Structural insights into inhibition of lipid I production in bacterial cell wall synthesis.
Antibiotic-resistant bacterial infection is a serious threat to public health. Peptidoglycan biosynthesis is a well-established target for antibiotic development. MraY (phospho-MurNAc-pentapeptide translocase) catalyses the first and an essential membrane step of peptidoglycan biosynthesis. It is considered a very promising target for the development of new antibiotics, as many naturally occurring nucleoside inhibitors with antibacterial activity target this enzyme. However, antibiotics targeting MraY have not been developed for clinical use, mainly owing to a lack of structural insight into inhibition of this enzyme. Here we present the crystal structure of MraY from Aquifex aeolicus (MraYAA) in complex with its naturally occurring inhibitor, muraymycin D2 (MD2). We show that after binding MD2, MraYAA undergoes remarkably large conformational rearrangements near the active site, which lead to the formation of a nucleoside-binding pocket and a peptide-binding site. MD2 binds the nucleoside-binding pocket like a two-pronged plug inserting into a socket. Further interactions it makes in the adjacent peptide-binding site anchor MD2 to and enhance its affinity for MraYAA. Surprisingly, MD2 does not interact with three acidic residues or the Mg(2+) cofactor required for catalysis, suggesting that MD2 binds to MraYAA in a manner that overlaps with, but is distinct from, its natural substrate, UDP-MurNAc-pentapeptide. We have determined the principles of MD2 binding to MraYAA, including how it avoids the need for pyrophosphate and sugar moieties, which are essential features for substrate binding. The conformational plasticity of MraY could be the reason that it is the target of many structurally distinct inhibitors. These findings can inform the design of new inhibitors targeting MraY as well as its paralogues, WecA and TarO.


Descending aortography with balloon inflation. A technique for evaluating the size of persistent ductus arteriosus in infants with large proximal left to right shunts.
Six infants underwent angiography of the aortic root and the descending aorta with balloon inflation during diagnostic cardiac catheterisation and angiography. In these infants with large proximal left to right shunts, descending aortography with balloon inflation resulted in better opacification of the persistent ductus arteriosus than aortic root angiography, thereby enabling accurate measurement of the persistent ductus arteriosus. Similar systemic and pulmonary vascular impedances had been speculated as the cause of poor or non-opacification of the ductus after ascending aortography. Balloon inflation may have altered the aortic impedance with resulting opacification of the ductus. Descending aortic angiography with balloon inflation is also useful in opacifying the isolated ductus and in demonstrating aortopulmonary collaterals in patients with pulmonary atresia.


Molecular identification and immunological characteristics of goose suppressor of cytokine signaling 1 (SOCS-1) in vitro and vivo following DTMUV challenge.
Purpose suppressor of cytokine signaling 1 (SOCS-1) is inducible feedback inhibitors of cytokine signaling and involved in viral infection through regulation of both innate and adaptive immunity. In this study, we firstly cloned SOCS-1 (goSOCS-1) from duck Tembusu virus (DTMUV) infected goose. The full-length sequence of goSOCS-1 ORF is 624bp and encoded 108 amino acids. Structurally, the mainly functional regions (KIR, SH2, SOCS-box) were conserved between avian and mammalian. The tissues distribution data showed SOCS-1 highly expressed in immune related tissues (SP, LU, HG) of both gosling and adult goose. Moreover, the goSOCS-1 transcripts were induced by goIFNs in GEFs and by TLR ligands in PBMCs. Notably, upon DTMUV infection, highly expression level of goSOCS-1 was detected in vitro and in vivo with high viral load. Our results indicated that goSOCS-1 might involve in both innate and adaptive antiviral immunity of waterfowl.


Feasibility and acceptability of comprehensive virtual treatment in eating disorders: perspectives from patients, parents and therapists during the COVID-19 pandemic.
Lockdowns and social distancing as a result of the COVID-19 pandemic have brought about the need to continue treatment virtually in patients with Eating Disorders (ED). To evaluate feasibility, acceptability and adherence to virtual treatment in patients, families and therapists. Fourteen patients, 10 family members and eight therapists from an intensive outpatient program for ED answered online surveys and a SWOT analysis was performed with the responses. Virtual treatment during lockdown was considered feasible and useful by all respondents. Fear of contagion and the presence of parents in the home were identified as strengths. Parents reported problems with nutritional plan compliance, especially in anorexia patients. Therapists highlighted the importance of methodological adaptations in sessions to improve participation. Adherence to sessions was 100% for family members and 90% for patients. Adaptation to a virtual program is a valid and useful option during lockdowns. It improves family participation, but does not replace face-to-face treatment.


Alkaloids from Hippeastrum papilio.
Galanthamine, an acetylcholinesterase inhibitor marketed as a hydrobromide salt (Razadyne®, Reminyl®) for the treatment of Alzheimer's disease (AD), is obtained from Amaryllidaceae plants, especially those belonging to the genera Leucojum, Narcissus, Lycoris and Ungernia. The growing demand for galanthamine has prompted searches for new sources of this compound, as well as other bioactive alkaloids for the treatment of AD. In this paper we report the isolation of the new alkaloid 11β-hydroxygalanthamine, an epimer of the previously isolated alkaloid habranthine, which was identified using NMR techniques. It has been shown that 11β-hydroxygalanthamine has an important in vitro acetylcholinesterase inhibitory activity. Additionally, Hippeastrum papilio yielded substantial quantities of galanthamine.


Phytophthora sojae effector PsCRN70 suppresses plant defenses in Nicotiana benthamiana.
Phytophthora sojae, an oomycete pathogen, produces a large number of effector proteins that enter into host cells. The Crinklers (Crinkling and Necrosis, CRN) are cytoplasmic effectors that are conserved in oomycete pathogens and their encoding genes are highly expressed at the infective stages in P. sojae. However, their roles in pathogenesis are largely unknown. Here, we functionally characterized an effector PsCRN70 by transiently and stably overexpressing it in Nicotiana benthamiana. We demonstrated that PsCRN70 was localized to the plant cell nucleus and suppressed cell death elicited by all the tested cell death-inducing proteins, including BAX, PsAvh241, PsCRN63, PsojNIP and R3a/Avr3a. Overexpression of the PsCRN70 gene in N. benthamiana enhanced susceptibility to P. parasitica. The H2O2 accumulation in the PsCRN70-transgenic plants was reduced compared to the GFP-lines. The transcriptional levels of the defense-associated genes, including PR1b, PR2b, ERF1 and LOX, were also down-regulated in the PsCRN70-transgenic lines. Our results suggest that PsCRN70 may function as a universal suppressor of the cell death induced by many elicitors, the host H2O2 accumulation and the expression of defense-associated genes, and therefore promotes pathogen infection.


Investigating the ways in which health information technology can promote antimicrobial stewardship: a conceptual overview.
Antimicrobial resistance is now recognised as a threat to health worldwide. Antimicrobial stewardship aims to promote the responsible use of antibiotics and is high on international and national policy agendas. Health information technology has the potential to support antimicrobial stewardship in a number of ways, but this field is still poorly characterised and understood. Building on a recent systematic review and expert roundtable discussions, we take a lifecycle perspective of antibiotic use in hospitals and identify potential targets for health information technology-based interventions to support antimicrobial stewardship. We aim for this work to help chart a future research agenda in this critically important area.


Benzo[cd]triangulene: A Spin 1/2 Graphene Fragment.
How does edge modification affect spin distribution in open-shell graphene fragments? We investigated this effect by analyzing spin-delocalization in benzo[cd]-triangulene, a spin 1/2 graphene fragment composed of seven benzenoid rings fused in a hybrid zigzag/armchair fashion. Six rings of this system form the core of Clar's hydrocarbon triangulene, to which an additional ring is annulated in the zigzag region. The singly occupied molecular orbital (SOMO) of this hydrocarbon radical resembles both SOMOs of triangulene, but the spin density is distributed over the core in a nonuniform fashion. The uneven spin distribution is reflected in the reactivity-reaction with oxygen occurs selectively at a position with the highest spin density-and correlates nicely with relative stabilities of the corresponding Clar resonance structures. The spin distribution is different from that of a topologically similar compound composed of the same number of sp2 carbon atoms but featuring six rings only, illustrating the impact of subtle structural changes on spin-density distribution. This compound was characterized by means of UV-vis and electron paramagnetic resonance spectroscopy, cyclic voltammetry, mass spectrometry, and X-ray crystallography. The experimental results are supported by density functional theory calculations.


Cardiac sarcoplasmic reticulum calcium leak: basis and roles in cardiac dysfunction.
Synchronized SR calcium (Ca) release is critical to normal cardiac myocyte excitation-contraction coupling, and ideally this release shuts off completely between heartbeats. However, other SR Ca release events are referred to collectively as SR Ca leak (which includes Ca sparks and waves as well as smaller events not detectable as Ca sparks). Much, but not all, of the SR Ca leak occurs via ryanodine receptors and can be exacerbated in pathological states such as heart failure. The extent of SR Ca leak is important because it can (a) reduce SR Ca available for release, causing systolic dysfunction; (b) elevate diastolic [Ca]i, contributing to diastolic dysfunction; (c) cause triggered arrhythmias; and (d) be energetically costly because of extra ATP used to repump Ca. This review addresses quantitative aspects and manifestations of SR Ca leak and its measurement, and how leak is modulated by Ca, associated proteins, and posttranslational modifications in health and disease.


Electrophysiological studies on primary cultures of proximal tubule cells.
Primary confluent monolayers were grown from proximal tubule fragments of rabbit kidneys. The fragments were obtained by gradient centrifugation and seeded on an ad hoc dish whose bottom was a permeable and transparent collagen membrane. The culture medium was a mixture of 50% Ham's F-12 and 50% Dulbecco's modified Eagle's medium supplemented with insulin, transferrin, ethanolamine, sodium selenite, and amino acids. The monolayers were studied at 6-14 days after seeding. Transmission electron microscopy revealed cuboidal cells 8.5-10.5 microns high, with a 1.5 to 2.5-microns apical brush border, abundant mitochondria, vacuoles, lysosomes, and irregular basal interdigitating processes. Cyclic AMP synthesis was stimulated by parathyroid hormone and was insensitive to vasopressin and isoproterenol. Electrophysiological studies performed with the same physiological salt solution on both sides revealed a transepithelial voltage of -2.6 +/- 0.6 mV (n = 10) and a basolateral membrane voltage of -51.0 +/- 4.5 mV (n = 13), both referred to the basal solution. The transepithelial electrical resistance was 7 +/- 2 omega X cm2. The apical membrane depolarized on addition of glucose to the apical side and hyperpolarized on removal of glucose. Changes in apical membrane voltage on addition of varying glucose concentrations (at [Na] = 135 mM, 37 degrees C) demonstrate the presence of a glucose transport system with an apparent Km of 3.54 +/- 0.54 and a Vmax of 7.2 +/- 0.4 mV. Thus this preparation exhibits morphological and electrophysiological characteristics of proximal tubule cells; these studies demonstrate the feasibility of the use of intracellular microelectrode techniques to study the transport properties of cultured epithelia.


