Objective Methotrexate (MTX) is the cornerstone in the treatment of rheumatoid arthritis (RA) patients. However, adherence to MTX therapy is not optimal, and instruments to assess medication non-adherence are warranted. To date there is no consensus on the best method to determine adherence to MTX. The aim of this study was to assess the correlation between adherence assessed with a Medication Event Monitoring System (MEMS) versus pill count, MTX-polyglutamate (PG) concentration and Compliance-Questionnaire-Rheumatology (CQR) in patients with established RA. Second, the correlations between these methods and the Disease Activity Scores of 28 joints (DAS28) were examined. Methods Adult RA patients currently treated with MTX were included. Multivariable linear and logistic regression were used, with adherence assessed with MEMS as dependent variable versus pill count, MTX-PG concentrations, CQR as independent variables, and DAS28 versus each of the four adherence measurements. Covariates were included, such as comedication, age and use of corticosteroids. Results 190 consecutive RA patients were included. Pill count was correlated with adherence assessed with MEMS (linear regression, β = .690, p = <.001), whereas CQR and MTX-PGs were not. Logistic regression confirmed the correlation between dichotomized adherence and pill count only (β = 5.64, p= <.001). No other correlations were found, neither for all adherence outcomes and DAS28. Conclusion Measuring adherence with MEMS is correlated with pill count, whereas other methods were not correlated with MEMS, nor with DAS28. Pill count can be used to estimate adherence to MTX therapy, in case MEMS is not achievable.

Mismatch negativity (MMN) and P3b are well known for their clinical utility. There exists no gold standard, however, to acquire these markers. This may explain why the within-individual sensitivity of MMN/P3b is often quite poor and why seemingly identical markers can behave differently across studies. Here we compare two traditional paradigms for MMN or P3b assessment with the recently more popular local-global paradigm which promises to assess MMN and P3b orthogonally within one oddball sequence. All three paradigms were administered to healthy participants (N=15) with concurrent EEG. A clear MMN and local effect were found for 15/15 participants. The P3b and global effect were found for 14/15 and 13/15 participants, respectively. There were no systematic differences between the global effect and P3b. Indeed, P3b amplitude was highly correlated across paradigms. The local effect differed clearly from the MMN, however. It occurred earlier compared to MMN and was followed by a much more prominent P3a effect. The two sets of peak latencies and amplitudes were also not correlated across paradigms. We conclude that the local-global paradigm is effective in evoking the traditional P3b component, but it does not capture the MMN. Caution should therefore be exercised when comparing the local effect and MMN across studies. Nevertheless, the within-individual sensitivity of both MMN and the local effect was satisfactory. The within-individual sensitivity of P3b was lower than expected in a healthy control group, which may explain the often-low sensitivity of P3b in patients with disorders of consciousness.

Traditional molecular biology tools have elucidated the identities and functions of RNA molecules, which are essential to the understanding of gene transcription and protein translation. Deepening this research field would further require the direct visualization of RNA dynamics such as the DNA-RNA interactions and RNA-protein interactions. Towards this goal, the rise of RNA imaging tools over the past 15 years has reformed how we looked at these processes. In this emerging topic, we first highlighted recent advances on three main RNA imaging tools based on the species of interacting molecules: RNA-RNA pairing, RNA-protein binding, and small molecule-RNA complex. We introduced the advantages of these tools from a technical viewpoint, including binding affinity, fluorescent turn-on ratio, stability, and impacts on targeted RNA. Next, we discussed new rising opportunities and future directions, echoing the state-of-the-art imaging tools in the fields of fluorescent proteins and small fluorescent molecules. Together, we believe this emerging field will bring new insights on how we study RNA biology in living systems.

This paper presents a framework combining Monte Carlo Simulation (MCS) and the Newmark sliding block model with Representative slip surfaces (RSS) (model II) and Multiple response surfaces method (MRSM) to conduct seismic reliability analysis and risk assessment of soil slopes. An empirical threshold is introduced to define the limit state function to identify the failure samples in MCS and the sliding area and Newmark sliding displacement are multiplied to quantify the failure consequence. The proposed methodology is illustrated through a soil slope with multiple layers. The calculation results demonstrate that traditional Newmark sliding block model (model I) tend to underestimate the variations of yield acceleration. Both the failure probability and landslide risk exhibit decreasing trends with the increase of threshold. Significant discrepancy in failure probability and landslide risk between two models is found even for a small threshold. It is therefore, the proposed methodology is highly recommended in seismic reliability analysis and risk assessment. The contributions of RSSs to the failure probability and landslide risk are insensitive to the variation of displacement thresholds.

