3D mouse embryonic stem cell culture for generating inner ear organoids.
This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 96-well plates in medium containing extracellular matrix proteins to promote epithelialization. During the first 14 d, a series of precisely timed protein and small-molecule treatments sequentially induce epithelia that represent the mouse embryonic non-neural ectoderm, preplacodal ectoderm and otic vesicle epithelia. Ultimately, these tissues develop into cysts with a pseudostratified epithelium containing inner ear hair cells and supporting cells after 16-20 d. Concurrently, sensory-like neurons generate synapse-like structures with the derived hair cells. We have designated the stem cell-derived epithelia harboring hair cells, supporting cells and sensory-like neurons as inner ear organoids. This method provides a reproducible and scalable means to generate inner ear sensory tissue in vitro.


Emphysema and its relation to age.
To establish whether a correlation existed between emphysema and age, 24 lungs were used to measure the alveolar membrane fenestrations; the average area of 100 fenestrae was then plotted against age. Eighteen of these lungs were from patients 17 to 86 years of age with no history of respiratory ailments, whereas 6 lungs were from patients in whom an antemortem diagnosis of emphysema had been made. A significant, positive correlation between age and the area of fenestrae (t=2.4;p greater than 0.05) was found in the group of 18 lungs from patients with no respiratory ailements. The same could not be said categorically about the lungs from the emphysematous group, because the number of lungs examined was too small. We postulate that the pores of Kohn are part of the degenerating emphysematous process and are the first stage in the formation of fenestrae. A possible mechanism for the formation of fenestrae and their classification into 4 grades is presented.


Chromosomal fragility in the cancer-prone disease xeroderma pigmentosum preferentially involves bands relevant for cutaneous carcinogenesis.
Spontaneous and folate-induced chromosomal fragility was analyzed in peripheral blood lymphocytes from 6 patients affected by the cancer-prone disease xeroderma pigmentosum (XP), from the parents of 4 of the patients, and from 12 normal subjects. All XP patients were defective in nucleotide-excision repair; 4 belonged to group C and 1 each to groups A and D. A tendency toward increased spontaneous chromosomal fragility was observed in the XP family members, and lesions indicating substantial chromosomal damage, which were not observed in any healthy donors, were frequently found. The spontaneous lesion sites in lymphocytes from homozygous and heterozygous carriers of XP defects appeared to be significantly associated with those observed in normal skin fibroblasts from the same subjects. These XP spontaneous fragility sites showed a statistically significant association with the rearrangement breakpoints reported in skin pre-tumoral and tumoral lesions from normal and unrelated XP donors. Under conditions of folate deprivation, the chromosomal fragility level, the pattern and the frequency of expression of fragile sites in XP patients and in their parents were similar to normal. However, XP patients generally showed a higher susceptibility to breakage at sites described as mutagen and carcinogen targets.


Disruption of MAP-2 immunostaining in rat hippocampus after traumatic brain injury.
The effects of diffuse brain injury on dendritic morphology in rat hippocampus and cortex were examined in this study using the recently described impact acceleration model of traumatic brain injury (Marmarou et al., 1994). Dendritic structure was visualized using immunostaining of microtubule associated protein-2 (MAP-2). Brains were studied 24, 48, and 72 h after brain injury. Results from immunohistochemistry and light microscopy indicated a time-dependent disruption of dendritic cytoarchitecture in the CA1 subregion and in the hilus of the hippocampus but not in the dentate gyrus or CA3 subregion. Similar disruption was observed in the cortical mantle overlying the hippocampus. Although disruption of dendritic structure was observed at 24 h, the most severe damage was at 48 h after injury with evidence of at least partial recovery of MAP-2 immunostaining by 72 h. In the most severe damage, dendrites appeared to be fragmented, scattered, and unaligned, consisting of irregularly spaced and darkly stained swollen segments. A mixed pattern of immunostaining was observed in somata of hilar cells, with some appearing normal while others stained only faintly, appearing to have lost their typical polygonal shape. Semiquantitative rankings confirmed these qualitative findings. Immediate post-injury behavioral evaluations of injury severity were compared to the degree of disruption of MAP-2 immunostaining. The results of this study indicate that diffuse brain injury is associated not only with axonal damage but also with injury to dendrites.


17 beta-hydroxysteroid dehydrogenase type XI localizes to human steroidogenic cells.
We searched expressed sequence tag databases with conserved domains of the short-chain alcohol dehydrogenase superfamily and identified another isoform of 17 beta-hydroxysteroid dehydrogenase, 17 beta HSDXI. This enzyme converts 5 alpha-androstane-3 alpha, 17 beta-diol to androsterone. The substrate has been implicated in supporting gestation and modulating gamma-aminobutyric acid receptor activity. 17 beta HSDXI is colinear with human retinal short-chain dehydrogenase/reductase retSDR2, a protein with no known biological activity (accession no. AAF06939). Of the proteins with known function, 17 beta HSDXI is most closely related to the retinol-metabolizing enzyme retSDR1, with which it has 30% identity. There is a polymorphic stretch of 15 adenosines in the 5' untranslated region of the cDNA sequence and a silent polymorphism at C719T. A 17 beta HSDXI construct with a stretch of 20 adenosines was found to produce significantly more enzyme activity than constructs containing 15 or less adenosines (43% vs. 26%, P < 0.005). The C719T polymorphism is present in 15% of genomic DNA samples. Northern blot analysis showed high levels of 17 beta HSDXI expression in the pancreas, kidney, liver, lung, adrenal, ovary, and heart. Immunohistochemical staining for 17 beta HSDXI is strong in steroidogenic cells such as syncytiotrophoblasts, sebaceous gland, Leydig cells, and granulosa cells of the dominant follicle and corpus luteum. In the adrenal 17 beta HSDXI, staining colocalized with the distribution of 17 alpha-hydroxylase but was stronger in the mid to outer cortex. 17 beta HSDXI was also found in the fetus and increased after birth. Liver parenchymal cells and epithelium of the endometrium and small intestine also stained. Regulation studies in mouse Y1 cells showed that cAMP down-regulates 17 beta HSDXI enzymatic activity (40% vs. 32%, P < 0.05) and reduces gene expression to undetectable levels. All-trans-retinoic acid did not affect 17 beta HSDXI expression or activity, but addition of the retinoid together with cAMP significantly decreased activity over cAMP alone (32% vs. 23%, P < 0.05). Cloning and sequencing of the 17 beta HSDXI promoter identified the potential nuclear receptor steroidogenic factor-1 half-site TCCAAGGCCGG, and a cluster of three other potential steroidogenic factor-1 half-sites were found in the distal part of intron 1. Collectively, these results suggest a role for 17 beta HSDXI in androgen metabolism during steroidogenesis and a possible role in nonsteroidogenic tissues including paracrine modulation of 5 alpha-androstane-3 alpha, 17 beta-diol levels. 17 beta HSDXI could act by metabolizing compounds that stimulate steroid synthesis and/or by generating metabolites that inhibit it.


European consensus statement on the terminology used in the management of lupus glomerulonephritis.
Systemic lupus erythematosus (SLE) is a complex, multisystem autoimmune disorder, which often involves referral to multiple medical specialists. Lupus nephritis (LN) occurs in ~35% of adults with SLE and predicts poor survival. There is currently no consensus on how to manage patients with SLE or LN across specialties and across different European countries. The Lupus Nephritis Terminology Advisory Group was formed to address this issue as it impacts upon LN treatment. It has developed consensus statements based on opinions from expert panel meetings with nephrologists, nephropathologists, rheumatologists, clinical immunologists and internal medicine specialists from many European countries, after reviewing current guidelines from the European League Against Rheumatism, the American College of Rheumatology and the participants' experience. In this article, we report consensus statements that were developed in six important areas: classification of patients with LN, how classification affects the selection of treatment options and definitions of induction, response, flare and maintenance. We have also proposed a consensus for the terminology involved in the management of LN that is consistent with clinical opinion gathered from multidisciplinary expert meetings and with existing guidelines. We believe this consensus approach provides agreed expert opinion to clinicians and will form the basis for optimising LN treatment.


[The investigation of the contacts of the tuberculous pediatric patient].
Results of contacts investigation of tuberculous pediatric patients, usually non-infectious, are shown. Their relationship to the clinical situation on index case and the intimacy of exposure is analyzed. 714 contacts of 111 pediatric cases (53 category II and 58 category III, according to the American Thoracic Society classification). The tuberculin skin test with 2 units of PPD-RT23 was used to identify the infected persons. Chest radiography, sputum smear and culture examination were used to diagnose current pulmonary disease. According to the intimacy of exposure to index case, contacts were defined as household, close or sporadic contacts. The possible association with clinical situation of index case and intimacy of exposure was analyzed. 41 new cases of tuberculosis were detected, 7 of them were infectious. Prevalence of tuberculous infection among contacts was 44%. Quimioprofilaxis was indicated to 126 contacts. The prevalences of infection and disease according to clinical situation and degree of contact did not differ significantly. it is advisable to investigate the contacts of tuberculous pediatric patients, even in cases of tuberculous infection without disease; the investigation must include close and sporadic contacts.


A critical review of experimental and predicted methane generation from anaerobic codigestion.
Anaerobic digestion is increasingly being considered as a treatment option for an extensive range of waste biomass, due to the potential for energy recovery, in the form of methane production, and lower sludge volumes relative to aerobic treatment processes. Furthermore, when two substrates are codigested (i.e. digested together), added benefits are foreseeable, such as increased methane production and detoxification of toxic compounds via cometabolic degradation pathways. The objectives of this study were to compare experimental and predicted methane production from codigestion literature studies in order to objectively evaluate digester performance. Two predictive methods were used, both assuming methane yields are additive: literature values for digestion of single substrates and a stoichiometric method using model substrates to represent different substrates. Waste sources included in the analysis were primary sewage sludge, waste activated sludge, cow manure, waste paper, grease trap sludge, fat oil and grease and algal sludge. It was found that methane production could approximately be predicted using both methods, with literature methane yields from the same study being the most accurate predictor. One important finding from this study was that the assumption that methane yields are additive is a reasonable one. Furthermore, both predictive methods may be usefully employed as a screening tool to compare methane yields between different types and blends of substrates.


