Expression and characterization of the nitrile reductase queF from E. coli

AbstractThe expression and characterization of a nitrile reductase from Escherichia coli K-12 (EcoNR), a newly discovered enzyme class, is described. This enzyme has a potential application for an alternative nitrile reduction pathway. The enzyme activity towards its natural substrate, preQ0, was demonstrated and optimal working conditions were found to be at 37°C and at pH 7 with Tris buffer

Differences in Mouse Hepatic Thyroid Hormone Transporter Expression with Age and Hyperthyroidism

Background: Clinical features of thyroid dysfunction vary with age, and an oligosymptomatic presentation of hyperthyroidism is frequently observed in the elderly. This suggests age modulation of thyroid hormone (TH) action, which may occur, for example, by alterations in TH production, metabolism and/or TH action in target organs. Objectives: In this paper, we address possible changes in TH transporter expression in liver tissues as a mechanism of age-dependent variation in TH action. Methods: Chronic hyperthyroidism was induced in 4- and 20-month-old C57BL6/NTac male mice (n = 8-10) by intraperitoneal injections of 1 µg/g body weight L-thyroxine (T4) every 48 h over 7 weeks. Control animals were injected with PBS. Total RNA was isolated from liver samples for analysis of the TH transporter and TH-responsive gene expression. TH concentrations were determined in mice sera. Results: Baseline serum free T4 (fT4) concentrations were significantly higher in euthyroid young compared to old mice. T4 treatment increased total T4, fT4 and free triiodothyronine to comparable concentrations in young and old mice. In the euthyroid state, TH transporter expression was significantly higher in old than in young mice, except for Mct8 and Oatp1a1 expression levels. Hyperthyroidism resulted in upregulation of Mct10, Lat1 and Lat2 in liver tissue, while Oatp1a1, Oatp1b2 and Oatp1a4 expression was downregulated. This effect was preserved in old animals. Conclusion: Here, we show age-dependent differences in TH transporter mRNA expression in the euthyroid and hyperthyroid state of mice focusing on the liver as a classical TH target organ

Sex-specific phenotypes of hyperthyroidism and hypothyroidism in mice

Background Thyroid dysfunction is more common in the female population, however, the impact of sex on disease characteristics has rarely been addressed. Using a murine model, we asked whether sex has an influence on phenotypes, thyroid hormone status, and thyroid hormone tissue response in hyper- and hypothyroidism. Methods Hypo- and hyperthyroidism were induced in 5 -month-old female and male wildtype C57BL/6N mice, by LoI/MMI/ClO4 − or T4 i.p. treatment over 7 weeks, and control animals underwent sham treatment (N = 8 animals/sex/treatment). Animals were investigated for impact of sex on body weight, food and water intake, body temperature, heart rate, behaviour (locomotor activity, motor coordination, and strength), liver function, serum thyroid hormone status, and cellular TH effects on gene expression in brown adipose tissue, heart, and liver. Results Male and female mice showed significant differences in behavioural, functional, metabolic, biochemical, and molecular traits of hyper- and hypothyroidism. Hyperthyroidism resulted in increased locomotor activity in female mice but decreased muscle strength and motor coordination preferably in male animals. Hypothyroidism led to increased water intake in male but not female mice and significantly higher serum cholesterol in male mice. Natural sex differences in body temperature, body weight gain, food and water intake were preserved under hyperthyroid conditions. In contrast, natural sex differences in heart rate disappeared with TH excess and deprivation. The variations of hyper- or hypothyroid traits of male and female mice were not explained by classical T3/T4 serum state. TH serum concentrations were significantly increased in female mice under hyperthyroidism, but no sex differences were found under eu- or hypothyroid conditions. Interestingly, analysis of expression of TH target genes and TH transporters revealed little sex dependency in heart, while sex differences in target genes were present in liver and brown adipose tissue in line with altered functional and metabolic traits of hyper- and hypothyroidism. Conclusions These data demonstrate that the phenotypes of hypo- and hyperthyroidism differ between male and female mice and indicate that sex is an important modifier of phenotypic manifestations

Image acquisition and planimetry systems to develop wounding techniques in 3D wound model

