Pharmacokinetics of puerarin in pregnant rats at different stages of gestation after oral administration.
This study aims to observe the effects of gestational stage on the pharmacokinetics of puerarin after oral administration in rats. The pharmacokinetics of puerarin was studied in pregnant rats using a sensitive and reproducible high-performance liquid chromatography/ultraviolet method. The concentration-time curves in both normal and pregnant rats were fit into a two-compartment model. The results indicated that gestation influences the pharmacokinetics of puerarin at different levels, especially during the early stages of pregnancy. Furthermore, puerarin penetrates the placental barrier and maintains high concentrations in fetal rat plasma. Therefore, puerarin administration should be carefully considered in pregnant women.


Enhanced Platelet Toll-like Receptor 2 and 4 Expression in Acute Coronary Syndrome and Stable Angina Pectoris.
Evidence is accumulating that toll-like receptors (TLR) are involved in the initiation and progression of cardiovascular disease. Enhanced expression of these receptors on monocytes has been shown in patients with acute coronary syndrome (ACS). However, expression on platelets in this group of patients has not been evaluated yet. We aimed to demonstrate the possible relationship of platelet TLR-2 and TLR-4 expressions with stable coronary artery disease and ACS pathogenesis. In this observational case-control study, 40 patients diagnosed with ACS (unstable angina pectoris, non-ST-segment elevation and ST-segment elevation ACS), 40 patients diagnosed with stable coronary artery disease, and 40 age- and gender-matched subjects with normal coronary arteries were involved. Platelet TLR-2 and TLR-4 expressions were evaluated by flow cytometry in peripheral venous blood samples obtained before coronary angiography. A total of 120 patients (60.7 ± 12.3 years, 50% men) were included. Median platelet TLR-2 and TLR-4 expressions were greater in patients with ACS compared to those with stable angina pectoris and normal coronary arteries (29.5% vs 10.5% vs 3.0%, p <0.001 and 40.5% vs 11.5% vs 3.0%, p <0.001, respectively). Median platelet TLR-2 and TLR-4 expressions were also greater in patients with stable angina pectoris compared to those with normal coronary arteries (p <0.05). In conclusion, this is the first study demonstrating enhanced TLR-2 and TLR-4 expressions on platelets in patients with ACS. These findings may suggest that platelet TLR expression as a novel potential prophylactic and therapeutic target in ACS.


Comparative evaluation of different methods for calculation of cerebral blood flow (CBF) in nonanesthetized rabbits.
The measurement of cerebral blood flow (CBF) by the extracranial detection of the radioactivity of 133Xe injected into an internal carotid artery has proved to be of considerable value for the investigation of cerebral circulation in conscious rabbits. Methods are described for calculating CBF from the curves of clearance of 133Xe, and include exponential analysis (two-component model), initial slope, and stochastic method. The different methods of curve analysis were compared in order to evaluate the fitness with the theoretical model. The initial slope and stochastic methods, compared with the biexponential model, underestimate the CBF by 35% and 46% respectively. Furthermore, the validity of recording the clearance curve for 10 min was tested by comparing these CBF values with those obtained from the whole curve. CBF values calculated with the shortened procedure are overestimated by 17%. A correlation exists between the "10 min" CBF values and the CBF calculated from the whole curve; in spite of that, the values are not accurate for limited animal populations or for single animals. The extent of the two main compartments into which the CBF is divided was also measured. There is no correlation between CBF values and the extent of the relative compartment. This fact suggests that these two parameters correspond to different biological entities.


Hypermethylation of the suppressor of cytokine signalling-1 (SOCS-1) in myelodysplastic syndrome.
Transcriptional silencing because of hypermethylation is now recognised to be a hallmark of human tumours. In contrast to acute myeloid leukaemia (AML), comparably little is known about aberrant methylation in myelodysplastic syndrome (MDS), a heterogeneous clonal stem cell disorder with a risk of transformation into secondary AML of up to 30%. Recent evidence demonstrates that suppressor of cytokine signalling SOCS-1, a negative regulator of cytokine pathways, may act as a tumour suppressor gene, and inactivation because of hypermethylation was shown in various malignancies. Employing a newly developed quantitative real-time polymerase chain reaction-based methylation assay we analysed, for the first time, SOCS-1 methylation in MDS and found disease-specific hypermethylation in 27 of 86 MDS patients (31%). Demethylation experiments provided direct evidence that aberrant methylation of SOCS-1 induces transcriptional silencing in myeloid cells. In addition, by analysing the expression of signal transducers and activators of transcription (STAT)-induced genes we provide for the first time evidence that the activity of the Janus kinase/STAT pathway is increased in primary patient samples showing SOCS-1 hypermethylation.


