Increase In Relative Proportion Research Articles

Genetic ancestry and diagnostic yield of exome sequencing in a diverse population

It has been suggested that diagnostic yield (DY) from Exome Sequencing (ES) may be lower among patients with non-European ancestries than those with European ancestry. We examined the association of DY with estimated continental/subcontinental genetic ancestry in a racially/ethnically diverse pediatric and prenatal clinical cohort. Cases (N = 845) with suspected genetic disorders underwent ES for diagnosis. Continental/subcontinental genetic ancestry proportions were estimated from the ES data. We compared the distribution of genetic ancestries in positive, negative, and inconclusive cases by Kolmogorov–Smirnov tests and linear associations of ancestry with DY by Cochran-Armitage trend tests. We observed no reduction in overall DY associated with any genetic ancestry (African, Native American, East Asian, European, Middle Eastern, South Asian). However, we observed a relative increase in proportion of autosomal recessive homozygous inheritance versus other inheritance patterns associated with Middle Eastern and South Asian ancestry, due to consanguinity. In this empirical study of ES for undiagnosed pediatric and prenatal genetic conditions, genetic ancestry was not associated with the likelihood of a positive diagnosis, supporting the equitable use of ES in diagnosis of previously undiagnosed but potentially Mendelian disorders across all ancestral populations.

Effects of aqueous fluoxetine exposure on gut microbiome of adult Pimephales promelas

The microbiome of the gut is vital for homeostasis of hosts with its ability to detoxify and activate toxicants, as well as signal to the immune and nervous systems. However, in the field of environmental toxicology, the gut microbiome has only recently been identified as a measurable indicator for exposure to environmental pollutants. Antidepressants found in effluents of wastewater treatment plants and surface waters have been shown to exhibit antibacterial-like properties in vitro, where some bacteria are known to express homologous proteins that bind antidepressants in vertebrates. Therefore, it has been hypothesized that exposure to antidepressant drugs might affect gut microbiota of aquatic organisms. In this study, the common antidepressant, fluoxetine, was investigated to determine whether it can modulate the gut microbiome of adult fathead minnows. A 28-day, sub-chronic, static renewal exposure was performed with nominal fluoxetine concentrations of 0.01, 10 or 100 μg/L. Using 16S rRNA amplicon sequencing, shifts among the gut-associated microbiota were observed in individuals exposed to the greatest concentration, with greater effects observed in females. These changes were associated with a decrease in relative proportions of commensal bacteria, which can be important for health of fish including bacteria essential for fatty acid oxidation, and an increase in relative proportions of pathogenic bacteria associated with inflammation. Results demonstrate, for the first time, how antidepressants found in some aquatic environments can influence gut microbiota of fishes.

Chronic ischemic heart failure in humans is associated with changes in expression of antigen processing HLA types and activation of T-cells

Abstract Background Sustained pathological inflammation is a hallmark of chronic ischemic heart failure (HF). The short term response to myocardial ischemia has found a protective phenotype for T regulatory cells, yet the extent and consequence of prolonged T-cell activation in chronic ischemic heart failure is unclear. Methods and results Single cell RNA sequencing of circulating immune cells (n=181,712 cells) from healthy (n=8) and heart failure donors (n=8) revealed a relative increase in the proportion of activated T cells in heart failure patients relative to healthy controls. T cell activation molecules of the HLA-DR series genes on antigen presenting cells were found to be strongly associated with heart failure patients in the sequencing data, with decreases in antigen processing genes (HLA-DM series) in heart failure patients. Validation via flow cytometry in 27 healthy controls and 43 heart failure patients showed relative decreases of greater than 30% for HLA-DM to HLA-DRB molecule ratios in HF patients (p=0.0082), suggesting an aberrant antigen presentation to T cells linked to autoimmunity in heart failure patients. Additionally, costimulatory molecules such as ICAM-1 (p=0.004) and activation markers like TREM-1 (p=0.04) were elevated in circulating monocytes cells, which may potentiate T cell activation. Correspondingly, a significant and striking decrease in CD4+ and CD8+ naïve T cells along with increases in combined effector memory and TEMRA cells could be evidenced in heart failure patients. Levels of the T-cell homing marker CCR5 were elevated in patients with heart failure. To gain additional insights into potential functional consequences in the heart, supernatants from 3h LPS stimulated immune cells (peripheral blood mononuclear cells or T cells) were applied to various vascular cell types (endothelial cells, pericytes) and with cardiac organoids for 48h. Apoptosis was increased more than 3 fold following incubation with media derived from peripheral blood cells of HF patients, suggesting that immune cells from heart failure patients can directly and indirectly impair cardiovascular cells. Conclusion Heart failure is marked by sustained increases in relative proportions of effector T cell and TEMRA populations and homing markers suggesting these cells may undergo extravasation to tissues and deteriorate inflamed myocardium - hastening heart failure progression, worsening prognosis. Further research is required to assess how these cells may migrate to myocardial tissues and what interventions could be useful for blocking myocardial damage. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): DZHK, DGK, SFB

