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International audience; Ice losses from the Greenland and Antarctic ice sheets have accelerated since the 1990s, accounting for a significant increase in the global mean sea level. Here, we present a new 29-year record of ice sheet mass balance from 1992 to 2020 from the Ice Sheet Mass Balance Inter-comparison Exercise (IMBIE). We compare and combine 50 independent estimates of ice sheet mass balance derived from satellite observations of temporal changes in ice sheet flow, in ice sheet volume, and in Earth's gravity field. Between 1992 and 2020, the ice sheets contributed 21.0±1.9 mm to global mean sea level, with the rate of mass loss rising from 105 Gt yr−1 between 1992 and 1996 to 372 Gt yr−1 between 2016 and 2020. In Greenland, the rate of mass loss is 169±9 Gt yr−1 between 1992 and 2020, but there are large inter-annual variations in mass balance, with mass loss ranging from 86 Gt yr−1 in 2017 to 444 Gt yr−1 in 2019 due to large variability in surface mass balance. In Antarctica, ice losses continue to be dominated by mass loss from West Antarctica (82±9 Gt yr−1) and, to a lesser extent, from the Antarctic Peninsula (13±5 Gt yr−1). East Antarctica remains close to a state of balance, with a small gain of 3±15 Gt yr−1, but is the most uncertain component of Antarctica's mass balance. The dataset is publicly available at https://doi.org/10.5285/77B64C55-7166-4A06-9DEF-2E400398E452 (IMBIE Team, 2021).

Ice losses from the Greenland and Antarctic Ice Sheets have accelerated since the 1990s, accounting for a significant increase in global mean sea level. Here, we present a new 29-year record of ice sheet mass balance from 1992 to 2020 from the Ice Sheet Mass Balance Inter-comparison Exercise (IMBIE). We compare and combine 50 independent estimates of ice sheet mass balance derived from satellite observations of temporal changes in ice sheet flow, in ice sheet volume and in Earth’s gravity field. Between 1992 and 2020, the ice sheets contributed 21.0 ± 1.9 mm to global mean sea-level, with the rate of mass loss rising from 105 Gt yr-1 between 1992 and 1996 to 372 Gt yr-1 between 2016 and 2020. In Greenland, the rate of mass loss is 169 ± 9 Gt yr-1 between 1992 and 2020 but there are large inter-annual variations in mass balance with mass loss ranging from 86 Gt yr-1 in 2017 to 444 Gt yr-1 in 2019 due to large variability in surface mass balance. In Antarctica, ice losses continue to be dominated by mass loss from West Antarctica (-82 ± 9 Gt yr-1) and to a lesser extent from the Antarctic Peninsula (-13 ± 5 Gt yr-1). East Antarctica remains close to a state of balance (3 ± 15 Gt yr-1), but is the most uncertain component of Antarctica’s mass balance.

In this study, we examine the effect of transient mantle creep on the prediction of glacial isostatic adjustment (GIA) signals. Specifically, we compare predictions of relative sea level change from GIA from a set of Earth models in which transient creep parameters are varied in a simple Burgers model to a reference case with a Maxwell viscoelastic rheology. The model predictions are evaluated in two ways: first, relative to each other to quantify the effect of parameter variation, and second, for their ability to reproduce well-constrained sea level records from selected locations. Both the resolution and geographic location of the relative sea level observations determine whether the data can distinguish between model cases. Model predictions are most sensitive to the inclusion of transient mantle deformation in regions that are near-field and peripheral relative to former ice sheets. This sensitivity appears particularly true along the North American west coast in the region of the former Cordilleran Ice Sheet, which experienced rapid sea-level fall following deglaciation between 14-12 kyr BP. Relative to the Maxwell case, Burgers models better reproduce this rapid phase of regional postglacial sea level fall. As well, computed goodness-of-fit values in this region show a clear preference for models where transient deformation is present in the whole or lower mantle, and for models where the rigidity of the Kelvin element is weakened relative to the rigidity of the Maxwell element. In contrast, model predictions of relative sea-level change in the far-field show little or weak sensitivity to the inclusion of transient deformation.

While cyclin dependent kinases (CDKs) were originally implicated in cell cycle regulation, several CDKs including CDK7, CDK8, and CDK9 play critical roles in transcription through regulation of RNA polymerase II (Pol II) activity. In addition to expected phosphosite targets associated with DNA-binding transcription factors, chromatin regulators, or other known regulators of pol II activity, a phospho-proteomics study on a CDK8/19 inhibitor (Cortistatin A) also identified DNA replication and repair proteins, i.e., namely BRCA1 and MDC1, as substrates for CDK8. RVU120, a first in-class CDK8/19 inhibitor in phase I clinical trial demonstrated preclinical efficacy in acute myeloid leukemia PDX models. A robust relationship has also been observed between the exposure to RVU120 and inhibition of pSTAT5, a pharmacodynamic marker, in solid tumor indications in a recent clinical trial. Additionally, preclinical data suggested RVU120 reduced expression of genes related to interferon-related DNA damage signature (IRDS) suggesting a potential role of CDK8 in DNA repair signaling. CDK8 loss was identified as a synthetic lethal hit in two internal, orthogonal screens that surveyed targets which can synergize with niraparib inhibition. Niraparib inhibits PARP enzymatic activity and promotes PARP trapping on ssDNA breaks which lead to replication stress-induced dsDNA breaks that require BRCA-dependent homologous recombination repair (HRR). Combination with other therapeutic agents is anticipated to improve the efficacy of niraparib in HR proficient (HRP) and BRCA1/2.WT tumors. This work describes our proof-of-concept data from in vitro validation of combination synergy response of RVU120 with niraparib in a panel of 129 HRR.WT and HRR mutant cancer cell lines across ovarian, breast, lung, and colorectal cancer indications. Our results indicated that the niraparib+RVU120 combination was synergistic, and a strong synergy response, i.e., a SS (Synergy Score) ≥ 5 was detected by Loewe modelling across the entire surface of the combination matrix in ~26% of Horizon Discovery OncoSignatureTM panel cell lines that were largely resistant to niraparib monotherapy. The study findings indicated the potential of CDK8/19 inhibitors to enhance niraparib activity in both HRR.WT and HRR mutant populations. The potential benefit of CDK8/19i+niraparib combination will be further explored in PARPi-resistant models. Molecular studies are underway to explore potential biomarkers associated with synergy response, and to analyze the downstream effects on DNA Damage Response. Citation Format: Karen A. Simon-Crevatis, Shailesh R. Metkar, Justin T. Hardcastle, Keith Jansson, Usman Shabon, Rajarshi Choudhury, Euan A. Stronach, Kevin G. Coleman, Asli Muvaffak. CDK8 inhibition potentiates the efficacy of niraparib in homologous recombination proficient cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6175.

Com o crescimento, a função respiratória na criança revela modificações ní­tidas da função pulmonar. Com o objetivo de avaliar a capacidade vital e relaciona-la com o peso, estatura e idade em crianças do sexo masculino na faixa etária de 7 a 11 anos que freqüentavam uma escola particular em Itajaí­, SC, foi coletado o valor da capacidade vital (CV), através da ventilometria, o peso e a estatura. O valor médio da capacidade vital foi 2,38 litros ± 0,11. O coeficiente de correlação simples da estatura em relação a CV foi de r = 0,694; idade em relação a CV, r = 0,644; peso em relação a CV, r = 0,642. O coeficiente de correlação simples evidenciou que os volumes pulmonares têm relação com as variáveis estatura, idade e peso. A estatura é a variável que mais interfere em volumes pulmonares e apesar de escassa, a literatura disponí­vel confirmou um volume normal para esta faixa etária.Palavras-chave: capacidade vital, ventilometria, crescimento.