Background and Purpose: 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1) catalyzes the oxoreduction of cortisone to cortisol, thereby amplifying levels of active glucocorticoids. It is considered a pharmaceutical target in metabolic disease and cognitive impairments. 11β-HSD1 also converts some 7oxo-steroids to their 7β-hydroxy forms. A recent study in mice described the ratio of tauroursodeoxycholic acid (TUDCA)/tauro-7oxolithocholic acid (T7oxoLCA) as a biomarker for decreased 11β-HSD1 oxoreductase activity. The present study aimed to evaluate the equivalent bile acid ratio glycoursodeoxycholic acid (GUDCA)/glyco-7oxolithocholic acid (G7oxoLCA) as a biomarker for pharmacological 11β-HSD1 inhibition in humans and compare it with the currently applied urinary (5α-tetrahydrocortisol+tetrahydrocortisol)/tetrahydrocortisone ((5αTHF+THF)/THE) ratio. Experimental Approach: Bile acid profiles were analyzed by ultra-HPLC tandem-MS in blood samples from two independent, double-blind placebo-controlled clinical studies on the orally administered selective 11β-HSD1 inhibitor AZD4017. The blood GUDCA/G7oxoLCA ratio was compared with the urinary tetrahydro-glucocorticoid ratio for the ability to detect 11β-HSD1 inhibition. Key Results: No significant alterations were observed in the bile acid profiles following 11β-HSD1 inhibition by AZD4017, except for an increase of the secondary bile acid G7oxoLCA. The enzyme product/substrate ratio GUDCA/G7oxoLCA was found to be more reliable to detect 11β-HSD1 inhibition than the absolute G7oxoLCA concentration in both cohorts. Comparison of the blood GUDCA/G7oxoLCA ratio with the urinary (5αTHF+THF)/THE ratio revealed that both ratios successfully detect 11β-HSD1 inhibition. Conclusion and Implications: 11β-HSD1 inhibition does not cause major alterations in bile acid homeostasis. The GUDCA/G7oxoLCA ratio represents the first blood biomarker of pharmacological 11β-HSD1 inhibition and may replace or complement the urinary (5αTHF+THF)/THE ratio biomarker.

Introduced alien species have direct and indirect effects on native communities, leading to lower taxonomic diversity and negative impacts on ecosystem functioning. Moreover, other aspects of diversity could be negatively affected, through alteration of functional and phylogenetic diversity of a community. This is particularly evident in habitats where human disturbance may favour alien species, posing an additional stressor on native communities. Following the community resistance hypothesis (higher diversity, higher resistance to invasion), we hypothesized: i) higher taxonomic, functional and phylogenetic diversity (TD, FD and PD respectively) in non-invaded bird communities (i.e. no alien bird species); and, ii) lower alien species impact on all diversity metrics in less human-disturbed areas. We surveyed bird communities in a modified Mediterranean landscape subject to varying levels of human disturbance. We tested whether TD, FD and PD indices were significantly different between non-invaded and invaded bird communities, and assessed the effect of landscape composition and configuration on these indices. We found that non-invaded communities retained higher TD and FD than invaded communities. Alien birds occupied novel parts of the functional space in invaded communities, but that they did not fully compensate for the taxonomic and functional diversity loss caused by the absence of native species. These results were consistent across different habitats, suggesting weak environmental filtering of communities. Generally, both communities were negatively affected by more human-disturbed areas (e.g. agriculture and urban areas) and enhanced by forest areas and by landscape heterogeneity. Our results suggest that the occurrence of alien birds negatively affects TD and FD (but not PD) of bird community assemblages, but that this impact is stronger in human-modified landscapes. Therefore, since the conservation of biodiversity in anthropogenic habitats is a worldwide challenge, researchers should prioritize efforts to assess the effects of alien species on communities inhabiting those habitats.

This article explores recursive algorithms for parameter identification issues of Hammerstein output-error systems. The proposed approach includes the key term separation auxiliary model recursive gradient algorithm, which utilizes the gradient search and the key term separation. To enhance computational efficiency, the system is decomposed into two or three subsystems through the hierarchical identification principle. Based on this, a key term separation auxiliary model two-stage recursive gradient algorithm and a key term separation auxiliary model three-stage recursive gradient algorithm are presented. The simulation results verify the validity of the obtained algorithms.

PD-1 and CTLA-4 can play an important role in addressing the issue of autoimmune diseases. PD-1 is a transmembrane glycoprotein expressed on T, B, and Dentric cells. This molecule functions as a checkpoint in T cell proliferation. Ligation of PD-1 with its ligands stimulates the production of IL-2, IL-7, IL-10, and IL-12 as well as other cytokines, which can inhibit cell proliferation and inflammation. Today, scientists attempt to protect against autoimmune diseases by PD-1 inhibitory signals. In this review, we discuss the structure, expression, and signaling pathway of PD-1. In addition, we discuss the importance of PD-1 in regulating several autoimmune diseases, reflecting how manipulating this molecule can be an effective method in the immunotherapy of some autoimmune diseases.

