Corticosteroid dose and the risk of opportunistic infection in a national systemic lupus erythematosus cohort.
Objective This study investigated whether the incidence of opportunistic infection differed in systemic lupus erythematosus patients who received different doses of corticosteroids. Methods We included patients with diagnosed systemic lupus erythematosus from 1997 to 2010 using Taiwan national health insurance data. The index day for systemic lupus erythematosus patients was 3 months after the systemic lupus erythematosus diagnosis. A non-steroid cohort was matched 4:1 with the steroid cohort according to age, sex and index day. The end of the follow-up period was the day of opportunistic infection diagnosis, 1 year after the index day, or death. Results The overall cumulative incidence of opportunistic infection was 136-fold higher in the steroid cohort than in the non-steroid cohort. The adjusted hazard ratio for developing mycobacterium infection in the steroid cohort was 11, and the adjusted hazard ratio for developing herpes zoster was 43.6 compared to the non-steroid cohort after adjusting for immunosuppressive agents and comorbidities. The adjusted hazard ratio value for opportunistic infection was 1.40 (95% confidence interval (CI) 0.78-2.51) for a daily prednisone-equivalent dose of 7.5-15 mg, 1.72 (95% CI 1.02-2.91) for 15-30 mg, 1.96 (95% CI 1.17-3.28) for 30-60 mg and 2.24 (95% CI 1.26-4.00) for over 60 mg compared with low-dose steroids (<7.5 mg). Conclusion This study confirmed that the risk of opportunistic infection is higher in systemic lupus erythematosus patients treated with steroids in the first 3 months after diagnosis versus those not treated with steroids. Medium and high doses were associated with a higher risk of opportunistic infection compared with low doses. However, there was no controlling for disease activity, making it hard to know if increases in infection were due to disease itself or corticosteroids.


The long-term effect of a zinc acetate and chlorhexidine diacetate containing mouth rinse on intra-oral halitosis-A randomized clinical trial.
To evaluate the long-term effects of a zinc acetate and chlorhexidine diacetate mouth rinse (Zn/CHX) on intra-oral halitosis. Forty-six adults with intra-oral halitosis were randomized into a 6-month, double-blind, placebo-controlled clinical study. The presence of intra-oral halitosis was evaluated at baseline, 3 and 6 months after treatment by assessment of organoleptic score (OLS) and by total volatile sulphur compounds (T-VSC), hydrogen sulphide (H2 S) and methyl mercaptan (MM) concentrations in exhaled air. A Zn/CHX mouth rinse provided significantly better control of intra-oral halitosis than a placebo mouth rinse. At 3 and 6 months, individuals rinsing with the Zn/CHX rinse presented with reductions of the OLS, T-VSC (p < .01, respectively), H2 S (p < .001), and MM (p < .01) in subjects' exhaled air. At 6 months, 68.2% of individuals using the Zn/CHX rinse experienced a 1 or 2 category improvement in OLS compared with 19.1% of placebo-treated subjects. 91% of subjects in the Zn/CHX group were categorized as being effectively treated for intra-oral halitosis (i.e. H2 S < 112 ppb), compared to 43% in the placebo group. Zn/CHX mouth rinse provides effective long-term efficacy against intra-oral halitosis, assessed both objectively and subjectively. With regular rinsing, the effect was sustained for 6 months.


HLA-A2, HLA-B44 and HLA-DR15 are associated with lower risk of BK viremia.
Human leucocyte antigens (HLAs) modulate immunity to polyomavirus BK (BKV). Identification of HLAs that alter the course of infection will facilitate risk stratification, and customization of pre-emptive intervention strategies. We performed a retrospective cohort study with 998 kidney transplant patients with BKV infection status confirmed by polymerase chain reaction (PCR). Clinical parameters and donor-recipient matching for specific HLAs were examined in relation to occurrence of viremia. An emphasis was placed on donor-recipient matching rather than the actual frequency of specific HLA-alleles, since a successful immune response requires sharing of HLAs between a virus-infected target cell and the anti-viral effector cell. Using multivariate statistics, low risk of BK viremia was associated with matching of HLA-A2 [hazard ratio (HR) 0.51, 95% confidence interval (CI) 0.28-0.85], HLA-B44 (HR 0.31, 95% CI 0.076-0.85) and HLA-DR15 (HR 0.35, 95% CI 0.084-0.93) (P < 0.05), whereas high risk of viremia was associated with male gender (HR 2.38, 95% CI 1.46-4.09, P < 0.001). HLAs that associated with a lower predisposition to the development of BK viremia have been identified. Evaluation of donor-recipient mismatching for these HLAs could potentially be used to (i) fine tune virus screening strategies for BKV in individual patients and (ii) facilitate discovery of major histocompatibility complex (MHC) class I and II binding peptides that can elicit clinically meaningful BKV-specific immunity.


Short-term stimulation of Na+-dependent amino acid transport by dibutyryl cyclic AMP in hepatocytes. Characteristics and partial mechanism.
The short-term protein-synthesis-independent stimulation of alanine transport in hepatocytes was further investigated. Cyclic AMP increased the Vmax. of alanine transport. Amino acid transport via systems A, ASC and N was stimulated. A good correlation was found between the initial rate of transport and the cell membrane potential as calculated from the distribution of Cl-. Cyclic AMP increased the rate of alanine transport, stimulated Na+/K+ ATPase (Na+/K+-transporting ATPase) activity and caused membrane hyperpolarization. The time courses and cyclic AMP dose-dependencies of all three effects were similar. Ouabain abolished the effect of cyclic AMP on Cl- distribution and on transport of alanine. The effect of cyclic AMP on alanine transport and Cl- distribution was mimicked by the antibiotic nigericin; the effect of nigericin was also abolished by ouabain. It is concluded that the effect of cyclic AMP on transport is mediated via membrane hyperpolarization. It is suggested that the primary action of cyclic AMP is to increase the activity of an electroneutral Na+/K+-exchange system in the liver cell plasma membrane, thus hyperpolarizing the membrane by stimulating the electrogenic Na+/K+ ATPase.


[Differentiated thyroid cancer on the thyroglossal duct].
Over one hundred cases of carcinomas located on the thyroglossal duct have been reported worldwide. Most of these tumors originally were differentiated thyroid cancers. Their biological behavior is the same as that of tumors of the thyroid gland itself. On the basis of one case discovered in an 11-year-old girl, who presented with regional and systemic metastases, we discuss the various aspects of their origin, diagnosis and treatment. As far as the latter is concerned, we think that these tumors can not always be treated by exeresis of the thyroglossal duct. On the basis of the analysis of situations in which these tumors may be encountered, we propose a carcinological surgery, which might possibly be more aggressive, on the thyroid and cervical lymphatic tissues.


Clinical and Microbiological Characteristics of Proven Invasive Aspergillosis Due to Rare/Cryptic Species in Allogeneic Hematopoietic Stem Cell Transplant Recipients.
There are few reports on the clinical course of proven invasive aspergillosis (IA) due to rare/cryptic species in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients. We retrospectively reviewed the electronic medical records of patients who underwent allo-HSCT between January 2012 and December 2018. Of 934 allo-HSCT recipients, 10 were diagnosed with proven IA and 61 were diagnosed with probable IA. DNA sequencing was performed in cases of proven IA, and Aspergillus could be identified to the species level in 8 of the 10 cases. Three were due to A. fumigatus, and 5 were due to rare/cryptic Aspergillus species, namely, A. turcosus, A. felis, A. viridinutans, A. nidulans, and A. calidoustus. In these 8 patients, no patients with IA due to A. fumigatus died, whereas 3 of the 5 with IA due to rare/cryptic species died within 12 weeks. The 2 surviving cases of IA due to rare/cryptic species were treated with surgical resection and antifungal treatment. Susceptibility testing for cryptic species in 4 cases showed an amphotericin B MIC > 1 mg/L in 3 cases, itraconazole MIC > 1 mg/L in 2 cases, and voriconazole MIC > 1 mg/L in 2 cases. In conclusion, more than half of the causative pathogens of proven IA were rare/cryptic species, so it is important to accurately identify the Aspergillus species. In addition, surgical treatment might be an important option in cases of proven IA, given the possibility that the causative organisms are azole-resistant A. fumigatus or rare/cryptic species.


Learning the lessons from banding appeals: evidence based guidance for running junior doctor rotas.
Designing and running robust junior doctor rotas is a challenging task and much previous advice has been based on consensus or anecdote. This paper aims to discern the most frequently occurring problems with trainee working patterns and produce evidence-based guidance for implementing and running contract-compliant rotas. A total of 35 secondary care trusts in North West England were invited to supply information on pay banding appeals requested under the New Deal junior doctor contract. Of these, 15 (43 per cent) participated with data from 35 appeals between 2004 and 2012. A thematic analysis was undertaken to discern the commonly occurring causes of contractual breaches. A total of 83 per cent (n = 29/35) of appeals were based on data showing the rota to be non-compliant with the contract (band 3), with the remainder being compliant with the contract but not in keeping with the pay banding currently assigned. Inability to take adequate natural breaks was the most frequently cited cause of rota non-compliance. Where underlying reasons were given for breaches of hours/rest limits they clustered around 20 themes, the top four being poor or absent dialogue between HR and doctors, excessive workload, inappropriately timed ward rounds, and inadequate or non-existent bleep policies. This is the first analysis of banding appeals under the UK junior doctor contract. The findings show that problems with rotas cluster around specific themes. The authors provide recommendations to target these so as to avoid financially detrimental contract breaches and trainee dissatisfaction.