Purpose Hermansky-Pudlak syndrome (HPS) is a rare inherited disorder with ten reported genetic types; each type has defects in subunits of either Adaptor Protein-3 complex or Biogenesis of Lysosome-related Organelles Complex (BLOC)-1, -2, or -3. Very few patients with BLOC-1 deficiency (HPS-7, -8, and -9 types) have been diagnosed. We report results of comprehensive clinical testing and molecular analyses of primary fibroblasts from a new case of HPS-7. Results A 6-year old Paraguayan male presented with hypopigmentation, ocular albinism, nystagmus, reduced visual acuity, and easy bruising. He also experienced delayed motor and language development as a very young child; head and chest trauma resulted in intracranial hemorrhage with subsequent right hemiparesis and lung scarring. There was no clinical evidence of immunodeficiency or colitis. Whole mount transmission electron microscopy revealed absent platelet delta granules; platelet aggregation testing was abnormal. Exome sequencing revealed a homozygous nonsense mutation in the Dystrobrevin binding protein 1 (DTNBP1) gene [ NM_032122.4 : c.307C > T; p.Gln103*], previously reported in a Portuguese adult. The gene encodes the dysbindin subunit of BLOC-1. Dysbindin protein expression was negligible in our patient's dermal fibroblasts, while his DTNBP1 mRNA level was similar to that of a normal control. Conclusions Comprehensive clinical evaluation of the first pediatric case reported with HPS-7 reveals oculocutaneous albinism and platelet storage pool deficiency; his phenotype is consistent with findings in other patients with BLOC-1 disorders. This patient's markedly reduced Dysbindin protein expression in HPS-7 resulted from a mechanism other than nonsense mediated decay.

Combined alpha-delta platelet storage pool deficiency is characterized by the absence or reduction in the number of both alpha granules and dense bodies. This disorder can have variable severity as well as a variable inheritance pattern. We describe two patients from unrelated families with combined alpha-delta storage pool deficiency due to mutations in GFI1B, a zinc finger protein known to act as a transcriptional repressor of various genes. We demonstrate that this disease is associated with either a heterozygous mutation (de novo or familial) abrogating the binding of the zinc fingers with the promoter of its target genes, or by hypomorphic biallelic mutations in GFI1B leading to autosomal recessive inheritance.

Purpose Hemophilia B, an X-linked disease, manifests with recurrent soft tissue bleeding episodes. Hermansky-Pudlak syndrome, a rare autosomal recessive disorder, is characterized by oculocutaneous albinism and an increased tendency to bleed due to a platelet storage pool defect. We report a novel mutation in HPS6 in a Caucasian man with hemophilia B and oculocutaneous albinism. Results The patient was diagnosed with hemophilia B at age 4months due to recurrent soft tissue bleeding episodes, and he was also diagnosed with Hermansky-Pudlak syndrome at 32years of age due to unexplained oculocutaneous albinism. His factor IX level was markedly reduced at 13%; whole exome and Sanger sequencing showed the Durham mutation in F9 (NM_000133.3). The diagnosis of Hermansky-Pudlak syndrome subtype 6 was established by demonstrating absence of platelet delta granules on whole mount electron microscopy, an abnormal secondary wave in platelet aggregation studies, and a novel homozygous c.1114 C>T (p.Arg372*) mutation in HPS6 (NM_024747.5) on exome analysis and Sanger sequencing. Clinical phenotyping revealed no evidence of recurrent or unusual infections, interstitial lung disease or pulmonary fibrosis, or neurological disorders. The patient was treated with fresh frozen plasma, recombinant factor IX, and aminocaproic acid. Treatment with desmopressin was added to his regimen after he was diagnosed with Hermansky-Pudlak syndrome. Treatment of bleeding episodes results in effective hemostasis, and the patient has not required platelet or blood product transfusions. Conclusions This report highlights the need to consider Hermansky-Pudlak syndrome as an etiology of oculocutaneous albinism even in patients with known hematologic disorders associated with bleeding. Identification of a novel mutation in HPS6 in an individual with hemophilia B shows that, although quite rare, patients may be diagnosed with two independent inherited bleeding disorders. No evidence of lung disease was found in this adult patient with Hermansky-Pudlak syndrome subtype 6.

This work describes a 3D, paper-based assay that can isolate sub-populations of cells based on their invasiveness (i.e., distance migrated in a hydrogel) in a gradient of concentration of oxygen (O2). Layers of paper impregnated with a cell-compatible hydrogel are stacked and placed in a plastic holder to form the invasion assay. In most assays, the stack comprises a single layer of paper containing mammalian cells suspended in a hydrogel, sandwiched between multiple layers of paper containing only hydrogel. Cells in the stack consume and produce small molecules; these molecules diffuse throughout the stack to generate gradients in the stack, and between the stack and the bulk culture medium. Placing the cell-containing layer in different positions of the stack, or modifying the permeability of the holder to oxygen or proteins, alters the profile of the gradients within the stack. Physically separating the layers after culture isolates sub-populations of cells that migrated different distances, and enables their subsequent analysis or culture. Using this system, three independent cell lines derived from A549 cancer cells are shown to produce distinguishable migration behavior in a gradient of oxygen. This result is the first experimental demonstration that oxygen acts as a chemoattractant for cancer cells.

Rissone et al. demonstrate that adenylate kinase AK2, an enzyme mutated in reticular dysgenesis (RD) in humans, prevents oxidative stress during hematopoiesis. Using a zebrafish model, as well as induced pluripotent stem cells derived from an RD patient, they find that AK2 deficiency affects hematopoietic stem and progenitor development with increased oxidative stress. Antioxidant treatment rescues the hematopoietic phenotypes., Adenylate kinases (AKs) are phosphotransferases that regulate the cellular adenine nucleotide composition and play a critical role in the energy homeostasis of all tissues. The AK2 isoenzyme is expressed in the mitochondrial intermembrane space and is mutated in reticular dysgenesis (RD), a rare form of severe combined immunodeficiency (SCID) in humans. RD is characterized by a maturation arrest in the myeloid and lymphoid lineages, leading to early onset, recurrent, and overwhelming infections. To gain insight into the pathophysiology of RD, we studied the effects of AK2 deficiency using the zebrafish model and induced pluripotent stem cells (iPSCs) derived from fibroblasts of an RD patient. In zebrafish, Ak2 deficiency affected hematopoietic stem and progenitor cell (HSPC) development with increased oxidative stress and apoptosis. AK2-deficient iPSCs recapitulated the characteristic myeloid maturation arrest at the promyelocyte stage and demonstrated an increased AMP/ADP ratio, indicative of an energy-depleted adenine nucleotide profile. Antioxidant treatment rescued the hematopoietic phenotypes in vivo in ak2 mutant zebrafish and restored differentiation of AK2-deficient iPSCs into mature granulocytes. Our results link hematopoietic cell fate in AK2 deficiency to cellular energy depletion and increased oxidative stress. This points to the potential use of antioxidants as a supportive therapeutic modality for patients with RD.

Smith-Magenis syndrome (SMS), a neurodevelopmental disorder characterized by dysmorphic features, intellectual disability (ID), and sleep disturbances, results from a 17p11.2 microdeletion or a mutation in the RAI1 gene. We performed exome sequencing on 6 patients with SMS-like phenotypes but without chromosomal abnormalities or RAI1 variants. We identified pathogenic de novo variants in two cases, a nonsense variant in IQSEC2 and a missense variant in the SAND domain of DEAF1, and candidate de novo missense variants in an additional two cases. One candidate variant was located in an alpha helix of Necdin (NDN), phased to the paternally inherited allele. NDN is maternally imprinted within the 15q11.2 Prader-Willi Syndrome (PWS) region. This can help clarify NDN's role in the PWS phenotype. No definitive pathogenic gene variants were detected in the remaining SMS-like cases, but we report our findings for future comparison. This study provides information about the inheritance pattern and recurrence risk for patients with identified variants and demonstrates clinical and genetic overlap of neurodevelopmental disorders. Identification and characterization of ID-related genes that assist in development of common developmental pathways and/or gene-networks, may inform disease mechanism and treatment strategies.