Impact of New Clinical Policies during the COVID-19 Pandemic on Clinical Incidents and Complaints at a UK Teaching Hospital.

Background: To investigate any associations between new clinical policies implemented because of the COVID-19 pandemic and harm to patients. Methods: Retrospective data collection of incidents and complaints reported through Datix®, and the Patient Advice and Liaison Service (PALS), respectively. The setting was the Family Health division in a University teaching hospital in the UK. Primary and secondary outcome measures included: the proportion of incidents reported on Datix® from 23 March 2020 to 29 May 2020, compared to the period from 23 March 2019 to 29 May 2019. COVID-19 related incidents and complaints and association with newly published guidelines or pathways from 23 March 2020 to 29 May 2020 were investigated. Results: There was no significant difference in the proportion of overall patient activity resulting in incidents reported on Datix in 2020 (2.08%) compared to 2019 (2.09%), with 98% resulting in no/low harm in 2020. Three incident categories had increases in relative proportions of incidents including the terms “COVID” or “Corona” compared to incidents that did not: “Child death”, “delay/failure to treatment and procedure” and “information governance”. One of the child deaths was a miscarriage and we were unable to link the second child death to a change in clinical policy at this stage. We were only able to link two COVID-19 associated incidents with a pathway or procedural change (one to the Children’s Emergency Department admission pathway and the second to the introduction of virtual antenatal clinics). Eighteen complaints related to COVID-19 were logged. However, at this stage, we are unable to link any of these to a published change in clinical policy. Conclusions: New policies introduced in the division, during the COVID-19 pandemic were associated with similar rates of clinical incidents, when compared with the previous year. There were only two COVID-19-related incidents clearly related to a change in pathways and procedures. Continued surveillance and improved metrics for monitoring the impact of changes to pathways and procedures should be sought with the sustained presence of COVID-19 in clinical areas.

Characterization of reversible and irreversible foulants in membrane bioreactor (MBR) for eucalyptus pulp and paper mill wastewater treatment using fluorescence regional integration

Abstract Membrane bioreactors (MBRs) are an alternative technology to achieve high quality effluent for water reuse for eucalyptus pulp and paper (P&P) industries, but membrane fouling is still a challenging factor that restricts its application. This study aims to investigate characteristics of reversible and irreversible fouling of the submerged polyvinylidene fluoride (PVDF) MBR for treating eucalyptus P&P wastewater. Six batches of MBR experiments with different mixed liquor suspended solids (MLSS) concentrations were performed. The membrane cleaning protocol included three-steps: backwashing with DI water and subsequently NaOH solution, and then soaking with NaOCl solution, for which achieved 54.6 ± 11 %, 76.6 ± 10 % and 97.8 ± 2 % of flux recovery, respectively. Formation of a cake layer comprised of organic substance and soluble microbial products (SMPs), mostly polysaccharides and proteins, was the main contributor to flux decline of the MBR. Concentration of proteins in the irreversible foulant was found to be positively correlated with degree of membrane fouling. Using fluorescence excitation-emission matrix (EEM) with fluorescence regional integration (FRI) analysis revealed that, during MBR treatment, protein-like substances in the permeate water significantly decreased, simultaneously with the increase in relative proportions of humic-like and fulvic-like substances. Further analysis on the irreversible foulants showed that protein-like substances, including tyrosine-like and tryptophan-like substances, were disproportionately more important to membrane fouling than humic-like substances relative to their respective concentration in raw P&P wastewater.