Wound healing represents a complex biological repair process. Established 2D monolayers and wounding techniques investigate cell migration, but do not represent coordinated multi-cellular systems. We aim to use wound surface area measurements obtained from image acquisition and planimetry systems to establish our wounding technique and in vitro organotypic tissue. These systems will be used in our future wound healing treatment studies to assess the rate of wound closure in response to wound healing treatment with light therapy (photobiomodulation). The image acquisition and planimetry systems were developed, calibrated, and verified to measure wound surface area in vitro. The system consists of a recording system (Sony DSC HX60, 20.4 M Pixel, 1/2.3″ CMOS sensor) and calibrated with 1mm scale paper. Macro photography with an optical zoom magnification of 2:1 achieves sufficient resolution to evaluate the 3mm wound size and healing growth. The camera system was leveled with an aluminum construction to ensure constant distance and orientation of the images. The JPG-format images were processed with a planimetry system in MATLAB. Edge detection enables definition of the wounded area. Wound area can be calculated with surface integrals. To separate the wounded area from the background, the image was filtered in several steps. Agar models, injured through several test persons with different levels of experience, were used as pilot data to test the planimetry software. These image acquisition and planimetry systems support the development of our wound healing research. The reproducibility of our wounding technique can be assessed by the variability in initial wound surface area. Also, wound healing treatment effects can be assessed by the change in rate of wound closure. These techniques represent the foundations of our wound model, wounding technique, and analysis systems in our ongoing studies in wound healing and therapy

The Senses of Agency and Ownership in Patients With Borderline Personality Disorder

Patients with borderline personality disorder (BPD) not only experience a strong instability in their affect and interpersonal relations but also disturbances in their self-experience, including dissociation and body-alienation symptoms. It is not yet understood whether an altered sense of ownership (SoO) or sense of agency (SoA) may contribute to these disturbances. One recent hypothesis is that patients with BPD have a reduced sense of self and are therefore more likely to misattribute external objects or actions to their own self than healthy individuals. The present study followed up this hypothesis by investigating whether BPD patients have a more flexible body representation than healthy participants. More specifically, the active rubber hand illusion (aRHI) was applied to 21 patients with BPD and the same number of healthy participants. Using established subjective, electrodermal, and behavioral measures, the participants' SoO and SoA were assessed during the aRHI. The findings show self-reported evidence for higher SoO under anatomical hand congruency as compared to anatomical incongruency, but no evidence for group differences between BPD patients and healthy participants. This finding is inconsistent with previous findings of an enhanced SoO-related body plasticity in BPD patients. Regarding SoA, the findings show self-report evidence of higher SoA in BPD patients versus healthy participants, although this group difference was not evident in the implicit SoA measure (intentional binding). In summary, the present study only reveals partial evidence for a higher body plasticity in BPD patients. Instead, the observed variability in results appears better explainable by some generally elevated perceptual suggestibility of BPD individuals

Sex-specific phenotypes of hyperthyroidism and hypothyroidism in aged mice

Abstract Background Sex and age play a role in the prevalence of thyroid dysfunction (TD), but their interrelationship for manifestation of hyper- and hypothyroidism is still not well understood. Using a murine model, we asked whether sex impacts the phenotypes of hyper- and hypothyroidism at two life stages. Methods Hyper- and hypothyroidism were induced by i.p. T4 or MMI/ClO4-/LoI treatment over 7 weeks in 12- and 20-months-old female and male C57BL/6N mice. Control animals underwent PBS treatment (n = 7–11 animals/sex/treatment). Animals were investigated for impact of sex on body weight, food and water intake, body temperature, heart rate, behaviour (locomotor activity, motor coordination and strength) and serum thyroid hormone (TH) status. Results Distinct sex impact was found in eu- and hyperthyroid mice, while phenotypic traits of hypothyroidism were similar in male and female mice. No sex difference was found in TH status of euthyroid mice; however, T4 treatment resulted in twofold higher TT4, FT4 and FT3 serum concentrations in adult and old females compared to male animals. Hyperthyroid females consistently showed higher locomotor activity and better coordination but more impairment of muscle function by TH excess at adult age. Importantly and in contrast to male mice, adult and old hyperthyroid female mice showed increased body weight. Higher body temperature in female mice was confirmed in all age groups. No sex impact was found on heart rate irrespective of TH status in adult and old mice. Conclusions By comparison of male and female mice with TD at two life stages, we found that sex modulates TH action in an organ- and function-specific manner. Sex differences were more pronounced under hyperthyroid conditions. Importantly, sex-specific differences in features of TD in adult and old mice were not conclusively explained by serum TH status in mice