Anti-nutritive components in multi resistant potato cultivars were investigated in relation to conventional and organic farming for three years. Glycoalkaloids, nitrate, nitrite, asparagine, and glutamine contents of tubers were examined. Farming technology was found not to have an effect on the level of glycoalkaloids, which was influenced mostly by the genotype and season. Nitrogen fertilisation caused significant increase in nitrate, asparagine, and glutamine contents as compared to organic farming. Nitrite content was found to be more independent of farming technologies than nitrate. Tubers of cultivar Rioja had the lowest nitrate content irrespective of season or technology. In conclusion, the absolute amount and changes of different anti-nutritive components of potato tubers were influenced differently by the technology, genotype, and season in a complex manner. Organic farming had no effect on the glycoalkaloid content, but the nitrate levels had a tendency to be lower compared to conventional farm...

Absent T lymphocytes were unexpectedly found in homozygotes of a transgenic mouse from an unrelated project. T cell development did not progress beyond double-negative stage 1 thymocytes, resulting in a hypocellular, vestigial thymus. B cells were present, but NK cell number and B cell isotype switching were reduced. Transplantation of wild-type hematopoietic cells corrected the defect, which was traced to a deletion involving five contiguous genes at the transgene insertion site on chromosome 12C3. Complementation using bacterial artificial chromosome transgenesis implicated zinc finger BTB-POZ domain protein 1 (Zbtb1) in the immunodeficiency, confirming its role in T cell development and suggesting involvement in B and NK cell differentiation. Targeted disruption of Zbtb1 recapitulated the T−B+NK− SCID phenotype of the original transgenic animal. Knockouts for Zbtb1 had expanded populations of bone marrow hematopoietic stem cells and also multipotent and early lymphoid lineages, suggesting a differentiation bottleneck for common lymphoid progenitors. Expression of mRNA encoding Zbtb1, a predicted transcription repressor, was greatest in hematopoietic stem cells, thymocytes, and pre-B cells, highlighting its essential role in lymphoid development.

CONTEXTUALIZAÇÃO: A Doença Pulmonar Obstrutiva Crônica (DPOC) é uma doença progressiva que reduz a capacidade funcional, comprometendo as atividades de vida diária (AVD). A morbidade e a mortalidade na DPOC apresentam uma estreita relação com a limitação funcional nesses pacientes. OBJETIVOS: Verificar se há associação entre a limitação nas AVD e o índice preditor de mortalidade BODE em pacientes com DPOC moderada a muito grave e não moderada e grave. MÉTODOS: Trinta e nove pacientes com DPOC GOLD 2 a 4, em uma amostra de conveniência, foram submetidos às avaliações: espirometria, índice de massa corpórea (IMC), escala London Chest Activity of Daily Living (LCADL), teste da caminhada de seis minutos (TC6min); escala Medical Research Council (MRC), e o índice BODE foi calculado. O escore total e o percentual do escore total da LCADL (LCADL%total) foram comparados entre os pacientes dos quatro quartis do BODE utilizando-se ANOVA. O Coeficiente de Correlação de Spearman foi utilizado para verificar a associação entre os escores da LCADL e do índice BODE. RESULTADOS: Os pacientes apresentaram em média VEF1%prev=37,0±12,2%, idade=66±8 anos. A LCADL%total apresentou correlação com o BODE (r=0,65, pBACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is a progressive disease that reduces functional capacity, deteriorating the ability to perform activities of daily living (ADL). A close relationship between morbidity and mortality with functional limitation is observed in patients with COPD. OBJECTIVES: To determine if there is a relationship between ADL limitation and the BODE index, which is a predictor of mortality, in patients with moderate to severe COPD. METHODS: Thirty-nine patients with COPD GOLD 2 to 4 recruited by convenience, were submitted to the following tests: spirometry, body mass index (BMI), the London Chest Activity of Daily Living (LCADL) scale, six-minute walking test (6MWT), the Medical Research Council (MRC) scale and the BODE index was calculated. The total score and the percentage of the total score LCADL (LCADL%total) were compared between patients of the four quartiles of the BODE using the Analysis of Variance test. The Spearman correlation coefficient was used to investigate the association between scores of LCADL and BODE index. RESULTS: Patients had an average of FEV1%pred=37±12% and were on average 66±8 years-old. The LCADL%total correlated with the BODE index (r=0.65, p

CONTEXTUALIZAÇÃO: A Doença Pulmonar Obstrutiva Crônica (DPOC) é uma doença progressiva que reduz a capacidade funcional, comprometendo as atividades de vida diária (AVD). A morbidade e a mortalidade na DPOC apresentam uma estreita relação com a limitação funcional nesses pacientes. OBJETIVOS: Verificar se há associação entre a limitação nas AVD e o índice preditor de mortalidade BODE em pacientes com DPOC moderada a muito grave e não moderada e grave. MÉTODOS: Trinta e nove pacientes com DPOC GOLD 2 a 4, em uma amostra de conveniência, foram submetidos às avaliações: espirometria, índice de massa corpórea (IMC), escala London Chest Activity of Daily Living (LCADL), teste da caminhada de seis minutos (TC6min); escala Medical Research Council (MRC), e o índice BODE foi calculado. O escore total e o percentual do escore total da LCADL (LCADL%total) foram comparados entre os pacientes dos quatro quartis do BODE utilizando-se ANOVA. O Coeficiente de Correlação de Spearman foi utilizado para verificar a associação entre os escores da LCADL e do índice BODE. RESULTADOS: Os pacientes apresentaram em média VEF1%prev=37,0±12,2%, idade=66±8 anos. A LCADL%total apresentou correlação com o BODE (r=0,65, p