[Growth hormone usage in Prader-Willi syndrome].
Prader-Willi syndrome (PWS), with a prevalence of 60:1.000.000, results from the loss of paternal chromosome 15, being 56% due to deletion, 24% due to uniparental maternal disomy, and 18% from methylation, an epigenetic phenomenon. The clinical picture begins with extreme muscular hypotonia, which makes it difficult to feed the child in the first year. As the hypotonia improves, usually in the first two years, around the 4th year of life, an insatiable appetite leads these children to an extreme obesity, with alveolar hypoventilation which endangers their lives. So, paradoxically, PWS threatens the lives of the patients, through inanition in a first phase and, afterwards, through excessive weight gain. The use of growth hormone (hrGH) in these children has a primary goal to change the body composition and improve the physical activity and the quality of life. On the other hand, many PWS patients are indeed GH deficient, and an improvement in the height SDS occurs with treatment. We have to be careful, however. When starting a PWS treatment with a patient on hrGH, a careful evaluation of sleep apnoea (polysomnography) as well as a careful examination of the airways is extremely mandatory, since the treatment may compromise the respiratory pattern of some patients.


Eosinophilia and Kidney Disease: More than Just an Incidental Finding?
Peripheral blood eosinophilia (PBE), defined as 500 eosinophils or above per microliter (µL) blood, is a condition that is not uncommon but often neglected in the management of patients with chronic kidney disease (CKD), acute kidney injury (AKI), or patients on renal replacement therapy (RRT). The nature of PBE in the context of kidney diseases is predominantly secondary or reactive and has to be distinguished from primary eosinophilic disorders. Nonetheless, the finding of persistent PBE can be a useful clue for the differential diagnosis of underdiagnosed entities and overlapping syndromes, such as eosinophilic granulomatosis with polyangiitis (EGPA), IgG4-related disease (IgG4-RD), acute interstitial nephritis (AIN), or the hypereosinophilic syndrome (HES). For patients on RRT, PBE may be an indicator for bio-incompatibility of the dialysis material, acute allograft rejection, or Strongyloides hyperinfection. In a subset of patients with EGPA, eosinophils might even be the driving force in disease pathogenesis. This improved understanding is already being used to facilitate novel therapeutic options. Mepolizumab has been licensed for the management of EGPA and is applied with the aim to abrogate the underlying immunologic process by blocking interleukin-5. The current article provides an overview of different renal pathologies that are associated with PBE. Further scientific effort is required to understand the exact role and function of eosinophils in these disorders which may pave the way to improved interdisciplinary management of such patients.


Transgenic-cloned pigs systemically expressing red fluorescent protein, Kusabira-Orange.
Genetically engineered pigs with cell markers such as fluorescent proteins are highly useful in lines of research that include the tracking of transplanted cells or tissues. In this study, we produced transgenic-cloned pigs carrying a gene for the newly developed red fluorescent protein, humanized Kusabira-Orange (huKO), which was cloned from the coral stone Fungia concinna. The nuclear transfer embryos, reconstructed with fetal fibroblast cells that had been transduced with huKO cDNA using retroviral vector D Delta Nsap, developed efficiently in vitro into blastocysts (28.0%, 37/132). Nearly all (94.6%, 35/37) of the cloned blastocysts derived from the transduced cells exhibited clear huKO gene expression. A total of 429 nuclear transfer embryos were transferred to four recipients, all of which became pregnant and gave birth to 18 transgenic-cloned offspring in total. All of the pigs highly expressed huKO fluorescence in all of the 23 organs and tissues analyzed, including the brain, eyes, intestinal and reproductive organs, skeletal muscle, bone, skin, and hoof. Furthermore, such expression was also confirmed by histological analyses of various tissues such as pancreatic islets, renal corpuscles, neuronal and glial cells, the retina, chondrocytes, and hematopoietic cells. These data demonstrate that transgenic-cloned pigs exhibiting systemic red fluorescence expression can be efficiently produced by nuclear transfer of somatic cells retrovirally transduced with huKO gene.


Fracture rate after conventional external beam radiation therapy to the spine in multiple myeloma patients.
Conventional external beam radiation therapy (cEBRT) is used in multiple myeloma (MM) to treat severe pain, spinal cord compression, and disease-related bone disease. However, radiation may be associated with an increased risk of vertebral compression fractures (VCFs), which could substantially impair survival and quality of life. Additionally, the use of the Spinal Instability Neoplastic Score (SINS) in MM is debated in MM. To determine the incidence of VCFs after cEBRT in patients with MM and to assess the applicability of the SINS score in the prediction of VCFs in MM. Retrospective multicenter cohort study. MM patients with spinal myeloma lesions who underwent cEBRT between January 2010 and December 2021. Frequency of new or progressed VCFs and subdistribution hazard ratios for potentially associated factors. Patient and treatment characteristics were manually collected from the patients' electronic medical records. Computed tomography (CT) scans from before and up to 3 years after the start of radiation were used to score radiographic variables at baseline and at follow-up. Multivariable Fine and Gray competing risk analyses were performed to evaluate the diagnostic value of the SINS score to predict the post-radiation VCF rate. A total of 127 patients with 427 eligible radiated vertebrae were included in this study. The mean age at radiation was 64 years, and 66.1% of them were male. At the start of radiation, 57 patients (44.9%) had at least one VCF. There were 89 pre-existing VCFs (18.4% of 483 vertebrae). Overall, 39 of 127 patients (30.7%) reported new fractures (number of vertebrae (n)=12) or showed progression of existing fractures (n=36). This number represented 11.2% of all radiated vertebrae. Five of the 39 (12.8%) patients with new or worsened VCFs received an unplanned secondary treatment (augmentation [n=2] or open surgery [n=3]) within 3 years. Both the total SINS score (SHR 1.77; 95% confidence interval (CI) 1.54-2.03; p<.001) and categorical SINS score (SHR 10.83; 95% CI 4.20-27.94; p<.001) showed an independent association with higher rates of new or progressed VCFs in adjusted analyses. The use of bisphosphonates was independently associated with a lower rate of new or progressed VCFs (SHR 0.47 [95% CI 0.24-0.92; p=.027]). This study demonstrated that new or progressed VCFs occurred in 30.7% of patients within 3 years, in a total of 11.2% of vertebrae. The SINS score was found to be independently associated with the development or progression of VCFs and could thus be applied in MM for fracture prediction and possibly prevention.


The facilitated early enteral and dietary management effectiveness trial in hospitalized patients with malnutrition.
The prevalence of malnutrition in the hospitalized setting is 30% to 55%. Previous studies reported an association of malnutrition with an increased hospital length of stay (LOS), morbidity, and mortality of patients. This study evaluated the role of early nutrition intervention on LOS, diagnosis coding of malnutrition cases, calculating case mix index, and reducing delays in implementing nutrition support to patients. Demographic data, anthropometric measurements, LOS, and serum albumin levels were collected from 400 patients in 2 medical wards to determine the prevalence of malnutrition and potential delays in nutrition consultation. Based on these results, a nutrition intervention study was conducted in 1 ward; the other ward served as a control. Patients were classified as normally nourished or malnourished. Multivariate general linear regressions were used to reveal the impact of intervention on the change in LOS, controlling for other potential confounding factors on the cohort and a subset with severe malnutrition. Of the 400 patients assessed, 53% had malnutrition. Multiple general linear regressions showed that nutrition intervention reduced LOS an average of 1.93 days in the cohort group and 3.2 days in the severe malnourished group. Case mix index and female gender were positively associated with LOS in the malnourished group. Nutrition intervention reduced the delays in implementing nutrition support to patients by 47%. Results highlight the positive impact of nutrition intervention in terms of reduced LOS in malnourished hospital patients. Reduction in LOS with diagnosis coding of malnutrition cases yielded substantial economic benefits.


Carbon monoxide-activated Nrf2 pathway leads to protection against permanent focal cerebral ischemia.
Carbon monoxide (CO) is a gaseous second messenger produced when heme oxygenase enzymes catabolize heme. We have demonstrated that CO can be therapeutic in ischemia-reperfusion brain injury; however, it is unclear whether CO can also offer protection in permanent ischemic stroke or what mechanism(s) underlies the effect. Heme oxygenase-1 neuroprotection was shown to be regulated by Nrf2; therefore, we investigated whether CO might partially exert neuroprotection by modulating the Nrf2 pathway. To evaluate the potential protective effects of CO, we exposed male wild-type and Nrf2-knockout mice to 250 ppm CO or control air for 18 hours immediately after permanent middle cerebral artery occlusion. Infarct volume and neurologic deficits were assessed on day 7. Molecular mechanisms of Nrf2 pathway activation by CO were also investigated. Mice exposed to CO after permanent ischemia had 29.6±12.6% less brain damage than did controls at 7 days, although amelioration in neurologic deficits did not reach significance. Additionally, 18-hour CO treatment led to Nrf2 dissociation from Keap1, nuclear translocation, increased binding activity of Nrf2 to heme oxygenase-1 antioxidant response elements, and elevated heme oxygenase-1 expression 6 to 48 hours after CO exposure. The CO neuroprotection was completely abolished in Nrf2-knockout mice. Low-concentration CO represent a neuroprotective agent for combination treatment of ischemic stroke, and its beneficial effect would be at least partially mediated by activation of the Nrf2 pathway.