Reactor design for minimizing product inhibition during enzymatic lignocellulose hydrolysis: II. Quantification of inhibition and suitability of membrane reactors.
Product inhibition of cellulolytic enzymes affects the efficiency of the biocatalytic conversion of lignocellulosic biomass to ethanol and other valuable products. New strategies that focus on reactor designs encompassing product removal, notably glucose removal, during enzymatic cellulose conversion are required for alleviation of glucose product inhibition. Supported by numerous calculations this review assesses the quantitative aspects of glucose product inhibition on enzyme-catalyzed cellulose degradation rates. The significance of glucose product inhibition on dimensioning of different ideal reactor types, i.e. batch, continuous stirred, and plug-flow, is illustrated quantitatively by modeling different extents of cellulose conversion at different reaction conditions. The main operational challenges of membrane reactors for lignocellulose conversion are highlighted. Key membrane reactor features, including system set-up, dilution rate, glucose output profile, and the problem of cellobiose are examined to illustrate the quantitative significance of the glucose product inhibition and the total glucose concentration on the cellulolytic conversion rate. Comprehensive overviews of the available literature data for glucose removal by membranes and for cellulose enzyme stability in membrane reactors are given. The treatise clearly shows that membrane reactors allowing continuous, complete, glucose removal during enzymatic cellulose hydrolysis, can provide for both higher cellulose hydrolysis rates and higher enzyme usage efficiency (kg(product)/kg(enzyme)). Current membrane reactor designs are however not feasible for large scale operations. The report emphasizes that the industrial realization of cellulosic ethanol requires more focus on the operational feasibility within the different hydrolysis reactor designs, notably for membrane reactors, to achieve efficient enzyme-catalyzed cellulose degradation.


Mitochondrial population genomics supports a single pre-Clovis origin with a coastal route for the peopling of the Americas.
It is well accepted that the Americas were the last continents reached by modern humans, most likely through Beringia. However, the precise time and mode of the colonization of the New World remain hotly disputed issues. Native American populations exhibit almost exclusively five mitochondrial DNA (mtDNA) haplogroups (A-D and X). Haplogroups A-D are also frequent in Asia, suggesting a northeastern Asian origin of these lineages. However, the differential pattern of distribution and frequency of haplogroup X led some to suggest that it may represent an independent migration to the Americas. Here we show, by using 86 complete mitochondrial genomes, that all Native American haplogroups, including haplogroup X, were part of a single founding population, thereby refuting multiple-migration models. A detailed demographic history of the mtDNA sequences estimated with a Bayesian coalescent method indicates a complex model for the peopling of the Americas, in which the initial differentiation from Asian populations ended with a moderate bottleneck in Beringia during the last glacial maximum (LGM), around approximately 23,000 to approximately 19,000 years ago. Toward the end of the LGM, a strong population expansion started approximately 18,000 and finished approximately 15,000 years ago. These results support a pre-Clovis occupation of the New World, suggesting a rapid settlement of the continent along a Pacific coastal route.


Feasibility of audit and feedback to reduce postoperative urinary catheter duration.
Indwelling urinary catheter duration is an important risk factor for catheter-associated urinary tract infections (CAUTIs). To audit patient-level postoperative catheter duration and measure the impact of its feedback to nursing staff on postoperative catheter duration and CAUTI rates. Preobservational and postobservational study. Two surgical units at an urban academic medical center. Postoperative patients with indwelling urinary catheters. Audit and feedback of postoperative urinary catheter duration to nurses during an educational session. Patient-level postoperative catheter duration obtained from electronic clinical documentation. Mean catheter duration and proportion of patients with postoperative catheter duration <3 days were determined. Following the intervention, the mean postoperative catheter duration decreased (1.7-1.4 days [P = 0.01] on orthopedic surgery, and 2.6 to 2.2 days [P = 0.01] on general surgery). The proportion of patients with catheter duration <3 days increased significantly on orthopedic surgery (86-92% [P = 0.04]), and nonsignificantly on general surgery (56-63% [P = 0.14]). When adjusted for length of stay differences, the odds of catheter duration <3 days on general surgery increased from 1.38 (P = 0.14) to 1.69 (P = 0.02). The CAUTI rates did not significantly decrease. The rate on orthopedic surgery dropped from 8.9 infections per 1000 device-days to 0 (confidence interval [CI], -1.1 to 18.3); on general surgery the rate was constant at 7 infections per 1000 device-days (CI, -12.1 to 10.8). Audit and feedback of aggregated patient-level urinary catheter duration determined from electronic documentation may prove effective in improving urinary catheter management for surgical patients.


Discrepancies between meta-analyses and subsequent large randomized, controlled trials.
Meta-analyses are now widely used to provide evidence to support clinical strategies. However, large randomized, controlled trials are considered the gold standard in evaluating the efficacy of clinical interventions. We compared the results of large randomized, controlled trials (involving 1000 patients or more) that were published in four journals (the New England Journal of Medicine, the Lancet, the Annals of Internal Medicine, and the Journal of the American Medical Association) with the results of meta-analyses published earlier on the same topics. Regarding the principal and secondary outcomes, we judged whether the findings of the randomized trials agreed with those of the corresponding meta-analyses, and we determined whether the study results were positive (indicating that treatment improved the outcome) or negative (indicating that the outcome with treatment was the same or worse than without it) at the conventional level of statistical significance (P<0.05). We identified 12 large randomized, controlled trials and 19 meta-analyses addressing the same questions. For a total of 40 primary and secondary outcomes, agreement between the meta-analyses and the large clinical trials was only fair (kappa= 0.35; 95 percent confidence interval, 0.06 to 0.64). The positive predictive value of the meta-analyses was 68 percent, and the negative predictive value 67 percent. However, the difference in point estimates between the randomized trials and the meta-analyses was statistically significant for only 5 of the 40 comparisons (12 percent). Furthermore, in each case of disagreement a statistically significant effect of treatment was found by one method, whereas no statistically significant effect was found by the other. The outcomes of the 12 large randomized, controlled trials that we studied were not predicted accurately 35 percent of the time by the meta-analyses published previously on the same topics.


Climate strength: a new direction for climate research.
Climate strength was conceptualized within D. Chan's (1998) discussion of compositional models and the concept of culture strength from the organizational culture literature. Climate strength was operationalized in terms of within-group variability in climate perceptions-the less within-group variability, the stronger the climate. The authors studied climate strength in the context of research linking employee service climate perceptions to customer satisfaction. The hypothesis was tested that climate strength moderates the relationship between employee perceptions of service climate and customer satisfaction experiences. Partial support for the hypothesis was reported in both a concurrent and predictive (3-year) test across 118 branches of a bank. In the predictive study only the interaction of climate and climate strength predicted customer satisfaction. Implications for future research on climate and climate strength are discussed.


Modeling of signaling pathways in chondrocytes based on phosphoproteomic and cytokine release data.
Chondrocyte signaling is widely identified as a key component in cartilage homeostasis. Dysregulations of the signaling processes in chondrocytes often result in degenerative diseases of the tissue. Traditionally, the literature has focused on the study of major players in chondrocyte signaling, but without considering the cross-talks between them. In this paper, we systematically interrogate the signal transduction pathways in chondrocytes, on both the phosphoproteomic and cytokine release levels. The signaling pathways downstream 78 receptors of interest are interrogated. On the phosphoproteomic level, 17 key phosphoproteins are measured upon stimulation with single treatments of 78 ligands. On the cytokine release level, 55 cytokines are measured in the supernatant upon stimulation with the same treatments. Using an Integer Linear Programming (ILP) formulation, the proteomic data is combined with a priori knowledge of proteins' connectivity to construct a mechanistic model, predictive of signal transduction in chondrocytes. We were able to validate previous findings regarding major players of cartilage homeostasis and inflammation (e.g., IL1B, TNF, EGF, TGFA, INS, IGF1 and IL6). Moreover, we studied pro-inflammatory mediators (IL1B and TNF) together with pro-growth signals for investigating their role in chondrocytes hypertrophy and highlighted the role of underreported players such as Inhibin beta A (INHBA), Defensin beta 1 (DEFB1), CXCL1 and Flagellin, and uncovered the way they cross-react in the phosphoproteomic level. The analysis presented herein, leveraged high throughput proteomic data via an ILP formulation to gain new insight into chondrocytes signaling and the pathophysiology of degenerative diseases in articular cartilage.


Copper-mediated cross-coupling of aryl boronic acids and alkyl thiols.
[reaction: see text] The cross-coupling of aryl boronic acids and alkanethiols mediated by copper(II) acetate and pyridine in anhydrous dimethylformamide affords aryl alkyl sulfides in good yield with a wide variety of substituted aryl boronic acids. The method is applicable to the synthesis of aryl sulfides of cysteine.


[Inclusion body myositis].
Sporadic inclusion body myositis (sIBM) is an intractable and progressive skeletal muscle disease of unknown cause that has no curative treatment. Its prevalence varies among countries and ethnic groups. The clinical course is slow and chronic worsening. Diagnosis of sIBM is usually made 5 years after onset. Muscle weakness and atrophy in the quadriceps, wrist flexor, and finger flexors are the typical neurological findings of sIBM. Dysphagia and asymmetric weakness are often found as well. Serum creatine kinase is usually below 2,000 IU/L. Muscle biopsy typically reveals endomysial inflammation, invasion of mononuclear cells into non-necrotic fibers, and rimmed vacuoles, suggesting that inflammation and degeneration are coexist in the pathomechanism. The etiology of sIBM is still unknown; however, genetic factors, aging, lifestyle, and environmental factors may be involved. Recent studies have implicated amyloid beta accumulation, defects of proteolysis, and immune system abnormalities in the pathomechanism of sIBM. sIBM is generally refractory to current therapy, such as steroids or immunosuppressants. Recently, alemtuzumab, which targets T cells, has resulted in improvement in quantitative muscle strength testing. New strategies to induce proteolysis and autophagy, accelerate muscle regeneration, inhibit myostatin, and modulate inflammatory cells are promising. Elucidation of the pathomechanism of sIBM is the key to developing effective therapies.