INTRODUÇÃO E OBJETIVO: A DPOC é importante causa de mortalidade no mundo. Para predizer a mortalidade em pacientes com doença pulmonar obstrutiva crônica (DPOC), o índice BODE é utilizado e sua pontuação pode ser reduzida com a prática de atividade física. Este estudo objetiva verificar a associação entre o índice BODE com atividade física em indivíduos com DPOC. MÉTODOS: Foi calculado o índice BODE usando as variáveis: distância no teste da caminhada de seis minutos (TC6'); Medical Research Council Dyspnea Score (MRC); índice de massa corpórea (IMC) e grau de obstrução ao fluxo aéreo (VEF1). Os indivíduos foram divididos em grupos classificados em ativos fisicamente (AF) e inativos fisicamente (IF). O teste do qui-quadrado foi utilizado para testar a independência das variáveis. O teste t foi utilizado para comparar as variáveis TC6', MRC, IMC, VEF1 entre os grupos de indivíduos AF e IF. RESULTADOS E CONCLUSÃO: Em 38 pacientes com moderada e grave obstrução ao fluxo aéreo (36,5 ± 11,9), o valor médio do BODE foi de 4,1 ± 2,1 para o grupo de indivíduos AF; para o grupo de indivíduos IF, o valor médio do BODE foi de 6 ± 2,2. Houve diferença significativa na comparação das variáveis TC6', MRC, BODE entre os grupos. Existiu maior ocorrência de indivíduos que praticam atividade física de se situar nos quartis um e dois e os indivíduos que não praticam atividade física se situarem nos quartis três e quatro. A existência da maior pontuação do BODE para os pacientes com DPOC considerados inativos fisicamente sustenta o conceito geral de que a inatividade aumenta o risco de morte.INTRODUCTION: COPD is an important cause of morbidity and mortality in the world. The BODE is an important predictive index of mortality and its scores can be reduced with the practice of physical activity. OBJECTIVE: To verify the association between BODE and physical activity in patients with chronic obstructive pulmonary disease (COPD). METHODS: the BODE index was calculated using the following variables: distance in the 6-minute walk test (6MWT); Medical Research Council Dyspnea Score (MRC); body mass index (BMI) and degree of pulmonary function (FEV1). The patients were divided in groups classified as physically active (PA) or physically inactive (PI). Qui-square test was used to verify the independence of the variables. T-Student test was used to compare the 6WMT, MRC, BMI, FEV1 variables between PA and PI groups. RESULTS AND CONCLUSION: In 38 patients with moderate and serious obstruction to the air flow (36.5 ± 11.9), BODE mean value was of 4.1/ ±/ 2.1 for the PA group, while for the PI group BODE mean value was of ± 2.2. Significant difference in the comparison of WD6', MRC, BODE variables between groups was observed. There was more occurrence of individuals who practice physical activity in quartiles one and two, while individuals who do not practice aerobic physical activity were in quartiles three and four. Higher scores of BODE for DPOC patients considered physically inactive supports the general concept that inactivity increases risk of death.

Chemical gradients play an important role in guiding the activities of both eukaryotic and prokaryotic cells. Here, we used molecularly well-defined chemical gradients formed by self-assembled monolayers (SAMs) on gold films to reveal that mammalian cell adhesion and bacterial biofilm formation respond differently to a gradient of surface chemistry that resists cell attachment. Gradient self-assembled monolayers (SAMs) consisting of two mixed alkanethiols were fabricated by differential exposure of the gold film to one alkanethiol, followed by soaking in another alkanethiol solution. A gradient in bioinertness that resisted cell attachment was created on SAMs from a gradient in the surface density of HS(CH2)11(OCH2CH2)3OH, backfilled with either HS(CH2)11OH or HS(CH2)11CH3. Measurements of the amounts of mammalian cells and bacterial biofilms on these gradient surfaces reveal that, for mammalian cells, a critical density of adhesion ligands from absorbed proteins on surfaces exists for supporting maximum adhesion and proliferation, whereas for the bacterium Escherichia coli , the amount of biofilm formed on surfaces increased linearly with the surface density of adhesive groups (methyl or hydroxyl groups) in different media. These results are consistent with mammalian cell adhesion requiring an anchorage via specific molecular recognitions and suggest that biofilms can form by immobilization of bacteria via nonspecific interaction between bacteria and surfaces.

Human adenylate kinase 2 deficiency causes the most profound human haematopoietic defect associated with sensorineural deafness. Reticular dysgenesis (RD) is an autosomal recessive form of human severe combined immunodeficiency (SCID) characterized by an early differentiation block in the myeloid lineage and a profound impairment in lymphoid maturation associated with bilateral sensorineural deafness. The lack of polymorphonuclear neutrophils in affected newborns is responsible for the occurrence of severe infections earlier than usually observed in the other forms of SCID. Furthermore, RD associated neutropenia is characterized by the lack of responsiveness to G-CSF. We have identified bi-allelic mutations in the adenylate kinase 2 (AK2) gene is seven patients affected with RD. These mutations resulted in the absence or a strong decrease in protein expression. Restoration of AK2 expression in the bone marrow cells of RD patients overcomes the neutrophil differentiation arrest, underlying its specific requirement in the development of a restricted set of haematopoietic lineages. Lastly, we established that AK2 is specifically expressed in the stria vascularis region of the inner ear. The function of this gene in the differentiation of a given set of cell lineages is rapidly in progress, showing that studies of primary immunodeficiencies continue to provide key information on human lympho-haematopoietic development. Moreover the AK2 enzyme seems a key molecule in different biological systems such as the lympho-haematopoiesis and the brain development.

A collection of structurally closely related furanones was synthesized to identify the most important structural elements in brominated furanones for inhibiting the formation of bacterial biofilms. The results suggest that a conjugated exocyclic vinyl bromide on the furanone ring is the most important structural element for the non-toxic but inhibition activity for Escherichia coli biofilm formation. Furanones bearing monosubstituted bromide groups on saturated carbons were found to have a toxic effect that attenuates the bacterial growth.

Bacterial biofilms cause serious problems, such as antibiotic resistance and medical device-related infections. To further understand bacterium-surface interactions and to develop efficient control strategies, self-assembled monolayers (SAMs) of alkanethiols presenting different functional groups on gold films were analyzed to determine their resistance to biofilm formation. Escherichia coli was labeled with green florescence protein, and its biofilm formation on SAM-modified surfaces was monitored by confocal laser scanning microscopy. The three-dimensional structures of biofilms were analyzed with the COMSTAT software to obtain information about biofilm thickness and surface coverage. SAMs presenting methyl, l -gulonamide (a sugar alcohol tethered with an amide bond), and tri(ethylene glycol) (TEG) groups were tested. Among these, the TEG-terminated SAM was the most resistant to E. coli biofilm formation; e.g., it repressed biofilm formation by E. coli DH5α by 99.5% ± 0.1% for 1 day compared to the biofilm formation on a bare gold surface. When surfaces were patterned with regions consisting of methyl-terminated SAMs surrounded by TEG-terminated SAMs, E. coli formed biofilms only on methyl-terminated patterns. Addition of TEG as a free molecule to growth medium at concentrations of 0.1 and 1.0% also inhibited biofilm formation, while TEG at concentrations up to 1.5% did not have any noticeable effects on cell growth. The results of this study suggest that the reduction in biofilm formation on surfaces modified with TEG-terminated SAMs is a result of multiple factors, including the solvent structure at the interface, the chemorepellent nature of TEG, and the inhibitory effect of TEG on cell motility.

It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20-37 °C, which can be rapidly converted into a free-flowing dispersion of spheres on cooling to 5 °C. Functionalization of the worms with disulfide groups was found to be essential for ensuring sufficient mechanical stability of these hydrogels to enable long-term cell culture. These disulfide groups are conveniently introduced via statistical copolymerization of a disulfide-based dimethacrylate under conditions that favor intramolecular cyclization and subsequent thiol/disulfide exchange leads to the formation of reversible covalent bonds between adjacent worms within the gel. This new approach enables cells to be embedded within micrometer-thick slabs of gel with good viability, permits cell culture for at least 12 days, and facilitates recovery of viable cells from the gel simply by incubating the culture in buffer at 4 °C (thus, avoiding the enzymatic degradation required for cell harvesting when using commercial protein-based gels, such as Matrigel).

This work demonstrates the application of a 3D culture system-Cells-in-Gels-in-Paper (CiGiP)-in evaluating the metabolic response of lung cancer cells to ionizing radiation. The 3D tissue-like construct-prepared by stacking multiple sheets of paper containing cell-embedded hydrogels-generates a gradient of oxygen and nutrients that decreases monotonically in the stack. Separating the layers of the stack after exposure enabled analysis of the cellular response to radiation as a function of oxygen and nutrient availability; this availability is dictated by the distance between the cells and the source of oxygenated medium. As the distance between the cells and source of oxygenated media increased, cells show increased levels of hypoxia-inducible factor 1-alpha, decreased proliferation, and reduced sensitivity to ionizing radiation. Each of these cellular responses are characteristic of cancer cells observed in solid tumors. With this setup we were able to differentiate three isogenic variants of A549 cells based on their metabolic radiosensitivity; these three variants have known differences in their metastatic behavior in vivo. This system can, therefore, capture some aspects of radiosensitivity of populations of cancer cells related to mass-transport phenomenon, carry out systematic studies of radiation response in vitro that decouple effects from migration and proliferation of cells, and regulate the exposure of oxygen to subpopulations of cells in a tissue-like construct either before or after irradiation.