Aberrant promoter methylation of the vimentin gene may contribute to colorectal carcinogenesis: a meta-analysis.
This meta-analysis of published cohort studies was conducted to evaluate how closely the promoter methylation of the vimentin gene is correlated with the pathogenesis of colorectal carcinogenesis (CRC). The Web of Science (1945 ~ 2013), Cochrane Library Database (issue 12, 2013), PubMed (1966 ~ 2013), EMBASE (1980 ~ 2013), CINAHL (1982 ~ 2013), and Chinese Biomedical Database (CBM) (1982 ~ 2013) were searched without language restrictions. Meta-analyses were conducted using Stata software (Version 12.0, Stata Corporation, College Station, TX, USA). Odds ratios (ORs) and 95 % confidence intervals (95 %CI) were calculated. Seven clinical cohort studies with a total of 467 CRC subjects met our inclusion criteria. Our meta-analysis results demonstrated that the frequency of vimentin promoter methylation in cancer tissues was significantly higher than in normal and benign tissues (cancer tissues vs. normal tissues: OR = 32.41, 95 %CI = 21.04 ~ 49.93, P < 0.001; cancer tissues vs. benign tissues: OR = 1.60, 95 %CI 1.05 ~ 2.42, P = 0.028). Ethnicity-stratified analysis indicated that the frequency of aberrant vimentin promoter methylation was correlated with the pathogenesis of CRC in both Asians and Caucasians. The findings of our meta-analysis confirm that vimentin methylation may play a crucial role in the pathogenesis of CRC.


A summary of the BARREL campaigns: Technique for studying electron precipitation.
BARREL observed electron precipitation over wide range of energy and timescalesPrecipitating electron distribution is determined using spectroscopy for 19 January 2013 eventBARREL timing data has accuracy within sampling interval of 0.05 s.


Critical appraisal of physical rehabilitation measures after osteoporotic vertebral fracture.
Osteoporosis is asymptomatic until fracture occurs. Fracture of the vertebral bodies can be "silent" and is diagnosed incidentally on radiographic visualization. The occurrence of spontaneous vertebral fracture in an otherwise healthy individual is considered pathognomonic of spinal osteoporosis. Osteoporotic vertebral fractures and kyphotic posture are no longer disorders about which nothing can be done. Resistive training exercises can decrease the risk of vertebral fractures. Proprioception generated within joints, ligaments, and muscles contributes to awareness of the relative orientation of functional units of the spine at rest and in motion. This is fundamental to posture, balance, and locomotion. Proprioception reeducation can be utilized for improving posture and balance. The focus of this review is rehabilitative measures for management of vertebral fractures.


Synthesis of 3,5-bis(acrylaldehyde) boron-dipyrromethene and application in detection of cysteine and homocysteine in living cells.
Synthesis, characterization, and spectral and electrochemical properties of 3,5-bis(acrylaldehyde) BODIPY are described. The compound exhibited higher selectivity toward cysteine/homocysteine than toward other amino acids and thiol-containing compounds as shown by absorption and emission titration experiments and by experiments in living cells.


SETMAR, a case of primate co-opted genes: towards new perspectives.
We carry out a review of the history and biological activities of one domesticated gene in higher primates, SETMAR, by discussing current controversies. Our purpose is to open a new outlook that will serve as a framework for future work about SETMAR, possibly in the field of cognition development. What is newly important about SETMAR can be summarized as follows: (1) the whole protein sequence is under strong purifying pressure; (2) its role is to strengthen existing biological functions rather than to provide new ones; (3) it displays a tissue-specific pattern of expression, at least for the alternative-splicing it undergoes. Studies reported here demonstrate that SETMAR protein(s) may be involved in essential networks regulating replication, transcription and translation. Moreover, during embryogenesis, SETMAR appears to contribute to brain development. Our review underlines for the first time that SETMAR directly interacts with genes involved in brain functions related to vocalization and vocal learning. These findings pave the way for future works regarding SETMAR and the development of cognitive abilities in higher primates.


