusion was applied to obtain the final in-focus image. An experiment was designed to verify the developed method. The results revealed that this method is effective for printed circuit boards.


Precision Medicine and/or Biomarker Based Therapy in T2DM: Ready for Prime Time?
Approximately 30-40% of people with type 2 diabetes mellitus develop chronic kidney disease. This is characterised by elevated blood pressure, declining kidney function and enhanced cardiovascular morbidity and mortality. Increased albuminuria and decreasing estimated glomerular function has to be evaluated regularly to diagsnose kidney disease. New biomarkers may facilitate early diagnosis and provide infomation on undlying pathology thereby supporting early precision intervention for the optimal benefit. A number of biomarkers have been suggested but are not yet implemented in clinical practice. iI the future such bimarkers may pave the way for personalized treatment.


Duplication and triplication with staggered breakpoints in human mitochondrial DNA.
We identified a tandem duplication and triplication of a mitochondrial DNA (mtDNA) segment in the muscle of a 57-year-old man with no evidence of a neuromuscular disorder. A large triplication of a mtDNA coding region has not been previously reported in humans. Furthermore, the rearrangements (comprising 10-12% of the muscle mtDNA pool in the propositus) were unique because the breakpoints were staggered at both ends (between mtDNA positions 3263-3272 and 16,065-16,076) and contained no identifiable direct repeats. Both sides of the breakpoint were located approximately 35 bp downstream of regions that undergo frequent strand displacement by either transcription (positions 3263-3272) or replication (positions 16,065-16,076), suggesting that topological changes generated by the movement of RNA/DNA polymerases may be associated with the genesis of a subclass of mtDNA rearrangements. The presence of low levels of these rearrangements in other normal adults also suggest that these mutations are not rare. The characterization of these rearrangements shed light on potential alternative mechanisms for the genesis of mtDNA rearrangements.


[A case of arteriovenous malformation seen as an endobronchial lesion].
Pulmonary arteriovenous malformation (PAVM) is an abnormal communication between pulmonary arteries and pulmonary veins, and congenital form is seen more prevalently. The classic radiological appearance is a round, well-circumscribed lesion. PAVM was observed in a 22-year-old male as an endobronchial lesion, and treated by wedge resection. Reviewing the literature, we identified only one case report that describes endobronchial view of PAVM.


E-Cigarette Use Among Adolescents Not Susceptible to Using Cigarettes.
Research suggests that adolescents who use electronic cigarettes (e-cigarettes), including adolescents not susceptible to smoking cigarettes (ie, those who have never smoked cigarettes and are not attitudinally susceptible to using cigarettes), are more likely to initiate using cigarettes or other combustible tobacco products than adolescents who do not use e-cigarettes. In this study, we examined correlates of e-cigarette use and susceptibility among adolescents not susceptible to future cigarette smoking. We used data on high school students from the 2015 North Carolina Youth Tobacco Survey (n = 1,627). SAS logistic regression survey procedures were used to account for the complex survey design and sampling weights. Increasing perceived harm of e-cigarettes was associated with lower odds of susceptibility to using e-cigarettes (adjusted odds ratio [AOR] = 0.79; 95% confidence interval [CI], 0.65-0.96) and current use of e-cigarettes (AOR = 0.43; 95% CI, 0.25-0.72). Similar patterns were found for perceived harm of secondhand e-cigarette vapor. Exposure to e-cigarette vapor in indoor or outdoor public places was positively associated with susceptibility to using e-cigarettes (AOR = 1.96; 95% CI, 1.33-2.91) and with current e-cigarette use (AOR = 5.69; 95% CI, 2.57-12.61). To prevent initiation of e-cigarette use, particularly among adolescents not susceptible to smoking cigarettes, educational campaigns could target harm perceptions associated with e-cigarettes. In addition, regulations that limit adolescents' exposure to e-cigarettes in public places may decrease e-cigarette use by nonsusceptible adolescents.