This work reports a new biocompatible surfactant structure, of which the hydrophilic head group is composed of a folded, stable self-inclusion complex of a ferrocenyl substituted beta-cyclodextrin (betaCD). While multiple intra- or intermolecular complexes can exist for this amphiphile, the molecule folds into a unique intramolecular complex with well-defined conformation, in which part of the aliphatic chain and the ferrocene group are both included in the annular cavity of betaCD. Study of different isosteric covalent linkages indicates that this folded structure is stable against displacement by the presence of other small guest molecules. Furthermore, in contrast to ferrocene-CD conjugates that are without the aliphatic chain, the presence of small guest molecules in solution does not influence at all the induced circular dichroism signal of this amphiphile, indicating a sterically congested, but stable, folded conformation of the inclusion complex. This new amphiphile is surface active and, more importantly, does not denature the membrane protein bacteriorhodopsin. Finally, because this surfactant forms self-assembled aggregates, this work introduces a folded structure into soft matters formed by amphiphiles in water.

The Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency characterized by recurrent infections, thrombocytopenia, eczema, and high incidence of malignancy and autoimmunity. The cellular mechanisms underlying autoimmune complications in WAS have been extensively studied; however, they remain incompletely defined. We investigated the characteristics of IL-10-producing CD19+CD1dhighCD5+ B cells (CD1dhighCD5+ Breg) obtained from Was gene knockout (WKO) mice and found that their numbers were significantly lower in these mice compared to wild type (WT) controls. Moreover, we found a significant age-dependent reduction of the percentage of IL-10-expressing cells in WKO CD1dhighCD5+ Breg cells as compared to age-matched WT control mice. CD1dhighCD5+ Breg cells from older WKO mice did not suppress the in vitro production of inflammatory cytokines from activated CD4+ T cells. Interestingly, CD1dhighCD5+ Breg cells from older WKO mice displayed a basal activated phenotype which may prevent normal cellular responses, among which is the expression of IL-10. These defects may contribute to the susceptibility to autoimmunity with age in patients with WAS.

X-linked severe combined immunodeficiency (XSCID) is caused by mutations in the IL-2 receptor gamma chain (IL2RG) gene, resulting in absent T lymphocytes and nonfunctional B lymphocytes. Recently T lymphocyte production and B lymphocyte function were restored in XSCID patients infused with autologous stem cells transduced with a retrovirus containing the human IL2RG cDNA. To optimize the expression of human IL2RG for future clinical trials, we compared five retroviral vectors expressing human IL2RG from different LTR enhancer-promoter elements in a mouse model. Northern and Southern blot analysis of hematopoietic tissues from repopulated mice revealed that the retroviral vector with the highest expression per copy number was MFG-S-hIL2RG, followed by MND-hIL2RG. All five vectors were capable of restoring lymphopoiesis in irradiated XSCID mice transplanted with transduced IL2RG-deficient hematopoietic stem cells. Transduction of IL2RG-deficient hematopoietic stem cells with all five vectors restored T lymphopoiesis in transplanted stem cell-deficient W/W(v) mouse recipients. However, only XSCID stem cells transduced with the MFG-S-hIL2RG vector generated B lymphocytes in W/W(v) mice. We conclude that the MFG-S-hIL2RG vector provides the best opportunity for in vivo selection and development of B and T lymphocytes for human XSCID gene therapy.

Most humans with autoimmune lymphoproliferative syndrome (ALPS) carry heterozygous dominant mutations in one allele of the gene encoding Fas/APO-1/CD95. ALPS patients, like Fas-deficient MRL lpr/lpr mice, have lymphoproliferation, autoimmunity, increased CD4(-)/CD8(-) T lymphocytes, and apoptosis defects. Consistent with the phenotypic variability of lpr/lpr mice of different background strains, human genetic studies indicate that a Fas mutation is insufficient to induce ALPS in all mutation carriers. To investigate the dominant function of human Fas mutations and the additional genetic factor(s) involved in the development of ALPS, we generated transgenic mice expressing, in addition to endogenous Fas, mouse Fas molecules bearing mutations in the intracellular death domain corresponding to mutations identified in ALPS patients. Transgenic mice developed mild features of ALPS, including hepatosplenomegaly, elevated proportions of lymphocytes in spleen and lymph nodes, apoptotic defects, and hepatic lymphocytic infiltrates. Therefore defective murine Fas proteins act in a dominant manner to impair apoptosis of activated lymphocytes and disrupt lymphocyte homeostasis. The influence of genetic background on phenotype was studied by comparing transgenic mice on FVB/N and (FVB/N x MRL) backgrounds with syngenetic control mice and with MRL and MRL lpr/lpr mice. While expression of transgenic mutant Fas contributed mainly to hepatosplenomegaly and accumulation of lymphocytes, MRL background genes played a major role in the production of autoantibodies and elevated serum immunoglobulin levels. Moreover, compared to FVB/N (+/+) mice, a substantial Fas-specific apoptotic defect was found in MRL (+/+) mice, suggesting a mechanism for the known tendency of this strain to develop autoimmunity.

The hemoglobin-deficit mouse mutant ( hbd ) is characterized by a severe microcytic anemia that is inherited in an autosomal-recessive manner. Previous results from our laboratory indicated that normal mice develop anemia if they are transplanted with bone marrow from mutant animals. Furthermore, we demonstrated a delay in erythroid reconstitution from hbd marrow compared to normal marrow. Although these experiments show a defect that is intrinsic to hbd marrow, it is unclear if the hbd phenotype is solely the result of a bone marrow-derived defect. To exclude an environmental influence on hbd anemia, we attempted to cure the defect by transplanting normal marrow into the hbd mouse. We observed that the transplanted animals converted to a normal phenotype. These results indicated that the defect is bone marrow derived. In contrast to the microcytosis mutant whose defective gene is ubiquitiously expressed, our data suggest that the defective gene product is specific to hematopoietic cells.

The mouse mutant hemoglobin deficit (gene symbol hbd ) is characterized by a severe microcytic anemia that is inherited in an autosomal-recessive manner. To assess the mutation's effect on hematopoiesis, unfractionated bone marrow (BM) from either a mutant C57BL6/J-hbd/hbd, Gpi1b/Gpi1b (phenotype symbol HBD), or normal C57BL6/J -+hbd/+hbd, Gpi1b/Gpi1b mouse was injected intravenously into irradiated congenic C57BL6/J-+hbd/+hbd, Gpi1a/Gpi1a, Igha/Igha, Thy1a/Thy1a mice. The congenic recipients of mutant or normal marrow obtained complete red blood cell (RBC) and leukocyte reconstitution, with the exception of one recipient of HBD marrow. After 24 weeks posttransplantation, the normal recipients of HBD marrow obtained a microcytic anemia similar to the donor. These results suggest that the HBD phenotype is caused by a BM defect. We observed that the erythroid lineage derived from donor HBD marrow repopulated more slowly than the normal marrow at 4 weeks posttransplantation. To determine if this difference was a result of an erythropoietic defect, competitive repopulation was performed using either mutant or normal marrow competed against normal congenic marrow. For the erythroid lineage, no significant contribution from HBD marrow was observed. To assess if the RBC block was based on a deficiency of myeloid progenitors, both in vitro and in vivo assays were performed: absolute numbers of bone progenitors were increased, suggesting that the defect results in a late block to erythroid differentiation.