Accuracy of Radiographic Displacement Measurement in a Pelvic Ring Injury Model.
Pelvic ring injury outcome studies rely on radiographic assessment. To date, no study investigates the accuracy of radiographic measurement. The aim of this study was to assess the accuracy and interobserver reliability of pelvic ring displacement measurement in an injury model. We hypothesize that current radiographic measurement methods do not accurately quantify the three-dimensional pelvic ring displacement. Ten orthopaedic traumatologists evaluated 12 pelvic ring injury model displacements using AP, inlet, and outlet radiographs and axial CT images. Observers completed a survey of demographic and treatment approach strategies. Radiographic displacement measurements in axial, coronal, and sagittal planes were analyzed for accuracy using. Absolute displacement measurements were categorized with Matta and Tornetta grading system for Fleiss Kappa inter-reliability correlation evaluation. The mean age of orthopaedic traumatologists was 47.5 years (range 36 to 59) with a mean 15.3 years (range 4 to 27) of pelvic fracture surgery experience. Radiographic measurement of isolated uniplanar of pelvic displacement in axial, sagittal, or coronal plane alone was more accurate than multiplanar pelvic displacements with more than one plane of displacement, 6.6 ± 5.7 mm error compared with 9.6 ± 6.3 mm error, respectively (P = 0.0035). Measurement accuracy was greater with isolated coronal plane (4 ± 3.5 mm error) compared with isolated axial plane (9.9 ± 7.1 mm error) or isolated sagittal plane displacement (6.7 ± 4 mm error). Interrater reliability for the radiographic displacement measurement by observers showed an overall poor agreement with 0.24. Radiographic displacement measurement in these modeled pelvic ring injuries has notable inaccuracy among various measurement methods. Coronal and sagittal plane radiographic displacement measurements are more accurate compared with axial plane measurement. The reporting of radiographic displacement measurement outcomes in clinical research studies should be critically evaluated, and standardization of pelvic ring injury displacement may not be achievable with radiography. Level V.


How methodological changes have influenced our understanding of population structure in threatened species: insights from tiger populations across India.
Unprecedented advances in sequencing technology in the past decade allow a better understanding of genetic variation and its partitioning in natural populations. Such inference is critical to conservation: to understand species biology and identify isolated populations. We review empirical population genetics studies of Endangered Bengal tigers within India, where 60-70% of wild tigers live. We assess how changes in marker type and sampling strategy have impacted inferences by reviewing past studies, and presenting three novel analyses including a single-nucleotide polymorphism (SNP) panel, genome-wide SNP markers, and a whole-mitochondrial genome network. At a broad spatial scale, less than 100 SNPs revealed the same patterns of population clustering as whole genomes (with the exception of one additional population sampled only in the SNP panel). Mitochondrial DNA indicates a strong structure between the northeast and other regions. Two studies with more populations sampled revealed further substructure within Central India. Overall, the comparison of studies with varied marker types and sample sets allows more rigorous inference of population structure. Yet sampling of some populations is limited across all studies, and these should be the focus of future sampling efforts. We discuss challenges in our understanding of population structure, and how to further address relevant questions in conservation genetics. This article is part of the theme issue 'Celebrating 50 years since Lewontin's apportionment of human diversity'.


Cognitive performance and quality of life in bipolar disorder.
In patients with bipolar disorder (BD), quality of life (QOL) scores have been largely attributed to mood symptoms. However, impairments in QOL may occur even in euthymia, and differential factors have been put forward as important determinants of QOL. Our study was designed to assess the role of cognitive performance in self-reported QOL in patients with BD. In this cross-sectional study, we examined the relation between cognitive variables and self-reported QOL in 55 bipolar I euthymic patients and 50 healthy subjects. Participants were administered the World Health Organization Quality of Life Assessment--Abbreviated version and a battery of neuropsychological tests. BD patients showed lower scores in all QOL domains as compared with control subjects. Poorer self-reported QOL correlated significantly with worse cognitive performance, especially on tests of executive functioning and verbal abstraction. A linear regression model revealed that all QOL domains were significantly predicted by cognitive variables, with variances ranging from 12% to 37%, and from 24% to 54% when clinical variables were added to the model. Deficits in executive functioning and verbal abstraction were strong predictors of poor self-reported QOL. Our findings suggest that, along with mood stabilization, adequate cognitive functioning is desirable for achieving better QOL. These findings suggest that cognitive rehabilitation may be an important factor for restoring QOL to baseline levels among BD patients.


The effects of peppermint (Mentha piperita L.) and chicory (Cichorium intybus L.) in comparison with a prebiotic on productive performance, blood constituents, immunity and intestinal microflora in broiler chickens.
A total of 320 one-day-old broiler chickens were used in a 42-day feeding trial to evaluate the effects of peppermint (Mentha piperita L.) and chicory (Cichorium intybus L.) in comparison with a prebiotic on-growth performance, blood constitutes, immunity and intestinal microflora. The dietary treatments were as follows: basal diet (control); control + prebiotic (Fermacto™); control + 0.1% peppermint; control + 0.1% chicory, respectively. A significant (p < 0.05) body weight gain and feed intake was found at 21 and 42 days of growth period in broilers fed diet supplemented with 0.1% chicory compared with other groups. Feeding of prebiotic or chicory led to higher (p < 0.05) feed intake. Chickens fed control diet had higher (p < 0.05) abdominal fat compared with the other groups. Serum blood constituents indicated that broilers fed prebiotic or supplemented with peppermint or chicory had reduced (p < 0.05) levels of cholesterol, triglycerides and low-density lipoprotein than control group. Immunity-related parameters showed that chicken fed chicory had lower (p < 0.05) heterophil-to-lymphocyte ratio compared with the other groups. Intestinal microflora revealed that chickens fed prebiotic or herbals had higher count of Lactobacillus and lower E. coli than control. Thus, it can be concluded that broiler dietary supplementation with prebiotic or chicory can improve performance supporting positively health status.


Histone deacetylases in Trypanosoma brucei: two are essential and another is required for normal cell cycle progression.
Reversible protein acetylation is established as a modification of major regulatory significance. In particular, histone acetylation regulates access to genetic information in eukaryotes. For example, class I and class II histone deacetylases are regulatory components of corepressor complexes involved in cell cycle progression and differentiation. Here, we have investigated the function of such enzymes in Trypanosoma brucei, mono-flagellated parasitic protozoa that branched very early from the eukaryotic lineage. Four T. brucei genes encoding histone deacetylase orthologues have been identified, cloned and characterized. The predicted deacetylases, DAC1-4 are approximately 43, 61, 75 and 64 kDa respectively. They share significant similarity with mammalian and yeast class I (DAC1 and DAC2) and class II (DAC3 and DAC4) histone deacetylases, and all except DAC2 have the critical residues predicted to be required for deacetylase activity. In gene targeting experiments, DAC1 and DAC3 appear to be essential whereas DAC2 and DAC4 are not required for viability. Of the two mutant cell types, the dac4 mutant displays a delay in the G2/M phase of the cell cycle. Our results provide genetic validation of DAC1 and DAC3 as potential chemotherapy targets and demonstrate that T. brucei expresses at least three probable histone deacetylases with distinct function.


Cell culture assay for transient replication of human and animal papillomaviruses.
This unit contains protocols for evaluation of replication functionality of papillomavirus genomes or subgenomic fragments. Replication is measured after transient cotransfection of the genome (or subgenomic fragment) with expression vectors encoding the viral E1 and E2 proteins. Input DNA is methylated at the adenine of GATC sequences by propagation in E. coli. DNA that replicates in mammalian cells will lose the adenine methylation and become DpnI-resistant, while residual methylated input DNA will remain DpnI-sensitive. After transfection, DNA extraction, and DpnI digestion, replicated DNA can be detected by Southern blotting as a full-length plasmid, since it is resistant to digestion. This assay can be used to map the genomic location of a functional origin or to evaluate replication activity of mutations in either the origin DNA sequences or the E1 or E2 proteins.


Annular alpha-synuclein species from purified multiple system atrophy inclusions.
Oligodendroglial cytoplasmic inclusions composed of alpha-synuclein filamentous aggregates are the pathological hallmark of multiple system atrophy (MSA). We found that cortical tissue from MSA cases contains increased detergent-resistant high-molecular-weight alpha-synuclein species. To analyse these species, we immunopurified alpha-synuclein aggregates from pathological samples and examined their ultrastructures using scanning electron and atomic force microscopies. Purified aggregates consisted of bundles of filaments. After treatment with 1% sarcosine or 2% 3-[(3-cholamidopropyl) dimethyl-ammonio]-1-propanesulfonate (CHAPS) detergents, we observed frequent 30-50 nm annular particles, probably released from pathological aggregates due to the dissociation of filaments by the detergents. Antibody recognition imaging using a specific anti-alpha-synuclein antibody confirmed that the annular structures were positive for alpha-synuclein. In contrast to pathological alpha-synuclein, detergent treatment of recombinant alpha-synuclein yielded only smaller, 10-18 nm spherical particles. Our results demonstrate that detergent treatment of pathological MSA alpha-synuclein aggregates, but not recombinant alpha-synuclein, yields discrete alpha-synuclein-positive species with annular morphologies. The ability of the pathological alpha-synuclein to form annular aggregates may be an important factor contributing to the toxicity of the protein in disease that may have implications in designing therapeutic strategies aimed at detoxifying alpha-synuclein aggregates.


Impact of postal and telephone reminders on pick-up rates of unclaimed e-prescriptions.
To investigate the impact of a reminder (i.e., a mailed letter or short telephone call) from the pharmacy to patients, compared with no reminder in a control group, on the pick-up rates of unclaimed e-prescriptions. Patients, with e-prescriptions transmitted to four large community pharmacies in two counties in northern Sweden and remaining unclaimed after 4 weekdays, were randomised to one of two intervention groups (a mailed reminder or a short telephonic reminder) or a control group. Rates of patients' pick-up of their e-prescriptions at follow-up after about 1, 2 and 3 weeks. Altogether, 320 patients with e-prescriptions, transmitted from March 21 through April 6 and not picked-up or dispensed, were identified and randomised to the study. There were no statistically significant differences in overall pick-up rates between the groups or with respect to gender. However, pick-up rates increased with increasing age. Higher pick-up rates were observed for two subgroups (but only in the mailed reminder group compared with controls)--for cardiovascular drugs to men and for respiratory drugs to adolescents and young adults. A reminder (i.e., a mailed letter or short telephone call) from the pharmacy to the patient had no statistically significant effect on overall pick-up rates of unclaimed e-prescriptions compared with no reminders.


