A quantitative in vivo mouse model used to assay inhibitors of tumor-induced angiogenesis.
An in vivo model of tumor-induced angiogenesis was used to monitor two known inhibitors of angiogenesis, protamine sulfate and the steroid tetrahydro S. Tumor cells entrapped in alginate beads were injected s.c. into mice. Blood vessel induction was measured by two quantitative methods: measurement of hemoglobin at the alginate pellet site, and pooling of radiolabeled RBC to the alginate pellet site. The two methods gave parallel results. Tetrahydro S with or without heparin inhibited blood vessel growth by 50%, and protamine sulfate inhibited blood vessel growth by 85%. These results were supported by gross morphology and histological analysis of the alginate pellet site.


Age-related changes in wavelength discrimination.
Wavelength discrimination functions (420 to 620-650 nm) were measured for four younger (mean 30.9 years) and four older (mean 72.5 years) observers. Stimuli consisted of individually determined isoluminant monochromatic lights (10 Td) presented in each half of a 2 degrees circular bipartite field with use of a Maxwellian-view optical system. A spatial two-alternative forced-choice method was used in combination with a staircase procedure to determine discrimination thresholds across the spectrum. Small but consistent elevations in discrimination thresholds were found for older compared with younger observers. Because the retinal illuminance of the stimuli was equated across all observers, these age-related losses in discrimination are attributable to neural changes. Analyses of these data reveal a significant change in Weber fraction across adulthood for a chromatically opponent pathway receiving primarily antagonistic signals from middle-wavelength-sensitive and long-wavelength-sensitive cones but not for a short-wavelength-sensitive cone pathway.


Interoceptive technologies for psychiatric interventions: From diagnosis to clinical applications.
Interoception-the perception of internal bodily signals-has emerged as an area of interest due to its implications in emotion and the prevalence of dysfunctional interoceptive processes across psychopathological conditions. Despite the importance of interoception in cognitive neuroscience and psychiatry, its experimental manipulation remains technically challenging. This is due to the invasive nature of existing methods, the limitation of self-report and unimodal measures of interoception, and the absence of standardized approaches across disparate fields. This article integrates diverse research efforts from psychology, physiology, psychiatry, and engineering to address this oversight. Following a general introduction to the neurophysiology of interoception as hierarchical predictive processing, we review the existing paradigms for manipulating interoception (e.g., interoceptive modulation), their underlying mechanisms (e.g., interoceptive conditioning), and clinical applications (e.g., interoceptive exposure). We suggest a classification for interoceptive technologies and discuss their potential for diagnosing and treating mental health disorders. Despite promising results, considerable work is still needed to develop standardized, validated measures of interoceptive function across domains and before these technologies can translate safely and effectively to clinical settings.


Diffusion and ADC-map images detect ongoing demyelination on subcortical white matter in an adult metachromatic leukodystrophy patient with autoimmune Hashimoto thyroiditis.
Adult-onset metachromatic leukodystrophy (MLD) often shows schizophrenia- or encephalopathy-like symptoms at an early stage, such as behavioural abnormalities, cognitive impairment, mood disorders and hallucinations. The authors report the case of an adult woman with MLD who had been given antipsychotic medication for schizophrenia. In the differential diagnosis, screening of auto-antibodies was important for ruling out other encephalopathies as she had a euthyroid Hashimoto thyroiditis. Diagnosis was based the results of MRI, nerve conduction velocity, sensory evoked potential, motor evoked potential, lysosomal enzyme activity and gene analysis studies. Brain MRI showed diffuse demyelination spreading from the deep white matter to subcortical area as high signals at the edges of these lesions in diffusion and apparent diffusion coefficient-map images with the U-fibres conserved. The authors diagnosed adult-onset MLD coexisting with euthyroid autoimmune Hashimoto thyroiditis.


Comparison of density functionals for reactions of sulfur ylides with aldehydes and olefins.
The reactions of a model sulfur ylide with formaldehyde and 1,1-dicianoethylene, leading to the formation of an epoxyde and a cyclopropane, respectively, have been studied using different computational methods, and the results have been compared to those obtained with the CBS-QB3 method. The second step of these reactions presents transition states similar to that of an SN2 reaction. Depending on the degree of electron delocalization at the transition state, a different amount of exact exchange is necessary in the exchange functional to obtain accurate energy barriers. This amount is larger for the reaction of formaldehyde, in which the transition state is more delocalized, than for the reaction of 1,1-dicianoethylene. Similar results have been obtained for symmetric and non-symmetric SN2 reactions. The calculation of the reaction path has shown that the error relative to CBS-QB3 tends to increase when approaching the transition state. Among the different computational methods, PBE1PBE is the one to provide the most accurate energy barriers and reaction energies, whereas BB1K leads to the best results for the reaction path before the transition state.


When the ward is the patient: Using the PRISM protocol to understand and reduce violence in an inpatient intellectual disability setting.
Violence in inpatient settings is recognised as a worldwide issue, with inpatient intellectual disability services having higher rates than other mental health settings. Violence results in injury and illness, lack of confidence in the organisation and staff burnout. These combined effects have a negative impact on the ability of services to provide therapeutic environments. Attempts to manage violence tend to focus on the individual. This is only one part of the solution. Situational risk factors for violence within secure settings can be identified and modified to reduce violence rates. The role of situational risk factors in rates of institutional violence requires consideration in addition to individual patient risk assessment and management. To discuss an illustrative case study of a ward experiencing high levels of violence, resulting in staff burnout and low morale. To describe how assessing and addressing the relevant situational risk factors led to demonstrable reductions in violence and improvements in the service provided. This case study demonstrates the practical application of assessing and managing situational risk factors for violence in a locked intellectual disability ward in order to effectively reduce levels of violent incidents. A description of the violence and critical issues faced by the ward is provided, noting key elements of the timeline of events and the application of the Promoting Risk Intervention by Situational Management (PRISM) process to address these. The PRISM protocol enabled a comprehensive assessment and understanding of situational factors relevant to the violence rates in the ward. The recommendations following the analysis enabled the management team to identify areas for immediate and long-term action. Practical steps to address issues such as improving soundproofing were implemented quickly whilst other issues required changes over the longer term. Significantly, reduced levels of violence were observed within a 2-month period of immediate steps being taken, with further gains achieved over the longer term. This paper provides the only example of the use of the PRISM protocol in an intellectual disability inpatient setting. The reduction in violence seen in this unit provides 'real world' evidence that addressing situational factors for institutional violence can be effective in intellectual disability inpatient settings, with beneficial outcomes for patients and staff. Whilst individual patient risk assessment and risk management are key components of forensic mental health care, situational risk factors for violence are highly relevant to rates of violence within secure care settings. Structured assessment of situational risk factors should be considered when inpatient settings experience persistent high levels of violence. Further research on the impact of assessing situational risk factors in forensic mental health settings (including intellectual disability services) would help to develop the existing evidence base.


Natural Products from Cyanobacteria: Focus on Beneficial Activities.
Cyanobacteria are photosynthetic microorganisms that colonize diverse environments worldwide, ranging from ocean to freshwaters, soils, and extreme environments. Their adaptation capacities and the diversity of natural products that they synthesize, support cyanobacterial success in colonization of their respective ecological niches. Although cyanobacteria are well-known for their toxin production and their relative deleterious consequences, they also produce a large variety of molecules that exhibit beneficial properties with high potential in various fields (e.g., a synthetic analog of dolastatin 10 is used against Hodgkin's lymphoma). The present review focuses on the beneficial activities of cyanobacterial molecules described so far. Based on an analysis of 670 papers, it appears that more than 90 genera of cyanobacteria have been observed to produce compounds with potentially beneficial activities in which most of them belong to the orders Oscillatoriales, Nostocales, Chroococcales, and Synechococcales. The rest of the cyanobacterial orders (i.e., Pleurocapsales, Chroococcidiopsales, and Gloeobacterales) remain poorly explored in terms of their molecular diversity and relative bioactivity. The diverse cyanobacterial metabolites possessing beneficial bioactivities belong to 10 different chemical classes (alkaloids, depsipeptides, lipopeptides, macrolides/lactones, peptides, terpenes, polysaccharides, lipids, polyketides, and others) that exhibit 14 major kinds of bioactivity. However, no direct relationship between the chemical class and the respective bioactivity of these molecules has been demonstrated. We further selected and specifically described 47 molecule families according to their respective bioactivities and their potential uses in pharmacology, cosmetology, agriculture, or other specific fields of interest. With this up-to-date review, we attempt to present new perspectives for the rational discovery of novel cyanobacterial metabolites with beneficial bioactivity.


Structural characterization of SARS-CoV-2 dimeric ORF9b reveals potential fold-switching trigger mechanism.
The constant emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants indicates the evolution and adaptation of the virus. Enhanced innate immune evasion through increased expression of viral antagonist proteins, including ORF9b, contributes to the improved transmission of the Alpha variant; hence, more attention should be paid to these viral proteins. ORF9b is an accessory protein that suppresses innate immunity via a monomer conformation by binding to Tom70. Here, we solved the dimeric structure of SARS-CoV-2 ORF9b with a long hydrophobic tunnel containing a lipid molecule that is crucial for the dimeric conformation and determined the specific lipid ligands as monoglycerides by conducting a liquid chromatography with tandem mass spectrometry analysis, suggesting an important role in the viral life cycle. Notably, a long intertwined loop accessible for host factor binding was observed in the structure. Eight phosphorylated residues in ORF9b were identified, and residues S50 and S53 were found to contribute to the stabilization of dimeric ORF9b. Additionally, we proposed a model of multifunctional ORF9b with a distinct conformation, suggesting that ORF9b is a fold-switching protein, while both lipids and phosphorylation contribute to the switching. Specifically, the ORF9b monomer interacts with Tom70 to suppress the innate immune response, whereas the ORF9b dimer binds to the membrane involving mature virion assembly. Our results provide a better understanding of the multiple functions of ORF9b.