Prediction model development of late-onset preeclampsia using machine learning-based methods.
Preeclampsia is one of the leading causes of maternal and fetal morbidity and mortality. Due to the lack of effective preventive measures, its prediction is essential to its prompt management. This study aimed to develop models using machine learning to predict late-onset preeclampsia using hospital electronic medical record data. The performance of the machine learning based models and models using conventional statistical methods were also compared. A total of 11,006 pregnant women who received antenatal care at Yonsei University Hospital were included. Maternal data were retrieved from electronic medical records during the early second trimester to 34 weeks. The prediction outcome was late-onset preeclampsia occurrence after 34 weeks' gestation. Pattern recognition and cluster analysis were used to select the parameters included in the prediction models. Logistic regression, decision tree model, naïve Bayes classification, support vector machine, random forest algorithm, and stochastic gradient boosting method were used to construct the prediction models. C-statistics was used to assess the performance of each model. The overall preeclampsia development rate was 4.7% (474 patients). Systolic blood pressure, serum blood urea nitrogen and creatinine levels, platelet counts, serum potassium level, white blood cell count, serum calcium level, and urinary protein were the most influential variables included in the prediction models. C-statistics for the decision tree model, naïve Bayes classification, support vector machine, random forest algorithm, stochastic gradient boosting method, and logistic regression models were 0.857, 0.776, 0.573, 0.894, 0.924, and 0.806, respectively. The stochastic gradient boosting model had the best prediction performance with an accuracy and false positive rate of 0.973 and 0.009, respectively. The combined use of maternal factors and common antenatal laboratory data of the early second trimester through early third trimester could effectively predict late-onset preeclampsia using machine learning algorithms. Future prospective studies are needed to verify the clinical applicability algorithms.


Meconopsis quintuplinervia Regel Improves Cutibacterium acnes-Induced Inflammatory Responses in a Mouse Ear Edema Model and Suppresses Pro-Inflammatory Chemokine Production via the MAPK and NF-κB Pathways in RAW264.7 Cells.
Acne vulgaris (AV) is a common adolescent skin condition which is mainly caused by Cutibacterium acnes overcolonization and subsequent inflammation. Our previous studies demonstrated that ethanol extracts of Meconopsis quintuplinervia Regel (EMQ) possess significant antimicrobial properties. However, their protective effects and potential mechanisms against AV remain unclear. In the present study, the EMQ treatment potential for AV was evaluated in a C. acnes-induced mouse ear edema model, and the EMQ anti-inflammatory mechanism was evaluated in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells. The results showed that EMQ alleviated edema formation and inflammatory cell infiltration in an acne mouse model by suppressing inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor α expression. Moreover, EMQ inhibited the phosphorylation of MAP kinases (MAPKs) such as p38, JNK, and ERK, the phosphorylation and degradation of IκB-α and the nuclear translocation of nuclear factor kappa B (NF-κB) p65 in LPS-induced RAW264.7 cells. These findings suggest the potent anti-inflammatory activity of EMQ is possibly through the regulation of the MAPKs and NF-κB signaling pathways. Inhibition of C. acnes activity combined with a powerful anti-inflammatory effect of EMQ indicated its potential as a novel therapeutic option for AV.


Do insulin and the insulin like growth factors (IGFs) stimulate growth of the exocrine pancreas?
Previous in vivo studies have suggested a long term regulatory role for insulin in the exocrine pancreas. Furthermore, we reported that pancreatic acini have specific receptors for IGF I and II, and using different techniques (acid washing, trypsinisation, electron microscope autoradiography), that CCK8 reduces the internalisation of IGF II. To now directly study the long term role for IGF and insulin in the exocrine pancreas we used AR42J cells, a rat cell line that is derived from a transplantable tumour of the acinar pancreas. Hormone binding studies with 125I-labelled hormones indicated that those cells have insulin receptors, relatively fewer receptors for IGF II but in contrast with normal acini no detectable IGF I receptors. Insulin at concentrations as low as 1 nm stimulated the growth of AR42J cells, as measured by an increase in cell number, DNA and protein content. At 100 nM insulin had maximal effects stimulating the growth by about 50%. IGF I and II had only very weak growth promoting effects probably due to their interaction with the insulin receptor. Additionally insulin increased amylase synthesis over the same concentration range that it stimulated growth. But immunoprecipitation studies revealed that insulin induced a selective increase of amylase synthesis over general protein synthesis. These studies indicate, therefore, that insulin is a growth promoting hormone for AR42J cells and that additionally it seems to specifically regulate amylase synthesis. The role for the IGFs in the exocrine pancreas, however, still remains to be determined.


Age-adjusted and Expanded Lactate Thresholds as Predictors of All-Cause Mortality in the Emergency Department.
While numerous studies have found emergency department (ED) lactate levels to be associated with increased in-hospital mortality, little information is available on the role age plays in this association. This study investigates whether age is a necessary variable to consider when using lactate levels as a marker of prognosis and a guide for management decisions in the ED. This was a retrospective cohort study in an urban, tertiary-care teaching hospital. A total of 13,506 lactate levels were obtained over a 4.5-year period. All adult patients who had a lactate level obtained by the treating provider in the ED were screened for inclusion. The main outcome measure was in-hospital mortality using age-adjusted cohorts and expanded lactate thresholds with secondary outcomes comparing mortality based on the primary clinical impression. Of the 8796 patients in this analysis, there were 474 (5.4%) deaths. Mortality rates increased with both increasing lactate levels and increasing age. For all ages, mortality rates increased from 2.8% in the less than 2.0 millimoles per liter (mmol/L) lactate level, to 5.6% in the 2.0-2.9 mmol/L lactate level, to 8.0% in the 3.0-3.9 mmol/L lactate level, to 13.9% in the 4.0-4.9 mmol/L lactate level, to 13.7% in the 5.0-5.9 mmol/L lactate level, and to 39.1% in the 6.0 mmol/L or greater lactate level (p <0.0001). Survivors, regardless of age, had a mean lactate level <2.0 whereas non-survivors had mean lactate levels of 6.5, 4.5, and 3.7 mmol/L for age cohorts 18-39, 40-64, and ≥ 65 years, respectively. Our findings suggest that although lactate levels can be used as a prognostic tool to risk stratify ED patients, the traditional lactate level thresholds may need to be adjusted to account for varying risk based on age and clinical impressions.


Fitness Consequences of Boldness in Juvenile and Adult Largemouth Bass.
To date, most studies investigating the relationship between personality traits and fitness have focused on a single measure of fitness (such as survival) at a specific life stage. However, many personality traits likely have multiple effects on fitness, potentially operating across different functional contexts and stages of development. Here, we address the fitness consequences of boldness, under seminatural conditions, across life stages and functional contexts in largemouth bass (Micropterus salmoides). Specifically, we report the effect of boldness on (1) juvenile survivorship in an outdoor pond containing natural prey and predators and (2) adult reproductive success in three outdoor ponds across three reproductive seasons (years). Juvenile survival was negatively affected by boldness, with bolder juveniles having a lower probability of survival than shyer juveniles. In contrast, bolder adult male bass had greater reproductive success than their shyer male counterparts. Female reproductive success was not affected by boldness. These findings demonstrate that boldness can affect fitness differently across life stages. Further, boldness was highly consistent across years and significantly heritable, which suggests that boldness has a genetic component. Thus, our results support theory suggesting that fitness trade-offs across life stages may contribute to the maintenance of personality variation within populations.


Hormonal aggressiveness according to the expression of cellular markers in corticotroph adenomas.
The molecular mechanisms underlying tumor growth in Cushing's disease (CD) still remain a challenge. Moreover, clinical manifestations of CD may vary depending on hormonal activity; however, factors involved in the hormonal aggressiveness of adrenocorticotropic hormone (ACTH)-secreting pituitary tumors have not been fully clarified. We investigated the association between the expression of cellular markers regarding pituitary tumor progression and initial or postoperative hormone levels in patients with CD. Tumor tissues from 28 corticotroph adenomas (female 26, male 2, mean age 39.21 ± 10.39 years) were subject to immunohistochemical study using the following antibodies: pituitary tumor-transforming gene 1 (PTTG1), cyclin D1, p16, p27, brahma related-gene 1 (Brg1), and Ki-67. We then analyzed the relationship between each cellular marker expression and hormone levels, including 24 h urinary free cortisol (UFC), plasma ACTH, and serum cortisol. PTTG1 and Ki-67 were expressed in 100% and 50% of patients, respectively. However, the levels did not reflect initial hormonal activity. The cyclin D1-negative group showed higher serum cortisol levels compared to the cyclin D1-positive group (p = 0.01). The 24 h UFC levels were significantly higher in the p27-negative group than in the p27-positive group (p = 0.04), whereas the Brg1-positive group revealed higher serum cortisol levels than in the Brg1-negative group (p = 0.02). Although PTTG1 and Ki-67 play an essential role in developing ACTH-secreting tumors, cyclin D1, p27, and Brg1 may be better biomarkers to determine hormonal aggressiveness of the tumor. Further research is needed to understand the influence of cellular markers on hormonal activity in CD.


Hierarchical cluster analysis of immunophenotype classify AML patients with NPM1 gene mutation into two groups with distinct prognosis.
The prognostic implication of immunophenotyping in acute myeloid leukemia (AML) patients with NPM1 mutation remains unclear. Ninety-four of 543 AML patients diagnosed with NPM1 mutation between 1987 and 2007 were studied. The expression of surface antigens on leukemic cells was evaluated with respect to clinical manifestations and outcomes. In order to validate the prognostic effect of the immunophenotypic cluster, another 36 patients with NPM1 mutation diagnosed between 2008 and 2010 were analyzed. Ninety-four patients with NPM1 mutations and complete immunophenotyping data were enrolled for a hierarchical cluster analysis and the result was correlated with clinico-laboratory characteristics. Clustering analysis divided the patients with NPM1 mutations into the following two groups: group I, CD34(-)/CD7(-), but with variable expression of HLA-DR; and group II, HLA DR(+)/CD34(+)/CD7(+). With a median follow-up of 53 months, the group II patients had a significantly shorter relapse-free survival (RFS, median: 3 vs. 23 months, p = 0.006) and overall survival (OS, median: 11 vs. 40 months, p = 0.02) than group I patients. Multivariate analysis of variables, including clinico-laboratory data and other gene mutations revealed that the immunophenotypic cluster is an independent prognostic factor (RFS, p = 0.002; OS, p = 0.024). In order to confirm the prognostic effect of the immunophenotypic cluster, another 36 patients with NPM1 mutation diagnosed between 2008 and 2010 were validated. Hierarchical cluster analysis also showed two distinct clusters, group I patient showed significant better RFS (p = 0.021), and OS (p = 0.055). In total, we stratified 130 NPM1-mutant patients, by FLT3-ITD mutation and immunophenotypic cluster into distinct prognostic groups (RFS, p < 0.001 and OS, p = 0.017). Among NPM1-mutated AML, the antigen expression pattern of HLADR(+) CD34(+) CD7(+) is associated with a poor prognosis, independent to the FLT3-ITD mutation.