The epitope specificity of heparin-induced thrombocytopenia.
Heparin-induced thrombocytopenia (HIT) is caused by antibodies (HIT-Abs) that bind to a complex of heparin and platelet factor 4. We investigated the epitope specificity of the HIT-Abs, and found that the HIT-Abs recognized solid-phase immobilized complexes with an optimum ratio of four to eight molecules of PF4 per molecule of heparin. To try to define the epitopes within the PF4 molecule, intact and reduced (linearized) PF4 was tested against 29 different sera from patients with HIT. In addition, eight different peptides that spanned the PF4 molecule were studied for their ability to bind to the HIT-Abs either alone or in the presence of heparin. With the exception of a subpopulation of patient samples (5/29, 17%), we found that reduced PF4 and the peptides were uniformly non-reactive with the HIT-Abs in the presence of heparin. Reduced PF4 and PF4 carboxy-terminal peptides with a minimum size of 19 amino acids were recognized by a minority (5/29) of HIT-Abs samples but only when heparin was present. The specificity of this subgroup of samples from patients with HIT was highly restricted and the loss of one amino acid (peptide reduced in length from 19 to 18 amino acids) rendered the peptides non-reactive. The clinical characteristics of these patients were similar to the other HIT patients. These studies demonstrate that the majority of HIT-Abs recognize a noncontiguous conformational epitope on the PF4 molecule that is produced when four to eight PF4 molecules are bound together by heparin.


Human SCARB2-dependent infection by coxsackievirus A7, A14, and A16 and enterovirus 71.
Human enterovirus species A (HEV-A) consists of at least 16 members of different serotypes that are known to be the causative agents of hand, foot, and mouth disease (HFMD), herpangina, and other diseases, such as respiratory disease and polio-like flaccid paralysis. Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the major causative agents of HFMD. CVA5, CVA6, CVA10, and CVA12 mainly cause herpangina or are occasionally involved with sporadic cases of HFMD. We have previously shown that human scavenger receptor class B, member 2 (SCARB2) is a cellular receptor for EV71 and CVA16. Using a large number of clinical isolates of HEV-A, we explored whether all clinical isolates of EV71 and other serotypes of HEV-A infected cells via SCARB2. We tested this possibility by infecting L-SCARB2 cells, which are L929 cells expressing human SCARB2, by infecting human RD cells that had been treated with small interfering RNAs for SCARB2 and by directly binding the viruses to a soluble SCARB2 protein. We showed that all 162 clinical isolates of EV71 propagated in L-SCARB2 cells, suggesting that SCARB2 is the critical receptor common to all EV71 strains. In addition, CVA7, CVA14, and CVA16, which are most closely related to each other, also utilized SCARB2 for infection. EV71, CVA14, and CVA16 are highly associated with HFMD, and EV71 and CVA7 are occasionally associated with neurological diseases, suggesting that SCARB2 plays important roles in the development of these diseases. In contrast, another group of viruses, such as CVA2, CVA3, CVA4, CVA5, CVA6, CVA8, CVA10, and CVA12, which are relatively distant from the EV71 group, is associated mainly with herpangina. None of these clinical isolates infected via the SCARB2-dependent pathway. HEV-A viruses can be divided into at least two groups depending on the use of SCARB2, and the receptor usage plays an important role in developing the specific diseases for each group.


Variations in clinical nurse leaders' confidence with performing the core role functions.
Clinical nurse leader (CNL) practice, by definition, requires individuals to make career transitions. CNLs must adjust to their new work role and responsibilities and doing so also entails individual adjustment. Prior work has not examined the role of individual-level factors in effective CNL role transition. This study contributes to CNL implementation efforts by developing understanding of personal and contextual factors that explain variation in individuals' levels of self-confidence with performing the key functions of the CNL role. Data were gathered using a cross-sectional survey from a national sample of registered nurses (RNs) certified as CNLs. Respondents' perceptions of their confidence in performing CNL role competencies were measured with the Clinical Nurse Leader Self-Efficacy Scale (CNLSES; Gilmartin MJ, Nokes, K. (in press). The Clinical Nurse Leader Self Efficacy Scale: Results of a pilot study. Nursing Economic$). The CNLSES is a 35-item state-specific self-efficacy scale with established measurement properties that assesses nurses' perceptions of their ability to function effectively as a CNL. Demographic data were also collected. Data were analyzed using a general linear regression model. One hundred forty-seven certified CNLs participated in the survey. Results indicate that respondents vary in their confidence with performing the nine role competencies associated with CNL practice. Results from regression analyses also show that respondents' confidence in their abilities to carry out the core functions associated with the CNL role varied significantly across geographic region, organizational type, and by CNL graduate program model. The results of this study show important differences in CNLs' levels of self-confidence with the core competencies of their role. As a result, it may be important to develop targeted career transition interventions to gain the full benefit of CNL practice.


Expression of Hormonal Carcinogenesis Genes and Related Regulatory microRNAs in Uterus and Ovaries of DDT-Treated Female Rats.
The insecticide dichlorodiphenyltrichloroethane (DDT) is a nonmutagenic xenobiotic compound able to exert estrogen-like effects resulting in activation of estrogen receptor-α (ERα) followed by changed expression of its downstream target genes. In addition, studies performed over recent years suggest that DDT may also influence expression of microRNAs. However, an impact of DDT on expression of ER, microRNAs, and related target genes has not been fully elucidated. Here, using real-time PCR, we assessed changes in expression of key genes involved in hormonal carcinogenesis as well as potentially related regulatory oncogenic/tumor suppressor microRNAs and their target genes in the uterus and ovaries of female Wistar rats during single and chronic multiple-dose DDT exposure. We found that applying DDT results in altered expression of microRNAs-221, -222, -205, -126a, and -429, their target genes (Pten, Dicer1), as well as genes involved in hormonal carcinogenesis (Esr1, Pgr, Ccnd1, Cyp19a1). Notably, Cyp19a1 expression seems to be also regulated by microRNAs-221, -222, and -205. The data suggest that epigenetic effects induced by DDT as a potential carcinogen may be based on at least two mechanisms: (i) activation of ERα followed by altered expression of the target genes encoding receptor Pgr and Ccnd1 as well as impaired expression of Cyp19a1, affecting, thereby, cell hormone balance; and (ii) changed expression of microRNAs resulting in impaired expression of related target genes including reduced level of Cyp19a1 mRNA.


Online mass spectrometry of CE (SDS)-separated proteins by two-dimensional capillary electrophoresis.
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is the fundamental technique for protein separation by size. Applying this technology in capillary format, gaining high separation efficiency in a more automated way, is a key technology for size separation of proteins in the biopharmaceutical industry. However, unequivocal identification by online mass spectrometry (MS) is impossible so far, due to strong interference in the electrospray process by SDS and other components of the SDS-MW separation gel buffer. Here, a heart-cut two-dimensional electrophoretic separation system applying an electrically isolated valve with an internal loop of 20 nL is presented. The peak of interest in the CE (SDS) separation is transferred to the CZE-MS, where electrospray-interfering substances of the SDS-MW gel are separated prior to online electrospray ionization mass spectrometry. An online SDS removal strategy for decomplexing the protein-SDS complex is implemented in the second dimension, consisting of the co-injection of organic solvent and cationic surfactant. This online CE (SDS)-CZE-MS system allows MS characterization of proteoforms separated in generic CE (SDS), gaining additional separation in the CZE and detailed MS information. In general, the system can be applied to all kinds of proteins separated by CE (SDS). Here, we present results of the CE (SDS)-CZE-MS system on the analysis of several biopharmaceutically relevant antibody impurities and fragments. Additionally, the versatile application spectrum of the system is demonstrated by the analysis of extracted proteins from soybean flour. The online hyphenation of CE (SDS) resolving power and MS identification capabilities will be a powerful tool for protein and mAb characterization. Graphical abstract Two-dimensional capillary electrophoresis system hyphenated with mass spectrometry for the characterization of CE (SDS)-separated proteins. As first dimension, a generic and high MS-interfering CE (SDS) separation is performed for size separation. After heart-cut transfer of the unknown CE (SDS) protein peak, via a four-port nanoliter valve to a volatile electrolyte system as second dimension, interference-free mass spectrometric data of separated mAb fragments and soybean proteins are obtained.


Automatic Assessment of Full Left Ventricular Coverage in Cardiac Cine Magnetic Resonance Imaging with Fisher Discriminative 3D CNN.
Cardiac magnetic resonance (CMR) images play a growing role in the diagnostic imaging of cardiovascular diseases. Full coverage of the left ventricle (LV), from base to apex, is a basic criterion for CMR image quality and necessary for accurate measurement of cardiac volume and functional assessment. Incomplete coverage of the LV is identified through visual inspection, which is time-consuming and usually done retrospectively in the assessment of large imaging cohorts. This paper proposes a novel automatic method for determining LV coverage from CMR images by using Fisher-discriminative three-dimensional (FD3D) convolutional neural networks (CNNs). In contrast to our previous method employing 2D CNNs, this approach utilizes spatial contextual information in CMR volumes, extracts more representative high-level features and enhances the discriminative capacity of the baseline 2D CNN learning framework, thus achieving superior detection accuracy. A two-stage framework is proposed to identify missing basal and apical slices in measurements of CMR volume. First, the FD3D CNN extracts high-level features from the CMR stacks. These image representations are then used to detect the missing basal and apical slices. Compared to the traditional 3D CNN strategy, the proposed FD3D CNN minimizes within-class scatter and maximizes between-class scatter. We performed extensive experiments to validate the proposed method on more than 5,000 independent volumetric CMR scans from the UK Biobank study, achieving low error rates for missing basal/apical slice detection (4.9%/4.6%). The proposed method can also be adopted for assessing LV coverage for other types of CMR image data.


Over-expression of Id-1 induces cell proliferation in hepatocellular carcinoma through inactivation of p16INK4a/RB pathway.
Inhibitors of differentiation and DNA binding-1 (Id-1) have been demonstrated to oppose Ets-mediated activation of p16INK4a. As p16INK4a protein is inactivated in hepatocellular carcinoma (HCC), we aimed to investigate the role of Id-1 in regulating p16INK4a expression during the development of HCC in HCC patients and direct ectopic Id-1 introduction into the PLC/PRF/5 HCC cell line. Sixty-two HCC samples were recruited for evaluation of Id-1 and proliferating cell nuclear antigen (PCNA) protein expression. The messenger RNA (mRNA) expression of Id-1 and p16INK4a was detected by quantitative reverse transcription-polymerase chain reaction. For in vitro Id-1 transfection, five Id-1 transfected clones were isolated and the effect of ectopic Id-1 introduction was investigated by 3-(4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, flow cytometry, immunostaining and western blot. Our results showed that Id-1 was over-expressed in HCC specimens both at mRNA and protein levels. Over-expression of Id-1 protein was correlated with PCNA (r = 0.334, P = 0.033). HCC samples showing low Id-1 protein expression had a lower Id-1 mRNA level (340.2 versus 1467%, P = 0.039) and higher p16INK4a expression (195 versus -78.6%, P = 0.039) than samples with high Id-1 protein expression. In the PLC/PRF/5 HCC cell line study, ectopic Id-1 expression resulted in proliferation of HCC cells and an increased percentage of S phase cells and PCNA expression. The results showed that over-expression of Id-1 induces cell proliferation in HCC through inactivation of p16INK4a/retinoblastoma pathway. In conclusion, the results provided an insight for the understanding of the role of Id-1 in functional inactivation of p16INK4a in HCC.


