A single nomenclature and associated database for alleles at the major histocompatibility complex class II DRB1 locus of sheep.
The development of standardised nomenclatures with associated databases containing reference sequences for alleles at polymorphic loci within the major histocompatibility complex (MHC) has been facilitated by the development of the immuno polymorphism database (IPD). Recently, included within IPD-MHC is information on allelic diversity within sheep species (IPD-MHC-OLA). Here, we present the first report of progress in populating the sheep IPD-MHC database with alleles at the class II MHC DRB1 locus. The sequence of 63 Ovar-DRB1 alleles within 24 allelic families is now held within the database, each meeting the minimum requirement of a complete second exon. These sequences are derived from a combination of genomic and cDNA-based approaches and represent the most extensive collection of validated alleles at the sheep DRB1 locus yet described. Although these 63 alleles probably represent only a fraction of the DRB1 allelic diversity in sheep species worldwide, we encourage the research community to use the official allelic nomenclature and to contribute allelic sequences to the database via its web-based submission tool. In time, the IPD-MHC-OLA resource will underpin population-based MHC genotyping studies and help to simplify meta-analyses of multi-source data from wild and domestic sheep populations.


Space of symmetry matrices with elements 0, ±1 and complete geometric description; its properties and application.
A fixed set, that is the set of all lattice metrics corresponding to the arithmetic holohedry of a primitive lattice, is a natural tool for keeping track of the symmetry changes that may occur in a deformable lattice [Ericksen (1979). Arch. Rat. Mech. Anal. 72, 1-13; Michel (1995). Symmetry and Structural Properties of Condensed Matter, edited by T. Lulek, W. Florek & S. Walcerz. Singapore: Academic Press; Pitteri & Zanzotto (1996). Acta Cryst. A52, 830-838; and references quoted therein]. For practical applications it is desirable to limit the infinite number of arithmetic holohedries, and simplify their classification and construction of the fixed sets. A space of 480 matrices with cyclic consecutive powers, determinant 1, elements from {0, ±1} and geometric description were analyzed and offered as the framework for dealing with the symmetry of reduced lattices. This matrix space covers all arithmetic holohedries of primitive lattice descriptions related to the three shortest lattice translations in direct or reciprocal spaces, and corresponds to the unique list of 39 fixed points with integer coordinates in six-dimensional space of lattice metrics. Matrices are presented by the introduced dual symbol, which sheds some light on the lattice and its symmetry-related properties, without further digging into matrices. By the orthogonal lattice distortion the lattice group-subgroup relations are easily predicted. It was proven and exemplified that new symbols enable classification of lattice groups on an absolute basis, without metric considerations. In contrast to long established but sophisticated methods for assessing the metric symmetry of a lattice, simple filtering of the symmetry operations from the predefined set is proposed. It is concluded that the space of symmetry matrices with elements from {0, ±1} is the natural environment of lattice symmetries related to the reduced cells and that complete geometric characterization of matrices in the arithmetic holohedry provides a useful tool for solving practical lattice-related problems, especially in the context of lattice deformation.


Benefit of oral zinc supplementation as an adjunct to zidovudine (AZT) therapy against opportunistic infections in AIDS.
Zinc is perhaps the most important trace element for immune function. Congenital or acquired zinc deficiencies are associated with immune abnormalities and increased susceptibility to infectious diseases. AIDS subjects suffer from reduced zinc bioavailability, more severe in stage IV than in stage III. Such zinc deficiency causes, among other effects, a profound reduction in the biological activity of one of the thymic hormones, thymulin (zinc-facteur-timique-serique, ZnFTS). With these premises, zinc sulphate was administered orally at a daily dose of 200 mg for 30 days to AZT-treated stage III subjects with generalized lymphadenopathy (17 subjects) and stage IV subgroup C1 (12 subjects) AIDS patients. 18 stage III subjects with generalized lymphoadenopathy and 10 stage IV subgroup C1 subjects treated only with AZT served as controls. Zinc sulphate supplementation of stage III and in stage IV C1 patients was followed by an increase or a stabilization in the body weight and an increase of the number of CD4+ cells and the plasma level of active zinc-bound thymulin. The frequency of opportunistic infectious episodes in the 24 months following entry into the study was reduced after zinc supplementation in stage IV C1 subjects (11 infections vs 25 in controls) and delayed in stage III zinc-treated subjects (1 infection/24 months vs 13 infections/24 months in controls). The effect of zinc on opportunistic infections is restricted to infections due to Pneumocystis carinii and Candida, whereas no variations have been observed in the frequencies of cytomegalovirus and toxoplasma infections. These data may support the benefit of zinc as an adjunct to AZT therapy in AIDS pathology.


Technique for targeting arteriovenous malformations using frameless image-guided robotic radiosurgery.
To integrate three-dimensional (3D) digital rotation angiography (DRA) and two-dimensional (2D) digital subtraction angiography (DSA) imaging into a targeting methodology enabling comprehensive image-guided robotic radiosurgery of arteriovenous malformations (AVMs). DRA geometric integrity was evaluated by imaging a phantom with embedded markers. Dedicated DSA acquisition modes with preset C-arm positions were configured. The geometric reproducibility of the presets was determined, and its impact on localization accuracy was evaluated. An imaging protocol composed of anterior-posterior and lateral DSA series in combination with a DRA run without couch displacement between acquisitions was introduced. Software was developed for registration of DSA and DRA (2D-3D) images to correct for: (a) small misalignments of the C-arm with respect to the estimated geometry of the set positions and (b) potential patient motion between image series. Within the software, correlated navigation of registered DRA and DSA images was incorporated to localize AVMs within a 3D image coordinate space. Subsequent treatment planning and delivery followed a standard image-guided robotic radiosurgery process. DRA spatial distortions were typically smaller than 0.3 mm throughout a 145-mm × 145-mm × 145-mm volume. With 2D-3D image registration, localization uncertainties resulting from the achievable reproducibility of the C-arm set positions could be reduced to about 0.2 mm. Overall system-related localization uncertainty within the DRA coordinate space was 0.4 mm. Image-guided frameless robotic radiosurgical treatments with this technique were initiated. The integration of DRA and DSA into the process of nidus localization increases the confidence with which radiosurgical ablation of AVMs can be performed when using only an image-guided technique. Such an approach can increase patient comfort, decrease time pressure on clinical and technical staff, and possibly reduce the number of cerebral angiograms needed for a particular patient.


Alkali Metal- and Acid-Catalyzed Interconversion of Goniodomin A with Congeners B and C.
Goniodomin A (GDA, 1) is a phycotoxin produced by at least four species of Alexandrium dinoflagellates that are found globally in brackish estuaries and lagoons. It is a linear polyketide with six oxygen heterocyclic rings that is cyclized into a macrocyclic structure via lactone formation. Two of the oxygen heterocycles in 1 comprise a spiro-bis-pyran, whereas goniodomin B (GDB) contains a 2,7-dioxabicyclo[3.3.1]nonane ring system fused to a pyran. When H2O is present, 1 undergoes facile conversion to isomer GDB and to an α,β-unsaturated ketone, goniodomin C (GDC, 7). GDB and GDC can be formed from GDA by cleavage of the spiro-bis-pyran ring system. GDA, but not GDB or GDC, forms a crown ether-type complex with K+. Equilibration of GDA with GDB and GDC is observed in the presence of H+ and of Na+, but the equilibrated mixtures revert to GDA upon addition of K+. Structural differences have been found between the K+ and Na+ complexes. The association of GDA with K+ is strong, while that with Na+ is weak. The K+ complex has a compact, well-defined structure, whereas Na+ complexes are an ill-defined mixture of species. Analyses of in vitro A. monilatum and A. hiranoi cultures indicate that only GDA is present in the cells; GDB and GDC appear to be postharvest transformation products.


Natural allelic variation of the AZI1 gene controls root growth under zinc-limiting condition.
Zinc is an essential micronutrient for all living organisms and is involved in a plethora of processes including growth and development, and immunity. However, it is unknown if there is a common genetic and molecular basis underlying multiple facets of zinc function. Here we used natural variation in Arabidopsis thaliana to study the role of zinc in regulating growth. We identify allelic variation of the systemic immunity gene AZI1 as a key for determining root growth responses to low zinc conditions. We further demonstrate that this gene is important for modulating primary root length depending on the zinc and defence status. Finally, we show that the interaction of the immunity signal azelaic acid and zinc level to regulate root growth is conserved in rice. This work demonstrates that there is a common genetic and molecular basis for multiple zinc dependent processes and that nutrient cues can determine the balance of growth and immune responses in plants.


Automated classification of optical coherence tomography images for the diagnosis of oral malignancy in the hamster cheek pouch.
Most studies evaluating the potential of optical coherence tomography (OCT) for the diagnosis of oral cancer are based on visual assessment of OCT B-scans by trained experts. Human interpretation of the large pool of data acquired by modern high-speed OCT systems, however, can be cumbersome and extremely time consuming. Development of image analysis methods for automated and quantitative OCT image analysis could therefore facilitate the evaluation of such a large volume of data. We report automated algorithms for quantifying structural features that are associated with the malignant transformation of the oral epithelium based on image processing of OCT data. The features extracted from the OCT images were used to design a statistical classification model to perform the automated tissue diagnosis. The sensitivity and specificity of distinguishing malignant lesions from benign lesions were found to be 90.2% and 76.3%, respectively. The results of the study demonstrate the feasibility of using quantitative image analysis algorithms for extracting morphological features from OCT images to perform the automated diagnosis of oral malignancies in a hamster cheek pouch model.


Regulation of gene expression in bovine blastocysts in response to oxygen and the iron chelator desferrioxamine.
Low (2%) oxygen conditions during postcompaction culture of bovine blastocysts improve embryo quality and are associated with small increases in the expression of glucose transporter 1 (SLC2A1), anaphase promoting complex (ANAPC1), and myotrophin (MTPN), suggesting a role for oxygen in the regulation of embryo development, mediated through oxygen-sensitive gene expression. However, bovine embryos, to at least the blastocyst stage, lack detectable levels of the key regulator of oxygen-sensitive gene expression, hypoxia-inducible 1 alpha (HIF1A), while the less well-characterized HIF2 alpha protein is readily detectable. Here we report that other key HIF1 regulated genes are not significantly altered in their expression pattern in bovine blastocysts in response to reduced oxygen concentrations postcompaction-with the exception of lactate dehydrogenase A (LDHA), which was significantly increased following 2% oxygen culture. Antioxidant enzymes have been suggested as potential HIF2 target genes, but their expression was not altered following low-oxygen culture in the bovine blastocyst. The addition of desferrioxamine (an iron chelator and inducer of HIF-regulated gene expression) during postcompaction stages significantly increased SLC2A1, LDHA, inducible nitric oxide synthase (NOS2A), and MTPN gene expression in bovine blastocysts, although development to the blastocyst stage was not significantly affected. These results further suggest that expression of genes, known to be regulated by oxygen via HIF-1 in somatic cells, is not influenced by oxygen during preimplantation postcompaction bovine embryo development. Oxygen-regulated expression of LDHA and SLC2A1 in bovine blastocysts suggests that regulation of these genes may be mediated by HIF2. Furthermore, the effect of a reduced-oxygen environment on gene expression can be mimicked in vitro through the use of desferrioxamine. These results further support our data that the bovine blastocyst stage embryo is unique in its responsiveness to oxygen compared with somatic cells, in that the lack of HIF1-mediated gene expression reduces the overall response to low (physiological) oxygen environments, which appear to favor development.


