Structure and function of Rubisco.
Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is the major enzyme assimilating CO(2) into the biosphere. At the same time Rubisco is an extremely inefficient catalyst and its carboxylase activity is compromised by an opposing oxygenase activity involving atmospheric O(2). The shortcomings of Rubisco have implications for crop yield, nitrogen and water usage, and for the global carbon cycle. Numerous high-resolution crystal structures of different forms of Rubisco are now available, including structures of mutant enzymes. This review uses the information provided in these structures in a structure-based sequence alignment and discusses Rubisco function in the context of structural variations at all levels--amino acid sequence, fold, tertiary and quaternary structure--with an evolutionary perspective and an emphasis on the structural features of the enzyme that may determine its function as a carboxylase.


[Imaginaries present in the training of health professionals in Brazil and the horizons of supplementary care regulation].
This paper is part of a research into the imaginaries present in the training of health professionals with respect to the regulation and exercise of their profession. The professions selected were medicine, odontology and psychology, for which emerging courses require evaluation by the National Health Council. The imaginaries were understood as operators of the virtual and the real with the potential of affirming or denying forms and contents in relation to being a professional or being in the profession. There was evidence for an imaginary of free exercise of the profession in the public sector on state level, where more experience with diseases and diversities of suffering would be gained. The ideal work place would be the private sector allowing for free choices for both professionals and users, but not without connection to the public sector for providing experience, study opportunities and the possibility of research scholarships and overseas training. Despite the expectations in relation to the private sector, currently there is no education in the field of supplementary care or about the meanings of the regulation proposed by the Brazilian Health System.


CYR61 stimulates human skin fibroblast migration through Integrin alpha vbeta 5 and enhances mitogenesis through integrin alpha vbeta 3, independent of its carboxyl-terminal domain.
CYR61, an angiogenic factor and a member of the CCN protein family, is an extracellular matrix-associated, heparin-binding protein that mediates cell adhesion, promotes cell migration, and enhances growth factor-stimulated cell proliferation. CYR61 induces angiogenesis and promotes tumor growth in vivo and is expressed in dermal fibroblasts during cutaneous wound healing. It has been demonstrated recently that adhesion of primary skin fibroblasts to CYR61 is mediated through integrin alpha(6)beta(1) and cell surface heparan sulfate proteoglycans, resulting in adhesive signaling and up-regulation of matrix metalloproteinases 1 and 3. CYR61 is composed of four discrete structural domains that bear sequence similarities to the insulin-like growth factor-binding proteins, von Willebrand factor type C repeat, thrombospondin type 1 repeat, and a carboxyl-terminal (CT) domain that resembles cysteine knots found in some growth factors. In this study, we show that a CYR61 mutant (CYR61DeltaCT) that has the CT domain deleted is unable to support adhesion of primary human skin fibroblasts but is still able to stimulate chemotaxis and enhance basic fibroblast growth factor-induced mitogenesis similar to wild type. In addition, fibroblast migration to CYR61 is mediated through integrin alpha(v)beta(5) but not integrins alpha(6)beta(1) or alpha(v)beta(3). Furthermore, we show that CYR61 binds directly to purified integrin alpha(v)beta(5) in vitro. By contrast, CYR61 enhancement of basic fibroblast growth factor-induced DNA synthesis is mediated through integrin alpha(v)beta(3), a known receptor for CYR61 that mediates CYR61-dependent cell adhesion and chemotaxis in vascular endothelial cells. Thus, CYR61 promotes primary human fibroblast adhesion, migration, and mitogenesis through integrins alpha(6)beta(1), alpha(v)beta(5), and alpha(v)beta(3), respectively. Together, these findings establish CYR61 as a novel ligand for integrin alpha(v)beta(5) and show that CYR61 interacts with distinct integrins to mediate disparate activities in a cell type-specific manner.


Rapid optimisation of cellulolytic enzymes ratios in Saccharomyces cerevisiae using in vitro SCRaMbLE.
For the economic production of biofuels and other valuable products from lignocellulosic waste material, a consolidated bioprocessing (CBP) organism is required. With efficient fermentation capability and attractive industrial qualities, Saccharomyces cerevisiae is a preferred candidate and has been engineered to produce enzymes that hydrolyze cellulosic biomass. Efficient cellulose hydrolysis requires the synergistic action of several enzymes, with the optimum combined activity ratio dependent on the composition of the substrate. In vitro SCRaMbLE generated a library of plasmids containing different ratios of a β-glucosidase gene (CEL3A) from Saccharomycopsis fibuligera and an endoglucanase gene (CEL5A) from Trichoderma reesei. S. cerevisiae, transformed with the plasmid library, displayed a range of individual enzyme activities and synergistic capabilities. Furthermore, we show for the first time that 4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-D-cellopentaoside (BPNPG5) is a suitable substrate to determine synergistic Cel3A and Cel5A action and an accurate predictive model for this synergistic action was devised. Strains with highest BPNPG5 activity had an average CEL3A and CEL5A gene cassette copy number of 1.3 ± 0.6 and 0.8 ± 0.2, respectively (ratio of 1.6:1). Here, we describe a synthetic biology approach to rapidly optimise gene copy numbers to achieve efficient synergistic substrate hydrolysis. This study demonstrates how in vitro SCRaMbLE can be applied to rapidly combine gene constructs in various ratios to allow screening of synergistic enzyme activities for efficient substrate hydrolysis.


Effects of adjunct galantamine to risperidone, or haloperidol, in animal models of antipsychotic activity and extrapyramidal side-effect liability: involvement of the cholinergic muscarinic receptor.
The acetylcholine esterase inhibitor/cholinergic nicotinic receptor (nAChR) allosteric modulator galantamine (Gal) is used against cognitive impairment in Alzheimer's disease. Negative/cognitive and psychotic symptom improvement in schizophrenia by adjunct Gal to antipsychotic drugs (APDs) has been reported. Cognitive symptoms in schizophrenia may involve brain prefrontal hypo-dopaminergia. Experimental data by others indicate nAChR involvement in animal pro-cognitive effects of Gal. The role of nAChRs in antipsychotic effects by Gal has, however, not been elucidated. Using the conditioned avoidance response (CAR) and the catalepsy tests for antipsychotic activity and extrapyramidal side-effect (EPS) liability, respectively, we here investigated the effects of adjunct Gal (1.25 mg/kg) to the typical APD haloperidol (Hal) (0.05 mg/kg), or the atypical APD risperidone (Ris) (0.2 mg/kg), in rats. Adjunct Gal significantly enhanced APD-like effects by low doses of Hal or Ris, but showed a safe EPS liability profile only in combination with Ris. Pretreatment with the muscarinic receptor (mAChR) antagonist scopolamine, but not the nAChR antagonist mecamylamine, completely reversed the enhancing effects of adjunct Gal to Hal treatment, in the CAR test. While the nAChR-modulating properties of Gal probably contribute to pro-cognitive activity, as shown by others, the present data suggest that any contribution to antipsychotic activity by Gal is mediated primarily via mAChRs. This property combination of Gal may offer a unique, favourable therapeutic profile for schizophrenia treatment.


Amyloid- and tau-PET imaging in a familial prion kindred.
To study the in vivo binding properties of 18F-AV-1451 (tau-PET) and Pittsburgh compound B (PiB-PET) in a unique kindred with a familial prion disorder known to produce amyloid plaques composed of prion protein alongside Alzheimer disease (AD)-like tau tangles. A case series of 4 symptomatic family members with the 12-octapeptide repeat insertion in the PRNP gene were imaged with 3T MRI, PiB-PET, and tau-PET in their fourth decade of life. There was significant neocortical uptake of the tau-PET tracer in all 4 familial prion cases. However, PiB-PET images did not demonstrate abnormally elevated signal in neocortical or cerebellar regions for any of the patients. In vivo detection of molecular hallmarks of neurodegenerative diseases will be a prerequisite to well-conducted therapeutic trials. Understanding the in vivo behavior of these PET biomarkers in the setting of various neurodegenerative processes is imperative to their proper use in such trials and for research studies focused on the basic neurobiology of neurodegeneration. This study supports the high specificity of neocortical 18F-AV-1451 binding to AD-like tau and the lack of PiB binding to PrP plaques. It is uncertain how early in the disease course tau pathology appears in the brains of individuals who carry this PRNP gene mutation or how it evolves throughout the disease course, but future longitudinal 18F-AV-1451 imaging of symptomatic and asymptomatic individuals in this kindred will help address these uncertainties.


Cancer incidence among an HIV-infected cohort. Pulmonary Complications of HIV Infection Study Group.
Malignancies, particularly Kaposi's sarcoma and non-Hodgkin's lymphoma (NHL), are associated with human immunodeficiency virus (HIV) infection. Cancer incidence among 1,073 asymptomatic HIV-infected individuals from the Pulmonary Complications of HIV Infection Study cohort, persons from six states followed from 1988 to 1994, was examined. Total cancer incidence was 3.99/100 person-years; for Kaposi's sarcoma, incidence was 2.64 cases/100 person-years, and for NHL, it was 1.18 cases/100 person-years. Total cancer (n = 156 cases) was higher among nonblacks than among blacks (rate ratio = 2.8, 95% confidence interval 1.3-6.1), with similar results for Kaposi's sarcoma and NHL. The rate of lung cancer (n = 5) among white, homosexual/bisexual males was 0.18 per 100 person-years, suggesting a high risk of lung cancer.


Landscape epidemiology of plant diseases.
Many agricultural landscapes are characterized by a high degree of heterogeneity and fragmentation. Landscape ecology focuses on the influence of habitat heterogeneity in space and time on ecological processes. Landscape epidemiology aims at applying concepts and approaches originating from landscape ecology to the study of pathogen dynamics at the landscape scale. However, despite the strong influence that the landscape properties may have on the spread of plant diseases, landscape epidemiology has still received little attention from plant pathologists. Some recent methodological and technological progress provides new and powerful tools to describe and analyse the spatial patterns of host-pathogen interactions. Here, we review some important topics in plant pathology that may benefit from a landscape perspective. These include the influence of: landscape composition on the global inoculum pressure; landscape heterogeneity on pathogen dynamics; landscape structure on pathogen dispersal; and landscape properties on the emergence of pathogens and on their evolution.