Current status of pharmacovigilance regulatory structures, processes, and outcomes in the Asia-Pacific region: Survey results from 15 countries.
Regulatory discrepancies may exist in pharmacovigilance (PV) structure, process, and outcome status worldwide. Our study's objective was to survey the current status of PV in each regulatory body in the Asia-Pacific Economic Cooperation (APEC) region. A modified questionnaire was sent to the PV team heads of 21 PV agencies based in the APEC countries, between June 28 and September 12, 2017, to gather information on the structure, process, and outcome of PV status in these countries. Of the 21 APEC countries, 15 responded. We found harmonized laws and regulations for general PV and risk management systems. However, variations were found in PV structure: for example, 11 out of 15 countries had national regulatory representatives responsible for PV in pharmaceutical companies, while four did not. For PV process, discrepancies were also found in the source type of adverse drug reaction (ADR) reports and reporting of medication errors and therapeutic ineffectiveness in cumulative ADR reports. With respect to PV outcomes, among countries that performed active surveillance, the United States of America was more active, with hundreds of projects including additional pharmacoepidemiological studies etc. Among the nine countries that responded, Japan had the greatest number of product label changes followed by Taiwan, Malaysia, and Korea. We have identified substantial variations in the structures, processes, and outcomes of PV status among the countries of the APEC region. Therefore, efforts to reduce variations in the PV administration and regulation are warranted for harmonization of PV within the APEC region.


Diagnosis and management of non-calcium-containing stones in the pediatric population.
Compared to adults, urolithiasis is less common in children, with a definite rise in incidence, especially among young adults (Tasian et al. in Clin J Am Soc Nephrol 11:488, 2016). In the last 25 years, the incidence in children has increased by approximately 6-10% annually, for reasons still unknown, with an associated significant increase in related health care-related expenditures (Hyams and Matlaga in Transl Androl Urol 3(3):278-83, 2014). It has been shown that there is twice as high a risk of chronic kidney disease (CKD) or end stage renal disease (ESRD) in stone formers compared to non-stone formers (Tasian et al. 2016). While calcium-containing stones are by far the most common category of stone encountered in both children and adults, non-calcium stones are more common in children than adults and have been shown in several studies to be associated with greater morbidity and lower renal function than calcium stones (Issler et al. in BMC Nephrol 18(1):136, 2017; Gambaro et al. in J Urol 198:268-273, 2017). This could be related to the challenges in the management of non-calcium-containing stones due to associated infection or metabolic derangements, further leading to recurrence and loss of renal function. There is currently a gap in our understanding of how to appropriately and effectively encounter and manage patients with non-calcium-containing stones, as such cases are encountered less frequently. Identification of stone composition and appropriate management is very important to reduce serious complications and recurrence, especially in non-calcium stones. We present a review of diagnosis and management of non-calcium-containing stones in the pediatric population, in hopes of providing more clarity to providers and promoting a consideration of non-calcium stone composition with all children presenting with urolithiasis.


Development of 3-acetylindole derivatives that selectively target BRPF1 as new inhibitors of receptor activator of NF-κB ligand (RANKL)-Induced osteoclastogenesis.
Bromodomain and PHD finger-containing (BRPF) proteins function as epigenetic readers that specifically recognize acetylated lysine residues on histone tails. The acetyl-lysine binding pocket of BRPF has emerged as an attractive target for the development of protein interaction inhibitors owing to its potential druggability. In this study, we identified 3-acetylindoles as bone antiresorptive agents with a novel scaffold by performing structure-based virtual screening and hit optimization. Among those derivatives, compound 18 exhibited potent and selective inhibitory activities against BRPF1B (IC50 = 102 nM) as well as outstanding inhibitory activity against osteoclastogenesis (73.8% @ 1 μM) and differentiation (IC50 = 0.19 μM) without cytotoxicity. Besides, cellular mechanism assays demonstrated that compound 18 exhibited a strong bone antiresorptive effect by modulating the RANKL/RANK/NFATc1 pathway. Structural and functional studies on BRPF1 inhibitors aid in making advances to understand the epigenetic mechanisms of bone cell development and create innovative therapeutics for treating bone metastases from solid tumors and other bone erosive diseases.


Phosphorylation of p53 at the casein kinase II site selectively regulates p53-dependent transcriptional repression but not transactivation.
The p53 tumour suppressor protein is a potent transcription factor which plays a central role in the defence of cells against DNA damage and the propagation of malignant clones. We have previously shown that phosphorylation of serine 386 in mouse p53 by the growth- associated protein kinase, casein kinase II (CKII), plays an important role in the ability of p53 to block the proliferation of drug-resistant colonies. In this paper we show that blocking phosphorylation of serine 386 through an alanine substitution, or placing a constitutive negative charge at this position in the form of aspartate, had no significant influence on p53-dependent transcriptional activation of a promoter containing 13 copies of a p53 consensus binding sequence, or of the p21WAF1 promoter which is a natural target for p53. In contrast, the alanine mutant showed a weak reduction in the ability of p53 to repress expression from the c-fos promoter, which is a target for p53-dependent repression in vivo. Strikingly, when the repression of the SV40 early promoter was examined, a reduction in the repression capacity of up to 5-fold was observed. Moreover, repression of the SV40 promoter could be partially restored by aspartic acid substitution at the phosphorylation site. These data indicate that phosphorylation at a specific C-terminal site can selectively regulate p53-dependent repression, but not transactivation.


Prediction of tumor purity from gene expression data using machine learning.
Bulk tumor samples used for high-throughput molecular profiling are often an admixture of cancer cells and non-cancerous cells, which include immune and stromal cells. The mixed composition can confound the analysis and affect the biological interpretation of the results, and thus, accurate prediction of tumor purity is critical. Although several methods have been proposed to predict tumor purity using high-throughput molecular data, there has been no comprehensive study on machine learning-based methods for the estimation of tumor purity. We applied various machine learning models to estimate tumor purity. Overall, the models predicted the tumor purity accurately and showed a high correlation with well-established gold standard methods. In addition, we identified a small group of genes and demonstrated that they could predict tumor purity well. Finally, we confirmed that these genes were mainly involved in the immune system. The machine learning models constructed for this study are available at https://github.com/BonilKoo/ML_purity.


Absorption of prednisolone in patients with Crohn's disease.
The absorption of prednisolone in patients with Crohn's disease was investigated. Seven patients with Crohn's disease and eight normal control subjects were given a tracer dose of tritiated prednisolone with 20 mg cold prednisolone by mouth. On a separate occasion they were given an intravenous injection of radiolabelled prednisolone. After oral ingestion only 53.4 +/- 11.7% of labelled material was excreted in the urine of Crohn's patients compared with 82.5 +/- 3.6% in the normal subjects. The oral/intravenous availability ratio was 0.61 +/- 0.14 in Crohn's patients and 0.89 +/- 0.07 in the normal group. Areas under plasma concentration-time curves were lower in patients than normal subjects and the oral/intravenous ratios were 0.6 +/- 0.2 and 0.86 +/- 0.09 respectively. Faecal excretion of radioactivity after oral ingestion was greater in Crohn's patients (19.3 +/- 2.5%, n = 3) than in normal subjects (7 +/- 2.8%, n = 4). The range for each type of measurement was much wider in the patient group than in the normal subjects. These data suggest that patients with Crohn's disease do not absorb prednisolone normally and that absorption varies between patients.


Novel N2-substituted pyrazolo[3,4-d]pyrimidine adenosine A3 receptor antagonists: inhibition of A3-mediated human glioblastoma cell proliferation.
Adenosine induces glioma cell proliferation by means of an antiapoptotic effect, which is blocked by cotreatment with selective A(3) AR antagonists. In this study, a novel series of N(2)-substituted pyrazolo[3,4-d]pyrimidines 2a-u was developed as highly potent and selective A(3) AR antagonists. The most performing compounds were derivatives 2a (R(1) = CH(3) and R(2) = COC(6)H(5); K(i) 334, 728, and 0.60 nM at the human A(1), A(2A), and A(3) ARs, respectively) and 2b (R(1) = CH(3) and R(2) = COC(6)H(4)-4-OCH(3); K(i) 1037, 3179, and 0.18 nM at the human A(1), A(2A), and A(3) ARs, respectively), which counteracted the effect of the A(3) AR agonists Cl-IB-MECA and IB-MECA on human glioma U87MG cell proliferation. This effect was concentration-dependent, with IC(50) values comparable to A(3) AR binding affinity values of 2a and 2b, thereby suggesting that their effects were receptor-mediated. Furthermore, the antiproliferative activity of the new compounds was demonstrated to be mediated by the block of A(3) AR agonist activation of intracellular kinases ERK 1/2.


Decoding alarm signal propagation of seed-harvester ants using automated movement tracking and supervised machine learning.
Alarm signal propagation through ant colonies provides an empirically tractable context for analysing information flow through a natural system, with useful insights for network dynamics in other social animals. Here, we develop a methodological approach to track alarm spread within a group of harvester ants, Pogonomyrmex californicus. We initially alarmed three ants and tracked subsequent signal transmission through the colony. Because there was no actual standing threat, the false alarm allowed us to assess amplification and adaptive damping of the collective alarm response. We trained a random forest regression model to quantify alarm behaviour of individual workers from multiple movement features. Our approach translates subjective categorical alarm scores into a reliable, continuous variable. We combined these assessments with automatically tracked proximity data to construct an alarm propagation network. This method enables analyses of spatio-temporal patterns in alarm signal propagation in a group of ants and provides an opportunity to integrate individual and collective alarm response. Using this system, alarm propagation can be manipulated and assessed to ask and answer a wide range of questions related to information and misinformation flow in social networks.


The CRM1 nuclear export protein in normal development and disease.
CRM1 (Chromosomal Maintenance 1, also known as Exportin 1) is the major mammalian export protein that facilitates the transport of large macromolecules including RNA and protein across the nuclear membrane to the cytoplasm. The gene encoding CRM1 was originally identified in yeast as required to maintain higher order chromosome structure. In mammalian cells, CRM1 was found to bind several nuclear pore proteins hence its role in nuclear-cytosolic transport was discovered. In addition to nuclear-cytosolic transport, CRM1 also plays a role in centrosome duplication and spindle assembly, especially in response to DNA damage. The crystal structure of CRM1 suggests a complex protein that binds the Ran protein bound to GTP, allowing for a conformational change that facilitates binding to different cargo proteins through a nuclear export signal (NES). Included in the cadre of cargo are multiple tumor suppressor and oncoproteins as p53, BRCA1, Survivin, NPM, and APC, which function in the nucleus to regulate transcription or aid in chromosomal assembly and movement. An imbalance in the cytosolic level of these proteins has been observed in cancer cells, resulting in either inactivation (tumor suppressor) or an excess of anti-apoptotic activity (oncoprotein). Thus, the concept of inhibiting CRM1 has been explored as a potential therapeutic intervention. Indeed, inhibition of CRM1 by a variety of small molecules that interfere with cargo-NES binding results in cancer cell death. Whether all of these proteins together are responsible for this phenotype or whether specific proteins are required for this effect is unclear at this time.