Evaluation of in vivo cytogenetic toxicity of europium hydroxide nanorods (EHNs) in male and female Swiss albino mice.
Our group already demonstrated that europium hydroxide nanorods (EHNs) show none or mild toxicity in C57BL/6 mice even at high dose and exhibited excellent pro-angiogenic activity towards in vitro and in vivo models. In the present study, we evaluated the in vivo cytogenetic toxicity of intraperitoneally administered EHNs (12.5-250 mg/kg/b.w.) in male and female Swiss albino mice by analyzing chromosomal aberrations (CAs), mitotic index (MI), micronucleus (MN) from bone marrow and peripheral blood. Furthermore, we performed the cytogenetic toxicity study of EHNs towards Chinese hamster ovary (CHO) cells, in order to compare with the in vivo results. The results of CA assay of mice treated with EHNs (12.5-125 mg/kg/b.w.) showed no significant change in the formation of aberrant metaphases compared to the control group. Also, there was no significant difference in the number of dividing cells between the control group and EHNs-treated groups observed by MI study, suggesting the non-cytotoxicity of EHNs. Additionally, FACS study revealed that EHNs do not arrest cells at any phase of cell cycle in the mouse model. Furthermore, MN test of both bone marrow and peripheral blood showed no significant differences in the induction of MNs when compared with the control group. In vitro results from CHO cells also support our in vivo observations. Considering the role of angiogenesis by EHNs and the absence of its genotoxicity in mouse model, we strongly believe the future application of EHNs in treating various diseases, where angiogenesis plays an important role such as cardiovascular diseases, ischemic diseases and wound healing.


Ascomycete fungal actins differentially support plant spatial cell and organ development.
Actin interacts with a wide variety of cytoplasmic and nuclear proteins to support spatial development in nearly all eukaryotes. Null mutations in plant vegetative actins produce dramatically altered cell, tissue, and organ morphologies. Animal cytoplasmic actins (e.g., human HsACTB, HsACTG1) and some ancestral protist actins fully suppress these mutant phenotypes suggesting that some animal, plant, and protist actins share functional competence for spatial development. Considering that fungi have a phylogenetic origin closer to animals than plants, we were interested to explore whether the fungal actins may have this same capacity to function in plants and support development. We ectopically expressed actins from four highly divergent ascomycete fungi in two different Arabidopsis double vegetative actin null mutants. We found that expression of actin from the earliest diverging ascomycete subphyla, the archiascomycete Schizosaccharomyces pombe, qualitatively and quantitatively suppressed the root cell polarity and root organ developmental defects of act8/act7 mutants and the root-hairless cell elongation phenotype of act2/act8 mutants. Interestingly, the actin from the pyrenomycete Neurospora crassa was modestly effective in the suppression of vegetative actin mutant phenotypes. In contrast, actins from the saccharomycetes Saccharomyces cerevisiae and Candida albicans were unable to support any aspect of plant development, and moreover induced severe dwarfism and sterility. These data imply that basal fungi inherited an actin with full competence for spatial development from their protist ancestor and maintained it via non-progressive sequence evolution, while the later more derived fungal species lost these activities.


Sulfonyl hydrazine-functionalized polymer as a specific capturer of reducing glycans from complex samples for high-throughput analysis by electrospray ionization mass spectrometry.
Qualitative and quantitative studies of glycosylation patterns of various biologically important proteins represent a key field for the understanding of their complex structure-function relationships. However, the analysis of glycoprotein glycans is usually undermined by tedious sample processing steps prior to detection, including deproteination, desalting and removal of some other non-glycan impurities, which results in considerable sample loss and increased difficulty of quantitative analysis. Herein we report a facile and versatile method for the quantitative isolation of reducing glycans from complex samples using sulfonyl hydrazine-functionalized polystyrene (SHPS) beads, namely the SHPS-based glycan capturing procedure. This method allows the chemoselective and efficient condensation of the aldehyde group of reducing glycans with the active sulfonyl hydrazine group of SHPS beads under anhydrous conditions, resulting in the formation of sulfonyl hydrazone conjugates. The non-glycan components in samples, such as proteins, salts and some other impurities, can be completely removed by washing the sulfonyl hydrazone conjugates. Regeneration of the reducing glycans can be performed via mild hydrolysis of the washed sulfonyl hydrazone conjugates. This procedure is compatible with almost all the current techniques for the derivatization or detection of reducing glycans. We have obtained essential data for this method, including optimized reaction conditions, linearity and reproducibility for glycan quantitation, as well as a final glycan recovery ratio. Moreover, mass spectrometric analysis of the glycans from some complex biological samples, including milk, human blood plasma and fetal bovine serum, was achieved, demonstrating good applicability of this novel procedure.