An Integrated Infant and Young Child Feeding and Small-Quantity Lipid-based Nutrient Supplementation Program Is Associated with Improved Gross Motor and Communication Scores of Children 6-18 Months in the Democratic Republic of Congo

To evaluate the impact of an infant, young child feeding practices-small-quantity lipid nutrient supplements (SQ-LNS) intervention on child development scores in children aged 6-18months in the Katanga Province, Democratic Republic of Congo (DRC). We analyzed data of 2595 children from 2 health zones in a quasi-experimental design with preimplementation and postimplementation surveys to evaluate program impact on child development scores. Standard care was received in the comparison health zone and the intervention health zone received standard care plus enhanced infant, young child feeding practices with a monthly supply of 28 SQ-LNS sachets for up to 1year. Program exposure and communication and motor domains of the Ages and Stages questionnaire were collected to assess changes in child development scores. A quasi-intent-to-treat and adjusted difference-in-difference analyses were used to quantify impact of the enhanced compared with the standard package. In adjusted models contrasting endline with baseline, there was a greater relative increase in proportion of children with normal communication (difference-in-difference, +13.7% [95%CI, 7.9-19.6; P<.001] and gross motor scores, +7.4% [95% CI: 1.3-13.5; P<.001]) in the intervention vs comparison health zones. Further, in separate analyses among children of intervention health zone at endline, each additional SQ-LNS distribution was associated with +0.09 (95% CI, 0.03-0.16) z-score unit increase in gross motor scores (P<.01). The integrated infant, young child feeding practice-SQ-LNS intervention was positively associated with larger relative improvements in measures of child communication and motor development in the Katanga province of DRC.

Hydrometeorological processes in semi-arid western India: insights from long term isotope record of daily precipitation

Long term (2005–2016) daily precipitation isotope data (δ18O, δD and d-excess) from Ahmedabad in semi-arid Western India are examined in light of various meteorological parameters and air parcel trajectories to identify prominent patterns in the isotopic character and discern the underlying hydrometeorological processes. One of the most prominent and systematic annual patterns is the isotopic depletion (average δ18O: − 2.5‰ in Jun–Jul; − 5.2‰ in Aug–Sept) in the second half of the Indian Summer Monsoon (ISM), which is observed in the 11 out of the 12 years of this study. Four geographically feasible causal factors have been examined if they contribute to observed late monsoon isotopic depletion. These factors are: (1) increased contribution of terrestrially recycled vapor; (2) intra-seasonal change in sea-surface, surface-air and cloud base temperatures; (3) increased rain-out fraction from marine vapor parcel; and (4) increase in relative proportion of convective rain. It is inferred from the present study that isotopic depletion in the second half of ISM is associated with: (1) increased contribution (45% from 36%) of terrestrially recycled moisture; (2) 1.9° C lower cloud base temperature; (3) increased rainout fraction due to decreased wind velocity (6.9 m/s from 8.8 m/s); and (4) an increase of 22.3% in the proportion of convective rain. Daily rain events with atypical isotopic composition (20‰ < d-excess < 0‰) are ascribed mainly to local weather perturbations causing sudden updraft of moist air facilitating terrestrial recycling of water vapor.

Characterization of bacterial communities in wastewater with enhanced taxonomic resolution by full-length 16S rRNA sequencing

Wastewater treatment is crucial to environmental hygiene in urban environments. However, wastewater treatment plants (WWTPs) collect chemicals, organic matter, and microorganisms including pathogens and multi-resistant bacteria from various sources which may be potentially released into the environment via WWTP effluent. To better understand microbial dynamics in WWTPs, we characterized and compared the bacterial community of the inflow and effluent of a WWTP in Berlin, Germany using full-length 16S rRNA gene sequences, which allowed for species level determination in many cases and generally resolved bacterial taxa. Significantly distinct bacterial communities were identified in the wastewater inflow and effluent samples. Dominant operational taxonomic units (OTUs) varied both temporally and spatially. Disease associated bacterial groups were efficiently reduced in their relative abundance from the effluent by the WWTP treatment process, except for Legionella and Leptospira species which demonstrated an increase in relative proportion from inflow to effluent. This indicates that WWTPs, while effective against enteric bacteria, may enrich and release other potentially pathogenic bacteria into the environment. The taxonomic resolution of full-length 16S rRNA genes allows for improved characterization of potential pathogenic taxa and other harmful bacteria which is required to reliably assess health risk.