Time to coronary catheterization in patients with non-ST-segment elevation acute coronary syndrome and high Global Registry of Acute Coronary Events score.
Current guidelines recommend an early (<24 h) invasive coronary angiography (ICA) strategy in non-ST-segment elevation acute coronary syndrome (NSTE-ACS) patients with Global Registry of Acute Coronary Events (GRACE) score over 140. Evidence for this recommendation is based on older trials. Between 1 February 2016 and 31 July 2021, 1767 patients with a primary diagnosis of NSTE-ACS without indication for urgent ICA underwent ICA during index hospitalization. Six hundred and fifty-five patients underwent early invasive ICA (within 24 h) and 1112 underwent late ICA (between 24 h and 1 week). One hundred and seven patients had a GRACE risk score of 140 or above and 1660 had a GRACE risk score under 140. The primary composite outcome was all-cause mortality, stroke, and recurrent myocardial infarction (MI). Median time from admission to ICA was 13.3 h (IQR 6.0-20.6) for the early group and 59.9 h for the late group (IQR 23.5-96.3). There was no difference between the early and late ICA groups in the primary composite outcome [late catheterization >24 h hazard ratio 1.196, 95% confidence interval (CI) 0.969-1.475, P-value 0.096]. A multivariable Cox regression model for the composite outcome revealed no difference between the early and late ICA groups (late catheterization >24 h hazard ratio 1.0735, 95% CI 0.862-1.327, P-value 0.512) with no effect for performing early ICA in patients with GRACE score over 140 (hazard ratio 1.291, 95% CI 0.910-1.831, P-value 0.151). An early ICA strategy in patients with NSTE-ACS patients and GRACE risk score over 140, compared with late ICA, was not associated with improved composite outcome of death, myocardial infarction, and stroke at 1 year.


Fluid dynamics in a full-scale flat sheet MBR, an experimental and numerical study.
Fluid dynamics is used for fouling mitigation in membrane bioreactors (MBRs), whereby a proper understanding of the fluid dynamics is of great interest. The influence of fluid dynamics has led to the use of computational fluid dynamics for optimizing MBR systems. In this work, a model has been validated for flat sheet membranes, with use of the Eulerian multiphase method. The model is validated against a comparable setup where the liquid velocities are measured with a laser Doppler anemometer (LDA). Furthermore, the Eulerian multiphase approach is validated against the more numerical direct volume of fluid (VOF) approach with sludge properties for the liquid, resulting in an error between the models of less than 2% for the wall shear stresses. The VOF model further showed that the horizontal components contribute significantly to the total wall shear stresses. The model has been applied to a full-scale setup for studying the effect of deflecting membranes as deflections have been seen in production. Minimizing the deflection of the membrane sheets was crucial to achieve a good operating condition as a deflection of 2 mm in a setup with a gap of 7 mm decreased the wall shear stresses with as much as 40% on average on the specific membrane surface.


Clinical perspectives on hereditary hemochromatosis.
Hereditary hemochromatosis (HH) comprises a group of inherited disorders of iron metabolism that can result in progressive iron overload, morbidity, and mortality, generally in adulthood. HFE-related HH is the most common type of HH and will form the core of this discussion. The discovery of new proteins and gene mutations has defined other types of HH, termed non-HFE HH. The regulatory protein hepcidin has a central role in iron homeostasis in these disorders. While the liver is the predominant organ of iron deposition and iron-overload-related disease in HFE-related HH, involvement of extrahepatic tissue can also result in morbidity and mortality if the disorder is not diagnosed before organ damage develops. This review traverses the road from HFE genotype to phenotype with a focus on clinical penetrance, modifier factors for disease expression, and current thoughts and controversies on HH diagnosis and screening.


Cephalosporium maydis is a distinct species in the Gaeumannomyces-Harpophora species complex.
Cephalosporium maydis is an important plant pathogen whose phylogenetic position relative to other fungi has not been established clearly. We compared strains of C. maydis, strains from several other plant-pathogenic Cephalosporium spp. and several possible relatives within the Gaeumannomyces-Harpophora species complex, to which C. maydis has been suggested to belong based on previous preliminary DNA sequence analyses. DNA sequences of the nuclear genes encoding the rDNA ITS region, β-tubulin, histone H3, and MAT-2 support the hypothesis that C. maydis is a distinct taxon within the Gaeumannomyces-Harpophora species complex. Based on amplified fragment length polymorphism (AFLP) profiles, C. maydis also is distinct from the other tested species of Cephalosporium, Phialophora sensu lato and members of Gaeumannomyces-Harpophora species complex, which supports its classification as Harpophora maydis. Oligonucleotide primers for H. maydis were developed that can be used in a PCR diagnostic protocol to rapidly and reliably detect and identify this pathogen. These diagnostic PCR primers will aid the detection of H. maydis in diseased maize because this fungus can be difficult to detect and isolate, and the movement of authentic cultures may be limited by quarantine restrictions.


Image Processing in Cryo-Electron Microscopy of Single Particles: The Power of Combining Methods.
Cryo-electron microscopy has established as a mature structural biology technique to elucidate the three-dimensional structure of biological macromolecules. The Coulomb potential of the sample is imaged by an electron beam, and fast semi-conductor detectors produce movies of the sample under study. These movies have to be further processed by a whole pipeline of image-processing algorithms that produce the final structure of the macromolecule. In this chapter, we illustrate this whole processing pipeline putting in value the strength of "meta algorithms," which are the combination of several algorithms, each one with different mathematical rationale, in order to distinguish correctly from incorrectly estimated parameters. We show how this strategy leads to superior performance of the whole pipeline as well as more confident assessments about the reconstructed structures. The "meta algorithms" strategy is common to many fields and, in particular, it has provided excellent results in bioinformatics. We illustrate this combination using the workflow engine, Scipion.


Nephrotoxicity-induced proteinuria increases biomarker diagnostic thresholds in acute kidney injury.
Paraquat ingestion is frequently fatal. While biomarkers of kidney damage increase during paraquat-induced acute kidney injury (AKI), significant concurrent proteinuria may alter diagnostic thresholds for diagnosis and prognosis to an unknown extent. This study evaluated the effect of albuminuria on biomarker cutoffs for diagnosis and outcome prediction. This was a multi-centre prospective clinical study of patients following acute paraquat self-poisoning in 5 Sri Lankan hospitals. Biomarker concentrations were quantified using ELISA and microbead assays and correlated with urinary albumin. Functional-AKI was defined by the Acute Kidney Injury Network serum creatinine definition and alternatively by a ≥50% increase in serum cystatin C. Albuminuria was defined as albumin-creatinine ratio >30 mg/g. The study outcomes were compared with a retrospective analysis of a pre-clinical study of paraquat-induced nephrotoxicity with appropriate controls. Albuminuria was detected in 34 of 50 patients, and increased with functional-AKI severity. The concentrations of uNGAL, uCysC, uClusterin, uβ2M, and uKIM-1 were higher in albuminuric compared to non-albuminuric patients (p < 0.001). Albuminuria correlated with biomarker concentration (r > 0.6, p < 0.01) and was associated with death (p = 0.006). Optimal biomarker cutoffs for prediction of death were higher in the albuminuric group. Similar outcomes with more detailed analysis were obtained in experimental paraquat nephrotoxicity. Albuminuria was associated with paraquat-induced nephrotoxicity and increased excretion of low-molecular weight protein biomarkers. AKI biomarker cutoffs for diagnosis, outcome prediction and AKI stratification increased in the presence of albuminuria. This may lead to over-diagnosis of AKI in conditions independently associated with proteinuria.


Evaluation of the effect of using mobile phones on male fertility.
The problem of the lack of offspring is a phenomenon concerning approximately 15% of married couples in Poland. Infertility is defined as inability to conceive after a year of sexual intercourses without the use of contraceptives. In half of the cases the causative factor is the male. Males are exposed to the effect of various environmental factors, which may decrease their reproductive capabilities. A decrease in male fertility is a phenomenon which occurs within years, which may suggest that one of the reasons for the decrease in semen parameters is the effect of the development of techniques in the surrounding environment. A hazardous effect on male fertility may be manifested by a decrease in the amount of sperm cells, disorders in their mobility, as well as structure. The causative agents may be chemical substances, ionizing radiation, stress, as well as electromagnetic waves. The objective of the study was the determination of the effect of the usage of cellular phones on the fertility of males subjected to marital infertility therapy. The following groups were selected from among 304 males covered by the study: Group A: 99 patients who did not use mobile phones, Group B: 157 males who have used GSM equipment sporadically for the period of 1-2 years, and Group C: 48 people who have been regularly using mobile phone for more than 2 years. In the analysis of the effect of GSM equipment on the semen it was noted that an increase in the percentage of sperm cells of abnormal morphology is associated with the duration of exposure to the waves emitted by the GSM phone. It was also confirmed that a decrease in the percentage of sperm cells in vital progressing motility in the semen is correlated with the frequency of using mobile phones.


Molecular diagnosis of congenital adrenal hyperplasia due to 21-hydroxylase deficiency: an update of new CYP21A2 mutations.
Steroid 21-hydroxylase deficiency is present in more than 90% of patients with congenital adrenal hyperplasia, an inherited metabolic disorder of adrenal steroidogenesis. Impaired enzymatic activity leads to the accumulation of metabolic intermediates (progesterone and 17-hydroxyprogesterone), which results in excessive androgen production and varied signs of virilisation. CYP21A2 is an active gene and encodes for the steroid 21-hydroxylase enzyme, whereas CYP21A1P is an inactive pseudogene that contains a series of deleterious mutations. The major part of disease-causing mutations in CYP21A2 alleles are CYP21A1P-derived sequence transferred to the active gene by macro or microconversion events. Approximately 5% of all disease-causing CYP21A2 alleles harbour rare mutations that do not originate from the pseudogene. A list of all reported CYP21A2 mutations can be found in the CYP21A2 database created by the Human Cytochrome P450 (CYP) Allele Nomenclature Committee (http:www.imm.Ki.se/CYPalleles/cyp21.htm). Unfortunately, the last update of this database was in 2006. However, over the last 4 years many other novel CYP21A2 mutations have been reported in PubMed. The aim of this review is to provide a focus on the molecular and genetic aspects of the diagnosis of 21-hydroxylase deficiency. In addition, an updated list of the last new CYP21A2 mutations is included.


Study of shear properties and shear model of soil-rock mixture.
This study investigates the shear characteristics of soil-rock mixtures, a critical factor influencing slope stability in engineering construction. Soil-rock mixtures, often exhibit poor integrity and can easily soften in water due to geological influences. The YT1200 direct shear drawing friction system was employed to conduct shear tests, analyzing the effect of varying water content and fine particle mass fraction under different normal stresses. Utilizing fractional derivatives, we formulated a fractional derivative shear model. Test results illustrated a softening phenomenon post achieving peak shear stress in the soil-rock mixture. It was found that peak shear stress is directly proportional to the normal stress, and inversely proportional to both water content and fine particle mass fraction. Additionally, the cohesion and internal friction angle decrease according to a power function with increasing water content, and non-linearly decrease with the rise of fine particle mass fraction. The proposed shear model aptly simulates the entire shear failure process of the soil-rock mixture, effectively analyzing the influence of key factors on shear characteristics. These findings contribute to the strength prediction and numerical simulation of soil-rock mixtures, thereby aiding in the design of reinforcement schemes and slope stability analysis.