Three-dimensional (3D) culture systems can mimic certain aspects of the cellular microenvironment found in vivo, but generation, analysis and imaging of current model systems for 3D cellular constructs and tissues remain challenging. This work demonstrates a 3D culture system – Cells-in-Gels-in-Mesh (CiGiM) – that uses stacked sheets of polymer-based mesh to support cells embedded in gels to form tissue-like constructs; the stacked sheets can be disassembled by peeling the sheets apart to analyze cultured cells—layer-by-layer—within the construct. The mesh sheets leave openings large enough for light to pass through with minimal scattering, and thus allowing multiple options for analysis—(i) using straightforward analysis by optical light microscopy, (ii) by high-resolution analysis with fluorescence microscopy, or (iii) with a fluorescence gel scanner. The sheets can be patterned into separate zones with paraffin film-based decals, in order to conduct multiple experiments in parallel; the paraffin-based decal films also block lateral diffusion of oxygen effectively. CiGiM simplifies the generation and analysis of 3D culture without compromising throughput, and quality of the data collected: it is especially useful in experiments that require control of oxygen levels, and isolation of adjacent wells in a multi-zone format.

Background: In addition to NBEAL2, a single dominant-negative mutation in the GFIB1 gene has been associated with grey platelet syndrome but no association has been mentioned with dense granule deficiency. Here we describe a child with thrombocytopenia, alpha-delta granule deficiency, and a homozygous missense mutation in GFIB1. Case Report:The patient is an 8y/o Hispanic male born to non-consanguineous parents. Prenatal and birth history were unremarkable. No family history of blood disorders or pediatric malignancies. The boy has 2 healthy older siblings, no dysmorphic features, and normal skin pigmentation and eye findings. Renal function and hearing are normal. At 3mo he was noted to have multiple spontaneous petechiae along with an isolated thrombocytopenia of 46K/uL. At 10 months, a bone marrow evaluation showed increased megakaryocytes suggestive of ITP. By 3years of age, he received three treatments of IVIG without an adequate response. His platelet counts have generally ranged between 30-50K/uL. With acute illnesses, they drop to 15-20K/uL. His bleeding symptoms have primarily been spontaneous bruising and petechiae as well as prolonged epistaxis. His symptoms have been generally controlled with anti-fibrinolytic agents alone. Platelet transfusions have been reserved for surgical procedures or significant bleeding symptoms. Further evaluations over the past 5 years have included peripheral smears, showing atypical large hypo-granular platelets, and two additional bone marrow aspirates, showing megakaryocytic hyperplasia with numerous osteoclast-like forms and occasional small mono-lobated megakaryocytes and evident emperipolesis. Anti-nuclear antibody testing and platelet direct and indirect antibody testing were negative. Platelet electron microscopy showed that platelets virtually contained no dense granules (0.05 dense granules/platelet, 200 platelets) and about 30-40% of the platelets had markedly decreased alpha granules. Some platelets had complex canalicular networks and membrane vacuoles. These features are consistent with an alpha-delta platelet storage-pool deficiency. A myeloid malignancy mutation panel, performed using next generation sequencing, detected no variants of known significance. Gene sequencing for MHY9-related disorders as well as DNA breakage analysis for Fanconi's anemia were negative. However, DNA sequencing of the patient's sample revealed a homozygous missense mutation (c923 T>C; p.Leu308Pro) in the GFIB1gene; each parent carried one copy of this change. Conclusion: GFIB1, mapped to chromosome 9q34.13, encodes a transcriptional repressor that is important for megakaryopoiesis. It has been reported in patients with gray platelet syndrome; an inherited platelet disorder associated with thrombocytopenia and decreased alpha granules. This case points to an association of biallelic GFIB1 mutations with alpha-delta granule deficiency. This case underscores the importance of platelet esoteric testing and molecular analysis in the diagnosis of hereditary platelet disorders. As a targetable mutation in the future, this could revolutionize the early diagnosis and treatment of this rare platelet disorder. Disclosures No relevant conflicts of interest to declare.

This paper describes a programme for inpatients with severe personality disorders as well as acute exacerbation of Axis II pathology. The goal of the programme was to quickly stabilize participants with respect to self-injurious behaviour and return them to the community. The programme consisted of frequent coordinated cognitive group therapy sessions with supporting written and audiovisual materials. The groups were open-ended and heterogenous, and employed educational and experiential-affective components. An extended group case example is presented in order to illustrate the operation of the programme. The range of typical participants is described, including their diagnoses, treatment plans, and homework assignments. Some comments on the personal and professional difficulties encountered by therapists working with this population are described as well as some suggestions for alleviating those difficulties.

Crystallization of proteins is important for fundamental studies and biopharmaceutical development but remains largely an empirical science. Here, we report the use of organic salts that can form a class of unusual non-amphiphilic lyotropic liquid crystals to crystallize the protein lysozyme. Certain non-amphiphilic organic molecules with fused aromatic rings and two charges can assemble into stable thread-like noncovalent polymers that may further form liquid crystal phases in water, traditionally termed chromonic liquid crystals. Using five of these mesogenic molecules as additives to induce protein crystallization, we discover that molecules that can form liquid crystal phases in water are highly effective at inducing the crystal formation of lysozyme, even at concentrations significantly lower than that required for forming liquid crystal phases. This result reveals an example of inducing protein crystallization by the molecular assembly of the additives, and is consistent with a new mechanism by which the strong hydration of an assembly process provides a gradual means to compete for the water molecules to enable solvated proteins to form crystals.

This work studies the phase separations between polymers and a small molecule in a common aqueous solution that do not have well-defined hydrophobic-hydrophilic separation. In addition to poly(acrylamide) (PAAm) and poly(vinyl alcohol) (PVA), poly(vinyl pyrrolidone) (PVP) also promotes liquid crystal (LC) droplet formation by disodium cromoglycate (5'DSCG) solvated in water. In the presence of these polymers, the concentration of 5'DSCG needed for forming LC droplets is substantially lower than that needed for forming an LC phase by 5'DSCG alone. To define the concentration ranges that 5'DSCG molecules form liquid crystals (either as droplets or as an isotropic-LC mixture), we constructed ternary phase diagrams for 5'DSCG, water, and a polymer - PVA, PVP, or PAAm. We discovered that PAAm with high molecular weight promotes LC droplet formation by 5'DSCG more effectively than PAAm with low molecular weight. At the same weight percentage, long-chain PAAm can cause 5'DSCG to form LC droplets in water, whereas short-chain PAAm does not. Poly(vinyl pyrrolidone) (PVP), which has functional groups that are more dissimilar to 5'DSCG than PVA and PAAm, promotes LC droplet formation by 5'DSCG more effectively than either of the other two polymers. Additionally, small angle neutron scattering data revealed that the assembly structure of 5'DSCG promoted by the presence of PVA is similar to the thread structure formed by 5'DSCG alone. Together, these results reveal how noncovalent polymerization can be promoted by mixing thermodynamically incompatible molecules and elucidate the basic knowledge of nonamphiphilic colloidal science.