The Fate of Platinum-Group Minerals in the Exogenic Environment—From Sulfide Ores via Oxidized Ores into Placers: Case Studies Bushveld Complex, South Africa, and Great Dyke, Zimbabwe

Diverse studies were performed in order to investigate the behavior of the platinum-group minerals (PGM) in the weathering cycle in the Bushveld Complex of South Africa and the Great Dyke of Zimbabwe. Samples were obtained underground, from core, in surface outcrops, and from alluvial sediments in rivers draining the intrusions. The investigations applied conventional mineralogical methods (reflected light microscopy) complemented by modern techniques (scanning electron microscopy (SEM), mineral liberation analysis (MLA), electron-probe microanalysis (EPMA), and LA-ICPMS analysis). This review aims at combining the findings to a coherent model also with respect to the debate regarding allogenic versus authigenic origin of placer PGM. In the pristine sulfide ores, the PGE are present as discrete PGM, dominantly PGE-bismuthotellurides, -sulfides, -arsenides, -sulfarsenides, and -alloys, and substantial though variable proportions of Pd and Rh are hosted in pentlandite. Pt–Fe alloys, sperrylite, and most PGE-sulfides survive the weathering of the ores, whereas the base metal sulfides and the (Pt,Pd)-bismuthotellurides are destroyed, and ill-defined (Pt,Pd)-oxides or -hydroxides develop. In addition, elevated contents of Pt and Pd are located in Fe/Mn/Co-oxides/hydroxides and smectites. In the placers, the PGE-sulfides experience further modification, whereas sperrylite largely remains a stable phase, and grains of Pt–Fe alloys and native Pt increase in relative proportion. In the Bushveld/Great Dyke case, the main impact of weathering on the PGM assemblages is destruction of the unstable PGM and PGE-carriers of the pristine ores and of the intermediate products of the oxidized ores. Dissolution and redistribution of PGE is taking place, however, the newly-formed products are thin films, nano-sized particles, small crystallites, or rarely µm-sized grains primarily on substrates of precursor detrital/allogenic PGM grains, and they are of subordinate significance. In the Bushveld/Great Dyke scenario, and in all probability universally, authigenic growth and formation of discrete, larger PGM crystals or nuggets in the supergene environment plays no substantial role, and any proof of PGM “neoformation” in a grand style is missing. The final PGM suite which survived the weathering process en route from sulfide ores via oxidized ores into placers results from the continuous elimination of unstable PGM and the dispersion of soluble PGE. Therefore, the alluvial PGM assemblage represents a PGM rest spectrum of residual, detrital grains.

STAT3 antisense oligonucleotide AZD9150 in a subset of patients with heavily pretreated lymphoma: results of a phase 1b trial

BackgroundThe Janus kinase (JAK) and signal transduction and activation of transcription (STAT) signaling pathway is an attractive target in multiple cancers. Activation of the JAK-STAT pathway is important in both tumorigenesis and activation of immune responses. In diffuse large B-cell lymphoma (DLBCL), the transcription factor STAT3 has been associated with aggressive disease phenotype and worse overall survival. While multiple therapies inhibit upstream signaling, there has been limited success in selectively targeting STAT3 in patients. Antisense oligonucleotides (ASOs) represent a compelling therapeutic approach to target difficult to drug proteins such as STAT3 through of mRNA targeting. We report the evaluation of a next generation STAT3 ASO (AZD9150) in a non-Hodgkin’s lymphoma population, primarily consisting of patients with DLBCL.MethodsPatients with relapsed or treatment refractory lymphoma were enrolled in this expansion cohort. AZD9150 was administered at 2 mg/kg and the 3 mg/kg (MTD determined by escalation cohort) dose levels with initial loading doses in the first week on days 1, 3, and 5 followed by weekly dosing. Patients were eligible to remain on therapy until unacceptable toxicity or progression. Blood was collected pre- and post-treatment for analysis of peripheral immune cells.ResultsThirty patients were enrolled, 10 at 2 mg/kg and 20 at 3 mg/kg dose levels. Twenty-seven patients had DLBCL. AZD9150 was safe and well tolerated at both doses. Common drug-related adverse events included transaminitis, fatigue, and thrombocytopenia. The 3 mg/kg dose level is the recommended phase 2 dose. All responses were seen among DLBCL patients, including 2 complete responses with median duration of response 10.7 months and 2 partial responses. Peripheral blood cell analysis of three patients without a clinical response to therapy revealed a relative increase in proportion of macrophages, CD4+, and CD8+ T cells; this trend did not reach statistical significance.ConclusionsAZD9150 was well tolerated and demonstrated efficacy in a subset of heavily pretreated patients with DLBCL. Studies in combination with checkpoint immunotherapies are ongoing.Trial registrationRegistered at ClinicalTrials.gov: NCT01563302. First submitted 2/13/2012.