Optimizing the dose of glutamine dipeptides and antioxidants in critically ill patients: a phase I dose-finding study.
Supplementation with glutamine and antioxidants may be associated with an improvement in clinical outcomes, but the optimal dose of these substrates is unknown. The purpose of this study was to determine the safety of high doses of glutamine combined with antioxidants in critically ill patients. We conducted a single-center, open-label, dose-escalating clinical trial. Mechanically ventilated adult patients with clinical evidence of hypoperfusion were sequentially enrolled to 1 of 5 groups. Group 1 (n = 30): no supplementation; group 2 (n = 7): 0.35 g/kg/d of glutamine IV; group 3 (n = 7): same as group 2 plus 15 g/d of glutamine and 150 microg of selenium enterally; group 4 (n = 7): same as group 2 plus 30 g/d of glutamine and 300 microg of selenium enterally; and group 5 (n = 7): same as group 4 plus an additional 500 microg of selenium IV. After enrollment, nutrients were started as soon as possible. All patients were fed enterally according to clinical practice guidelines. The primary outcomes for this study were change in sequential organ function assessment (SOFA) score and safety parameters. Secondary outcomes included whole blood glutathione (GSH), thiobarbituric acid reactive substances (TBARS), and blood cells' mitochondrial DNA/nuclear DNA ratio (RATIO). There were no adverse events attributable to the study nutrients, and the maximum and Delta SOFA did not differ across groups. In group 2, a significant decrease in GSH levels was observed (p = .03). With subsequent groups, the slopes straighten out and the p values are no longer significant, suggesting a greater preservation of GSH levels with escalating doses. In group 2, the slope of the line representing TBARS was horizontal. With subsequent groups, the slopes decrease, and by group 5, this decrease reaches statistical significance (p = .03), suggesting a greater reduction in oxidative stress with the higher doses in group 5. The difference in slopes across all groups describing the mitochondrial RATIO is statistically significant (p = .001), again suggesting that, with higher doses, there is increased mitochondrial function. The doses of glutamine and antioxidants tested in this study seem to be safe and may have positive effects on some mechanistic endpoints. A larger trial will be necessary to confirm their therapeutic effects.


Adsorptive endocytosis and membrane recycling by cultured primary bovine brain microvessel endothelial cell monolayers.
The dynamics of membrane recycling were examined in primary cultures of brain microvessel endothelial cells (BMECs). Because the BMEC surface was dominated by galactosylated glycoconjugates, ricin agglutinin (RCAI) was used as a tracer to follow the endocytosis and recycling of RCAI binding sites. These binding sites accounted for 75% of the iodinatable or most externally disposed plasma membrane proteins. Because greater than 90% of the RCAI that had bound to BMECs was removed by a brief, nontoxic treatment with galactose, the amounts and kinetics for internalization and efflux of [125I]RCAI were measured. Both endocytosis and efflux were energy dependent. By using pseudo-first-order kinetics, the t1/2 values for RCAI binding, internalization and efflux were 5, 18 and 13-14 min, respectively. By comparing efflux with and without galactose present, we found that 60% of the RCAI binding sites that had been internalized were returned to the cell surface and reinternalized. Quantifying the distribution of gold-RCAI following internalization showed kinetics consistent with that obtained using radiolabeled RCAI. Both horseradish peroxidase (HRP) and gold-conjugated RCAI that had bound BMEC at 4 degrees C became localized within more caveolae within 2.5 min of warming to 37 degrees C to permit endocytosis. With time, RCAI appeared within endosomes and tubules and vesicles of which some were located in the trans-Golgi network (TGN). The distribution of HRP-RCAI contrasted with that of free HRP, which was not routed to the TGN. The absence of RCAI conjugates in association with the basolateral membrane domain suggested the presence of functional tight junctions and maintenance of polarity throughout the duration of these experiments. These results showed that membrane recycling was more extensive and much slower than fluid-phase endocytosis in cultured BMECs. Moreover, we found that endocytosis of membrane by BMECs in culture was similar to that reported for brain endothelium in vivo in that a fraction of the cell surface membrane was routed to the TGN.


In vitro production of cat-restricted Toxoplasma pre-sexual stages.
Sexual reproduction of Toxoplasma gondii, confined to the felid gut, remains largely uncharted owing to ethical concerns regarding the use of cats as model organisms. Chromatin modifiers dictate the developmental fate of the parasite during its multistage life cycle, but their targeting to stage-specific cistromes is poorly described1,2. Here we found that the transcription factors AP2XII-1 and AP2XI-2 operate during the tachyzoite stage, a hallmark of acute toxoplasmosis, to silence genes necessary for merozoites, a developmental stage critical for subsequent sexual commitment and transmission to the next host, including humans. Their conditional and simultaneous depletion leads to a marked change in the transcriptional program, promoting a full transition from tachyzoites to merozoites. These in vitro-cultured pre-gametes have unique protein markers and undergo typical asexual endopolygenic division cycles. In tachyzoites, AP2XII-1 and AP2XI-2 bind DNA as heterodimers at merozoite promoters and recruit MORC and HDAC3 (ref. 1), thereby limiting chromatin accessibility and transcription. Consequently, the commitment to merogony stems from a profound epigenetic rewiring orchestrated by AP2XII-1 and AP2XI-2. Successful production of merozoites in vitro paves the way for future studies on Toxoplasma sexual development without the need for cat infections and holds promise for the development of therapies to prevent parasite transmission.


Functional chromophores synthesized via multicomponent Reactions: A review on their use as cell-imaging probes.
This review aims to provide a comprehensive overview of recent advancements and applications of fluorescence imaging probes synthesized via MCRs (multicomponent reactions). These probes, also known as functional chromophores, belong to a currently investigated class of fluorophores that are presently being successfully applied in bioimaging experiments, especially in various living cell lineages. We describe some of the MCRs that have been employed in the synthesis of these probes and explore their applications in biological imaging, with an emphasis on cellular imaging. The review also discusses the challenges and future perspectives in the field, particularly considering the potential impact of MCR-based fluorescence imaging probes on advancing this field of research in the coming years. Considering that this area of research is relatively new and nearly a decade has passed since the first publication, this review also provides a historical perspective on this class of fluorophores, highlighting the pioneering works published between 2011 and 2016.


Genomic and phenotypic characterization of a wild medaka population: towards the establishment of an isogenic population genetic resource in fish.
Oryzias latipes (medaka) has been established as a vertebrate genetic model for more than a century and recently has been rediscovered outside its native Japan. The power of new sequencing methods now makes it possible to reinvigorate medaka genetics, in particular by establishing a near-isogenic panel derived from a single wild population. Here we characterize the genomes of wild medaka catches obtained from a single Southern Japanese population in Kiyosu as a precursor for the establishment of a near-isogenic panel of wild lines. The population is free of significant detrimental population structure and has advantageous linkage disequilibrium properties suitable for the establishment of the proposed panel. Analysis of morphometric traits in five representative inbred strains suggests phenotypic mapping will be feasible in the panel. In addition, high-throughput genome sequencing of these medaka strains confirms their evolutionary relationships on lines of geographic separation and provides further evidence that there has been little significant interbreeding between the Southern and Northern medaka population since the Southern/Northern population split. The sequence data suggest that the Southern Japanese medaka existed as a larger older population that went through a relatively recent bottleneck approximately 10,000 years ago. In addition, we detect patterns of recent positive selection in the Southern population. These data indicate that the genetic structure of the Kiyosu medaka samples is suitable for the establishment of a vertebrate near-isogenic panel and therefore inbreeding of 200 lines based on this population has commenced. Progress of this project can be tracked at http://www.ebi.ac.uk/birney-srv/medaka-ref-panel.


Predictors of unprotected sex among female sex workers in Madagascar: comparing semen biomarkers and self-reported data.
Research on the determinants of condom use and condom non-use generally has relied on self-reported data with questionable validity. We identified predictors of recent, unprotected sex among 331 female sex workers in Madagascar using two outcome measures: self-reports of unprotected sex within the past 48 h and detection of prostate-specific antigen (PSA), a biological marker of recent semen exposure. Multivariable logistic regression revealed that self-reported unprotected sex was associated with three factors: younger age, having a sipa (emotional partner) in the prior seven days, and no current use of hormonal contraception. The sole factor related to having PSA detected was prevalent chlamydial infection (adjusted odds ratio, 4.5; 95% confidence interval, 2.0-10.1). Differences in predictors identified suggest that determinants of unprotected sex, based on self-reported behaviors, might not correlate well with risk of semen exposure. Caution must be taken when interpreting self-reported sexual behavior measures or when adjusting for them in analyses evaluating interventions for the prevention of HIV/STIs.


Traceless Protein-Selective Glycan Labeling and Chemical Modification.
Executing glycan editing at a molecular level not only is pivotal for the elucidation of complicated mechanisms involved in glycan-relevant biological processes but also provides a promising solution to potentiate disease therapy. However, the precision control of glycan modification or glyco-editing on a selected glycoprotein is by far a grand challenge. Of note is to preserve the intact cellular glycan landscape, which is preserved after editing events are completed. We report herein a versatile, traceless glycan modification methodology for customizing the glycoforms of targeted proteins (subtypes), by orchestrating chemical- and photoregulation in a protein-selective glycoenzymatic system. This method relies on a three-module, ligand-photocleavable linker-glycoenzyme (L-P-G) conjugate. We demonstrated that RGD- or synthetic carbohydrate ligand-containing conjugates (RPG and SPG) would not activate until after the ligand-receptor interaction is accomplished (chemical regulation). RPG and SPG can both release the glycoenzyme upon photoillumination (photoregulation). The adjustable glycoenzyme activity, combined with ligand recognition selectivity, minimizes unnecessary glycan editing perturbation, and photolytic cleavage enables precise temporal control of editing events. An altered target protein turnover and dimerization were observed in our system, emphasizing the significance of preserving the native physiological niche of a particular protein when precise modification on the carbohydrate epitope occurs.