Evolutionary aspects of immunoglobulin heavy chain variable region (VH) gene subgroups.
We isolated and determined the sequences of two human germ-line heavy chain variable region (VH) genes and compared them with mouse VH genes. The results show that the human VHI subgroup is evolutionarily related to the mouse VHII subgroup. Evolutionary preservation of homologies in VH genes of the same subgroup includes not only the coding region but also intron size and homology in noncoding regions. This suggests that a VH gene subgroup constitutes a multigene family that undergoes concerted evolution. The homology between genes of the same subgroup in different species is greater than that between genes of different subgroups within a species. One of the VHII genes contains, in complementarity-determining region 2 (CDR2), a 13-base-pair previously shown to be in CDR2 of a VHIII gene and in a heavy chain diversity region gene, DH [Wu, T. T. & Kabat, E. A. (1982) Proc. Natl. Acad. Sci. USA 79, 5031-5032], suggesting the insertion of diversity region gene sequences into the VH gene. One of the human VH genes is a pseudogene because of a terminator, which, together with our previous results, shows that the VH gene repertoire contains 40% pseudogenes. In one of the VH genes, direct and inverted repeats at both 5' and 3' ends of the gene suggest a potential transposable element that encompasses the entire VH gene. It is possible that such a structure may facilitate saltatory replication and rapid expansion of VH gene families.


Long-term psychiatric care in Ontario: the Homes for Special Care Program.
During the last decade in Ontario large numbers of patients with chronic psychiatric disorders have been discharged from the mental hospitals and are now scattered throughout other psychiatric facilities. The Homes for Special Care Program offers privately run but government-funded accommodation for severely disabled patients with relatively stable and socially acceptable behaviour, who require residential or nursing care but are thought unlikely to benefit from further hospital treatment. Salient features of the program include the formal discharge of patients from hospital and their legal reinstatement as "persons", the cessation of active psychiatric treatment, and the provision of ongoing care and supervision by largely untrained personnel. Medical care is provided by general practitioners and the program looks to volunteer agencies to provide recreational and other activities for residents.


What aspects of others' behaviors do infants attend to in live situations?
A head-mounted camera was used for studying infant focus of attention. In two situations, 10- and 14-month-old infants observed two adults interacting. In one situation, the adults had a conversation and in the other situation, they were playing with blocks. The results indicate a preference for observing manual actions and a different pattern in looking at conversations than has been shown in eye-tracking studies. The head-mounted camera is a promising method for examining the infant's focus of attention.


Can poly-parameter linear-free energy relationships (pp-LFERs) improve modelling bioaccumulation in fish?
A wide range of studies have characterized different types of biosorbent, with regard to their interactions with chemicals. This has resulted in the development of poly-parameter linear free energy relationships (pp-LFERs) for the estimation of partitioning of neutral organic compounds to biological phases (e.g., storage lipids, phospholipids and serum albumins). The aims of this study were to explore and evaluate the influence of implementing pp-LFERs both into a one-compartment fish model and a multi-compartment physiologically based toxicokinetic (PBTK) fish model and the associated implications for chemical risk assessment. For this purpose, fish was used as reference biota, due to their important role in aquatic food chains and dietary exposure to humans. The bioconcentration factor (BCF) was utilized as the evaluation metric. Overall, our results indicated that models incorporating pp-LFERs (R2 = 0.75) slightly outperformed the single parameter (sp) LFERs approach in the one-compartmental fish model (R2 = 0.72). A pronounced enhancement was achieved for compounds with log KOW between 4 and 5 with increased R2 from 0.52 to 0.71. The minimal improvement was caused by the overestimation of lipid contribution and underestimation of protein contribution by the sp-approach, which cancelled each other out. Meanwhile, a greater improvement was observed for multi-compartmental PBTK models with consideration of metabolism, making all predictions fall within a factor of 10 compared with measured data. For screening purposes, the KOW-based (sp-LFERs) approach should be sufficient to quantify the main partitioning characteristics. Further developments are required for the consideration of ionization and more accurate quantification of biotransformation in biota.


Ultrastructural effects of cisplatin on the inner ear and lateral line system of zebrafish (Danio rerio) larvae.
Zebrafish, Danio rerio, has been a prominent model vertebrate for the study of chemical toxicity and human disease. Zebrafish hair cells (HCs) show significant structural, functional and molecular similarities to the mammalian inner ear HCs. We examined the effects of cisplatin, an anti-cancer drug, on HCs of the inner ear and on HCs and support cells (SCs) of neuromasts in zebrafish using transmission and scanning electron microscopy. Forty-five zebrafish larvae, 12 days post-fertilization, were assessed: 15 unexposed controls, 15 exposed to 10 µM cisplatin solution, and 15 exposed to 50 µM cisplatin solution. Hair cells in the cristae and maculae of the inner ear and of neuromasts were extremely sensitive to cisplatin. The drug was associated with vacuolization and the presence of myelinoid bodies in HC cytoplasm and with a condensation of the nuclear chromatin. The predominant pattern of injury was widespread degeneration of mitochondria, which appeared swollen and less electron-dense with disorganized or reduced cristae. Severity of damage seemed to be concentration-dependent, and the inner ear suffered more damage than the lateral line. Alterations similar to those in HCs were also observed in SCs of the neuromasts. Scanning electron microscopy showed loss of kinocilia in neuromasts of fish exposed to the higher concentration of cisplatin.


Understanding neurodevelopmental outcomes of prematurity: education priorities for NICU parents.
NICU nurses have an important role to play in many aspects of parent education. This article stresses the need for an increased focus on teaching parents about the central role that self-regulation will play in their infant's neurobehavioral development not only during the early infancy period but throughout all of childhood and adolescence. Suggestions are made about how to conceptualize the relation between cognition and emotions in humans and how to help parents understand that continued vigilance concerning potential problems in attention and self-regulation will be necessary.


Assessment of allergenicity of diploid and hexaploid wheat genotypes: identification of allergens in the albumin/globulin fraction.
Wheat is an important part of the daily diet of millions of people. However, this staple food is also responsible for food allergies. Ancient cultivars of wheat are gaining interest today but nothing is known about their allergenicity. Many wheat proteins have been reported as causative food allergens, including some prolamin-type gluten proteins, and salt soluble proteins of the albumin/globulin (A/G) type. The objective of this work is to obtain information about the allergenicity of the salt soluble A/G fraction of an ancient diploid cultivar compared with a standard hexaploid bread wheat cultivar using 20 sera from patients with wheat allergy. Differences in the IgE reactivity of sera towards the two genotypes were quantified by ELISA. Qualitative differences in IgE-binding proteins were searched after 1D or 2D electrophoresis. For most of the sera, the concentration in A/G specific IgE was higher for the hexaploid T. aestivum (cv Récital) than for the diploid T. monococcum (cv Engrain). The analysis of 2D spots revealed by immunoblotting leads to the identification by mass spectrometry of 39 IgE-binding proteins, some of them unknown until now as wheat allergens. Numerous allergens were identified, differences observed between Engrain and Récital will be discussed.


Regulation of particle morphology of pH-dependent poly(epsilon-caprolactone)-poly(gamma-glutamic acid) micellar nanoparticles to combat breast cancer cells.
The advantage of polymeric drug carriers lies in the uptake of the polymer nanoparticles by cancer cells before they release the drug, thereby reducing its toxic effects on healthy cells. A poly(gamma-glutamic acid)-b-poly(epsilon-caprolactone)-b-poly(gamma-glutamic acid) block copolymer was synthesized to encapsulate the anti-cancer drug doxorubicin in the treatment of wild type human breast cancer cells (MCF-7/WT). This pH-controllable carrier is negatively-charged in the presence of healthy tissues leading to lower cellular uptake. On the other hand, it becomes more hydrophobic in the acidic environment of cancer tissues, increasing its cellular uptake through the lipid bilayer. The block copolymer was characterized using Fourier transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, differential scanning calorimetry and dynamic light scattering. The micelles formed at a critical concentration range of 62-130 microg/mL depending on the composition of poly(gamma-glutamic acid) and poly(epsilon-caprolactone) chains. The nano-sized micelles were found to have pH-dependent sizes in the range of 90-200 nm. The role of poly(gamma-glutamic acid) was to increase the hydrophilicity and decrease the particle size of the copolymer. The structures of micelles that were more compact and less anionic showed better stability in plasma. It was found that the drug loading content and drug loading efficiency were 12.14% and 97.22% respectively. The copolymer showed shrinking and aggregation at low pH which led to a slower drug release. These nano-sized micelles showed potential as effective drug delivery carriers for doxorubicin because of its accumulation and slow release inside the MCF-7/WT cells.


Gut microbiota composition in depressive disorder: a systematic review, meta-analysis, and meta-regression.
Studies investigating gut microbiota composition in depressive disorder have yielded mixed results. The aim of our study was to compare gut microbiome between people with depressive disorder and healthy controls. We did a meta-analysis and meta-regression of studies by searching PubMed, Web of Science, Embase, Scopus, Ovid, Cochrane Library, ProQuest, and PsycINFO for articles published from database inception to March 07, 2022. Search strategies were then re-run on 12 March 2023 for an update. We undertook meta-analyses whenever values of alpha diversity and Firmicutes, Bacteroidetes (relative abundance) were available in two or more studies. A random-effects model with restricted maximum-likelihood estimator was used to synthesize the effect size (assessed by standardized mean difference [SMD]) across studies. We identified 44 studies representing 2091 patients and 2792 controls. Our study found that there were no significant differences in patients with depressive disorder on alpha diversity indices, Firmicutes and Bacteroidetes compared with healthy controls. In subgroup analyses with regional variations(east/west) as a predictor, patients who were in the West had a lower Chao1 level (SMD -0.42[-0.74 to -0.10]). Subgroup meta-analysis showed Firmicutes level was decreased in patients with depressive disorder who were medication-free (SMD -1.54[-2.36 to -0.72]), but Bacteroidetes level was increased (SMD -0.90[0.07 to 1.72]). In the meta-regression analysis, six variables cannot explain the 100% heterogeneity of the studies assessing by Chao1, Shannon index, Firmicutes, and Bacteroidetes. Depleted levels of Butyricicoccus, Coprococcus, Faecalibacterium, Fusicatenibacter, Romboutsia, and enriched levels of Eggerthella, Enterococcus, Flavonifractor, Holdemania, Streptococcus were consistently shared in depressive disorder. This systematic review and meta-analysis found that psychotropic medication and dietary habit may influence microbiota. There is reliable evidence for differences in the phylogenetic relationship in depressive disorder compared with controls, however, method of measurement and method of patient classification (symptom vs diagnosis based) may affect findings. Depressive disorder is characterized by an increase of pro-inflammatory bacteria, while anti-inflammatory butyrate-producing genera are depleted.