Inhibition of vasoconstriction by tetramethylpyrazine: does it act by blocking the voltage-dependent Ca channel?
Using dog mesenteric arterial ring preparations, we studied the in vitro vascular effects of tetramethylpyrazine (TMP, also known as Ligustrazine), a vasoactive component derived from Ligustium Wollichii Franchat; a widely used ingredient in Chinese herbal medicine for the treatment of cardiovascular diseases. TMP had no effect on the resting force of the vascular muscle, but it caused a dose-dependent inhibition of vascular contractile responses to KCl and phenylephrine (PHE). These inhibitory effects of TMP were reversible and more prominent at lower agonist concentrations. TMP also inhibited the responses to PHE in Ca-free medium containing 50 microM EGTA suggesting the action on the release of intracellular Ca2+. Therefore, TMP is not a specific Ca-channel blocker as claimed previously. Its vasodilatory effect is mediated by the inhibition of Ca influx, as well as the release of intracellular Ca2+.


Effects of β-conglycinin on growth performance, immunoglobulins and intestinal mucosal morphology in piglets.
One of the main causes of allergic reactions in young animals is β-conglycinin, an antigenic glycoprotein found in soya beans. Therefore, the objective of the study was to investigate the effects of a prior immunisation with β-conglycinin on growth performance, serum immunoglobulin levels and intestinal histology in piglets. Forty piglets (7 d of age) were randomly divided into four groups of ten piglets each. Piglets of Groups Im and Im+S were immunised twice by hypodermic injection with β-conglycinin at 500 μg/kg body weight (BW) at day 7 and 21 of age. At day 23, Groups Im+S and S were intramuscularly injected with 5000 μg β-conglycinin per kg BW. The piglets of Group C received a physiological saline solution by hypodermic injection. All piglets were weaned at the age of 23 d and blood samples were taken on days 7, 21 and 35. At the end of the trial, five piglets per group were slaughtered and the intestine was collected for evaluating mucosal histology. Compared to Group C, in Group S the average daily gain (ADG), feed intake and gain:feed ratio were decreased (p < 0.01), and serum levels of IgG and IgE were increased (p < 0.01). Furthermore, in this group the structure of duodenal and jejunal mucosa was severely damaged. But in Groups Im and Im+S the ADG was increased (p < 0.05), serum IgE levels were decreased (p < 0.01) and the intestinal mucosa was not damaged. The results suggest that prior immunisation with β-conglycinin can increase ADG and serum IgG levels and decrease serum IgE levels. Therefore, this method is also potentially able to protect the structural integrity of the intestinal mucosal epithelia and alleviate allergic reactions in piglets.


Exploring the Nonconserved Sequence Space of Synthetic Expression Modules in Bacillus subtilis.
Increasing protein expression levels is a key step in the commercial production of enzymes. Predicting promoter activity and translation initiation efficiency based solely on consensus sequences have so far met with mixed results. Here, we addressed this challenge using a "brute-force" approach by designing and synthesizing a large combinatorial library comprising ∼12 000 unique synthetic expression modules (SEMs) for Bacillus subtilis. Using GFP fluorescence as a reporter of gene expression, we obtained a dynamic expression range that spanned 5 orders of magnitude, as well as a maximal 13-fold increase in expression compared with that of the already strong veg expression module. Analyses of the synthetic modules indicated that sequences at the 5'-end of the mRNA were the most important contributing factor to the differences in expression levels, presumably by preventing formation of strong secondary mRNA structures that affect translation initiation. When the gfp coding region was replaced by the coding region of the xynA gene, encoding the industrially relevant B. subtilis xylanase enzyme, only a 3-fold improvement in xylanase production was observed. Moreover, the correlation between GFP and xylanase expression levels was weak. This suggests that the differences in expression levels between the gfp and xynA constructs were due to differences in 5'-end mRNA folding and consequential differences in the rates of translation initiation. Our data show that the use of large libraries of SEMs, in combination with high-throughput technologies, is a powerful approach to improve the production of a specific protein, but that the outcome cannot necessarily be extrapolated to other proteins.


Predicting Deterioration from Wearable Sensor Data in People with Mild COVID-19.
Coronavirus has caused many casualties and is still spreading. Some people experience rapid deterioration that is mild at first. The aim of this study is to develop a deterioration prediction model for mild COVID-19 patients during the isolation period. We collected vital signs from wearable devices and clinical questionnaires. The derivation cohort consisted of people diagnosed with COVID-19 between September and December 2021, and the external validation cohort collected between March and June 2022. To develop the model, a total of 50 participants wore the device for an average of 77 h. To evaluate the model, a total of 181 infected participants wore the device for an average of 65 h. We designed machine learning-based models that predict deterioration in patients with mild COVID-19. The prediction model, 10 min in advance, showed an area under the receiver characteristic curve (AUC) of 0.99, and the prediction model, 8 h in advance, showed an AUC of 0.84. We found that certain variables that are important to model vary depending on the point in time to predict. Efficient deterioration monitoring in many patients is possible by utilizing data collected from wearable sensors and symptom self-reports.


The primacy of temporal dynamics in driving spatial self-organization of soil iron redox patterns.
This study investigates mechanisms that generate regularly spaced iron-rich bands in upland soils. These striking features appear in soils worldwide, but beyond a generalized association with changing redox, their genesis is yet to be explained. Upland soils exhibit significant redox fluctuations driven by rainfall, groundwater changes, or irrigation. Pattern formation in such systems provides an opportunity to investigate the temporal aspects of spatial self-organization, which have been heretofore understudied. By comparing multiple alternative mechanisms, we found that regular iron banding in upland soils is explained by coupling two sets of scale-dependent feedbacks, the general principle of Turing morphogenesis. First, clay dispersion and coagulation in iron redox fluctuations amplify soil Fe(III) aggregation and crystal growth to a level that negatively affects root growth. Second, the activation of this negative root response to highly crystalline Fe(III) leads to the formation of rhythmic iron bands. In forming iron bands, environmental variability plays a critical role. It creates alternating anoxic and oxic conditions for required pattern-forming processes to occur in distinctly separated times and determines durations of anoxic and oxic episodes, thereby controlling relative rates of processes accompanying oxidation and reduction reactions. As Turing morphogenesis requires ratios of certain process rates to be within a specific range, environmental variability thus modifies the likelihood that pattern formation will occur. Projected changes of climatic regime could significantly alter many spatially self-organized systems, as well as the ecological functioning associated with the striking patterns they present. This temporal dimension of pattern formation merits close attention in the future.


The Utility of Ovotransferrin and Ovotransferrin-Derived Peptides as Possible Candidates in the Clinical Treatment of Cardiovascular Diseases.
Several of the most prevalent etiological factors which contribute towards global death rates are associated with cardiovascular diseases (CVDs), which include a range of conditions such as angina, rheumatic heart disease, and venous thrombosis. Extensive research has been conducted into the role played by oxidative stress and inflammation in the functional transformations associated with the progression of CVDs, while the research findings from these investigations have been both fruitful and informative. In view of the adverse secondary effects that result from the clinical administration of many synthetic medications, research which explored the treatment of severe and long-lasting conditions, including CVDs, has primarily centered on the potential benefits displayed by natural agents, one of which is food protein-based bioactive peptides. Most importantly, previous research has revealed the possible benefits associated with these products' anti-inflammatory and antioxidant characteristics. In light of these considerations, this paper aims to review the degree to which ovotransferrin (otrf, also referred to as conalbumin) and otrf-derived peptides, including IRW, IQW, and KVREGT, are, by virtue of their anti-inflammatory and antioxidant characteristics, viable treatment agents for endothelial dysfunction and the prevention of CVD.


Bites by coral snakes (Micrurus spp.) in Campinas, State of São Paulo, Southeastern Brazil.
Coral snakes (Micrurus spp.) are the main representatives of the Elapidae in South America. However, bites by these snakes are uncommon. We retrospectively reviewed the data from 11 individuals bitten by coral snakes over a 20-year period; four were confirmed (snake brought for identification) and seven were highly suspected (neuromuscular manifestations) cases of elapid envenoming. The cases were classified as dry-bite (n = 1, caused by M. lemniscatus; did not receive antivenom), mild (n = 2, local manifestations with no acute myasthenic syndrome; M. frontalis and Micrurus spp.), moderate (n = 5, mild myasthenia) or severe (n = 3, important myasthenia; one of them caused by M. frontalis). The main clinical features upon admission were paresthesia (local, n = 9; generalized, n = 2), local pain (n = 8), palpebral ptosis (n = 8), weakness (n = 4) and inability to stand up (n = 3). No patient developed respiratory failure. Antivenom was used in ten cases, with mild early reactions occurring in three. An anticholinesterase drug was administered in the three severe cases, with a good response in two. No deaths were observed. Despite the high toxicity of coral snake venoms, the prognosis following envenoming is good. In serious bites by M. frontalis or M. lemniscatus, the venom of which acts postsynaptically, anticholinesterases may be useful as an ancillary measure if antivenom is unavailable, if there is a delay in obtaining a sufficient amount, or in those patients given the highest recommended doses of antivenom without improvement of the paralysis or with delayed recovery.


VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders.
Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.