Nonsolar energy use and one-hundred-year global warming potential of Iowa swine feedstuffs and feeding strategies.
Demand for nonsolar energy and concern about the implications of fossil fuel combustion have encouraged examination of energy use associated with agriculture. The United States is a global leader in pig production, and the United States swine industry is centered in Iowa. Feed is the largest individual input in pig production, but the energy consumption of the Iowa swine feed production chain has yet to be critically examined. This analysis examines nonsolar energy use and resulting 100-yr global warming potential (GWP) associated with the swine feed production chain, beginning with cultivation of crops and concluding with diet formulation. The nonsolar energy use and accompanying 100-yr GWP associated with production of 13 common swine feed ingredients are estimated. Two diet formulation strategies are considered for 4 crop sequence x ingredient choice combinations to generate 8 crop sequence x diet formulation scenarios. The first formulation strategy (simple) does not include synthetic AA or phytase. The second strategy (complex) reduces CP content of the diet by using L-lysine to meet standardized ileal digestibility lysine requirements of pigs and includes the exogenous enzyme phytase. Regardless of crop sequence x diet formulation scenario, including the enzyme phytase is energetically favorable and reduces the potential excretion of P by reducing or removing inorganic P from the complete diet. Including L-lysine reduces the CP content of the diet and requires less nonsolar energy to deliver adequate standardized ileal digestible lysine than simply feeding soybean meal. Replacing soybean meal with full-fat soybeans is not energetically beneficial under Iowa conditions. Swine diets including dried distillers grains with solubles and crude glycerol require approximately 50% more nonsolar energy inputs than corn-soybean meal diets or corn-soybean meal diets including oats. This study provides essential information on cultivation, processing, and manufacture of swine feed ingredients in Iowa that can be coupled with other models to estimate the nonsolar energy use and 100-yr GWP of pig production.


Measurement of ultrasonic pulse velocity with improved accuracy using automatic threshold error correction.
Ultrasonic Pulse Velocity (UPV) measurement is extensively used to monitor the strength and health of concrete structures as per American Society for Testing and Materials C 597 - 09. The commercially available UPV measurement systems work on the basis of single threshold detection of the received signal. Therefore, measurement accuracy is affected due to threshold error. The effect is sensitive to the signal amplitude reaching the threshold comparator and, hence, receiver gain. It is observed that a UPV tester operating at 50 kHz to test concrete might generate an error of up to 10% in the ultrasonic transit time measurement of 50 µs. Hence, it is of great concern and needs to be improved. In this article, the UPV measurement circuit capable of detecting and compensating the threshold error is described. The threshold error correction is achieved with the help of two threshold comparators and two hybrid counters. The circuit developed minimizes the threshold error for wide receiver gain. The measurement carried out with the developed system shows significant improvement, having deviations within 100 ns.


End-of-life after birth: death and dying in a neonatal intensive care unit.
In canonical modern bioethics, withholding and withdrawing medical interventions for dying patients are considered morally equivalent. However, electing not to administer cardiopulmonary resuscitation (CPR) struck us as easily distinguishable from withdrawing mechanical ventilation. Moreover, withdrawing mechanical ventilation from a moribund infant "feels" different from withdrawing mechanical ventilation from a hemodynamically stable child with a severe neurologic insult. Most previous descriptions of withdrawing and withholding intervention in the neonatal intensive care unit (NICU) have blurred many of these distinctions. We hypothesized that clarifying them would more accurately portray the process of end-of-life decision-making in the NICU. We reviewed the charts of all newborn infants who had birth weight >400 g and died in our hospital in 1988, 1993, and 1998 and extracted potential ethical issues (resuscitation, withdrawal, withholding, CPR, do-not-resuscitate orders, neurologic prognosis, ethics consult) surrounding each infant's death. Using traditional definitions, roughly half of all deaths in our NICU in 1993 and 1998 were associated with "withholding or withdrawing." In addition, by 1998, >40% of our NICU deaths could be labeled "active withdrawal," reflecting the extubation of infants regardless of their physiologic instability. This practice is growing over time. However, 2 important conclusions arise from our more richly elaborated descriptions of death in the NICU. First, when CPR was withheld, it most commonly occurred in the context of moribund infants who were already receiving ventilation and dopamine. Physiologically stable infants who were removed from mechanical ventilation for quality-of-life reasons accounted for only 3% of NICU deaths in 1988, 16% of NICU deaths in 1993, and 13% of NICU deaths in 1998. Moreover, virtually none of these active withdrawals took place in premature infants. Second, by 1998 infants, who died without CPR almost always had mechanical ventilation withdrawn. Finally, the median and average day of death for 100 nonsurvivors who received full intervention did not differ significantly from the 78 nonsurvivors for whom intervention was withheld. In our unit, a greater and greater percentage of doomed infants die without ever receiving chest compressions or epinephrine boluses. Rather, we have adopted a nuanced approach to withdrawing/withholding NICU intervention, providing what we hope is a humane approach to end-of-life decisions for doomed NICU infants. We suggest that ethical descriptions that reflect these nuances, distinguishing between withholding and withdrawing interventions from physiologically moribund infants or physiologically stable infants with morbid neurologic prognoses, provide a more accurate reflection of the circumstances of dying in the NICU.


TNF receptor type I-dependent activation of innate responses to reduce intestinal damage-associated mortality.
Ligation of tumor necrosis factor (TNF) receptors (TNFRs) with TNF plays a critical role in the pathogenesis of human inflammatory bowel disease (IBD). However, it remains unclear which cell types activated through TNFR-associated signaling cascades are involved in the pathogenesis of colitis. Recombination activating gene-1 (RAG) knockout (KO) (no T or B cells)-based TNFR double and triple KO mice were generated. Bone marrow (BM) chimera mice in which BM-derived myeloid cells, but not colonic epithelial cells (CECs), express TNFRs were also generated. Colitis was induced by administration of dextran sodium sulfate (DSS) in distilled water. Murine lines and chimeras were assessed for disease severity, histopathology, apoptotic cell rate, epithelial proliferation, and bacterial invasion rate. Following DSS administration, mice lacking both RAG and TNFR1 exhibited a high mortality (>80%) rate with an impaired CEC regeneration compared with RAG KO and RAG x TNFR2 double KO (DKO) mice. Transplantation of RAG KO-derived BM cells restored CEC regeneration and rescued the majority of recipient RAG x TNFR1 DKO mice from DSS-induced mortality. After BM transplantation, RAG x TNFR1 DKO mice exhibited an increased rate of apoptosis in the colonic lamina propria macrophages in association with the activation of caspases. In addition, BM reconstitution directly or indirectly enhanced the proliferation of CECs by activating mitogen-activated protein kinase and phosphoinositide-3 kinase/Akt pathways. TNFR1-signaling cascade in colonic myeloid lineage cells contributes to the suppression of acute damage-associated mortality presumably by controlling CEC homeostasis.


Acetylene reduction assays for nitrogen fixation in freshwaters: a note of caution.
Lake water samples were observed to transform [14-C]ethylene into water-soluble compounds that were undetectable by conventional acetylene reduction assay procedures. Methane oxidizing bacteria, which are known to be common in freshwaters, appeared to be responsible for this activity. As much as 28 percent of added ethylene has been observed to be transformed and this figure is probably an underestimate. It is suggested that acetylene reduction assays may not be accurately applied to samples containing methane oxidizing bacteria.


NUCLEAR SEGMENTATION IN MICROSCOPE CELL IMAGES: A HAND-SEGMENTED DATASET AND COMPARISON OF ALGORITHMS.
Image segmentation is an essential step in many image analysis pipelines and many algorithms have been proposed to solve this problem. However, they are often evaluated subjectively or based on a small number of examples. To fill this gap, we hand-segmented a set of 97 fluorescence microscopy images (a total of 4009 cells) and objectively evaluated some previously proposed segmentation algorithms.We focus on algorithms appropriate for high-throughput settings, where only minimal user intervention is feasible.The hand-labeled dataset (and all software used to compare methods) is publicly available to enable others to use it as a benchmark for newly proposed algorithms.


A 64-Year-Old Man with Low Back Pain Due to Clostridium perfringens Lumbar Discitis.
BACKGROUND Lumbar discitis caused by Clostridium perfringens is extremely rare. There have only been 7 published cases of confirmed discitis caused by Clostridium perfringens. We write this report to underscore this unusual relationship by discussing an additional case and providing a review of the previously published cases so clinicians can adequately evaluate and treat patients presenting with discitis. CASE REPORT A 64-year-old morbidly obese man presented with an acute onset of worsening back pain and generalized weakness after incurring physical trauma related to falling. Additionally, he also developed fever and chills before the presentation. Based on the clinical presentation and elevated serum levels of inflammatory markers, magnetic resonance imaging was ordered, which showed L5-S1 discitis with extension of infection into the epidural space. Fluoroscopy-guided aspiration of the L5-S1 epidural space facilitated the detection of Clostridium perfringens as the involved pathogen. Based on the antibiotic susceptibility report, the patient was treated with intravenous ampicillin for 8 weeks, after which his symptoms resolved. CONCLUSIONS Diagnosis of discitis can be very challenging due to its ambiguous clinical presentation, especially in the elderly population due to the presence of underlying degenerative changes. Even though Clostridium perfringens remains a rare cause of lumbar discitis, it should be considered as a pathogen capable of causing infection of the vertebrae and intervertebral discs, thus allowing clinicians to make necessary diagnostic evaluations to provide appropriate targeted treatment to patients presenting with discitis.


A Longitudinal Study of Local Gyrification Index in Young Boys With Autism Spectrum Disorder.
Local gyrification index (LGI), a metric quantifying cortical folding, was evaluated in 105 boys with autism spectrum disorder (ASD) and 49 typically developing (TD) boys at 3 and 5 years-of-age. At 3 years-of-age, boys with ASD had reduced gyrification in the fusiform gyrus compared with TD boys. A longitudinal evaluation from 3 to 5 years revealed that while TD boys had stable/decreasing LGI, boys with ASD had increasing LGI in right inferior temporal gyrus, right inferior frontal gyrus, right inferior parietal lobule, and stable LGI in left lingual gyrus. LGI was also examined in a previously defined neurophenotype of boys with ASD and disproportionate megalencephaly. At 3 years-of-age, this subgroup exhibited increased LGI in right dorsomedial prefrontal cortex, cingulate cortex, and paracentral cortex, and left cingulate cortex and superior frontal gyrus relative to TD boys and increased LGI in right paracentral lobule and parahippocampal gyrus, and left precentral gyrus compared with boys with ASD and normal brain size. In summary, this study identified alterations in the pattern and development of LGI during early childhood in ASD. Distinct patterns of alterations in subgroups of boys with ASD suggests that multiple neurophenotypes exist and boys with ASD and disproportionate megalencephaly should be evaluated separately.


