Neuroleptic drug-induced coagulopathy: mechanism of reaction and duration of effect.
Patients (N = 32) with documented phenothiazine-induced reductions in platelet levels were evaluated after drug substitution with haloperidol; 28 (88%) patients had normal platelet levels after 1 year of treatment with haloperidol. Coagulation studies indicated that activated partial thromboplastin time (APTT) elevations were more common in patients previously treated with chlorpromazine than in those previously treated with fluphenazine. There was no relationship between attenuation of reduced platelet levels and reduced APTT ratios. This preliminary study indicates that there may be two mechanisms of phenothiazine-induced coagulopathy.


Investigation of structure and dynamics of the hydrated metal-organic framework MIL-53(Cr) using first-principles molecular dynamics.
The hydration behavior of metal-organic frameworks (MOFs) is of interest both from a practical and from a fundamental point of view: it is linked, on the one hand, to the hydrothermal stability (or instability) of the nanoporous material, which might limit its use in technological applications. On the other hand, it sheds light on the behavior of water in a strongly confined environment. Here, we use first-principles molecular dynamics (MD) to investigate two hydrated phases of the flexible MOF MIL-53(Cr), which adopts a narrow- or a large-pore form, depending on the water loading. Structure and dynamics of the two phases are thoroughly analyzed and compared, with a focus on the hydroxyl group of MIL-53(Cr) and the water molecules in the nanopores. Furthermore, the behavior of the confined water is compared to that of bulk water. Whereas in the narrow-pore form, water is adsorbed at specific crystalline sites, it shows a more disordered, bulk-like structure in the large-pore form. However, reorientation dynamics of water molecules in the latter is considerably slowed down with respect to bulk water, which highlights the confinement effect of the nanoporous framework.


Significance of urinary C3 excretion in glomerulonephritis.
The third component of complement (C3) was measured in the urine of 98 patients with a variety of renal diseases. Renal biopsy was performed on 83 of the patients and examined by light, electron, and immunofluorescence microscopy. Urinary C3 was detected in cases of membranous glomerulonephritis, mesangiocapillary glomerulonephritis, rapidly progressive glomerulonephritis, and renal amuloidosis. It was not detected in minimal lesion glomerulonephritis; in cases of proliferative glomerulonephritis it was detected only in those showing histological evidence of a progressive lesion. Concentrations were low or undetectable in cases of non-immunological renal diseases. There was a good correlation between urinary C3 concentrations and the deposition of C3 in glomerular capillary walls, as seen by immunofluorescence microscopy, and there was no correlation with the degree or selectivity of proteinuria. Urinary C3 excretion appears to be an accurate indicator of continuing activity of disease. It is suggested that the presence of C3 in urine is due to complement fixation by immune complexes in glomerular capillary walls, and that urinary C3 estimations have potential applications in the study of glomerulonephritis.


Chemical visualization of individual chondrocytes in articular cartilage by attenuated-total-reflection Fourier Transform Infrared Microimaging.
Fourier transform infrared imaging (FTIRI) and the attenuated total reflection Fourier transform infrared microimaging (ATR-FTIRM) were used to study the chemical and structural distributions of cellular components surrounding individual chondrocytes in canine humeral cartilage, at 6.25µm pixel resolution in FTIRI and 1.56µm pixel resolution in ATR-FTIRM. The chemical and structural distributions of the cellular components in chondrocytes and tissue can be successfully imaged in high resolution ATR-FTIRM. One can also study the territorial matrix of fine collagen fibrils surrounding the individual chondrocytes by the polarization experiments using the absorption ratio of amide I to amide II bands.


Magnetic resonance histology of the adult zebrafish brain: optimization of fixation and gadolinium contrast enhancement.
Magnetic resonance histology (MRH) has become a widespread tool to examine brain morphology in situ or ex vivo. Samples are routinely fixed and stained to allow for longer scan times with increased contrast and resolution. Although the zebrafish is an important model for neuroscience, to date most MRH studies have focused almost exclusively on mice. In this paper, we examined, for the first time, the zebrafish brain using MRH. We compared a range of fixatives, contrast agents, and fixation/staining durations to determine optimal imaging of the zebrafish brain. By quantifying the T(1), T(2), and T(2)* relaxation values, we demonstrated that ethanol and potassium permanganate are unviable for imaging and significant differences exist between mono and di-aldehydes. Furthermore, we compared two commercially available gadolinium-based contrast agents, Magnevist® and Optimark®, at five different concentrations. For both contrast agents, a concentration of 0.5% was determined to be ideal as it significantly shortened the T(1) but maintained a relatively long T(2) and T(2)*. Subsequently, we analyzed the duration of fixation/staining and established a period of 12 h, which best minimized T(1) values but maintained T(2) and T(2)* values. Finally, using this optimized fixation and staining protocol, we performed a gradient-echo T(2)*-weighted imaging to obtain an image set of the adult zebrafish brain at an isotropic resolution of 10 µm.


Risk factors in puerperal infection.
Puerperal infection is one of the most common obstetric complications which leads to certain serious sequelas for those parturients. The study of 92 postpartum women at Siriraj Hospital from April 1, 1980 to March 1, 1983 was conducted to investigate the risk factors of puerperal infection. All 92 cases were diagnosed for puerperal infection according to the criteria of the Joint Committee on Maternal Welfare. Irregular antenatal care or no antenatal care, sexual intercourse during the last week before delivery and pelvic examination during pregnancy were found to be important predisposing factors in the antepartum period. Pelvic examination during labor accounted for 78 per cent of the patients and played an important role, while operative obstetrics (30%) and concomitant diseases during pregnancy (27%) were the next significant factors.


Habit strength is predicted by activity dynamics in goal-directed brain systems during training.
Previous neuroscientific research revealed insights into the brain networks supporting goal-directed and habitual behavior, respectively. However, it remains unclear how these contribute to inter-individual differences in habit strength which is relevant for understanding not only normal behavior but also more severe dysregulations between these types of action control, such as in addiction. In the present fMRI study, we trained subjects on approach and avoidance behavior for an extended period of time before testing the habit strength of the acquired stimulus-response associations. We found that stronger habits were associated with a stronger decrease in inferior parietal lobule activity for approach and avoidance behavior and weaker vmPFC activity at the end of training for avoidance behavior, areas associated with the anticipation of outcome identity and value. VmPFC in particular showed markedly different activity dynamics during the training of approach and avoidance behavior. Furthermore, while ongoing training was accompanied by increasing functional connectivity between posterior putamen and premotor cortex, consistent with previous assumptions about the neural basis of increasing habitualization, this was not predictive of later habit strength. Together, our findings suggest that inter-individual differences in habitual behavior are driven by differences in the persistent involvement of brain areas supporting goal-directed behavior during training.


Nanomembrane Canister Architectures for the Visualization and Filtration of Oxyanion Toxins with One-Step Processing.
Nanomembrane canister-like architectures were fabricated by using hexagonal mesocylinder-shaped aluminosilica nanotubes (MNTs)-porous anodic alumina (PAA) hybrid nanochannels. The engineering pattern of the MNTs inside a 60 μm-long membrane channel enabled the creation of unique canister-like channel necks and cavities. The open-tubular canister architecture design provides controllable, reproducible, and one-step processing patterns of visual detection and rejection/permeation of oxyanion toxins such as selenite (SeO3(2-)) in aquatic environments (i.e., in ground and river water sources) in the Ibaraki Prefecture of Japan. The decoration of organic ligand moieties such as omega chrome black blue (OCG) into inorganic Al2O3@tubular SiO2/Al2O3 canister membrane channel cavities led to the fabrication of an optical nanomembrane sensor (ONS). The OCG ligand was not leached from the canister as observed in washing, sensing, and recovery assays of selenite anions in solution, which enabled its multiple reuse. The ONS makes a variety of alternate processing analyses of selective quantification, visual detection, rejection/permeation, and recovery of toxic selenite quick and simple without using complex instrumentation. Under optimal conditions, the ONS canister exhibited a high selectivity toward selenite anions relative to other ions and a low-level detection limit of 0.0093 μM. Real analytical data showed that approximately 96% of SeO3(2-) anions can be recovered from aquatic and wastewater samples. The ONS canister holds potential for field recovery applications of toxic selenite anions from water.


A high risk twin study of combat-related PTSD comorbidity.
Combat-related posttraumatic stress disorder (PTSD) is highly comorbid with other mental disorders. However, the nature of the relationship between PTSD and other mental disorders remains unclear. A discordant high-risk twin design was used on data from a sub-sample of the male-male twin pair members of the Vietnam Era Twin Registry to examine whether patterns of comorbidity are consistent with a psychopathological response to combat exposure or reflect familial vulnerability to psychopathology. Mental disorders were assessed via the Mental Health Diagnostic Interview Schedule Version III - Revised. Discordant monozygotic within-pair comparisons revealed that PTSD probands had higher symptom counts and diagnostic prevalences of mood and anxiety disorders than their non-combat exposed co-twins. Monozygotic co-twins of PTSD probands had significantly more mood disorder symptoms than monozygotic co-twins of combat controls or dizygotic co-twins of veterans with PTSD. These findings suggest that a) major depression, generalized anxiety disorder and panic disorder are part of a post-combat response syndrome; b) a shared familial vulnerability also contributes to the association between PTSD and major depression, PTSD and dysthymia, and c) this shared vulnerability is mediated by genetic factors.


Managing to harvest? Perspectives on the potential of aquaculture.
Aquaculture has been one of the most rapid and technically innovative of food production sectors globally, with significant investment, scientific and technical development and production growth in many parts of the world over the past two decades. While this has had a significant effect on the global supply of aquatic food products and had an important impact in rural and urban food supply and employment in many developing economies, growth and increasing internationalization has not been without concern for natural resource use, environmental impact and social disruption. The expectations for production and diversification are now significant and while the scientific and technical means are already available to meet much of the intended targets, practical constraints of investment, profitability, resource access and system efficiency are likely to become far more important constraints for the future. This review offers a contemporary perspective on the ways in which the sector might develop, its interactions with constraints and the strategies that may be required to ensure that future development is both positive and sustainable.