Integrating Clinical Cancer and PTM Proteomics Data Identifies a Mechanism of ACK1 Kinase Activation.
Beyond the most common oncogenes activated by mutation (mut-drivers) there likely exists a variety of low-frequency mut-drivers, each of which is a possible frontier for targeted therapy. To identify new and understudied mut-drivers, we developed a machine learning (ML) model that integrates curated clinical cancer data and post-translational modification (PTM) proteomics databases. We applied the approach to 62,746 patient cancers spanning 84 cancer types and predicted 3,964 oncogenic mutations across 1,148 genes, many of which disrupt PTMs of known and unknown function. The list of putative mut-drivers includes established drivers and others with poorly understood roles in cancer. This ML model is available as a web application. As a case study, we focused the approach on non-receptor tyrosine kinases (NRTKs) and found a recurrent mutation in activated CDC42 kinase-1 (ACK1) that disrupts the Mig6 homology region (MHR) and ubiquitin-association (UBA) domains on the ACK1 C-terminus. By studying these domains in cultured cells, we found that disruption of the MHR domain helps activate the kinase while disruption of the UBA increases kinase stability by blocking its lysosomal degradation. This ACK1 mutation is analogous to lymphoma-associated mutations in its sister kinase, TNK1, which also disrupt a C-terminal inhibitory motif and UBA domain. This study establishes a mut-driver discovery tool for the research community and identifies a mechanism of ACK1 hyperactivation shared among ACK family kinases. Implications: This research identifies a potentially targetable activating mutation in ACK1 and other possible oncogenic mutations, including PTM-disrupting mutations, for further study.


Ambient tonicity and intestinal cytochrome CYP3A.
Recently-discovered tonicity-dependence of human CYP3A expression in vitro may be a novel mechanism of CYP3A regulation in the intestinal epithelia, which exists in a dynamic osmotic environment influenced by food intake. A combination of focused and comprehensive literature searches to identify any relevant reports using Medline (from 1950 to 7 November 2009) through the OVID system. An update on current knowledge on osmotic environment in the gastrointestinal (GI) tract and its impact on intestinal CYP3A expression and function with special emphasis on the tonicity-sensitive transcription factor nuclear factor of activated T cells 5 (NFAT5). In vitro hypertonicity of ambient osmotic environment in cultured human cells increases expression of CYP3A through transcriptional enhancement by osmosensitive NFAT5. Although post-prandial osmolality in the GI lumen in vivo is substantially increased, NFAT5 activation has not been reported. Similarly, high-salt diet increases intestinal CYP3A function in humans, but it is not known whether these changes are mediated directly by NFAT5.


Obesity and hypertension: epidemiology, mechanisms, treatment.
There is a close epidemiological association between obesity and elevated blood pressure for all age groups, although not every obese individual becomes hypertensive. In populations without age-related increases in body weight, an elevation of blood pressure with age is not seen. Mechanisms included in the development of hypertension in obesity are hyperinsulinemia, insulin induced sodium retention and increased sympathetic tone. Overnutrition with over intake of sodium and lack of physical exercise contribute to the metabolic syndrome of obesity. Thus, weight reduction by decreased energy uptake and increased physical exercise is recommended in the treatment of hypertension in obese patients. The resulting fall in insulin levels may lead to decreased sodium absorption in the kidney. Although treatment of obesity by weight loss decreases blood pressure substantially, a minority of patients do not respond to the weight loss. Blood pressure generally decreases before normal weight is achieved. Salt intake reduction does not appear to explain why weight reduction lowers blood pressure. Reduced levels of plasma renin activity, serum aldosterone levels, catecholamine levels and serum insulin levels may be involved in the blood pressure lowering associated with weight loss. Since the risk of cardiovascular disease in the hypertensive patient is not only determined by the blood pressure, an overall treatment which aims at reduction of other risk factors such as glucose intolerance and hyperlipoproteinemia is advocated. Thus, in any obese hypertensive patient normalization of excess body weight and increased physical activity appears to be the first and most important step of any rational therapeutic strategy.