[Intracellular distribution of a DNA and carminomycin complex].
The complex of the tritium labeled DNA with carminomycin was injected intravenously to mice. Five h after the complex injection the rats were sacrificed. The liver was isolated and homogenized. The cells were consecutively divided into fractions: intercellular liquid, cytoplasm, nuclear juice and chromatin. The content of high-molecular exogenous DNA and the products of its degradation, as well as the quantity of free and DNA bound carminomycin were determined radiometrically and spectrofluorometrically in every of the above fractions. It was found that the carminomycin complex located in the liver remained unchanged for a long time after injection and penetrated into the cell in the form of the complex. In the cytoplasm, the complex disintegrated to liberate carminomycin which penetrated into the nucleus where it was bound to chromatin DNA.


A Secure and Decentralized Trust Management Scheme for Smart Health Systems.
The Internet of Things (IoT) growth is extremely fast and it now has found its way to healthcare applications too. Many smart health gadgets and devices are helping practitioners in collecting medical information and monitoring patients. In this distributed system, information or service is sometimes shared and used by other devices. Considering the importance of health-related information and the decisions made based on it, there should be some sort of assurance on the security and quality of the services or information provided. Trust management is an efficient means of promoting application security and reliability in these cases. However, due to some limitations that are specific to IoT, traditional trust evaluation algorithms cannot be employed or do not yield satisfactory results. In this paper, evidence theory is exploited to design a decentralized service-oriented trust management model for healthcare IoT. A measure of evidence distance is used to reward well-behaving healthcare service/information providers as well as referrers and punish malicious entities. In this context-aware model, trust is estimated based on direct experiences and indirect feedbacks of recommenders. The process runs in two contexts; trust to healthcare service and trust to recommendation. When personal direct experience does not exist, trust to a source or service is estimated by applying the combinatorial laws of evidence theory and integrating indirect trust values. The proposed model is secure against bad-mouthing, good-mouthing, and on-off attacks due to its dynamic parameters and using the concept of evidence distance. Our results confirm the robustness and efficiency of this scheme.


Human Tissue-Resident Memory T Cells Are Defined by Core Transcriptional and Functional Signatures in Lymphoid and Mucosal Sites.
Tissue-resident memory T cells (TRMs) in mice mediate optimal protective immunity to infection and vaccination, while in humans, the existence and properties of TRMs remain unclear. Here, we use a unique human tissue resource to determine whether human tissue memory T cells constitute a distinct subset in diverse mucosal and lymphoid tissues. We identify a core transcriptional profile within the CD69+ subset of memory CD4+ and CD8+ T cells in lung and spleen that is distinct from that of CD69- TEM cells in tissues and circulation and defines human TRMs based on homology to the transcriptional profile of mouse CD8+ TRMs. Human TRMs in diverse sites exhibit increased expression of adhesion and inhibitory molecules, produce both pro-inflammatory and regulatory cytokines, and have reduced turnover compared with circulating TEM, suggesting unique adaptations for in situ immunity. Together, our results provide a unifying signature for human TRM and a blueprint for designing tissue-targeted immunotherapies.


Barriers to the use of toxicogenomics data in human health risk assessment: A survey of Canadian risk assessors.
Regulatory agencies worldwide need to modernize human health risk assessment (HHRA) to meet challenges of the 21st century. Toxicogenomics is at the core of this improvement. Today, however, the use of toxicogenomics data in HHRA is very limited. The purpose of this survey was to identify barriers to the application of toxicogenomics data in HHRA by human health risk assessors. An online survey targeting Canadian risk assessors gathered information on their knowledge and perception of toxicogenomics, their current and future inclusion of toxicogenomics data in HHRA, and barriers to the use of such data. Twenty-nine (29) participants completed a questionnaire after 2 months of solicitation. The results show that the application of toxicogenomics data in Canada is marginal, with 85% of respondents reporting that they never or rarely used such data. Knowledge of toxicogenomics by Canadian risk assessors is also limited: about two-thirds of respondents (68%) were not at all or only slightly familiar with the concept. Lack of guidelines for toxicogenomics data interpretation, data quality assessment and on their use in HHRA, were found to be major barriers. In conclusion, there is a need for interventions aimed at facilitating the use of toxicogenomics data in HHRA, when available.


Exploring the Value of Using Color-Coded Quantitative DSA Evaluation on Bilateral Common Carotid Arteries in Predicting the Reliability of Intra-Ascending Aorta Flat Detector CT-CBV Maps.
Cerebral blood volume, acquired with flat panel detector CT by injecting contrast medium into the ascending aorta, enabled real-time acquisition of brain functional information with remarkable reduction of contrast medium usage comparing to an intravenous injection approach. However, individual vasculature and flow variations cause inhomogeneous contrast medium distribution and unexpected asymmetric perfusion for certain patients even without cerebral circulatory disorders. This work aimed at testing the feasibility of using color-coded quantitative DSA to predict the reliability of flat panel detector CT-based CBV maps by injecting contrast medium into the ascending aorta by exploring the correlation between measurements of color-coded quantitative DSA and the symmetry of CBV maps. For 12 patients without perfusion-related cerebral abnormities, color-coded quantitative DSA at the aortic arch and flat panel detector CT-based CBV maps by injecting contrast medium into the ascending aorta were acquired. In color-coded quantitative DSA, ROIs were defined on the bilateral common carotid arteries. Time-density curves were extracted, and area under the curve values were calculated. To evaluate brain perfusion symmetry, we defined ROIs on the anterior and middle cerebral artery territories in CBV maps, and quantitative CBV values were extracted. Eight patients demonstrated good perfusion symmetry with relative CBV of 0.96 ± 0.06, and their relative area under the curve was found to be 0.99 ± 0.02. For the other 4 patients, CBV from the left hemisphere was significantly lower than that from the right with relative CBV of 0.81 ± 0.09. This asymmetric perfusion was confirmed by the color-coded quantitative DSA with relative area under the curve values of 0.79 ± 0.03. This preliminary study showed good correlation between relative area under the curve from color-coded quantitative DSA and relative CBV from CBV maps. Color-coded quantitative DSA potentially helped sort out patients whose vascular anatomy could support reliable CBV acquisitions of flat detector CT by injecting contrast medium into the ascending aorta.


Role of Oxidative Stress and Reduced Endogenous Hydrogen Sulfide in Diabetic Nephropathy.
Persistent hyperglycemia lead towards depletion of hydrogen sulfide (H2S) resulting in generation of oxidative stress and diabetic nephropathy. The aim of the current study was to explore the antioxidant potential of H2S and captopril, a -SH containing compound in streptozotocin (STZ)-induced diabetic nephropathy. Fifty four Wistar-Kyoto (WKY) rats male (200-250g) were divided into nine groups (n=6) with each group injected once with STZ (60mg/kg i.p) except normal control. After 3 weeks of induction of diabetes, groups were assigned as normal control, diabetic control, diabetic-captopril, diabetic-NaHS, diabetic-captopril-NaHS, diabetic-spironolactone, diabetic-metformin, diabetic-metformin-NaHS and diabetic-vitamin-c. All the animals were served with normal saline (N/S 4mL/kg p.o), captopril (50mg/kg/day p.o), sodium hydrosulfide (NaHS) (56µmol/kg i.p), spironolactone (50mg/kg/day s.c), metformin (500mg/kg/day p.o) and vitamin-c (50mg/kg p.o) on daily basis for next 4 weeks, respectively. Metabolic studies, H2S levels, renal hemodynamics and oxidative stress markers were analyzed at 0, 14 and 28 days followed by histopathological analysis of renal tissues. The results showed decreased H2S levels, body weight, sodium to potassium ratio, glutathione (GSH), superoxide dismutase (SOD), total antioxidant assay (T-AOC) with malondialdehyde (MDA) and blood glucose levels significantly increased among diabetic rats. Treatment with captopril, NaHS, metformin, spironolactone and vitamin C showed significant improvement among renal hemodynamics and oxidative stress markers, respectively. But treatment groups like NaHS in combination with captopril and metformin showed more pronounced effects. The observations suggest that H2S mediated protective effects on STZ-induced diabetic nephropathy may be associated with reduced oxidative stress via augmenting the antioxidant effect.


Mitochondrial structural changes and dysfunction are associated with experimental allergic asthma.
An imbalance between Th1 and Th2 immune response is crucial for the development of pathophysiological features of asthma. A Th2-dominant response produces oxidative stress in the airways, and it is thought to be one of the crucial components of asthma pathogenesis. Although mitochondrion is a crucial organelle to produce endogenous reactive oxygen species, its involvement in this process remains unexplored as yet. We demonstrate in this study that OVA-induced experimental allergic asthma in BALB/c mice is associated with mitochondrial dysfunction, such as reduction of cytochrome c oxidase activity in lung mitochondria, reduction in the expression of subunit III of cytochrome c oxidase in bronchial epithelium, appearance of cytochrome c in the lung cytosol, decreased lung ATP levels, reduction in the expression of 17 kDa of complex I in bronchial epithelium, and mitochondrial ultrastructural changes such as loss of cristae and swelling. However, there was no change in the expression of subunits II and III of cytochrome c oxidase. Interestingly, administration of IL-4 mAb reversed these mitochondrial dysfunction and structural changes. In contrast, IFN-gamma mAb administration neither reversed nor further deteriorated the mitochondrial dysfunction and structural changes compared with control asthmatic mice administered with isotypic control Ab, although airway hyperresponsiveness deteriorated further. These results suggest that mitochondrial structural changes and dysfunction are associated with allergic asthma. These findings may help in the development of novel drug molecules targeting mitochondria for the treatment of asthma.


How does a 4-week motor-cognitive training affect choice reaction, dynamic balance and cognitive performance ability? A randomized controlled trial in well-trained, young, healthy participants.
We aimed to investigate the potential effects of a 4-week motor-cognitive dual-task training on cognitive and motor function as well as exercise motivation in young, healthy, and active adults. A total of 26 participants (age 25 ± 2 years; 10 women) were randomly allocated to either the intervention group or a control group. The intervention group performed a motor-cognitive training (3×/week), while the participants of the control group received no intervention. Before and after the intervention period of 4 weeks, all participants underwent cognitive (d2-test, Trail Making Test) and motor (lower-body choice reaction test and time to stabilization test) assessments. Following each of the 12 workouts, self-reported assessments (rating of perceived exertion, enjoyment and pleasant anticipation of the next training session) were done. Analyses of covariances and 95% confidence intervals plotting for between group and time effects were performed. Data from 24 participants were analysed. No pre- to post-intervention improvement nor a between-group difference regarding motor outcomes (choice-reaction: F = 0.5; time to stabilization test: F = 0.7; p > 0.05) occurred. No significant training-induced changes were found in the cognitive tests (D2: F = 0.02; Trail Making Test A: F = 0.24; Trail Making Test B: F = 0.002; p > 0.05). Both enjoyment and anticipation of the next workout were rated as high. The neuro-motor training appears to have no significant effects on motor and cognitive function in healthy, young and physically active adults. This might be explained in part by the participants' very high motor and cognitive abilities, the comparably low training intensity or the programme duration. The high degree of exercise enjoyment, however, may qualify the training as a facilitator to initiate and maintain regular physical activity. The moderate to vigorous intensity levels further point towards potential health-enhancing cardiorespiratory effects.