Synthesis and release of luteinizing hormone in vitro: manipulations of Ca2+ environment.
We determined if luteinizing hormone (LH) synthesis is Ca2+ dependent and coupled to LH release. We monitored LH synthesis when LH release was stimulated either by specific [gonadotropin-releasing hormone (GnRH)] or nonspecific stimuli (50 mM K+ and 2 or 20 microM Ca2+ ionophore A23187) and inhibited by Ca2+-reduced medium. LH synthesis was estimated by measuring incorporation of [3H]glucosamine (glycosylation) and [14C]alanine (translation) into total (cell and medium) immunoprecipitable LH by cultured rat anterior pituitary cells. Both GnRH (1 nM) and 50 mM K+ significantly stimulated LH release and glycosylation, but had no effect on LH translation. A23187 also stimulated LH release, but significantly depressed glycosylation of LH and total protein and [14C]alanine uptake. Deletion of Ca2+ from the medium depressed both GnRH-induced LH release and glycosylation. Addition of 0.1 mM EGTA to Ca2+-free medium not only inhibited GnRH-induced release and glycosylation of LH but also uptake of precursors and glycosylation and translation of total protein. Thus glycosylation and release of LH are Ca2+ dependent. Whether parallel changes in LH release and glycosylation reflect a cause and effect relationship remains to be determined.


An Aversive Response to Osmotic Upshift in Caenorhabditis elegans.
Environmental osmolarity presents a common type of sensory stimulus to animals. While behavioral responses to osmotic changes are important for maintaining a stable intracellular osmolarity, the underlying mechanisms are not fully understood. In the natural habitat of Caenorhabditis elegans, changes in environmental osmolarity are commonplace. It is known that the nematode acutely avoids shocks of extremely high osmolarity. Here, we show that C. elegans also generates gradually increased aversion of mild upshifts in environmental osmolarity. Different from an acute avoidance of osmotic shocks that depends on the function of a transient receptor potential vanilloid channel, the slow aversion to osmotic upshifts requires the cGMP-gated sensory channel subunit TAX-2. TAX-2 acts in several sensory neurons that are exposed to body fluid to generate the aversive response through a motor network that underlies navigation. Osmotic upshifts activate the body cavity sensory neuron URX, which is known to induce aversion upon activation. Together, our results characterize the molecular and cellular mechanisms underlying a novel sensorimotor response to osmotic stimuli and reveal that C. elegans engages different behaviors and the underlying mechanisms to regulate responses to extracellular osmolarity.


Beyond osteogenesis: an in vitro comparison of the potentials of six bone morphogenetic proteins.
Bone morphogenetic proteins (BMPs) other than the clinically available BMP-2 and BMP-7 may be useful for improving fracture healing through both increasing osteogenesis and creating a favorable healing environment by altering cytokine release by endogenous cells. Given the spectrum of potential applications for BMPs, the objective of this study was to evaluate various BMPs under a variety of conditions to provide further insight into their therapeutic capabilities. The alkaline phosphatase (ALP) activity of both C2C12 and human adipose-derived stem cells (hASCs) was measured after exposure of increasing doses of recombinant human BMP-2, -4, -5, -6, -7, or -9 for 3 and 7 days. BMPs-2, -4, -5, -6, -7, and -9 were compared in terms of their ability to affect the release of stromal derived factor-1 (SDF-1), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (b-FGF) from human bone marrow stromal cells (hBMSCs). Gene expression of ALP, osteocalcin, SDF-1, VEGF, and b-FGF following shRNA-mediated knockdown of BMP-2 and BMP-6 in hBMSCs or human osteoblasts under osteogenic differentiation conditions was also evaluated. Collectively, BMPs-6 and -9 produced the greatest osteogenic differentiation of C2C12 and hASCs as determined by ALP. The hBMSC secretion of SDF-1 was most affected by BMP-5, VEGF by BMP-4, and b-FGF by BMP-2. The knockdown of BMP-2 in BMSCs had no effect on any of the genes measured whereas BMP-6 knockdown in hBMSCs caused a significant increase in VEGF gene expression. BMP-2 and BMP-6 knockdown in human osteoblasts caused significant increases in VEGF gene expression and trends toward decreases in osteocalcin expression. These findings support efforts to study other BMPs as potential bone graft supplements, and to consider combined BMP delivery for promotion of multiple aspects of fracture healing.


Venous thromboembolism prophylaxis with unfractionated heparin in the hospitalized medical patient: the case for thrice daily over twice daily dosing.
For venous thromboembolism (VTE) prevention in the hospitalized medical patient, no head-to-head trials have been performed of unfractionated heparin (UFH) 5,000 U subcutaneously thrice (i.e. q8 h or TID) daily versus twice daily (q12 h or BID). Several meta-analyses have been undertaken in attempts to determine whether one regimen may be more beneficial for safety and efficacy. Currently, not all international guidelines include a recommended frequency for UFH. Delineation of this frequency may be helpful to the practicing clinician. Primary studies (with a modified Jadad score of >or=6 to demonstrate a stronger study design) that compared low molecular weight heparin (LMWH) and UFH, and UFH and placebo were evaluated. Meta-analyses evaluating safety and efficacy of LMWH versus UFH, or TID UFH versus BID UFH were also evaluated. Although BID UFH shows some efficacy in one primary study, it is no more beneficial than no prophylaxis in another study. LMWH appears to be more efficacious than BID UFH, but comparable in safety and efficacy to TID UFH. Meta-analytic data demonstrates that BID UFH may have some reduction in deep vein thrombosis. Meta-analytic data also suggests that TID UFH is more efficacious than BID UFH at the cost of more major bleeding. The medical patient with risk factors for the development of VTE appears to be at moderate to high risk. International guidelines for VTE prevention should incorporate a frequency for UFH to guide use. TID UFH is superior in efficacy to BID UFH even when taking into consideration the increased rate of major bleeds. Newly published risk-assessment models may be beneficial in determining which patients would best benefit from BID UFH or TID UFH.


Cannabis use and neurocognitive performance at 13-14 Years-Old: Optimizing assessment with hair toxicology in the Adolescent brain cognitive development (ABCD) study.
Cannabis is widely used, including in early adolescence, with prevalence rates varying by measurement method (e.g., toxicology vs. self-report). Critical neurocognitive development occurs throughout adolescence. Given conflicting prior brain-behavior results in cannabis research, improved measurement of cannabis use in younger adolescents is needed. Data from the Adolescent Brain Cognitive Development (ABCD) Study Year 4 follow-up (participant age: 13-14 years-old) included hair samples assessed by LC-MS/MS and GC-MS/MS, quantifying THCCOOH (THC metabolite), THC, and cannabidiol concentrations, and the NIH Toolbox Cognitive Battery. Youth whose hair was positive for cannabinoids or reported past-year cannabis use were included in a Cannabis Use (CU) group (n = 123) and matched with non-using Controls on sociodemographics (n = 123). Standard and nested ANCOVAs assessed group status predicting cognitive performance, controlling for family relationships. Follow-up correlations assessed cannabinoid hair concentration, self-reported cannabis use, and neurocognition. CU scored lower on Picture Memory (p = .03) than Controls. Within the CU group, THCCOOH negatively correlated with Picture Vocabulary (r = -0.20, p = .03) and Flanker Inhibitory Control and Attention (r = -0.19, p = .04), and past-year cannabis use was negatively associated with List Sorting Working Memory (r = -0.33, p = .0002) and Picture Sequence Memory (r = -0.19, p = .04) performances. Youth who had used cannabis showed lower scores on an episodic memory task, and more cannabis use was linked to poorer performances on verbal, inhibitory, working memory, and episodic memory tasks. Combining hair toxicology with self-report revealed more brain-behavior relationships than self-report data alone. These youth will be followed to determine long-term substance use and neurocognition trajectories.


Online learning versus workshops: a rank minimized trial comparing the effect of two knowledge translation strategies designed to alter knowledge, readiness to change, and self-efficacy with respect to rehabilitation outcome measures.
Traditional face-to-face learning is often replaced by virtual learning because it can be more feasible and cost-effective, and more recently due to the need for social distancing. The objective was to evaluate the effectiveness of two innovative knowledge translation (KT) interventions; in-person stakeholder-hosted, interactive, problem-based seminars (SHIPS) versus online problem-based tutorials (e-PBL) in changing knowledge, readiness to change, and self-efficacy with respect to the use of rehabilitation outcome measures. Physical and occupational therapists (n = 124) were recruited from four sites across Canada to participate in either an e-PBL or SHIPS. Evaluations of KT impact measured knowledge, self-efficacy to implement outcome measures in practice, and readiness to change. There were 112 participants who completed the study. Following the intervention, the mean knowledge scores for both groups improved, but there was greater improvement in participants who completed SHIPS. For self-efficacy and readiness to change, there was no significant difference between groups six months following the interventions. E-PBL was as good as a SHIPS for improving and retaining self-efficacy and readiness to change. Knowledge improved more with workshops than online delivery, while improvements in self-efficacy and readiness to change improved similarly regardless of delivery.Implications for RehabilitationThis study compared the relative efficacy of internet and workshop-based education, focusing specifically on the use of outcome measures in physical and occupational therapy practice.Improvements in the self-efficacy of rehabilitation professionals with respect to outcome measure use had lasting effects regardless of KT intervention type, as it was retained six months following the intervention.Results from this study show that online interventions may be as effective as face-to-face workshops for improving readiness to change and self-efficacy for using outcome measures in practice by rehabilitation professionals.This is valuable information given the recent global pandemic, the need for social distancing, and the potential for learning interventions to focus within the online environment in the future.