Background and objectives: A role of IL-10 is down-regulating T cell responses to schistosome antigens. Since SmATPDases can be correlated to modulation of the immune response, we evaluated the expression of enzymes in S. mansoni eggs. Patients/Methods: Fecal samples were collected from 40 infected individuals to detect coding regions of the SmATPDases. The cytokines was measured in supernatants of PBMC. The analysis was performed by the global median determination and set up high producers (HP) of cytokines. Results: Six individuals expressed SmATPDase1, six expressed SmATPDase2, and six expressed both enzymes. The group who expressed only SmATPDase1 showed a high frequency of IFN-γ, TNF, IL-4 HP, individuals who expressed only SmATPDase2 showed a high frequency of IFN-γ, IL-6, and IL-4 HP and individuals who expressed both enzymes showed a high frequency of IL-10 HP. In the group that showed expression both enzymes was observed lower indices the ratio between IFN-γ/IL-10. The positive correlation between infection intensity and IL-10 levels remained only in the positive SmATPDase group. The IL-10 is the only cytokine induced by the expression of both enzymes. Conclusions: The expression of both enzymes seems to be a factor that modulates the host immune response by inducing high IL-10 production.

Goat warble fly infestation (GWFI) is an economically important myiasis caused by larvae of Przhevalskiana silenus (Diptera, Oestridae), prevalent in countries of the Mediterranean Basin and Indian subcontinent. GWFI is characterized by the presence of subcutaneous warbles at the lumbar and sacral region of dorsum in the infested animal. The early larval instars (L1 and L2) remain inaccessible to physical detection due to their small size and subcutaneous presence thus causing prolonged economic loss to animal productivity. The early diagnostic intervention is needed during the disease monitoring and prophylactic management for effective control of the disease. The present study has developed an in-house dot-ELISA for the serodiagnosis of GWFI based on recombinant Hypodermin C (rHyC) antigen of Przhevalskiana silenus, expressed in E. coli. The purified protein was used for optimizing dot-ELISA in a checkerboard titration using goat warble fly infested serum as known positive. The optimized conditions require 188 ng of protein/dot, 1:800 dilution of serum sample, 1:4000 dilution of anti-goat IgG conjugate and 5% skim milk powder in phosphate buffer saline as blocking buffer. The assay was found to have a diagnostic sensitivity and specificity of 97.3% and 95.8%, respectively. The inter-rater reliability of dot ELISA with rHyC indirect ELISA was found to be almost perfect with a Cohen’s kappa index of 0.973. Further testing at ambient temperature (18 oC) and shorter incubation steps (30 min) supported suitability of the assay for field diagnosis of GWFI. The rHyC protein based Dot-ELISA was evaluated using random field serum samples suspected for GWFI. The present study provides the first report of a sensitive and specific dot-ELISA for early diagnosis of GWFI which is rapid and cost effective. The test may provide an effective tool for sustainable control of GWFI.

Insect venom allergy is the most frequent cause of anaphylaxis in Europe and possibly worldwide. The majority of systemic allergic reactions after insect stings are caused by Hymenoptera and among these, vespid genera induce most of the systemic sting reactions (SSR). Honey bees are the second leading cause of SSR. Depending on the global region, other Hymenoptera such as different ant genera are responsible for SSR. Widely distributed hornets and bumblebees or local vespid or bee genera rarely induce SSR. Hematophagous insects such as mosquitoes and horse flies usually cause (large) local reactions while SSR occasionally occur. This position paper aims to identify either rare or locally important insects causing SSR as well as rarely occurring SSR after stings or bites of widely distributed insects. We summarized relevant venom or saliva allergens and intended to identify possible cross-reactivities between the insect allergens. Moreover, we aimed to locate diagnostic tests for research and routine diagnosis, which are sometimes only regionally available. Finally, we gathered information on disposable immunotherapies. Major allergens of most insects were identified, and cross-reactivity between insects was frequently observed. While some diagnostics and immunotherapies are locally available, standardized skin tests and immunotherapies are generally lacking in rare insect allergy.

Microtiter plates are suitable for screening and process development of most microorganisms. They are currently the container of choice for high-throughput and small-scale microbial culture, but require optimization for specific work. This research presents a novel type of microtiter plate was developed using computational fluid dynamics (CFD) technology. The new plate provides high oxygen supply and optimal mixing effects for the fermentation culture of docosahexaenoic acid (DHA) producing strains, surpassing the conventional method of strains screening with shake flasks, which is insufficient. the shape of the microtiter plate was modified, and baffles were introduced to improve mass transfer and oxygen supply effects in the vibrating bioreactor. CFD technology was used to model the new plate’s characteristics, establishing the superiority of hexagonal microtiter plates with six baffles. Parameters in the incubation process, such as vibration frequency and liquid load, were optimized, and the final result achieved a KLa of 0.61s-1 and a volume power input of 2364 w/m3, which was 4-5 times better than the original 96-well plate. The culture results optimized by the model were also verified. Therefore, this new microtiter plate provides a powerful tool for future high-throughput screening of strains.