Chronic Obstructive Pulmonary Disease (COPD) is a progressive disease that reduces functional capacity, deteriorating the ability to perform activities of daily living (ADL). A close relationship between morbidity and mortality with functional limitation is observed in patients with COPD.To determine if there is a relationship between ADL limitation and the BODE index, which is a predictor of mortality, in patients with moderate to severe COPD.Thirty-nine patients with COPD GOLD 2 to 4 recruited by convenience, were submitted to the following tests: spirometry, body mass index (BMI), the London Chest Activity of Daily Living (LCADL) scale, six-minute walking test (6MWT), the Medical Research Council (MRC) scale and the BODE index was calculated. The total score and the percentage of the total score LCADL (LCADL%total) were compared between patients of the four quartiles of the BODE using the Analysis of Variance test. The Spearman correlation coefficient was used to investigate the association between scores of LCADL and BODE index.Patients had an average of FEV1%pred=37±12% and were on average 66±8 years-old. The LCADL%total correlated with the BODE index (r=0.65, p0.05) as well as with the variables FEV1, dyspnea and walked distance in the 6MWT (r=-0.42, r=0.76 and r=-0.67, p0.05, respectively). The comparison of the average scores of the LCADL%total between BODE quartiles 1, 2, 3 and 4, demonstrated that only the 4th quartile differed significantly from the others (p0.05).ADL limitation has a strong association with the BODE index in patients with moderate to severe COPD and with three of the four variables that composes it.

Self-assembly of large quantities of entirely water-soluble molecules is entropically challenging. In this work, we describe the design and synthesis of water-soluble aromatic (dichromonyl) molecules that can form nonamphiphilic assemblies and the so-called chromonic liquid crystal phase in water. We discover a new molecule, 5'DSCG-diviol, that exhibits a large birefringent phase, and we show that the formation of this unique class of nonamphiphilic lyotropic liquid crystal shares enormous similarity to the polymorphism observed for crystal formation. Small-angle neutron scattering (SANS) revealed a concentration-independent rod-shaped assembly at concentrations below and above the formation of liquid crystal phase. Adding a small percentage of monoanionic aromatic molecules to the liquid crystal resulted in the elimination of the liquid crystal phase, but addition of dianionic aromatic molecules retained the liquid crystal phase. Together, these results suggest a new assembly structure for nonamphiphilic molecules in water, which is comprised of long threads of small molecules connected by salt bridges stacked over aromatic groups, with the molecular threads heavily hydrated with solvent water. Furthermore, mixing molecules with different structures can result in new liquid crystalline materials, or in segregation of the molecules into different solvation volumes, each of which contains only one type of molecule. The unusual thermodynamic incompatibility of entirely water-soluble molecules also supports the model of molecular threads, in which two polymer-like assemblies do not mix.

INTRODUÇÃO E OBJETIVO: A DPOC é importante causa de mortalidade no mundo. Para predizer a mortalidade em pacientes com doença pulmonar obstrutiva crônica (DPOC), o índice BODE é utilizado e sua pontuação pode ser reduzida com a prática de atividade física. Este estudo objetiva verificar a associação entre o índice BODE com atividade física em indivíduos com DPOC. MÉTODOS: Foi calculado o índice BODE usando as variáveis: distância no teste da caminhada de seis minutos (TC6'); Medical Research Council Dyspnea Score (MRC); índice de massa corpórea (IMC) e grau de obstrução ao fluxo aéreo (VEF1). Os indivíduos foram divididos em grupos classificados em ativos fisicamente (AF) e inativos fisicamente (IF). O teste do qui-quadrado foi utilizado para testar a independência das variáveis. O teste t foi utilizado para comparar as variáveis TC6', MRC, IMC, VEF1 entre os grupos de indivíduos AF e IF. RESULTADOS E CONCLUSÃO: Em 38 pacientes com moderada e grave obstrução ao fluxo aéreo (36,5 ± 11,9), o valor médio do BODE foi de 4,1 ± 2,1 para o grupo de indivíduos AF; para o grupo de indivíduos IF, o valor médio do BODE foi de 6 ± 2,2. Houve diferença significativa na comparação das variáveis TC6', MRC, BODE entre os grupos. Existiu maior ocorrência de indivíduos que praticam atividade física de se situar nos quartis um e dois e os indivíduos que não praticam atividade física se situarem nos quartis três e quatro. A existência da maior pontuação do BODE para os pacientes com DPOC considerados inativos fisicamente sustenta o conceito geral de que a inatividade aumenta o risco de morte. INTRODUCTION: COPD is an important cause of morbidity and mortality in the world. The BODE is an important predictive index of mortality and its scores can be reduced with the practice of physical activity. OBJECTIVE: To verify the association between BODE and physical activity in patients with chronic obstructive pulmonary disease (COPD). METHODS: the BODE index was calculated using the following variables: distance in the 6-minute walk test (6MWT); Medical Research Council Dyspnea Score (MRC); body mass index (BMI) and degree of pulmonary function (FEV1). The patients were divided in groups classified as physically active (PA) or physically inactive (PI). Qui-square test was used to verify the independence of the variables. T-Student test was used to compare the 6WMT, MRC, BMI, FEV1 variables between PA and PI groups. RESULTS AND CONCLUSION: In 38 patients with moderate and serious obstruction to the air flow (36.5 ± 11.9), BODE mean value was of 4.1/ ±/ 2.1 for the PA group, while for the PI group BODE mean value was of ± 2.2. Significant difference in the comparison of WD6', MRC, BODE variables between groups was observed. There was more occurrence of individuals who practice physical activity in quartiles one and two, while individuals who do not practice aerobic physical activity were in quartiles three and four. Higher scores of BODE for DPOC patients considered physically inactive supports the general concept that inactivity increases risk of death.

OBJETIVO: Traduzir a escala London Chest Activity of Daily Living (LCADL) para o português e verificar se essa versão é reprodutível em pacientes com doença pulmonar obstrutiva crônica (DPOC) grave no Brasil. MÉTODOS: Foram realizadas a tradução da escala LCADL para o português e a tradução retrógrada dessa versão em português para o inglês. Essa primeira versão em português foi aplicada a 8 pacientes com DPOC, e possíveis dificuldades em relação ao texto foram investigadas. As principais dificuldades encontradas foram discutidas com os autores da escala, chegando-se a uma versão final do instrumento. Essa versão final foi aplicada duas vezes a 31 pacientes com DPOC por dois observadores separadamente em um primeiro dia. Após 15-20 dias, essa mesma versão foi aplicada novamente aos mesmos pacientes por um dos observadores. No primeiro dia os pacientes foram submetidos à prova de função pulmonar e ao teste de caminhada de seis minutos (TC6). RESULTADOS: A versão brasileira da escala LCADL demonstrou excelente reprodutibilidade no escore total e na maioria das questões, com um coeficiente alfa de Cronbach interobservador de 0,97 (IC95%: 0,89-0,97; p < 0,05) e um coeficiente alfa de Cronbach intra-observador de 0,96 (IC95%: 0,83-0,96; p < 0,05). O escore total dessa versão apresentou correlação negativa com o volume expiratório forçado no primeiro segundo em litros (r = -0,49; p < 0,05) e a distância percorrida no TC6 (r = -0,56; p < 0,05). CONCLUSÃO: A versão brasileira da escala LCADL é um instrumento confiável, reprodutível e válido para avaliar a dispnéia durante atividades de vida diária em pacientes com DPOC grave. OBJECTIVE: To translate the London Chest Activity of Daily Living (LCADL) scale into Portuguese and to determine whether this version is reproducible in Brazilian patients with severe chronic obstructive pulmonary disease (COPD). METHODS: The LCADL scale was translated into Portuguese and then back-translated into English. This pilot Brazilian Portuguese version was administered to 8 patients with COPD, and possible text-related problems were investigated. The principal problems were discussed with the authors of the original scale, and a final translated version was arrived at. At the study outset, two observers administered this final version (twice in one day) to 31 patients with COPD. One of those observers again administered the scale to the same patients 15-20 days later. At baseline, the patients were submitted to pulmonary function testing and to the six-minute walk test (6MWT). RESULTS: The Brazilian Portuguese version of the LCADL scale demonstrated excellent reproducibility in the total score and in most of the questions, with an inter-rater Cronbach's alpha coefficient of 0.97 (95% CI: 0.89-0.97; p < 0.01) and an intra-rater Cronbach's alpha coefficient of 0.96 (95% CI: 0.83-0.96; p < 0.01). The total score presented a negative correlation with forced expiratory volume in one second in liters (r = -0.49; p < 0.05) and with distance covered on the 6MWT (r = -0.56; p < 0.05). CONCLUSION: The Brazilian Portuguese version of the LCADL scale is a reliable, reproducible, and valid instrument for evaluating dyspnea during activities of daily living in patients with severe COPD.