The IUPHAR/BPS Guide to PHARMACOLOGY in 2024.
The IUPHAR/BPS Guide to PHARMACOLOGY (GtoPdb; https://www.guidetopharmacology.org) is an open-access, expert-curated, online database that provides succinct overviews and key references for pharmacological targets and their recommended experimental ligands. It includes over 3039 protein targets and 12 163 ligand molecules, including approved drugs, small molecules, peptides and antibodies. Here, we report recent developments to the resource and describe expansion in content over the six database releases made during the last two years. The database update section of this paper focuses on two areas relating to important global health challenges. The first, SARS-CoV-2 COVID-19, remains a major concern and we describe our efforts to expand the database to include a new family of coronavirus proteins. The second area is antimicrobial resistance, for which we have extended our coverage of antibacterials in partnership with AntibioticDB, a collaboration that has continued through support from GARDP. We discuss other areas of curation and also focus on our external links to resources such as PubChem that bring important synergies to the resources.


STAMP2 increases oxidative stress and is critical for prostate cancer.
The six transmembrane protein of prostate 2 (STAMP2) is an androgen-regulated gene whose mRNA expression is increased in prostate cancer (PCa). Here, we show that STAMP2 protein expression is increased in human PCa compared with benign prostate that is also correlated with tumor grade and treatment response. We also show that STAMP2 significantly increased reactive oxygen species (ROS) in PCa cells through its iron reductase activity which also depleted NADPH levels. Knockdown of STAMP2 expression in PCa cells inhibited proliferation, colony formation, and anchorage-independent growth, and significantly increased apoptosis. Furthermore, STAMP2 effects were, at least in part, mediated by activating transcription factor 4 (ATF4), whose expression is regulated by ROS. Consistent with in vitro findings, silencing STAMP2 significantly inhibited PCa xenograft growth in mice. Finally, therapeutic silencing of STAMP2 by systemically administered nanoliposomal siRNA profoundly inhibited tumor growth in two established preclinical PCa models in mice. These data suggest that STAMP2 is required for PCa progression and thus may serve as a novel therapeutic target.


Discovery and characterization of prolactin neutralizing monoclonal antibodies for the treatment of female-prevalent pain disorders.
Prolactin (PRL) has recently been demonstrated to elicit female-selective nociceptor sensitization and increase pain-like behaviors in female animals. Here we report the discovery and characterization of first-in-class, humanized PRL neutralizing monoclonal antibodies (PRL mAbs). We obtained two potent and selective PRL mAbs, PL 200,031 and PL 200,039. PL 200,031 was engineered as human IgG1 whereas PL 200,039 was reformatted as human IgG4. Both mAbs have sub-nanomolar affinity for human PRL (hPRL) and produce concentration-dependent and complete inhibition of hPRL signaling at the hPRL receptor (hPRLR). These two PRL mAbs are selective for hPRL as they do not inhibit other hPRLR agonists such as human growth hormone or placental lactogen. They also cross-react with non-human primate PRL but not with rodent PRL. Further, both mAbs show long clearance half-lives after intravenous administration in FcRn-humanized mice. Consistent with their isotypes, these mAbs only differ in binding affinities to Fcγ receptors, as expected by design. Finally, PL 200,019, the murine parental mAb of PL 200,031 and PL 200,039, fully blocked stress-induced and PRL-dependent pain behaviors in female PRL-humanized mice, thereby providing in vivo preclinical proof-of-efficacy for PRL mAbs in mechanisms relevant to pain in females.


Decrease of signal intensity of myometrium and cervical stroma after ultrasmall superparamagnetic iron oxide (USPIO) particles administration: an MR finding with potential benefits in T staging of uterine neoplasms.
Following the empiric observation of a significant decrease of signal intensity of both myometrium and cervical stroma on ultrasmall superparamagnetic iron oxide (USPIO)-enhanced images, the aim of our study was to evaluate whether USPIO-enhanced T2*-weighted gradient echo (GRE) images might provide any potential advantage on T-staging of uterine malignancies having surgery and histology as standard of reference Seventeen female patients with known uterine malignancies underwent magnetic resonance (MR) imaging before and 24 hours after the intravenous administration of the USPIO agent. Imaging protocol included proton density-weighted turbo spin echo and T2*-weighted GRE sequences. Each patient underwent surgery within 14 days from the first MR examination, and histologic confirmation of tumor T-stage was obtained. Quantitative (calculation of signal-to-noise and contrast-to-noise ratios) and qualitative (visual assessment of T staging) analyses were performed on unenhanced and USPIO-enhanced images. Quantitative analysis showed a significantly lower (P < 0.05) signal-to-noise ratio of myometrium and cervical stroma on USPIO-enhanced compared with unenhanced images. In 15 of 17 patients (88.2%), the contrast-to-noise ratio between tumor and myometrium and between tumor and cervical stroma was higher on USPIO-enhanced compared with unenhanced images (P < 0.001). Qualitative analysis demonstrated that the GRE T2* USPIO-enhanced MR offers a better definition of the depth of tumor infiltration rather than the unenhanced GRE T2* images. The decrease of signal intensity of myometrium and cervical stroma on T2*-weighted GRE images after the intravenous administration of USPIO should be considered a constant and physiologic finding that improves tumor conspicuity in the majority of the cases, allowing more accurate T-staging of neoplastic lesions.


Pediatric Audiology Report: Assessment and Revision of an Audiology Report Written to Parents of Children With Hearing Impairment.
The purpose of this study was twofold: first, to evaluate a typical pediatric diagnostic audiology report to establish its readability and comprehensibility for parents and, second, to revise the report to improve its readability, as well as the comprehension, sense of self-efficacy, and positive opinions of parent readers. In Experiment 1, a mock audiology report was evaluated via a readability analysis and semistructured interviews with 5 parents. In Experiment 2, the report was revised using best practice guidelines and parental recommendations from Experiment 1. The revision was verified by randomly assigning 32 new parent participants to read either the revised or unrevised report before their comprehension, self-efficacy, and opinions were assessed. In Experiment 1, results confirmed that the report was difficult to read and understand. In Experiment 2, parents who read the revised report had significantly greater comprehension, self-efficacy, and opinion ratings than those who read the unrevised report. In addition, the readability of the revised report was markedly improved compared with the unrevised report. This study shows that pediatric diagnostic audiology reports can be revised to adhere to best practice guidelines and yield improved readability, in addition to improving the comprehension, sense of self-efficacy, and positive opinions of parents of children with hearing impairment.


Hyperkalemia revisited.
Hyperkalemia is a common clinical condition that can induce deadly cardiac arrhythmias. Electrocardiographic manifestations of hyperkalemia vary from the classic sine-wave rhythm, which occurs in severe hyperkalemia, to nonspecific repolarization abnormalities seen with mild elevations of serum potassium. We present a case of hyperkalemia, initially diagnosed as ventricular tachycardia, to demonstrate how difficult hyperkalemia can be to diagnose. An in-depth review of hyperkalemia is presented, examining the electrophysiologic and electrocardiographic changes that occur as serum potassium levels increase. The treatment for hyperkalemia is then discussed, with an emphasis on the mechanisms by which each intervention lowers serum potassium levels. An extensive literature review has been performed to present a comprehensive review of the causes and treatment of hyperkalemia.


Characterization of wound microbes in epidermolysis bullosa: Results from the epidermolysis bullosa clinical characterization and outcomes database.
Patients with epidermolysis bullosa (EB) require care of wounds that are colonized or infected with bacteria. A subset of EB patients are at risk for squamous cell carcinoma, and bacterial-host interactions have been considered in this risk. The EB Clinical Characterization and Outcomes Database serves as a repository of information from EB patients at multiple centers in the United States and Canada. Access to this resource enabled broad-scale analysis of wound cultures. A retrospective analysis of 739 wound cultures from 158 patients from 13 centers between 2001 and 2018. Of 152 patients with a positive culture, Staphylococcus aureus (SA) was recovered from 131 patients (86%), Pseudomonas aeruginosa (PA) from 56 (37%), and Streptococcus pyogenes (GAS) from 34 (22%). Sixty-eight percent of patients had cultures positive for methicillin-sensitive SA, and 47%, methicillin-resistant SA (18 patients had cultures that grew both methicillin-susceptible and methicillin-resistant SA at different points in time). Of 15 patients with SA-positive cultures with recorded mupirocin susceptibility testing, 11 had mupirocin-susceptible SA and 6 patients mupirocin-resistant SA (2 patients grew both mupirocin-susceptible and mupirocin-resistant SA). SCC was reported in 23 patients in the entire database, of whom 10 had documented wound cultures positive for SA, PA, and Proteus species in 90%, 50%, and 20% of cases, respectively. SA and PA were the most commonly isolated bacteria from wounds. Methicillin resistance and mupirocin resistance were reported in 47% and 40% of patients tested, respectively, highlighting the importance of ongoing antimicrobial strategies to limit antibiotic resistance.


Targeting HER2 heterogeneity in breast and gastrointestinal cancers.
About 20% of breast and gastric cancers and 3% of colorectal carcinomas overexpress the human epidermal growth factor receptor 2 (HER2) and are sensitive to HER2-directed agents. The expression of HER2 may differ within the same tumoral lesion (spatial intralesional heterogeneity), from different tumor locations (spatial interlesional heterogeneity), and throughout treatments (temporal heterogeneity). Spatial and temporal heterogeneity may impact on response and resistance to HER2-targeting agents and its prevalence and predictive role changes across HER2-overexpressing solid tumors. Therefore, the definition and the characterization of HER2 heterogeneity pose many challenges and its implementation as a reproducible predictive biomarker would help in guiding treatment modulation.


Introducing an environmental microbiome to axenic Aedes aegypti mosquitoes documents bacterial responses to a blood meal.
The blood meal of the female mosquito serves as a nutrition source to support egg development, so is an important aspect of its biology. Yet, the roles the microbiome may play in blood digestion are poorly characterized. We employed axenic mosquitoes to investigate how the microbiome differs between mosquitoes reared in the insectary versus mosquitoes that acquire their microbiome from the environment. Environmental microbiomes were more diverse and showed larger temporal shifts over the course of blood digestion. Importantly, only bacteria from the environmental microbiome performed hemolysis in culture, pointing to functional differences between bacterial populations. These data highlight that taxonomic differences between the microbiomes of insectary-reared and wild mosquitoes are potentially also related to their functional ecology. Thus, axenic mosquitoes colonized with environmental bacteria offer a way to investigate the role of bacteria from the wild in mosquito processes such as blood digestion, under controlled laboratory conditions.