Do Autistic and Depressed Rats Express the Same Type of Maternal Care?
Autism is a neurodevelopmental disorder that is more frequently diagnosed in men. Nevertheless, through current diagnostic tools, women have also been found to be affected by this disorder, but in different ways. Few studies have been conducted regarding unique periods of life, such as motherhood. Yet, extant literature has already described the existence of a comorbidity between autism and postpartum depression. Thus, this study aimed to compare the maternal care sphere between two animal models of these diseases. Lactating rats were subdivided into three groups (n = 8 animals/group): 1) control dams; 2) maternal separation (MS) dams, separated from their litter for 3 h daily from lactating day (LD) 2-12 for postpartum depression induction; and 3) valproic acid (VPA) dams, which were the pups of dams treated with 400 mg/kg of VPA (i.p.) on gestational day 12.5 for autism induction. Maternal care tests were performed during lactation and, after weaning, dams were euthanized for the analysis of dopaminergic system on the prefrontal cortex. The results showed an impairment of maternal care of MS dams and an improvement of VPA dams, as well as alterations on dopaminergic system that corroborates the behavior data. These findings indicate that VPA dams express better maternal care, even with cognitive and socialization difficulties. This is probably due to a hyper-focus, as opposed to MS dams, which mimic the maternal care dysfunction expressed by women with postpartum depression.


Non-Contact Methods for Measuring Front Cavity Depths of Laboratory Standard Microphones Using a Depth-Measuring Microscope.
To achieve an acceptable degree of accuracy at high frequencies in some standardized methods for primary calibration of laboratory standard (LS) microphones, the front cavity depth lfc of each microphone must be known. This dimension must be measured using non-contact methods to prevent damage to the microphone diaphragm. The basic capabilities of an optical depth-measuring microscope were demonstrated by the agreement of its measurements within 0.7 μm of the known values of reference gage blocks. Using this microscope, two basic methods were applied to measure lfc . One (D) uses direct measurements at the microphone front surface annulus and conventional data reduction techniques. The other (GB) uses measurements at the surface of a gage block placed on the annulus, and plane-fitting data reduction techniques intended to reduce the effects of the slightly imperfect geometries of the microphones. The GB method was developed to provide a smoother surface of measurement than the relatively rough surface of the annulus, and to simulate the contact that occurs between the annulus and the smooth, plane surface of an acoustic coupler during microphone calibration. Using these methods, full data sets were obtained at 33 measurement positions (D), or 25 positions (GB). In addition, D and GB subsampling methods were applied by using subsamples of either the D or the GB full data sets. All these methods were applied to six LS microphones, three each of two different types. The GB subsampling methods are preferred for several reasons. The measurement results for lfc obtained by these methods agree well with those obtained by the GB method using the full data set. The expanded uncertainties of results from the GB subsampling methods are not very different from the expanded uncertainty of results from the GB method using the full data set, and are smaller than the expanded uncertainties of results from the D subsampling methods. Measurements of lfc using the GB subsampling method with only nine measurement positions exhibit expanded uncertainties (with coverage factor k = 2) within 4 μm, and can improve the uncertainty of microphone calibrations by an order of magnitude over the result from use of generic standardized microphone type nominal lfc values and tolerance limits.


Modified error in constitutive equations (MECE) approach for ultrasound elastography.
A partial differential equation-constrained optimization approach is presented for reconstructing mechanical properties (e.g., elastic moduli). The proposed method is based on the minimization of an error in constitutive equations functional augmented with a least squares data misfit term referred to as MECE for "modified error in constitutive equations." The main theme of this paper is to demonstrate several key strengths of the proposed method on experimental data. In addition, some illustrative examples are provided where the proposed method is compared with a common shear wave elastography (SWE) approach. To this end, both synthetic data, generated with transient finite element simulations, as well as ultrasonically tracked displacement data from an acoustic radiation force (ARF) experiment are used in a standard elasticity phantom. The results indicate that the MECE approach can produce accurate shear modulus reconstructions with significantly less bias than SWE.


Stable complex formation of CENP-B with the CENP-A nucleosome.
CENP-A and CENP-B are major components of centromeric chromatin. CENP-A is the histone H3 variant, which forms the centromere-specific nucleosome. CENP-B specifically binds to the CENP-B box DNA sequence on the centromere-specific repetitive DNA. In the present study, we found that the CENP-A nucleosome more stably retains human CENP-B than the H3.1 nucleosome in vitro. Specifically, CENP-B forms a stable complex with the CENP-A nucleosome, when the CENP-B box sequence is located at the proximal edge of the nucleosome. Surprisingly, the CENP-B binding was weaker when the CENP-B box sequence was located in the distal linker region of the nucleosome. This difference in CENP-B binding, depending on the CENP-B box location, was not observed with the H3.1 nucleosome. Consistently, we found that the DNA-binding domain of CENP-B specifically interacted with the CENP-A-H4 complex, but not with the H3.1-H4 complex, in vitro. These results suggested that CENP-B forms a more stable complex with the CENP-A nucleosome through specific interactions with CENP-A, if the CENP-B box is located proximal to the CENP-A nucleosome. Our in vivo assay also revealed that CENP-B binding in the vicinity of the CENP-A nucleosome substantially stabilizes the CENP-A nucleosome on alphoid DNA in human cells.


Immunomodulatory potential of four candidate probiotic Lactobacillus strains from plant and animal origin using comparative genomic analysis.
Probiotic strains from different origins have shown promise in recent decades for their health benefits, for example in promoting and regulating the immune system. The immunomodulatory potential of four Lactobacillus strains from animal and plant origins was evaluated in this paper based on their genomic information. Comparative genomic analysis was performed through genome alignment, average nucleotide identity (ANI) analysis and gene mining for putative immunomodulatory genes. The genomes of the four Lactobacillus strains show relative similarities in multiple regions, as observed in the genome alignment. However, ANI analysis showed that L. mucosae LM1 and L. fermentum SK152 are the most similar when considering their nucleotide sequences alone. Gene mining of putative immunomodulatory genes studied from L. plantarum WCFS1 yielded multiple results in the four potential probiotic strains, with L. plantarum SK151 showing the largest number of genes at around 74 hits, followed by L. johnsonii PF01 at 41 genes when adjusted for matches with at least 30 % identity. Looking at the immunomodulatory genes in each strain, L. plantarum SK151 and L. johnsonii PF01 may have wider activity, covering both immune activation and immune suppression, as compared to L. mucosae LM1 and L. fermentum SK152, which could be more effective in activating immune cells and the pro-inflammatory cascade rather than suppressing it. The similarities and differences between the four Lactobacillus species showed that there is no definitive trend based on the origin of isolation alone. Moreover, higher percentage identities between genomes do not directly correlate with higher similarities in potential activity, such as in immunomodulation. The immunomodulatory function of each of the four Lactobacillus strains should be observed and verified experimentally in the future, since some the activity of some genes may be strain-specific, which would not be identified through comparative genomics alone.


Thimerosal induces apoptotic and fibrotic changes to kidney epithelial cells in vitro.
Thimerosal is an ethyl mercury-containing compound used mainly in vaccines as a bactericide. Although the kidney is a key target for mercury toxicity, thimerosal nephrotoxicity has not received the same attention as other mercury species. The aim of this study was to determine the potential cytotoxic mechanisms of thimerosal on human kidney cells. Human kidney proximal tubular epithelial (HK2) cells were exposed for 24 h to thimerosal (0-2 µM), and assessed for cell viability, apoptosis, and cell proliferation; expression of proteins Bax, nuclear factor-κB subunits, and transforming growth factor-β1 (TGFβ1); mitochondrial health (JC-1, MitoTracker Red CMXRos); and fibronectin levels (enzyme-linked immunosorbent assay). Thimerosal diminished HK2 cell viability and mitosis, promoted apoptosis, impaired the mitochondrial permeability transition, enhanced Bax and TGFβ1 expression, and augmented fibronectin secretion. This is the first report about kidney cell death and pro-fibrotic mechanisms promoted by thimerosal. Collectively, these in vitro results demonstrate that (1) thimerosal induces kidney epithelial cell apoptosis via upregulating Bax and the mitochondrial apoptotic pathway, and (2) thimerosal is a potential pro-fibrotic agent in human kidney cells. We suggest that new evidence on toxicity as well as continuous surveillance in terms of fibrogenesis is required concerning thimerosal use.


The needleless syringe: efficacy of anesthesia and patient preference in child dental patients.
Delivery of local anesthesia in dentistry is primarily by way of a needle-tipped syringe. This procedure, when used on children, is often stressful and sometimes painful for the patient. The Syrijet is an instrument which delivers anesthesia without the use of a needle. This is accomplished by delivering the anesthetic solution under high compressive forces. The subjects were thirty-four children ranging in age between 5 and 15 years, on whom forty-five dental procedures were completed. There was a statistically significant difference in favor of the instrument, with twenty-five subjects reporting a preference for it. The instrument was completely successful in providing anesthesia in thirty-six of the forty-five procedures.


Global varus malalignment increase from double-leg to single-leg stance due to intra-articular changes.
Preoperatively planned correction for tibial osteotomy surgery is usually based on weightbearing long-leg Xrays, while the surgery is performed in a supine non-weightbearing position. The purpose of this study was to assess the differences in lower limb alignment in three different weightbearing conditions: supine position, double-leg (DL) stance and single-sleg (SL) stance prior to performing a medial opening wedge high tibial osteotomy (MOWHTO) for varus malalignment. The hypothesis of this study was that progressive limb-loading would lead to an increased preoperative varus deformity. This retrospective study included 89 patients (96 knees) with isolated medial knee osteoarthritis (Ahlbäck grade I or II) and significant metaphyseal tibial vara (> 6°). The differences between supine position, DL stance and SL stance were analysed for the hip-knee-ankle angle (HKA), lateral distal femoral angle (LDFA), medial proximal tibial angle (MPTA), weight-bearing line ratio (WBL) and joint line convergence angle (JLCA). From a supine position to DL stance, the HKA angle slightly increased from 175.5° ± 1.1° to 176.3° ± 1.1° and JLCA changed from 2.0° ± 0.3° to 1.8° ± 0.3° without a statistically significant difference. From DL to SL stances, the HKA angle decreased from 176.3° ± 1.1° to 174.4° ± 1.1° (p < 0.05) and the JLCA increased from 1.8° ± 0.3° to 2.6° ± 0.3° (p < 0.05). A significant correlation was found between ΔHKA and ΔJLCA between the DL and the SL stances (R2 = 0.46; p = 0.01). Varus malalignment increases with weight-bearing loading from double-leg to single-leg stances with an associated JLCA increase. Thus, single-leg stance radiographs may be useful to correct preoperative planning considering patient-specific changes in JLCA. III, retrospective comparative study.