Application of physical principles in the development of tonometry.
A review of the development of applanation tonometry is undertaken from the perspective of the physical principles involved. Original articles that have contributed to this process are discussed. The result of this endeavour leads to a conclusion that corneal thinning procedures have no effect on original intraocular pressure. Also concluded is a methodology that would allow the direct measurement of original intraocular pressure.


Uterine rupture as a postpartum complication in two mares.
Uterine rupture was diagnosed in 2 postpartum mares with hemorrhagic vaginal discharge. Both mares had abdominal pain, as evidenced by pawing, kicking at the abdomen, or attempting to roll. Peritoneal fluid analysis was useful in establishing a diagnosis. One mare had many RBC in the peritoneal fluid and was anemic; this mare was managed medically with oxytocin, antibiotics, and blood transfusion. The mare was able to raise her foal to weaning age. The second mare had many RBC, degenerate neutrophils, and intracellular and extracellular bacteria in peritoneal fluid. Surgical repair of the uterus was performed, and the mare was treated for peritonitis. The mare later became pregnant.


Pure omega 3 polyunsaturated fatty acids (EPA, DPA or DHA) are associated with increased plasma levels of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) in a short-term study in women.
3-Carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF) is a metabolite of furan fatty acids found in plasma and urine of humans after consumption of foods containing these fatty acids. Recently, CMPF has been identified as a prominent metabolite following the consumption of either fish oil, fish oil fatty acid-ethyl esters or diets rich in fish. As furan fatty acids are known to occur in fish and fish oils (at a low level), it is possible that in these studies the CMPF in plasma originated from furan fatty acids. We report the plasma CMPF levels in 10 healthy women who consumed 1 gram of pure eicosapentaenoic acid (EPA), or docosapentaenoic acid (DPA) or docosahexaenoic acid (DHA), or olive oil daily for 6 days, in a cross-over study. The supplemented omega 3 fatty acids contained no detectable levels of furan fatty acids. The plasma CMPF and omega 3 fatty acid levels were measured by LC-MS/MS. Consumption of pure omega 3 fatty acids led to a significant increase in the plasma CMPF levels, but not with olive oil (from 1.6 to 2.5-fold compared with baseline). The plasma free fatty acid levels of EPA, DPA and DHA also increased significantly when they were supplemented (p < 0.05). Significant positive correlations existed between the plasma free fatty acid DPA and DHA levels (p < 0.05 and r = +0.49 to +0.81), but not between the EPA and CMPF levels. These data suggest that purified long chain omega 3 fatty acids may be precursors of CMPF; however the metabolic pathway(s) from omega 3 fatty acids to CMPF remain to be elucidated.


Co-inhibition of Aurora A and Haspin kinases enhances survivin blockage and p53 induction for mitotic catastrophe and apoptosis in human colorectal cancer.
Colorectal cancer (CRC) is a leading cause and mortality worldwide. Aurora A and haspin kinases act pivotal roles in mitotic progression. However, the blockage of Aurora A and Haspin for CRC therapy is still unclear. Here we show that the Haspin and p-H3T3 protein levels were highly expressed in CRC tumor tissues of clinical patients. Overexpression of Haspin increased the protein levels of p-H3T3 and survivin in human CRC cells; conversely, the protein levels of p-H3T3 and survivin were decreased by the Haspin gene knockdown. Moreover, the gene knockdown of Aurora A induced abnormal chromosome segregation, mitotic catastrophe, and cell growth inhibition. Combined targeted by co-treatment of CHR6494, a Haspin inhibitor, and MLN8237, an Aurora A inhibitor, enhanced apoptosis and CRC tumor inhibition. MLN8237 and CHR6494 induced abnormal chromosome segregation and mitotic catastrophe. Meanwhile, MLN8237 and CHR6494 inhibited survivin protein levels but conversely induced p53 protein expression. Ectopic survivin expression by transfection with a survivin-expressed vector resisted the cell death in the MLN8237- and CHR6494-treated cells. In contrast, the existence of functional p53 increased the apoptotic levels by treatment with MLN8237 and CHR6494. Co-treatment of CHR6494 and MLN8237 enhanced the blockage of human CRC xenograft tumors in nude mice. Taken together, co-inhibition of Aurora A and Haspin enhances survivin inhibition, p53 pathway induction, mitotic catastrophe, apoptosis and tumor inhibition that may provide a potential strategy for CRC therapy.


Diffusion tensor imaging of the spinal cord and its clinical applications.
The identification of the extent of neural damage in patients with acute or chronic spinal cord injury is imperative for the accurate prediction of neurological recovery. The changes in signal intensity shown on routine MRI sequences are of limited value for predicting functional outcome. Diffusion tensor imaging (DTI) is a novel radiological imaging technique which has the potential to identify intact nerve fibre tracts, and has been used to image the brain for a variety of conditions. DTI imaging of the spinal cord is currently only a research tool, but preliminary studies have shown that it holds considerable promise in predicting the severity of spinal cord injury. This paper briefly reviews our current knowledge of this technique.


Functional microbial diversity explains groundwater chemistry in a pristine aquifer.
The diverse microbial populations that inhabit pristine aquifers are known to catalyze critical in situ biogeochemical reactions, yet little is known about how the structure and diversity of this subsurface community correlates with and impacts upon groundwater chemistry. Herein we examine 8,786 bacterial and 8,166 archaeal 16S rRNA gene sequences from an array of monitoring wells in the Mahomet aquifer of east-central Illinois. Using multivariate statistical analyses we provide a comparative analysis of the relationship between groundwater chemistry and the microbial communities attached to aquifer sediment along with those suspended in groundwater. Statistical analyses of 16S rRNA gene sequences showed a clear distinction between attached and suspended communities; with iron-reducing bacteria far more abundant in attached samples than suspended, while archaeal clones related to groups associated with anaerobic methane oxidation and deep subsurface gold mines (ANME-2D and SAGMEG-1, respectively) distinguished the suspended community from the attached. Within the attached bacterial community, cloned sequences most closely related to the sulfate-reducing Desulfobacter and Desulfobulbus genera represented 20% of the bacterial community in wells where the concentration of sulfate in groundwater was high (> 0.2 mM), compared to only 3% in wells with less sulfate. Sequences related to the genus Geobacter, a genus containing ferric-iron reducers, were of nearly equal abundance (15%) to the sulfate reducers under high sulfate conditions, however their relative abundance increased to 34% when sulfate concentrations were < 0.03 mM. Also, in areas where sulfate concentrations were <0.03 mM, archaeal 16S rRNA gene sequences similar to those found in methanogens such as Methanosarcina and Methanosaeta comprised 73-80% of the community, and dissolved CH4 ranged between 220 and 1240 μM in these groundwaters. In contrast, methanogens (and their product, CH4) were nearly absent in samples collected from groundwater samples with > 0.2 mM sulfate. In the suspended fraction of wells where the concentration of sulfate was between 0.03 and 0.2 mM, the archaeal community was dominated by sequences most closely related to the ANME-2D, a group of archaea known for anaerobically oxidizing methane. Based on available energy (∆GA) estimations, results varied little for both sulfate reduction and methanogenesis throughout all wells studied, but could favor anaerobic oxidation of methane (AOM) in wells containing minimal sulfate and dihydrogen, suggesting AOM coupled with H2-oxidizing organisms such as sulfate or iron reducers could be an important pathway occurring in the Mahomet aquifer. Overall, the results show several distinct factors control the composition of microbial communities in the Mahomet aquifer. Bacteria that respire insoluble substrates such as iron oxides, i.e. Geobacter, comprise a greater abundance of the attached community than the suspended regardless of groundwater chemistry. Differences in community structure driven by the concentration of sulfate point to a clear link between the availability of substrate and the abundance of certain functional groups, particularly iron reducers, sulfate reducers, methanogens, and methanotrophs. Integrating both geochemical and microbiological observations suggest that the relationships between these functional groups could be driven in part by mutualism, especially between ferric-iron and sulfate reducers.


Meta-analysis of antiarrhythmic drug trials.
During the past 15 years, the efficacy of antiarrhythmic drugs has been investigated for reducing premature death in patients at high risk of arrhythmia. Whereas the benefits of beta-blocker therapy are well established, a reduction in mortality with other antiarrhythmic drugs remains unproved and in some cases, there is evidence of increased mortality with class I and some class III agents. A limitation of individual clinical trials is inadequate sample size to detect significant differences between interventions. Meta-analysis, by combining results from multiple clinical trials, provides a technique to overcome sample size limitations and assess the benefits and limitations of an intervention. Thirteen randomized clinical trials evaluated the role of prophylactic amiodarone in patients at risk of death from cardiac arrhythmias. Whereas 3 of these studies reported a reduction in mortality, several others revealed no benefits of amiodarone. Because neither trial was designed to detect reductions in total mortality, it remained unclear whether the beneficial effect of amiodarone on arrhythmic death and resuscitated ventricular fibrillation translated into a beneficial effect on total mortality. To address this, a meta-analysis was performed from the 13 trials of amiodarone in patients after an acute myocardial infarction or with congestive heart failure. The results showed a significant reduction in mortality and in arrhythmic death with amiodarone.