Rigorous criterion for characterizing correlated multiphoton emissions.
Strong correlation of photons, particularly in the single-photon regime, has recently been exploited for various applications in quantum information processing. Existing correlation measurements, however, do not fully characterize multi-photon correlation in a relevant context and may pose limitations in practical situations. We propose a conceptually rigorous, but easy-to-implement, criterion for detecting correlated multi-photon emission out of a quantum optical system, drawn from the context of wavefunction collapse. We illustrate the robustness of our approach against experimental limitations by considering an anharmonic optical system.


Erection of Ceratonova n. gen. (Myxosporea: Ceratomyxidae) to encompass freshwater species C. gasterostea n. sp. from threespine stickleback (Gasterosteus aculeatus) and C. shasta n. comb. from salmonid fishes.
Ceratonova gasterostea n. gen. n. sp. is described from the intestine of freshwater Gasterosteus aculeatus L. from the Klamath River, California. Myxospores are arcuate, 22.4 ± 2.6 μm thick, 5.2 ± 0.4 μm long, posterior angle 45° ± 24°, with 2 sub-spherical polar capsules, diameter 2.3 ± 0.2 μm, which lie adjacent to the suture. Its ribosomal small subunit sequence was most similar to an intestinal parasite of salmonid fishes, Ceratomyxa shasta (97%, 1,671/1,692 nucleotides), and distinct from all other Ceratomyxa species (<85%), which are typically coelozoic parasites in the gall bladder or urinary system of marine fishes. We propose erection of genus Ceratonova to contain both intestinal, freshwater species and reassign the salmonid parasite as Ceratonova shasta n. comb.


Soybean Golgi SNARE 12 protein interacts with Soybean mosaic virus encoded P3N-PIPO protein.
Soybean mosaic virus (SMV), a member of the Potyvirus genus, is one of the most prevalent and devastating viral pathogens in soybean-growing regions worldwide. It is generally accepted that symptom development of a viral plant disease results from molecular interactions between the virus and its host plant. P3N-PIPO, as a trans-frame protein consisting of the amino-terminal half of P3 fused to PIPO of the Potyvirus, plays a key role of viral cell-to-cell movement. This study provides evidence that Golgi SNARE 12 (designated as GOS12) protein of soybean interacts with SMV P3N-PIPO via a two-hybrid yeast system by screening a soybean cDNA library. Bimolecular fluorescence complementation (BiFC) assay further confirmed the interaction, which occurred in the cytomembrane of Nicotiana benthamiana cells. We also confirmed that the domain involved in the interaction is PIPO domain of P3N-PIPO by two-hybrid yeast system and BiFC. It is possible that the GOS12 is an essential host factor for PD targeting of P3N-PIPO protein of potyvirus.


Primary ciliary dyskinesia in the genomics age.
Primary ciliary dyskinesia is a genetically and clinically heterogeneous syndrome. Impaired function of motile cilia causes failure of mucociliary clearance. Patients typically present with neonatal respiratory distress of unknown cause and then continue to have a daily wet cough, recurrent chest infections, perennial rhinosinusitis, otitis media with effusion, and bronchiectasis. Approximately 50% of patients have situs inversus, and infertility is common. While understanding of the underlying genetics and disease mechanisms have substantially advanced in recent years, there remains a paucity of evidence for treatment. Next-generation sequencing has increased gene discovery, and mutations in more than 40 genes have been reported to cause primary ciliary dyskinesia, with many other genes likely to be discovered. Increased knowledge of cilia genes is challenging perceptions of the clinical phenotype, as some genes reported in the last 5 years are associated with mild respiratory disease. Developments in genomics and molecular medicine are rapidly improving diagnosis, and a genetic cause can be identified in approximately 70% of patients known to have primary ciliary dyskinesia. Groups are now investigating novel and personalised treatments, although gene therapies are unlikely to be available in the near future.


A systematic review of nonabsorbable, absorbable, and steroid-impregnated spacers following endoscopic sinus surgery.
Middle meatal (MM) spacers may reduce adhesions following endoscopic sinus surgery (ESS). However, there is no consensus as to which materials and adjuncts are the most effective for this purpose. The primary objective of this study was to examine the effectiveness of absorbable spacers (AS) vs nonabsorbable spacers (NAS) in reducing adhesions following ESS for chronic sinusitis. Secondarily, we assessed steroidal vs nonsteroidal MM spacers in reducing adhesions. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic review of randomized controlled trials (RCTs) was conducted and a meta-analysis on relevant outcome data was performed. Electronic search was done using OVID MEDLINE, EMBASE, Cochrane Central register of Controlled Trials, and Web of Science. Independent data extraction and evaluation was conducted. For our primary objective, 6 RCTs were included in the systematic review. A pooled estimate of relevant trials revealed a nonsignificant trend favoring AS in reducing adhesion formation compared to NAS (Relative Risk (RR), 0.40; 95% confidence interval [CI], 0.15-1.03). This trend was not apparent if NAS are left for greater than 48 hours after ESS. For our secondary objective, we identified 5 RCTs that compared steroidal vs nonsteroidal spacers. Although a pooled analysis could not be achieved due to inconsistent reporting of MM adhesions, 2 studies demonstrated significantly less adhesions in the steroidal spacer group. Comparison between NAS and AS showed that there was no significant difference in adhesion rates if NAS are used for at least 48 hours after surgery. Steroidal spacers may reduce adhesions, but more consistent data reporting is required for meta-analysis.


Recovery from opioid anesthesia: the clinical implication of context-sensitive half-times.
The context-sensitive half-time, the time required for a 50% decrease in drug concentration, has been proposed to predict the speed of recovery after infusions of i.v. anesthetics. We studied 40 patients to compare the clinical recovery characteristics of alfentanil and sufentanil. Patients were randomly allocated to receive either sufentanil/propofol (Group 1) or alfentanil/propofol (Group 2) total i.v. anesthesia by target-controlled infusions (TCI), assuming an equipotency ratio of 500:1. After discontinuation, times to tracheal extubation and to discharge from the postanesthesia care unit were measured, as were drug concentrations up to 24 h. The TCI bias was -17.1% for sufentanil and -16.9% for alfentanil. We found no difference in mean extubation times between the groups (48.7 min in Group 1 versus 46.4 min in Group 2), whereas discharge criteria were fulfilled significantly (P = 0.039) earlier after alfentanil (99.5 min) compared with sufentanil (131.3 min). The relative decrement values to tracheal extubation were 62.1% for sufentanil and 48.0% for alfentanil, compared with 75.7% and 65.0% for discharge, respectively. Based on a difference in propofol requirements, we suggest an actual sufentanil to alfentanil equipotency ratio of 1:300. We conclude that the decay in pharmacodynamic effect is not only the result of pharmacokinetics. Computer simulations may help to anticipate the clinical behavior of anesthetic drugs. In a clinical setting, we tested whether the recovery characteristics after i.v. anesthesia could be explained by a pharmaco-kinetic value, which describes the decline of drug concentrations in the body. This was not fully achieved.


[Importance of nutritional treatment in the inflammatory process of rheumatoid arthritis patients; a review].
Clinical characteristics of rheumatoid arthritis (AR) justify its multidisciplinary approach, within which nutritional intervention is included. Certain nutritional components influence the cellular metabolism and interfere in the pathological inflammatory process, so that they may act as coadjutant in the treatment of many inflammatory diseases, among which AR is in - cluded. To analyze the evidence related to the therapeutical effects of the polyunsaturated fatty acids, mediterranean diet, olive oil and certain antioxidant nutrients, on chronic inflammation and AR symptomatology. Non-Systematic revision in Cochrane, Pubmed, Scopus, Sportdiscus and Amed, from 2003 to March 2013, and subsequent applications of inclusion and exclusion criteria. Evidence on polyunsaturated fatty acids suggests that they produce clinical improvement and inhibitory effects over the AR inflammatory response. As regards the mediterranean diet, evidence points out that it decreases both pain and disease activity. In the case of the olive oil, although there are not enough studies, some effects are observed such as inflammatory markers reduction and oxidative stress inhibition. Finally, limited and contradictory evidence is found regarding the effectiveness of antioxidants. There exist studies which suggest that some dietetic elements (polyunsaturated fatty acids, mediterranean diet and antioxidants) have anti-inflammatory effects and decrease AR disease activity. More studies are required to empower the results on those aspects where evidence is still non conclusive.


Exploiting cross-priming to generate protective CD8 T-cell immunity rapidly.
The number of memory CD8 T cells generated by infection or vaccination correlates strongly with the degree of protection observed in infection and tumor models. Therefore, rapid induction of protective numbers of effector and memory CD8 T cells may be crucial in the case of malignancy, pandemic infection, or bioterrorism. Many studies have shown that amplifying T-cell numbers by prime-boost vaccination is most effective with a substantial time interval between immunizations. In contrast, immunization with peptide-coated mature dendritic cells (DCs) results in a CD8 T-cell response exhibiting accelerated acquisition of memory characteristics, including the ability to respond to booster immunization within days of initial priming. However, personalized DC immunization is too costly, labor intensive, and time-consuming for large-scale vaccination. Here, we demonstrate that in vivo cross-priming with cell-associated antigens or antigen-coated, biodegradable microspheres in the absence of adjuvant quickly generates CD8 T cells that display the phenotype and function of long-term memory populations. Importantly, cross-primed CD8 T cells can respond to booster immunization within days of the initial immunization to generate rapidly large numbers of effector and memory T cells that can protect against bacterial, viral, and parasitic infections, including lethal influenza and malaria-causing Plasmodium infection. Thus, accelerated CD8 T-cell memory after in vivo cross-priming in the absence of adjuvant is generalizable and can be exploited to generate protective immunity rapidly.