Evaluation of a sun safety education programme for primary school students in Switzerland.
The incidence of skin cancer has increased worldwide, with rates being especially high in Switzerland compared with other European countries. Extensive sun exposure during childhood is considered a key factor for skin carcinogenesis. The aim of the study was to evaluate the impact of a school-based sun safety education programme developed by the Swiss Cancer Leagues on primary school students' sun-related knowledge, protective behaviours and sunburn rates. In summer 2011, 1-h sun safety education sessions were held at 33 primary schools throughout the Canton of Zurich (North-Eastern Switzerland). Children in the participating school classes (first, second and third graders) answered a questionnaire on their sun-related knowledge, behaviours and sunburn experience shortly before and 1 year after the intervention. Overall, 3110 completed pretest and 1738 post-test questionnaires were eligible for analysis. The evaluation of pretest data showed considerable room for improvement in terms of sun-related knowledge, considering that merely a good half of the children were conscious that the sun may present a hazard to health. Overall, more than 95% of students benefited from the protection of sunscreen (application by parents: 73%; application by child: 66%), but only 36% stated that they generally sought shade on sunny days. After the intervention, knowledge increased considerably and significantly (P<0.0001), but there was no change in sun-protective behaviours (use of sunscreen, seeking shade). However, we observed a nonsignificant trend towards decreased sunburn rates. The brief one-time sun safety education sessions were effective in sustainably improving children's sun-related knowledge and possibly to some extent in decreasing their sunburn rates.


Effects of calcium on renin and aldosterone.
A series of experiments was undertaken to assess the effects of calcium administration, in vivo, on renin and aldosterone secretion. In the anesthetized dog, renin secretion was decreased by renal arterial infusions of calcium chloride and calcium gluconate; aldosterone excretion was not affected. In the sodium chloride-deprived rat, dietary calcium chloride loading decreased plasma renin activity, whereas calcium gluconate did not. Both calcium salts increased aldosterone production. In the non-filtering, denervated, papaverine-treated dog kidney, renin release was stimulated by renal arterial infusion of verapamil. In the rat, chronic oral verapamil administration decreased plasma aldosterone but had no effect on renin. In humans, chronic oral verapamil decreased aldosterone responsiveness to infusion of angiotensin II. Thus, in vivo renin release is inhibited by hypercalcemia and stimulated by blocking calcium transport; conversely, aldosterone production is stimulated by a high calcium intake and inhibited by blocking calcium transport. These effects of calcium on renin and aldosterone may have implications for understanding the putative relation between calcium and hypertension.


Early Life Experiences Moderate the Relationship Between Genetic Risk of Autism and Current and Lifetime Mental Health.
Although individuals with autism are at greater risk of mental health challenges than others, we know little about the relationship between the mental health of older adults (50+) and autism because they are less likely to be diagnosed. Identifying the risk and protective factors that are associated with mental health can increase educational awareness, inform clinical practice, and provide information to help diagnose and treat older adults with autism. This study used longitudinal panel data of the 2008-2016 waves of the Health and Retirement Study. It estimated individual random-effect models by interacting a genetic propensity toward autism and early life experiences to test whether the latter has a moderating effect on the relationships between genetics and the Center for Epidemiologic Studies Depression (CES-D) score, self-reported depression, and history of psychiatric problems. Results suggest that individuals with a higher genetic propensity for autism are less likely to develop psychiatric problems if they report a positive maternal relationship early in life. Further, a combined effect of police encounters early in life and genetic risk for autism is associated with higher CES-D scores, increased odds of self-reported depression, and a history of psychiatric problems. Clinical applications of these findings include the need to establish and support high-quality relationships by addressing both child and caregiver needs. Further, these findings support the need to design and implement proactive interventions to teach police and autistic individuals how to successfully navigate these encounters.


Membership function modification of fuzzy logic controllers with histogram equalization.
In most fuzzy logic controllers (FLCs), initial membership functions (MFs) are normally laid evenly all across the universes of discourse (UD) that represent fuzzy control inputs. However, for evenly distributed MFs, there exists a potential problem that may adversely affect the control performance; that is, if the actual inputs are not equally distributed, but instead concentrate within a certain interval that is only part of the entire input area, this will result in two negative effects. On one hand, the MFs staying in the dense-input area will not be sufficient to react precisely to the inputs, because these inputs are too close to each other compared to the MFs in this area. The same fuzzy control output could be triggered for several different inputs. On the other hand, some of the MFs assigned for the sparse-input area are "wasted". In this paper we argue that, if we arrange the placement of these MFs according to a statistical study of feedback errors in a closed-loop system, we can expect a better control performance. To this end, we introduce a new mechanism to modify the evenly distributed MFs with the help of a technique termed histogram equalization. The histogram of the errors is actually the spatial distribution of real-time errors of the control system. To illustrate the proposed MF modification approach, a computer simulation of a simple system that has a known mathematical model is first analyzed, leading to our understanding of how this histogram-based modification mechanism functions. We then apply this method to an experimental laser tracking system to demonstrate that in real-world applications, a better control performance can he obtained by using this proposed technique.


Ceruloplasmin is associated with the infiltration of immune cells and acts as a prognostic biomarker in patients suffering from glioma.
Glioma is regarded as a prevalent form of cancer that affects the Central Nervous System (CNS), with an aggressive growth pattern and a low clinical cure rate. Despite the advancement of the treatment strategy of surgical resection, chemoradiotherapy and immunotherapy in the last decade, the clinical outcome is still grim, which is ascribed to the low immunogenicity and tumor microenvironment (TME) of glioma. The multifunctional molecule, called ceruloplasmin (CP) is involved in iron metabolism. Its expression pattern, prognostic significance, and association with the immune cells in gliomas have not been thoroughly investigated. Studies using a variety of databases, including Chinese Glioma Genome Atlas (CGGA), The Cancer Genome Atlas (TCGA), and Gliovis, showed that the mRNA and protein expression levels of CP in patients suffering from glioma increased significantly with an increasing glioma grade. Kaplan-Meier (KM) curves and statistical tests highlighted a significant reduction in survival time of patients with elevated CP expression levels. According to Cox regression analysis, CP can be utilized as a stand-alone predictive biomarker in patients suffering from glioma. A significant association between CP expression and numerous immune-related pathways was found after analyzing the data using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). Tumor Immune Estimation Resource (TIMER) and CIBERSORT analyses indicated a substantial correlation between the CP expression and infiltration of immunocytes in the TME. Additionally, immune checkpoints and CP expression in gliomas showed a favorable correlation. According to these results, patients with glioma have better prognoses and levels of tumor immune cell infiltration when their CP expression is low. As a result, CP could be used as a probable therapeutic target for gliomas and potentially anticipate the effectiveness of immunotherapy.


MicroRNA‑766 inhibits the malignant biological behaviours of pancreatic ductal adenocarcinoma by directly targeting ETS1.
Increasing evidence indicates that numerous microRNAs (miRNAs) are altered in pancreatic ductal adenocarcinoma (PDAC), and their alterations significantly influence the malignant behaviour of PDAC. Therefore, identifying miRNAs associated with PDAC and their biological roles in the disease may provide promising therapeutic opportunities. Alteration of the expression of miRNA‑766 (miR‑766) has been previously reported in several types of human malignancy. However, to the best of our knowledge, whether miR‑766 exhibits different expression patterns in PDAC and its underlying functions in the progression of PDAC remain to be elucidated. In the present study, reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was used to detect miR‑766 expression levels in PDAC tissues and cell lines. The effects of miR‑766 upregulation on PDAC cell proliferation and invasion were evaluated using MTT and invasion assays, respectively. The mechanisms underlying the role of miR‑766 in PDAC cells were explored using bioinformatics analysis, luciferase reporter assay, RT‑qPCR and western blot analysis. It was found that miR‑766 was significantly downregulated in PDAC tissues and cell lines. The detailed roles of miR‑766 in the progression of PDAC were characterised using Panc‑1 and Aspc‑1 cell lines. The results revealed that the upregulation of miR‑766 restricted the proliferation and invasion of PDAC cells. Through a series of experiments, it was found that E26 transformation specific‑1 (ETS1) was a direct target of miR‑766 in PDAC cells. Furthermore, ETS1 knockdown simulated the inhibitory effects of the overexpression of miR‑766 on PDAC cells, whereas the effects of miR‑766 restoration on the PDAC cells were reversed by overexpressing ETS1. In conclusion, the findings of the present study demonstrate that miR‑766 offers potential as a therapeutic target for patients with PDAC.


Chemically versus thermally processed brown shrimp shells or Chinese mitten crab as a source of chitin, nutrients or salts and as microbial stimulant in soilless strawberry cultivation.
Brown shrimp (Crangon crangon) shells and Chinese mitten crab (Eriocheir sinensis) were chemically demineralized and deproteinized (denoted as M1 to M4 for the shrimp shells and M5 to M7 for the Chinese mitten crab), and shrimp shells were torrefied at 200 to 300 °C (denoted as R200, R255, R300), and were compared with a commercially available chitin source (denoted as reference chitin). Based on their chemical characteristics, a selection of chitin sources was tested for their N mineralization capacity. The N release was high for the chemically treated shrimp shells and Chinese mitten crab, but not for the torrefied shrimp shells with or without acid treatment, indicating that treatment at 200 °C or higher resulted in low N availability. Interaction with nutrients was tested in a leaching experiment with limed peat for three thermally and two chemically processed shrimp shells and the reference chitin source. The K concentrations in the leachate for the chemically treated shrimp shells and the reference chitin were lower than for limed peat during fertigation. Irreversible K retention was observed for one source of chemically treated shrimp shells, and the reference chitin. The thermally treated shrimp shells had a significantly higher net release of P, Na and Cl than the treatment without chitin source. Three shrimp shell based materials (M4, R200 and R300) and the reference chitin were tested in a greenhouse trial with strawberry at a dose of 2 g/L limed peat. A very positive and significant effect on Botrytis cinerea disease suppression in the leaves was found for the reference chitin, M4 and R200 compared to the unamended control. The disease suppression of the 3 chitin sources was linked with an increase of the microbial biomass in the limed peat with 24% to 28% due to chitin decomposition and a 9-44% higher N uptake in the plants.