Prospective, double-blind, randomized clinical trial comparing an ERAS pathway with ketorolac and pregabalin versus standard of care plus placebo during live donor nephrectomy for kidney transplant.
Opioid exposure is a concern after live donation for kidney transplant. We theorized that an enhanced recovery after surgery pathway (ERAS) using pregabalin preoperatively to desensitize nerves followed by the nonsteroidal anti-inflammatory drug ketorolac, during and after surgery, can control pain, thus requiring less perioperative narcotics. The aim of this study was to determine if the use of a nonopioid analgesic ERAS protocol for donor nephrectomies could decrease the use of narcotics without an increase in complications compared with standard of care (SOC). This is a single-center, prospective, double-blind, randomized clinical trial involving a total of 62 patients undergoing nephrectomy for live donor kidney transplant. Length of hospital stay (LOS) was significantly reduced by 10% in the ERAS group versus the SOC-plus-placebo group. Morphine dose equivalents were significantly reduced by 40% in the study group versus the SOC-plus-placebo group. The use of this nonopioid analgesic ERAS pathway for donor nephrectomies decreased the use of narcotics without an increase in complications compared with SOC. There was significantly reduced LOS and less narcotic use in the study group versus the SOC-plus-placebo group. (ClinicalTrials.gov registration number: NCT03669081).


Regulation of postsynaptic function by the dementia-related ESCRT-III subunit CHMP2B.
The charged multivesicular body proteins (Chmp1-7) are an evolutionarily conserved family of cytosolic proteins that transiently assembles into helical polymers that change the curvature of cellular membrane domains. Mutations in human CHMP2B cause frontotemporal dementia, suggesting that this protein may normally control some neuron-specific process. Here, we examined the function, localization, and interactions of neuronal Chmp2b. The protein was highly expressed in mouse brain and could be readily detected in neuronal dendrites and spines. Depletion of endogenous Chmp2b reduced dendritic branching of cultured hippocampal neurons, decreased excitatory synapse density in vitro and in vivo, and abolished activity-induced spine enlargement and synaptic potentiation. To understand the synaptic effects of Chmp2b, we determined its ultrastructural distribution by quantitative immuno-electron microscopy and its biochemical interactions by coimmunoprecipitation and mass spectrometry. In the hippocampus in situ, a subset of neuronal Chmp2b was shown to concentrate beneath the perisynaptic membrane of dendritic spines. In synaptoneurosome lysates, Chmp2b was stably bound to a large complex containing other members of the Chmp family, as well as postsynaptic scaffolds. The supramolecular Chmp assembly detected here corresponds to a stable form of the endosomal sorting complex required for transport-III (ESCRT-III), a ubiquitous cytoplasmic protein complex known to play a central role in remodeling of lipid membranes. We conclude that Chmp2b-containing ESCRT-III complexes are also present at dendritic spines, where they regulate synaptic plasticity. We propose that synaptic ESCRT-III filaments may function as a novel element of the submembrane cytoskeleton of spines.


Mutant of Bordetella pertussis which lacks ability to produce filamentous hemagglutinin.
Mutant cultures that lacked the ability to produce filamentous hemagglutinin were obtained by UV irradiation of Bordetella pertussis strain 18-323. Various biological activities of these mutants were compared with those of the parent culture. It was found that the leukocytosis-promoting, histamine-sensitizing, and mouse-protecting activities of the mutants were similar to those of the parent strain but that the virulence to mice was lower. The results seemed to rule out fimbrial hemagglutinin as a protective antigen when the ability to protect mice was measured by the intracerebral challenge test.


A measure of explained variation for event history data.
There is no shortage of proposed measures of prognostic value of survival models in the statistical literature. They come under different names, including explained variation, correlation, explained randomness, and information gain, but their goal is common: to define something analogous to the coefficient of determination R(2) in linear regression. None however have been uniformly accepted, none have been extended to general event history data, including recurrent events, and many cannot incorporate time-varying effects or covariates. We present here a measure specifically tailored for use with general dynamic event history regression models. The measure is applicable and interpretable in discrete or continuous time; with tied data or otherwise; with time-varying, time-fixed, or dynamic covariates; with time-varying or time-constant effects; with single or multiple event times; with parametric or semiparametric models; and under general independent censoring/observation. For single-event survival data with neither censoring nor time dependency it reduces to the concordance index. We give expressions for its population value and the variance of the estimator and explore its use in simulations and applications. A web link to R software is provided.


Assessing the Readiness of Nursing Sectors in Low- and Middle-Income Countries to Adopt Holistic Practice: Rwanda as Exemplar.
Over the past several years, holistic nursing education has become more readily available to nurses working in high-income nations, and holistic practice has become better defined and promoted through countless organizational and governmental initiatives. However, global nursing community members, particularly those serving in low- and middle-income countries (LMICs) within resource-constrained health care systems, may not find holistic nursing easily accessible or applicable to practice. The purpose of this article is to assess the readiness of nursing sectors within these resource-constrained settings to access, understand, and apply holistic nursing principles and practices within the context of cultural norms, diverse definitions of the nursing role, and the current status of health care in these countries. The history, current status, and projected national goals of professional nursing in Rwanda is used as an exemplar to forward the discussion regarding the readiness of nurses to adopt holistic education into practice in LMICs. A background of holistic nursing practice in the United States is provided to illustrate the multifaceted aspects of support necessary in order that such a specialty continues to evolve and thrive within health care arenas and the communities it cares for.


Merkel cell carcinoma of the eyelid and orbit.
The incidence of Merkel cell carcinoma has tended to increase worldwide in recent years. Merkel cell carcinoma is a rare tumor of the skin that occurs mainly in the sun exposed sites. The malignant reversal of Merkel cells is currently associated with an infection caused by a Merkel cell polyomavirals. In some cases, the disease may have a relatively inconspicuous clinical picture in the initial phase, which is in contrast to its extensive microscopic propagation. For this reason, the risk of late diagnosis or insufficient primary surgery is increased. The diagnostic standard is histological and, in particular, immunohistochemical examination of tumor tissue samples. Merkel cell carcinoma is a marked tendency to local recurrence and early development of metastases in regional lymph nodes, followed by generalization. The basis of treatment is radical excision of the tumor by in most cases by adjuvant radiotherapy targeted at primary place of occurrence and the area of regional draining lymph nodes. The effectiveness of different chemotherapeutic protocols in Merkel cell carcinoma is mostly low and the median survival is low. From a prognostic point of view, Merkel cell carcinoma plays the most important role of staging the tumor at the time of capture. The suspected lesions in the area around the eye, eyelid and orbit need to indicate adequate therapeutic approach that the detection of the disease at the earliest stage. The authors describe the clinical experience in 2 patients with Merkel cell carcinoma of the eyelid and orbit. Key words: eyelid tumors, tumors of the orbit, Merkel cell carcinoma.


Toward imaging biomarkers for osteoarthritis.
Many new therapeutic strategies have been and are being developed to correct, prevent, or slow the progression of osteoarthritis. Our ability to evaluate the efficacy of these techniques, or to determine the situations for which they might provide the most benefit, critically depends on diagnostic measures that can serve as proxies for the present or predicted state of the cartilage. Many of the magnetic resonance imaging techniques that have been emerging over the past decades appear promising in that they have shown technical validity in measuring the morphologic and molecular state of cartilage. With continued development and added insight from pilot clinical studies, these or related methods may soon be in customary use. These techniques are part of a paradigm shift where therapeutic strategies are developed hand-in-hand with diagnostic approaches-a shift that offers the promise of speeding development of effective therapies, and focusing their use in areas where they can be most successful.


T4 late transcripts are initiated near a conserved DNA sequence.
Bacteriophage T4 late transcription is unusual, among prokaryotes, in its complexity. Late transcription requires the host RNA polymerase, the products of T4 genes, 33, 45 and 55, and other small polypeptides, the genes of which have not been identified. In addition the DNA template must be "competent' for late transcription. First the DNA must contain the substituted base 5-hydroxymethyl cytosine in place of cytosine (this requirement is eliminated by a mutation in the T4 alc gene). Second, the DNA must be replicating, although late transcription can be uncoupled from DNA replication by mutations in the T4 genes coding for DNA ligase (gene 30) and DNA exonuclease (gene 46). We report here the location of the initiation sites of the messenger RNAs (mRNAs) synthesized in vivo from four late genes (genes 21, 22, 23 and 36) by S1 nuclease mapping and we have determined the DNA sequences at these sites. We have found strong homology to the sequence TATAAATACTATT immediately upstream from the 5' ends of the late messages and we suggest that this sequence is specifically recognized by the complex responsible for late transcription. Also, we have examine gene 23 mRNA synthetized in the absence of DNA replication using the 30- 46- mutant described above and find that it is identical to the true late transcript synthesized in normal infections.


Natural protein engineering: a uniquely salt-tolerant, but not halophilic, alpha-type carbonic anhydrase from algae proliferating in low- to hyper-saline environments.
Dunaliella salina is a unicellular green alga thriving in environments ranging from fresh water to hyper-saline lakes, such as the Dead Sea. An unusual, internally duplicated, 60 kDa alpha-type carbonic anhydrase (dCA I), located on the surface of this alga, is expected to function over a broad range of salinities. It would therefore differ from other carbonic anhydrases that already lose activity at low salinities and also from halophilic proteins that require high salinities for conformational stability. Enzymatic analyses indeed indicated that dCA I retained activity at salt concentrations ranging from low salt to at least 1.5 M NaCl or KCl for CO(2) hydration, 2.0 M NaCl for esterase activity and 0.5 M for bicarbonate dehydration. Although measurements at higher salinities were constrained by the interference of salt in the respective assayed reactions, activity was noticeable even at 4.0 M NaCl. Comparisons of the internally duplicated dCA I to single-domain derivatives indicated that inter-domain interactions played a decisive role in the stability, activity, salt tolerance and pH responses of dCA I. Hence dCA I is a uniquely salt- tolerant protein, retaining an active conformation over a large range of salinities and, as a Zn metalloenzyme, largely immune to the specific inhibitory effects of anions. Its unique features make dCA I a useful model to understand the physico-chemical basis of halotolerance and protein-salt interactions in general.