Higher order, multifeatural object encoding by the oculomotor system.
Previous behavioral and physiological research has demonstrated that as the behavioral relevance of potential saccade goals increases, they elicit more competition during target selection processing as evidenced by increased saccade curvature and neural activity. However, these effects have only been demonstrated for lower order feature singletons, and it remains unclear whether more complicated featural differences between higher order objects also elicit vector modulation. Therefore, we measured human saccades curvature elicited by distractors bilaterally flanking a target during a visual search saccade task and systematically varied subsets of features shared between the two distractors and the target, referred to as objective similarity (OS). Our results demonstrate that saccades deviated away from the distractor highest in OS to the target and that there was a linear relationship between the magnitude of saccade deviation and the number of feature differences between the most similar distractor and the target. Furthermore, an analysis of curvature over the time course of the saccade demonstrated that curvature only occurred in the first 20-30 ms of the movement. Given the multifeatural complexity of the novel stimuli, these results suggest that saccadic target selection processing involves dynamically reweighting vector representations for movement planning to several possible targets based on their behavioral relevance. NEW & NOTEWORTHY We demonstrate that small featural differences between unfamiliar, higher order object representations modulate vector weights during saccadic target selection processing. Such effects have previously only been demonstrated for familiar, simple feature singletons (e.g., color) in which features characterize entire objects. The complexity and novelty of our stimuli suggest that the oculomotor system dynamically receives visual/cognitive information processed in the higher order representational networks of the cortical visual processing hierarchy and integrates this information for saccadic movement planning.


Significance tests for covariates in the diagnostic accuracy index of a biomarker against a continuous gold standard.
Receiver operating characteristic (ROC) curve is a popular tool to describe and compare the diagnostic accuracy of biomarkers when a binary-scale gold standard is available. However, there are many examples of diagnostic tests whose gold standards are continuous. Hence, Several extensions of receiver operating characteristic (ROC) curve are proposed to evaluate the diagnostic potential of biomarkers when the gold standard is continuous-scale. Moreover, in evaluating these biomarkers, it is often necessary to consider the effects of covariates on the diagnostic accuracy of the biomarker of interest. Covariates may include subject characteristics, expertise of the test operator, test procedures or aspects of specimen handling. Applying the covariate adjustment to the case that the gold standard is continuous is challenging and has not been addressed in the literature. To fill the gap, we propose two general testing frameworks to account for the covariates effect on diagnostic accuracy. Simulation studies are conducted to compare the proposed tests. Data from a study that assessed three types of imaging modalities with the purpose of detecting neoplastic colon polyps and cancers are used to illustrate the proposed methods.


COVID-19 anxiety and mental health among university students during the early phases of the U.S. pandemic.
This study examined the impact of COVID-19 in the early stages of the pandemic on university students in the U.S. by: (1) characterizing COVID-19-related disruptions; (2) evaluating health anxiety, obsessive-compulsive (OC), depression, and stress symptoms; and (3) analyzing the unique role of COVID-19 anxiety on mental health outcomes, after accounting for relevant variables. Participants included 263 students (63.9% female). Data were collected online between March 19, 2020 and May 1, 2020. Participants screened positive for health anxiety (6.5%), OC symptoms (48.7%), or depression (29.7%). COVID-19 anxiety was positively associated with mental health symptoms. After controlling for demographics and COVID-19 impact, COVID-19 anxiety accounted for significant variance in health anxiety, OC symptoms, and stress. Findings demonstrate the vast impact of COVID-19 on mental health among university students and provide guidance for identifying mental health priorities in the context of public health crises.