In Vitro Assay for Sensitive Determination of Human Blood PMN Responses.
Polymorphonuclear neutrophils (PMNs) are the most common leukocytes in the circulation and exhibit a wide range of distinct cellular phenomena as part of their microbicidal killing activities, including degranulation, phagocytosis, reactive oxygen species (ROS) production, adhesion, chemotaxis and production of PMN Extracellular Traps (NETs). As a simple in vitro test of PMN functional responses in human blood we have developed a multicolor flow cytometry-based assay of PMN cluster of differentiation (CD) surface marker expression. Short incubations of whole human blood can be performed in the presence of a wide range of agonists or inhibitors, followed by sensitive detection of changes in CD marker expression. This protocol has the advantage that small amounts of human blood are necessary and there are no PMN isolation steps, which can alter PMN activation status.


CT computer-optimized high-dose-rate brachytherapy with surface applicator technique for scar boost radiation after breast reconstruction surgery.
Immediate breast reconstruction has become increasingly prevalent after mastectomy for breast cancer. Postoperative scar boost radiation for the reconstructed breast presents many planning challenges due to the shape, size, and curvature of the scar. High-dose-rate (HDR) surface applicator brachytherapy is a novel and effective method of delivering scar boost radiation. Two cases, one with a saline implant and one with a transverse rectus abdominis musculocutaneous flap reconstruction, illustrate the method and advantages of HDR optimization of surface applicators. For 2 patients a mold of the breast was made with Aquaplast sheets. A reproducible system was used for arm positioning. Skin fiducials, including tattoos from external beam planning, were matched to fiducials on the mold. HDR catheters were sited on the mold at 1cm intervals, with the central catheter situated along the scar. Topographically, both scars demonstrated extreme curvature in both craniocaudal and mediolateral directions. A CT computer-optimized HDR plan was developed, with the reference dose prescribed at the skin surface. The dosimetry was compared to single-field and matched-field electron plans. This surface applicator technique provided a uniform skin dose of 100% to the entire clinical target volume (CTV) without hot spots in both patients. The patient position and surface applicator setup were consistently reproducible. The patients tolerated the treatment well with minimal skin erythema. In the single-field electron plan, skin dose was decreased to 50% at the periphery of the scar. Matching fields addressed this depth dose decrement, but resulted in large localized hot spots of more than 200% centrally in each field. CT computer-optimized HDR surface applicator brachytherapy provided a reproducible homogeneous method of treating highly curved scars on the reconstructed breast. Electron beam treatment would result in longer and more complex treatments yet still provide a less homogeneous dose than this surface applicator technique.


Combining non-canonical amino acid mutagenesis and native chemical ligation for multiply modifying proteins: A case study of α-synuclein post-translational modifications.
The Parkinson's disease associated protein α-synuclein (αS) has been found to contain numerous post-translational modifications (PTMs), in both physiological and pathological states. One PTM site of particular interest is serine 87, which is subject to both O-linked β-N-acetylglucosamine (gS) modification and phosphorylation (pS), with αS-pS87 enriched in Parkinson's disease. An often-overlooked aspect of these PTMs is their effect on the membrane-binding properties of αS, which are important to its role in regulating neurotransmitter release. Here, we show how one can study these effects by synthesizing αS constructs containing authentic PTMs and labels for single molecule fluorescence correlation spectroscopy measurements. We synthesize αS-gS87 and αS-pS87 by combining native chemical ligation with genetic code expansion approaches. We introduce the fluorophore by a click reaction with a non-canonical amino acid. Beyond the specific problem of PTM effects on αS, our studies highlight the value of this combination of methods for multiply modifying proteins.