A systematic review and meta-analysis of prospective associations between alcohol consumption and incident frailty.
light-to-moderate alcohol consumption is protective against all-cause mortality and cardiovascular diseases. There is limited evidence in the literature on how alcohol consumption is related to frailty. five databases (Embase, Scopus, MEDLINE, CINAHL, PsycINFO) were systematically searched in July 2016 for prospective studies published between 2000 and 2016 examining baseline alcohol consumption and subsequent frailty risk among middle-aged or older community-dwelling population. Odds ratios (ORs) for incident frailty were pooled using a random-effects model. Heterogeneity, methodological quality and publication bias were assessed. of 926 studies identified by the systematic search, four studies were included (total n = 44,051, ≥55 years, 66.2% alcohol users). OR of incident frailty for the highest (at least 24 g of alcohol/day for men, 12g of alcohol/day for women) or the most frequent (≥5 days of drinking/week) alcohol consumption compared with no drinking were used for a meta-analysis. Pooled OR among three studies measuring alcohol consumption quantitatively showed that the highest alcohol consumption was associated with lower frailty risk (3 studies:pooled OR = 0.44, 95%CI = 0.19-1.00, P = 0.05). Adding the other study measuring frequency of alcohol consumption made little change (4 studies:pooled OR = 0.61, 95%CI = 0.44-0.77, P < 0.001). Two of the included studies suggested a possible U-shaped association with lowest risks for moderate drinkers. Heterogeneity was moderate in both analyses (I2 = 52-67%). There was no evidence of publication bias. this systematic review and meta-analysis study provides the first pooled evidence suggesting that heavier alcohol consumption is associated with lower incident frailty compared with no alcohol consumption among community-dwelling middle-aged and older people. However, this association may be due to unadjusted effect measures, residual confounding, 'sick quitter' effect or survival bias.


Human immunodeficiency virus (HIV) infection, sexually transmitted diseases and HIV-antibody testing practices in Belgian prostitutes.
From December 1988 to April 1989, 154 female prostitutes in and around Ghent, Belgium, were interviewed about their knowledge, attitudes and practices in relation to the risks for sexually transmitted diseases (STD) and human immunodeficiency virus (HIV) infection in their profession. Thirty four women worked as window prostitutes, 120 picked up their clients in bars, clubs, and saunas. Blood samples were taken from 123 women. One (0.8%) was seropositive for HIV1, 19 (15.4%) had Hepatitis B core antibodies (anti-HBc), eight (6.4%) showed markers of syphilis. None of them were Hepatitis B surface antigen (HBsAg) carriers. Hepatitis C antibodies (anti-HCV) were present in the serum of three women (2.4%). Overall STD seroprevalence was higher in the group of window prostitutes than in the group of club prostitutes. One woman admitted intravenous drug use. Former testing for anti-HIV antibodies had been performed in 102 (66.5%) respondents, of whom 84 (82.3%) were tested in the year preceding the interview. In 74.5% of the cases, these tests were requested by the women themselves. These results suggest that HIV infection is not yet prevalent in non-intravenous drug using prostitutes in Ghent, but that this situation may change considering their higher rates of past STD. Window prostitutes are at higher risk than club prostitutes. Testing for HIV seems to be common practice, mostly at the request of the women themselves. Health education should discourage the notion of testing as an alternative to using condoms.


RNA synthesis initiates in vitro conversion of M13 DNA to its replicative form.
Soluble enzyme fractions from uninfected Escherichia coli convert M13 and varphiX174 viral single strands to their double-stranded replicative forms. Rifampicin, an inhibitor of RNA polymerase, blocks conversion of M13 single strands to the replicative forms in vivo and in vitro. However, rifampicin does not block synthesis of the replicative forms of varphiX174 either in vivo or in soluble extracts. The replicative form of M13 synthesized in vitro consists of a full-length, linear, complementary strand annealed to a viral strand. The conversion of single strands of M13 to the replicative form proceeds in two separate stages. The first stage requires enzymes, ribonucleoside triphosphates, and single-stranded DNA; the reaction is inhibited by rifampicin. The macromolecular product separated at this stage supports DNA synthesis with deoxyribonucleoside triphosphates and a fresh addition of enzymes; ribonucleoside triphosphates are not required in this second stage nor does rifampicin inhibit the reaction. We presume that in the first stage there is synthesis of a short RNA chain, which then primes the synthesis of a replicative form by a DNA polymerase.


Influence of oral microbiome on longitudinal patterns of oral mucositis severity in patients with squamous cell carcinoma of the head and neck.
This study investigated the influence of oral microbial features on the trajectory of oral mucositis (OM) in patients with squamous cell carcinoma of the head and neck. OM severity was assessed and buccal swabs were collected at baseline, at the initiation of cancer treatment, weekly during cancer treatment, at the termination of cancer treatment, and after cancer treatment termination. The oral microbiome was characterized via the 16S ribosomal RNA V4 region with the Illumina platform. Latent class mixed-model analysis was used to group individuals with similar trajectories of OM severity. Locally estimated scatterplot smoothing was used to fit an average trend within each group and to assess the association between the longitudinal OM scores and longitudinal microbial abundances. Four latent groups (LGs) with differing patterns of OM severity were identified for 142 subjects. LG1 has an early onset of high OM scores. LGs 2 and 3 begin with relatively low OM scores until the eighth and 11th week, respectively. LG4 has generally flat OM scores. These LGs did not vary by treatment or clinical or demographic variables. Correlation analysis showed that the abundances of Bacteroidota, Proteobacteria, Bacteroidia, Gammaproteobacteria, Enterobacterales, Bacteroidales, Aerococcaceae, Prevotellaceae, Abiotrophia, and Prevotella_7 were positively correlated with OM severity across the four LGs. Negative correlation was observed with OM severity for a few microbial features: Abiotrophia and Aerococcaceae for LGs 2 and 3; Gammaproteobacteria and Proteobacteria for LGs 2, 3, and 4; and Enterobacterales for LGs 2 and 4. These findings suggest the potential to personalize treatment for OM. Oral mucositis (OM) is a common and debilitating after effect for patients treated for squamous cell carcinoma of the head and neck. Trends in the abundance of specific microbial features may be associated with patterns of OM severity over time. Our findings suggest the potential to personalize treatment plans for OM via tailored microbiome interventions.


Biomonitoring: A tool to assess PFNA body burdens and evaluate the effectiveness of drinking water intervention for communities in New Jersey.
Elevated perfluorononanoic acid (PFNA) levels, one of many manmade per- and polyfluoroalkyl substances (PFAS), were detected in public water systems/private wells in New Jersey communities. Interventions to end exposure through drinking water were carried out from 2014 to 2016. To evaluate the effectiveness of interventions, a community biomonitoring study was conducted for the communities between 2017 and 2020. A convenience sampling design was used with 120 participants in Year 1 between ages of 20-74 who consumed PFNA-contaminated water. Three blood samples, one year apart, were drawn from each participant and completed for 99 participants. Separated serum samples were measured for 12 PFAS including PFNA. Questionnaires were administered to collect information on demographics and potential sources. Drinking water and house dust collected at the first visit were analyzed for 14 PFAS including PFNA. The PFNA sera levels (Year 1) found 84 out of 120 (70%) participants were higher than the 95th percentile of a nationally representative sample of US adults (NHANES2015-16). Current drinking water and house dust were not significant contributing sources for the study participants. On average, PFNA sera levels were 12 ± 16% (Year 2) and 27 ± 16% (Year 3) lower than the level measured in Year 1 (p < 0.01). The PFNA half-life was estimated around 3.52 years, using a mixed model from 68 high-exposed participants (>95th percentile of NHANES2015-16) with controlling for physiological covariates. The decline in adult serum PFNA levels seen in the years following a community drinking water intervention suggests the intervention effectively reduced PFNA exposure via drinking water.


Identification of multiple antigens recognized by tumor-infiltrating lymphocytes from a single patient: tumor escape by antigen loss and loss of MHC expression.
The authors describe a patient who experienced recurrence of metastatic melanoma after an initial dramatic response to immunotherapy using peptides derived from gp100, MART-1, and tyrosinase emulsified in incomplete Freund's adjuvant, and present data to support the hypothesis that the progression of disease in this patient was due to in vivo immunoselection for immunoresistant tumor variants. The authors previously demonstrated the existence of T-cell clones in this patient's peripheral blood and tumor-infiltrating lymphocytes (TILs) reactive against multiple antigens, including gp100, the tyrosinase-related protein (TRP)-2, a novel TRP-2 isoform-TRP-2-6b, SOX10, and the melanoma antigen NY-ESO-1. In addition to the multiple HLA-A2 restricted T-cell clones, the authors have now identified additional HLA-B/C-restricted as well as class II (HLA-DP)-restricted anti-melanoma antigen T-cell clones from this patient's TIL. One recurrent tumor showed loss of expression of multiple tumor antigens but retention of HLA class I expression. The other recurrent lesion showed total loss of HLA class I expression even though the tumor cells still expressed many melanoma antigens. This paper thus provides evidence for both the effectiveness of the immune destruction of cancer as well as problems associated with antigen-loss tumor escape mechanisms.


A Case of Life-threatening Amlodipine and Atenolol Overdose.
Treating a patient of amlodipine-atenolol poisoning is nightmare for a physician. In high dose both the drugs individually cause severe bradycardia and hypotension. In combination they cause severe cardiovascular depression. Here we report a case of 66-year-old obese, hypertensive, depressed male, who presented to emergency 9 hours after consumption of 25 tablets of amlodipine-atenolol (5 mg+50 mg). On evaluation, he had refractory bradycardia, hypotension and acute kidney injury (AKI). Eventually he developed cardiac arrest. He was revived after 5 minutes of cardio-pulmonary resuscitation (CPR). He was successfully managed with gastric lavage, fluids, inotropes, atropine, isoprenaline and subsequently with calcium gluconate infusion, high-dose insulin euglycemia therapy (HIET) and lipid emulsion therapy. Glucagon infusion was also planned but it was not available. Patient hemodynamics improved and on 8th day he got the discharge. Our case exemplifies the importance of timely and aggressive management of lethal overdose of amlodipine-atenolol poisoning. How to cite this article: Tale S, Kumar M, Ghosh S, Bhalla A. A Case of Life-threatening Amlodipine and Atenolol Overdose. Indian J Crit Care Med 2019;23(6):281-283.