Studies on the mode of action of tomatine as a fungitoxic agent.
Removal of 1 or more sugar residues from the alpha-tomatine molecule markedly decreased its fungitoxicity. While partial hydrolysis of alpha-tomatine did not greatly affect its surfactant properties, it did destroy the ability of this alkaloid to form a complex with cholesterol. Only unprotonated alpha-tomatine was capable of binding cholesterol; the protonated form did not. Since alpha-tomatine was far more toxic at a high pH than at a low pH, this suggests that the unprotonated alkaloid is the active form and that it acts by complexing with fungal sterols.


The excision mechanism in reverse transcriptase: pyrophosphate leaving and fingers opening are uncoupled events with the analogues AZT and d4T.
We conducted molecular dynamics simulations of complexes of HIV-1 reverse transcriptase (RT) with the substrate and the antiretrovirals AZT and d4T for which resistance emerges via the excision mechanism. It is currently believed that excision results from the inability of AZT to translocate to the P site because of the steric hindrances imposed by the azide group. However, such explanation is far from satisfactory as d4T does not have such steric hindrances and still suffers from excision. Such contradiction motivated us for the present study. The results point to a new explanation for excision. RT preferably excises these inhibitors over the substrate as a consequence of a different pattern of hydrogen bridges they establish with the N site after incorporation. In the complexes with normal nucleotides, the fingers residues K65 and R72 establish hydrogen bonds mainly with the leaving PPi. With the inhibitors, those same residues establish hydrogen bonds primarily with the substituted nucleotides. Consequently, pyrophosphate is eliminated before the opening of the fingers domain, which allows ATP binding, with subsequent excision and development of drug resistance.


Lithium toxicity: an iatrogenic problem in susceptible individuals.
Lithium toxicity, manifesting primarily as neurotoxicity, is a significant health problem and is primarily iatrogenic in nature. Despite 50 years of medical experience with lithium, factors contributing to the development of severe neurotoxicity remain poorly documented. We hypothesized that severe neurotoxicity represents the most clinically significant manifestation of lithium toxicity. We proposed that this occurs primarily in the context of chronic therapeutic administration ('chronic poisoning'), rather than in the context of an overdose. Furthermore we hypothesized that patients who developed chronic poisoning did so in the presence of identifiable factors which predictably impair lithium clearance. A retrospective analysis of 97 cases of lithium poisoning, treated at a regional centre over a 13-year period was performed. Demographic data and factors considered likely to relate to the risk of developing lithium toxicity were recorded. Patients were classified according to mode of poisoning (acute, acute on chronic, or chronic) and according to severity of neurotoxicity (nil, mild, moderate, severe). The risk of developing severe neurotoxicity as a result of each mode of poisoning was assessed. The association between various risk factors and the development of chronic poisoning was assessed using a logistic regression model. Twenty-eight cases were rated as suffering severe neurotoxicity; in 26 this developed in the context of chronic poisoning and in two in the context of acute on chronic poisoning. All patients who developed severe neurotoxicity had at least one putative risk factor present, regardless of mode of poisoning. Length of stay was significantly longer for cases with severe neurotoxicity compared to those without severe neurotoxicity (12 vs. 2 days, P < 0.001). Peak serum lithium concentrations were significantly higher in cases with severe neurotoxicity compared to those without (2.3 vs. 1.6 mmol/L, P = 0.02). Patients presenting with chronic poisoning had a substantially higher risk of severe neurotoxicity than those presenting after an overdose of lithium (Odds Ratio [OR] 136, 95% CI 23-1300). A logistic regression model showed three factors contributed independently to the risk of chronic poisoning. These were: nephrogenic diabetes insipidus (adjusted OR 26.96, 95% CI 2.89-251.94), age over 50 years (adjusted OR 6.20, 95% CI 1.36-28.32) and thyroid dysfunction (adjusted OR 9.30, 95% CI 1.36-63.66). A fourth factor, baseline endogenous creatinine clearance below normal limits, was significant at the P = 0.05 level (adjusted OR 6.49, 95% CI 0.98-43.01). Hyperparathyroidism was noted in three cases of chronic poisoning suffering severe neurotoxicity. Severe lithium neurotoxicity occurs almost exclusively in the context of chronic therapeutic administration of lithium, and rarely results from acute ingestion of lithium, even in patients currently taking lithium. As such it is an iatrogenic illness, occurring in patients who have identifiable clinical risk factors: nephrogenic diabetes insipidus, older age, abnormal thyroid function and impaired renal function. Although administration of drugs which impair lithium clearance appeared to contribute minimally to chronic lithium poisoning in the absence of other factors, these drugs may well 'uncover' the predisposing risk factors and certainly should not be considered safe to use as a consequence of this study. The serious morbidity suffered by lithium toxic patients, and the cost to society due to long hospital stays, might be reduced by careful prescribing, vigilant monitoring and awareness of these factors, as they develop in otherwise stable patients. Review of existing therapeutic guidelines may be warranted.