Arterial blood pressure and cardiovascular responses to yoga practice.
Yoga is qualitatively different from any other mode of physical activity in that it consists of a unique combination of isometric muscular contractions, stretching exercises, relaxation techniques, and breathing exercises. In particular, yoga postures consist of systemic isometric contractions that are known to elicit marked increases in mean blood pressure that are not observed during dynamic exercise. Stretching can also induce increases in blood pressure and sympathetic nerve activity in the muscles. Currently, not much is known about changes in blood pressure and other cardiovascular responses to yoga practice. The study intended to determine the acute effects of one session of hatha yoga practice on blood pressure and other cardiovascular responses. To gain insight into the long-term effects of yoga practice, both novice (n = 19) and advanced (n = 18) yoga practitioners were studied. The two groups were matched for age, gender, BMI, and blood pressure. The setting was a research laboratory at a university. Thirty-six apparently healthy, nonobese, sedentary, or recreationally active individuals from the community participated in the study. Intervention The intervention comprised one session of yoga practice, in which participants followed a custom made instructional video providing a yoga routine that consisted of a series of 23 hatha-based yoga postures. Prior to arriving at the laboratory, each participant completed a research health questionnaire, a training-status questionnaire, and a yoga-experience questionnaire. Prior to the yoga practice, each participant's height, body fat percentage, trunk or lumbar flexibility, and arterial stiffness as assessed by carotid femoral pulse wave velocity (cfPWV) were measured. For each posture during the yoga practice, the study continuously measured systolic, mean, and diastolic blood pressures, heart rate, stroke volume, and cardiac output. Systolic, mean, and diastolic blood pressures increased significantly during the yoga practice. The magnitude of these increases in blood pressure was greatest with standing postures. Heart rate and cardiac output increased significantly during yoga practice, especially with standing postures. Overall, no differences existed in cardiovascular responses between the novice and advanced practitioners throughout the yoga testing session; cfPWV velocity was significantly and inversely associated with lumbar flexion but not with sit-and-reach test scores. The research team concluded that a variety of hatha yoga postures, especially standing postures, evoked significant increases in blood pressure. The elevation in blood pressure due to yoga practice was associated with increases in cardiac output and heart rate, which are responses similar to those observed in isometric exercise. The lack of obvious differences in blood pressure and other cardiovascular responses between novice and advanced yoga practitioners suggests that long-term yoga practice does not attenuate acute yoga responses.


Short-term adaptations of the dynamic disparity vergence and phoria systems.
The ability to adapt is critical to survival and varies between individuals. Adaptation of one motor system may be related to the ability to adapt another. This study sought to determine whether phoria adaptation was correlated with the ability to modify the dynamics of disparity vergence. Eye movements from ten subjects were recorded during dynamic disparity vergence modification and phoria adaptation experiments. Two different convergent stimuli were presented during the dynamic vergence modification experiment: a test stimulus (4° step) and a conditioning stimulus (4° double step). Dynamic disparity vergence responses were quantified by measuring the peak velocity (°/s). Phoria adaptation experiments measured the changes in phoria over a 5-min period of sustained fixation. The maximum velocity of phoria adaptation was determined from an exponential fit of the phoria data points. Phoria and dynamic disparity vergence peak velocity were both significantly modified (P < 0.001). The maximum velocity of phoria adaptation was significantly correlated with the changes in convergence peak velocity (r > 0.89; P < 0.001). There was a strong correlation between the ability to adaptively adjust two different oculomotor parameters: a tonic and dynamic component. Future studies should investigate additional interactions between these parameters, and the ability to adaptively change other oculomotor systems such as the saccadic or smooth pursuit system. Understanding the ability to modify phoria, dynamic disparity vergence, and other oculomotor parameters can yield insights into the plasticity of short-term adaptation mechanisms.


A new microdialysis probe for continuous lactate measurement during fetal monitoring: Proof of concept in an animal model.
Cardiotocography (CTG) is currently the most commonly used method for intrapartum fetal monitoring during labor. However, a high false-positive rate of fetal acidosis indicated by CTG leads to an increase in obstetric interventions. We developed a microdialysis probe that is integrated into a fetal scalp electrode allowing continuous measurement of lactate subcutaneously, thus giving instant information about the oxygenation status of the fetus. Our aim was to establish proof of concept in an animal model using a microdialysis probe to monitor lactate subcutaneously. We performed an in vivo study in adult male wild-type Wistar rats. We modified electrodes used for CTG monitoring in human fetuses to incorporate a microdialysis membrane. Optimum flow rates for microdialysis were determined in vitro. For the in vivo experiment, a microdialysis probe was inserted into the skin on the back of the animal. De-oxygenation and acidosis were induced by lowering the inspiratory oxygen pressure. Oxygenation and heart rate were monitored. A jugular vein cannula was inserted to draw blood samples for analysis of lactate, pH, pco2 , and saturation. Lactate levels in dialysate were compared with plasma lactate levels. Baseline blood lactate levels were around 1 mmol/L. Upon de-oxygenation, oxygen saturation fell to below 40% for 1 h and blood lactate levels increased 2.5-fold. Correlation of dialysate lactate levels with plasma lactate levels was 0.89 resulting in an R2 of .78 in the corresponding linear regression. In this animal model, lactate levels in subcutaneous fluid collected by microdialysis closely reflected blood lactate levels upon transient de-oxygenation, indicating that our device is suitable for subcutaneous measurement of lactate. Microdialysis probe technology allows the measurement of multiple compounds in the dialysate, such as glucose, albumin, or inflammatory mediators, so this technique may offer the unique possibility to shed light on fetal physiology during the intrapartum period.


Anti-skin-aging effect of epigallocatechin gallate by regulating epidermal growth factor receptor pathway on aging mouse model induced by d-Galactose.
Epigallocatechin gallate(EGCG) is a monomer separated from tea catechins, as an well-known antioxidant, which helps fight wrinkles and rejuvenate skin cells. In this study, we investigated the anti-aging effect of EGCG, and to clarify underlying mechanism of skin aging in a d-galactose-induced aging mouse model. Forty-five male mice were divided into 5 groups and treated with different dose of EGCG, Vitamin C (VitC) to mice as a positive control. All groups except vehicle were established aging model induced by d-galactose (200mg/kg/day) that was subcutaneously injected to mice for 8 weeks. Two weeks after injection of d-galactose, EGCG and Vit C groups were simultaneously administered once a day by subcutaneously inject after 5h for injecting d-galactose. The results show that EGCG can be absorbed by the skin. Overall, the conditions of the skin of EGCG-treatment groups were improved, the whole structure of skin were better than control groups, and the levels of oxidative stress and the expression of relate with EGFR proteins were significantly higher than control group after EGCG treatment. All these findings suggest that EGCG can resist skin senility effectively. And the EGFR with relate of downstream proteins are implicated in the skin aging.


Potential mechanism of estrogen-mediated decrease in bone formation: estrogen increases production of inhibitory insulin-like growth factor-binding protein-4.
Using a recently developed human osteoblastic cell line (hFOB/ER9) with high levels (approximately 4,000 per nucleus) of estrogen receptors and the characteristic phenotype of the mature osteoblast, we tested the hypothesis that estrogen decreases bone formation by inhibiting the action of the insulin-like growth factor (IGF) paracrine/autocrine system. IGF-II, the predominant IGF produced by osteoblastic cells, was measurable in hFOB/ER9-conditioned medium (approximately 10 ng/mL) and its level did not change significantly after treatment with 17 beta-estradiol (E2) or anti-estrogens. Treatment with E2 at 0.1-100 nM decreased [3H]thymidine uptake to 53% of control (p < 0.001) in a dose-dependent fashion. The predominant IGF-binding proteins (IGFBPs) produced by hFOB/ER9 and by normal trabecular osteoblasts are IGFBP-3 and IGFBP-4, of which IGFBP-4 is consistently inhibitory of IGF action. Treatment with E2 at 0.01-10 nM for 48 h increased IGFBP-4 mRNA to 346% +/- 90% (mean +/- SE) of control (p < 0.05) and IGFBP-4 protein to 278% +/- 75% of control (p < 0.01) in a dose-dependent fashion but did not alter IGFBP-3 mRNA or protein. E2 treatment also attenuated IGF-dependent, IGFBP-4 specific proteolysis to approximately 50% of control. ICI 182,780, a pure anti-estrogen, completely blocked E2-mediated decreases in cell proliferation and increases in levels of IGFBP-4 mRNA and protein. Treatment of the hFOB/ER9 cells with recombinant human IGFBP-4 (200 ng/mL) decreased cell proliferation to 55% of control (p < 0.01). Thus, E2 acts on osteoblastic cells to increase availability of inhibitory IGFBP-4, by both increasing its production and decreasing its degradation, which may oppose the mitogenic effect of the IGFs on osteoblastic cells. This action may mediate, at least in part, the decreases in bone formation that are observed after estrogen treatment in vivo.