IS26-mediated plasmid reshuffling results in convergence of toxin-antitoxin systems but loss of resistance genes in XDR Klebsiella pneumoniae from a chronic infection.
Carbapenem-resistant Enterobacterales pose an urgent threat to human health worldwide. Klebsiella pneumoniae sequence type (ST) 14, initially identified in the Middle East and South-Asia and co-harbouring the carbapenemase genes bla OXA-232 and bla NDM-1, is now emerging globally. One such strain was detected in the USA in 2013 from a patient initially treated in India that also carried armA, a 16S rRNA methyltransferase that confers resistance to all clinically relevant aminoglycosides. Genetic and phenotypic changes were observed in 14 serial isolates collected from this chronically infected patient. The index isolate carried five plasmids, including an IncFIB-IncHI1B (harbouring armA and bla NDM-1), an IncFIA (bla CTX-M-15) and a ColE-like (bla OXA-232), and was extensively resistant to antibiotics. Four years later, a subsequent isolate had accumulated 34 variants, including a loss-of-function mutation in romA, resulting in tigecycline non-susceptibility. Importantly, this isolate now only carried two plasmids, including a large mosaic molecule made of fragments, all harbouring distinct toxin-antitoxin systems, from three of the canonical plasmids. Of the original acquired antibiotic resistance genes, this isolate only retained bla CTX-M-15, and as a result susceptibility to the carbapenems and amikacin was restored. Long-read sequencing of a subset of five representative isolates, collected between 2013 and 2017, allowed for the elucidation of the complex plasmid patterns and revealed the role of IS26-mediated plasmid reshuffling in the evolution of this clone. Such investigations of the mechanisms underlying plasmid stability, together with global and local surveillance programmes, are key to a better understanding of plasmid host range and dissemination.


[Initial indices for calculating the integral criteria of the effectiveness of the enzyme immobilization procedure].
The possibility of using a simplified model of the kinetics of the enzymatic process catalyzed by the immobilized enzyme for definition of the integral criteria for the efficacy of the immobilization process is theoretically validated. The means for determination of the initial characteristics for these criteria, i.e. the constants of the inactivation rate of the immobilized enzyme and the critical value of the catalyst activity are discussed. The use of the catalyst under the manufacturing conditions at the value lower than the critical one is not advisable.


Care System Versus Transmitted Light Wavefront Pattern of Contact Lenses.
This article compares the optical performance of soft contact lenses (CLs) treated with multipurpose or hydrogen peroxide care systems. The investigated care systems were (1) 3% hydrogen peroxide solution Oxysept (Abbot Medical Optics, Abbott Park, IL) and (2) multipurpose solution Regard (Vita Research, Ariccia, Italy). Three types of silicone hydrogel CLs were studied (comfilcon A, lotrafilcon B, and balafilcon A), unworn and exposed for 30 times to the solutions, which were replaced every 8 hr. The optical performance of the CLs was evaluated through the on-eye transmitted light wavefront patterns by considering new CLs as references. The surface morphology of the CLs was investigated by scanning electron microscopy. Statistically significant modifications in the range 0.1 to 0.3 μm of Zernicke coefficients and modifications of the root mean square of the wavefront aberration function were found for CLs treated with multipurpose solution, in agreement with the observed modifications of the surface morphology. Statistically significant changes were also found after exposure to the hydrogen peroxide solution, but the variation of the Zernicke coefficients was found lower than 0.1 μm, thus being negligible in CL optical performances. In addition to disinfection ability and ocular surface reactions, CL care systems are different in solution-related CL optical performance. Multipurpose solutions may affect the CL surface morphology with significant modifications of the transmitted light wavefront pattern.


Regulated proteolysis as an element of ER stress and autophagy: Implications for intestinal inflammation.
Endoplasmic reticulum (ER) stress and autophagy are tightly controlled cellular processes, which are responsible for maintaining protein homeostasis in a cell. Impairment of the interlinking pathways have been implicated in a number of human diseases, prominently in inflammatory bowel disease, where genetic variants in several independent autophagy and ER stress related loci have been associated to increased disease risk. Autophagy is a selective quality control process, which governs the integrity of the cell by removal of aged organelles and proteins via the lysosome, but recently has been shown to actively license the outcome of other signaling pathways by guiding the proteolytic removal of signaling protein complexes (adaptophagy). In this review, we summarize our knowledge on regulated proteolytic events involved in ER stress responses and autophagy, their interplay and potential regulatory effects with a particular focus on intestinal inflammation. This article is part of a Special Issue entitled: Proteolysis as a Regulatory Event in Pathophysiology edited by Stefan Rose-John.


Increased Serum Soluble Interleukin-2 Receptor Associated with Severity of Acute Exacerbation of Chronic Obstructive Pulmonary Disease.
This study aimed to reveal the correlation between serum soluble interleukin-2 receptor (sIL-2R) and prognosis in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). A total of 315 patients diagnosed with AECOPD between December 2017 and June 2020 were enrolled. The patients were divided into the good and adverse groups based on the outcomes. An adverse outcome in COPD exacerbation was defined by the presence of at least one of the following: (1) death from a respiratory cause during hospitalisation or within 1 month of follow-up; (2) intensive care unit admission; (3) invasive or non-invasive mechanical ventilation; and (4) COPD-related emergency visit or readmission within 1 month of follow-up. A good outcome was considered as the absence of all the aforementioned issues. The patients underwent lung function (spirometry) assessment, and clinical and inflammatory profiles were collected. Univariate and multivariate analyses were performed to identify the correlation between serum sIL-2R concentration and other variables related to adverse outcomes of AECOPD. The receiver operating characteristic curve was used to show the predictive ability of sIL-2R for adverse outcomes of AECOPD. We enrolled 315 patients, of whom 161 and 154 had good and adverse outcomes, respectively. We demonstrated that patients with adverse outcomes of AECOPD had a higher concentration of serum sIL-2R than patients with good outcomes (p < 0.001). The increased serum sIL-2R was positively associated with mMRC scores (p < 0.001), GLOD grades (p < 0.001), frequent exacerbation (p < 0.001), and smoking (p < 0.001) in patients with AECOPD and negatively correlated with pulmonary function (p < 0.001). An elevated sIL-2R level was a predictor for the risk of adverse outcomes in AECOPD with a cut-off value of 860 U/mL. Increased serum sIL-2R concentration correlated with the risk of the adverse outcomes in AECOPD, indicating that it can be a predictive factor contributing to the diagnosis and assessment of adverse outcomes in patients with AECOPD.


Self-monitoring dysfunction and the schizophrenic symptoms of alien control.
Frith & Done (1988) have proposed that the experience of alien control symptoms in schizophrenia is related to a failure by such individuals to monitor effectively their own willed intentions, actions and thoughts. To examine this hypothesis, a heterogeneous group of 35 patients, all carrying a DSM-III-R diagnosis of schizophrenia (or schizophreniform psychosis) and 24 non-patient controls, completed a battery of neuropsychological and cognitive tests, which inter alia, included four putative measures of self-monitoring. Patients took part in a detailed clinical interview to assess current levels of symptomatology. Patients generally performed at a lower level on most components of the test battery, including the four self-monitoring tests. Moreover, patients currently experiencing symptoms of alien control tended to experience greater difficulty with each of the self-monitoring tests; an effect that was relatively independent of neuropsychological or general cognitive function. The relationship between poor self-monitoring and the presence of alien control symptoms provides support for Frith & Done's account of the origins of these symptoms in schizophrenia.


Fossil evidence for key innovations in the evolution of insect diversity.
Explaining the taxonomic richness of the insects, comprising over half of all described species, is a major challenge in evolutionary biology. Previously, several evolutionary novelties (key innovations) have been posited to contribute to that richness, including the insect bauplan, wings, wing folding and complete metamorphosis, but evidence over their relative importance and modes of action is sparse and equivocal. Here, a new dataset on the first and last occurrences of fossil hexapod (insects and close relatives) families is used to show that basal families of winged insects (Palaeoptera, e.g. dragonflies) show higher origination and extinction rates in the fossil record than basal wingless groups (Apterygota, e.g. silverfish). Origination and extinction rates were maintained at levels similar to Palaeoptera in the more derived Polyneoptera (e.g. cockroaches) and Paraneoptera (e.g. true bugs), but extinction rates subsequently reduced in the very rich group of insects with complete metamorphosis (Holometabola, e.g. beetles). Holometabola show evidence of a recent slow-down in their high net diversification rate, whereas other winged taxa continue to diversify at constant but low rates. These data suggest that wings and complete metamorphosis have had the most effect on family-level insect macroevolution, and point to specific mechanisms by which they have influenced insect diversity through time.


[Oblique muscle dysfunction and cyclotropia].
The disc-fovea angle was calculated from the fundus photo by measuring the distance of fovea to the geometric center of the disc and the distance of fovea to the horizontal meridian through the disc center. The authors found the average disc-fovea angle of 40 normal individuals to be extorsion 7.43 degrees; and the diagnosis of cyclotropia was made for patients with extorsion over 14.20 degrees or intorsion, and those in whom the disc-fovea angle difference between both eyes was 8 degrees or over. In 9 cases of AV syndrome, 86% (12/14) of the V pattern cases associated with inferior oblique overaction were excyclotropic, and 75% (3/4) of the A pattern cases were incyclotropic. In 5 cases of superior oblique paresis, 86% (6/7) of the diseased eyes were excyclotropic, whether the function of the antagonistic was normal. The authors concluded that fundus photos could serve as a basis for diagnosis of cyclotropia.


Identification of myopia-related marker proteins in tilapia retinal, RPE, and choroidal tissue following induced form deprivation.
Experimentally induced myopia is characterized by axial elongation of the eye. The molecular pathways leading to this condition are largely unknown, even though many candidate proteins have been proposed to be involved in this process. This study has identified proteins that were differentially expressed in myopic and control combined retina, retinal pigment epithelium (RPE), and choroidal tissue in tilapia (Oreochromis niloticus). Form deprivation was used to induce myopia in tilapia (n = 3). In this initial study on tilapia retina, RPE and choroid, 2-D differential in gel electrophoresis (DIGE) and mass spectrometry were used to identify differentially expressed proteins. Homology-based gene cloning was used to obtain full sequence data for one of the identified proteins. A total of 18 protein spots separated by 2-D electrophoresis exhibited statistically significant differences in expression between the myopic and contralateral control combined retinal, RPE, and choroidal tissue. Three proteins were identified at a significance level of p < 0.05, as annexin A5 (down-regulated 47%), Gelsolin (down-regulated 27%), and TCP-1 (CCT) (down-regulated 54%). DNA sequencing of tilapia annexin A5 shows an amino acid sequence identity of 84.5% with the homologous Japanese ricefish annexin max2. A proteomics approach has been used to identify differentially expressed proteins in form-deprived combined retinal, RPE, and choroidal tissue from myopic versus normal eyes. The identified proteins may be components of pathways involved in myopia pathogenesis.