Differential anti-ischaemic effects of muscarinic receptor blockade in patients with obstructive coronary artery disease; impaired vs normal left ventricular function.
In patients with coronary artery disease acetylcholine (a muscarinic agonist) causes vasoconstriction. The effect of atropine (a muscarinic antagonist) on coronary vasotone in patients with normal or impaired left ventricular function is unknown. Twenty-four patients who required atropine infusion (to supplement heart rate response) during atrial pacing (pacing was conducted to assess ischaemia as part of an experimental protocol) were studied; 17 patients had normal and seven impaired left ventricular function (ejection fraction < or =0.40). Two control groups were selected from a large database (from patients in whom atrial pacing was carried out but to whom atropine was not administered) to match the normal (n=20) and dysfunction (n=10) groups. In the normal left ventricular function group atropine increased rate pressure product by 12 +/- 4%, as compared to those without atropine (P < 0.05). Left ventricular end diastolic pressure increased less in the atropine group (+40 +/- 8% vs +78 +/- 6%;P < 0.05). Arterial norepinephrine increased similarly in both groups, but coronary flow (as assessed by using a thermodiluting method in the coronary sinus) increased 23 +/ -4% more in the atropine group (P < 0.05). Further, there were lower levels of myocardial lactate production and ST-segment depression in the atropine group [lactate extraction +13 +/- 6% (atropine) vs -19 +/- 4% (controls), ST-segment depression 1. 3 +/- 0.6 (atropine) vs 1.8 +/- 0.2 mm (control), both P < 0.05 between groups]. In contrast, in the dysfunction group the overall effect of atropine was less pronounced. In patients with normal left ventricular function atropine improves coronary flow and reduces myocardial lactate production and ST-segment depression during atrial pacing, suggesting a reduction in myocardial ischaemia.


Mating induces pupillary dilatation in female rats: role of pelvic nerve.
In the present study, we demonstrate that natural or artificial vaginal stimulation exerts a powerful effect on autonomic activity, as measured by pupillary diameter. Pupillary diameter was recorded approximately 3-5 sec after copulatory mounts, intromissions, ejaculations or experimenter delivered stimulation. The pupil was magnified 43X and its diameter was measured directly from a calibrated video screen. Mating tests were performed on 13 female rats which received at least one complete ejaculatory series. Compared to the premount (baseline) value (1.01 mm), mean pupillary diameters were significantly greater during (a) mounts without intromissions (1.20 mm; 20% increase), (b) mounts with intromissions (1.34 mm; 33% increase), or (c) ejaculation (1.49 mm; 48% increase). Mean pupillary diameter during the post-ejaculatory interval (0.95 mm; 6% decrease) did not differ significantly from the premount baseline. Since the mean pupillary diameter at the last intromission before ejaculation was significantly larger (by 18%) than the diameter at the first intromission, repeated intromissions may generate a gradually increasing net sympathetic response reaching a peak at ejaculation. In order to identify the adequate stimuli for increasing pupillary diameter, we applied external genital stimulation (flank-perineal palpation), internal genital stimulation (probing the vaginal cervix with a glass rod), or both combined. Vaginal stimulation alone produced near-maximal pupillary dilatation; external genital stimulation was less effective, and combined internal and external genital stimulation was more effective. Thus, vaginal afferent activity is an adequate stimulus to produce pupillary dilatation. Furthermore, bilateral pelvic neurectomy markedly attenuated the effect of vaginal stimulation on dilating the pupil. Since pupillary diameter is at least partly a function of autonomic tone, this "bioassay" indicates that genital stimulation may produce a net sympathetic activating effect.


Cryobacterium mesophilum sp. nov., a novel mesophilic bacterium.
A novel mesophilic, Gram-positive, cream- to pale yellow-pigmented, aerobic bacterium, designated MSL-15T, was isolated from a soil sample collected from Bigeum Island, South Korea. The cells were catalase-positive, motile and irregular rods. The diamino acid content of the cell-wall peptidoglycan was determined to be 2,4-diaminobutyric acid. Strain MSL-15T grew well between 20 and 28 degrees C, with optimum growth occurring at 25-28 degrees C. The major fatty acids were anteiso-C15 : 0, iso-C16 : 0, anteiso-C17 : 0 and iso-C15 : 0. The genomic DNA G+C content was 70.0 mol%. A phylogenetic analysis based on 16S rRNA gene sequences indicated that strain MSL-15T was associated with the genus Cryobacterium and exhibited 96.6 % sequence similarity to Cryobacterium psychrotolerans JCM 13925T and 94.8 % similarity to Cryobacterium psychrophilum JCM 1463T. Phenotypic, chemotaxonomic and phylogenetic data revealed that strain MSL-15T represents a novel species within the genus Cryobacterium, for which the name Cryobacterium mesophilum is proposed. The type strain is MSL-15T (=DSM 19267T=KCTC 19270T).


[Effect of intensive operator activity on lipid peroxidation processes in the human body].
It has been demonstrated that the stress associated with an active mental work for 2 hours leads to a high rate of lipid peroxidation which results in a greater amount of products of lipid peroxidation in blood and pentane in exhaled air. Simultaneous measurements of blood catecholamines have shown their significant increase immediately after exposure to the stress effect. It can therefore be concluded that a stress situation enhances lipid peroxidation in the human body.


Preventing unintended proteolysis in plant protein biofactories.
Numerous reports have been published over the last decade assessing the potential of plants as useful hosts for the heterologous expression of clinically useful proteins. Significant progress has been made, in particular, in optimizing transgene transcription and translation in plants, and in elucidating the complex post-translational modifications of proteins typical of the plant cell machinery. In this article, we address the important issue of recombinant protein degradation in plant expression platforms, which directly impacts on the final yield, homogeneity and overall quality of the resulting protein product. Unlike several more stable and structurally less complex pharmaceuticals, recombinant proteins present a natural tendency to structural heterogeneity, resulting in part from the inherent instability of polypeptide chains expressed in heterologous environments. Proteolytic processing, notably, may dramatically alter the structural integrity and overall accumulation of recombinant proteins in plant expression systems, both in planta during expression and ex planta after extraction. In this article, we describe the current strategies proposed to minimize protein hydrolysis in plant protein factories, including organ-specific transgene expression, organelle-specific protein targeting, the grafting of stabilizing protein domains to labile proteins, protein secretion in natural fluids and the co-expression of companion protease inhibitors.


Inverse ESPVR Estimation with Singularity Avoidance via Constrained EDPVR Parameter Optimization.
Left ventricular end-systolic elastance Ees, as an index of cardiac contractility, can play a key role in continuous patient monitoring during cardiac treatment scenarios such as drug therapies. The clinical feasibility of Ees estimation remains challenging because most techniques have been built on left ventricular pressure and volume, which are difficult to measure or estimate in the regular ICU/CCU setting. The purpose of this paper is to propose and validate a novel approach to estimate Ees, which is independent of left ventricular pressure and volume. Our methods first derive an analytical representation of Ees as the inverse function of the gradient of the Frank-Starling Curve based on cardiac mechanics. Second, elucidating the mechanism of singularities in the inverse function, we derive multiple conditions in both end-systolic pressure-volume relationship (ESPVR) and end-diastolic pressure-volume relationship (EDPVR) parameters to avoid these singularities analytically. Third, we formulate a constrained nonlinear least squares problem to optimize both ESPVR and EDPVR parameters simultaneously to avoid singularities. The effectiveness of the proposed method in avoiding singularities was evaluated in an animal experiment. Compared to the conventional Ees estimation by linear regression, our proposed method reproduced in-vivo hemodynamics more accurately when simulating the estimated Ees variation during drug administration. Our method can be applied using the available data in the regular ICU/CCU setting. The improved clinical feasibility can support not only physicians' decision-making, including adjusting drug dosages in current clinical treatment, but also a closed-loop hemodynamic control system requiring accurate continuous Ees estimation.


Modifying Kohlrausch's Law to Describe Nonaqueous Electrolytes for Lithium-Ion Batteries.
To develop next-generation lithium-ion batteries with enhanced stability and safety, it is crucial to understand the physicochemical principles of nonaqueous electrolytes. Kohlrausch's law describes a linear decrease in the molar conductivity (Λ) with respect to the square root of the molarity of strong electrolytes at lower concentrations. This empirical law explains the impeded ionic mobility at higher concentrations due to ionic interactions, i.e., relaxation and asymmetric effects. However, this law does not hold at higher concentrations due to the ionic association that alleviates the ionic interactions and retards the decrease in the Λ. Especially, the anomalously stagnant decrease in the Λ near the solubility limit has not been clearly explained, calling for the consideration of other concentration-dependent factors such as the mean activity coefficient (γ±), viscosity (η), and dielectric constant (ε). Herein, we develop a systematic method to modify Kohlrausch's law. First, we install the ionic association constant, and the theoretical estimation is compared with the experimental results to induce the correction function that is related with the previously neglected concentration-dependent factors. Thus, the induced correction function was close to the rectified linear unit (ReLU) function, which has been widely used in the field of artificial intelligence. The modified Kohlrausch's law with the ReLU-type correction function provides a highly precise and reliable data fitting, and the fitted parameters showed clear concentration dependency and straightforward interpretability. As a result, this method effectively generalized Kohlrausch's law for nonaqueous electrolytes at higher concentrations up to the solubility limit of 3.0-3.5 M. Moreover, the modified Kohlrausch's law inspired us to discover the physical origins of the anomalously stagnant Λ profiles near the solubility limit; and the most relevant physical origin of the anomaly was the concentration dependency of the γ± and η, which grow exponentially above a critical concentration.


Motivation to learn: an overview of contemporary theories.
To succinctly summarise five contemporary theories about motivation to learn, articulate key intersections and distinctions among these theories, and identify important considerations for future research. Motivation has been defined as the process whereby goal-directed activities are initiated and sustained. In expectancy-value theory, motivation is a function of the expectation of success and perceived value. Attribution theory focuses on the causal attributions learners create to explain the results of an activity, and classifies these in terms of their locus, stability and controllability. Social- cognitive theory emphasises self-efficacy as the primary driver of motivated action, and also identifies cues that influence future self-efficacy and support self-regulated learning. Goal orientation theory suggests that learners tend to engage in tasks with concerns about mastering the content (mastery goal, arising from a 'growth' mindset regarding intelligence and learning) or about doing better than others or avoiding failure (performance goals, arising from a 'fixed' mindset). Finally, self-determination theory proposes that optimal performance results from actions motivated by intrinsic interests or by extrinsic values that have become integrated and internalised. Satisfying basic psychosocial needs of autonomy, competence and relatedness promotes such motivation. Looking across all five theories, we note recurrent themes of competence, value, attributions, and interactions between individuals and the learning context. To avoid conceptual confusion, and perhaps more importantly to maximise the theory-building potential of their work, researchers must be careful (and precise) in how they define, operationalise and measure different motivational constructs. We suggest that motivation research continue to build theory and extend it to health professions domains, identify key outcomes and outcome measures, and test practical educational applications of the principles thus derived.