Contrast-induced nephropathy in patients with renal insufficiency undergoing contrast-enhanced MDCT.
To evaluate the safety of contrast-enhanced MDCT in patients with renal impairment. We conducted a retrospective review of 938 patients with stable renal insufficiency (eGFR between 15 and 60 ml/min) who underwent contrast-enhanced MDCT. SCr levels were measured at baseline and 48-72 h after contrast medium administration. The incidence of contrast-induced nephropathy (CIN) in the total study population was assessed. As a control group, 1,164 separate patients with renal insufficiency who did not receive contrast medium for CT were also reviewed. The overall incidence of CIN in the patient population with renal insufficiency was 6.1 %; the incidence was 4.4 %, 10.5 % and 10.0 % for patients whose eGFR was 45-60, 30-45 and ≤30 ml/min, respectively (P < 0.01). In the control group, 5.8 % of patients showed an increase in the SCr level from the baseline. The increase in the SCr level showed no significant difference between the patients who received CM and those who did not (P = 0.82) CONCLUSIONS: The risk of CIN from contrast-enhanced MDCT in patients with renal insufficiency appeared to be low, and there was no significant difference in the incidence of CIN in comparison with patients who did not receive CM. • The contrast medium used for multidetector CT can induce nephropathy. • Contrast-induced nephropathy (CIN) developed in 6.1 % of patients with renal insufficiency. • However, nephropathy developed in 5.8 % of similar patients not receiving contrast medium. • Thus, the risk of CIN associated with MDCT appears to be low. • Special care should still be taken in patients with renal insufficiency.


Cloned cDNA of A/swine/Iowa/15/1930 internal genes as a candidate backbone for reverse genetics vaccine against influenza A viruses.
Reverse genetics viruses for influenza vaccine production usually utilize the internal genes of the egg-adapted A/Puerto Rico/8/34 (PR8) strain. This egg-adapted strain provides high production yield in embryonated eggs but does not necessarily give the best yield in mammalian cell culture. In order to generate a reverse genetics viral backbone that is well-adapted to high growth in mammalian cell culture, a swine influenza isolate A/swine/Iowa/15/30 (H1N1) (rg1930) that was shown to give high yield in Madin-Darby canine kidney (MDCK) cells was used as the internal gene donor for reverse genetics plasmids. In this report, the internal genes from rg1930 were used for construction of reverse genetics viruses carrying a cleavage site-modified hemagglutinin (HA) gene and neuraminidase (NA) gene from a highly pathogenic H5N1 virus. The resulting virus (rg1930H5N1) was low pathogenic in vivo. Inactivated rg1930H5N1 vaccine completely protected chickens from morbidity and mortality after challenge with highly pathogenic H5N1. Protective immunity was obtained when chickens were immunized with an inactivated vaccine consisting of at least 2(9) HA units of the rg1930H5N1 virus. In comparison to the PR8-based reverse genetics viruses carrying the same HA and NA genes from an H5N1 virus, rg1930 based viruses yielded higher viral titers in MDCK and Vero cells. In addition, the reverse genetics derived H3N2 and H5N2 viruses with the rg1930 backbone replicated in MDCK cells better than the cognate viruses with the rgPR8 backbone. It is concluded that this newly established reverse genetics backbone system could serve as a candidate for a master donor strain for development of inactivated influenza vaccines in cell-based systems.


Neurobehavioural responses of the newborn. The effect of different forms of maternal analgesia.
A neurobehavioural assessment made of infants within 4 hours of delivery revealed that infants born to mothers receiving pethidine and promazine showed significantly worse overall scores than babies of mothers receiving either no analgesia or a lumbar epidural using bupivacaine.


Diagnosis of Parkinson's disease from hand drawing utilizing hybrid models.
Parkinson's disease is a nervous system abnormality marked by decreased dopamine levels in the brain. Parkinson's disease inhibits one's ability to move. Speech difficulty, changes in movement and handwriting, and other symptoms are common with Parkinson's disease. A collection of hand drawings is employed to predict Parkinson's disease. There are 102 spiral images in the hand drawing dataset. Due to the minimal size of the dataset, augmentation is utilized to increase it. After that, the augmented images are utilized to train several machine learning and deep learning models, as well as pre-trained networks like RESNET50, VGG16, AlexNet, and VGG19. The performance metrics of hybrid models of deep learning with machine learning and hybrid models of deep learning (for feature extraction) with deep learning (for classification) are then compared. It was observed that the hybrid model of RESNET-50 and SVM performed well with better performance measures compared to other Machine Learning, Deep Learning and Hybrid Models with an accuracy score of 98.45%, sensitivity score of 0.99 and specificity score of 0.98.