White-matter development is different in bilingual and monolingual children: a longitudinal DTI study.
Although numerous people grow up speaking more than one language, the impact of bilingualism on brain developing neuroanatomy is still poorly understood. This study aimed to determine whether the changes in the mean fractional-anisotropy (MFA) of language pathways are different between bilingual and monolingual children. Simultaneous-bilinguals, sequential-bilinguals and monolingual, male and female 10-13 years old children participated in this longitudinal study over a period of two years. We used diffusion tensor tractography to obtain mean fractional-anisotropy values of four language related pathways and one control bundle: 1-left-inferior-occipitofrontal fasciculus/lIFOF, 2-left-arcuate fasciculus/lAF/lSLF, 3-bundle arising from the anterior part of corpus-callosum and projecting to orbital lobe/AC-OL, 4-fibres emerging from anterior-midbody of corpus-callosum (CC) to motor cortices/AMB-PMC, 5- right-inferior-occipitofrontal fasciculus rIFOF as the control pathway unrelated to language. These values and their rate of change were compared between 3 groups. FA-values did not change significantly over two years for lAF/lSLF and AC-OL. Sequential-bilinguals had the highest degree of change in the MFA value of lIFOF, and AMB-PMC did not present significant group differences. The comparison of MFA of lIFOF yielded a significantly higher FA-value in simultaneous bilinguals compared to monolinguals. These findings acknowledge the existing difference of the development of the semantic processing specific pathway between children with different semantic processing procedure. These also support the hypothesis that age of second language acquisition affects the maturation and myelination of some language specific white-matter pathways.


Plantation medicine in Hawaii 1840 to 1964: a patient's perspective.
The first contract laborers arrived from China in 1852, but little attention was paid to their medical needs. In 1886 a physician from Japan arrived to minister to the health and medical needs of the Japanese immigrants. After 1900 most physicians caring for immigrant plantation workers were Western trained from the Mainland. Many advances in medicine were started on plantations such as the second x-ray machine in Hawaii. The 1950s brought rapid changes in plantation medicine with the closing of plantation facilities on Oahu. The author describes her personal experience as a patient in the plantation medical system.


Postoperative Outcomes of Branchial Cleft Cyst Excision in Children and Adults: An NSQIP Analysis.
Identify risk factors and perioperative morbidity for patients undergoing branchial cleft cyst (BCC) excision. Cross-sectional analysis. American College of Surgeons National Surgical Quality Improvement Program adult and pediatric databases (NSQIP and NSQIP-P). Patients who underwent BCC excision (Current Procedural Terminology 42810, 42815) were queried via NSQIP (2005-2016) and NSQIP-P (2012-2016). Outcomes analyzed include patient demographics, medical comorbidities, admission type, operative characteristics, length of hospital stay, postoperative complications, and readmission. A total of 1775 children and 677 adults were identified. Mean age at time of surgery was 4.6 years for children and 38.6 years for adults. Outpatient procedures were performed in 87.1% of adults and 94.0% of children (P < .001). Postoperative complications were uncommon, occurring in <1% of adults and 3.9% of children (P < .001). Similarly, readmission occurred in 1.2% of adults and 1.1% of children. In adults, smoking status was shown to have a significant effect on postoperative complications (odds ratio, 6.25; P = .037). Age group did not have an effect on the complication rate in the pediatric population. Pediatric otolaryngologists had higher rates of postoperative complications (P = .001), prolonged operative times (P < .001), and fewer outpatient procedures (P < .001). Conversely, in adults, otolaryngologists had fewer postoperative complications. Postoperative complications following BCC excision are relatively uncommon, demonstrating procedural safety when performed at any age.


Heat stress decreases testicular germ cell proliferation and increases apoptosis in short term: an immunohistochemical and ultrastructural study.
Scrotal hyperthermia has been known as a cause of male infertility but the exact mechanism leading to impaired spermatogenesis is unknown. This work was aimed to investigate the role of scrotal hyperthermia on cell proliferation and apoptosis in testes. The rats were randomly allotted into one of the four experimental groups: A (control), B (1 day after scrotal hyperthermia), C (14 days after scrotal hyperthermia), and D (35 days after scrotal hyperthermia); each group comprised 7 animals. Scrotal hyperthermia was carried out in a thermostatically controlled water bath at 43°C for 30 min once daily for 6 consecutive days. Control rats were treated in the same way, except the testes were immersed in a water bath maintained at 22°C. Hyperthermia-exposed rats were killed under 50 mg/kg ketamine anaesthesia and tissue samples were obtained for biochemical and histopathological investigations. Hyperthermia treatment significantly decreased the testicular antioxidant system, including decreases in the glutathione level, superoxide dismutase, and glutathione peroxidase activities. Moreover, exposure to hyperthermia resulted in lipid peroxidation increase in testes. Our data indicate a significant reduction in the expression of proliferating cell nuclear antigen and an enhancement in the activity of terminal deoxynucleotidyl transferase dUTP nick end labelling after scrotal hyperthermia. In scrotal hyperthermia, the mitochondrial degeneration, dilatation of smooth endoplasmic reticulum, and enlarged intercellular spaces were observed in both Sertoli and spermatid cells. Scrotal hyperthermia is one of the major factors that impair spermatogenesis in testis. This heat stress is shown to be closely associated with oxidative stress, followed by apoptosis of germ cells.


Expression of growth differentiation factor-5 and bone morphogenic protein-7 in intraocular osseous metaplasia.
Intraocular bone is seen in a wide spectrum of ocular disorders. The pathogenetic mechanisms of bone formation in the eye are unclear. Growth differentiation factor-5 (GDF-5), bone morphogenic protein-7 (BMP-7), and transforming growth factor beta-1 (TGF beta1) are multifunctional cytokines that have important roles in bone formation. Immunohistochemistry was used to localise GDF-5, BMP-7, and TGF beta1 in the human eye to determine their role in intraocular bone formation. Paraffin embedded sections from human eyes included fetal eyes (n = 5), normal adult eyes (n = 4), eyes with osseous metaplasia (n = 8), and eyes with focal fibrous metaplasia of the retinal pigment epithelium (RPE) without osseous metaplasia (n = 2). Immunohistochemistry was performed using indirect immunofluorescence with antibodies to GDF-5, BMP-7, and TGF beta1. The staining intensity was evaluated semiquantitatively in the RPE, retina, ciliary epithelium, and cornea; and analysed statistically. When compared with normal adult eyes, which showed no RPE immunoreactivity, the RPE metaplasia surrounding areas of osseous metaplasia showed mild GDF-5 and moderate BMP-7 (p = 0.004) intracytoplasmic immunoreactivity. In contrast, trace GDF-5 and mild BMP-7 staining was seen in zones of RPE fibrous metaplasia in areas not associated with osseous metaplasia. Mild intracytoplasmic TGF beta1 expression was seen in the RPE metaplasia surrounding the bone when compared with adult eyes. Both fetal and adult eyes showed trace to mild GDF-5 and BMP-7 labelling of the non-pigmented ciliary epithelium which was increased in the eyes with osseous metaplasia. In eyes with osseous metaplasia, a significant decrease in GDF-5 and BMP-7 labelling was noted in fetal keratocytes (p = 0.0159 for both antibodies) when compared to adult eyes. Also, a significant decrease in BMP-7 labelling was seen in keratocytes in eyes with osseous metaplasia (p = 0.0162). The increase in GDF-5, BMP-7, and TGF beta1 immunoreactivity in zones of RPE metaplasia in eyes with osseous metaplasia suggests that these proteins have an important role in intraocular ectopic bone formation.


Late presenters to HIV care and treatment, identification of associated risk factors in HIV-1 infected Indian population.
Timely access to antiretroviral therapy is a key to controlling HIV infection. Late diagnosis and presentation to care diminish the benefits of antiretrovirals and increase risk of transmission. We aimed to identify late presenters in patients sent for first CD4 T cell count after HIV diagnosis, for therapy initiation evaluation. Further we aimed at identifying patient factors associated with higher risk of late presentation. Retrospective data collection and analysis was done for 3680 subjects visiting the laboratory for CD4 T cell counts between 2001 and 2007. We segregated the patients on basis of their CD4 T cell counts after first HIV diagnosis. Factors associated with risk of late presentation to CD4 T cell counts after HIV diagnosis were identified using univariate analysis, and the strength of association of individual factor was assessed by calculation of odds ratios. Of 3680 subjects, 2936 (83.37%) were defined as late presenters. Late testing varied among age groups, transmission categories, and gender. Males were twice as likely to present late as compared to females. We found significant positive association of heterosexual transmission route (p < 0.001), and older age groups of 45 years and above (p = 0.0004) to late presentation. Female sex, children below 14 years of age and sexual contact with HIV positive spouse were associated with significantly lower risks to presenting late. Intravenous drug users were also associated with lower risks of late presentation, in comparison to heterosexual transmission route. The study identifies HIV infected population groups at a higher risk of late presentation to care and treatment. The risk factors identified to be associated with late presentation should be utilised in formulating targeted public health interventions in order to improve early HIV diagnosis.


Molecular Regulation of Plant Responses to Environmental Temperatures.
Temperature is a key factor governing the growth and development, distribution, and seasonal behavior of plants. The entire plant life cycle is affected by environmental temperatures. Plants grow rapidly and exhibit specific changes in morphology under mild average temperature conditions, a response termed thermomorphogenesis. When exposed to chilling or moist chilling low temperatures, flowering or seed germination is accelerated in some plant species; these processes are known as vernalization and cold stratification, respectively. Interestingly, once many temperate plants are exposed to chilling temperatures for some time, they can acquire the ability to resist freezing stress, a process termed cold acclimation. In the face of global climate change, heat stress has emerged as a frequent challenge, which adversely affects plant growth and development. In this review, we summarize and discuss recent progress in dissecting the molecular mechanisms regulating plant thermomorphogenesis, vernalization, and responses to extreme temperatures. We also discuss the remaining issues that are crucial for understanding the interactions between plants and temperature.


[Hysterosalpingography: synechiae? Gartner's ducts?].
A linear shadow parallel to the uterus longitudinal axis, was observed in a hysterosalpingography (HSG) and has set up a controversy between synechiae and Gartner's ducts, without reaching a definitive conclusion. A case like this does not seem to have been published previously.


Phorbol esters inhibit gluconeogenesis in canine renal proximal tubular segments.
Gluconeogenesis is a major metabolic function of the renal proximal tubular cell. To characterize the regulation of this process in proximal tubule, glucose production from gluconeogenic precursors was measured in proximal tubular segments prepared from dog kidney. Production of glucose was a linear function of time for up to 120 min of incubation at 37 degrees C under a variety of conditions. Lowering the pH of incubation media from 7.5 to 7.0 increased glucose synthesis. Production of glucose was inhibited by 3-mercaptopicolinate. Incubation of proximal tubular segments with insulin diminished synthesis of glucose. Incubation of segments with tumor-promoting phorbol esters, 12-O-tetradecanoylphorbol-13-acetate or phorbol 12,13-dibutyrate, resulted in decreased production of glucose. This effect was not observed following incubation with the inactive phorbol ester 4 alpha-phorbol. Changes in glucose synthesis could not be attributed to alterations in cell viability or in rates of glucose oxidation induced by experimental maneuvers. Our findings confirm the usefulness of proximal tubular segments for characterization of metabolic processes in this portion of the nephron. The experimental results are consistent with a role for protein kinase C in the control of gluconeogenesis in proximal tubule.