Expansion of human articular chondrocytes and formation of tissue-engineered cartilage: a step towards exploring a potential use of matrix-induced cell therapy.
Monolayer culture expansion remains as a fundamental step to acquire sufficient number of cells for 3D constructs formation. It has been well-documented that cell expansion is however accompanied by cellular dedifferentiation. In order to promote cell growth and circumvent cellular dedifferentiation, we evaluated the effects of Transforming Growth Factor Beta-2 (TGF-β2), Insulin-like Growth Factor-I (IGF-I) and basic Fibroblast Growth Factor (bFGF) combination on articular chondrocytes culture and 'chondrocytes-fibrin' construct formation. Chondrocytes were serially cultured in: (1) F12:DMEM+10% Foetal Bovine Serum (FBS) with growth factors (FD10GFs), (2) F12:DMEM+2%FBS with the growth factors (FD2GFs) and, (3) F12:DMEM+10%FBS without growth factors (FD) as control. Cultured chondrocytes were evaluated by means of growth kinetics parameters, cell cycle analysis, quantitative phenotypic expression of collagen type II, aggrecan core protein sox-9 and collagen type I and, immunochemistry technique. Harvested chondrocytes were incorporated with plasma-derived fibrin and were polymerized to form the 3D constructs and implanted subcutaneously at the dorsum of athymic nude mice for eight (8) weeks. Resulted constructs were assigned for gross inspections and microscopic evaluation using standard histochemicals staining, immunochemistry technique and, quantitative phenotypic expression of cartilage markers to reassure cartilaginous tissue formation. Growth kinetics performance of chondrocytes cultured in three (3) types of culture media from the most to least was in the following order: FD10GFs>FD2GFs>FD. Following growth kinetics analysis, we decided to use FD10GFs and FD (control) for further evaluation and 'chondrocytes-fibrin' constructs formation. Chondrocytes cultured in FD10GFs preserved the normal diploid state (2c) with no evidence of aneuploidy, haploidy or tetraploidy. Expression of cartilage-specific markers namely collagen type II, aggrecan core protein and sox-9 were significantly higher in FD10GFs when compared to control. After implantation, 'chondrocytes-fibrin' constructs exhibited firm, white, smooth and glistening cartilage-like properties. FD10GFs constructs formed better quality cartilage-like tissue than FD constructs in term of overall cartilaginous tissue formation, cells organization and extracellular matrix distribution in the specimens. Cartilaginous tissue formation was confirmed by the presence of lacunae and cartilage-isolated cells embedded within basophilic ground substance. Presence of proteoglycan was confirmed by positive Safranin O staining. Collagen type II exhibited immunopositivity at the pericellular and inter-territorial matrix area. Chondrogenic properties of the construct were further confirmed by the expression of genes encoding collagen type II, aggrecan core protein and sox9. In conclusion, FD10GFs promotes the proliferation of chondrocytes and formation of good quality 'chondrocytes-fibrin' constructs which may have potential use of matrix-induced cell implantation.


Use of technology to increase access to mental health services in a rural Texas community.
The Leon County Health Resource Commission sought to increase access to mental health services for their rural community. The commission formed a network of partners who collaborated to increase free transportation to mental health services outside the community and developed a telehealth-based counseling program through a counseling psychology training program. Learning opportunities emerged during the development and implementation of these activities for both the students and the community in how to successfully utilize and sustain this service. This article describes the telehealth counseling model, presents lessons learned in the process, and presents recommendations for others interested in utilizing similar strategies.


Vernalization treatment induces site-specific DNA hypermethylation at the VERNALIZATION-A1 (VRN-A1) locus in hexaploid winter wheat.
Certain temperate species require prolonged exposure to low temperature to initiate transition from vegetative growth to flowering, a process known as vernalization. In wheat, winter cultivars require vernalization to initiate flowering, making vernalization requirement a trait of key importance in wheat agronomy. The genetic bases of vernalization response have been largely studied in wheat, leading to the characterization of a regulation pathway that involves the key gene VERNALIZATION1 (VRN1). While previous studies in wheat and barley have revealed the functional role of histone modification in setting VRN1 expression, other mechanisms might also be involved. Here, we were interested in determining whether the cold-induced expression of the wheat VRN-A1 gene is associated with a change in DNA methylation. We provide the first DNA methylation analysis of the VRN-A1 gene, and describe the existence of methylation at CG but also at non CG sites. While CG sites show a bell-shape profile typical of gene-body methylation, non CG methylation is restricted to the large (8.5 kb) intron 1, in a region harboring fragments of transposable elements (TEs). Interestingly, cold induces a site-specific hypermethylation at these non CG sites. This increase in DNA methylation is transmitted through mitosis, and is reset to its original level after sexual reproduction. These results demonstrate that VRN-A1 has a particular DNA methylation pattern, exhibiting rapid shift within the life cycle of a winter wheat plant following exposure to particular environmental conditions. The finding that this shift occurs at non CG sites in a TE-rich region opens interesting questions onto the possible consequences of this type of methylation in gene expression.


A systematic review of interventions for treatment resistant major depressive disorder in adolescents.
To update a comparative effectiveness review (1980-2011) of treatments for adolescents whose depressive episode or disorder (MDE/MDD) did not respond to one or more trials of SSRI antidepressants. MEDLINE, Cochrane Central, PsychINFO, Cochrane Database of Systematic Reviews, EMBASE, CINAHL, and AMED were searched in addition to the grey literature. We spanned May 2011 to September 1, 2017 and included only articles in English. 11 new studies were reviewed based on the criteria of having tested a comparative treatment in adolescents with MDD or MDE who were confirmed to have failed one or more SSRI trials. Data were extracted using standardized forms and a reference guide in DistillerSR; a second reviewer verified the accuracy of the data fields and discrepancies were resolved by consensus. One trial (N = 29) found a small benefit of escalating doses of fluoxetine and the treatment of adolescent depression study (TORDIA, N = 334) found significant benefits of combined SSRI or venlafaxine treatment with CBT for most outcomes. No new studies were identified since the previous review (2012). One trial is currently registered that will be a cross over trial of rTMS; other registered trials are open label. Multiple secondary data analyses of TORDIA have identified important predictors of treatment response and relapse. No new comparative studies were identified since the original review. Trials are desperately needed to identify new treatments for youth with SSRI resistant MDD. These youth should not be deemed as treatment resistant until completing one or two failed trials of SSRI combined with evidence-based psychotherapy.


25-Hydroxyvitamin D response to cholecalciferol supplementation in hemodialysis.
Recent understanding of extrarenal production of calcitriol has led to the exploration of native vitamin D treatment in dialysis patients. This paper reports the pharmacokinetics of 25-hydroxyvitamin D response to 10,333 IU cholecalciferol given weekly in subjects on chronic dialysis. This randomized, double-blind, placebo-controlled trial of 15 weeks of oral cholecalciferol in subjects with stage 5 CKD requiring maintenance hemodialysis was conducted from November of 2007 to March of 2010. The time course of serum 25-hydroxyvitamin D was measured over the course of treatment. Additionally, blood was drawn at baseline and last visit for calcium, phosphorus, calcitriol, and parathyroid hormone levels. The median (interquartile range) baseline 25-hydroxyvitamin D level was 13.3 (11.1-16.2) ng/ml for the treatment group and 15.2 (10.7-19.9) ng/ml for the placebo group. 25-hydroxyvitamin D steady state levels rose by 23.6 (19.2-29.9) ng/ml in the treatment group, and there was no change in the placebo group. Calcitriol levels also increased significantly in the treatment group. There were no significant changes in levels of calcium, albumin, phosphorus, and parathyroid hormone in either group. Cholecalciferol (10,333 IU) given weekly in patients on chronic hemodialysis produces a steady state in 25-hydroxyvitamin D of approximately 24 ng/ml.


Transmission of HIV infection to heterosexual partners but not to household contacts of seropositive haemophiliacs.
The prevalence of HIV antibodies in 44 heterosexual female partners and 56 nonsexual family household contacts of 61 HIV seropositive haemophiliacs (41 adults and 20 children or adolescents) was determined to evaluate the risk of transmission of HIV infection. HIV antibodies were determined by enzyme-linked immunosorbent assay and positive reactions were confirmed by Western blotting. HIV antibodies were demonstrated in 4/40 (10%) regular heterosexual partners of 40 seropositive patients with haemophilia A. Four temporary heterosexual partners of one additional seropositive haemophiliac were seronegative. 56 nonsexual household contacts including 30 parents, 13 siblings and 13 children of 29 seropositive haemophiliacs were all negative for HIV antibodies. Thus transmission of HIV occurred from seropositive haemophiliacs to their heterosexual partners but not to nonsexual household contacts.


[The ultrastructure of the natural killer cells in transitional-cell bladder cancer].
Electronic microexamination of two of the 65 cases of transitional cell carcinoma, in which cases we succeeded in detecting and studying the interaction between the natural killer cells and a target cell, suggested that a natural killer cell might be an active element of tissue immunity being capable of recognizing and destroying cells that differ in their genetic apparatus from normal cells and bodily tissues. The main targets of natural killer cells are those organelles in tumour cells that are their most active units of vital importance, the destruction of which is aimed at maintaining of tissue and cellular homeostasis.


Performance challenges for children and adolescents with difficulty processing and integrating sensory information: a systematic review.
A systematic review of the literature related to performance difficulties for children and adolescents with difficulty processing and integrating sensory information was completed as part of the Evidence-Based Literature Review Project of the American Occupational Therapy Association. The review focused on functional performance difficulties that these children may exhibit in areas of occupation including play and leisure, social participation, activities of daily living, instrumental activities of daily living, rest and sleep, education, and work. The results suggest that children and adolescents with difficulty processing and integrating sensory information do exhibit functional performance difficulties in key areas of occupation. However, further descriptive studies are needed to tie these difficulties to their specific sensory and motor issues. Researchers are encouraged to include functional performance measures and measures of social participation in their studies to further elucidate these relationships.


Microdeposition of amyloid in the joints.
The distribution of amyloid in various organs, including the hip and the sternoclavicular joints, was systemically investigated in a prospective necropsy study of 91 unselected individuals whose median age was 70 years. The overall incidence of amyloid microdeposits in both joints was 56%. Their presence correlated significantly with age only. The deposition began earlier in the sternoclavicular joint, where the fibrocartilaginous disc was almost exclusively affected. The myocardium was the next most frequently affected organ, with an incidence of 12.2%. Minor amyloidosis of the articular tissues had no significant clinical counterpart, neither was it associated with any particular disease. It seemingly represents a localised and isolated degenerative phenomenon, whose morphological aspects invite speculation about the pathogenic role of long-lasting mechanical stress.