Microbiota: dual-faceted player in experimental colitis.
We have recently demonstrated that microbiota stimulation of innate immune pathways is required for T cell spontaneous proliferation and chronic intestinal inflammation. Microbiota promoted spontaneous proliferation of T cells by activating dendritic cells to produce interleukin (IL)-6 via a TLR/Myd88-dependent pathway. Although both CBir1-specific Tcr transgenic (CBir1 Tg) T cells, which are specific for an immunodominant microbiota antigen and OT-II T cells, which are specific for the model antigen ovalbumin, underwent spontaneous proliferation, only CBir1 Tg T cells but not OT-II T cells induced colitis in specific pathogen-free RAG(-/-) mice. Blockade of il-6 or il-6-mediated spontaneous proliferation of CBir1 Tg T cells abrogated colitis induction in this adoptive transfer model. Our data reveal that microbiota serves as a natural adjuvant for T cell spontaneous proliferation and development of chronic intestinal inflammation and that both microbiota stimulation of innate immune cells with subsequent T cell spontaneous proliferation and microbiota antigen activation of antigen-specific Tcr are required for the induction of experimental colitis.


New insights into the roles of NADPH oxidases in sexual development and ascospore germination in Sordaria macrospora.
NADPH oxidase (NOX)-derived reactive oxygen species (ROS) act as signaling determinants that induce different cellular processes. To characterize NOX function during fungal development, we utilized the genetically tractable ascomycete Sordaria macrospora. Genome sequencing of a sterile mutant led us to identify the NADPH oxidase encoding nox1 as a gene required for fruiting body formation, regular hyphal growth, and hyphal fusion. These phenotypes are shared by nor1, lacking the NOX regulator NOR1. Further phenotypic analyses revealed a high correlation between increased ROS production and hyphal fusion deficiencies in nox1 and other sterile mutants. A genome-wide transcriptional profiling analysis of mycelia and isolated protoperithecia from wild type and nox1 revealed that nox1 inactivation affects the expression of genes related to cytoskeleton remodeling, hyphal fusion, metabolism, and mitochondrial respiration. Genetic analysis of nox2, lacking the NADPH oxidase 2 gene, nor1, and transcription factor deletion mutant ste12, revealed a strict melanin-dependent ascospore germination defect, indicating a common genetic pathway for these three genes. We report that gsa3, encoding a G-protein α-subunit, and sac1, encoding cAMP-generating adenylate cyclase, act in a separate pathway during the germination process. The finding that cAMP inhibits ascospore germination in a melanin-dependent manner supports a model in which cAMP inhibits NOX2 activity, thus suggesting a link between both pathways. Our results expand the current knowledge on the role of NOX enzymes in fungal development and provide a frame to define upstream and downstream components of the NOX signaling pathways in fungi.


Hyperactivity and Inattention in Young Patients Born With an Atrial Septal or Ventricular Septal Defect.
Background: Patients with congenital heart defects have a well-established risk of neuropsychiatric comorbidities. Inattention and hyperactivity are three to four times more frequent in children with complex congenital heart defects. We have previously shown a higher burden of overall attention deficit/hyperactivity disorder (ADHD) symptoms in adults with simple congenital heart defects as well. However, it is unknown whether the higher burden of ADHD symptoms is mainly driven by hyperactivity, inattention, or both. Methods: The participants [simple congenital heart defect = 80 (26.6 years old), controls = 36 (25.3 years old)] and a close relative for each (n = 107) responded to the long version of the Conners' Adults ADHD Rating Scales questionnaire. Our primary and secondary outcomes are mean T-scores in the ADHD scores and symptom sub-scores. Results: Patients with simple congenital heart defects reported a higher mean T-score at all three DSM-IV ADHD scores (ADHD-combined: 52.8 vs. 44.9, p = 0.007, ADHD-inattention: 55.5 vs. 46.4, p = 0.002, and ADHD-hyperactivity: 49.4 vs. 44.0, p = 0.03) and in all four ADHD symptom sub-scores (inattention/memory problems: 50.3 vs. 44.2, p = 0.001, hyperactivity/restlessness: 49.7 vs. 45.9, p = 0.03, impulsivity/emotional lability: 50.0 vs. 41.3, p = 0.001, and self-esteem problems: 53.8 vs. 46.3, p = 0.003). The results were maintained after the removal of outliers (incongruent responses), albeit the hyperactivity/restlessness ADHD symptom sub-score lost significance. Self- and informant ratings differed significantly on the ADHD-inattention score for the congenital heart defect group, where informants rated the ADHD-inattention scores better than the congenital heart defect patients rated themselves. Conclusions: Patients with a simple congenital heart defect have a higher symptom burden across all ADHD scores and all symptom sub-scores. The higher burden of ADHD is driven by both inattention and hyperactivity symptoms, though the inattention symptoms seem more prominent. Close relatives were less aware of the inattention symptoms than the congenital heart defect patients themselves. Routine screening for ADHD symptoms may be warranted to facilitate adequate help and guidance as these symptoms are easily overlooked. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT03871881.


Cost-Utility of Attachment-Based Compassion Therapy (ABCT) for Fibromyalgia Compared to Relaxation: A Pilot Randomized Controlled Trial.
A recent study has supported the efficacy of Attachment-Based Compassion Therapy (ABCT) compared to relaxation (REL) for the management of fibromyalgia (FM). The main objective of this paper is to examine the cost-utility of ABCT compared to REL in terms of effects on quality-adjusted life years (QALYs) as well as healthcare costs. Forty-two Spanish patients with FM received 8 weekly group sessions of ABCT or REL. Data collection took place at pre- and 3-month follow-up. Cost-utility of the two treatment groups (ABCT vs. REL) was compared by examining treatment outcomes in terms of QALYs (obtained with the EQ-5D-3L) and healthcare costs (data about service use obtained with the Client Service Receipt Inventory). Data analyses were computed from a completers, ITT, and per protocol approach. Data analysis from the healthcare perspective revealed that those patients receiving ABCT exhibited larger improvements in quality of life than those doing relaxation, while being less costly 3 months after their 8-week treatment program had ended (completers: incremental cost M, 95% CI = €-194.1 (-450.3 to 356.1); incremental effect M, 95% CI = 0.023 QALYs (0.010 to 0.141)). Results were similar using an ITT approach (incremental cost M, 95% CI = €-256.3 (-447.4 to -65.3); incremental effect M, 95% CI = 0.021 QALYs (0.009 to 0.033)). A similar pattern of results were obtained from the per protocol approach. This RCT has contributed to the evidence base of compassion-based interventions and provided useful information about the cost-utility of ABCT for FM patients when compared to relaxation. However, the small sample size and short follow-up period limited the generalizability of the findings.


Smoking habits of parents attending a children's hospital.
This exploratory study surveyed 418 parents attending a paediatric hospital in order to establish their smoking habits and preparedness to quit. We found that 27% were current smokers, of whom 81% were willing to try and quit in the near future. These and the parent's other responses to a questionnaire will be used to develop a programme of smoking cessation for parents attending a children's hospital.


A pharmacy intervention program: recognizing pharmacy's contribution to improving patient care.
An on-line pharmacy intervention program developed to document and evaluate pharmacist's contribution to patient care is described. Over a 1-year period, the number and types of interventions and their impact on patient care were collated and reviewed by a clinical coordinator. Two thousand four hundred ninety-nine interventions were recorded. The most common types of interventions were order clarification/change (18%), pharmacokinetic consult (16%), chart review (13%), restricted drug follow-up (8%), discharge medication screen (7%), initiate drug therapy (6%), drug information (5%), discontinued drug (4%), and therapeutic alternative (4%). There were 3459 impact codes assigned to these interventions. Forty-one percent decreased toxicity, 35% increased efficacy, 17% decreased cost, 16% avoided allergy or drug interaction, 8% improved compliance, and 22% were classified as other. Our analysis found that pharmacy interventions elevated the standard of care and prevented major organ damage and potentially life-threatening events. This program shows that pharmacists play a significant role in improving patient outcomes.


Cytologic findings of collecting duct carcinoma of the kidney.
We report the cytologic features of eight fine-needle aspirations (FNA) and eight exfoliative specimens of collecting duct carcinoma (CDC) obtained from six patients. The four men and two women ranged in age from 27 to 69 years (mean = 45 yr) and all had advanced stage disease at presentation (one stage III, five stage IV). Five of the six patients died of widespread disease, and one is alive and well (mean survival, 28 mo; range, 11-48 mo). The smears of the FNA and exfoliative specimens were scantly to moderately cellular. Tumor cells showed moderate pleomorphism and were arranged primarily in cohesive groups that rarely had a papillary configuration. Nuclei had irregular nuclear contours, coarse chromatin, and one to three nucleoli. In the majority of cases the cytoplasm was finely vacuolated, and occasionally there were large intracytoplasmic vacuoles. Intracytoplasmic mucin was demonstrated in two aspirates. Psammoma bodies were present in four of the seven fluids. In two patients, the cytologic diagnosis was supported by positive immunostaining for high-molecular-weight keratin and Ulex europaeus agglutinin I lectin. Leu M-1 was focally positive in one case and negative in the other. The cytologic features of CDC were readily identified as malignant; however, they were not distinctive and overlapped with those of high-grade renal cell carcinoma with papillary features and transitional cell carcinoma.


¹H-²H cross-relaxation study in a partially deuterated nematic liquid crystal.
A detailed study of the cross-relaxation effects between the ¹H and ²H spins systems is presented in the nematic phase of a 5-cyanobiphenyl (5CB) liquid crystal, partially deuterated at α position (5CB-αd₂). The proton spin-lattice relaxation time was measured at a frequency range from 5 kHz to 100 MHz at a temperature 5 K below the nematic-isotropic phase transition. In the low frequency domain, the spin-lattice relaxation rate (T₁⁻¹) dispersion clearly differs from that of the fully protonated 5CB homologue. At two distinct frequencies, T₁⁻¹ presents two distinct local maxima and for low frequencies T₁⁻¹ presents a stronger frequency dependence when compared with what is observed for 5CB. The T₁⁻¹ dispersion obtained for 5CB-αd₂ for frequencies above 60 kHz was interpreted in terms of the relaxation mechanisms usually accepted to interpret the spin-lattice relaxation in nematic phases in general and 5CB in particular. For lower frequencies it was necessary to consider cross-relaxation contributions between the proton and deuterium reservoirs. A detailed model interpretation of the deuterium quadrupolar dips with respect to the proton-spin relaxation is presented. The analysis of the quadrupolar relaxation independently confirms that the order director fluctuations is the dominant mechanism of proton relaxation in the low frequency domain.