Tidal Stage Changes in Structure and Diversity of Intertidal Benthic Diatom Assemblages: a Case Study from Two Contrasting Charleston Harbor Flats

Benthic microalgae are key contributors to near-shore food webs and sediment stabilization. Temporal variability in microalgal biomass and production throughout the tidal cycle has been well documented; however, due to limitations of traditional methods of analysis patterns of community composition and diversity over such time scales have not been revealed. To explore the latter and better understand how short-term changes throughout the tidal cycle may affect community functioning, we compared benthic diatom composition and diversity over tidal stage shifts. We employed two disparate molecular techniques (denaturing gel gradient electrophoresis with Sanger DNA sequencing of excised bands and high-throughput community metagenome sequencing) to characterize diatom assemblages in representative muddy and sandy intertidal sites in Charleston Harbor, SC, USA. In support of prior studies, we found higher diatom diversity in sandbar as compared to mudflat sediments. Spatial differences were stronger relative to tidal temporal differences, although diversity metrics generally were highest after prolonged tidal immersion as compared to low-tide emersion or just after immersion at flood tide. Composition of the diatom assemblage differed markedly between sites, with species in genera Halamphora, Amphora, and Navicula dominating the sandbar, whereas Cyclotella, Skeletonema, and Thalassiosira were the most prevalent genera on the mudflat. Diatom composition differed by tidal stage, with assemblages during low-tide exposure distinct from samples taken after immersion. Both sandbar and mudflat sediments exhibited increases in relative proportion of epipelic diatoms and decreases in planktonic taxa during low-tide exposure. Our findings of short-term changes in species composition and dominance could inform primary productivity models to better estimate understudied diatom contributions in heterogeneous and highly variable tidal systems.

Pericentromeric Regions Are Refractory To Prompt Repair after Replication Stress-Induced Breakage in HPV16 E6E7-Expressing Epithelial Cells

Chromosomal instability is the major form of genomic instability in cancer cells. Amongst various forms of chromosomal instability, pericentromeric or centromeric instability remains particularly poorly understood. In the present study, we found that pericentromeric instability, evidenced by dynamic formation of pericentromeric or centromeric rearrangements, breaks, deletions or iso-chromosomes, was a general phenomenon in human cells immortalized by expression of human papillomavirus type 16 E6 and E7 (HPV16 E6E7). In particular, for the first time, we surprisingly found a dramatic increase in the proportion of pericentromeric chromosomal aberrations relative to total aberrations in HPV16 E6E7-expressing cells 72 h after release from aphidicolin (APH)-induced replication stress, with pericentromeric chromosomal aberrations becoming the predominant type of structural aberrations (∼70% of total aberrations). In contrast, pericentromeric aberrations accounted for only about 20% of total aberrations in cells at the end of APH treatment. This increase in relative proportion of pericentromeric aberrations after release from APH treatment revealed that pericentromeric breaks induced by replication stress are refractory to prompt repair in HPV16 E6E7-expressing epithelial cells. Telomerase-immortalized epithelial cells without HPV16 E6E7 expression did not exhibit such preferential pericentromeric instability after release from APH treatment. Cancer development is often associated with replication stress. Since HPV16 E6 and E7 inactivate p53 and Rb, and p53 and Rb pathway defects are common in cancer, our finding that pericentromeric regions are refractory to prompt repair after replication stress-induced breakage in HPV16 E6E7-expressing cells may shed light on mechanism of general pericentromeric instability in cancer.