Optimized expression of classical swine fever virus E2 protein via combined strategy in Pichia pastoris.
Precaution of classical swine fever (CSF) is an important mission for the worldwide swine industry. Glycoprotein E2 is the leading antigen candidate for subunit vaccine of classical swine fever virus (CSFV). In this study, two Spy-tagged E2 genes were synthesized in vitro and subcloned into pMCO-AOX vector for intracellular expression in Pichia pastoris after methanol induction. Western blot analysis and semi-quantitative analysis showed that the yield of recombinant E2 protein was improved 17.87 folds by using co-translocational signal peptide cSIG. After the construction of the tandem multiple copy expression vectors, further increase of E2 production was observed by repetitive transforming expression vectors into P. pastoris genome. Finally, the yeast transformants harboring 8 or 16 copies of cSIG-E2-Spy increased the E2 expression level by 27.01-fold or 30.72-fold, respectively. These results demonstrate that utilizing co-translocational signal peptide together with multi-copy integration strategy can increase the production of recombinant E2 protein efficiently.


1p/19q co-deletion status is associated with distinct tumor-associated macrophage infiltration in IDH mutated lower-grade gliomas.
Tumor-associated macrophages (TAM)s are critical regulators of glioma progression. As yet, however, TAMs in isocitrate dehydrogenase (IDH) mutated lower-grade gliomas (LGGs) have not been thoroughly investigated. The aim of this study was to determine whether 1p/19q co-deletion status affects the TAM phenotype or its prevalence in IDH mutated LGGs. TAMs in IDH mutated LGGs were analyzed using transcriptome data from 230 samples in the TCGA database in combination with transcriptome data from single-cell RNA sequencing of IDH-mutated LGGs. Proteins potentially involved in TAM regulation were examined by immuno-staining in primary LGG samples harboring IDH mutations. Essential signaling pathways regulating TAM phenotypes were investigated in a glioma mouse model using small molecule inhibitors. Most of the TAMs in IDH-mutated LGGs expressed the M1 activation markers CD86 and TNF, whereas a subset of individual TAMs co-expressed both M1 and M2-related markers. Bioinformatics analysis in combination with immuno-staining of IDH-mutated patient samples revealed higher amounts of TAMs expressing M2-related markers in 1p/19q non-codeletion IDH-mutated LGGs compared to 1p/19q codeletion LGGs. The levels of transforming growth factor beta 1 (TGFβ1) and macrophage colony-stimulating factor (M-CSF) were significantly higher in 1p/19q non-codeletion LGGs than in 1p/19q codeletion LGGs. M-CSF and TGFβ1 signal inhibition decreased tumor growth and modulated the TAM phenotype in a glioma mouse model. Our data indicate that 1p/19q co-deletion status relates to distinct TAM infiltration in gliomas, which is likely mediated by M-CSF and TGFβ1 signaling. M-CSF and TGFβ1 signaling may play a pivotal role in regulating the TAM phenotype in glioma.


Palliative Communication in the Pediatric Intensive Care Unit.
Communication is a central aspect of nursing care and is especially important when pertaining to progressive illnesses and end of life. This article reviews basic palliative care terminology and outlines a variety of communication frameworks from the "dos" to the "don'ts." These communication strategies are meant to be added to the nurse's "toolbox" so that nurses may use them in various scenarios. These communication tools are meant to help mitigate the stress and discomfort nurses often feel when using palliative communication or delivering bad news.