In planta particle bombardment (iPB): A new method for plant transformation and genome editing.
Transformation is a key step in modern breeding technology that involves genome editing. The requirement for in vitro tissue culture and regeneration hampers application of this technology to commercially important varieties of many crop species. To overcome this problem, we developed a simple and reproducible in planta transformation method in wheat (Tritticum aestivum L.). Our in planta particle bombardment (iPB) method utilizes the shoot apical meristem (SAM) as a target tissue. The SAM contains a subepidermal cell layer termed L2, from which germ cells later develop during floral organogenesis. The iPB method can also be used for genome editing through transient CRISPR/Cas9 expression or direct delivery of the CRISPR/Cas9 ribonucleoprotein. In this review, we describe the iPB technology and provide an overview of its current and future applications in plant transformation and genome editing.


Sub-100-fs Kerr lens mode-locked Yb:Lu2O3 thin-disk laser oscillator operating at 21 W average power.
We investigate power-scaling of a Kerr lens mode-locked (KLM) Yb:Lu2O3 thin-disk laser (TDL) oscillator operating in the sub-100-fs pulse duration regime. Employing a scheme with higher round-trip gain by increasing the number of passes through the thin-disk gain element, we increase the average power by a factor of two and the optical-to-optical efficiency by a factor of almost three compared to our previous sub-100-fs mode-locking results. The oscillator generates pulses with a duration of 95 fs at 21.1 W average power and 47.9 MHz repetition rate. We discuss the cavity design for continuous-wave and mode-locked operation and the estimation of the focal length of the Kerr lens. Unlike to usual KLM TDL oscillators, an operation at the edge of the stability zone in continuous-wave operation is not required. This work shows that KLM TDL oscillators based on the gain material Yb:Lu2O3 are an excellent choice for power-scaling of laser oscillators in the sub-100-fs regime, and we expect that such lasers will soon operate at power levels in excess of hundred watts.


Fringe is a glycosyltransferase that modifies Notch.
Notch receptors function in highly conserved intercellular signalling pathways that direct cell-fate decisions, proliferation and apoptosis in metazoans. Fringe proteins can positively and negatively modulate the ability of Notch ligands to activate the Notch receptor. Here we establish the biochemical mechanism of Fringe action. Drosophila and mammalian Fringe proteins possess a fucose-specific beta1,3 N-acetylglucosaminyltransferase activity that initiates elongation of O-linked fucose residues attached to epidermal growth factor-like sequence repeats of Notch. We obtained biological evidence that Fringe-dependent elongation of O-linked fucose on Notch modulates Notch signalling by using co-culture assays in mammalian cells and by expression of an enzymatically inactive Fringe mutant in Drosophila. The post-translational modification of Notch by Fringe represents a striking example of modulation of a signalling event by differential receptor glycosylation and identifies a mechanism that is likely to be relevant to other signalling pathways.


Molecular cloning of membrane cofactor protein (MCP; CD46) on B95a cell, an Epstein-Barr virus-transformed marmoset B cell line: B95a-MCP is susceptible to infection by the CAM, but not the Nagahata strain of the measles virus.
Measles virus (MV) infects not only human beings but also some simian species. The MV receptor on Vero cells (a cell line established from African Green monkey kidney cells) and human cells has been shown to be the membrane cofactor protein MCP/CD46, which is an inhibitor of autologous complement (C) activation. B95a, an Epstein-Barr virus (EBV)-transformed marmoset B cell line, is a simian cell line used for MV selection and is much more susceptible to MV than Vero cells. In the present study, we isolated cDNAs encoding MCP homologues from B95a cDNA library and assessed whether B95a-MCP is responsible for the high susceptibility of B95a to MV. The deduced amino acid sequence of the cDNA of B95a-MCP was 76% identical to that of human-MCP, and the recombinant B95a-MCP exerts C inhibitor activity. Although CAM, a vaccine strain of MV, infected Chinese hamster ovary (CHO) cells expressing B95a-MCP, Nagahata strain, a wild type of MV, failed to infect the CHO transfectants, suggesting that additional membrane molecules of B95a are responsible for the high susceptibility of B95a to the Nagahata strain.


Snail transcription factors as key regulators of chemoresistance, stemness and metastasis of ovarian cancer cells.
Ovarian cancer is one of the deadliest gynecological malignancies among women. The reason for this outcome is the frequent acquisition of cancer cell resistance to platinum-based drugs and unresponsiveness to standard therapy. It has been increasingly recognized that the ability of ovarian cancer cells to adopt more aggressive behavior (mainly through the epithelial-to-mesenchymal transition, EMT), as well as dedifferentiation into cancer stem cells, significantly affects drug resistance acquisition. Transcription factors in the Snail family have been implicated in ovarian cancer chemoresistance and metastasis. In this article, we summarize published data that reveal Snail proteins not only as key inducers of the EMT in ovarian cancer but also as crucial links between the acquisition of ovarian cancer stem properties and spheroid formation. These Snail-related characteristics significantly affect the ovarian cancer cell response to treatment and are related to the acquisition of chemoresistance.


The influence of insulin on the metabolism of glucosamine in the isolated rat diaphragm muscle.
1. The influence of insulin on the metabolism of [1-(14)C]glucosamine by diaphragm muscle from normal rats and rats rendered diabetic with streptozotocin has been studied. 2. The glucosamine was converted into glucosamine 1-phosphate, glucosamine 6-phosphate, glycogen, lactate and small amounts of other unidentified intermediates. 3. Insulin increased the incorporation of (14)C into glycogen in both the normal and diabetic muscle, but did not increase the formation of the glucosamine phosphate esters. 4. The (14)C content in the glycogen was present partly as glucose and partly as glucosamine; there was significantly more [(14)C]glucose in the glycogen of the diabetic muscle than in that of the normal muscle.


Fluorescent and Catalytic Properties of a 2D Lamellar Zn Metal-Organic Framework with sql Network Structure.
A two-dimensional (2D) lamellar Zn metal-organic framework (Zn-MOF, 1) with a fluorescent 1,6-di(pyridin-3-yl)pyrene (3-DPPy) and 1,4-benzenedicarboxylate (BDC2-) bridging linkers was prepared and structurally characterized. The chemical formula of 1 is [Zn(μ-3-DPPy)(μ-BDC)]n. The mononuclear Zn(II) ion, acting as a node, is tetrahedrally coordinated with two 3-DPPy and two BDC linkers. The coordination environment of Zn(II) is a distorted tetrahedral geometry. The Zn-MOF is the sql network structure based on topology analysis. The undulated 2D sheets of 1 tightly pack together to form a lamellar structure. The pyrene moieties are parallelly oriented to each other. The Zn-MOF is not porous, possibly because the mononuclear Zn(II) node did not form cluster-based secondary building units due to the less symmetric 3-DPPy. The steady-state fluorescence measurements indicate that the fluorescence signal of the 1 is slightly blue-shifted compared to the free 3-DPPy in the solid state. The excimer emission band at 463 nm for crystalline 3-DPPy is shifted to 447 nm for 1. The value of 447 nm is also a blue-shift value compared to nonsubstituted pyrene crystals (470 nm). Despite its nonporosity, the surface Lewis acidic sites of 1 could catalyze the transesterification of esters. Surface defect sites are responsible for this catalytic activity.


Nanoscale Electronic Inhomogeneity in In2Se3 Nanoribbons Revealed by Microwave Impedance Microscopy.
Driven by interactions due to the charge, spin, orbital, and lattice degrees of freedom, nanoscale inhomogeneity has emerged as a new theme for materials with novel properties near multiphase boundaries. As vividly demonstrated in complex metal oxides (see refs 1-5) and chalcogenides (see refs 6 and 7), these microscopic phases are of great scientific and technological importance for research in high-temperature superconductors (see refs 1 and 2), colossal magnetoresistance effect (see ref 4), phase-change memories (see refs 5 and 6), and domain switching operations (see refs 7-9). Direct imaging on dielectric properties of these local phases, however, presents a big challenge for existing scanning probe techniques. Here, we report the observation of electronic inhomogeneity in indium selenide (In(2)Se(3)) nanoribbons (see ref 10) by near-field scanning microwave impedance microscopy (see refs 11-13). Multiple phases with local resistivity spanning 6 orders of magnitude are identified as the coexistence of superlattice, simple hexagonal lattice and amorphous structures with approximately 100 nm inhomogeneous length scale, consistent with high-resolution transmission electron microscope studies. The atomic-force-microscope-compatible microwave probe is able to perform a quantitative subsurface electrical study in a noninvasive manner. Finally, the phase change memory function in In(2)Se(3) nanoribbon devices can be locally recorded with big signals of opposite signs.


Hemangiopericytoma of the tracheal wall.
A hemangiopericytoma occurred on the posterior tracheal wall in a 46-year-old man. A review of the pathologic findings showed that compressed groups of capillaries were lined by an intact reticulin sheath that was surrounded by spindle-shaped pericytes. The ultramicroscopic findings of small spindle cells with a surrounding basal lamina and intracellular microfilaments in the absence of neurosecretory granules are important diagnostic features. Emphasis is placed on the impossibility of differentiating benign and malignant lesions pathologically, the need for long-term clinical follow-up, and the necessity for adequate complete surgical resection of these lesions. The use of a surgical approach to the posterior side of the tracheal wall can prevent postoperative stenosis.


[Modulation of S-1 conformation and inhibition of the skeletal muscle S-1-ATPase by calponin of the mussel].
A novel 40 kDa protein has been detected in native thin filaments from catch muscles of the mussel Crenomytilus grayanus. In this study, using skeletal muscle actin and S-1, we investigated the effects of the mussel 40-kDa actin-binding protein on the acto · S-1 ATPase activity. On increasing the 40-kDa actin-binding protein (CaP-40) concentration, the actin-activated ATPase activity decreased, and was inhibited 80% at a CaP-40 to actin ratio of 0.5. Polarized fluorimetry technique and glycerinated muscle fibers were used to study effects of CaP-40 on the orientation and mobility of fluorescent label 1.5-IAEDANS specifically bound to CyS-707 of myosin subfragment-1 in the absence of nucleotide, and in the presence of MgADP or MgATP. We have concluded that CaP-40 binding to actin affects the strong binding of myosin to actin but has no effect on the weak binding. Thus, the influence of the CaP-40 on the formation of strong actomyosin binding forms A · M and A · M · ADP manifests itself by a decrease in the relative content of myosin cross-bridges strongly bound with actin, which probably results in a decrease in the relative content of "switch on" actin monomers in thin filaments. This suggests that, as calponin CaP-40 selects its target the phase of strong actomyosin binding binding which preceded by a phase generating power stroke.