[Kinetics and immunocytochemical staining of 99mTc-labeled monoclonal anti-CEA antibodies against granulocytes following intravenous administration].
Until now the clinical identification of the affinity of monoclonal 99mTc-anti-CEA antibodies (MAK BW 250/183) on granulocytes was made with tumor cells carrying the same epitope on NCA-95 and human granulocytes in vitro. As this antibody only binds human granulocytes, animal experiments are impossible. 3 patients had their blood withdrawn within 6 h after injection, another patient had his left hip-joint biopsied after 24 h, the samples undergoing subsequent immunocytochemical dyeing. Dyeing of granulocytes all over the smears was evident whereas lymphocytes, monocytes and erythrocytes did not show any reaction. After 6 h there seemed to be a large difference between a relatively high quantity of 86% unlabelled 99mTc-MAb and 75% of immunocytochemically stainable granulocytes in the blood through an excess of binding epitopes. Six h after injection 27% of the activity were, on average, detectable in whole blood. At this time the activity in blood was reduced to an extent that scintigraphic imaging was feasible.


In vivo and in silico pharmacokinetics and biodistribution of a melanocortin receptor 1 targeted agent in preclinical models of melanoma.
The melanocortin 1 receptor (MC1R) is overexpressed in most melanoma metastases, making it a promising target for imaging of melanomas. In this study, the expression of MC1R in a large fraction of patients with melanoma was confirmed using mRNA and tissue microarray. Here, we have characterized the in vivo tumor and tissue distribution and pharmacokinetics (PK) of uptake and clearance of a MC1R specific peptidomimetic ligand conjugated to a near-infrared fluorescent dye. We propose an interdisciplinary framework to bridge the different time and space scales of ligand-tumor-host interactions: intravital fluorescence microscopy to quantify probe internalization at the cellular level, a xenograft tumor model for whole body pharmacokinetics, and a computational pharmacokinetic model for integration and interpretation of experimental data. Administration of the probe into mice bearing tumors with high and low MC1R expression demonstrated normalized image intensities that correlated with expression levels (p < 0.05). The biodistribution study showed high kidney uptake as early as 30 min postinjection. The PK computational model predicted the presence of receptors in the kidneys with a lower affinity, but at higher numbers than in the tumors. As the mouse kidney is known to express the MC5R, this hypothesis was confirmed by both coinjection of a ligand with higher MC5R affinity compared to MC1R and by injection of lower probe concentrations (e.g., 1 nmol/kg), both leading to decreased kidney accumulation of the MC1R ligand. In addition, through this interdisciplinary approach we could predict the rates of ligand accumulation and clearance into and from organs and tumors, and the amount of injected ligand required to have maximum specific retention in tumors. These predictions have potential to aid in the translation of a targeted agent from lab to the clinic. In conclusion, the characterized MC1R-specific probe has excellent potential for in vivo detection of melanoma metastases. The process of cell-surface marker validation, targeted imaging probe development, and in vitro, in vivo, and in silico characterization described in this study can be generally applied to preclinical development of targeted agents.


Effect of germanium doping on pyroelectric and piezoelectric properties of Sn(2)P(2)S(6) single crystal.
The results of the search for dopants to optimize the ferroelectric phase transition temperature and pyroelectric and piezoelectric properties of Sn(2)P(2)S(6) crystals are reported. Among all dopants only germanium causes a pronounced shift of the phase transition toward higher temperatures. The highest Curie temperature achieved by Ge doping is as high as 88 degrees C. The major advantage of the Ge-doped crystals as compared to pure Sn(2)P(2)S(6) is that the hydrostatic piezoelectric and pyroelectric response is kept high in a wider temperature range with much lower temperature dependence. The effects of the doping on pyroelectric and piezoelectric properties, characterizing the sensitivity of the material to thermal and hydroacoustic excitation, are discussed.


Structure-Guided Strategy for the Development of Potent Bivalent ERK Inhibitors.
ERK is the effector kinase of the RAS-RAF-MEK-ERK signaling cascade, which promotes cell transformation and malignancy in many cancers and is thus a major drug target in oncology. Kinase inhibitors targeting RAF or MEK are already used for the treatment of certain cancers, such as melanoma. Although the initial response to these drugs can be dramatic, development of drug resistance is a major challenge, even with combination therapies targeting both RAF and MEK. Importantly, most resistance mechanisms still rely on activation of the downstream effector kinase ERK, making it a promising target for drug development efforts. Here, we report the design and structural/functional characterization of a set of bivalent ERK inhibitors that combine a small molecule inhibitor that binds to the ATP-binding pocket with a peptide that selectively binds to an ERK protein interaction surface, the D-site recruitment site (DRS). Our studies show that the lead bivalent inhibitor, SBP3, has markedly improved potency compared to the small molecule inhibitor alone. Unexpectedly, we found that SBP3 also binds to several ERK-related kinases that contain a DRS, highlighting the importance of experimentally verifying the predicted specificity of bivalent inhibitors. However, SBP3 does not target any other kinases belonging to the same CMGC branch of the kinome. Additionally, our modular click chemistry inhibitor design facilitates the generation of different combinations of small molecule inhibitors with ERK-targeting peptides.


Expression of lumican and fibromodulin following interleukin-1 beta stimulation of disc cells of the human temporomandibular joint.
Small leucine-rich repeat proteoglycans (SLRP) are present in the extracellular matrix of the temporomandibular joint (TMJ) disc. Lumican and fibromodulin, classified as class 2 SLRPs, play important roles in TMJ assembly, proliferation and inflammation. Degenerative change in the TMJ disc gives rise to the process of internal derangement (ID). In this study, we immunohistochemically examined the expression of lumican and fibromodulin in nine human TMJ specimens and examined the gene expression of both proteoglycans in cultured human TMJ disc cells under interleukin-1 beta (IL-1 β)-stimulated conditions. An articular disc cell line was established by collagenase treatment of a TMJ disc. The subcultured cells were then incubated for 1, 3, 6, 12, 24 or 48 h under both normal and IL-1 β (1 ng/mL) conditions. The gene expression of lumican and fibromodulin was examined using the reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR. We demonstrated that the expression of lumican significantly differs from that of fibromodulin in the deformed disc and that IL-1 β induces a significant increase in lumican mRNA, but not in fibromodulin mRNA, after 24∼48 h culture compared to cells cultured in the absence of IL-1 β (P<0.05). These results indicate that lumican and fibromodulin display different behaviors and that lumican may promote regeneration of the TMJ after degeneration and deformation induced by IL-1 β.


Exosomes in the Treatment of Pancreatic Cancer: A Moonshot to PDAC Treatment?
Pancreatic Ductal Adenocarcinoma (PDAC) constitutes a leading cause of cancer death globally. Its mortality remains unaltered despite the considerable scientific progress made in the fields of diagnostics and treatment. Exosomes comprise of small extracellular vesicles secreted by nearly all cells; their cargo contains a vast array of biomolecules, such as proteins and microRNAs. It is currently established that their role as messengers is central to a plethora of both physiologic and pathologic processes. Accumulating data have shed light on their contributions to carcinogenesis, metastasis, and immunological response. Meanwhile, the advancement of personalized targeted therapies into everyday clinical practice necessitates the development of cost-efficient treatment approaches. The role of exosomes is currently being extensively investigated towards this direction. This review aims to summarize the current pre-clinical and clinical evidence regarding the effects of exosomal applications in the timely diagnosis, prognosis, and therapeutic management of pancreatic cancer.


Impact of Early and Late Visual Deprivation on the Structure of the Corpus Callosum: A Study Combining Thickness Profile with Surface Tensor-Based Morphometry.
Blindness represents a unique model to study how visual experience may shape the development of brain organization. Exploring how the structure of the corpus callosum (CC) reorganizes ensuing visual deprivation is of particular interest due to its important functional implication in vision (e.g., via the splenium of the CC). Moreover, comparing early versus late visually deprived individuals has the potential to unravel the existence of a sensitive period for reshaping the CC structure. Here, we develop a novel framework to capture a complete set of shape differences in the CC between congenitally blind (CB), late blind (LB) and sighted control (SC) groups. The CCs were manually segmented from T1-weighted brain MRI and modeled by 3D tetrahedral meshes. We statistically compared the combination of local area and thickness at each point between subject groups. Differences in area are found using surface tensor-based morphometry; thickness is estimated by tracing the streamlines in the volumetric harmonic field. Group differences were assessed on this combined measure using Hotelling's T(2) test. Interestingly, we observed that the total callosal volume did not differ between the groups. However, our fine-grained analysis reveals significant differences mostly localized around the splenium areas between both blind groups and the sighted group (general effects of blindness) and, importantly, specific dissimilarities between the LB and CB groups, illustrating the existence of a sensitive period for reorganization. The new multivariate statistics also gave better effect sizes for detecting morphometric differences, relative to other statistics. They may boost statistical power for CC morphometric analyses.


In situ proliferative epithelial lesions of the breast.
Considerable progress has been made over the last several years toward understanding the significance of noninvasive epithelial changes in the breast. This progress, in essence, is the result of categorizing in situ alterations according to their clinical significance, or more specifically, according to the risk that such lesions carry to the patient for development of a subsequent invasive carcinoma. This article summarizes the histopathological criteria that are currently used to define morphologic categories and, perhaps more importantly, it reviews the clinical implications of the diagnoses that result from this categorization.


Iron supplementation for the control of iron deficiency in populations at risk.
Iron supplementation, mostly with a therapeutic orientation, has been a key strategy for the short-term control of iron deficiency and ferropenic anemia. It has been used almost exclusively in antenatal clinics, but in spite of its confirmed efficacy in supervised trials, it has proven ineffective in practice in most developing countries. Poor effectiveness has been attributed to various factors including insufficient dose and time of supplementation and poor adherence. These problems have led to the administration of high iron doses, which have proven equally ineffective in practice. This paper introduces four concepts: (1) that iron supplementation targeted to pregnant women should cover the full reproductive cycle, from prepregnancy to at least the end of lactation instead of only the pregnant women; (2) that entering pregnancy with iron deficiency contributes to the failure of antenatal iron supplementation and that prepregnancy iron reserves increase the effectiveness of antenatal supplementation; (3) that medium- to long-term weekly ingestion of proper iron-folate supplements, with a preventive aim and directed to all risk groups, should be community based rather than health service based but supervised by the latter (in this sense, preventive supplementation is equal to targeted iron fortification); and (4) that preventive supplementation, based on weekly dosing, has proven efficacious. Problem-oriented research to evaluate the sustainability and medium- to long-term efficacy of these concepts is called for. The bases for the concepts and suggestions are summarized in this paper.