CircFOXO3 protects against osteoarthritis by targeting its parental gene FOXO3 and activating PI3K/AKT-mediated autophagy.
Osteoarthritis (OA) is a degenerative joint disorder causing pain and functional disability. Emerging evidence reveals that circular RNAs (circRNAs) play essential roles in OA progression and development. This study aimed to investigate the role of a novel circRNA factor, circFOXO3, in the progression of OA and elucidate its underlying molecular mechanism. The function of circFOXO3 in OA and interaction between circFOXO3 and its downstream mRNA target, forkhead box O3 (FOXO3), were evaluated by western blot (WB), immunofluorescence (IF), RNA immunoprecipitation, reverse transcription-quantitative PCR (RT-qPCR), and fluorescence in situ hybridization (FISH). Upregulation of circFOXO3 and autophagic flux were detected both in vivo and in vitro by WB, transmission electron microscopy (TEM), IF, and immunohistochemistry (IHC). A mouse model of OA was also used to confirm the role of circFOXO3 in OA pathogenesis in vivo. Decreased expression of circFOXO3 in OA cartilage tissues was directly associated with excessive apoptosis and imbalance between anabolic and catabolic factors of the extracellular matrix (ECM). Mechanistically, circFOXO3 functioned in cartilage by targeting its parental gene FOXO3 and activating autophagy. Intra-articular injection of lentivirus-circFOXO3 alleviated OA in the mouse model. In conclusion, our results reveal the key role played by circFOXO3 in OA progression; circFOXO3 overexpression may alleviate apoptosis of chondrocytes and promote anabolism of the ECM via activation of FOXO3 and autophagy, providing a potentially effective novel therapeutic strategy for OA.


Significance of duon mutations in cancer genomes.
Functional mutations in coding regions not only affect the structure and function of the protein products, but may also modulate their expression in some cases. This class of mutations, recently dubbed "duon mutations" due to their dual roles, can potentially have major impacts on downstream pathways. However their significance in diseases such as cancer remain unclear. In a survey covering 4606 samples from 19 cancer types, and integrating allelic expression, overall mRNA expression, regulatory motif perturbation, and chromatin signatures in one composite index called REDACT score, we identified potential duon mutations. Several such mutations are detected in known cancer genes in multiple cancer types. For instance a potential duon mutation in TP53 is associated with increased expression of the mutant allelic gene copy, thereby possibly amplifying the functional effects on the downstream pathways. Another potential duon mutation in SF3B1 is associated with abnormal splicing and changes in angiogenesis and matrix degradation related pathways. Our findings emphasize the need to interrogate the mutations in coding regions beyond their obvious effects on protein structures.


Turnover Rates and Factors Influencing Turnover of Korean Acute Care Hospital Nurses: A Retrospective Study Based on Survival Analysis.
This study aimed to explore turnover rates for Korean acute care hospital nurses and identify factors influencing their turnover. The study was retrospective in nature. Nurse cohort data were obtained from hospital status data from Korea's Health Insurance Review Assessment Service. The observation period was from January 1, 2012 to December 31, 2016, and data for 96,158 nurses were analyzed. Independent variables included nurses' age and sex and hospital setting, type, ownership, and nurse staffing level. Kaplan-Meier analysis was performed to estimate survival curves, and factors influencing turnover were analyzed using Cox's proportional hazard regression. The cumulative turnover probability for all nurses was .17, .29, .38, .45, and .50 for the first, second, third, fourth, and fifth years, respectively. The results showed that the longer the career duration, the lower the turnover rates. According to the factors influencing nurse turnover, both nurses' (i.e., sex and career duration) and hospitals' (i.e., hospital setting, type, ownership, and nurse staffing level) characteristics were statistically significant. It should be noted that the turnover rate of nurses with less than three year of career duration and of those with less than one year has been shown to be quite high. Therefore, target populations for acute care hospital nurse turnover should be expanded from new graduate nurses to experienced nurses with less than 3 years of career. Further studies are required to examine the causes of high turnover rates in hospitals that are small and/or have low nurse staffing levels.


Isolation of Human Neutrophils from Venous Blood.
Venous blood provides a ready source of large numbers of unstimulated granulocytes and mononuclear cells. Exploiting the differences in the relative densities of the leukocytes circulating in venous blood, one can separate leukocytes from erythrocytes as well as isolate the individual leukocyte populations in high purity for use in ex vivo studies. For selected functional studies, such as transcriptional analysis or cytokine quantitation, addition of an immunomagnetic negative selection step to the standard isolation protocol can yield highly purified human neutrophils.