A voltage sensor-domain protein is a voltage-gated proton channel.
Voltage-gated proton channels have been widely observed but have not been identified at a molecular level. Here we report that a four-transmembrane protein similar to the voltage-sensor domain of voltage-gated ion channels is a voltage-gated proton channel. Cells overexpressing this protein showed depolarization-induced outward currents accompanied by tail currents. Current reversal occured at equilibrium potentials for protons. The currents exhibited pH-dependent gating and zinc ion sensitivity, two features which are characteristic of voltage-gated proton channels. Responses of voltage dependence to sequence changes suggest that mouse voltage-sensor domain-only protein is itself a channel, rather than a regulator of another channel protein.


Immunoprofiling of Cell Wall Carbohydrate Modifications During Flooding-Induced Aerenchyma Formation in Fabaceae Roots.
Understanding plant adaptation mechanisms to prolonged water immersion provides options for genetic modification of existing crops to create cultivars more tolerant of periodic flooding. An important advancement in understanding flooding adaptation would be to elucidate mechanisms, such as aerenchyma air-space formation induced by hypoxic conditions, consistent with prolonged immersion. Lysigenous aerenchyma formation occurs through programmed cell death (PCD), which may entail the chemical modification of polysaccharides in root tissue cell walls. We investigated if a relationship exists between modification of pectic polysaccharides through de-methyl esterification (DME) and the formation of root aerenchyma in select Fabaceae species. To test this hypothesis, we first characterized the progression of aerenchyma formation within the vascular stele of three different legumes-Pisum sativum, Cicer arietinum, and Phaseolus coccineus-through traditional light microscopy histological staining and scanning electron microscopy. We assessed alterations in stele morphology, cavity dimensions, and cell wall chemistry. Then we conducted an immunolabeling protocol to detect specific degrees of DME among species during a 48-hour flooding time series. Additionally, we performed an enzymatic pretreatment to remove select cell wall polymers prior to immunolabeling for DME pectins. We were able to determine that all species possessed similar aerenchyma formation mechanisms that begin with degradation of root vascular stele metaxylem cells. Immunolabeling results demonstrated DME occurs prior to aerenchyma formation and prepares vascular tissues for the beginning of cavity formation in flooded roots. Furthermore, enzymatic pretreatment demonstrated that removal of cellulose and select hemicellulosic carbohydrates unmasks additional antigen binding sites for DME pectin antibodies. These results suggest that additional carbohydrate modification may be required to permit DME and subsequent enzyme activity to form aerenchyma. By providing a greater understanding of cell wall pectin remodeling among legume species, we encourage further investigation into the mechanism of carbohydrate modifications during aerenchyma formation and possible avenues for flood-tolerance improvement of legume crops.


Sigma receptors and their ligands in cancer biology: overview and new perspectives for cancer therapy.
A large number of drugs are known to bind with high affinity to sigma receptors (sigma-Rs) and have been used in the clinic to treat mental disorders for many years. However, recent publications highlighting sigma-R overexpression in many cancer tissues suggest potential applications for sigma-R ligands in cancer diagnosis and therapy. The present review focuses on the involvement of sigma-Rs in cancer biology and the potential therapeutic contributions of their pharmacologic ligands in oncology. After summarizing the current and general knowledge regarding sigma-Rs, we detail data reported in the particular context of oncology. We then investigate the potential and specific signal transduction pathways and mechanisms involved in the actions of sigma-R ligands in cancer biology. These processes include modulations of (1) the plasma membrane and lipid raft components, (2) intracellular calcium levels, (3) cytoskeletal protein functions, and (4) endoplasmic reticulum stress. Finally, we conclude by speculating on the roles of sigma-R overexpression and sigma-R ligands in cancer biology and offer perspectives on cancer therapy.


Analysis of isoniazid-resistant transposon mutants of Mycobacterium smegmatis.
The emergence of multidrug-resistant tuberculosis has renewed interest in the study of drug resistance in mycobacteria with the objective of improved chemotherapy. The genetic basis of isoniazid resistance in a model mycobacterium was studied. Eleven isoniazid-resistant mutants of Mycobacterium smegmatis were created using transposon mutagenesis. Genetic and enzymatic characterisation of the mutants showed that katG, encoding T-catalase, was inactivated. The nucleotide sequence of M. smegmatis katG was determined and the mutation sites mapped demonstrating that both the amino and carboxyl halves of T-catalase are important for enzymatic activity.


Predictors of mortality of patients with acute respiratory failure secondary to chronic obstructive pulmonary disease admitted to an intensive care unit: a one year study.
Patients with acute exacerbation of chronic obstructive pulmonary disease (COPD) commonly require hospitalization and admission to intensive care unit (ICU). It is useful to identify patients at the time of admission who are likely to have poor outcome. This study was carried out to define the predictors of mortality in patients with acute exacerbation of COPD and to device a scoring system using the baseline physiological variables for prognosticating these patients. Eighty-two patients with acute respiratory failure secondary to COPD admitted to medical ICU over a one-year period were included. Clinical and demographic profile at the time of admission to ICU including APACHE II score and Glasgow coma scale were recorded at the time of admission to ICU. In addition, acid base disorders, renal functions, liver functions and serum albumin, were recorded at the time of presentation. Primary outcome measure was hospital mortality. Invasive ventilation was required in 69 patients (84.1%). Fifty-two patients survived to hospital discharge (63.4%). APACHE II score at the time of admission to ICU {odds ratio (95 % CI): 1.32 (1.138-1.532); p < 0.001} and serum albumin (done within 24 hours of admission) {odds ratio (95 % CI): 0.114 (0.03-0.432); p = 0.001}. An equation, constructed using the adjusted odds ratio for the two parameters, had an area under the ROC curve of 91.3%. For the choice of cut-off, sensitivity, specificity, positive and negative predictive value for predicting outcome was 90%, 86.5%, 79.4% and 93.7%. APACHE II score at admission and SA levels with in 24 hrs after admission are independent predictors of mortality for patients with COPD admitted to ICU. The equation derived from these two parameters is useful for predicting outcome of these patients.


The benefit of metformin in the treatment of pediatric non-alcoholic fatty liver disease: a systematic review and meta-analysis of randomized controlled trials.
This is the first meta-analysis of the available literature about the efficacy of metformin exclusively in pediatric patients with non-alcoholic fatty liver disease (NAFLD). We conducted a systematic literature search through major electronic databases till March 12, 2023, investigating the efficacy and safety of metformin in pediatric NAFLD. Weighted mean difference (WD) and standard deviation (SD) were used for continuous outcomes. In total, 4 randomized controlled trials (RCTs) with 309 pediatric patients with NAFLD were included in the meta-analysis. Metformin could not reach a statistically significant improvement in alanine aminotransferase (ALT) levels [(ALT: WMD = - 1.55 IU/L, 95% CI: - 5.38 to 2.28, I2 = 16%, p = 0.43), but had a statistically significant impact (p < 0.05) in insulin and HOMA-IR regulation, triglycerides, and high-density lipoprotein level improvement. Conclusion: According to the data of this meta-analysis, treatment with metformin failed to statistically improve liver enzymes but may be beneficial in the improvement of lipid parameters and insulin metabolism regulation in pediatric patients with NAFLD. As there are not enough available studies in the literature, the influence of metformin on liver ultrasonography or histology in pediatric NAFLD should be further analyzed in future studies. What is Known: • Lifestyle modification with weight loss through physical activity and dietary modification is the recommended treatment option for pediatric NAFLD. • Metformin may reduce steatosis on ultrasound and may have a beneficial role in liver histology collated with insulin resistance improvement. What is New: • Metformin may improve insulin sensitivity and lipid parameters in children with obesity and NAFLD. • Metformin does not have a significant effect on transaminase levels in children with obesity and NAFLD.


Hydroxylapatite growth on single-crystal rutile substrates.
Titanium is widely used as an implant material. In addition to the bulk properties of titanium, the biological response is to a large degree controlled via the surface. The native amorphous titanium oxide that forms spontaneously on the surface gives a very good biological response. Lately it has been shown that crystalline titanium oxides (rutile and anatase) have in vitro bioactive properties. In addition to its potential for new materials development, this finding also opens up for the possibility of studying the mechanisms of bioactivity on materials with strictly controlled surfaces. In this paper the mechanisms behind the in vitro bioactivity are studied, using rutile single crystals. Three single-crystal rutile substrates: (100), (110), and (001), and a polycrystalline rutile substrate obtained by physical vapour deposition were soaked in a phosphate buffered saline solution for up to 4 weeks. The hydroxylapatite films that formed were analysed by X-ray diffraction, scanning electron microscopy, focused ion beam, and transmission electron microscopy. The hydroxylapatite grew faster on the (001) surface than on the other two. It was also found that on the (001) surface the direction of fast growth in hydroxylapatite was aligned parallel to the surface. This is in contrast to the (110) rutile surface where the fast growth of the hydroxylapatite crystal was directed outwards from the surface. The (100) face had poor adhesion at the interface. The orientations of the precipitated crystallites play a significant role in the faster coverage of the (001) rutile face. Based on the experimental results, a model for the hydroxylapatite growth process is given.


Nanomedicine strategies for molecular targets with MRI and optical imaging.
The science of 'theranostics' plays a crucial role in personalized medicine, which represents the future of patient management. Over the last decade an increasing research effort has focused on the development of nanoparticle-based molecular-imaging and drug-delivery approaches, emerging as a multidisciplinary field that shows promise in understanding the components, processes, dynamics and therapies of a disease at a molecular level. The potential of nanometer-sized agents for early detection, diagnosis and personalized treatment of diseases is extraordinary. They have found applications in almost all clinically relevant biomedical imaging modality. In this review, a number of these approaches will be presented with a particular emphasis on MRI and optical imaging-based techniques. We have discussed both established molecular-imaging approaches and recently developed innovative strategies, highlighting the seminal studies and a number of successful examples of theranostic nanomedicine, especially in the areas of cardiovascular and cancer therapy.