Stiffness and ankylosis in primary total knee arthroplasty.
To do a successful total knee arthroplasty (TKA), adequate exposure of the tibial plateau and distal femur is required. Difficulty with exposure often is encountered in the patient with a stiff or ankylosed knee. This can lead to complications including component malpositioning and extensor mechanism problems, such as patellar tendon rupture. In these cases where the knee has limited motion preoperatively, knowledge of advanced techniques for soft tissue treatment, and for optimizing exposure are required. Numerous such techniques have been described, including the quadriceps snip, V-Y quadriceps turndown, tibial tubercle osteotomy, and femoral peel. A systematic review of these techniques, including indications and results, based published reports and our experiences, is presented.


[Nutritional repercussions and management of chronic pancreatitis].
The pancreas is a retroperitoneal organ that releases water, bicarbonate and digestive enzymes by the main pancreatic duct (MPD) into the duodenum. Chronic pancreatitis (CP) is typically caused, in adults, by chronic alcohol abuse and, less frequently hypertriglyceridemia, primary hyperparathyroidism or cystic fibrosis. Exocrine dysfunction results in malabsorption of fat and subsequent steatorrhea. Damage to pancreatic endocrine function is a late finding in CP and results in hyperglycaemia or overt diabetes mellitus. Care of patients with CP principally involves management of pain. A significant change in the pain pattern or the sudden onset of persistent symptoms suggests the need to rule out other potential etiologies, including peptic ulcer disease, biliary obstruction, pseudocysts, pancreatic carcinoma, and pancreatic duct stricture or stones, then is important to establish a secure diagnosis. Management of pain should then proceed in a judicious stepwise approach avoiding opioids dependence. Patients should be advised to stop alcohol intake. Fat malabsorption and other complications may also arise. Management of steatorrhea should begin with small meals and restriction in fat intake. Pancreatic enzyme supplements can relieve symptoms and reduce malabsorption in patients who do not respond to dietary restriction. Enzymes at high doses should be used with meals. Treatment with acid suppression to reduce inactivation of the enzymes from gastric acid are recommended. Supplementation with medium chain triglycerides and fat soluble vitamin replacement may be required. Management of other complications (such as pseudocysts, bile duct or duodenal obstruction, pancreatic ascites, splenic vein thrombosis and pseudoaneurysms) often requires aggressive approach with the patient kept on total parenteral nutrition to minimize pancreatic stimulation.


Multiscale Functional Imaging of Interfaces through Atomic Force Microscopy Using Harmonic Mixing.
The spatial resolution of atomic force microscopy (AFM) needed to resolve material interfaces is limited by the tip-sample separation ( d) dependence of the force used to record an image. Here, we present a new multiscale functional imaging technique that allows for in situ tunable spatial resolution, which can be applied to a wide range of inhomogeneous materials, devices, and interfaces. Our approach uses a multifrequency method to generate a signal whose d-dependence is controlled by mixing harmonics of the cantilever's oscillation with a modulated force. The spatial resolution of the resulting image is determined by the signal's d-dependence. Our measurements using harmonic mixing (HM) show that we can change the d-dependence of a force signal to improve spatial resolution by up to a factor of two compared to conventional methods. We demonstrate the technique with both Kelvin probe force microscopy (KPFM) and bimodal AFM to show its generality. Bimodal AFM with harmonic mixing actuation separates conservative from dissipative forces and is used to identify the regions of adhesive residue on exfoliated graphene. Our electrostatic measurements with open-loop KPFM demonstrate that multiple force modulations may be applied at once. Further, this method can be applied to any tip-sample force that can be modulated, for example, electrostatic, magnetic, and photoinduced forces, showing its universality. Because HM enables in situ switching between high sensitivity and high spatial resolution with any periodic driving force, we foresee this technique as a critical advancement for multiscale functional imaging.


Effect of cooking locally available common bean (Obwelu) on iron and zinc retention, and pumpkin (Sweet cream) on provitamin A carotenoid retention in rural Uganda.
Pumpkin is a potential rich source of vitamin A precursors called provitamin A carotenoids (PVACs), while common bean is a potential rich source of iron and zinc. This study evaluated the effect of cooking locally available pumpkin, Sweet cream (Cucurbita moschata) on PVACs retention in Uganda. Furthermore, the effect of cooking locally available common bean, Obwelu (Phaseolus vulgaris) on iron and zinc retention was evaluated. Expert caregivers from the local community cooked pumpkin by either boiling or steaming, while common bean was cooked by either boiling with prior soaking or boiling without prior soaking. PVACs in raw and cooked pumpkin were analyzed by high-performance liquid chromatography (HPLC), while iron and zinc in raw and cooked common bean were analyzed by flame atomic absorption spectroscopy (FAAS). Conversion of PVACs into vitamin A retinol activity equivalents (RAE) was calculated using the Institute of Medicine (2001) recommendations for the bioconversion of PVACs into vitamin A. Micronutrient retention was measured using true retention. β-carotene, α-carotene, and vitamin A content in raw pumpkin was 1,704 µg/100 g, 46 µg/100 g and 1,437 µgRAE/100 g, respectively. Either boiling or steaming pumpkin resulted in over 100% retention of PVACs and vitamin A. Iron and zinc retention for boiled common bean with prior soaking was 92.2% and 91.3%, respectively. Boiling common bean without prior soaking resulted in 88.4% and 75.6% retention of iron and zinc, respectively. In conclusion, to retain a high proportion of PVACs caregivers should be advised to cook Sweet cream by either boiling or steaming, while to retain a high proportion of iron and zinc, Obwelu should be prepared by boiling with prior soaking.


[Intravenous Sedation and Repeated "the Same Day General Anesthesia" for a School-age Boy with Dandy-Walker Syndrome and Dentinogenesis].
Dandy-Walker syndrome (DWS) is characterized by perfect or partial defect of the cerebellum vermis and cystic dilatation of the posterior fossa communicating with the fourth ventricle. Common clinical signs are mental retardation, cerebellar ataxia, and those of increased intracranial pressure (ICP). Associated congenital anomalies are craniofacial, cardiac, renal, and skeletal abnormalities. We experienced a case of intravenous sedation and six times of "the same day" general anesthesia for a school-aged boy (10-13 years old) with DWS and hypodentinogenesis. The patient underwent an examination and dental treatments. We had to pay attention to airway management tracheal tube selection and control of ICP. In addition, we should prevent tooth injuries through mishaps during tracheal intubations, since all-tooth-hypoplasia with fragile dental crowns was strongly suggested in this case. Detailed postoperative care is also required for general anesthesia afflicted with DWS.


RipD (Rv1566c) from Mycobacterium tuberculosis: adaptation of an NlpC/p60 domain to a non-catalytic peptidoglycan-binding function.
Enzymes carrying NlpC/p60 domains, for instance RipA and RipB from Mycobacterium tuberculosis, are bacterial peptidoglycan hydrolases that cleave the peptide stems and contribute to cell wall remodelling during cell division. A member of this protein family, RipD (Rv1566c) from M. tuberculosis described in the present study, displays sequence alterations in the NlpC/p60 catalytic triad and carries a pentapeptide repeat at its C-terminus. Bioinformatics analysis revealed RipD-like proteins in eleven mycobacterial genomes, whereas similar pentapeptide repeats occur in cell-wall-localized bacterial proteins and in a mycobacteriophage. In contrast with previously known members of the NlpC/p60 family, RipD does not show peptidoglycan hydrolase activity, which is consistent with the sequence alterations at the catalytic site. A strong interaction of the catalytically inactive core domain with peptidoglycan is however retained, presenting the first example of the NlpC/p60 domains that evolved to a non-catalytic peptidoglycan-binding function. Full-length RipD carrying the C-terminal repeat shows, however, a decrease in binding affinity to peptidoglycan, suggesting that the C-terminal tail modulates the interaction with bacterial cell wall components. The pentapeptide repeat at the C-terminus does not adopt a defined secondary structure in solution which is in accordance with results from the 1.17 Å (1 Å=0.1 nm) crystal structure of the protein carrying two repeat units.


In vitro human cell line models to predict clinical response to anticancer drugs.
In vitro human cell line models have been widely used for cancer pharmacogenomic studies to predict clinical response, to help generate pharmacogenomic hypothesis for further testing, and to help identify novel mechanisms associated with variation in drug response. Among cell line model systems, immortalized cell lines such as Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) have been used most often to test the effect of germline genetic variation on drug efficacy and toxicity. Another model, especially in cancer research, uses cancer cell lines such as the NCI-60 panel. These models have been used mainly to determine the effect of somatic alterations on response to anticancer therapy. Even though these cell line model systems are very useful for initial screening, results from integrated analyses of multiple omics data and drug response phenotypes using cell line model systems still need to be confirmed by functional validation and mechanistic studies, as well as validation studies using clinical samples. Future models might include the use of patient-specific inducible pluripotent stem cells and the incorporation of 3D culture which could further optimize in vitro cell line models to improve their predictive validity.


Connected Component Analysis of Dynamical Perturbation Contact Networks.
Describing protein dynamical networks through amino acid contacts is a powerful way to analyze complex biomolecular systems. However, due to the size of the systems, identifying the relevant features of protein-weighted graphs can be a difficult task. To address this issue, we present the connected component analysis (CCA) approach that allows for fast, robust, and unbiased analysis of dynamical perturbation contact networks (DPCNs). We first illustrate the CCA method as applied to a prototypical allosteric enzyme, the imidazoleglycerol phosphate synthase (IGPS) enzyme from Thermotoga maritima bacteria. This approach was shown to outperform the clustering methods applied to DPCNs, which could not capture the propagation of the allosteric signal within the protein graph. On the other hand, CCA reduced the DPCN size, providing connected components that nicely describe the allosteric propagation of the signal from the effector to the active sites of the protein. By applying the CCA to the IGPS enzyme in different conditions, i.e., at high temperature and from another organism (yeast IGPS), and to a different enzyme, i.e., a protein kinase, we demonstrated how CCA of DPCNs is an effective and transferable tool that facilitates the analysis of protein-weighted networks.