Impact of Pregnancy on Weight Loss After Endoscopic Sleeve Gastroplasty.
Obesity and pregnancy are strictly related: on the one hand, obesity-one of the most common comorbidities in women of reproductive age-contributes to infertility and obesity-related pregnancy complications, whereas pregnancy is a condition in which, physiologically, the pregnant woman undergoes weight gain. Endoscopic sleeve gastroplasty (ESG) may be used for the treatment of obesity in women of childbearing age. A retrospective analysis was conducted to evaluate weight trajectories, the evolution of obesity-related comorbidities, and lifestyle modification in women who became pregnant after ESG. A comparison was made between childbearing-age women who became pregnant after ESG and non-pregnant women. A total of 150 childbearing-age women underwent ESG at a large tertiary medical center. Of these, 11 patients (33.4 ± 6.2 years) became pregnant after the procedure, following a mean time interval of 5.5 ± 3.9 months. Three women (two affected by polycystic ovary syndrome) reported difficulty getting pregnant before undergoing ESG. The mean preconception BMI was 31.9±4.0 kg/m2 (-7.24 ± 4.0 kg/m2 after ESG). Total body weight loss (TBWL, %) was 18.08 ± 8.00, 11.00 ± 11.08, and 12.08 ± 8.49, at the beginning of pregnancy, at the delivery, and at the first follow-up (19.6 ± 7.8 months after ESG). TBWL of at least 5% was achieved before pregnancy in all patients (73% reached a TBWL ≥ 10%). No significant differences in weight loss and QoL were found between the pregnancy and non-pregnancy groups up to 24 months after ESG. Endoscopic sleeve gastroplasty allows for adequate weight loss before and after pregnancy in patients with obesity.


Cooperative stimulation of specific gene transcription by epidermal growth factor and transforming growth factor type beta 1.
Transforming growth factor type beta 1 (TGF-beta 1) is a pleiotropic regulator of cell growth and differentiation which can potentiate or otherwise modify cellular responses to different growth factors such as epidermal growth factor (EGF). Since cellular responses to peptide growth factors are mediated, in part, through the regulation of specific gene expression, we have studied the effect of TGF-beta 1 on the EGF-dependent transcriptional of a diverse panel of genes which included the fibronectin gene, the cytoskeletal beta- and gamma-actin genes, and the c-fos protooncogene. The addition of a combination of TGF-beta 1 (10 ng/ml) and EGF (10 ng/ml) to quiescent AKR-2B cells maintained in serum-free medium resulted in a strong synergistic stimulation of transcription which was not evident using either growth factor individually at concentrations of 20-100 ng/ml. This effect was evident within 10 min and did not require the continual presence of TGF-beta 1 in the culture medium. Increased responsiveness of some gene promoters to EGF stimulation was evident for up to 6 h following a brief 10-min exposure to TGF-beta 1. Similar exposure to either platelet-derived growth factor or fibroblast growth factor had little or no effect on EGF-stimulated transcription. These results indicate that the transcriptional response of specific genes to EGF can be stably altered as a consequence of exposure to TGF-beta 1.


Quantitative ultrasound bone measurements: normal values and comparison with bone mineral density by dual X-ray absorptiometry.
Normative data for qualitative ultrasound (QUS) measurements: speed of sound (SOS), broadband ultrasound attenuation (BUA), and stiffness were established in 118 healthy women aged 20-86 years and in 42 healthy men aged 22-76 years. The relations between age, weight, height, and QUS were studied. QUS measurements were negatively correlated with age in both sexes. In women, age was accepted as first factor (R2 = 0.39 for SOS, 0.35 for BUA, and 0.45 for stiffness, P < 0.001); weight was accepted as second factor for BUA (R2 = 0.44, P < 0.001). In men, age was the only significant parameter (R2 = 0.41 for SOS, 0.39 for BUA, 0.43 for stiffness, P < 0.001). QUS measurements of the right and left feet were highly correlated unless unilateral foot pathology such as algodystrophy was present. Significant correlations were found between QUS of the calcaneus and dual X-ray absorptiometry (DXA) of the lumbar spine (R = 0.67, P < 0.01 for SOS; R = 0.57, P < 0.02 for BUA; R = 0.65, P < 0.01 for stiffness).