Voices and challenges of marginalized and vulnerable groups in urban informal settlements in Nairobi, Kenya: building on a spectrum of community-based participatory research approaches.
Urbanization is rapidly increasing across Africa, including in Nairobi, Kenya. Many people, recent migrants and long-term residents, live within dense and dynamic urban informal settlements. These contexts are fluid and heterogeneous, and deepening the understanding of how vulnerabilities and marginalization are experienced is important to inform pointed action, service delivery and policy priorities. The aim of this paper is to explore vulnerabilities and marginalization within Korogocho and Viwandani informal settlements in Nairobi and generate lessons on the value of a spectrum of community based participatory research approaches for understanding health and well-being needs and pinpointing appropriate interventions. In the exploratory stages of our ARISE consortium research, we worked with co-researchers to use the following methods: social mapping, governance diaries, and photo voice. Social mapping (including the use of Focus Group Discussions) identified key vulnerable groups: marginalized and precarious child heads of households (CHHs), Persons with disability who face multiple discrimination and health challenges, and often isolated older adults; and their priority needs, including health, education, water and sanitation. The governance diaries generated an understanding of the perceptions of the particularly vulnerable and marginalized informal settlement residents regarding the various people and institutions with the power to influence health and wellbeing; while photo voice highlighted the lived experiences of vulnerability and marginality. Understanding and responding to fluid and intersecting marginalities and vulnerabilities within growing urban informal settlements is particularly critical to achieving inclusive urbanization, where no one is left behind, a theme central to the Sustainable Development Goals and Kenya's Vision 2030.


Steroid for epidural injection in spinal stenosis: a systematic review and meta-analysis.
To investigate the effectiveness and safety of epidural steroid injections in patients with lumbar spinal stenosis (LSS). We performed a search on the CENTRAL, Pubmed, Embase and Cochrane databases up to September 2014. We recovered 17 original articles, of which only 10 were in full compliance with the randomized controlled trial (RCT) criteria. These articles were reviewed in an independent and blinded way by two reviewers who were previously trained to extract data and score their quality by the criteria of the Cochrane Handbook (5.1.0). We accepted ten studies with 1,010 participants. There is minimal evidence that shows that epidural steroid injections are better than lidocaine alone, regardless of the mode of epidural injection. There is a fair short-term and long-term benefit for treating spinal stenosis with local anesthetic and steroids. This meta-analysis suggests that epidural steroid injections provide limited improvement in short-term and long-term benefits in LSS patients.


Carbon dioxide laser therapy.
A carbon dioxide laser coupled with a ventilating bronchoscope has been used to treat 135 patients with symptomatic obstruction of the trachea and main stem bronchi. This laser has the advantages of being controllable and hemostatic, and can be used repetitively. It does not interfere with other modes of treatment such as radiation therapy or chemotherapy.


Physical activity and fitness in the community: the Framingham Heart Study.
While greater physical activity (PA) is associated with improved health outcomes, the direct links between distinct components of PA, their changes over time, and cardiorespiratory fitness are incompletely understood. Maximum effort cardiopulmonary exercise testing (CPET) and objective PA measures [sedentary time (SED), steps/day, and moderate-vigorous PA (MVPA)] via accelerometers worn for 1 week concurrent with CPET and 7.8 years prior were obtained in 2070 Framingham Heart Study participants [age 54 ± 9 years, 51% women, SED 810 ± 83 min/day, steps/day 7737 ± 3520, MVPA 22.3 ± 20.3 min/day, peak oxygen uptake (VO2) 23.6 ± 6.9 mL/kg/min]. Adjusted for clinical risk factors, increases in steps/day and MVPA and reduced SED between the two assessments were associated with distinct aspects of cardiorespiratory fitness (measured by VO2) during initiation, early-moderate level, peak exercise, and recovery, with the highest effect estimates for MVPA (false discovery rate <5% for all). Findings were largely consistent across categories of age, sex, obesity, and cardiovascular risk. Increases of 17 min of MVPA/day [95% confidence interval (CI) 14-21] or 4312 steps/day (95% CI 3439-5781; ≈54 min at 80 steps/min), or reductions of 249 min of SED per day (95% CI 149-777) between the two exam cycles corresponded to a 5% (1.2 mL/kg/min) higher peak VO2. Individuals with high (above-mean) steps or MVPA demonstrated above average peak VO2 values regardless of whether they had high or low SED. Our findings provide a detailed assessment of relations of different types of PA with multidimensional cardiorespiratory fitness measures and suggest favourable longitudinal changes in PA (and MVPA in particular) are associated with greater objective fitness.


Ginger: a systematic review of clinical trials and recent advances in encapsulation of its bioactive compounds.
Recently, the numbers of studies on natural products have considerably increased owing to their exceptional biological activities and health benefits. Their pharmacological attributes have played an immense role in detecting natural and safe alternative therapeutics, consequently extending their industrial applications. In this line, ginger (Zingiber officinale) has been gaining wide attention owing to its bioactive compounds, such as phenolic and terpene compounds. Ginger has a great pharmacological and biological potential in the prevention and treatment of various diseases, namely colds, nausea, arthritis, migraines and hypertension. However, these bioactive compounds are unstable and susceptible to degradation, volatilization and oxidation during extraction and processing, mainly owing to their exposure to environments with adverse conditions, such as high temperature, the presence of O2 and light. In this sense, this current review covers a wide range of topics, starting from the chemical profile and biological properties of ginger bioactive compounds (GBCs), their clinical effectiveness for the treatment of diseases and the application of different encapsulation methods (molecular inclusion, spray drying, complex coacervation, ionic strength and nanoemulsions) to protect and improve their application in food products. This work summarizes the fundamental principles of, recent progress in and effectiveness of different methods regarding the physicochemical, structural and functional properties of encapsulated GBCs. The potential use of encapsulated GBCs as a promising active ingredient to be applied in different food products is discussed in detail.


Neurological assessments after treatment with the antimalarial β-arteether in neonatal and adult rats.
The World Health Organization currently recommends combinatorial treatment including artemisinins as first-line therapy against drug-resistant Plasmodium falciparum malaria. Although highly efficacious, artemisinin and its derivatives, including β-arteether (βAE), are associated with ototoxicity, tremors, and other autonomic and motor impairments in the clinic. Similar neurological symptoms, as well as brainstem lesions, have been observed in adult laboratory species (mice, rats, dogs, and non human primates) following acute treatment with βAE; however, few long-term, nonclinical studies have been conducted. Furthermore, the majority of deaths attributed to malarial infection occur in children under age five, yet no laboratory studies have been initiated in neonatal or juvenile animals. In the current study, neonatal 7-day-old rats were administered intramuscular doses of 1-90 mg/kg βAE in sesame oil for up to eight treatment cycles (one cycle=7 days treatment+7 days without treatment). Neonates were tested for changes in sensorimotor function, and the same animals were tested as adults in the Functional Observational Battery, for motor activity, and in the 8-arm radial maze. Pups receiving a single cycle of 60 or 90 mg/kg died within a week of treatment but had few behavioral changes and no brainstem pathology. In the long-term study, behavioral and motor changes and brainstem lesions were observed in a dose- and time-related manner. Rats given repeated cycles of 1 or 5mg/kg βAE showed subtle motor abnormalities (e.g., slight loss of righting reflex) while repeated cycles of 10mg/kg βAE treatment resulted in obvious motor and behavioral changes. Rats receiving 1mg/kg βAE had no brainstem lesions whereas some rats treated with 5mg/kg βAE and all rats treated with 10 mg/kg βAE had brainstem lesions. Brainstem lesions were observed after as few as five cycles and were characterized by gliosis, satellitosis and progressive necrosis in motor neurons of the trapezoid, vestibular, and olivary nuclei. This study shows that repeated treatment with clinically relevant doses of βAE causes motor deficits associated with brainstem damage in rodents and suggests that repeated treatment with βAE in children may elicit neurological damage.


Novel zebrafish behavioral assay to identify modifiers of the rapid, nongenomic stress response.
When vertebrates face acute stressors, their bodies rapidly undergo a repertoire of physiological and behavioral adaptations, which is termed the stress response. Rapid changes in heart rate and blood glucose levels occur via the interaction of glucocorticoids and their cognate receptors following hypothalamic-pituitary-adrenal axis activation. These physiological changes are observed within minutes of encountering a stressor and the rapid time domain rules out genomic responses that require gene expression changes. Although behavioral changes corresponding to physiological changes are commonly observed, it is not clearly understood to what extent hypothalamic-pituitary-adrenal axis activation dictates adaptive behavior. We hypothesized that rapid locomotor response to acute stressors in zebrafish requires hypothalamic-pituitary-interrenal (HPI) axis activation. In teleost fish, interrenal cells are functionally homologous to the adrenocortical layer. We derived eight frameshift mutants in genes involved in HPI axis function: two mutants in exon 2 of mc2r (adrenocorticotropic hormone receptor), five in exon 2 or 5 of nr3c1 (glucocorticoid receptor [GR]) and two in exon 2 of nr3c2 (mineralocorticoid receptor [MR]). Exposing larval zebrafish to mild environmental stressors, acute changes in salinity or light illumination, results in a rapid locomotor response. We show that this locomotor response requires a functioning HPI axis via the action of mc2r and the canonical GR encoded by nr3c1 gene, but not MR (nr3c2). Our rapid behavioral assay paradigm based on HPI axis biology can be used to screen for genetic and environmental modifiers of the hypothalamic-pituitary-adrenal axis and to investigate the effects of corticosteroids and their cognate receptor interactions on behavior.


Humoral immune responses to inactivated oil-emulsified Marek's disease vaccine.
When inactivated Md11/75C vaccine was inoculated into 1-day-old chickens, it stimulated antibodies detectable by enzyme-linked immunosorbent assay (at a titer of 6400) and indirect fluorescent antibody test (at a titer of 640), but lacking virus-neutralizing activity. Chickens passively inoculated with these antibodies were protected against bursal atrophy, weight loss, and early mortality when challenged with the virulent Md5 strain of Marek's disease virus (MDV). That led to the conclusion that virus-neutralizing activity is not a prerequisite for protection. In another experiment, antibody titers of adult chickens previously primed by exposure to live turkey herpesvirus and MDV did not increase after immunization with inactivated oil-emulsion MDV vaccines. This result provides little hope that Marek's disease can be controlled in progeny chickens by maternal immunity derived from hyperimmunized parents.