Population differences of two coding SNPs in pigmentation-related genes SLC24A5 and SLC45A2.
The two genes SLC24A5 and SLC45A2 were recently identified as major determinants of pigmentation in humans and in other vertebrates. The allele p.A111T in the former gene and the allele p.L374F in the latter gene are both nearly fixed in light-skinned Europeans, and can therefore be considered ancestry informative marker (AIMs). AIMs are becoming useful for forensic identification of the phenotype from a DNA profile sampled, for example, from a crime scene. Here, we generate new allelic data for these two genes from samples of Chinese, Uygurs, Ghanaians, South African Xhosa, South African Europeans, and Sri Lankans (Tamils and Sinhalese). Our data confirm the earlier results and furthermore demonstrate that the SLC45A2 allele is a more specific AIM than the SLC24A5 allele because the former clearly distinguishes the Sri Lankans from the Europeans.


Myocardial protective effect of captopril cardioplegia on ischemia and reperfusion injury in the isolated rabbit heart.
The myocardial protective effect of captopril cardioplegia was investigated through the recovery of cardiac functions, biochemical changes and ultrastructural assays in the isolated working rabbit heart after hypothermic ischemic arrest for 3 hours. Hearts were randomly divided into 3 groups with 8 in each. Another 3 groups of hearts (8 in each) were arranged for studying the biochemical changes and ultrastructure during the ischemic period. In the control group (I), the hearts were protected by the modified St. Thomas' cardioplegic solution No. I, while in Group II and III the cardioplegic solutions contained 2.3 mumol/L and 23 mumol/L captopril respectively. The results showed that the percent recoveries of cardiac function in the 23 mumol/L captopril group compared with the control group were 97.9 +/- 9.8% vs 84.9 +/- 7.3% in left ventricular peak systolic pressure (P less than 0.01), 118 +/- 33% vs 79 +/- 20% in +dp/dt max (P less than 0.01), 111 +/- 22% vs 75 +/- 18% in -dp/dt max (P less than 0.01), 145.6 +/- 44.8% vs 76.7 +/- 18% in coronary flow (P less than 0.01), 120.2 +/- 45.7% vs 36.4 +/- 22.2% in aortic flow (P less than 0.01), 127.4 +/- 27.3% vs 56.1 +/- 14% in cardiac output (P less than 0.01), and 118.2 +/- 33.5% vs 52.5 +/- 19.9% in stroke volume (P less than 0.01) respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Distinctive localization and opposed roles of vasohibin-1 and vasohibin-2 in the regulation of angiogenesis.
We recently isolated a novel angiogenesis inhibitor, vasohibin-1, and its homologue, vasohibin-2. In this study we characterize the role of these 2 molecules in the regulation of angiogenesis. In a mouse model of subcutaneous angiogenesis, the expression of endogenous vasohibin-1 was low in proliferating ECs at the sprouting front but high in nonproliferating endothelial cells (ECs) in the termination zone. In contrast, endogenous vasohibin-2 was preferentially expressed in mononuclear cells mobilized from bone marrow that infiltrated the sprouting front. When applied exogenously, vasohibin-1 inhibited angiogenesis at the sprouting front where endogenous vasohibin-1 was scarce but did not influence vascularity in the termination zone where endogenous vasohibin-1 was enriched. Exogenous vasohibin-2 prevented the termination of angiogenesis in the termination zone and increased vascularity in this region. Angiogenesis was persistent in the termination zone in the vasohibin-1 knockout mice, whereas angiogenesis was deficient at the sprouting front in the vasohibin-2 knockout mice. Supplementation of deficient proteins normalized the abnormal patterns of angiogenesis in the vasohibin knockout mice. These results indicate that vasohibin-1 is expressed in ECs in the termination zone to halt angiogenesis, whereas vasohibin-2 is expressed in infiltrating mononuclear cells in the sprouting front to promote angiogenesis.


Effect of antibiotics and sedatives on normal neutrophil nicotinamide-adenine dinucleotide phosphate-reduced oxidase activity.
The effects of antibiotics and other commonly used medications on the human polymorphonuclear neutrophil leukocytes' (PMNs) nicotinamide-adenine dinucleotide phosphate-reduced (NADPH) oxidase activity have been investigated in vitro. Five antibiotics (penicillin G sodium, cefamandole nafate, metronidazole hydrochloride, clindamycin phosphate, and tobramycin sulfate, and a triple combination of penicillin G sodium-metronidazole hydrochloride-tobramycin sulfate) and two sedatives (morphine sulfate and diazepam) were incubated with normal human PMNs at therapeutic, infratherapeutic, and supratherapeutic drug levels. The superoxide dismutase-inhibitable, NADPH-dependent reduction of cytochrome C in the PMNs was studied after stimulation with formyl-methionyl-leucine-phenylalanine. Tobramycin sulfate and the triple combination of penicillin G sodium-metronidazole hydrochloride-tobramycin sulfate significantly reduced the NADPH oxidase activity at all dosages studied. Clindamycin phosphate, morphine sulfate, and diazepam also showed significant reduction at therapeutic and supratherapeutic concentrations. Penicillin G sodium, cefamandole nafate, and metronidazole hydrochloride did not cause a decrease in enzyme activity at any levels tested. We conclude that NADPH oxidase activity can be adversely affected by the circulating levels of common antibiotics and sedatives.


Microbiology Education in Nigeria: Common Training Deficits Could Be Addressed with Modern Educational Tools†.
Microbiology has developed as an important field of scientific training in Nigeria over the years. It is one of the early scientific fields introduced to the country. It has contributed to the development of the scientific identity of the country, through training of personnel and applications aimed at addressing microbial diseases that present a public health concern. Recent diseases, especially of microbial origin, have however exposed the poor state of microbiology in Nigeria. Recurrent but avoidable infectious diseases such as diarrhea and cholera have led to the loss of several lives. Although the situation is complex, at least some of these shortcomings could be compensated for by enhancing the quality of training delivered to future microbiologists and related healthcare professionals and educators. An analysis of microbiology curricula in Nigerian institutions compared with those of developed countries identified several gaps. Although inadequacy in training resources, expert faculty, and funding are well recognized, it is believed that several of these training gaps could be addressed through innovative educational approaches. In this paper, I discuss how key microbiology education weaknesses in the Nigerian context could be overcome using modern, low-cost educational tools that are accessible even in the face of existing institutional challenges. Topics such as molecular cloning and PCR are discussed, together with how specific virtual classroom and related internet-based tools can be applied to teach these topics in resource-limited contexts. This paper should therefore be relevant to microbiology educators in most developing countries.


An in vitro study examining a novel suction curette device for lumbar discectomy compared with standard manual discectomy.
OBJECTIVE This purpose of this study was to objectively evaluate and assess the efficacy and efficiency of discectomy and endplate preparation during transforaminal lumbar interbody fusion (TLIF) using traditional manual instrumentation versus a novel suction discectomy curette. Transforaminal lumbar interbody fusion is the most widely used approach for lumbar arthrodesis, and its success depends on the ability to achieve fusion. Complete preparation of intervertebral disc space (removal of the nucleus, endplate cartilage, and margin of inner annulus) is the surgical goal. Performing an adequate discectomy requires numerous instrument passes, increasing surgical time and the risk of complications. METHODS Four experienced spinal surgeons performed transforaminal discectomies from T-12 to S-1 on 5 whole-body cadavers. Each level (n = 26) was randomly assigned to either a control group using traditional instruments (12 levels) or to a suction curette group (14 levels). The time required to perform the discectomy and the number of passes through the annulus were recorded. Motion segments were dissected and analyzed by digital photogrammetric analysis. The intervertebral disc and the discectomy cross-sectional areas were measured on both superior and inferior images of each dissected surgical level. Areas were divided into 4 quadrants based on a midsagittal and midcoronal axis and analyzed for regional efficiency. In addition, a cross-sectional area of bony endplate (the area still covered with cartilage) and an area of endplate perforation were evaluated. RESULTS There was no significant difference in surgical time between the techniques (7:51 ± 2:43 minutes in the manual discectomy [MD] group and 7:06 ± 3:33 minutes in the suction curette discectomy [SD] group). There were significantly fewer (p < 0.01) instrument passes in the SD group (13 passes) compared with the MD group (43 passes). For both techniques, the amount of disc removed depended upon the anatomical region, with the posterior-contralateral side having the least amount of disc material removed. There was significantly less (p < 0.01) disc material removed in the MD group (38%) compared with the SD group (48%). The amount of disc material removed was significantly more (p < 0.05) in each quadrant when comparing the SD and MD groups, with the anterior regions showing the largest difference. For both techniques, the preparation of the endplate within the discectomy area resulted in a mostly cartilaginous interface (50% MD, 48% SD); a smaller amount of bony interface area (31% MD, 38% SD); and a smaller amount of perforation to the interface area (19% MD, 13% SD). There were no significant differences between the groups in terms of endplate preparation. CONCLUSIONS The improved discectomy observed with the suction curette device could potentially improve the clinical fusion rate.


Biological therapy in polymyalgia rheumatica.
Polymyalgia rheumatica (PMR) is an inflammatory rheumatic disease of the elderly, treated mainly with systemic corticosteroids. The frequency of side effects of steroids is high in this aged population and increased due to comorbidities. The use of biological treatments could be of interest in this condition. This review takes into account literature data from the PubMed and clinical trial databases concerning the results of the use of biological treatments in PMR, in terms of efficacy and safety of these treatments. Current data do not allow us to identify any particular efficacy of the various anti-TNF agents used in the treatment of PMR. Anti-interleukin 6 agents (tocilizumab, sarilumab) have shown consistent efficacy results, suggesting a particularly interesting steroid-sparing effect in the population under consideration. The safety profile appears acceptable. Other biologic targeted treatments are currently being evaluated. Anti-interleukin-6 agents may well have a place in the therapeutic strategy for PMR, particularly for patients with steroid-resistant disease or at high risk of complications of corticosteroid therapy.


Chronobiologic perspectives of black time--Accident risk is greatest at night: An opinion paper.
Simon Folkard in 1997 introduced the phrase black time to draw attention to the fact that the risk of driving accidents (DA) is greater during the night than day in usually diurnally active persons. The 24 h temporal pattern in DA entails circadian rhythms of fatigue and sleep propensity, cognitive and physical performance, and behavior that are controlled, at least in part, by endogenous clocks. This opinion paper extends the concept of black time to reports of excess nighttime accidents and injuries of workers and nocturnal occurrence of certain man-caused catastrophes. We explore the chronobiology of work-related black time accidents and injuries taking into account laboratory and field investigations describing, respectively, circadian rhythms in cognitive performance and errors and mistakes by employees in the conduct of routine occupational tasks. Additionally, we present results of studies pertaining to 24 h patterns of both the number and relative risk (number of events per h/number of workers exposed per h) of work-related accidents (WRA) and injuries (WRI) as well as indices of performance and alertness of a self-selected homogenous survivor cohort of French firefighters (FFs) to explore two possible explanations of black time, namely, 24 h variation in sleep propensity/drossiness characterized by a nocturnal peak and circadian rhythms in cognitive performance characterized by a nocturnal trough. We propose the 24 h pattern of WRA and WRI, particularly of FFs and other highly skilled self-selected cohorts, is more strongly linked to circadian rhythms of fatigue and sleepiness than cognitive performance. Other possible explanations--suppressed expression of circadian rhythms and/or unmasking of ultradian periodicities in cognitive performance in specific circumstances, e.g., highly stressful work, competitive, or life-threatening settings, are also discussed.