Synthetic atrial peptide inhibits intracellular calcium release in smooth muscle.
The effects of a synthetic atrial peptide (atriopeptin II; AP II) on the agonist-induced intracellular Ca2+ release was examined in the isolated rabbit aorta. The agonist-induced phasic contraction in a Ca2+-free physiological salt solution containing 2 mM ethyleneglycol-bis(beta-aminoethyl-ether)-N,N'-tetraacetic acid (EGTA-PSS) was used as an indicator of the intracellular Ca2+ release. The addition of AP II (10(-9)-10(-7) M) for 15 min to the tissue during the EGTA-PSS exposure caused a dose-dependent inhibition of norepinephrine (NE; 10(-6) M)-induced phasic contraction. The half-maximal inhibiting concentration of AP II was 3 X 10(-9) M, with 10(-7) M AP II causing 91% inhibition. This was confirmed by studying the inhibitory effect of AP II (10(-7) M) on NE-stimulated 45Ca efflux. Furthermore, the internal Ca2+ release by histamine (10(-5) M) and caffeine (25 mM), both of which share this internal Ca2+ pool with NE, was also inhibited by AP II. Thus AP II appears to be a potent inhibitor of the intracellular Ca2+ release that is utilized by various agonists for the activation of vascular smooth muscle. This may be an important mechanism by which AP II produces relaxation of blood vessels.


Estimating the clinical cost of drug development for orphan versus non-orphan drugs.
High orphan drug prices have gained the attention of payers and policy makers. These prices may reflect the need to recoup the cost of drug development from a small patient pool. However, estimates of the cost of orphan drug development are sparse. Using publicly available data, we estimated the differences in trial characteristics and clinical development costs with 100 orphan and 100 non-orphan drugs. We found that the out-of-pocket clinical costs per approved orphan drug to be $166 million and $291 million (2013 USD) per non-orphan drug. The capitalized clinical costs per approved orphan drug and non-orphan drug were estimated to be $291 million and $412 million respectively. When focusing on new molecular entities only, we found that the capitalized clinical cost per approved orphan drug was half that of a non-orphan drug. More discussion is needed to better align on which cost components should be included in research and development costs for pharmaceuticals.


Mechanisms underlying phonotactic steering in the cricket Gryllus bimaculatus revealed with a fast trackball system.
Phonotactic steering behaviour of the cricket G. bimaculatus was analysed with a new highly sensitive trackball system providing a spatial and temporal resolution of 127 microm and 0.3 ms, respectively. Orientation to artificial calling songs started at 45 dB SPL, it increased up to 75 dB SPL and then saturated. When exposed to two identical patterns of different intensity, crickets significantly steered towards the louder sound pattern, whenever the intensity difference was greater than 1 dB. Bilateral latency differences in sound presentation did not always cause clear orientation towards the leading side. The overall walking direction depended on the number of sound pulses perceived from the left or right side with the animals turning towards the side providing the larger number of pulses. The recordings demonstrated rapid changes in walking direction performed even during a chirp. These rapid steering responses occurred with a latency of 55-60 ms, well before the central nervous system had time to evaluate the temporal structure of a whole chirp. When every other sound pulse was presented from opposite directions, the crickets followed the temporal pattern of sound presentation and rapidly steered towards the left and right side. Steering towards individual sound pulses does not agree with the proposal that crickets analyse the quality of sound patterns and then steer towards the better pattern. Rather, these experiments suggest that fast steering to single sound pulses determines the lateral deviation of the animals and that complex auditory orientation emerges from this simple mechanism of auditory steering.


Skull destruction from intracranial metastasis arising from pulmonary squamous cell carcinoma: a case report.
Squamous cell carcinoma of the lung represents 30% of all non-small cell lung carcinomas. It arises from dysplasia of squamous epithelium of the bronchi and is strongly associated with cigarette smoking. Squamous cell carcinoma of the lung is known to produce metastases in the brain parenchyma. We present the case of an 80-year-old indigenous Australian man with an unusual presentation of metastatic carcinoma of the lung. The case demonstrated a squamous cell carcinoma of the lung with an intracranial metastatic lesion destroying the parietal bone and extending into the extracranial soft tissue. A visible deformity as a result of the metastasis was evident on physical examination and computed tomography demonstrated extensive bone destruction. The authors were unable to find a case of this occurring from a squamous cell carcinoma of the lung anywhere in the world literature. The case report demonstrates an unusual disease presentation with a rare intracranial metastasis invading through the skull.


Why youth smoke? An exploratory community-based study from Chandigarh Union Territory of northern India.
Tobacco use is a serious public health challenge in several regions of the world, including India. Increasingly, steps are being taken at policy level to curb the problem. This study was done to find out the determinants of tobacco use so that effective intervention programs can be designed and implemented for the prevention and cessation of this growing pandemic. A community-based cross-sectional study was done adapting Global Youth Tobacco Survey questionnaire prepared by the Centre for Disease Control, Atlanta, among youth (15-24 years). Patterns of smoking and their determinants were calculated using univariate and multivariate analyses. Prevalence of current smoking among youth was 20.4% (95% confidence interval: 16.9-23.9%). Male sex, smoking peers, cigarette advertisements, and feeling comfortable in social gatherings were significant determinants for smoking after adjusting for all explanatory variables. Strict enforcement of regulations pertaining to cigarette advertisements in any form, enabling environment and community interventions focusing on parents and peers are required for effective control of tobacco problem among youth in India.


Disruption of cell death signaling in cancer: impact on disease prognosis and response to therapy.
Disruption of cell cycle and apoptosis signaling pathways are key events during tumorigenesis, tumor progression and development of resistance against anticancer therapies. Thus, the analysis of functional alterations within these signaling cascades is of utmost importance for the understanding of resistance mechanisms, clinical outcome and risk-adapted treatment strategies. Key signaling pathways involved in the treatment resistance include the p53/p14ARF signaling complex and the mitochondrial apoptosis machinery. Apart from the direct genetic events, these signaling cascades are subject to epigenetic modulations implied by the tumor microenvironment.


Phosphorylation of CENP-A on serine 7 does not control centromere function.
CENP-A is the histone H3 variant necessary to specify the location of all eukaryotic centromeres via its CENP-A targeting domain and either one of its terminal regions. In humans, several post-translational modifications occur on CENP-A, but their role in centromere function remains controversial. One of these modifications of CENP-A, phosphorylation on serine 7, has been proposed to control centromere assembly and function. Here, using gene targeting at both endogenous CENP-A alleles and gene replacement in human cells, we demonstrate that a CENP-A variant that cannot be phosphorylated at serine 7 maintains correct CENP-C recruitment, faithful chromosome segregation and long-term cell viability. Thus, we conclude that phosphorylation of CENP-A on serine 7 is dispensable to maintain correct centromere dynamics and function.


[Blood groups and discoid lupus erythematosus].
Discoid lesion is the commonest cutaneous finding in lupus erythematosus and chronic types have their own immunological features, with Th1 inflammation profile. Although many diseases have association with blood-group systems, this fact was not enlightened in discoid lupus erythematosus. To investigate the association between blood groups (ABO and Rh) and discoid lupus erythematosus. A prospective cross-sectional study assessing clinical information, blood group systems (ABO and Rh), FAN and C4 serum levels from discoid lupus patients without characteristics of systemic disease, was carried out at a clinic from a Brazilian university hospital. Sixty-nine patients were enrolled in the study, 71.0% were females (p1:160 in 31.9%, and low levels of C4 in 8.7%. There was no significant difference between the frequency of blood groups from discoid lupus patients and local population, however, blood group A was associated to disseminate forms of the disease (OR 4.1 and p < 0.05). Discoid lupus erythematosus patients with ABO and Rh blood groups exhibit similar frequencies as in the general population; nevertheless, disseminate clinical forms were more prevalent among group A patients.


Molecular epidemiology of Mycobacterium abscessus infections in a subtropical chronic ventilatory setting.
The aim of this study was to investigate the high level of pulmonary Mycobacterium abscessus infections and implement a surveillance programme among 43 ventilator-dependent patients, 15 with pulmonary M. abscessus infections, in a hospital long-term respiratory care ward (RCW) in central Taiwan. M. abscessus isolates were obtained from 35 patients in the RCW of hospital A, 6 patients in the RCWs of another three hospitals (B, C and D), and from 4 water sources in two of the hospitals (A and B). Strains were characterized by methods including hsp65 PCR-RFLP and PFGE. The patients were followed-up by chest X-ray for 1 year. All clinical isolates were type I and II, and belonged to ten distinct clusters of PFGE patterns. Five clinical strains in two hospitals belonged to a single cluster, whilst four clinical strains in the other two hospitals belonged to a single unique cluster. The strains from hospital A fell into nine clusters and were distinct from the strains isolated from the water supply. Patients infected with type I strains showed a significantly more rapid progression of disease. The number of different strains involved suggested either that there had been a polyclonal outbreak or that a high level of endemic infections was present in the RCW of hospital A. This and the lack of homology between the clinical and environmental isolates from hospital A raised the possibility that pulmonary M. abscessus infections may have been spread by the movement of patients between RCWs, a routine practice in Taiwan's integrated delivery system.


Brain changes underlying cognitive dysfunction in diabetes: what can we learn from MRI?
Diabetes is associated with cognitive dysfunction and an increased risk of dementia. This article addresses findings with brain MRI that may underlie cognitive dysfunction in diabetes. Studies in adults with type 1 diabetes show regional reductions in brain volume. In those with a diabetes onset in childhood, these volume reductions are likely to reflect the sum of changes that occur during brain development and changes that occur later in life due to exposure to diabetes-related factors. Type 2 diabetes is associated with global brain atrophy and an increased burden of small-vessel disease. These brain changes occur in the context of aging and often also in relation to an adverse vascular risk factor profile. Advanced imaging techniques detect microstructural lesions in the cerebral gray and white matter of patients with diabetes that affect structural and functional connectivity. Challenges are to further unravel the etiology of these cerebral complications by integrating findings from different imaging modalities and detailed clinical phenotyping and by linking structural MRI abnormalities to histology. A better understanding of the underlying mechanisms is necessary to establish interventions that will improve long-term cognitive outcomes for patients with type 1 and type 2 diabetes.


[Endoscopic papillotomy in patients with post-cholecystectomy syndrome].
In benign stenosis of the major duodenal papilla, choledocholithiasis and their combination, the performance of endoscopic papillotomy permitted in 89.4% of the 113 patients to avoid the repeated operative intervention. Complications developed in 5.3% of patients, the postoperative lethality was 0.8%.