Pathological function of Ca2+-sensing receptor in pulmonary arterial hypertension.
Pulmonary arterial hypertension (PAH) is defined as an intractable disease characterized by a progressive elevation of pulmonary vascular resistance (PVR) and pulmonary arterial pressure (PAP), leading to right heart failure and premature death. The five-year survival rate after diagnosis is approximately 57%. Although extensive research has identified some factors associated with the cause of PAH, the etiology and pathogenesis remain unclear. In addition to Ca(2+) channel blockers (nifedipine, diltiazem), three categories of drug have been developed for the treatment of PAH based on the pathological mechanisms: prostacyclin and its analogues (epoprostenol, treprostinil, iloprost), endothelin receptor antagonists (bosentan, ambrisentan), and phosphodiesterase type 5 inhibitors (sildenafil, tadalafil). However, screening of novel types of drug acting on the signal pathway associated with the pathological mechanism underlying PAH is ongoing. We recently found that the extracellular Ca(2+)-sensing receptor (CaSR), which belongs to family C of the G protein-coupled receptor (GPCR) superfamily, is upregulated in pulmonary arterial smooth muscle cells (PASMCs) from patients with idiopathic PAH (IPAH). The upregulated CaSR is necessary for the enhanced Ca(2+) signaling and the augmented cell proliferation in PASMCs from IPAH patients. Most importantly, blockage of CaSR with an antagonist, NPS2143, prevents the development of pulmonary hypertension and right ventricular hypertrophy in animal models of pulmonary hypertension. The use of calcilytics, antagonists of CaSR, may be a novel therapeutic approach for PAH patients.


Leptin and the regulation of body weigh.
The cloning of the ob gene and its gene product, leptin, has led to the elucidation of a robust physiologic system that maintains fat stores at a relatively constant level. Leptin is a peptide hormone secreted by adipose tissue in proportion to its mass. Recessive mutations in the leptin gene are associated with massive obesity in mice and humans, establishing a genetic basis for obesity. Leptin circulates in blood and acts on the brain to regulate food intake and energy expenditure. When fat mass falls, plasma leptin levels fall, stimulating appetite and suppressing energy expenditure until fat mass is restored. When fat mass increases, leptin levels increase, suppressing appetite until weight is lost. This system maintains homeostatic control of adipose tissue mass. The discovery of leptin has advanced our understanding of metabolic disease in a number of respects. Its identification has revealed a new endocrine system regulating body weight. This system provides a means by which changes in nutritional state regulate other physiologic systems. A number of leptin deficiency syndromes that are treatable with leptin replacement have been identified. The majority of obese subjects are leptin resistant, which establishes that obesity is the result of hormone resistance. Leptin treatment results in weight loss in a subset of obese patients and can also synergize with other anti-obesity agents to reduce weight in the general population. Leptin provides an entry point for studying a complex human behavior. Finally, this research has established that there is a powerful biological basis for obesity, a fact that is (correctly) changing public perception about the pathogenesis of this medical condition.


Role of 11 beta-hydroxyprogesterone in biosynthesis of corticosterone from progesterone in guinea pig adrenals.
Slices of male guinea pig adrenals were incubated with trace amount of 3-H7 alpha-progesterone and the percentage of incorporation of 3-H into various steroids at various intervals was measured. It was found that 3-H accumulates predominately in 11 beta-hydroxyprogesterone as compared to DOC. This indicates that, in guinea pig adrenals, 11 beta-hydroxyprogesterone plays a dominant role in a formation of corticosterone from progesterone. The observed sequence of changes in progesterone label accumulation in 17-hydroxycompounds indicates they pathway of cortisol synthesis through the intermediate formation of 17-hydroxyprogesterone and 11-deoxycortisol.


Efficacy of the CDK7 Inhibitor on EMT-Associated Resistance to 3rd Generation EGFR-TKIs in Non-Small Cell Lung Cancer Cell Lines.
Epithelial to mesenchymal transition (EMT) is associated with resistance during EGFR tyrosine kinase inhibitor (EGFR-TKI) therapy. Here, we investigated whether EMT is associated with acquired resistance to 3rd generation EGFR-TKIs, and we explored the effects of cyclin-dependent kinase 7 (CDK7) inhibitors on EMT-mediated EGFR-TKIs resistance in non-small cell lung cancer (NSCLC). We established 3rd generation EGFR-TKI resistant cell lines (H1975/WR and H1975/OR) via repeated exposure to WZ4002 and osimertinib. The two resistant cell lines showed phenotypic changes to a spindle-cell shape, had a reduction of epithelial marker proteins, an induction of vimentin expression, and enhanced cellular mobility. The EMT-related resistant cells had higher sensitivity to THZ1 than the parental cells, although THZ1 treatment did not inhibit EGFR activity. This phenomenon was also observed in TGF-β1 induced EMT cell lines. THZ1 treatment induced G2/M cell cycle arrest and apoptosis in all of the cell lines. In addition, THZ1 treatment led to drug-tolerant, EMT-related resistant cells, and these THZ1-tolerant cells partially recovered their sensitivity to 3rd generation EGFR-TKIs. Taken together, EMT was associated with acquired resistance to 3rd generation EGFR-TKIs, and CDK7 inhibitors could potentially be used as a therapeutic strategy to overcome EMT associated EGFR-TKI resistance in NSCLC.


Effects of aldosterone and potassium-sparing diuretics on electrical potential differences across the distal nephron.
We have previously shown that the transtubular potential of the rabbit cortical collecting tubule varies in concert with changes in plasma mineralocorticoid levels, while the potential of the distal convoluted tubule is invariant with such changes. In the present studies we have examined the effects of in vitro addition of d-aldosterone to isolated tubules, as well as the effects of triamterene and spirolactone. d-Aldosterone (0.2 mum added to the perfusate or 1 muM added to the bathing medium) resulted in a marked stimulation of the transtubular potential difference (lumen-negative) after a short latent period. d-Aldosterone had no effect on the potential difference of distal convoluted tubules of intact or adrenalectomized rabbits. Both the magnitude of the response and the length of the latent period in the cortical collecting tubule after aldosterone were markedly temperature-dependent. Triamterene caused a gradual but reversible inhibition of the potential difference in the cortical collecting tubule but had no effect in the distal tubule. Spirolactone, when added before aldosterone, blocked the electrical response to the hormone in the cortical collecting tubule, and produced a gradual inhibition of the potential difference in mineralocorticoid-stimulated tubules. Spirolactone had no effect on the potential difference of the distal tubule. We conclude that (a) the influence of aldosterone on the potential across the distal nephron is restricted to the distal convoluted tubule, (b) the electrical response to aldosterone and the latent period are temperature-dependent, (c) the response to aldosterone is blocked by spirolactone, and (d) triamterene inhibits the potential difference only in the cortical collecting tubule.


Circadian rhythmicity in the activities of adenylate cyclase and phosphodiesterase in synchronously dividing and stationary-phase cultures of the achlorophyllous ZC mutant of Euglena gracilis.
Key factors in the adenosine 3',5'-cyclic monophosphate (cyclic AMP) metabolic pathway are two enzymes responsible for its generation and degradation, namely, adenylate cyclase (AC) and phosphodiesterase (PDE). In LD: 12,12 (12 h light, 12 h dark), these enzymes were found to undergo bimodal, circadian variation of activity in both dividing and nondividing cultures of the photosynthesis-deficient, achlorophyllous ZC mutant of Euglena gracilis Klebs (Z). Maximal AC activity occurred 2 h after the onset of the light interval (CT 02) and at the beginning of darkness (CT 12-14); these times corresponded to the acrophase profile for the rhythmic changes in cyclic AMP content that have been previously reported. The activity of PDE also exhibited a daily oscillation, but with an inverse phase pattern. Both the AC and PDE activity rhythms persisted after the cultures were transferred from LD: 12,12 to constant darkness. The activity of AC was activated significantly in vivo by forskolin at the trough phase (CT 20), while that of PDE was inhibited by 3-isobutyl-1-methyl-xanthine (IBMX) at its peak phase. These results indicate that the rhythms of both AC and PDE may be the main factors generating the circadian oscillations of cyclic AMP content in Euglena, which appear to be under control of an endogenous pacemaker.


The Importance of Energy Balance.
Globally, bodyweight and obesity are rising in both the developing and developed world. To maintain a stable bodyweight, energy intake must, over time, exactly equal energy expenditure, a state known as energy balance. An understanding of the physiologic control of energy balance may be useful for designing interventions to tackle the obesity epidemic worldwide. Obesity occurs when the body's energy balance is positive (i.e. when energy intake exceeds energy expenditure). Human physiology is biased towards maintaining energy balance at high levels of energy intake and expenditure. As a result, strategies to combat obesity should include a focus on increasing physical activity along with strategies for modifying food intake. An understanding of energy balance leads to the conclusion that prevention of weight gain should be easier than treatment of obesity. Components of energy balance are interdependent, and weight loss requires major behaviour changes, which trigger compensatory decreases in energy expenditure that facilitate weight regain. Prevention of weight gain can be accomplished by smaller behaviour changes. In addition to being easier to sustain than larger behaviour changes, smaller ones produce less compensation by the energy balance regulatory system. It has been estimated that relatively small changes in energy intake and expenditure totaling 100 kcal per day could arrest weight gain in most people. Interventions that advocate small changes have shown promising levels of success.


Automated Online-Sampling Multidimensional Liquid Chromatography with Feedback-Control Capability as a Framework for Real-Time Monitoring of mAb Critical Quality Attributes in Multiple Bioreactors.
Real-time monitoring of biopharmaceutical reactors is becoming increasingly important as the processes become more complex. During the continuous manufacturing of monoclonal antibodies (mAbs), the desired mAb product is continually created and collected over a 30 day process, where there can be changes in quality over that time. Liquid chromatography (LC) is the workhorse instrumentation capable of measuring mAb concentration as well as quality attributes such as aggregation, charge variants, oxidation, etc. However, traditional offline sampling is too infrequent to fully characterize bioprocesses, and the typical time from sample generation to data analysis and reporting can take weeks. To circumvent these limitations, an automated online sampling multidimensional workflow was developed to enable streamlined measurements of mAb concentration, aggregation, and charge variants. This analytical framework also facilitates automated data export for real-time analysis of up to six bioreactors, including feedback-controlling capability using readily available LC technology. This workflow increases the data points per bioreactor, improving the understanding of each experiment while also reducing the data turnaround time from weeks to hours. Examples of effective real-time analyses of mAb critical quality attributes are illustrated, showing substantial throughput improvements and accurate results while minimizing labor and manual intervention.