M2 macrophages exhibit higher sensitivity to oxLDL-induced lipotoxicity than other monocyte/macrophage subtypes

BackgroundIn obesity, phenotypic switches occur in macrophage populations such that the predominantly M2-polarised anti-inflammatory state seen in lean individuals changes to a predominantly M1-polarised pro-inflammatory state in those who are obese. However, the mechanisms by which these phenotypic shifts occur have not yet been fully elucidated.ResultsThe effects of oxLDL (1-40 μg/ml; 24 h) on several parameters relevant to the Unfolded Protein Response (UPR)-mediated lipotoxic effects of oxLDL (disruption of ER Ca2+ handling; activation of the UPR transcription factor XBP-1; upregulation of the UPR target genes BiP and CHOP; apoptosis; cell viability) were investigated in human primary monocyte-derived macrophages, and also in monocyte-macrophages derived from the THP-1 monocytic cell line. A consistent pattern was observed: M2-polarised macrophages were more sensitive to the lipotoxic effects of oxLDL than either non-polarised macrophages or non-differentiated monocytic cells. Specifically, M2-polarised macrophages were the only cell type to undergo significantly increased apoptosis (Primary cells: 1.23 ± 0.01 basal; THP-1-derived: 1.97 ± 0.12 basal; P < 0.05 in both cases) and decreased cell viability (Primary cells: 0.79 ± 0.04 basal; THP-1-derived: 0.67 ± 0.02 basal; P < 0.05 in both cases) when exposed to oxLDL levels similar to those seen in overweight individuals (ie. 1 μg/ml).ConclusionsWe propose that the enhanced susceptibility of M2-polarised macrophages to lipotoxicity seen in the present in vitro study could, over time, contribute to the phenotypic shift seen in obese individuals in vivo. This is because a higher degree of oxLDL-induced lipotoxic cell death within M2 macrophages could contribute to a decrease in numbers of M2 cells, and thus a relative increase in proportion of non-M2 cells, within macrophage populations. Given the pro-inflammatory characteristics of a predominantly M1-polarised state, the data presented here may constitute a useful contribution to our understanding of the origin of the pro-inflammatory nature of obesity, and of the pathogenesis of obesity-associated inflammatory disorders such as Type 2 Diabetes and atherosclerosis.

An environmentally relevant endocrine-disrupting antiandrogen, vinclozolin, affects calling behavior of male Xenopus laevis

Vinclozolin (VIN) is an antiandrogenic model substance as well as a common fungicide that can affect the endocrine system of vertebrates. The objective of this study was to investigate how VIN affects mate calling behavior of South African clawed frogs ( Xenopus laevis) and whether it is effective at environmentally relevant concentrations. Male X. laevis were injected with human chorionic gonadotropin (hCG) to stimulate their androgen-controlled mate calling behavior and were treated with VIN at concentrations of 10 − 6, 10 − 8 and 10 − 10 M. VIN at 10 − 6 M reduced calling activity. Furthermore, the vocalization composition of VIN-treated X. laevis was altered. The call types advertisement calls and chirping are uttered by reproductively active males, whereas the call types growling, ticking, and rasping indicate a sexually unaroused state of a male. VIN at any of the tested concentrations led to a decrease in utterance of calls, which indicate a sexually aroused state of the males, and an increase in relative proportions of calls, indicating a sexually unaroused state of the males. Additionally, the mean duration of clicks and the number of accentuated clicks during the advertisement calls decreased at all concentrations of VIN. No significant differences were observed in any other temporal or spectral calling parameters between the treatments. This study illustrates that exposure to the antiandrogen VIN might result in a reduced reproductive success by altering mate calling behavior of X. laevis. Moreover, it suggests that the behavioral parameters examined in this study can be used as sensitive biomarkers for detecting antiandrogenic endocrine disrupting compounds in amphibians.