[Interaction of immobilization stress and food-getting learning].
The behavioral effects of emotional negative stress (immobilization) were studied in Wistar rats intact and those that had previous positive emotion experience. The food-getting learning has been chosen as positive emotion experience. Animals were trained in food pellet-reaching task by their preferred paw. It was shown that immobilization of intact rats leads to suppression of motor activity and increasing the duration of grooming. These effects indicate enhancement of passive-avoidance reactions. It was also shown that motor learning in group of rats with food reinforcement before immobilisation significantly reduces appearance of passive-avoidance reactions. It was found that immobilization stress does not inverse the initial direction of limb preference in majority of rats.


L1 endocytosis is controlled by a phosphorylation-dephosphorylation cycle stimulated by outside-in signaling by L1.
Dynamic regulation of the cell surface expression of adhesion molecules is an important mechanism for controlling neuronal growth cone motility and guidance. Clathrin-mediated vesicular internalization of L1 via the tyrosine-based endocytosis motif YRSL regulates adhesion and signaling by this Ig superfamily molecule. Here, we present evidence that tyrosine-1176 (Y1176) of the YRSL motif is phosphorylated in vivo. The nonreceptor tyrosine kinase (p60src) is implicated in L1-mediated neurite outgrowth, and we find that p60src phosphorylates Y1176 in vitro. Phosphorylation of Y1176 prevents L1 binding to AP-2, an adaptor required for clathrin-mediated internalization of L1. mAb 74-5H7 recognizes the sequence immediately NH2-terminal to the tyrosine-based motif and binds L1 only when Y1176 is dephosphorylated. 74-5H7 identifies a subset of L1 present at points of cell-cell contact and in vesicle-like structures that colocalize with an endocytosis marker. L1-L1 binding or L1 cross-linking induces a rapid increase in 74-5H7 immunoreactivity. Our data suggest a model in which homophilic binding or L1 cross-linking triggers transient dephosphorylation of the YRSL motif that makes L1 available for endocytosis. Thus, the regulation of L1 endocytosis through dephosphorylation of Y1176 is a critical regulatory point of L1-mediated adhesion and signaling.


Proteome changes induced by Pyrenophora tritici-repentis ToxA in both insensitive and sensitive wheat indicate senescence-like signaling.
Pyrenophora tritici-repentis is a phytopathogenic fungus which causes tan spot on wheat. Some races of P. tritici-repentis produce host-specific toxins which present symptoms of chlorosis or necrosis on susceptible wheat cultivars. One such toxin is Ptr ToxA, which enters mesophyll cells through a putative toxin-receptor and localizes with chloroplasts, ultimately causing damage and necrosis on leaves. These symptoms can occur even in the absence of the pathogen. Insensitive cultivars lack the receptor and Ptr ToxA cannot enter cells. The molecular mechanisms surrounding this plant-pathogen interaction are still largely unknown, although some details have begun to emerge. Using 2-D electrophoresis, fifteen protein changes were identified reproducibly in the leaf proteomes of a sensitive and an insensitive cultivar over three days after inoculation of purified Ptr ToxA. Functional analysis of the proteins indicated that senescence signals may be induced in the sensitive cultivar. In the insensitive cultivar proteins involved in some features of senescence inhibition were seen. Complementary responses at the biochemical level may be actively promoting a localized senescence-like response in sensitive wheat cultivars whilst actively inhibiting this response in insensitive cultivars. This is the first report of a biochemical response in an insensitive cultivar in this plant-pathogen interaction. Findings support the involvement of ethylene, and the activation of complementary pathways in sensitive versus insensitive wheat cultivars responding to Ptr ToxA. The nature of the system permits using purified toxin to mimic disease, which eliminates the pathogen proteome and ensures a synchronous response in inoculated leaves.


Invited review: Whey proteins as antioxidants and promoters of cellular antioxidant pathways.
Oxidative stress contributes to cell injury and aggravates several chronic diseases. Dietary antioxidants help the body to fight against free radicals and, therefore, avoid or reduce oxidative stress. Recently, proteins from milk whey liquid have been described as antioxidants. This review summarizes the evidence that whey products exhibit radical scavenging activity and reducing power. It examines the processing and treatment attempts to increase the antioxidant bioactivity and identifies 1 enzyme, subtilisin, which consistently produces the most potent whey fractions. The review compares whey from different milk sources and puts whey proteins in the context of other known food antioxidants. However, for efficacy, the antioxidant activity of whey proteins must not only survive processing, but also upper gut transit and arrival in the bloodstream, if whey products are to promote antioxidant levels in target organs. Studies reveal that direct cell exposure to whey samples increases intracellular antioxidants such as glutathione. However, the physiological relevance of these in vitro assays is questionable, and evidence is conflicting from dietary intervention trials, with both rats and humans, that whey products can boost cellular antioxidant biomarkers.


Degree of genetic liability for Alzheimer's disease associated with specific proteomic profiles in cerebrospinal fluid.
Genetic factors play a major role in Alzheimer's disease (AD) pathology, but biological mechanisms through which these factors contribute to AD remain elusive. Using a cerebrospinal fluid (CSF) proteomic approach, we examined associations between polygenic risk scores for AD (PGRS) and CSF proteomic profiles in 250 individuals with normal cognition, mild cognitive impairment, and AD-type dementia from the Alzheimer's Disease Neuroimaging Initiative. Out of 412 proteins, 201 were associated with PGRS. Hierarchical clustering analysis on proteins associated with PGRS at different single-nucleotide polymorphism p-value inclusion thresholds identified 3 clusters: (1) a protein cluster correlated with highly significant single-nucleotide polymorphisms, associated with amyloid-beta pathology and complement cascades; (2) a protein cluster associated with PGRS additionally including variants contributing to modest risk, involved in neural injury; (3) a protein cluster that also included less strongly associated variants, enriched with cytokine-cytokine interactions and cell adhesion molecules. These findings suggest that CSF protein levels reflect varying degrees of genetic liability for AD and may serve as a tool to investigate biological mechanisms in AD.


High-Resolution Microfluidic Paper-Based Analytical Devices for Sub-Microliter Sample Analysis.
This work demonstrates the fabrication of microfluidic paper-based analytical devices (µPADs) suitable for the analysis of sub-microliter sample volumes. The wax-printing approach widely used for the patterning of paper substrates has been adapted to obtain high-resolution microfluidic structures patterned in filter paper. This has been achieved by replacing the hot plate heating method conventionally used to melt printed wax features into paper by simple hot lamination. This patterning technique, in combination with the consideration of device geometry and the influence of cellulose fiber direction in filter paper, led to a model µPAD design with four microfluidic channels that can be filled with as low as 0.5 µL of liquid. Finally, the application to a colorimetric model assay targeting total protein concentrations is shown. Calibration curves for human serum albumin (HSA) were recorded from sub-microliter samples (0.8 µL), with tolerance against ±0.1 µL variations in the applied liquid volume.


Iodine Intake Estimation from the Consumption of Instant Noodles, Drinking Water and Household Salt in Indonesia.
The objective of this study was to assess the contribution of iodine intake from iodised household salt, iodised salt in instant noodles, and iodine in ground water in five regions of Indonesia. Secondary data analysis was performed using the 2013 Primary Health Research Survey, the 2014 Total Diet Study, and data from food industry research. Iodine intake was estimated among 2719 children, 10-12 years of age (SAC), 13,233 women of reproductive age (WRA), and 578 pregnant women (PW). Combined estimated iodine intake from the three stated sources met 78%, 70%, and 41% of iodine requirements for SAC, WRA and PW, respectively. Household salt iodine contributed about half of the iodine requirements for SAC (49%) and WRA (48%) and a quarter for PW (28%). The following variations were found: for population group, the percentage of estimated dietary iodine requirements met by instant noodle consumption was significantly higher among SAC; for region, estimated iodine intake was significantly higher from ground water for WRA in Java, and from household salt for SAC and WRA in Kalimantan and Java; and for household socio-economic status (SES), iodine intake from household salt was significantly higher in the highest SES households. Enforcement of clear implementing regulations for iodisation of household and food industry salt will promote optimal iodine intake among all population groups with different diets.


Distinctive characteristics and prognostic significance of interstitial pneumonia with autoimmune features in patients with chronic fibrosing interstitial pneumonia.
Interstitial lung diseases are heterogeneous, and patients with chronic fibrosing interstitial pneumonia (CFIP) often have clinical, serologic, and morphologic features suggestive but not diagnostic of connective tissue disease. Recently, the concept of interstitial pneumonia with autoimmune features (IPAF) has been proposed as a platform for such patients. However, the prognostic role of IPAF, including the cumulative incidence of acute exacerbations (AEs), is not fully clear. The aim of this study was to elucidate the clinical features and prognostic significance of IPAF. The clinical characteristics and prognostic relevance of a diagnosis of IPAF were retrospectively explored in 194 patients with CFIP, including 163 with idiopathic pulmonary fibrosis (IPF) and 31 with nonspecific interstitial pneumonia (NSIP), in our interstitial lung disease database. Sixteen percent of patients with CFIP (8% of IPF, 61% of NSIP) met the criteria for IPAF. Patients with IPAF were significantly younger and included a higher proportion of women, never-smokers, and patients with NSIP than those without IPAF. The morphologic domain was the most common in patients with IPAF (97%), followed by the serologic domain (72%) and clinical domain (53%). CFIP patients with IPAF had a more favorable prognosis with regard to overall survival (OS; P < 0.001, log-rank test) and incidence of AEs (P = 0.029, Gray's test) than those without IPAF. In the subgroup analysis, NSIP patients with IPAF had significantly better survival than those without IPAF (P = 0.031, log-rank test), and IPF patients with IPAF tended to have better OS than those without IPAF (P = 0.092, log-rank test). However, there were no significant differences in the incidence of AEs between patients with IPAF and those without IPAF in the IPF and NSIP subgroups. Furthermore, fulfilment of the IPAF criteria was an independent predictor of OS (hazard ratio (HR) 0.127; 95% confidence interval (CI) 0.017-0.952; P = 0.045) and incidence of AEs (HR 0.225: 95% CI 0.054-0.937; P = 0.040). A diagnosis of IPAF might predict a favorable prognosis and less risk of AEs in patients with CFIP.