First macrobiota biomineralization was environmentally triggered.
Why large and diverse skeletons first appeared ca 550 Ma is not well understood. Many Ediacaran skeletal biota show evidence of flexibility, and bear notably thin skeletal walls with simple, non-hierarchical microstructures of either aragonite or high-Mg calcite. We present evidence that the earliest skeletal macrobiota, found only in carbonate rocks, had close soft-bodied counterparts hosted in contemporary clastic rocks. This includes the calcareous discoidal fossil Suvorovella, similar to holdfasts of Ediacaran biota taxa previously known only as casts and moulds, as well as tubular and vase-shaped fossils. In sum, these probably represent taxa of diverse affinity including unicellular eukaryotes, total group cnidarians and problematica. Our findings support the assertion that the calcification was an independent and derived feature that appeared in diverse groups where an organic scaffold was the primitive character, which provided the framework for interactions between the extracellular matrix and mineral ions. We conclude that such skeletons may have been acquired with relative ease in the highly saturated, high alkalinity carbonate settings of the Ediacaran, where carbonate polymorph was further controlled by seawater chemistry. The trigger for Ediacaran biomineralization may have been either changing seawater Mg/Ca and/or increasing oxygen levels. By the Early Cambrian, however, biomineralization styles and the range of biominerals had significantly diversified, perhaps as an escalating defensive response to increasing predation pressure. Indeed skeletal hardparts had appeared in clastic settings by Cambrian Stage 1, suggesting independence from ambient seawater chemistry where genetic and molecular mechanisms controlled biomineralization and mineralogy had become evolutionarily constrained.


Imaging of tumor angiogenesis in rat brains in vivo by photoacoustic tomography.
Green laser pulses at a wavelength of 532 nm from a Q-switched Nd:YAG laser were employed as irradiation sources for photoacoustic tomography (PAT). The vascular structure of the brain was imaged clearly, with optimal contrast, because blood has strong absorption near this wavelength. The photoacoustic images of rat brain tumors in this study clearly reveal the angiogenesis that is associated with tumors. Brain tumors can be identified based on the distorted vascular architecture of brain tumorigenesis and related vascular changes, such as hemorrhage. This research demonstrates that PAT can potentially provide a powerful tool for small-animal biological research.


[Allogeneic bone marrow transplantation in children with severe acquired aplastic anemia. The long-term results. The Spanish Group for Bone Marrow Transplantation in Children (GETMON)].
The objective of this study was to evaluate retrospectively the efficacy of allogeneic BMT in the treatment of childhood severe acquired aplastic anemia (SAAA). Twenty-seven children aged 2 to 16 years (median 11 years) received a BMT from an HLA identical sibling. Conditioning consisted in irradiation (total, nodal or thoraco-abdominal) plus cyclophosphamide (120-200 mg/kg) in 15 patients and cyclophosphamide alone (200 mg/kg) in the rest. Prophylaxis for graft-versus-host disease (GVHD) was cyclosporine and methotrexate in most patients. Twenty-four children achieved the bone marrow graft at a median of 18 days (neutrophils) and 21 days (platelets). Two patients failed engraftment and 1 had a late graft rejection. Three patients developed acute GVHD grades 3-4 and six chronic GVHD, which was extensive in 4 of them. Twenty patients/71%) are alive and disease-free at a median follow-up of 110 months and the estimated disease free survival at 6 years is 67%. Our results confirm that allogeneic bone marrow transplantation from an HLA identical sibling is the best treatment modality for children with SAAA. Acute GVHD associated with infections and graft rejection were responsible for treatment failures.


Targeting multiple health risk behaviours among vocational education students using electronic feedback and online and telephone support: protocol for a cluster randomised trial.
Technical and Further Education (TAFE) colleges are the primary provider of vocational education in Australia. Most TAFE students are young adults, a period when health risk behaviours become established. Furthermore, high rates of smoking, risky alcohol consumption, inadequate fruit and vegetable intake and insufficient physical activity have been reported in TAFE students. There have been no intervention studies targeting multiple health risk behaviours simultaneously in this population. The proposed trial will examine the effectiveness of providing TAFE students with electronic feedback regarding health risk behaviours and referral to a suite of existing online and telephone services addressing smoking, risky alcohol consumption, fruit and vegetable intake, and physical activity levels. A two arm, parallel, cluster randomised trial will be conducted within TAFE campuses in New South Wales (NSW), Australia. TAFE classes will be randomly allocated to an intervention or control condition (50 classes per condition). To be eligible, students must be: enrolled in a course that runs for more than 6 months; aged 16 years or older; and not meet Australian health guideline recommendations for at least one of the following: smoking, alcohol consumption, fruit and/or vegetable intake, or physical activity. Students attending intervention classes, will undertake via a computer tablet a risk assessment for health risk behaviours, and for behaviours not meeting Australian guidelines be provided with electronic feedback about these behaviours and referral to evidence-based online programs and telephone services. Students in control classes will not receive any intervention. Primary outcome measures that will be assessed via online surveys at baseline and 6 months post-recruitment are: 1) daily tobacco smoking; 2) standard drinks of alcohol consumed per week; 3) serves of fruit consumed daily; 4) serves of vegetables consumed daily; and 5) metabolic equivalent minutes of physical activity per week. Proactive enrolment to existing online and telephone services has the potential to address modifiable determinants of disease. This trial will be the first to examine a potentially scalable intervention targeting multiple health risk behaviours among students in the vocational training setting. Australian New Zealand Clinical Trials Registry ACTRN12615000105549; Registered 5/2/15.


Stronger effects of Roundup than its active ingredient glyphosate in damselfly larvae.
Pesticides are causing strong decreases in aquatic biodiversity at concentrations assumed safe by legislation. One reason for the failing risk assessment may be strong differences in the toxicity of the active ingredient of pesticides and their commercial formulations. Sublethal effects, especially those on behaviour, have been largely ignored in this context, yet can be equally important as lethal effects at the population and ecosystem levels. Here, we compared the toxicity of the herbicide Roundup and its active ingredient glyphosate on survival, but also on ecologically relevant sublethal traits (life history, behaviour and physiology) in damselfly larvae. Roundup was more toxic than glyphosate with negative effects on survival, behaviour and most of the physiological traits being present at lower concentrations (food intake, escape swimming speed) or even only present (survival, sugar and total energy content and muscle mass) following Roundup exposure. This confirms the toxicity of the surfactant POEA. Notably, also glyphosate was not harmless: a realistic concentration of 2mg/l resulted in reduced growth rate, escape swimming speed and fat content. Our results therefore indicate that the toxicity of Roundup cannot be fully attributed to its surfactant, thereby suggesting that also the new generation of glyphosate-based herbicides with other mixtures of surfactants likely will have adverse effects on non-target aquatic organisms. Ecotoxicological studies comparing the toxicity of active ingredients and their commercial formulations typically ignore behaviour while the here observed differential effects on behaviour likely will negatively impact damselfly populations. Our data highlight that risk assessment of pesticides ignoring sublethal effects may contribute to the negative effects of pesticides on aquatic biodiversity.


Bovine factor v: a calcium-containing metalloprotein.
Although coagulation factor Xa requires Ca2+ for binding to phospholipid, factor V, the other protein component in the prothrombinase enzyme complex, binds tightly to phospholipid in the absence of Ca2+. To explore the possibility that calcium might be present in the fact V molecule, the effect of several chelators, including oxalate, citrate, pyrophosphate, and EDTA, on factor V activity has been studied. A time- and concentration-dependent inhibition of factor V which reflects the respective association constants for calcium of each chelator is observed. The inhibition can be prevented by the prior addition of calcium and manganese but not magnesium. Reversal of the activity loss can be accomplished at high protein concentrations by the addition of calcium, the removal of the chelator by gel filtration, or an increase in temperature. Factor V contains 1 g atom of calcium per 300,000 daltons which is not removed by incubation with EDTA under nondenaturing conditions. Thus, the inhibition by EDTA is due to binding to calcium associated with factor V. In 8 M urea, EDTA can remove over 80% of the calcium, demonstrating the importance of the native structure in maintaining the calcium binding site. Prior binding of phospholipids to factor V prevents inhibition by EDTA. The results suggest that phospholipids complex at the calcium site on the factor V metallopretein.


Cannabis-Based Medicine for Neuropathic Pain and Spasticity-A Multicenter, Randomized, Double-Blinded, Placebo-Controlled Trial.
Patients with multiple sclerosis (MS) and spinal cord injury (SCI) commonly sustain central neuropathic pain (NP) and spasticity. Despite a lack of consistent evidence, cannabis-based medicine (CBM) has been suggested as a supplement treatment. We aimed to investigate the effect of CBM on NP and spasticity in patients with MS or SCI. We performed a randomized, double-blinded, placebo-controlled trial in Denmark. Patients aged ≥18 years with NP (intensity >3, ≤9 on a numerical rating scale (NRS0-10) and/or spasticity (>3 on NRS0-10) were randomized to treatment consisting of either delta-9-tetrahydrocannabinol (THC), cannabidiol (CBD), a combination of THC&CBD in maximum doses of 22.5 mg, 45 mg and 22.5/45 mg per day, respectively, or placebo. A baseline registration was performed before randomization. Treatment duration was six weeks followed by a one-week phaseout. Primary endpoints were the intensity of patient-reported NP and/or spasticity. Between February 2019 and December 2021, 134 patients were randomized (MS n = 119, SCI n = 15), where 32 were assigned to THC, 31 to CBD, 31 to THC&CBD, and 40 to placebo. No significant difference was found for: mean pain intensity (THC 0.42 (-0.54-1.38), CBD 0.45 (-0.47-1.38) and THC&CBD 0.16 (-0.75-1.08)), mean spasticity intensity (THC 0.24 (-0.67-1.45), CBD 0.46 (-0.74-1.65), and THC&CBD 0.10 (-1.18-1.39), secondary outcomes (patient global impression of change and quality of life), or any tertiary outcomes. We aimed to include 448 patients in the trial; however, due to COVID-19 and recruitment challenges, fewer were included. Nevertheless, in this four-arm parallel trial, no effect was found between placebo and active treatment with THC or CBD alone or in combination on NP or spasticity in patients with either MS or SCI. The trial was registered with the EU Clinical Trials Register EudraCT (2018-002315-98).