Solid-state NMR characterization of switchgrass cellulose after dilute acid pretreatment

Background: Switchgrass (Panicum virgatum L.) is being considered as a potential feedstock for bioethanol production and extensive research is ongoing to establish the optimum pretreatment conditions for switchgrass. Cellulose comprises the most abundant biopolymer in the biosphere and its crystalline/ultrastructure is considered to be related to biomass recalcitrance. This study investigates the effects of dilute acid pretreatment on ultrastructural features of switchgrass cellulose. Results: In this study, switchgrass was pretreated in a pilot-scale reactor at 190°C (0.05 g sulfuric acid per gram of dry switchgrass) with 25% total solid loading and a reactor residence of 1 min. Cellulose was isolated from the pretreated and untreated switchgrass. The impact of pretreatment on the ultrastructure of cellulose was determined by solid-state cross polarization/magic angle spinning 13C NMR spectroscopy. Switchgrass demonstrated a preferable degradation of amorphous cellulose regions during dilute acid pretreatment. The pretreated switchgrass cellulose showed an 18% increase in crystallinity index when compared with the untreated switchgrass. Line-fitting analysis of the C-4 region of 13C NMR spectra revealed that the relative proportion of crystalline and paracrystalline celluloses in switchgrass was observed to increase after dilute acid pretreatment, accompanied with a concurrent decrease of the relative abundance of fibril surface cellulose. Conclusion: After dilute acid pretreatment, most of the hemicellulose in switchgrass was removed. The amorphous cellulose regions in switchgrass were degraded preferably during dilute acid pretreatment and the cellulose crystallinity index of pretreated switchgrass increased. Pretreated switchgrass had an increase in relative proportion of crystalline and paracrystalline cellulose in comparison to the starting material.

Seasonal acquisition of chill tolerance and restructuring of membrane glycerophospholipids in an overwintering insect: triggering by low temperature, desiccation and diapause progression

Adults of the insect Pyrrhocoris apterus acquire chill tolerance through the process of autumnal acclimatization. Field and laboratory experiments were conducted to separate the triggering effects of low temperatures, desiccation and diapause progression on the physiological characteristics related to chill tolerance with emphasis on the restructuring of glycerophospholipid (GPL) composition. Changes in relative proportions of major molecular species of glycerophosphoethanolamines (GPEtns) and glycerophosphocholines (GPChols) in thoracic muscle and fat body tissues were followed using HPLC coupled to electrospray ionisation mass spectrometry. The increase in relative proportion of 1-palmitoyl-2-linoleyl-sn-GPEtn at the expense of 1,2-dilinoleyl-sn-GPChol was the most prominent feature of the complex change observed in both tissues during autumnal acclimatization in the field. The relative proportion of total GPEtns increased, while the proportion of total GPChols decreased. The relative proportion of unsaturated fatty acyls slightly decreased. A similar restructuring response was seen during acclimatization in the field and cold acclimation in the laboratory. By contrast, the GPL changes related to desiccation and diapause progression were relatively small, differed qualitatively from the cold-acclimation response, and were accompanied with no increase of chill tolerance. Other features of autumnal acclimatization, i.e. depression of supercooling capacity and accumulation of polyhydric alcohols, were also triggered solely by low temperatures.

Increased ratio of saturated to unsaturated C18 fatty acids in colonic adenocarcinoma: Implications for cryotherapy and lipid raft function

Mammalian fatty acid synthase (FASE) overexpression has been shown in a number of human malignancies including colonic adenocarcinoma. Since FASE synthesizes only saturated fatty acids, we hypothesized that cancer cells have a greater proportion of long-chain saturated fatty acids. We studied and found an unequivocal increase in saturated C18 fatty acid (stearic acid) in colonic adenocarcinoma compared to adjacent normal colonic mucosa. The increase is even more striking when measured as a ratio of stearic acid to the unsaturated C18 fatty acids (oleic acid and linoleic acid). This change in fatty acid composition of the cancer cells should significantly alter their physical and biological properties. The increase in relative proportion of saturated fatty acids should make the cancer cells more susceptible to cryodamage and measurement of fatty acid composition of cancer cells may help individualize the temperature for cryotherapy. Also, the lipid alterations may affect the structure and functions of lipid rafts, which may enable the cancer cells to affect signaling mechanisms such as those involved in cell growth and apoptosis. Dietary or therapeutic interventions targeting lipid rafts may thus be an option for cancer treatment.