"This has changed me to be a better mum": A qualitative study exploring how the Australian Nurse-Family Partnership Program contributes to the development of First Nations women's self-efficacy.
The Australian Nurse-Family Partnership Program is based on the Nurse-Family Partnership program from the United States, which was designed to support first-time mothers experiencing social and economic disadvantage from early in pregnancy until their child's second birthday. International trials have demonstrated this program measurably improves family environment, maternal competencies, and child development. The Australian program has been tailored for mothers having a First Nations baby. This study aimed to understand how the program impacts self-efficacy using a qualitative interpretive approach. The study took place in two sites within one Aboriginal Community Controlled Health Service in Meanjin (Brisbane), Australia. Twenty-nine participants were interviewed: first-time mothers having a First Nations baby who had accessed the program (n = 26), their family members (n = 1), and First Nations Elders (n = 2). Interviews were conducted either face-to-face or by telephone, using a yarning tool and method, to explore women's experiences and perceptions. Yarns were analysed using reflexive thematic analysis. Three main themes were generated: 1) sustaining connections and relationships; 2) developing self-belief and personal skills; and 3) achieving transformation and growth. We interpret that when the program facilitates the development of culturally safe relationships with staff and peers, it enables behaviour change, skill development, personal goal setting and achievement, leading to self-efficacy. Located within a community-controlled health service, the program can foster cultural connection, peer support and access to health and social services; all contributing to self-efficacy. We recommend the program indicators are strengthened to reflect these findings and enable monitoring and reporting of activities that facilitate self-efficacy, growth, and empowerment.


Familial juvenile hyperuricemic nephropathy: localization of the gene on chromosome 16p11.2-and evidence for genetic heterogeneity.
Familial juvenile hyperuricemic nephropathy (FJHN), is an autosomal dominant renal disease characterized by juvenile onset of hyperuricemia, gouty arthritis, and progressive renal failure at an early age. Using a genomewide linkage analysis in three Czech affected families, we have identified, on chromosome 16p11.2, a locus for FJHN and have found evidence for genetic heterogeneity and reduced penetrance of the disease. The maximum two-point LOD score calculated with allowance for heterogeneity (HLOD) was 4.70, obtained at recombination fraction 0, with marker D16S3036; multipoint linkage analysis yielded a maximum HLOD score of 4.76 at the same location. Haplotype analysis defined a 10-cM candidate region between flanking markers D16S501 and D16S3113, exhibiting crossover events with the disease locus. The candidate interval contains several genes expressed in the kidney, two of which-uromodulin and NADP-regulated thyroid-hormone-binding protein-represent promising candidates for further analysis.


FMR1 Low Zone CGG Repeats: Phenotypic Associations in the Context of Parenting Stress.
The FMR1 gene on the X chromosome has varying numbers of CGG repeats. The modal number is 30, and expansion to >200 results in fragile X syndrome, but the copy number extends down to 6. Past research suggests that individuals whose CGGs are in the "low zone" (LZ; defined here as ≤ 25 CGGs) may be more environmentally-reactive than those with normal range repeats (26-40 CGGs)-a gene x environment interaction. Using a population-based DNA biobank, in our primary analysis we compared 96 mothers with LZ CGG repeats on both alleles to 280 mothers who had CGG repeats in the normal range. Secondarily, we conducted parallel analyses on fathers. We investigated how parents in these two CGG repeat categories differentially responded to stress, defined as parenting a child with disabilities. Significant gene x environment interactions indicated that LZ mothers who had children with disabilities had greater limitations (in executive functioning, depression, anxiety, daily health symptoms, and balance) than LZ mothers whose children did not have disabilities. In contrast, mothers with normal-range CGG repeats did not differ based on stress exposure. For fathers, a similar pattern was evident for one phenotype only (hand tremors). Although on average LZ CGGs are not associated with compromised functioning, the average masks differential response to the environment.


Changing the paradigm in the treatment of platinum-sensitive recurrent ovarian cancer: from platinum doublets to nonplatinum doublets and adding antiangiogenesis compounds.
The optimal treatment for women with recurrent epithelial ovarian cancer is evolving. The objective of this review is to outline the transition away from platinum doublets toward nonplatinum combinations and review emerging data on antiangiogenesis therapy in this setting. Recently published and presented data as well as ongoing clinical trials are discussed. Current clinical practice largely harmonizes with a paradigm that outlines a treatment algorithm for recurrent ovarian cancer based on the duration of platinum-free exposure. In this model, patients whose penultimate platinum compound exposure (platinum-free interval [PFI]) is longer than 6 months are generally offered a platinum agent or a platinum-containing doublet; those with a shorter interval are usually treated with a single nonplatinum agent. This is based on the simple contention that better clinical outcomes will be realized with platinum in those deemed platinum sensitive (PFI >6 months). However, it is becoming clear from various phase II and phase III clinical studies that the performance of many nonplatinum chemotherapeutic agents is also influenced by this parameter (PFI). Indeed, although definitive comparisons of nonplatinum drugs to novel cytotoxic agents are lacking, the clinical activity of these compounds might approach or exceed that of platinum agents. Although recognized by clinicians, the dichotomy that determines therapy based on PFI has not been formally accepted by the US Food and Drug Administration in all cases of drug labeling. For instance, whereas the combination of gemcitabine and carboplatin is now approved for patients with platinum-sensitive recurrent ovarian cancer, traditionally used platinum-resistant (PFI <6 months) agents such as topotecan and pegylated liposomal doxorubicin are also approved by the US Food and Drug Administration as single agents in platinum-sensitive patients. Furthermore, the nonplatinum doublet pegylated liposomal doxorubicin and trabectedin has recently documented comparable activity to platinum combinations among patients with a PFI of longer than 6 months. To that end, the most prolific developmental therapeutics arena in ovarian cancer is biologically targeted therapy, particularly angiogenesis inhibitors. Although it is unknown if the clinical activity from these new agents will respect the chemotherapy-sensitive dichotomy, it is clear that they have the potential to augment efficacy, possibly in both traditionally chemosensitive and chemoresistant phenotypes. The term platinum sensitive should probably be replaced by chemotherapy sensitive, particularly as new nonplatinum agents and combinations are identified as active in this setting. Nonplatinum doublets are effective in treating platinum-sensitive recurrent disease, and adding antiangiogenesis agents to these combinations is a research priority.


Cryptosporulation in Kurthia spp. forces a rethinking of asporogenesis in Firmicutes.
Endosporulation is a complex morphophysiological process resulting in a more resistant cellular structure that is produced within the mother cell and is called endospore. Endosporulation evolved in the common ancestor of Firmicutes, but it is lost in descendant lineages classified as asporogenic. While Kurthia spp. is considered to comprise only asporogenic species, we show here that strain 11kri321, which was isolated from an oligotrophic geothermal reservoir, produces phase-bright spore-like structures. Phylogenomics of strain 11kri321 and other Kurthia strains reveals little similarity to genetic determinants of sporulation known from endosporulating Bacilli. However, morphological hallmarks of endosporulation were observed in two of the four Kurthia strains tested, resulting in spore-like structures (cryptospores). In contrast to classic endospores, these cryptospores did not protect against heat or UV damage and successive sub-culturing led to the loss of the cryptosporulating phenotype. Our findings imply that a cryptosporulation phenotype may have been prevalent and subsequently lost by laboratory culturing in other Firmicutes currently considered as asporogenic. Cryptosporulation might thus represent an ancestral but unstable and adaptive developmental state in Firmicutes that is under selection under harsh environmental conditions.


Fear conditioning, safety learning, and sleep in humans.
Fear conditioning is considered an animal model of post-traumatic stress disorder. Such models have shown fear conditioning disrupts subsequent rapid eye movement sleep (REM). Here, we provide a translation of these models into humans. Using the fear potentiated startle (FPS) procedure, we examined the effects of fear conditioning and safety signal learning on subsequent REM sleep in healthy adults. We also examined the effects of changes in REM sleep on retention of fear and safety learning. Participants (n = 42 normal controls) spent 3 consecutive nights in the laboratory. The first was an adaptation night. Following the second night, we administered a FPS procedure that included pairing a wrist shock with a threat signal and a safety signal never paired with a shock. The next day, we administered the FPS procedure again, with no wrist shocks to any stimulus, to measure retention of fear and safety. Canonical correlations assessed the relationship between FPS response and REM sleep. Results demonstrated that increased safety signal learning during the initial acquisition phase was associated with increased REM sleep consolidation that night, with 28.4% of the variance in increased REM sleep consolidation from baseline accounted for by safety signal learning. Overnight REM sleep was, in turn, related to overnight retention of fear and safety learning, with 22.5% of the variance in startle retention accounted for by REM sleep. These data suggest that sleep difficulties, specifically REM sleep fragmentation, may play a mechanistic role in post-traumatic stress disorder via an influence on safety signal learning and/or threat-safety discrimination.


Protective-antigen (PA) based anthrax vaccines confer protection against inhalation anthrax by precluding the establishment of a systemic infection.
An intense effort has been launched to develop improved anthrax vaccines that confer rapid, long lasting protection preferably with an extended stability profile amenable for stockpiling. Protective antigen (PA)-based vaccines are most favored as immune responses directed against PA are singularly protective, although the actual protective mechanism remains to be unraveled. Herein we show that contrary to the prevailing view, an efficacious PA-based vaccine confers protection against inhalation anthrax by preventing the establishment of a toxin-releasing systemic infection. Equally importantly, antibodies measured by the in vitro lethal toxin neutralization activity assay (TNA) that is considered as a reliable correlate of protection, especially for PA protein-based vaccines adjuvanted with aluminum salts appear to be not absolutely essential for this protective immune response.