RNA fingerprinting displays UVB-specific disruption of transcriptional control in human melanocytes.
In mammalian cells, UV induces a limited set of early transcribed genes, which overlaps with the set of genes induced by tumor promoting drugs such as 12-O-tetradecanoyl phorbol-13-acetate (TPA). Among these are genes for transcription factors, the activation of which leads to complex secondary changes in expression of multiple target genes. How these delayed pleiotropic UV effects on transcription may contribute to initiation of melanoma skin cancer is poorly understood. We analyzed changes in the relative abundances of 1900 transcripts in newborn human melanocytes 8 h after treatment with UVB, TPA, and cycloheximide in all combinations, using RNA arbitrarily primed PCR for differential display. The relative abundances of 205 transcripts (11% of all transcripts surveyed) were altered by one or more of the treatment combinations. Fourteen of the 77 genes up-regulated by TPA were also up-regulated by UVB, but 60 of the TPA up-regulated genes were down-regulated by UVB, indicating both intersecting and independent signal transduction pathways for UVB and TPA. A number of UVB and TPA target genes were identified by cDNA cloning. Consistent with UVB induction of a partly transformed phenotype in mammalian cells, UVB antagonized the TPA-inducible expression of tumor-suppressive tropomyosin 3 mRNA. In addition, UVB may impair mitochondrial functioning and induce oxidative stress by strong down-regulation of mitochondrial transcription. Finally, increased expression of the dihydropteridine reductase gene, a major regulator of the cellular tetrahydrobiopterin pool, was linked to the UV pathway.


Enhanced intracellular accumulation and cytotoxicity of bortezomib against liver cancer cells using N-stearyl lactobionamide surface modified solid lipid nanoparticles.
Asialoglycoprotein receptors (ASGPRs) are highly expressed on hepatocytes and have been used for liver-targeted delivery and hepatocellular carcinoma (HCC) therapy. However, targeted delivery of bortezomib (BTZ) to HCC has not been reported. In this study, N-stearyl lactobionamide (N-SALB) with galactose (Gal) moiety was synthesized as a targeting agent and its structure was confirmed by FT-IR and NMR analyses. N-SALB surface-modified solid lipid nanoparticles (SLNs) loaded with BTZ (Gal-SLNs/BTZ) were developed to target BTZ delivery into HCC cancer cells. The Gal-SLNs/BTZ had an average particle size of 116.3 nm, polydispersity index (PDI) of 0.210, and zeta potential of -13.8 mV. TEM analysis showed their nanometer-sized spherical morphology. The encapsulation efficiency (EE) and drug loading (DL) capacity were 84.5 % and 1.16 %, respectively. Release studies showed that BTZ loaded inside the SLNs was slowly released over a period of 72 h at pH 7.4. Flow cytometry analysis showed significantly higher intracellular uptake of N-SALB-targeted nanoparticles than non-targeted nanoparticles in HepG2 cells. All lipid formulations showed good biocompatibility in the cytotoxicity study using MTT assay. Concentration-dependent cytotoxicity was observed for all formulations, with N-SALB-targeted nanoparticles demonstrating more cytotoxicity against HepG2 cells. The highest percentage of apoptosis was obtained for N-SALB-targeted nanoparticles compared to non-targeted nanoparticles (42.2 % and 8.70 %, respectively). Finally, biodistribution studies in HepG2 bearing nude mice showed that the accumulation of targeted nanoparticles in the tumor was significantly higher than non-targeted nanoparticles.


Regulation of toll-like receptor signaling by the SF3a mRNA splicing complex.
The innate immune response plays a key role in fighting infection by activating inflammation and stimulating the adaptive immune response. However, chronic activation of innate immunity can contribute to the pathogenesis of many diseases with an inflammatory component. Thus, various negatively acting factors turn off innate immunity subsequent to its activation to ensure that inflammation is self-limiting and to prevent inflammatory disease. These negatively acting pathways include the production of inhibitory acting alternate proteins encoded by alternative mRNA splice forms of genes in Toll-like receptor (TLR) signaling pathways. We previously found that the SF3a mRNA splicing complex was required for a robust innate immune response; SF3a acts to promote inflammation in part by inhibiting the production of a negatively acting splice form of the TLR signaling adaptor MyD88. Here we inhibit SF3a1 using RNAi and subsequently perform an RNAseq study to identify the full complement of genes and splicing events regulated by SF3a in murine macrophages. Surprisingly, in macrophages, SF3a has significant preference for mRNA splicing events within innate immune signaling pathways compared with other biological pathways, thereby affecting the splicing of specific genes in the TLR signaling pathway to modulate the innate immune response.


Simultaneous acquisition of complex transmittance and birefringence with two counter-rotating, circularly polarized THz pulses.
Here we demonstrate simultaneous measurements of the complex transmittance and birefringence using left- and right-handed circularly polarized terahertz (THz) pulses. We change the polarization of the THz pulses periodically by modulating the polarization of the pump pulses directed onto a ZnTe (111) crystal, and we convert linear to circular polarization using a broadband THz quarter-wave retarder. By integrating the alternating-emission system with the polarization-sensitive terahertz time-domain spectrometer, we are able to obtain the electric-field vector of the transmitted terahertz pulses for both the left- and right-handed circular polarizations. Utilizing this technique, we are able to measure simultaneously the frequency-dependent complex refractive indices (real and imaginary parts) and the orientations of the slow and fast axes of birefringent materials, a quartz disc and a barium borate crystal, in a single temporal sweep.


Predictive factors for response to abiraterone in metastatic castration refractory prostate cancer.
Management of metastatic castration refractory prostate cancer (CRPC) is rapidly evolving. Rationalisation of treatment requires identification of those patients more likely to benefit from a particular therapy. We reviewed the outcome of patients treated with abiraterone at our Institution to describe factors predictive for response. Patients with CRCP treated with abiraterone were identified. Baseline variables and potential prognostic factors were extracted from electronic records. Outcome measures included overall survival (OS), prostate-specific antigen (PSA) response and time to PSA progression (TTPP). The Kaplan-Meier method and Cox proportional hazards model were used to analyze survival data. A total of 61 patients met the inclusion criteria. In multivariate analysis, three independent predictors of OS were identified: Duration of response to androgen deprivation therapy (ADT) (hazard ratio(HR)=0.95, p=0.006), performance status (HR=7.4, p=0.013), and baseline haemoglobin (HR=0.47, p≤0.001). This study has identified three factors predictive for response to abiraterone in CRPC. Duration of response to ADT has not been previously shown to be a predictive factor for patients with CRCP. We suggest that a prospective validation is required.


Quantifying Intranasally Administered Deferoxamine in Rat Brain Tissue with Mass Spectrometry.
Deferoxamine, a metal chelator, has been shown to be neuroprotective in animal models of ischemic stroke, traumatic brain injury and both subarachnoid and intracerebral hemorrhage. Intranasal deferoxamine (IN DFO) has also shown promise as a potential treatment for multiple neurodegenerative diseases, including Parkinson's and Alzheimer's. However, there have been no attempts to thoroughly understand the dynamics and pharmacokinetics of IN DFO. We developed a new high-performance liquid-chromatography electrospray-tandem mass spectrometry (HPLC/ESI-MS2) method to quantify the combined total levels of DFO, ferrioxamine (FO; DFO bound to iron), and aluminoxamine (AO; aluminum-bound DFO) in brain tissue using a custom-synthesized deuterated analogue (DFO-d7, Medical Isotopes Inc., Pelham NH) as an internal standard. We applied our method toward understanding the pharmacokinetics of IN DFO delivery to the brain and blood of rats from 15 min to 4 h after delivery. We found that IN delivery successfully targets DFO to the brain to achieve concentrations of 0.5-15 μM in various brain regions within 15 min, and decreasing though still detectable after 4 h. Systemic exposure was minimized as assessed by concentration in blood serum. Serum concentrations were 0.02 μM at 15 min and no more than 0.1 μM at later time points. Compared to blood serum, brain region-specific drug exposure (as measured by area under the curve) ranged from slightly under 10 times exposure in the hippocampus to almost 200 times exposure in the olfactory bulb with IN DFO delivery. These findings represent a major step toward future method development, pharmacokinetic studies, and clinical trials for this promising therapeutic.


[DPC data based situation analysis of Regional Core Hospital: an application of the GIS Methodology for Regional Health Care Plans].
Each prefectural government must formulate a Regional Health Care Plan (RHCP) which will be enforced from 2013 to 2017. During this period, in Health Care Regions (HCRs) where the population is less than 200,000, the inflow rate of inpatients is less than 20%, and the outflow rate of inpatients is more than 20%, each prefecture is required to reconsider the size of any such secondary HCR. They should consider natural conditions such as geographical characteristics, social conditions such as traffic, patient flow of each area, access time to core hospitals, et cetera. In this study, we analyzed the situation of the Kiso secondary HCR of Nagano prefecture, based on the 2010 Diagnosis Procedure Combination (DPC) data that was distributed by the Ministry of Health, Labour and Welfare. According to a Geographic Information System (GIS) analysis, the Kiso HCR is self-sufficient for most emergency care and cancer treatments, even though this area is classified as a rural HCR. As this research indicates, DPC data is a useful resource for HCR planning.