Structural and functional characterization of two unusual endonuclease III enzymes from Deinococcus radiodurans.
While most bacteria possess a single gene encoding the bifunctional DNA glycosylase Endonuclease III (EndoIII) in their genomes, Deinococcus radiodurans possesses three: DR2438 (DrEndoIII1), DR0289 (DrEndoIII2) and DR0982 (DrEndoIII3). Here we have determined the crystal structures of DrEndoIII1 and an N-terminally truncated form of DrEndoIII3 (DrEndoIII3Δ76). We have also generated a homology model of DrEndoIII2 and measured activity of the three enzymes. All three structures consist of two all α-helical domains, one of which exhibits a [4Fe-4S] cluster and the other a HhH-motif, separated by a DNA binding cleft, similar to previously determined structures of endonuclease III from Escherichia coli and Geobacillus stearothermophilus. However, both DrEndoIII1 and DrEndoIII3 possess an extended HhH motif with extra helical features and an altered electrostatic surface potential. In addition, the DNA binding cleft of DrEndoIII3 seems to be less accessible for DNA interactions, while in DrEndoIII1 it seems to be more open. Analysis of the enzyme activities shows that DrEndoIII2 is most similar to the previously studied enzymes, while DrEndoIII1 seems to be more distant with a weaker activity towards substrate DNA containing either thymine glycol or an abasic site. DrEndoIII3 is the most distantly related enzyme and displays no detectable activity towards these substrates even though the suggested catalytic residues are conserved. Based on a comparative structural analysis, we suggest that the altered surface potential, shape of the substrate-binding pockets and specific amino acid substitutions close to the active site and in the DNA interacting loops may underlie the unexpected differences in activity.


Abrasive Wear Resistance of Plasma-Nitrided Ti Enhanced by Ultrasonic Surface Rolling Processing Pre-Treatment.
The objective of the given work was to investigate abrasive wear behaviours of titanium (Ti) treated by ultrasonic surface rolling processing (USRP) pre-treatment and plasma nitriding (PN). Simulated lunar regolith particles (SLRPs) were employed as abrasive materials during characterization of tribological performances. The experimental results showed that SLRPs cause severe abrasive wear on Ti plasma-nitrided at 750 °C via the mechanism of micro-cutting. Due to the formation of a harder and thicker nitriding layer, the abrasive wear resistance of the Ti plasma-nitrided at 850 °C was enhanced, and its wear mechanism was mainly fatigue. USRP pre-treatment was effective at enhancing the abrasive wear resistance of plasma-nitrided Ti, due to the enhancement of the hardness and thickness of the nitride layer. Nevertheless, SLRPs significantly decreased the friction coefficient of Ti treated by USRP pre-treatment and PN, because the rolling of small granular abrasives impeded the adhesion of the worn surface. Furthermore, USRP pre-treatment also caused the formation of a dimpled surface with a large number of micropores which can hold wear debris during tribo-tests, and finally, polishing and rolling the wear debris resulted in a low friction coefficient (about 0.5).


[Fabry's disease associated with rheumatoid arthritis. Multisystemic crossroads].
Fabry's disease is a rare congenic disorder of glycosphingolipid catabolism resulting from deficient activity of the alpha galactosidasa. Is an X-linked disorder and in hemizygous males the activity of this enzyme is very low, resulting in severe manifestations. Fabry disease is confirmed by the lack alfa-galactosidase in serum. In the literature have been reported a few cases of coexistent Fabry's disease and connective disorders, but there is not cases of rheumatoid arthritis coexistent. This report describes a case of a female with Fabry's disease who vas subsequently diagnosed with rheumatoid arthritis. The suspect diagnosis was very important because the two disorders are multisystem and new symptoms could be attributed to Fabry's disease. The accumulation of lipids may results in numerous pathogenic autoantibodies, which could make immunocomplex. This is the potential pathogenic mechanisms explaining the association between Fabry's disease and autoimmune diseases.


Nematic Skyrmions in Odd-Parity Superconductors.
We study topological excitations in two-component nematic superconductors, with a particular focus on Cu_{x}Bi_{2}Se_{3} as a candidate material. We find that the lowest-energy topological excitations are coreless vortices: a bound state of two spatially separated half-quantum vortices. These objects are nematic Skyrmions, since they are characterized by an additional topological charge. The inter-Skyrmion forces are dipolar in this model, i.e., attractive for certain relative orientations of the Skyrmions, hence forming multi-Skyrmion bound states.


Annexin 2 is not required for human immunodeficiency virus type 1 particle production but plays a cell type-dependent role in regulating infectivity.
During assembly and budding of retroviruses, host cell proteins are incorporated into viral particles. Identification of virion-associated proteins may help pinpoint key cellular components required for virus production and function. The cellular protein annexin 2 (Anx2) is incorporated into HIV-1 particles, and knockdown of Anx2 has been reported to cause defects in Gag processing and infectivity of HIV-1 particles in macrophages. Here, we tested whether Anx2 was required for HIV-1 production in other cell types capable of producing HIV-1 virions. Endogenous Anx2 levels were knocked down by approximately 98% using lentivirus encoding short hairpin RNAs (shRNAs) or small interfering RNAs (siRNAs) targeting Anx2. Under these conditions, there was no reduction in HIV-1 virus-like particle (VLP) production in either COS-1, 293T, or Jurkat T cells or primary human monocyte-derived macrophages (MDMs). Murine embryonic fibroblasts derived from Anx2(-/-) mice produced the same levels of VLPs as matched cells from wild-type mice. The calcium-mediated spike in VLP production still occurred in Anx2-depleted COS-1 cells, and there was no apparent alteration in the intracellular Gag localization. Overexpression of Anx2 in trans had no effect on Gag processing or VLP production. Neither Anx2 depletion nor Anx2 overexpression altered the infectivity of HIV-1 particles produced by COS-1 or 293T cells. However, supernatants containing virus from Anx2 siRNA-treated primary human MDMs exhibited decreased infectivity. These data indicate that Anx2 is not required for HIV-1 assembly or Gag processing but rather plays a cell type-dependent role in regulating production of infectious HIV-1 by macrophages.


Principles of managing the high-risk pregnant patient.
The family physician has opportunities for reducing perinatal morbidity and mortality. The potential for a high-risk pregnancy must be recognized. Important elements in the management of a high-risk pregnancy include determination of fetal age, ultrasound, estriol determination, contraction stress test, nonstress test, lecithin/sphingomyelin ratio and scalp blood pH measurement. Electronic fetal monitoring, including direct techniques, should be used in the course of labor.


Cognitive Skills Training for Homeless Transition-Age Youth: Feasibility and Pilot Efficacy of a Community Based Randomized Controlled Trial.
Cognitive impairments are common in homeless youth and negatively impact academic and vocational outcomes. We examined the feasibility and efficacy of cognitive interventions provided to 18- to 22-year-old homeless youth living in urban supportive housing. Ninety-one homeless youth were randomized to receive either targeted cognitive training (cognitive remediation) or general cognitive activation (computer skills training). Cognitive and psychological outcomes were assessed at baseline, after 13 and 26 sessions, and 1 month postintervention. A high dropout rate highlighted the feasibility challenges of treating this population. Intent-to-treat analysis found significant improvements across groups in specific and global measures of cognition and psychological distress, with no significant group differences. Transition-age homeless youth show improvements in cognitive and psychological functioning when engaged in interventions that address their cognitive development. This speaks to the malleability of cognitive skills in this cohort and lays the groundwork for future research to address their cognitive health.


Regulation of intron function: efficient splicing in vivo of a bacterial group II intron requires a functional promoter within the intron.
Conjugative transfer of the Lactococcus lactis plasmid pRS01 requires splicing of a group II intron, Ll.ltrB, for accurate translation of the mRNA for the exon gene ltrB. The protein product of ltrB is a conjugative relaxase, essential for pRS01 transfer. Using a molecular technique for the identification of transcription initiation sites in bacteria, a functional promoter within Ll.ltrB was identified upstream from the gene for the intron-encoded protein (IEP) LtrA. LtrA is required for efficient splicing of Ll.ltrB in vivo. Mutation of the ltrA promoter dramatically reduced the steady-state level of ltrA mRNA, LtrA, intron splicing and conjugation in L. lactis. These effects could be relieved by expression in trans of the ltrA gene cloned under the control of an inducible promoter. These results suggest that the ltrA mRNAs are translated inefficiently. We hypothesize that this bacterial intron, in contrast to previously studied group II introns in eukaryotes, requires a promoter within the intron to regulate ltrA expression and to produce an adequate level of the protein in the cell for efficient splicing.


Pre-heart failure at 2D- and 3D-speckle tracking echocardiography: A comprehensive review.
Chronic heart failure (CHF) has different stages and includes pre-HF (PHF), a state of high risk of developing myocardial dysfunction and advanced CHF. Some major behavioral risk factors of PHF might predispose to biological risk factors such as obesity, diabetes mellitus, dyslipidemia, hypertension, myocardial infarction, and cardiomyopathy. These risk factors damage the myocytes leading to fibrosis, apoptosis, cardiac hypertrophy, along with alterations in cardiomyocyte' size and shape. A condition of physiological subcellular remodeling resulting into a pathological state might be developed, conducting to PHF. Both PHF and heart failure (HF) are associated with the activation of phospholipases and protease, mitochondrial dysfunction, oxidative stress and development of intra-cellular free Ca2+ [Ca2+ ]i overloading to an elevation in diastolic [Ca2+ ]i . Simultaneously, cardiac gene expression is activated leading to further molecular, structural and biochemical changes of the myocardium. The sub-cellular remodeling may be intimately involved in the transition of cardiac hypertrophy to heart failure. 2D- and 3D-speckle tracking echocardiography (STE) have been used to quantify regional alterations of longitudinal strain and area strain, through their polar projection, which permits a further assessment of both sites and degrees of myocardial damage. The examination of strain can identify sub-clinical cardiac dysfunction or cardiomyocyte remodeling. During remodeling of the myocardium cardiac strain is attenuated, therefore it is an indicator of disease assessment.