Better results in consolidation of hallux metatarsophalangeal arthrodesis with dorsal plate and interfragmentary compression screw.
The first metatarsophalangeal joint arthrodesis is indicated for the treatment of various pathologies as a technique to reduce pain and improve the support of the first radius. Numerous surgical techniques and fixation methods have been described, with the combination of a dorsal plate and an interfragmentary screw being the one that has shown to be the most stable construct in biomechanical studies Our aim is to analyze the radiological results after metatarsophalangeal arthrodesis of the hallux using a dorsal plate associated or not with an interfragmentary screw. The differences in terms of consolidation rates and complications in patients diagnosed with hallux rigidus, hallux valgus, hallux varus and failure of previous surgeries were evaluated. A retrospective cohort study of 55 patients with a mean age of 65.10 years in whom a dorsal plate was used was performed. Patients were divided into two groups depending on whether or not an interfragmentary screw was used. The minimum follow-up was 6 months after surgery. The assessment of the pre and postoperative radiological results was based on the variation of the hallux angle, the intermetatarsal angle and the dorsal metatarsophalangeal angle of the hallux, as well as the cases of nonunion identified in each study group. The radiological results, statistically significant differences (p<0.05) were only found in the dorsal metatarsophalangeal angle between both study groups. No statistically significant differences were found regarding the radiological evaluation of the pre and postoperative hallux angle and intermetatarsal angle. An equal decrease of each angles was observed in both study groups. Regarding the consolidation rate, statistically significant differences (p<0.05) were found between group A, which associated an interfragmentary screw, presenting a consolidation rate of 92%, and group B, which did not associate an interfragmentary screw, and that presented a union rate of 63%. Hallux metatarsophalangeal arthrodesis of the hallux with a dorsal plate and interfragmentary screw show best results regarding consolidation rate and complications compared to those cases in which an interfragmentary screw was not used.


A frequent 1085delC/insGAAG mutation in the RDH5 gene in Japanese patients with fundus albipunctatus.
To identify the frequency of a mutation of the RDH5 gene in Japanese patients with hereditary retinal degeneration and to characterize clinical findings for the patients associated with a 1085delC/insGAAG mutation in the RDH5 gene. Mutation screening by single-strand conformation polymorphism was performed on 6 patients with fundus albipunctatus and 150 patients with autosomal recessive retinitis pigmentosa. The DNA fragment that showed abnormal mobility on SSCP was then sequenced. Clinical features were characterized by visual acuity, slit-lamp biomicroscopy, electroretinography, fluorescein angiography, kinetic visual field testing, and dark adaptometry. A novel 1085delC/insGAAG mutation in the RDH5 gene was identified in all 6 patients, from 4 unrelated families with fundus albipunctatus. The ophthalmic findings of each affected member were very similar, which may provide the natural course of the phenotype produced by the 1085delC/insGAAG mutation. A homozygous1085delC/insGAAG mutation in the RDH5 gene produces fundus albipunctatus in Japanese patients. These findings suggest that this mutation was a founder effect in Japanese patients with fundus albipunctatus.


DNA sequences of random origin as probes of Escherichia coli promoter architecture.
In order to better understand the role of the -35 sequence motif in transcription initiation by Escherichia coli sigma 70 RNA polymerase holoenzyme, -35 promoter elements of limited nucleotide composition have been selected from random DNA sequences. Functional promoter elements have been identified that are composed of just two nucleotide species (A,T; G,C; T,C; and T,G) and one nucleotide species (poly(dT)). From this study of 81 promoter mutations, two conclusions can be made. First, given a population of random DNA sequences, there exist sequences that form stronger -35 promoter elements than the consensus sequence. These sequences lack some features of the consensus sequence and reveal unexpected homology in the ordinarily nonconserved spacer sequence. Second, the sequence requirements at the -35 site may suggest a possible role for DNA secondary structure in the recognition of promoters by RNA polymerase.


Mitochondrial Hspa9/Mortalin regulates erythroid differentiation via iron-sulfur cluster assembly.
Mitochondrial iron-sulfur cluster (ISC) biogenesis provides iron-sulfur cofactors to several mitochondrial proteins, but the extent to which ISC biogenesis regulates hematopoiesis has been unclear. The blood disease Myelodysplastic syndrome (MDS) is characterized by ineffective hematopoiesis, and the disease overlaps with the gene Hspa9/Mortalin in multiple ways: the HSPA9 locus maps to 5q31.2 that is frequently deleted in human MDS; mutant Hspa9 causes zebrafish MDS; and Hspa9 knockdown mice have decreased hematopoiesis. We show here that HSPA9 functions in mitochondrial ISC biogenesis, and is required for erythroid differentiation. HSPA9 interacts with and stabilizes the mitochondrial ISC biogenesis proteins frataxin, Nfs1, ISCU, and Nfu. MDS-causing mutations in HSPA9 protein change its interactions with ISC biogenesis proteins. Depletion of HSPA9 decreases aconitase activity, which requires an ISC at its active site, but not that of the non-ISC requiring malate dehydrogenase, and increases IRP1 binding activity. In erythroid cell lines, Hspa9 depletion inhibited erythroid differentiation, post-transcriptionally regulating the expression of Alas2 and FeCH, as expected through known ISC control of the IRE response elements in these genes. By contrast, the Alas2 open reading frame rescued the Hspa9-dependent defect in erythroid differentiation, but not when uncoupled from its 5'-IRE sequence. Thus, Hspa9 depletion causes a mitochondrial ISC deficit, altering IRP1-IRE binding and FeCH stability, which consequently inhibits Alas2 translation, heme synthesis, and erythroid differentiation, i.e.: Hspa9->ISC->IRP/IRE->Alas2->heme synthesis->erythroid differentiation. Thus Hspa9 regulates erythroid differentiation through ISC cluster assembly, providing a pathophysiological mechanism for an MDS subtype characterized by HSPA9 haploinsufficiency, and suggests hemin and other pharmacological stimulators of ISC synthesis as potential routes to therapy.


Solid-state energy-dispersion spectrometer for electron-microprobe x-ray analysis.
Improved lithium-drifted silicon solid-state detectors allow detection and energy dispersion of x-rays of about 3 to 30 kiloelectron volts in the electron-microprobe x-ray analyzer. Energy resolution is sufficient to separate peaks of characteristic x-rays of elements adjacent in the periodic system at atomic number 20 and higher. The detected x-ray spectrum emitted from an unknown sample can be recorded with a multichannel analyzer in approximately 60 seconds.


Large vestibular schwannomas: long-term outcomes after stereotactic radiosurgery.
Stereotactic radiosurgery (SRS) is an established treatment option of small/medium-sized vestibular schwannomas (VSs). Concerning management of the large VSs, primary SRS remains a controversial option. Our retrospective study analyzes long-term radiological and clinical outcomes of SRS in large VSs. We retrospectively analyzed 73 patients with single large VS, treated with SRS. Inclusion criteria were: tumor volume >4 cm3, follow-up >2 years, radiological (3D-volumetric studies) and clinical follow-up. SRS was either primary (94.5%) or secondary (5.5%) treatment. The median marginal dose (50%-isodose line) was 12Gy (11.5-12Gy). Fisher exact test, t-test, ANOVA, Kaplan-Meier and Cox regression models were performed when appropriate RESULTS: The median follow-up was 5.5 years. The median VS volume at SRS was 6.5 cm3 (range 4-14.2 cm3). The tumor control rates assessed from Kaplan-Meier curve were 88.3%, 82.4% and 74.7% 5.8 and 10 years after SRS, respectively. Tumor shrinkage was observed in 83.6% of patients (n=61), unchanged volume in 4.1% patients (n=3) and progression in 12.3% (n=9). The median tumor volume significantly decreased to 4.0 cm3, measured at 5-year follow-up (p<0.0001). Large cystic VSs responded better to SRS then homogeneous. Pre-SRS serviceable hearing was present in 37% of patients; 55% of these had hearing preserved after treatment. After SRS, new facial palsy (House-Brackmann gr. III-VI) appeared in 4.1% of patients; 9.6% of patients had transient brainstem/cranial nerves edema. For tumor progression, 8.2% of patients underwent resection, 2.8% of patients repeated SRS. Our results are showing that SRS might be safe and effective primary treatment even in large VSs. However, long-term tumor control rates are lower in comparison with small/medium-sized VSs. Thus, closer follow-up should be applied.


Penetration of host cell lines by bacteria. Characteristics of the process of intracellular bacterial infection.
A model which describes the characteristics of the penetration of the cells by bacteria is presented. Since the process of invasion is preceded necessarily by the step in which the bacteria adhere to the cells, the proposed model is based on the expressions previously derived for the process of adhesion, which allow us to determine the number of attached bacteria under different conditions. Thus, the model considers that invasion occurs irreversibly from attached bacteria to specific receptors located on the cell surface with a rate coefficient = ki so that the invasive capacity in a given bacterium-host cell system is mainly determined by the value of this coefficient. Once internalized, the bacteria can follow three different time courses, namely: 1) intracellular growth is hindered so that the bacteria remain in stationary phase, 2) there is a lag phase during which the bacteria stay in stationary phase before they are able to grow exponentially with a rate coefficient = kc, and 3) the bacteria exhibit a growth exponential phase as they enter the cells. In turn, the time course followed by extracellular bacteria also has a decisive influence on the process of invasion and, in this regard, unbound bacteria are considered either in stationary or in exponential phase. Expressions for these different situations have been derived, and from them, procedures to determine the levels of bacterial infection and for quantitative invasive data analysis are presented.


Characterization of an aspartate aminotransferase encoded by YPO0623 with frequent nonsense mutations in Yersinia pestis.
Yersinia pestis, the causative agent of plague, is a genetically monomorphic bacterial pathogen that evolved from Yersinia pseudotuberculosis approximately 7,400 years ago. We observed unusually frequent mutations in Y. pestis YPO0623, mostly resulting in protein translation termination, which implies a strong natural selection. These mutations were found in all phylogenetic lineages of Y. pestis, and there was no apparent pattern in the spatial distribution of the mutant strains. Based on these findings, we aimed to investigate the biological function of YPO0623 and the reasons for its frequent mutation in Y. pestis. Our in vitro and in vivo assays revealed that the deletion of YPO0623 enhanced the growth of Y. pestis in nutrient-rich environments and led to increased tolerance to heat and cold shocks. With RNA-seq analysis, we also discovered that the deletion of YPO0623 resulted in the upregulation of genes associated with the type VI secretion system (T6SS) at 26°C, which probably plays a crucial role in the response of Y. pestis to environment fluctuations. Furthermore, bioinformatic analysis showed that YPO0623 has high homology with a PLP-dependent aspartate aminotransferase in Salmonella enterica, and the enzyme activity assays confirmed its aspartate aminotransferase activity. However, the enzyme activity of YPO0623 was significantly lower than that in other bacteria. These observations provide some insights into the underlying reasons for the high-frequency nonsense mutations in YPO0623, and further investigations are needed to determine the exact mechanism.