Fertility preservation discussions, referral and follow-up in male-to-female and female-to-male adolescent transgender patients.
The number of patients seeking transgender healthcare is growing, and there is a potential impact of gender-affirming therapies on fertility. The use of fertility preservation (FP), particularly among transgender adolescents, has been limited. We aimed to examine differences in FP counselling, referral and utilisation between male-to-female (MtF) and female-to-male (FtM) transgender adolescents. A retrospective review of the medical records of patients ages 12-17 seen at an academic medical centre between 2012 and 2017 with a diagnosis of gender dysphoria was conducted. A total of 22 MtF and 45 FtM adolescents were included. The counselling on the potential fertility impact of gender-affirming therapy was documented in 55%, and of those counselled, 73% were counselled before receiving medication. There was no significant difference between the timing of counselling for MtF versus FtM adolescents. Of patients with documented reproductive wishes, 77% reported either desire for adopted children or no desire for biological children. Among patients offered FP referral, 2 (22.2%) MtF and 3 (12.5%) FtM patients accepted; both MtF patients cryopreserved sperm. While most adolescents were counselled on the fertility impact of gender-affirming therapy, there is room for improvement as 45% of patients had no documented counselling. The rate of transgender adolescents pursuing FP consultation and gamete cryopreservation was low, consistent with prior studies in this population.


Transcriptional Homeostasis of Oxidative Stress-Related Pathways in Altered Gravity.
Whereby several types of cultured cells are sensitive to gravity, the immune system belongs to the most affected systems during spaceflight. Since reactive oxygen species/reactive nitrogen species (ROS/RNS) are serving as signals of cellular homeostasis, particularly in the cells of the immune system, we investigated the immediate effect of altered gravity on the transcription of 86 genes involved in reactive oxygen species metabolism, antioxidative systems, and cellular response to oxidative stress, using parabolic flight and suborbital ballistic rocket experiments and microarray analysis. In human myelomonocytic U937 cells, we detected a rapid response of 19.8% of all of the investigated oxidative stress-related transcripts to 1.8 g of hypergravity and 1.1% to microgravity as early as after 20 s. Nearly all (97.2%) of the initially altered transcripts adapted after 75 s of hypergravity (max. 13.5 g), and 100% adapted after 5 min of microgravity. After the almost complete adaptation of initially altered transcripts, a significant second pool of differentially expressed transcripts appeared. In contrast, we detected nearly no response of oxidative stress-related transcripts in human Jurkat T cells to altered gravity. In conclusion, we assume a very well-regulated homeostasis and transcriptional stability of oxidative stress-related pathways in altered gravity in cells of the human immune system.


BIOB: a mathematical model for the biodegradation of low solubility hydrocarbons.
Modeling oil biodegradation is an important step in predicting the long term fate of oil on beaches. Unfortunately, existing models do not account mechanistically for environmental factors, such as pore water nutrient concentration, affecting oil biodegradation, rather in an empirical way. We present herein a numerical model, BIOB, to simulate the biodegradation of insoluble attached hydrocarbon. The model was used to simulate an experimental oil spill on a sand beach. The biodegradation kinetic parameters were estimated by fitting the model to the experimental data of alkanes and aromatics. It was found that parameter values are comparable to their counterparts for the biodegradation of dissolved organic matter. The biodegradation of aromatics was highly affected by the decay of aromatic biomass, probably due to its low growth rate. Numerical simulations revealed that the biodegradation rate increases by 3-4 folds when the nutrient concentration is increased from 0.2 to 2.0 mg N/L.


Integrating natural history collections and comparative genomics to study the genetic architecture of convergent evolution.
Evolutionary convergence has been long considered primary evidence of adaptation driven by natural selection and provides opportunities to explore evolutionary repeatability and predictability. In recent years, there has been increased interest in exploring the genetic mechanisms underlying convergent evolution, in part, owing to the advent of genomic techniques. However, the current 'genomics gold rush' in studies of convergence has overshadowed the reality that most trait classifications are quite broadly defined, resulting in incomplete or potentially biased interpretations of results. Genomic studies of convergence would be greatly improved by integrating deep 'vertical', natural history knowledge with 'horizontal' knowledge focusing on the breadth of taxonomic diversity. Natural history collections have and continue to be best positioned for increasing our comprehensive understanding of phenotypic diversity, with modern practices of digitization and databasing of morphological traits providing exciting improvements in our ability to evaluate the degree of morphological convergence. Combining more detailed phenotypic data with the well-established field of genomics will enable scientists to make progress on an important goal in biology: to understand the degree to which genetic or molecular convergence is associated with phenotypic convergence. Although the fields of comparative biology or comparative genomics alone can separately reveal important insights into convergent evolution, here we suggest that the synergistic and complementary roles of natural history collection-derived phenomic data and comparative genomics methods can be particularly powerful in together elucidating the genomic basis of convergent evolution among higher taxa. This article is part of the theme issue 'Convergent evolution in the genomics era: new insights and directions'.


Prototyping Apps for the Management of Sleep, Fatigue, and Behavioral Health in Austere Far-Forward Environments: Development Study.
Military service inherently includes frequent periods of high-stress training, operational tempo, and sustained deployments to austere far-forward environments. These occupational requirements can contribute to acute and chronic sleep disruption, fatigue, and behavioral health challenges related to acute and chronic stress and disruption of team dynamics. To date, there is no centralized mobile health platform that supports self- and supervised detection, monitoring, and management of sleep and behavioral health issues in garrison and during and after deployments. The objective of this study was to adapt a clinical decision support platform for use outside clinical settings, in garrison, and during field exercises by medics and soldiers to monitor and manage sleep and behavioral health in operational settings. To adapt an existing clinical decision support digital health platform, we first gathered system, content, and context-related requirements for a sleep and behavioral health management system from experts. Sleep and behavioral health assessments were then adapted for prospective digital data capture. Evidence-based and operationally relevant educational and interventional modules were formatted for digital delivery. These modules addressed the management and mitigation of sleep, circadian challenges, fatigue, stress responses, and team communication. Connectivity protocols were adapted to accommodate the absence of cellular or Wi-Fi access in deployed settings. The resulting apps were then tested in garrison and during 2 separate field exercises. Based on identified requirements, 2 Android smartphone apps were adapted for self-monitoring and management for soldiers (Soldier app) and team supervision and intervention by medics (Medic app). A total of 246 soldiers, including 28 medics, received training on how to use the apps. Both apps function as expected under conditions of limited connectivity during field exercises. Areas for future technology enhancement were also identified. We demonstrated the feasibility of adapting a clinical decision support platform into Android smartphone-based apps to collect, save, and synthesize sleep and behavioral health data, as well as share data using adaptive data transfer protocols when Wi-Fi or cellular data are unavailable. The AIRE (Autonomous Connectivity Independent System for Remote Environments) prototype offers a novel self-management and supervised tool to augment capabilities for prospective monitoring, detection, and intervention for emerging sleep, fatigue, and behavioral health issues that are common in military and nonmilitary high-tempo occupations (eg, submarines, long-haul flights, space stations, and oil rigs) where medical expertise is limited.


An efficient on-board metal-free nanocatalyst for controlled room temperature hydrogen production.
Positively charged functionalized carbon nanodots (CNDs) with a variety of different effective surface areas (ESAs) are synthesized via a cheap and time effective microwave method and applied for the generation of hydrogen via hydrolysis of sodium borohydride. To the best of our knowledge, this is the first report of metal-free controlled hydrogen generation. Our observation is that a positively charged functional group is essential for the hydrolysis for hydrogen production, but the overall activity is found to be enhanced with the ESA. A maximum value of 1066 ml g-1 min-1 as the turnover frequency is obtained which is moderate in comparison to other catalysts. However, the optimum activation energy is found to be 22.01 kJ mol-1 which is comparable to well-known high cost materials like Pt and Ru. All of the samples showed good reusability and 100% conversion even after the 10th cycle. The effect of H+ and OH- is also studied to control the on-board and on-demand hydrogen production ("on-off switching"). It is observed that H2 production decreases inversely with NaOH concentration and ceases completely when 10-1 M NaOH is added. With the addition of HCl, H2 production can be initiated again, which confirms the on/off control over production.


Novel silicone stent to treat tracheobronchial lesions: results of 35 patients.
We describe a case series of 35 patients with either benign (14) or malignant (21) tracheal stenosis who were treated using a novel silicone stent, the HCPA-1, designed to prevent migration. Between March 2001 and September 2008, 13 women and 22 men received 41 HCPA-1 stents. The median duration of stenting in benign cases was 457 days (range, 4-2,961 days). Successful stent removal with curative results was accomplished in 2 patients with tracheomalacia and 1 with post-intubation stenosis. In malignant cases, the median duration of stenting was 162 days (range, 1-1,279 days). Five patients had tumor progression with obstruction requiring repeated laser resection, dilatation, or additional stents. Two patients died due to airway obstruction despite bronchoscopic intervention. Twelve patients with malignant lesions died with the stent in place. At the end of the study, 3 patients with malignant disease remained alive; 2 were lost to follow-up. The HCPA-1 stent proved to be safe, with no severe complications during the study period, and effective in improving quality of life with relief of dyspnea.


Preclinical activity of multiple-target gold(III)-dithiocarbamato peptidomimetics in prostate cancer cells and xenografts.
Recently, novel gold(III)-dithiocarbamato peptidomimetics, designed to target peptide transporters upregulated in several tumor cells have shown promise as anticancer agents. The biological behavior of the most promising derivatives AuD8 and AuD9 was studied in PC3 and DU145 prostate cancer cells. They exert higher cytotoxicity in vitro than the reference drug cisplatin and induce apoptosis, promoting mitochondrial membrane permeabilization and stimulating reactive oxygen species generation. Moreover, they inhibit both selenoenzyme thioredoxin reductase and proteasome activity. Additionally, AuD8 effectively reduces tumor growth in prostate tumor-bearing nude mice with minimal systemic toxicity. Altogether, our results provide insights into the anticancer activity of these gold(III)-dithiocarbamato peptidomimetics and support their potential as new agents for prostate cancer treatment.