Indirect evidence and the poverty of the stimulus: the case of anaphoric one.
It is widely held that children's linguistic input underdetermines the correct grammar, and that language learning must therefore be guided by innate linguistic constraints. Here, we show that a Bayesian model can learn a standard poverty-of-stimulus example, anaphoric one, from realistic input by relying on indirect evidence, without a linguistic constraint assumed to be necessary. Our demonstration does, however, assume other linguistic knowledge; thus, we reduce the problem of learning anaphoric one to that of learning this other knowledge. We discuss whether this other knowledge may itself be acquired without linguistic constraints.


Palliative care for patients with chronic obstructive pulmonary disease: current perspectives.
Chronic obstructive pulmonary disease (COPD) is a chronic respiratory illness with a myriad of disabling symptoms and a decline in the functional parameters that affect the quality of life. The mortality and morbidity associated with severe COPD is high and the patients are mostly housebound and in need of continuous care and support. The uncertain nature of its prognosis makes the commencement of palliative care and discussion of end-of-life issues difficult even in the advanced stage of the disease. This is often compounded by inadequate communication and counseling with patients and their relatives. The areas that may improve the quality of care include the management of dyspnea, oxygen therapy, nutritional support, antianxiety, and antidepressant treatment, and advance care planning. Hence, it is necessary to pursue a holistic care approach for palliative care services along with disease-specific medical management in all such patients to improve the quality of life in end-stage COPD.


Polymerase δ deficiency causes syndromic immunodeficiency with replicative stress.
Polymerase δ is essential for eukaryotic genome duplication and synthesizes DNA at both the leading and lagging strands. The polymerase δ complex is a heterotetramer comprising the catalytic subunit POLD1 and the accessory subunits POLD2, POLD3, and POLD4. Beyond DNA replication, the polymerase δ complex has emerged as a central element in genome maintenance. The essentiality of polymerase δ has constrained the generation of polymerase δ-knockout cell lines or model organisms and, therefore, the understanding of the complexity of its activity and the function of its accessory subunits. To our knowledge, no germline biallelic mutations affecting this complex have been reported in humans. In patients from 2 independent pedigrees, we have identified what we believe to be a novel syndrome with reduced functionality of the polymerase δ complex caused by germline biallelic mutations in POLD1 or POLD2 as the underlying etiology of a previously unknown autosomal-recessive syndrome that combines replicative stress, neurodevelopmental abnormalities, and immunodeficiency. Patients' cells showed impaired cell-cycle progression and replication-associated DNA lesions that were reversible upon overexpression of polymerase δ. The mutations affected the stability and interactions within the polymerase δ complex or its intrinsic polymerase activity. We believe our discovery of human polymerase δ deficiency identifies the central role of this complex in the prevention of replication-related DNA lesions, with particular relevance to adaptive immunity.


In vivo pH in metabolic-defective Ras-transformed fibroblast tumors: key role of the monocarboxylate transporter, MCT4, for inducing an alkaline intracellular pH.
We present an investigation of tumor pH regulation, designed to support a new anticancer therapy concept that we had previously proposed. Our study uses a tumor model of ras-transformed hamster fibroblasts, CCL39, xenografted in the thighs of nude mice. We demonstrate, for the first time, that genetic modifications of specific mechanisms of proton production and/or proton transport result in distinct, reproducible changes in intracellular and extracellular tumor pH that can be detected and quantified noninvasively in vivo, simultaneously with determinations of tumor energetic status and necrosis in the same experiment. The CCL39 variants used were deficient in the sodium/proton exchanger, NHE-1, and/or in the monocarboxylate transporter, MCT4; further, variants were deficient in glycolysis or respiration. MCT4 expression markedly increased the gradient between intracellular and extracellular pH from 0.14 to 0.43 when compared to CCL39 wild-type tumors not expressing MCT4. The other genetic modifications studied produced smaller but significant increases in intracellular and decreases in extracellular pH. In general, increased pH gradients were paralleled by increased tumor growth performance and diminished necrotic regions, and 50% of the CCL39 variant expressing neither MCT4 nor NHE-1, but possessing full genetic capacity for glycolysis and oxidative phosphorylation, underwent regression before reaching a 1-cm diameter. Except for CCL39 wild-type tumors, no significant HIF-1α expression was detected. Our in vivo results support a multipronged approach to tumor treatment based on minimizing intracellular pH by targeting several proton production and proton transport processes, among which the very efficient MCT4 proton/lactate co-transport deserves particular attention.


Drought stress had a predominant effect over heat stress on three tomato cultivars subjected to combined stress.
Abiotic stresses due to environmental factors could adversely affect the growth and development of crops. Among the abiotic stresses, drought and heat stress are two critical threats to crop growth and sustainable agriculture worldwide. Considering global climate change, incidence of combined drought and heat stress is likely to increase. The aim of this study was to shed light on plant growth performance and leaf physiology of three tomatoes cultivars ('Arvento', 'LA1994' and 'LA2093') under control, drought, heat and combined stress. Shoot fresh and dry weight, leaf area and relative water content of all cultivars significantly decreased under drought and combined stress as compared to control. The net photosynthesis and starch content were significantly lower under drought and combined stress than control in the three cultivars. Stomata and pore length of the three cultivars significantly decreased under drought and combined stress as compared to control. The tomato 'Arvento' was more affected by heat stress than 'LA1994' and 'LA2093' due to significant decreases in shoot dry weight, chlorophyll a and carotenoid content, starch content and NPQ (non-photochemical quenching) only in 'Arvento' under heat treatment. By comparison, the two heat-tolerant tomatoes were more affected by drought stress compared to 'Arvento' as shown by small stomatal and pore area, decreased sucrose content, ΦPSII (quantum yield of photosystem II), ETR (electron transport rate) and qL (fraction of open PSII centers) in 'LA1994' and 'LA2093'. The three cultivars showed similar response when subjected to the combination of drought and heat stress as shown by most physiological parameters, even though only 'LA1994' and 'LA2093' showed decreased Fv/Fm (maximum potential quantum efficiency of photosystem II), ΦPSII, ETR and qL under combined stress. The cultivars differing in heat sensitivity did not show difference in the combined stress sensitivity, indicating that selection for tomatoes with combined stress tolerance might not be correlated with the single stress tolerance. In this study, drought stress had a predominant effect on tomato over heat stress, which explained why simultaneous application of heat and drought revealed similar physiological responses to the drought stress. These results will uncover the difference and linkage between the physiological response of tomatoes to drought, heat and combined stress and be important for the selection and breeding of tolerant tomato cultivars under single and combine stress.


[Ion-exchange chromatography on cholesterol oxidase from Actinomyces lavendulae on a Biocarb-type carboxyl cationite].
Reversible sorption on the Biocarb-D carboxyl cationite of cholesterol oxidase extracted from the mycelium of Actinomyces lavendulae was examined. Sorption at pH 5.0 and desorption at pH 5.5 maintained 80% of the enzyme activity and increased seven-fold the specific activity as calculated per protein. By gel chromatography on Sephadex G-150 it was shown that the process was accompanied by a significant decrease in the number of inactive proteins.


Cardiovascular risk factors, exercise capacity and health literacy in patients with chronic ischaemic heart disease and type 2 diabetes mellitus in Germany: Baseline characteristics of the Lifestyle Intervention in Chronic Ischaemic Heart Disease and Type 2 Diabetes study.
Lifestyle interventions are a cornerstone in the treatment of chronic ischaemic heart disease (CIHD) and type 2 diabetes mellitus (T2DM). This study aimed at identifying differences in clinical characteristics between categories of the common lifestyle intervention targets BMI, exercise capacity (peak V̇O2) and health literacy (HL). Cross-sectional baseline characteristics of patients enrolled in the LeIKD trial (Clinicaltrials.gov NCT03835923) are presented in total, grouped by BMI, %-predicted peak V̇O2 and HL (HLS-EU-Q16), and compared to other clinical trials with similar populations. Among 499 patients (68.3±7.7 years; 16.2% female; HbA1c, 6.9±0.9%), baseline characteristics were similar to other trials and revealed insufficient treatment of several risk factors (LDL-C 92±34 mg/dl; BMI, 30.1±4.8 kg/m2; 69.6% with peak V̇O2<90% predicted). Patients with lower peak V̇O2 showed significantly higher (p < 0.05) CIHD and T2DM disease severity (HbA1c, CIHD symptoms, coronary artery bypass graft). Obese patients had a significantly higher prevalence of hypertension and higher triglyceride levels, whereas in patients with low HL both quality of life components (physical, mental) were significantly reduced. In patients with CIHD and T2DM, peak V̇O2, BMI and HL are important indicators of disease severity, risk factor burden and quality of life, which reinforces the relevance of lifestyle interventions.


[Superior vena cava syndrome: a study based on 81 cases].
Characterize the etiology, clinical picture, radiographic findings, usefulness of several diagnostic studies in establishing an etiologic diagnosis, and outcome after treatment of superior vena cava syndrome (SVCS). We reviewed the charts of all patients diagnosed of SVCS in our hospital between the years 1983 and 1996. Eighty-one patients (70 male, 11 female) with an average age of 61 years were included in the study. In 77 (95%) patients a neoplastic cause was diagnosed, mainly lung cancer (76%). The most common clinical findings were: feeling of fullness in the head (81%), edema of the face and arms (78%), jugular venous distention (75%), prominent venous pattern over the chest wall (73%), dyspnea (59%), and cough (37%). The average length of time from the onset of symptoms to the date of diagnosis was 28 days. The chest x-ray was abnormal in 78 (96%) cases: mediastinal widening (51%) and right hilar mass (48%) were the most common findings. The diagnosis of SVCS was evident (from clinical picture and chest roentgenogram) in 76 (94%) patients. In SVCS due to malignancy, the most productive investigations in establishing a pathologic diagnosis were bronchoscopy (54%) and biopsy of a superficial lymph node (18%). Partial or complete clinical response to treatment was achieved in seventy percent of the cases. SVCS is an uncommon entity and his most frequent cause is bronchogenic carcinoma. The characteristic clinical and radiographic findings allow to do a reliable diagnosis of the syndrome. Initially, clinical response (partial or complete) to treatment is achieved in most patients.