The Plastic Surgery Workforce and Its Role in Low-income Countries

Background: Plastic surgery varies in scope, especially in different settings. This study aimed to quantify the plastic surgery workforce in low-income countries (LICs), understand commonly treated conditions by plastic surgeons working in these settings, and assess the impact on reducing global disease burden. Methods: We queried national and international surgery societies, plastic surgery societies, and non-governmental organizations to identify surgeons living and working in LICs who provide plastic surgical care using a cross-sectional survey. Respondents reported practice setting, training experience, income sources, and perceived barriers to care. Surgeons ranked commonly treated conditions and reported which of the Disease Control Priorities-3 essential surgery procedures they perform. Results: An estimated 63 surgeons who consider themselves plastic surgeons were identified from 15 LICs, with no surgeons identified in the remaining 16 LICs. Responses were obtained from 43 surgeons (70.5%). The 3 most commonly reported conditions treated were burns, trauma, and cleft deformities. Of the 44 "Essential Surgical Package'' procedures, 37 were performed by respondents, with the most common being skin graft (73% of surgeons performing), cleft lip/palate repair (66%), and amputations/escharotomy (61%). The most commonly cited barrier to care was insufficient equipment. Only 9% and 5% of surgeons believed that there are enough plastic surgeons to handle the burden in their local region and country, respectively. Conclusions: Plastic surgery plays a significant role in the coverage of essential surgical conditions in LICs. Continued expansion of the plastic surgical workforce and accompanying infrastructure is critical to meet unmet surgical burden in low- and middle-income countries

Chemical speciation of environmentally significant metals with inorganic ligands. Part 4: The Cd2+ + OH–, Cl–, CO32–, SO42–, and PO43– systems (IUPAC Technical Report)

The numerical modeling of Cd-II speciation amongst the environmental inorganic ligands Cl-, OH-, CO32-, SO42-, and PO43- requires reliable values for the relevant stability (formation) constants. This paper compiles and provides a critical review of these constants and related thermodynamic data. It recommends values of log(10) beta(p,q,r) valid at I-m = 0 mol kg(-1) and 25 degrees C (298.15 K), along with the equations and empirical reaction ion interaction coefficients, Delta epsilon, required to calculate log(10)beta(p),(,q,r) values at higher ionic strengths using the Bronsted-Guggenheim-Scatchard specific ion interaction theory (SIT). Values for the corresponding reaction enthalpies, Delta H-r, are reported where available. Unfortunately, with the exception of the Cd-II-chlorido system and (at low ionic strengths) the Cd-II-sulfato system, the equilibrium reactions for the title systems are relatively poorly characterized. In weakly acidic fresh water systems (-log(10){[H+]/c degrees} < 6), in the absence of organic ligands (e. g., humic substances), Cd-II speciation is dominated by Cd2+(aq), with CdSO4(aq) as a minor species. In this respect, Cd-II is similar to Cu-II [2007PBa] and Pb-II [2009PBa]. However, in weakly alkaline fresh water solutions, 7.5 < -log(10) {[H+]/c degrees} < 8.6, the speciation of Cd-II is still dominated by Cd2+(aq), whereas for Cu-II [2007PBa] and Pb-II [2009PBa] the carbonato-species MCO3(aq) dominates. In weakly acidic saline systems (-log(10) {[H+]/c degrees} < 6; -log(10) {[Cl-]/c degrees} < 2.0) the speciation is dominated by CdCln(2-n)+ complexes, (n = 1-3), with Cd2+(aq) as a minor species. This is qualitatively similar to the situation for Cu-II and Pb-II. However, in weakly alkaline saline solutions, including seawater, the chlorido-complexes still dominate the speciation of Cd-II because of the relatively low stability of CdCO3(aq). In contrast, the speciation of Cu-II [2007PBa] and Pb-II [2009PBa] in seawater is dominated by the respective species MCO3(aq). There is scope for additional high-quality measurements in the Cd2+ + H+ + CO32- system as the large uncertainties in the stability constants for the Cd2+-carbonato complexes significantly affect the speciation calculations

Additional file 1: Table S1-S3. of Sex-specific phenotypes of hyperthyroidism and hypothyroidism in aged mice

Statistical analysis of TT4, FT4, FT3 and TSH serum measurements in adult and old mice of both sexes. Two-way ANOVA followed by Bonferroni post hoc analysis was applied. Sex dependency was obvious as shown for Δ(mean female-mean male) values. Table S2 Statistical analysis of body temperature measurements in adult and old mice of both sexes. Two-way ANOVA followed by Bonferroni post hoc analysis was applied for hyper- and hypothyroid conditions. Average mean values of body temperature are shown as Δ(female-male). Table S3 Sex differences for area under curve (AUC) analysis of repeated body weight, food and water intake measurements. Two-way ANOVA followed by Bonferroni post hoc analysis was applied to AUC (±SEM) values, calculated by GraphPad Prism 7. (DOCX 18 kb