Changes in Thymopoiesis in Myasthenia Gravis

This study was undertaken to investigate T-cell maturation in hyperplastic thymi of patients suffering from myasthenia gravis (MG). For this purpose, the expression of the major differentiational molecules (CD4, CD8, and CD3/TCRαβ) and that of the regulatory and activation molecules on thymocytes from MG patients and control subjects were estimated by flow cytometric analysis. In the MG patients the increase in relative proportion of immature (CD4−8− TCRαβ−) and the most mature (CD4+8− TCRαβhigh and CD4−8− TCRhigh encompassing immunoregulatory NKT) thymocytes followed by a decrease in that of CD4+8+CD3−/TCRαβ− cells was found. Furthermore, in these patients the relative proportion of CD4+HLA-DR+ and CD4+71+ cells was increased, whereas that of CD4+25+ cells was slightly, but significantly, decreased (reflecting, most likely, decreased contribution of T reg cells bearing this phenotype). Moreover, in MG thymi the percentage of CD45RA+ cells was reduced indicating changes in the selection processes. In keeping with this finding the reduced thymocyte apoptotic index and percentage of cells bearing apoptosing (CD4−8− TCRαβlow) phenotype were detected. In conclusion, the study demonstrates substantial changes in intrathymic differentiation of T cells in hyperplastic MG thymi and suggests alterations in selection events providing an increased escape of potentially autoreactive T-cell clones, on one side, and an altered maturation and/or selection of immunoregulatory cells (NKT and CD4+8−25+ T reg cells) keeping these cell clones under control, on the other side.

Activation of protein kinase C increases proteoglycan synthesis in immature rat Sertoli cells

In order to determine the signal transduction pathways involved in the regulation of proteoglycan (PG) synthesis in immature rat Sertoli cells (SC), we have examined the effect of the tumor promoter phorbol ester PMA (phorbol myristate acetate) on [ 35S]sulfate and [ 3H]glucosamine incorporation into PG molecules neosynthesized by cultured rat SC. PMA induced a dose- and time-dependent stimulation of labeled cell-associated PG as determined by quantitative solid phase assay. The overall effect of PMA resulted from enhancement of both glycosylation and catabolism of cell PG, this latter effect leading to a drastic decrease of their residence time in the membrane. Besides these quantitative effects, activation of protein kinase C by PMA induced qualitative changes as reflected by increase in relative proportion of heparan sulfate PG (HSPG) in cell membrane PG. In light of our previous results suggesting an inverse relationship between PG synthesis and FSH responsiveness in immature rat Sertoli cells, the PMA-induced upregulation of cell membrane PG, and particularly HSPG, could constitute one mechanism involved in the repression of FSH-stimulated steroidogenesis induced by PKC activation.

Cinétiques des variations de la composition protéique du lait au cours d'une mammite expérimentale à Escherichia coli

Sequential changes in milk protein composition after experimental Escherichia coli mastitis. Milk protein composition was determined between 0 and 72 h after infection during experimentally induced E. coli mastitis of seven cows. In milk from infected quarters, contents of non protein nitrogen, total proteins and soluble proteins were significantly higher after 14 h and a max- imum was reached at 48 h after infection (+44 %, + 104 % and +574 % respectively). Casein content independently from time was reduced of 20 %. Proportions of 13- and œ-caseins were significantly lower at 24 h (-49 %) and between 48 and 72 h (--D2 %) respectively. Proportion of pH 4.6 insoluble pep- tides including y-caseins reached a maximum between 24 and 72 h (+580 %). These different kinet- ics can be explained by sequential origin ofproteolysis; plasmin activity could be maximal before max- imal activity of somatie cell proteases. Protein composition in milk from control quarters was also greatly modified. Regardless of time, soluble protein content increased (+68 %) and casein content decreased (-21 %). Significative proteolysis of l3-casein occurred at 72 h (-25 %) accompanied by increase in relative proportion of pH 4.6 insoluble peptides (+50 %); this was probably due to higher milk plasmin activity levels. Therefore, interdependance was notieed between infected and unin- fected quarters, probably through systemic response to E. coli infection. © InralElsevier, Paris.