The rise and fall of red cell mass measurement in polycythemia vera.
The total blood volume (BV) consists of the part occupied by red blood cells (RBC), which is referred to as red cell mass (RCM), and that occupied by plasma (ie, plasma volume). Quantitative laboratory measurements that are pertinent to RBC, including RBC count, hematocrit (Hct), and hemoglobin (Hgb) are expressed in reference to a given volume of whole blood and are therefore influenced by plasma volume. Consequently, a "direct" RCM measurement has been promoted as a more accurate indicator of the body's red cell content. In accordance with this view, an international group of investigators, then identified as the Polycythemia Vera Study Group (PVSG), recommended that RCM be measured and only if elevated should a patient be considered for participation in a series of clinical trials in PV that were conducted more than 30 years ago. By default, the 'study eligibility' criteria used in these studies became 'diagnostic' criteria without any systematic evaluation for diagnostic accuracy. Furthermore, over the years, it has become evident that RCM measurement is a cumbersome and costly test that is also suboptimal in its diagnostic accuracy. As a result, the specific procedure has been abandoned by the majority of hematologists in certain countries and instead physicians are increasingly using bone marrow histology, serum erythropoietin level, and other disease-characteristic biologic markers as diagnostic tools.


Preoperative Bone Mineral Density and Bone Turnover in Women Before Primary Knee Arthroplasty.
The preoperative prevalence of osteoporosis and/or osteopenia and overall bone quality in prospective total knee arthroplasty (TKA) patients may affect the postoperative outcome after prosthetic insertion into the bone. The purpose of this study is to determine the baseline bone mineral density (BMD) and bone turnover in preoperative, female, primary TKA patients. We prospectively measured the lumbar spine and hip BMDs using dual-energy X-ray absorptiometry (DEXA) scans in a cohort of 119 knees (107 patients) one day before surgery. We also assessed bone turnover using urinary levels of N-telopeptide (NTX), a type I collagen crosslinker, normalized to creatinine. The prevalence of osteoporosis by DEXA scan (T-score ≤ -2.5) among the TKAs was 12% in the spine and 10% in the hip. Eighty-three knees (70%) had osteopenia or osteoporosis of either the spine or hip. The mean T-score of the spine was -0.7 (SD 1.6), which is within normal limits, and of the hip was -1.2 (SD 1.0), which is defined as osteopenia. The mean Z-scores of 0.9 (SD 1.4) in the spine and 0.6 (SD 0.9) in the hip were positive. The median urinary NTX/creatinine ratio was elevated at 58.1 (interquartile range: 13.7 to 188.4). Based on Z-scores, the TKA patients had higher spine and hip BMDs than the age-matched general population. Elevated NTX levels may suggest a systemic or local abnormal bone turnover. Further study is needed to determine whether such turnover, as a type of patient-related medical systemic disorder, affects postoperative clinical outcomes.


Cenicriviroc for the treatment of non-alcoholic steatohepatitis and liver fibrosis.
Nonalcoholic fatty liver disease (NAFLD) has an increasing prevalence worldwide. At present, no specific pharmacotherapy is approved for NAFLD. Simple steatosis and nonalcoholic steatohepatitis (NASH) can progress to liver fibrosis that is associated with mortality in NAFLD. The recruitment of inflammatory monocytes and macrophages via chemokine receptor CCR2 as well as of lymphocytes and hepatic stellate cells via CCR5 promote the progression of NASH to fibrosis. Areas covered: I summarize preclinical and clinical data on the efficacy and safety of the dual CCR2/CCR5 inhibitor cenicriviroc (CVC, also TBR-652 or TAK-652) for the treatment of NASH and fibrosis. In animal models of liver diseases, CVC potently inhibits macrophage accumulation in the liver and ameliorates fibrosis. In a phase 2b clinical trial (CENTAUR) on 289 patients with NASH and fibrosis, CVC consistently demonstrated liver fibrosis improvement after 1 year of therapy and had an excellent safety profile, leading to the implementation of a phase 3 trial (AURORA). Expert opinion: Preclinical and clinical data support the development of CVC as a safe and potent antifibrotic agent. However, open questions around CVC are the durability of antifibrotic responses, divergent effects on NASH versus fibrosis, potential long-term concerns and the expected path to approval.


A biomarker-responsive T2ex MRI contrast agent.
This study investigated a fundamentally new type of responsive MRI contrast agent for molecular imaging that alters T2 exchange (T2ex ) properties after interacting with a molecular biomarker. The contrast agent Tm-DO3A-oAA was treated with nitric oxide (NO) and O2 . The R1 and R2 relaxation rates of the reactant and product were measured with respect to concentration, temperature, and pH. Chemical exchange saturation transfer (CEST) spectra of the reactant and product were acquired using a 7 Tesla (T) MRI scanner and analyzed to estimate the chemical exchange rates and r2ex relaxivities. The reaction of Tm-DO3A-oAA with NO and O2 caused a 6.4-fold increase in the r2 relaxivity of the agent, whereas r1 relaxivity remained unchanged, which demonstrated that Tm-DO3A-oAA is a responsive T2ex agent. The effects of pH and temperature on the r2 relaxivities of the reactant and product supported the conclusion that the product's benzimidazole ligand caused the agent to have a fast chemical exchange rate relative to the slow exchange rate of the reactant's ortho-aminoanilide ligand. T2ex MRI contrast agents are a new type of responsive agent that have good detection sensitivity and specificity for detecting a biomarker, which can serve as a new tool for molecular imaging. Magn Reson Med 77:1665-1670, 2017. © 2016 International Society for Magnetic Resonance in Medicine.


Distribution of Different PML/RARα bcr Isoforms in Indian Acute Promyelocytic Leukemia (APL) Patients and Clinicohematological Correlation.
Acute promyelocytic leukemia (APL), an AML subtype, is characterized morphologically by abnormal promyelocytes. Molecular studies show three possible bcr isoforms of PML-RARα fusion gene. This study undertakes analysis of PML-RARα bcr isoforms and their correlation with haematological parameters and response to treatment in Indian patients. To study different PML-RARα bcr isoforms in Indian patients and to find any correlation with various haematological parameters and response to treatment. Patients diagnosed as APL on morphology or flowcytometry and confirmed by RQ PCR were included in the study. Treated APL patients or patients with relapse and on follow-up were excluded from the study. Twenty patients over thirty one months period were included. The clinical, haematological & morphological features were analysed, the latter using routine & special cytochemical stains on blood and bone marrow. Flow cytometric evaluation using 4-color Beckman Coulter FC 500 and molecular studies using RT PCR Fusion Quant® kits for bcr-1, bcr-2 and bcr-3 of PML-RARα bcr isoforms on the instrument Rotor Gene™ 3000 were performed. Student t test was applied to correlate different bcr isoforms with various haematological parameters and response to treatment. In our study, M:F ratio was 1.5:1 with median age 42 years, Hb - 8.0 g/dl, TLC-7900/μl, and platelet - 35000/μl and varied clinical presentation. Four patients were microgranular variants, and the rest were hypergranular. MPO and CAE positivity were100% and for NSE it was 33.33%. Molecular analysis revealed PML-RARα isoforms of bcr1 in 42.85%, bcr2 in 14.28% and bcr3 in 38.09% patients. No correlation was found between PML-RARα bcr isoforms, different haematological parameters and response to treatment. Higher incidence of PML-RARα bcr-1 isoform was found in Indian APL patients with no significant correlation between different haematological parameters and response to treatment.


Toxicological aspects of aluminum poisoning in clinical nephrology.
Several dialysis-related diseases can be ascribed to an increased body burden of aluminum (Al). Determination of this metal in clinical samples is of great assistance in the diagnosis, treatment and prevention of Al intoxication. As there are many pitfalls in the estimation of Al in biological fluids, the performance of these assays should be restricted to laboratories specializing in the human toxicology of metals, which are experienced in flameless atomic absorption spectrophotometry and which participate in an external quality assessment scheme. One laboratory can serve ten dialysis centers. Serum, plasma or blood are equivalent samples for the biological monitoring of Al. The assay should be performed once in a three month period, but when oral Al-containing medication is changed, toxic symptoms are suspected, or levels exceeding 100 micrograms/l are observed, Al should be determined much more frequently, e.g., once weekly. A high incidence of symptoms associated with Al poisoning is seen in patients with serum concentrations (AlS) greater than 100 micrograms/l, and it is therefore recommended to keep AlS well below this level. Scalp hair appears to be of no use for the assessment of the body burden of Al in the individual patient. For "environmental monitoring", regular analysis of the dialysis fluid is necessary. The concentration of Al in this fluid should not exceed 5 micrograms/l; higher levels lead to a positive Al balance during dialysis and may eventually result in poisoning.


Mitochondria-targeting sequence, a multi-role sorting sequence recognized at all steps of protein import into mitochondria.
The intracellular sorting of newly synthesized precursor proteins (preproteins) to mitochondria depends on the "mitochondria-targeting sequence" (MTS), which is located at the amino termini of the preproteins. MTS is required, however, not only for targeting newly synthesized preproteins to mitochondria, but also for all the following steps along the mitochondrial protein import pathway. MTS of nascent preproteins is first recognized by a cytoplasmic molecular chaperone, MSF, and then by Tom70 and Tom20 of the mitochondrial outer membrane receptor complex, Tom5 and Tom40 of the outer membrane protein translocation machinery, Tim23 of the inner membrane protein translocation machinery, and finally the processing peptidase, MPP, in the matrix. MTS is a multi-role sorting sequence which specifically interacts with various components along the mitochondrial protein import pathway. Recognition of MTS at multiple steps during the import of preproteins may contribute to the strict sorting of proteins destined for mitochondria.


Malnutrition: more than the eye can see.
To assess the magnitude of malnutrition in a hospital setting and to relate anthropometric measures to the clinical diagnosis of malnutrition. A descriptive study whereby anthropometric measures (length and weight) were taken of every child under the age of five years who was admitted to the hospital. The anthropometric data were analysed using the EPI-Info statistical package, which calculates z-scores for weight-for-age, weight-for-height and height-for-age. As reference curve, the reference growth curves of the NCHS were used. Of all the children who were classified as being malnourished, it was recorded if the clinical diagnosis of malnutrition was made at the time of admission or during the hospital stay. Misikhu Mission Hospital, western Kenya. Every child under the age of five years who was admitted to the hospital, was eligible to enter the study. The data of 1130 children were used. The data of 40 other children who were admitted in this period were not complete and could therefore not be used. An overall percentage for malnutrition of 44.3 was found. Only fourteen per cent of the malnourished children were clinically diagnosed as having malnutrition. Anthropometric measures are an easy, but time-consuming way of identifying children with malnutrition, it identifies more children with malnutrition than clinical diagnosis alone. Therefore it should be considered to implement standardised anthropometry in a hospital setting.