Evaluating respondent-driven sampling as an implementation tool for universal coverage of antiretroviral studies among men who have sex with men living with HIV.
The TRUST model based on experimental and observational data posits that integration of HIV prevention and universal coverage of antiretroviral treatment at a trusted community venue provides a framework for achieving effective reduction in HIV-related morbidity and mortality among men who have sex with men (MSM) living with HIV, as well as reducing HIV incidence. The analyses presented here evaluate the utility of respondent-driven sampling as an implementation tool for engaging MSM in the TRUST intervention. The TRUST integrated prevention and treatment model was established at a trusted community center serving MSM in Abuja, Nigeria. Five seeds have resulted in 3-26 waves of accrual between March 2013 and August 2014, with results presented here characterizing HIV burden and engagement in HIV care for 722 men across study recruitment waves. For analytic purposes, the waves were collapsed into 5 groups: 4 equally spaced (0-4, 5-9, 10-14, and 15-19) and 1 ranging from the 20th to the 26th wave with significance assessed using Pearson χ2 test. In earlier waves, MSM were more likely to have reported testing for HIV (82.9% in waves 0-4, 47.7% in waves 20-26; P < 0.01). In addition, biologically confirmed HIV prevalence decreased from an average of 59.1% to 42.9% (P < 0.05) in later waves. In earlier waves, about 80% of participants correctly reported their HIV status as compared with less than 25% in the later waves (P < 0.01). Finally, participants reporting being on ART decreased from 50% to 22.2% in later waves (P < 0.01). Implementation science studies focused on demonstrating impact of universal HIV treatment programs among people living with HIV necessitate different accrual methods than those focused on preventing HIV acquisition. Here, respondent-driven sampling was shown to be an efficient method for reaching marginalized populations of MSM living with HIV in Nigeria, and engaging them in universal HIV treatment services.


Understanding how college students characterise and cope with chronic pain: a thematic analysis of expressive writing samples.
College students who experience chronic pain are a frequently overlooked population. This research attempts to provide insight into the language that college students use to describe their experiences with chronic pain, challenges they face and coping strategies they use. Over the course of 4 consecutive days, participants responded to an expressive writing prompt asking them to reflect on their emotions and thoughts related to being a college student with chronic pain. Writing samples were then analysed to identify themes pertaining to words with a positive or negative emotional valence, terms used to characterise pain, metaphors used to describe pain, challenges faced, and positive and negative coping strategies used. Results showed that participants were more likely to use negatively valenced words than positively valenced words to describe their pain. Several common words and phrases were used to characterise pain, including metaphors. Finally, participants reported a number of challenges associated with their lived-experience of chronic pain, as well as positive and negative strategies used to cope with those challenges. These findings help to put into perspective the language used to understand, and attempt to cope with, the challenges faced by college students experiencing chronic pain. Research must continue to investigate the needs of this population so that they can be properly supported physically, emotionally, socially and academically.


Antimicrobial resistance in Salmonella enterica subspecies enterica serovar Dublin from dairy source calves in the central San Joaquin Valley, California (1998-2002).
The present study describes antimicrobial resistance patterns of Salmonella enterica subspecies enterica serovar Dublin (S. Dublin) in clinical submissions from calves and temporal and farm-type trends in antimicrobial resistance patterns of the isolates. A total of 300 isolates of S. Dublin were obtained from fecal or internal organs of calves fewer than 120 days of age originating from 84 dairies and 18 calf ranches from July 1998 to December 2002. The isolates were susceptibility tested to a panel of 10 antimicrobials using the disk diffusion assay. Temporal and farm-type trends in individual antimicrobial inhibition zone sizes were assessed and antimicrobial resistance patterns were described using cluster analysis. Isolates obtained from calf ranches compared with dairies exhibited decreased susceptibility to florfenicol, gentamicin, neomycin, sulfisoxazole, sulfamethoxazole/trimethoprim, and tetracycline. During the years 1998-2002, decreasing susceptibility was seen for ceftiofur, enrofloxacin, and sulfamethoxazole/trimethoprim. There were 20 different antimicrobial resistance patterns in the isolate set, indicating that S. Dublin has the ability to transfer and pick up resistance genes with relative ease. The trends seen in antimicrobial resistance in S. Dublin may likely be linked to antimicrobial drug use in young calves.


Rabies surveillance in the United States during 2019.
To provide epidemiological information on animal and human cases of rabies occurring in the United States during 2019 and summaries of 2019 rabies surveillance for Canada and Mexico. All animals submitted for laboratory diagnosis of rabies in the United States during 2019. State and territorial public health departments and USDA Wildlife Services provided data on animals submitted for rabies testing in the United States during 2019. Data were analyzed temporally and geographically to assess trends in domestic and wildlife rabies cases. During 2019, 53 jurisdictions submitted 97,523 animal samples for rabies testing, of which 94,770 (97.2%) had a conclusive (positive or negative) test result. Of these, 4,690 tested positive for rabies, representing a 5.3% decrease from the 4,951 cases reported in 2018. Texas (n = 565 [12.0%]), New York (391 [8.3%]), Virginia (385 [8.2%]), North Carolina (315 [6.7%]), California (276 [5.9%]), and Maryland (269 [5.7%]) together accounted for almost half of all animal rabies cases reported in 2019. Of the total reported rabid animals, 4,305 (91.8%) were wildlife, with raccoons (n = 1,545 [32.9%]), bats (1,387 [29.6%]), skunks (915 [19.5%]), and foxes (361 [7.7%]) as the primary species confirmed with rabies. Rabid cats (n = 245 [5.2%]) and dogs (66 [1.4%]) accounted for > 80% of rabies cases involving domestic animals in 2019. No human rabies cases were reported in 2019. The overall number of animal rabies cases decreased from 2018 to 2019. Laboratory diagnosis of rabies in animals is critical to ensure that human rabies postexposure prophylaxis is administered judiciously.


Regulation of energy metabolism by inflammation: a feedback response in obesity and calorie restriction.
Caloric restriction (CR), in the absence of malnutrition, delays aging and prevents aging-related diseases through multiple mechanisms. A reduction in chronic inflammation is widely observed in experimental models of caloric restriction. The low inflammation status may contribute to the reduced incidence of osteoporosis, Alzheimer's disease, cardiovascular diseases and cancer in the aging subjects. The association of caloric restriction with low inflammation suggests a role of energy accumulation in the origin of the chronic inflammation. This point is enforced by recent advances in obesity research. Abundant literature on obesity suggests that chronic inflammation is a consequence of energy accumulation in the body. The emerging evidence strongly supports that the inflammatory response induces energy expenditure in a feedback manner to fight against energy surplus in obesity. If this feedback system is deficient (Inflammation Resistance), energy expenditure will be reduced and energy accumulation will lead to obesity. In this perspective, we propose that an increase in inflammation in obesity promotes energy expenditure with a goal to get rid of energy surplus. A decrease in inflammation under caloric restriction contributes to energy saving. Inflammation is a mechanism for energy balance in the body. Inflammation resistance will lead to obesity. We will review the recent literature in support of the viewpoints.


Cytostatics for acromegaly. Marked improvement in a patient with an invasive pituitary tumour.
A 21-year-old woman suffering from acromegaly was treated with transsphenoidal subtotal hypophysectomy (microscopy: acidophilic adenoma), followed by x-ray and bromocriptine therapy. Seven years later she was re-operated because of a partial bitemporal loss of vision, intracranial hypertension, and regrowth of the pituitary tumour seen on CT-scan. A large part of the invasive suprasellar tumour was then removed by transcranial approach. The neurosurgery was followed by cobalt radiotherapy and bromocriptine administration. Two years later, symptoms and signs of tumour growth reappeared. Administration of cytostatics, such as doxorubicin (Adriamycin) and lomustine (Belustine), resulted in distinct clinical improvement associated with a seven-fold decrease in the serum growth hormone concentration. The visual field became normal and the intracranial mass on a CT scan decreased markedly. As a result the patient was able to resume work.


[Hereditary cerebellar ataxias: from hammer to genetics].
The hereditary ataxias comprise a complex group of neurological disorders involving the cerebellum and its connections. Several classifications based on clinical and/or pathological data have been only partially successful. Recent progress in molecular genetics has identified the genic loci of hereditary ataxias and has allowed a more precise diagnosis of distinct genetic diseases. Trinucleotide repeat expansions has been recognized as a mechanism of disease in some autosomal dominant spinocerebellar ataxias (ADCA) (SCA1 to SCA7), including Machado-Joseph disease/SCA3, probably the most common form of ADCA in South Brazil, and Friedreich ataxia (GAA expansion-chromosome 9p). Familial alpha-tocopherol deficiency (chromosome 8q) may have a Friedreich ataxia phenotype and responds to the oral supplementation with vitamin E. Familial episodic ataxias with (EA1-chromosome 12p) and without (chromosome 19p-EA2) myokimia were identified, the first one caused by point mutations in the gene encoding the KCNA1 potassium voltage-gated channel. The gene responsible for ataxia-teleangiectasia (chromosome 11q) was found to encode a putative DNA binding protein kinase (ATM), related to the cell cycle control. One to 3% of the population are heterozygotic ATM gen carry and pose a higher risk of cancer when exposed to ionizing radiation. Molecular biology has provided us with useful tools to diagnosis and genetic counseling and, hopefully, will provide us with a better understanding of the pathogenesis and eventual treatment of the several forms of hereditary ataxias.


Electronic-cigarette smoke induces lung adenocarcinoma and bladder urothelial hyperplasia in mice.
Electronic-cigarettes (E-cigs) are marketed as a safe alternative to tobacco to deliver the stimulant nicotine, and their use is gaining in popularity, particularly among the younger population. We recently showed that mice exposed to short-term (12 wk) E-cig smoke (ECS) sustained extensive DNA damage in lungs, heart, and bladder mucosa and diminished DNA repair in lungs. Nicotine and its nitrosation product, nicotine-derived nitrosamine ketone, cause the same deleterious effects in human lung epithelial and bladder urothelial cells. These findings raise the possibility that ECS is a lung and bladder carcinogen in addition to nicotine. Given the fact that E-cig use has become popular in the past decade, epidemiological data on the relationship between ECS and human cancer may not be known for a decade to come. In this study, the carcinogenicity of ECS was tested in mice. We found that mice exposed to ECS for 54 wk developed lung adenocarcinomas (9 of 40 mice, 22.5%) and bladder urothelial hyperplasia (23 of 40 mice, 57.5%). These lesions were extremely rare in mice exposed to vehicle control or filtered air. Current observations that ECS induces lung adenocarcinomas and bladder urothelial hyperplasia, combined with our previous findings that ECS induces DNA damage in the lungs and bladder and inhibits DNA repair in lung tissues, implicate ECS as a lung and potential bladder carcinogen in mice. While it is well established that tobacco smoke poses a huge threat to human health, whether ECS poses any threat to humans is not yet known and warrants careful investigation.