This clinical outcome study compared four separate cognitive behavioral and one behavioral treatment for reducing social anxiety with a waiting list control. Subjects were adults who responded to advertisements for treatment. In the cognitive-behavioral conditions, subjects were trained to modify their disruptive cognitions and/or emit adaptive cognitions relevant to interpersonal behavior with the specific content and techniques determined by the particular cognitive therapy. In the behavioral condition, subjects modeled and rehearsed appropriate social interpersonal behaviors. Subjects were assessed pre and post-treatment on self-report measures of social anxiety, measures of general anxiety, clinical scales of anxiety, depression and hostility, and behavioral, and physiological concomitants of social anxiety. Subjects in all active treatment conditions showed improvement on the measures of social anxiety and the behavioral measure. However, only those in the cognitive treatment conditions, demonstrated treatment effects on general anxiety and the clinical scales. The results indicate the efficacy of both cognitive and behavioral approaches to the reduction of social anxiety. However, they fail to demonstrate clear distinctions between the various cognitive therapies.

This work reports the design and use of a gradient nanotopography to enhance the resistance of mammalian cell adhesion on surfaces. The gradient nanotopography is prepared by depositing gold films on a glass microscope slide in a single azimuthal direction, but with continuous increase in the angle between the incidence of gold atoms and the normal of the surface. By forming patterned self-assembled monolayers (SAMs) of alkanethiols into cell-adhesive regions (HS(CH2)14CH3) surrounded by cell-resistant areas (HS(CH2)12(OCH2CH2)3OH) on the gold film, the adhesion and confinement of albino 3T3 fibroblasts along the topographic gradient are studied. At the early stage of cell culture, cells adhere faster and reach confluency sooner on higher topographic regions (gold deposited at large incident angle) than on lower topographic regions (gold deposited at small incident angle). After cells proliferate to reach confluency within the patterns, the cells are confined in the high topographic regions up to 25 days,...

To translate the London Chest Activity of Daily Living (LCADL) scale into Portuguese and to determine whether this version is reproducible in Brazilian patients with severe chronic obstructive pulmonary disease (COPD).The LCADL scale was translated into Portuguese and then back-translated into English. This pilot Brazilian Portuguese version was administered to 8 patients with COPD, and possible text-related problems were investigated. The principal problems were discussed with the authors of the original scale, and a final translated version was arrived at. At the study outset, two observers administered this final version (twice in one day) to 31 patients with COPD. One of those observers again administered the scale to the same patients 15-20 days later. At baseline, the patients were submitted to pulmonary function testing and to the six-minute walk test (6MWT).The Brazilian Portuguese version of the LCADL scale demonstrated excellent reproducibility in the total score and in most of the questions, with an inter-rater Cronbach's alpha coefficient of 0.97 (95% CI: 0.89-0.97; p0.01) and an intra-rater Cronbach's alpha coefficient of 0.96 (95% CI: 0.83-0.96; p0.01). The total score presented a negative correlation with forced expiratory volume in one second in liters (r = -0.49; p0.05) and with distance covered on the 6MWT (r = -0.56; p0.05).The Brazilian Portuguese version of the LCADL scale is a reliable, reproducible, and valid instrument for evaluating dyspnea during activities of daily living in patients with severe COPD.

Emulsion systems involving surfactants are mainly driven by the separation of the hydrophobic interactions of the aliphatic chains from the hydrophilic interactions of amphiphilic molecules in water. In this study, we report an emulsion system that does not include amphiphilic molecules but molecules with functional groups that are completely solvated in water. These functional groups give rise to molecular interactions including hydrogen bonding, pi stacking, and salt bridging and are segregated into a dispersion of droplets forming a water-in-water emulsion. This water-in-water emulsion consists of dispersing droplets of a water-solvated biocompatible liquid crystal--disodium cromoglycate (DSCG)--in a continuous aqueous solution containing specific classes of water-soluble polymers. Whereas aqueous solutions of polyols support the formation of emulsions of spherical droplets consisting of lyotropic liquid crystal DSCG with long-term stability (for at least 30 days), aqueous solutions of polyamides afford droplets of DSCG in the shape of prolate ellipsoids that are stable for only 2 days. The DSCG liquid crystal in spherical droplets assumes a radial configuration in which the optical axis of the liquid crystal aligns perpendicular to the surface of the droplets but assumes a tangential configuration in prolate ellipsoids in which the optical axis of the liquid crystal aligns parallel to the surface of the droplet. Other classes of water-soluble polymers including polyethers, polycations, and polyanions do not afford a stable emulsion of DSCG droplets. Both the occurrence and the stability of this unique emulsion system can be rationalized on the basis of the functional groups of the polymer. The different configurations of the liquid crystal (DSCG) droplets were also found to correlate with the strength of the hydrogen bonding that can be formed by the functional groups on the polymer.

Objective To develop a mouse model for the study of the pathophysiologic mechanism and treatment of human bone marrow (BM) failure. Materials and methods Unmanipulated B6D2F1 or CByB6F1 hybrid mice were infused with 10–40 × 10 6 lymph node (LN) cells from their C57BL/6 (B6) parent. Pancytopenia was monitored by cell counting, while marrow damage was assessed by histological staining. Destruction of BM hematopoietic progenitor/stem cells was measured by colony formation in vitro and irradiation protection in vivo. Serum interferon-γ (IFN-γ) concentration was measured by enzyme-linked immunosorbent assay. BM T cell Vβ and Fas expressions were analyzed by flow cytometry. Treatment effects of immunosupressive agents cyclosporine, antithymocyte globulin (ATG), anti-IFN-γ, and anti-tumor necrosis factor-α (anti-TNF-α) were tested. Results Infusion of 30–40 × 10 6 B6 LN cells led to rapid development of severe pancytopenia, BM hypoplasia, and death. Affected mice had drastically reduced hematopoietic progenitor and stem cells. BM of affected mice showed lymphocyte infiltration, oligoclonal T cell expansion, and upregulated Fas expression. Serum IFN-γ concentration increased two- to three-fold. Timed administration of cyclosporine or ATG abrogated pancytopenia. Treatment with anti-IFN-γ antibody reliably rescued mice, and treatment with anti-TNF-α antibody extended animal survival significantly. Conclusion This mouse model indicates that activated lymphocytes and type I cytokines play important roles in marrow destruction in lymphocyte infusion-induced BM failure.

Although living “happily ever after” may still be the conclusion of many children’s stories, the reality of marriage1 is often not so happy or permanent. Statistics show that approximately 50% of marriages in the United States end in divorce and one survey shows that many of the marriages that do not end in divorce appear to be distressed (Baucom, Epstein, Rankin, & Burnett, 1996). Thankfully, couple therapy has been shown to be effective in helping troubled couples and there is a large body of research regarding the functioning of couples and the treatment of couple difficulties (well summarized in Baucom, Shoham, Mueser, Daiuto, & Stickle, 1998; Epstein, 2001; and Epstein & Baucom, 2002).