Singing on the wings! Male wing fanning performances affect female willingness to copulate in the aphid parasitoid Lysiphlebus testaceipes (Hymenoptera: Braconidae: Aphidiinae).
Lysiphlebus testaceipes (Hymenoptera: Braconidae: Aphidiinae) is a generalist endoparasitoid attacking more than 100 aphid species. In L. testaceipes, wing fanning is a main male courtship display evoked by a female-borne sex pheromone. However, no information is available on the characteristics and behavioral role of male fanning during courtship in this parasitoid. Here, the courtship behavior of a wild strain of L. testaceipes was quantified and the male wing fanning performances were analyzed through high-speed video recordings and examined in relation to mating success. Courtship sequence of wild L. testaceipes did not substantially differ from that previously reported for other populations mass reared on aphids. We observed that the male courtship duration did not affect mating success. However, video analysis revealed that the males producing high-frequency fanning signals achieved higher mating success over those that display low-frequency fanning. Wing fanning before successful and unsuccessful courtship differed in amplitude of wing movements and alignment toward the mate, highlighting that frontal courtship positively influence the female mating decisions. This study increases knowledge on sexual behavior in a key parasitoid of aphids, highlighting the importance of wing fanning among the range of sensory modalities used in the sexual communication of L. testaceipes. From a practical point of view, this information is useful in L. testaceipes-based biocontrol strategies, since it can help to establish parameters for quality checking of mass-reared wasps over time.


Expression pattern of BXR suggests a role for benzoate ligand-mediated signalling in hatching gland function.
The Xenopus laevis nuclear receptor BXR has recently been shown to be activated by a class of endogenous benzoate metabolites, indicating the presence of a novel and unsuspected benzoate ligand-dependent signalling pathway. The receptor is expressed ubiquitously in blastula and gastrula stage embryos, and its expression declines during neurula stages. In order to examine further this novel vertebrate signalling system, we have examined the expression of the BXR gene in tailbud stage embryos and adults. We show here that in Xenopus tailbud stage embryos expression is restricted to the hatching gland, suggesting a role in hatching gland function. Neither BXR nor a BXR-VP16 fusion is sufficient to specify hatching gland in neurally-induced tissue. In adults, BXR expression is abundant in the brain and gonads. This expression pattern in adults is distinct from any of the putative mammalian homologues. A nuclear receptor that mediates benzoate signalling has yet to be found in mammals.


Changes in Muscle Activity Patterns and Joint Kinematics During Gait in Hemophilic Arthropathy.
Hemophilic arthropathy is the result of repetitive intra-articular bleeding and synovial inflammation. In people with hemophilic arthropathy (PWHA), very little is known about the neural control of individual muscles during movement. The aim of the present study was to assess if the neural control of individual muscles and coordination between antagonistic muscle pairs and joint kinematics during gait are affected in PWHA. Thirteen control subjects (CG) walked overground at their preferred and slow velocity (1 m/s), and 14 PWHA walked overground at the preferred velocity (1 m/s). Joint kinematics and temporal gait parameters were assessed using four inertial sensors. Surface electromyography (EMG) was collected from gluteus maximus (GMAX), gluteus medius (GMED), vastus medialis (VM), vastus lateralis (VL), rectus femoris (RF), medial gastrocnemius (MG), lateral gastrocnemius (LG), soleus (SOL), tibialis anterior (TA), semitendinosus (ST), and biceps femoris (BF). Waveforms were compared using the time-series analysis through statistical parametric mapping. In PWHA compared to CG, EMG amplitude during the stance phase was higher for LG (for both velocities of the CG), BF (slow velocity only), and ST (preferred velocity only) (p < 0.05). Co-contraction during the stance phase was higher for MG-TA, LG-TA, VL-BF, VM-ST, LG-VL, and MG-VM (both velocities) (p < 0.05). MG and LG were excited earlier (preferred velocity only) (p < 0.05). A later offset during the stance phase was found for VL, BF, and ST (both velocities), and BF and GMAX (preferred velocity only) (p < 0.05). In addition, the range of motion in knee and ankle joints was lower in PWHA (both velocities) and hip joint (preferred velocity only) (p < 0.05). In conclusion, the neural control of individual muscles and coordination between antagonistic muscles during gait in PWHA differs substantially from control subjects.


Diagnosing respiratory events and tracing air flow by internal thermistors.
We have developed a new method to measure flow in patients with sleep-related breathing disorders (SRBD). These flow sensors are modified thermistors located in the same sensors we use for pressure measurement in the upper airways to find the obstructive segments during apnoeas. The aim of this study was to test if using internal thermistors as indicator of air flow has advantages compared with the external thermistor method in detecting respiratory events. A total of 50 consecutive patients with an apnoea-hypopnoea index (AHI) of more than 15 were studied. A standard nocturnal polysomnography (PSG) with both internal and external thermistors was performed in all patients. To estimate the patients' AHI, a detailed analysis viewing all parameters except external and internal thermistors was performed. This was followed by an analysis viewing only internal thermistors signals indicating airflow in the pharynx, and finally an analysis viewing only external thermistor signals indicating air flow at the mouth and nose. Mean AHI measured by the three methods showed 49.8 (SD 23.4) by the PSG, 47.8 (SD 24.9) by internal thermistors alone, and 31.5 (SD 22.2) by external thermistors. There was no statistical difference between AHI detected by PSG and internal thermistors, but highly significant differences between PSG and external thermistors (p < 0.001). The external thermistors missed an average of almost 20 respiratory events per hour.


Detection of Uveal Melanoma by Multiplex Immunoassays of Serum Biomarkers.
Multiplex immunoassays simultaneously measure multiple analytes in a single sample providing quantitative data via parallel analyses, which is especially suitable for serum biomarker verification and validation. Multiplex immunoassays demonstrate several advantages over traditional enzyme-linked immunosorbent assays such as increasing productivity, conserving critical reagents and samples, and delivering results quickly. Here we describe the detection of uveal melanoma by magnetic bead-based multiplex immunoassays of serum biomarkers. The biomarker panels evaluated by multiplex immunoassays with high analytical performance demonstrated potential complementary values in detection of uveal melanoma.


Pattern of alcohol consumption and associated factors among adolescents students of public schools in an inner city in Brazil.
To identify issues related to alcohol consumption among students from an inner city in Southeastern Brazil. In this cross-sectional study, 638 students from 13 to 17 years old, of which 355 (55.6%) were girls, were randomly selected in 13/40 (32.5%) public schools from November 2009 to August 2010. A structured questionnaire was used to collect personal/family data, and the Alcohol Use Disorders Identification Test (AUDIT) was used to evaluate alcohol consumption. Bivariate analysis and logistic regression were used. It was observed that 516 (80.9%) students have used alcohol in their lifetime, and 115 (18.4%) had hazardous drinking (AUDIT ≥ 8). By crude analysis, the hazardous drinking was associated with age range (16 - 17 years old), age at first alcohol consumption (≤ 12 years old), higher family income, conflicting family environment, absence of religion practice and parents that consume alcoholic beverages. By adjusted analysis, it was observed an association with the age at first alcohol consumption [≤ 12 years old, odds ratio (OR) = 2.5; 95% confidence interval (95%CI) 1.4 - 4.4]. Active sex life was more frequent among those that have used alcohol in their lifetime (OR = 3.3; 95%CI 2.0 - 5.3). Under the influence of alcohol, 22/103 (21.4%) students have not always used condoms. Among all, 25.4% believe that there is no risk in drinking, and 98% had already bought alcoholic beverages. Having the desire to drink after watching alcoholic beverage advertisements was more frequent among adolescents who had already consumed these beverages (OR = 1.7; 95%CI 1.1 - 2.6). The alcohol consumption is early and worrying among adolescents, emphasizing the need for awareness of students and their parents and carers to these risks, and also the compliance with the law prohibiting the sale of alcoholic beverages to minors.


High incidence rate of vertebral fractures during chronic prednisone treatment, in spite of bisphosphonate or alfacalcidol use. Extension of the alendronate or alfacalcidol in glucocorticoid-induced osteoporosis-trial.
In the 18 month "alendronate or alfacalcidol in glucocorticoid-induced osteoporosis"-trial (STOP-trial) patients with rheumatic diseases who started glucocorticoids were randomised to anti-osteoporosis therapy with either daily alendronate (10 mg) or alfacalcidol (1 microg). In the present observational open follow-up study of the STOP-trial, we report the long-term effects of risk factors on the incidence and pattern of vertebral fractures, assessed using the Genant method. Of the 201 included patients in the STOP-trial, 163 completed the trial and of those 116 underwent a follow-up radiography of the spine. Twenty-eight patients had developed one or more new vertebral fractures since the end of the STOP-trial. The majority of fractures was wedge shaped and the deformities were intermediate to severe in both the former alendronate and alfacalcidol group. Multiple logistic regression analysis showed that STOP-trial medication and presence of pre-existing fractures did not predict development of new fractures, whereas age and cumulative glucocorticoid-dose did. During the follow-up 2.7 years after the STOP-trial both in the former alendronate and alfacalcidol group 24% of the patients underwent at least one new vertebral fracture. This underlines that prevention of vertebral fractures remains a clinical challenge, even when anti-osteoporosis drugs are prescribed.


A Dilution Method to Mitigate Biotin Interference in Cardiac Troponin T Testing.
Oral biotin supplementation is known to interfere with biotin-streptavidin-based immunoassays, including Roche's fifth-generation cardiac troponin T (cTnT) assay, which plays a critical role in the diagnosis of myocardial infarction (MI). The utility of dilution, a quick and easy method to detect and remove interferences, has not been published for biotin interference. Concentrations of cTnT were measured in pooled serum from clinical samples. Serum samples were supplemented with biotin to known concentrations, then cTnT concentrations were remeasured to assess for biotin interference. Samples were then diluted to assess for effective removal of biotin interference. At cTnT values near the critical reporting range for our institution (100 ng/L) we observed significant interference in measured values with added biotin concentrations above 50 ng/mL. In specimens without added biotin, autodilution at a 1:10 ratio yielded a mean 157% capture of measured cTnT, precluding the use of autodilution for detecting and mitigating biotin interference. A 1:10 dilution with serum containing 20-30 ng/L cTnT yielded a mean capture of 107%, which was suitable for detecting underlying biotin interference in supplemented samples. Biotin interference, at supraphysiologic concentrations, may create an artifactual reduction in measured cTnT to levels that could lead to delayed detection of an MI. Dilution with serum of known cTnT concentration of 20-30 ng/L is a fast and effective method to mitigate the analytical consequences of biotin interference.