Low frequency of p16(INK4a) alterations in insulinomas.
The molecular mechanisms contributing to the tumorigenesis of insulinomas are poorly understood. Disruption of the cell cycle due to inactivation of the p16(INK4a) tumor-suppressor gene was identified in a variety of human tumors, including gastrinomas and nonfunctioning endocrine pancreatic carcinomas. In this study the role of p16(INK4a) in the tumorigenesis of insulinomas was evaluated. Seventeen insulinomas (14 benign, 3 malignant) were analyzed for genetic alterations in the p16(INK4a) tumor-suppressor gene by SSCP, PCR-based deletion and methylation-specific assays. p16 expression was determined by immunohistochemistry. One malignant insulinoma showed a homozygous deletion of p16(INK4a) and another two benign insulinomas revealed aberrant methylation of the p16(INK4a) promoter region. All three tumors lacked p16 expression according to immunohistochemistry. None of the insulinomas carried intragenic p16(INK4a) mutations. In total, 17% of insulinomas had p16(INK4a) alterations. The p16(INK4a) tumor-suppressor gene contributes to tumorigenesis in only a small subset of insulinomas.


U"AI" Testing: User Interface and Usability Testing of a Chest X-ray AI Tool in a Simulated Real-World Workflow.
Purpose: To observe interactions of practicing radiologists with a chest x-ray AI tool and evaluate its usability and impact on workflow efficiency. Methods: Using a simulated clinical workflow and remote multi-monitor screensharing, we prospectively assessed the interactions of 10 staff radiologists (5-33 years of experience) with a PACS-embedded, regulatory-approved chest x-ray AI tool. Qualitatively, we collected feedback using a think-aloud method and post-testing semi-structured interview; transcript themes were categorized by: (1) AI tool features, (2) deployment considerations, and (3) broad human-AI interactions. Quantitatively, we used time-stamped video recordings to compare reporting and decision-making efficiency with and without AI assistance. Results: For AI tool features, radiologists appreciated the simple binary classification (normal vs abnormal) and found the heatmap essential to understand what the AI considered abnormal; users were uncertain of how to interpret confidence values. Regarding deployment considerations, radiologists thought the tool would be especially helpful for identifying subtle diagnoses; opinions were mixed on whether the tool impacted perceived efficiency, accuracy, and confidence. Considering general human-AI interactions, radiologists shared concerns about automation bias especially when relying on an automated triage function. Regarding decision-making and workflow efficiency, participants began dictating 5 seconds later (42% increase, P = .02) and took 14 seconds longer to complete cases (33% increase, P = .09) with AI assistance. Conclusions: Radiologist usability testing provided insights into effective AI tool features, deployment considerations, and human-AI interactions that can guide successful AI deployment. Early AI adoption may increase radiologists' decision-making and total reporting time but improves with experience.


A Research Translation, Implementation and Impact Strategy for the Australian Healthy Environments and Lives (HEAL) Research Network.
Healthy Environments And Lives (HEAL) is the Australian national research network established to support improvements to health, the Australian health system, and the environment in response to the unfolding climate crisis. The HEAL Network comprises researchers, community members and organisations, policymakers, practitioners, service providers, and other stakeholders from diverse backgrounds and sectors. HEAL seeks to protect and improve public health, reduce health inequities and inequalities, and strengthen health system sustainability and resilience in the face of environmental and climate change, all with a commitment to building on the strengths, knowledge, wisdom, and experience of Aboriginal and Torres Strait Islander people, culture, and communities. Supporting applied research that can inform policy and practice, and effective research translation, implementation, and impact are important goals across the HEAL Network and essential to achieve its intended outcomes. To aid translation approaches, a research translation, implementation, and impact strategy for the HEAL Network was developed. The strategy has been created to inform and guide research translation across HEAL, emphasising communication, trust, partnerships, and co-design with communities and community organisations as well as the decision-makers responsible for public policies and programs. Development of the strategy was guided by research translation theory and practice and the Health in All Policies and Environment in All Policies frameworks. As described in this paper, the strategy is underpinned by a set of principles and outlines preliminary actions which will be further expanded over the course of the HEAL Network's activities. Through these actions, the HEAL Network is well-positioned to ensure successful research translation and implementation across its program of work.


Determination of drought tolerance related traits in Helianthus argophyllus, Helianthus annuus, and their hybrids.
Drought is a major constraint for sunflower (Helianthus annuus) production worldwide. Drought tolerance traits have been identified in the related wild species Helianthus argophyllus. This study was initiated to develop sunflower drought-tolerant genotypes by crossing cultivated sunflower with this species and analyze drought tolerance traits in the H. annuus and H. argophyllus populations, H. annuus intraspecific hybrids, and H. annuus × H. argophyllus interspecific hybrids along with the commercial hybrid Hysun-33 under three stress regimes: exogenous application of ABA, both by foliar spray and irrigation, and 5% PEG-induced osmotic stress. H. argophyllus populations had a significantly lower leaf area and higher water-use efficiency and leaf cuticular wax content under all treatments, and maintained a higher net photosynthetic rate and stomatal conductance under osmotic stress. Small leaf area and high cuticular waxes content of the wild species were, however, not inherited in interspecific hybrids which suggested for selection in F2 for these traits. Therefore, transgressive plants were selected in the F2 population to establish F3 plant progenies with silver-leafed canopy of H. argophyllus which showed higher achene yield under stress condition. These results are discussed with a view to using H. argophyllus to improve drought tolerance in cultivated sunflower.


Role of β-conglycinin and glycinin subunits in the pH-shifting-induced structural and physicochemical changes of soy protein isolate.
Soy β-conglycinin (7S) and glycinin (11S) were incubated up to 4 h in acidic (pH 1.5 to 3.5) or alkaline (pH 10 to 12) solutions to induce protein structural unfolding followed by refolding 1 h at pH 7.0, a process known as pH-shifting. The pH-shifting markedly increased (P < 0.05) emulsifying activity of 11S and to a lesser extent 7S; the former also produced more uniform oil droplets. The emulsifying activity improvements were accompanied by a significant rise in protein surface hydrophobicity, slight loss of the secondary structure (circular dichroism), and substantial dissociation of disulfide-linked basic and acidic 11S subunits. The findings suggested that 11S globulins of soy protein isolate (SPI) were more responsive to pH-shifting treatments than were 7S globulins, and the resulting emulsifying activity enhancements of 11S, in parallel with that of SPI, were indicative of its determinant role in the overall emulsifying properties of pH-shifting-treated SPI. Extreme alkaline (pH 12) and acidic (pH 1.5) medium treatments can significantly modify the structure and enhance the emulsifying properties of both β-conglycinin and glycinin components of SPI. The functionality improvement by the pH processes is more remarkable for the glycinin protein fraction. Therefore, SPI enriched with glycinin seems to be particularly suitable for extreme acidic or alkaline processes to produce surface-active functional ingredients for food applications.


An Evaluation of Family-Centered Rounds in the PICU: Room for Improvement Suggested by Families and Providers.
To identify areas for improvement in family-centered rounds from both the family and provider perspectives. Prospective, cross-sectional mixed-methods study, including an objective measure (direct observation of family-centered rounds) and subjective measures (surveys of English-speaking families and providers) of family-centered rounds. PICU in a single, tertiary children's hospital. Families of children admitted to the PICU, physicians, and nurses. None. Two hundred thirty-two family-centered round encounters were observed over a 10-week period. Family-centered round encounters averaged 10.5 minutes per child. Multivariable regression analysis revealed that family presence was independently associated with length of family-centered rounds (p < 0.002) despite family talk time accounting for an average of 25 seconds (4%) of the encounter. Non-English-speaking families were less likely to attend family-centered rounds compared with English-speaking families even when physically present at the patient's bedside (p < 0.001). Most commonly families and providers agreed that family-centered rounds keep the family informed and reported positive statements about family presence on family-centered rounds; however, PICU fellows did not agree that families provided pertinent information and nurses reported that family presence limited patient discussions. The primary advice families offered providers to improve family-centered rounds was to be more considerate and courteous, including accommodating family schedules, minimizing distractions, and limiting computer viewing. Family presence increased the length of family-centered rounds despite a small percentage of time spoken by families, suggesting longer rounds are due to changes in provider behavior when families are present. Also, non-English-speaking families may need more support to be able to attend and benefit from family-centered rounds. Lastly, in an era of full family-centered rounds acceptance, families and most providers, except fellows, report benefit from family presence during family-centered rounds. However, providers should be aware of the perception of their behaviors to optimize the experience for families.


Long-term resistance to simulated climate change in an infertile grassland.
Climate shifts over this century are widely expected to alter the structure and functioning of temperate plant communities. However, long-term climate experiments in natural vegetation are rare and largely confined to systems with the capacity for rapid compositional change. In unproductive, grazed grassland at Buxton in northern England (U.K.), one of the longest running experimental manipulations of temperature and rainfall reveals vegetation highly resistant to climate shifts maintained over 13 yr. Here we document this resistance in the form of: (i) constancy in the relative abundance of growth forms and maintained dominance by long-lived, slow-growing grasses, sedges, and small forbs; (ii) immediate but minor shifts in the abundance of several species that have remained stable over the course of the experiment; (iii) no change in productivity in response to climate treatments with the exception of reduction from chronic summer drought; and (iv) only minor species losses in response to drought and winter heating. Overall, compositional changes induced by 13-yr exposure to climate regime change were less than short-term fluctuations in species abundances driven by interannual climate fluctuations. The lack of progressive compositional change, coupled with the long-term historical persistence of unproductive grasslands in northern England, suggests the community at Buxton possesses a stabilizing capacity that leads to long-term persistence of dominant species. Unproductive ecosystems provide a refuge for many threatened plants and animals and perform a diversity of ecosystem services. Our results support the view that changing land use and overexploitation rather than climate change per se constitute the primary threats to these fragile ecosystems.


Fowler's syndrome and pregnancy.
We present a favourable outcome of a pregnant woman with underlying Fowler's syndrome. This is a rare disorder affecting young women with painless urinary retention, and as there is no known cure, the main concern is to ensure adequate bladder emptying. Our patient went through self-catheterisation and suprapubic catheters after which she was finally successfully managed on a sacral nerve stimulator (SNS). Upon getting pregnant the stimulator was switched off due to safety concerns, and from then on, bladder emptying was managed with a suprapubic catheter (which was regularly changed over the gestation). The main concern thereafter was recurrent hospital admissions with urinary tract infections and pelvic pain requiring parenteral antibiotics and analgesia. She underwent an uncomplicated elective caesarean section at 36 weeks and delivered a healthy female infant. The SNS was successfully reactivated in the postnatal phase, and the suprapubic catheter was removed upon achieving adequate urethral output.