A document by Asma Albtoosh. Click on the document to view its contents.

We present a 10-year follow-up and describe our experience in managing a case of neonatal severe primary hyperparathyroidism (NSHPT) for the first time in Iran. Microcephaly, mental retardation and epilepsy may be long time sequels of NSHPT and the brain MRI findings are compatible with old hypoxic-ischemic event.

Tropical freshwater lakes are well-known for their high biodiversity, and the East African Great Lakes in particular are renowned for their endemic cichlid fish adaptive radiations. While comparative phylogenetic analyses of extant species flocks have revealed patterns and processes of their diversification, evolutionary trajectories within lineages, impacts of environmental drivers, or the scope and nature of now-extinct diversity remain largely unknown. Time-structured paleodata from geologically young fossil records, such as fossil counts and particularly ancient DNA data, would help fill this large knowledge gap. High ambient temperatures can be detrimental to the preservation of DNA, but refined methodology now allows data generation even from very poorly preserved samples. Here, we show for the first time that fish fossils from tropical lake sediments yield endogenous ancient DNA (aDNA). Despite generally low endogenous content and high sample drop-out, high-throughput sequencing and in some cases sequence capture allowed for taxonomic assignment to family or tribe level and phylogenetic placement of individuals. Even skeletal remains weighing less than 1 mg and up to 2700 years of age could be phylogenetically placed. We find that the relationship of degradation of aDNA with the thermal age of samples is similar to that described for terrestrial samples from cold environments adjusted for elevated temperatures. Success rates and aDNA preservation differed between the investigated lakes Chala, Kivu and Victoria, possibly caused by differences in water oxygenation at deposition. Our study demonstrates that sediments of tropical lakes preserve genetic information on rapidly diversifying taxa over time scales of millennia.

A document by Juan Hincapie-Castillo. Click on the document to view its contents.

An underutilized experimental design was used to isolate adapted mutants of the model bacterium Pseudomonas putida KT2440. The experimental design consisted of subjecting a random pool of mini-Tn5 mutants of P. putida KT2440 to several rounds of selection in the rhizosphere of soybean irrigated with NaCl solution. Isolated adapted mutants (MutAd) showed a mutation in a gene encoding the membrane-binding protein LapA, which is involved in the early stages of biofilm formation on abiotic surfaces. Two MutAd bacteria (MutAd160 and MutAd185) and a lapA deletion mutant were tested to study the effect of this gene on salt tolerance, rhizosphere fitness, extracellular polymeric substances (EPS) production, and plant growth promotion. The inability of the mutants to form biofilm did not hinder attachment to soybean seeds and roots. MutAd bacteria showed an overproduction of EPS when grown under saline conditions, which would compensate for the lack of biofilm formation. MutAd185 bacteria showed increased root attachment and growth promotion of soybean in slightly saline soils. The proposed experimental design would be useful to accelerate bacterial adaptation to the rhizosphere of plants under a given environmental condition, identify genetic mutations that benefit bacterial fitness in that condition, and thus increase their ability to promote plant growth.

Objective: To assess National Health Insurance Fund (NHIF) members’ level of understanding, experiences, and factors influencing their choice of NHIF-contracted outpatient facilities in Kenya. Methods: We conducted a cross-sectional qualitative study with NHIF members in two purposefully selected counties (Nyeri and Makueni counties) in Kenya. We collected data through 15 focus group discussions with NHIF members. Data were analysed using a framework analysis approach. Results: Urban-based NHIF members had a good understanding of the NHIF-contracted outpatient facility selection process and the approaches for choosing and changing providers, unlike their rural counterparts. While NHIF members were required to choose a provider before accessing care, the number of available alternative facilities was perceived to be inadequate. Finally, NHIF members identified seven factors they considered important when choosing an NHIF-contracted outpatient provider. Of these factors, the availability of drugs, distance from the household to the facility and waiting time at the facility until consultation were considered the most important. Conclusion: There is a need for the NHIF to prioritise awareness-raising approaches tailored to rural settings. Further, there is a need for the NHIF to contract more providers to both spur competition among providers and provide alternatives for members to choose from. Besides, NHIF members revealed the important factors they consider when selecting outpatient facilities. Consequently, NHIF should leverage the preferred factors when contracting healthcare providers. Similarly, healthcare providers should enhance the availability of drugs, reduce waiting times whilst improving their staff’s attitudes which would enhance user satisfaction and the quality of care provided.