Angioid streaks. I. Ophthalmoscopic variations and diagnostic problems.
Fifty-six patients with angioid streaks were evaluated ophthalmologically. Most had repeated fundus photography and fluorescein angiography during a follow-up period of 6 months to 7 years. The ophthalmoscopic variations and diagnostic difficulties which occurred were noted. In most instances, the angioid streaks were not initially recognized and the patient was referred with another diagnosis. In several cases, the peripapillary, macular, and peripheral changes seen with angioid streaks were found to simulate other better known fundus conditions, resulting in the erroneous diagnosis and improper treatment. In some cases, the angioid streaks were so subtle that they were overlooked and in others they were observed, but initially interpreted as something else. Because of the medical significance of angioid streaks, ophthalmologists should be aware of their variable features. These are discussed, with emphasis upon those subtleties which differentiate angioid streaks from other conditions which they may simulate. On the basis of these observations, an ophthalmoscopic differential diagnosis of angioid streaks is proposed.


Synthesis, characterization and investigation of the photophysical and photochemical properties of highly soluble novel metal-free, zinc(II), and indium(III) phthalocyanines substituted with 2,3,6-trimethylphenoxy moieties.
This work presents the synthesis and characterization of metal-free, zinc(II), and indium(III)acetate phthalocyanines substituted with 2,3,6-trimethylphenoxy groups at the peripheral and non-peripheral positions. The photophysical (fluorescence quantum yields and lifetimes) and photochemical (singlet oxygen generation and photodegradation under light irradiation) properties of these novel phthalocyanines and unsubstituted zinc(II) and indium(III)acetate phthalocyanines were investigated in dimethylformamide solution. The effects of the types of substituents and their positions and the variety of central metal ions on the phthalocyanine core on their spectroscopic, photophysical and photochemical properties were also determined. The studied 2,3,6-trimethylphenoxy substituted metal-free, zinc(II) and indium(III)acetate phthalocyanines especially indium(III)acetate derivatives exhibited appropriate photophysical and photochemical properties such as high singlet oxygen generation and these phthalocyanines can be potential Type II photosensitizers for photodynamic therapy in cancer applications.


Diagnostic value of thyrotrophin releasing hormone tests in elderly patients with atrial fibrillation.
A prospective study was carried out to compare clinical and biochemical thyroid states with responses of thyroid stimulating hormone (TSH) to thyrotrophin releasing hormone (TRH) in elderly patients with either atrial fibrillation (n = 75; mean age (SD) 79.3 (6.0) years) or sinus rhythm (n = 73; mean age 78.4 (5.6) years) admitted consecutively to the department of geriatric medicine. No patient in either group had symptoms or signs of hyperthyroidism. Overall, the TSH responses to TRH did not differ significantly between the two groups. Ten (13%) of the patients with atrial fibrillation (of whom four had raised thyroid hormone concentrations) and five (7%) of the patients with sinus rhythm showed no TSH response to TRH while 26% of each group (20 and 19 patients, respectively) showed a much reduced response. Only one of 13 patients with apparently isolated atrial fibrillation showed no TSH response to TRH, and none of these 13 patients was hyperthyroid. In particular, three patients (two with atrial fibrillation and one with sinus rhythm) who showed no TSH response to TRH at presentation exhibited a return of TSH response to TRH at follow up six weeks later. In conclusion, reduced or absent TSH responses to TRH are common in sick elderly patients whether they have atrial fibrillation or sinus rhythm and whether they are euthyroid or hyperthyroid biochemically. An absence of response is therefore an uncertain marker of hyperthyroidism in these groups of patients, and diagnosis and ablative treatment should be based at least on the presence of raised circulating free triiodothyronine or free thyroxine concentrations, or both.


Lynch Syndrome Limbo: Patient Understanding of Variants of Uncertain Significance.
Providers and patients encounter challenges related to the management of Variants of Unknown Significance (VUS). A VUS introduces new counseling dilemmas for the understanding and psychosocial impact of uncertain genetic test results. This descriptive study uses Mishel's theory of uncertainty in illness to explore the experience of individuals who have received a VUS as part of the genetic testing process. Semi-structured interviews were conducted with 27 adult individuals who received a VUS for Lynch syndrome mismatch repair genes between 2002 and 2013. The interviews were transcribed and analyzed. Most individuals recalled their result and perceived various types of uncertainty associated with their VUS. Half of the participants appraised their variant as a danger and implemented coping strategies to reduce the threat of developing cancer. Mobilizing strategies to reduce their risk included vigilant cancer surveillance, information seeking and notifying relatives. The majority of participants were unaware of the possibility of a VUS before receiving their result and expected reclassification over time. These results provide insight into the ways healthcare providers can support patients who receive VUS for Lynch syndrome. Findings also provide direction for future work that can further explicate the impact of receiving a VUS.


Vaccine schedule recommendations and updates for patients with hematologic malignancy post-hematopoietic cell transplant or CAR T-cell therapy.
Revaccination after receipt of a hematopoietic cell transplant (HCT) or cellular therapies is a pillar of patient supportive care, with the potential to reduce morbidity and mortality linked to vaccine-preventable infections. This review synthesizes national, international, and expert consensus vaccination schedules post-HCT and presents evidence regarding the efficacy of newer vaccine formulations for pneumococcus, recombinant zoster vaccine, and coronavirus disease 2019 in patients with hematological malignancy. Revaccination post-cellular therapies are less well defined. This review highlights important considerations around poor vaccine response, seroprevalence preservation after cellular therapies, and the optimal timing of revaccination. Future research should assess the immunogenicity and real-world effectiveness of new vaccine formulations and/or vaccine schedules in patients post-HCT and cellular therapy, including analysis of vaccine response that relates to the target of cellular therapies.


Impaired Cl- extrusion in layer V pyramidal neurons of chronically injured epileptogenic neocortex.
In the mature brain, the K(+)/Cl- cotransporter KCC2 is important in maintaining low [Cl-]i, resulting in hyperpolarizing GABA responses. Decreases in KCC2 after neuronal injuries result in increases in [Cl-]i and enhanced neuronal excitability due to depolarizing GABA responses. We used the gramicidin perforated-patch technique to measure E(Cl) ( approximately E(GABA)) in layer V pyramidal neurons in slices of partially isolated sensorimotor cortex of adult rats to explore the potential functional consequence of KCC2 downregulation in chronically injured cortex. E(GABA) was measured by recording currents evoked with brief GABA puffs at various membrane potentials. There was no significant difference in E(Cl) between neurons in control and undercut animals (-71.2 +/- 2.6 and -71.8 +/- 2.8 mV, respectively). However, when loaded with Cl- by applying muscimol puffs at 0.2 Hz for 60 s, neurons in the undercut cortex had a significantly shorter time constant for the positive shift in E(Cl) during the Cl- loading phase (4.3 +/- 0.5 s for control and 2.2 +/- 0.4 s for undercut, P < 0.01). The positive shift in E(Cl) 3 s after the beginning of Cl- loading was also significantly larger in the undercut group than in the control, indicating that neurons in undercut cortex were less effective in maintaining low [Cl-]i during repetitive activation of GABA(A) receptors. Application of furosemide eliminated the difference between the control and undercut groups for both of these measures of [Cl-]i regulation. The results suggest an impairment in Cl- extrusion resulting from decreased KCC2 expression that may reduce the strength of GABAergic inhibition and contribute to epileptogenesis.


Design and Synthesis of Novel Cereblon Binders for Use in Targeted Protein Degradation.
Modulating the chemical composition of cereblon (CRBN) binders is a critical step in the optimization process of protein degraders that seek to hijack the function of this E3 ligase. Small structural changes can have profound impacts on the overall profile of these compounds, including depth of on-target degradation, neosubstrate degradation selectivity, as well as other drug-like properties. Herein, we report the design and synthesis of a series of novel CRBN binding moieties. These CRBN binders were evaluated for CRBN binding and degradation of common neosubstrates Aiolos and GSPT1. A selection of these binders was employed for an exploratory matrix of heterobifunctional molecules, targeting CRBN-mediated degradation of the androgen receptor.


Novel imatinib derivatives with altered specificity between Bcr-Abl and FMS, KIT, and PDGF receptors.
Imatinib is a clinically important ATP analogue inhibitor that targets the tyrosine kinase domain of the intracellular Abl kinase and the PDGF receptor family. Imatinib has revolutionised the treatment of chronic myeloid leukaemia, which is caused by the oncogene Bcr-Abl and certain solid tumours that harbor oncogenic mutations of the PDGF receptor family. As a leading kinase inhibitor, imatinib also provides an excellent model system to investigate how changes in drug design impact biological activity, which is an important consideration for rational drug design. Herein we report a new series of imatinib derivatives that in general have greater activity against the family of PDGF receptors and poorer activity against Abl, as a result of modifications of the phenyl and N-methylpiperazine rings. These new compounds provide a platform for further drug development against the therapeutically important PDGF receptor family and they also provide insight into the engineering of drugs with altered biological activity.


Developmentally regulated MAPK pathways modulate heterochromatin in Saccharomyces cerevisiae.
Variegated expression of genes contributes to phenotypic variation within populations of genetically identical cells. Such variation plays a role in development and host pathogen interaction and can be important in adaptation to harsh environments. The expression state of genes placed near telomeres shows a variegated pattern of inheritance due to heterochromatin formation, a phenomenon that is called telomere position effect (TPE). We show that in budding yeast, TPE is controlled by the a1/alpha2 developmental repressor, which dictates developmental decisions in response to environmental changes. Two a1/alpha2 repressed genes, STE5, a MAPK scaffold and HOG1, a stress-activated MAPK, are the targets of this heterochromatin regulation pathway. We provide new evidence that link MAPK signaling and heterochromatin formation in yeast. Our results show that the same components that regulate gene expression states in euchromatic regions regulate heterochromatic expression states and that stress can play a part in turning on or off genes placed in heterochromatic regions.