Surface functionalization by covalent immobilization of an innovative carvacrol derivative to avoid fungal biofilm formation.
Carvacrol, an aromatic terpenic compound, known to be antimicrobial was grafted onto gold surfaces via two strategies based on newly-synthesized cross-linkers involving either an ester bond which can be cleaved by microbial esterases, or a covalent ether link. Surface functionalizations were characterized at each step by reflection absorption infrared spectroscopy (RAIRS). The two functionalized gold samples both led to a loss of culturability of the yeast Candida albicans, higher than 65%, indicating that the activity of the freshly-designed surfaces was probably due to still covalently immobilized carvacrol. On the contrary, when a phenyl group replaced the terpenic moiety, the yeast culturability increased by about 30%, highlighting the specific activity of carvacrol grafted on the surfaces. Confocal microscopy analyses showed that the mode of action of the functionalized surfaces with the ester or the ether of carvacrol was, in both cases, fungicidal and not anti-adhesive. Finally, this study shows that covalently immobilization of terpenic compounds can be used to design promising antimicrobial surfaces.


Modulation of tumour necrosis factor production with desmuramyldipeptide analogues.
Some synthetic analogues of the immunomodulatory agent muramyl dipeptide (MDP), i.e. phthalimido- (LK-511, LK-413, LK-512, LK-423, LK-508), adamantyl- (LK-415, LK-517), 7-oxoalkyl-(LK-409) desmuramylpeptides were assessed for the tumour necrosis factor (TNF) inducing activity and the ability to modulate TNF production in in vitro phorbol 12-myristate 13-acetate (PMA) & ionomycin stimulated cultures of human peripheral blood mononuclear cells. A kinetic study over a 40-hour period indicated that desmuramyldipeptides were weak TNF inducers compared to romurtide, PMA & ionomycin or lipopolysaccharide. By contrast, they showed the potential to up- or down-regulate the production of TNF evoked by PMA & ionomycin, which was strongly dependent on the time of the stimulation. After 4h of stimulation, the TNF secretion was augmented by LK-508, LK-409 and LK-511, after 18 h by LK-409 and LK-423, and after 40 h by LK-423, LK-511, LK-415 and LK-512. However, LK-517 and LK-512 inhibited the secretion of TNF after the 18-h period.


The link between neurosteroids and syndromic/syndromal components of the mood spectrum disorders in women during the premenstrual phase.
Females with a lifetime diagnosis of major mood disorder (Bipolar Disorder BD, Major Depressive Disorder MMD) investigated during the luteal phase of their menstrual cycle and in a condition of clinical well-being showed higher blood serum concentrations of progesterone and allopregnanolone compared to healthy controls. Women with BD presented even higher levels than those affected by MDD. This study attempted to verify, in line with a dimensional approach, if the possible differences in neurohormonal levels may be directly linked to some syndromal clusters (dimensions) of the mood spectrum disorders indipendently of diagnosis. Premenstrual concentrations of allopregnanolone, THDOC, progesterone, and cortisol were measured in 3 groups of women: 17 BD and 14 MDD outpatients, and 16 control subjects. Psychiatric evaluation was performed with the SCID-I interview and the SCI-MOODS-SR questionnaire. The correlation between steroid levels and mood disorder syndromal cluster (SCI-MOODS-SR domains and sub-domains) was evaluated by means of analysis of main components with Varimax rotation and Kaiser's normalization (which provided for inclusion of all components with an Eigen value >1). Analysis of the main components evidenced the presence of 3 components: 1) mania, 2) depression both with mixed component 3) steroid + manic cognitivity and suicidal ideas. Levels of allopregnanolone and progesterone do not correlate with the association of the depressive and manic syndromes, but rather with mixed symptomatological aspects, and in particular with cognitive manic and depressive (with suicidal thoughts) dimensions. Further studies should be carried out to confirm these findings.


Analyzing the Curriculum of the Faculty of Medicine, University of Gezira using Harden's 10 questions framework.
Despite the importance of curriculum analysis for internal refinement of a programme, the approach for such a step in under-described in the literature. This article describes the analysis of the medical curriculum at the Faculty of Medicine, University of Gezira (FMUG). This analysis is crucial in the era of innovative medical education since introducing new curricula and curricular changes has become a common occurrence in medical education worldwide. The curriculum analysis was qualitatively approached using descriptive analysis and adopting Harden's 10 Questions of curriculum development framework approach. Answering Harden's questions reflects the fundamental curricular components and how the different aspects of a curriculum framework fit together. The key features highlighted in the curriculum-related material and literature have been presented. The analysis of the curriculum of FMUG reveals a curriculum with interactive components. Clear structured objectives and goals reflect the faculty's vision. The approach for needs assessment is based on a scientific ground, and the curriculum integrated contents have been set to meet national and international requirements. Adopting SPICES strategies helps FMUG and students achieve the objectives of the curriculum. Multiple motivated instructional methods are adopted, fostering coping with the programme objectives and outcomes. A wide range of assessment methods has been adopted to assess the learning outcomes of the curriculum correctly, reliably, and in alignment with the intended outcomes. The prevailing conducive educational environment of FMUG is favourable for its operation and profoundly influences the outcome of the programme. And there is a well-defined policy for curriculum management, monitoring and evaluation. Harden's 10 questions are satisfactorily addressed by the multi-disciplinary and well-developed FMUG curriculum. The current curriculum supports the well-written faculty missions and educational objectives. It presents a structured, conceptual framework that supports the validity of the assumption behind the curriculum. The curriculum enhances intellectual and academic pursuits and supports social accountability.


Combined Effect of Health Status and Primary Care Use on Participation in Cancer Screening: The CONSTANCES Cohort.
The combined association between primary care utilization and health status with breast cancer screening (BCS) and cervical cancer screening (CCS) remains unclear. Our aim was to identify women's profiles based on their health status and primary care utilization and study their associated adherence to BCS and CCS recommendations. Using data from the cohort of people visiting health screening centers (CONSTANCES) in France (2012-2015), we first identified women's profiles based on their health status (self-perceived health, physical, and mental health) and primary care utilization (visit to the General Practitioner [GP], uptake of blood tests) using a multiple correspondence analysis and a hierarchical cluster analysis. We then investigated the association of these profiles to BCS and CCS using logistic regression models adjusted for age, smoking status, sociodemographic and socioeconomic characteristics, and the regularity of gynecologist consultation. We identified five distinct profiles of women with contrasted participation in BCS (n = 14,122) and CCS (n = 27,120). In multivariate analyses, cancer screening participation increased from women with very good health and poor primary care utilization, to those with poor health and frequent visits to the GP, and those with very good health and average primary care utilization. The most favorable profiles regarding cancer screening rates were women with average-to-poor health and regular visits to the GP and uptake of blood tests. Our results suggest that policies aiming at increasing cancer screening participation should simultaneously account for women's use of primary care and health and consider more specific subgroups than what is usually done. Further research should investigate factors motivating cancer screening practice, such as women's beliefs regarding cancer screening and women's psychological characteristics.


Machine Learning Algorithms for Predicting and Risk Profiling of Cardiac Surgery-Associated Acute Kidney Injury.
Using a large national database of cardiac surgical procedures, we applied machine learning (ML) to risk stratification and profiling for cardiac surgery-associated acute kidney injury. We compared performance of ML to established scoring tools. Four ML algorithms were used, including logistic regression (LR), gradient boosted machine (GBM), K-nearest neighbor, and neural networks (NN). These were compared to the Cleveland Clinic score, and a risk score developed on the same database. Five-fold cross-validation repeated 20 times was used to measure the area under the receiver operating characteristic curve (AUC), sensitivity, and specificity. Risk profiles from GBM and NN were generated using Shapley additive values. A total of 97,964 surgery events in 96,653 patients were included. For predicting postoperative renal replacement therapy using pre- and intraoperative data, LR, GBM, and NN achieved an AUC (standard deviation) of 0.84 (0.01), 0.85 (0.01), 0.84 (0.01) respectively outperforming the highest performing scoring tool with 0.81 (0.004). For predicting cardiac surgery-associated acute kidney injury, LR, GBM, and NN each achieved 0.77 (0.01), 0.78 (0.01), 0.77 (0.01) respectively outperforming the scoring tool with 0.75 (0.004). Compared to scores and LR, shapely additive values analysis of black box model predictions was able to generate patient-level explanations for each prediction. ML algorithms provide state-of-the-art approaches to risk stratification. Explanatory modeling can exploit complex decision boundaries to aid the clinician in understanding the risks specific to individual patients.


Carbonic anhydrase 9 (CA9) expression in non-small-cell lung cancer: correlation with regulatory FOXP3+T-cell tumour stroma infiltration.
Low pH suppresses the proliferation and cytotoxic activity of CD8+ cytotoxic and natural killer lymphocytes. The hypoxia-regulated transmembrane protein, carbonic anhydrase CA9, converts carbon dioxide produced by the Krebs cycle to bicarbonate and protons that acidify the extracellular milieu. We examined whether CA9 is also involved in intratumoural immunosuppression pathways. A series of 98 tissue samples of primary non-small-cell lung carcinomas (NSCLC) from patients treated with surgery were analysed for the expression of CA9 and programmed-death ligand PD-L1 by cancer cells, and of FOXP3 by tumour-infiltrating lymphocytes (TILs). There was no direct association of CA9 with PD-L1 expression or the density of TILs in the tumour stroma, but CA9 was directly related to the extent of FOXP3+ TIL density (p = 0.008). Double-stratification survival analysis showed that patients with high CA9 expression and low TIL score had significantly poorer survival compared with all other groups (p < 0.04). In a multivariate analysis stage (p < 0.0001, HR 1.95, 95% CI: 1.3-2.7), TIL score (p = 0.05, HR 0.55, 95% CI: 0.2-1.0) was an independent prognostic variable of death events. CA9 expression by cancer cells is associated significantly with FOXP3+ regulatory T-cell abundance in the tumour stroma of NSCLC. The study provides a basis for testing CA9 as a marker of resistance to immune-checkpoint inhibitors and as a therapeutic target to enhance the efficacy of immunotherapy.