A Transmural Path Model Improves the Definition of the Orthotropic Tissue Structure in Heart Simulations.
In the past decades, the structure of the heart, human as well as other species, has been explored in a detailed way, e.g., via histological studies or diffusion tensor magnetic resonance imaging. Nevertheless, the assignment of the characteristic orthotropic structure in a patient-specific finite element model remains a challenging task. Various types of rule-based models, which define the local fiber and sheet orientation depending on the transmural depth, have been developed. However, the correct assessment of the transmural depth is not trivial. Its accuracy has a substantial influence on the overall mechanical and electrical properties in rule-based models. The main purpose of this study is the development of a finite element-based approach to accurately determine the transmural depth on a general unstructured grid. Instead of directly using the solution of the Laplace problem as the transmural depth, we make use of a well-established model for the assessment of the transmural thickness. It is based on two hyperbolic first-order partial differential equations for the definition of a transmural path, whereby the transmural thickness is defined as the arc length of this path. Subsequently, the transmural depth is determined based on the position on the transmural path. Originally, the partial differential equations were solved via finite differences on structured grids. In order to circumvent the need of two grids and mapping between the structured (to determine the transmural depth) and unstructured (electromechanical heart simulation) grids, we solve the equations directly on the same unstructured tetrahedral mesh. We propose a finite-element-based discontinuous Galerkin approach. Based on the accurate transmural depth, we assign the local material orientation of the orthotropic tissue structure in a usual fashion. We show that this approach leads to a more accurate definition of the transmural depth. Furthermore, for the left ventricle, we propose functions for the transmural fiber and sheet orientation by fitting them to literature-based diffusion tensor magnetic resonance imaging data. The proposed functions provide a distinct improvement compared to existing rules from the literature.


Association between Free Triiodothyronine Levels and Peripheral Arterial Disease in Euthyroid Participants.
This current cross-sectional study investigates the relationship between thyroid hormones and peripheral artery disease (PAD) among euthyroid Chinese population aged 40 years and above. Serum free triiodothyronine (FT3), free thyroxin (FT4), thyroid-stimulating hormone (TSH), and thyroid antibodies were measured. PAD was defined as ankle-brachial index (ABI) < 0.9. There were 91 (2.9%) PAD cases among the 3,148 euthyroid study participants. Participants in the highest quartile of FT3 and free-triiodothyronine-to-free-thyroxin (FT3/FT4 ratio) had a decreased risk of prevalent PAD (multivariate-adjusted odds ratio, 95% confidence interval: 0.32, 0.15-0.62, P for trend = 0.01 and 0.31, 0.13-0.66, P for trend = 0.004, respectively) compared to those in the lowest quartile. To conclude, FT3 levels and the FT3/FT4 ratio was inversely associated with prevalent PAD in euthyroid Chinese population aged 40 years and above.