Changes in colorectal cancer incidence rates in young and older adults in the United States: what does it tell us about screening.
Colorectal cancer (CRC) incidence rates have increased among young adults and have decreased among older adults. We re-evaluated these trends using more recent data covering about 96 % of the United States population. Colorectal cancer incidence rates were abstracted from the National Program of Cancer Registries and the Surveillance Epidemiology and End Results analytic files for diagnosis years 1998-2009. We report rates for young adults (age <50 years) and for older adults (age 50 years or older) by four race/ethnicity groupings. We examined CRC incidence rates by stage at diagnosis, tumor subsite, and state. We calculated the correlation between state-specific CRC incidence and prevalence of colonoscopy reported in the Behavioral Risk Factor Surveillance System. Rectal cancer incidence rates increased from 1998 through 2009 among young non-Hispanic white adults and young blacks. Among older adults, CRC incidence rates decreased among all four race/ethnicity groupings and in all states. The decline was apparent for all stages and for all subsites. States with greater decreases in CRC incidence rates had higher colonoscopy screening rates. Rectal cancer is increasing among younger adults, for reasons largely unknown. Among older adults, CRC incidence continues to decrease, probably because of increasing uptake of colonoscopy screening. Decreases in CRC incidence are correlated with increased use of colonoscopy, indicating that CRC may be largely preventable through colonoscopy screening. Efforts to increase screening rates in underserved populations would help reduce health disparities associated with this type of cancer.


alpha-MSH production, receptors, and influence on neopterin in a human monocyte/macrophage cell line.
alpha-Melanocyte-stimulating hormone (alpha-MSH), a tridecapeptide derived from pro-opiomelanocortin, has potent antiinflammatory activity in laboratory animals. alpha-MSH inhibits nitric oxide production by murine macrophages, an influence believed to reflect activation of an autocrine circuit in these cells, one that is based on production and release of alpha-MSH and subsequent stimulation of melanocortin receptors. We found that THP-1 cells, human monocytic cells, produced alpha-MSH; this production was increased by interleukin-6, tumor necrosis factor a, or concanavalin A. These cells also expressed the gene for the human alpha-MSH receptor MC1. Unlike murine macrophages, THP-1 cells produced little nitrite in response to interferon-gamma (IFN-gamma) and lipopolysaccharide, and a-MSH inhibited this production only slightly. However, production of neopterin, a presumed primate homologue of nitric oxide in lower animals, was increased in THP-1 cells stimulated with INF-gamma plus TNF-alpha and alpha-MSH significantly inhibited this production. The evidence indicates that an autocrine regulatory circuit based on alpha-MSH occurs in human monocyte/macrophages much as in murine macrophages. alpha-MSH-induced modulation of specific inflammatory mediators/cytotoxic agents appears to differ depending on the importance of the mediators in the myelomonocytic cells of different species.


Effects of gestational exposure to ethane dimethanesulfonate in CD-1 mice: microtia and preliminary hearing tests.
Microtia is a reduction in pinna size, usually seen in humans in conjunction with other medical conditions. We report microtia in CD-1 mice after gestational exposure to ethane dimethanesulfonate (EDS), an alkylating agent and adult rat Leydig cell toxicant. Time-pregnant CD-1 mice were administered 0, 80, or 160 mg EDS/kg on gestation days (GD) 11-17, or 0 or 160 mg EDS/kg on GD 11-13, GD 13-15 or GD 15-17. Pinnae were measured on postnatal days (PND) 4, 8, 18, and 28; and were observed for detachment from birth through PND 8. Branchial-arch derived skeletal structures and histology of the pinna was examined on PND 4 and 24. Brainstem auditory evoked response (BAER) tests were carried out at approximately PND 160 to determine possible effects on hearing. All offspring of EDS-treated dams exhibited bilateral, dose-related decreases in pinna size. Gestational exposure during GD 11-13 produced smaller ears than during GD 13-15 or 15-17, but not as small as the GD 11-17 regimen. Ossification of other pharyngeal arch derivatives was delayed whereas histology was unremarkable. BAER analysis showed a decrease in the proportion of adult offspring producing a quantifiable response to varied auditory stimuli among EDS-treated litters. Gestational exposure to EDS affects pinna development in the mouse, with a broad period of sensitivity during the second half of gestation. Microtia induced by EDS may be associated with hearing deficits, suggesting functional importance of pinna size or additional effects of EDS on ear development not detected by morphological examination.


Structural Identification of Individual Helical Amyloid Filaments by Integration of Cryo-Electron Microscopy-Derived Maps in Comparative Morphometric Atomic Force Microscopy Image Analysis.
The presence of amyloid fibrils is a hallmark of more than 50 human disorders, including neurodegenerative diseases and systemic amyloidoses. A key unresolved challenge in understanding the involvement of amyloid in disease is to explain the relationship between individual structural polymorphs of amyloid fibrils, in potentially mixed populations, and the specific pathologies with which they are associated. Although cryo-electron microscopy (cryo-EM) and solid-state nuclear magnetic resonance (ssNMR) spectroscopy methods have been successfully employed in recent years to determine the structures of amyloid fibrils with high resolution detail, they rely on ensemble averaging of fibril structures in the entire sample or significant subpopulations. Here, we report a method for structural identification of individual fibril structures imaged by atomic force microscopy (AFM) by integration of high-resolution maps of amyloid fibrils determined by cryo-EM in comparative AFM image analysis. This approach was demonstrated using the hitherto structurally unresolved amyloid fibrils formed in vitro from a fragment of tau (297-391), termed 'dGAE'. Our approach established unequivocally that dGAE amyloid fibrils bear no structural relationship to heparin-induced tau fibrils formed in vitro. Furthermore, our comparative analysis resulted in the prediction that dGAE fibrils are structurally closely related to the paired helical filaments (PHFs) isolated from Alzheimer's disease (AD) brain tissue characterised by cryo-EM. These results show the utility of individual particle structural analysis using AFM, provide a workflow of how cryo-EM data can be incorporated into AFM image analysis and facilitate an integrated structural analysis of amyloid polymorphism.


Late rejection occurred in recipients who experienced acute cellular rejection within the first year after heart transplantation.
Serial endomyocardial biopsies (EMBs) are scheduled even several years after heart transplantation (HTx) to monitor for late rejection (LR). However, repeated EMBs are associated with an increased risk for fatal complications and decrease the quality of life of the recipient. We retrospectively analyzed clinical data from 42 adult recipients who had received HTx and were followed > 1 year at the University of Tokyo Hospital. Five recipients experienced LR at 1130 ± 157 days after HTx, and all 5 had experienced acute cellular rejection (ACR) with ISHLT grade ≥ 2R within the first year, which was treated with methylprednisolone pulse therapy (sensitivity, 1.000; specificity, 0.7027). Logistic regression analyses demonstrated that positive panel reactive antibody (PRA) was the only significant predictor for LR among all parameters at 1 year after HTx (P = 0.020, odds ratio 24.00). Among the 5 recipients with LR, LR occurred earlier in the two PRA positive recipients than in those with a negative PRA (981 ± 12 versus 1230 ± 110 days, P = 0.042). Among the perioperative parameters, gender mismatch [n = 13 (31%)] was the only significant predictor for ACR within the first year in logistic regression analyses (P = 0.042, odds ratio 4.200). In conclusion, the current schedule of serial EMBs should perhaps be reconsidered for recipients without any history of ACR within the first year due to their lower risk of LR.


WRN polymorphisms affect expression levels of plasminogen activator inhibitor type 1 in cultured fibroblasts.
Recessive mutations in WRN gene eliminate WRN protein function (helicase) and cause Werner syndrome. One of the most important clinical features of Werner syndrome patients are the premature onset and accelerated atherosclerosis process. Studies carried out on polymorphic WRN locus have shown that the alleles 1367R and 1074L confer protection for cardiovascular disease. Given that the levels of plasminogen activator inhibitor type 1 (PAI-1) were found to be significantly increased in Werner syndrome patients, is quiet possible that PAI-1 expression could be under regulation of WRN helicase. Therefore the purpose of this work was to evaluate the role of WRN polymorphism in modulating the expression of PAI-1. In order to accomplish our aim, an array of primary cultured fibroblasts from normal adult donors was genotyped for polymorphisms of both the WRN and PAI-1 loci. In addition, steady state levels of WRN and PAI-1 were measured by semi-quantitative RT-PCR assays in such cultures. To search for the potential relationship between the lack of WRN protein and PAI-1 expression, heterozygous cultures of fibroblasts (1367RC/1074LF; WRN genotype) were treated with a molecule of interference RNA against WRN messenger RNA (mRNA). We found that, carriers of 1367R and 1074L alleles of WRN shown to have low amounts of PAI-1 in plasma (7.56 +/- 5.02), as compared with carriers of 1367C and 1074F alleles (16.09 +/- 6.03). Moreover, fibroblasts from carriers with these alleles had low expression levels of PAI-1 mRNA. The treatment of heterozygous primary fibroblast cultures (1367RC/1074LF; WRN genotype) with iRNA against WRN mRNA caused PAI-1 overexpression. Treatment with normal PAI-1 inducers (TGFbeta, TNFalpha, or insulin) in these cultures and from those with genotypes 1367CC/1074FF and 1367RR/1074FL resulted in a genotype-dependent PAI-1 expression level. Our results suggest that polymorphisms in the WRN gene might have a significant role regulating PAI-1 levels in healthy individuals and "normal states" as well as acute or chronic stress, obesity, aging, acute inflammation, among others, where characteristic high levels of insulin, TNF alpha and TGFbeta, could favor PAI-1 high levels in carriers with polymorphic variants (C and F alleles), beyond the levels reached by carriers with other alleles (R and L alleles).


Pedestrian injuries and vehicle type in Maryland, 1995-1999.
Pedestrian deaths constitute the second largest category of motor vehicle deaths in the US. The present study examined how pedestrian injury is associated with vehicle type, while controlling for vehicle weight and speed.Police, trauma registry, and autopsy data were linked for injured pedestrians. Logistic regression analyses were performed to control for vehicle weight and speed. Outcomes included pedestrian mortality, injury severity score, and injuries to specific body regions. Compared to conventional cars, pedestrians hit by sport utility vehicles and pick-up trucks were more likely to have higher injury severity scores (odds ratio=1.48; 95% confidence interval: 1.18-1.87) and to die (odds ratio=1.72; 95% confidence interval: 1.31-2.28). These relationships diminished when vehicle weight and speed were controlled for. At lower speeds, pedestrians struck by sport utility vehicles, pick-up trucks, and vans were approximately two times as likely to have traumatic brain, thoracic, and abdominal injuries; at higher speeds, there was no such association. The overall increased danger sport utility vehicles and pick-up trucks present to pedestrians may be explained by larger vehicle masses and faster speeds. At slower speeds being hit by sport utility vehicles, and pick-up trucks, and vans resulted in specific injuries, indicating that vehicle design may contribute to different injury patterns.