Hemidesmus indicus L. Br.: critical assessment of in vitro biotechnological advancements and perspectives.
Hemidesmus indicus (L.) R. Br. ex Schult., commonly known as Indian sarsaparilla or Anantamul, is an ethnopharmacologically important medicinal plant from the family Apocynaceae. Biosynthesis of an array of bioactive compounds such as methoxybenzaldehydes and their derivatives has been accountable for its intensive medicinal attributes. Low seed setting, over-exploitation from natural habitat and a need for selection and preservation of elite germplasms yielding higher levels of bioactive compounds have led researchers to enquire the standardization of micropropagation techniques and quantitative estimation of phytochemicals from H. indicus. The present work aims to have a comprehensive account on micropropagation in H. indicus, a comparative estimation of biotic and abiotic factors, viz. elicitors and precursors playing a crucial role in H. indicus tissue culture, a critical appraisal of applied protocols to nullify embellished claims and a discussion on future perspectives. The review also highlights the comparative effect of different plant growth regulators as well as their combined role in rhizogenesis, callogenesis, base callus formation, callusing and somatic embryo-mediated indirect organogenesis, outcome of explant selection, contribution of abiotic (temperature, photoperiod, moisture, inorganic molecules) factors, role of carbon source, application of sterilization techniques and associated success rates, role of factors essential for acclimatization and secondary metabolite production, synthetic seed production and genetic transformation techniques. The compiled information along with individual study details, respective outcome and variability of reports will aid in proper assessment of cited standardized tissue culture protocols in H. indicus especially in relation to secondary metabolite production. Moreover, the assessment of applicability of different methodologies in this aspect will aid in the selection of high-yielding germplasms or chemotypes which is not only profitable for industrial application but also important for basic and applied preclinical and clinical research studies. KEY POINTS: • Critical and updated assessment on in vitro biotechnology in Hemidesmus indicus. • Biotechnological advancement via Agrobacterium-mediated transformation. • Key shortcomings and future research directions Graphical abstract Effect of biotic and abiotic factors on Micropropagation and secondary metabolite synthesis in Hemidesmus indicus.


X-ray features to predict ankle fracture mechanism.
The genetic Lauge-Hansen classification has been traditionally used for reconstruction of the mechanism of ankle injury. The ability of the Lauge-Hansen classification to predict actual mechanism of the injury has been questioned in recent studies, leaving a void in medicolegal reasoning. The aim of this study is to identify morphologic features of malleolar fractures on plain X-rays that may be used to reveal the fracture mechanism. Radiographs of 78 patients with acute malleolar fractures were analyzed and compared with fracture mechanisms reported by these patients. A modified Pankovich classification of medial malleolus fractures and the presence of a posterior malleolus fracture were able to significantly predict the mechanism of fracture reported by the patient (p<0.05). Lateral fracture morphology was not useful for predicting the fracture mechanism except for infrasyndesmotic fractures pointing to a supination mechanism. A supination mechanism of the fracture can be predicted by observing pure ligamentous or chip-avulsion deltoid injury. The presence of an anterior colliculus fracture of the medial malleolus is correlated with a pronation mechanism. A fracture of the posterior malleolus correlates with pronation while an intact posterior malleolus correlateds with supination as a fracture mechanism. Absolute determination of fracture mechanism from the X-ray morphology of the fracture is impossible in the majority of cases.


Multi-layer dressing made of laminated electrospun nanowebs and cellulose-based adhesive for comprehensive wound care.
In this work, multi-layer wound dressing was made of laminated layers of electrospun fibers supported by adhesive sheet. Graft copolymerization of methyl methacrylate (MMA) and 2-Ethyl-1-hexyl acrylate (EHA) onto carboxymethyl cellulose (CMC) was conducted to obtain an adhesive sheet with 1.52 (N/cm2) loop tack, 1.7 (N/cm) peel strength and 25 s shear strength. Diclofenac sodium, anti-inflammatory drug, was loaded to the adhesive sheet with encapsulation efficiency 73%. The contact layer to wound was made of synthesized anti-bleeding agents, chitosan iodoacetamide (CI) loaded into electrospun polyvinyl alcohol (PVA) fibers. It was fabricated from fiber diameter 300 nm by electrospinning of 5% wt/v of CI (D.S. 18.7%) mixed with 10% wt/v PVA, at 20 kV and 17 cm airgap. The second, pain-relief layer was fabricated by encapsulating up to 50% wt/wt of capsaicin into gelatin nanofibers (197 nm) crosslinked by glyoxal. The third, antimicrobial layer was fabricated from PVA electrospun fibers loaded with 2% wt/wt gentamicin. Biocompatibility test showed insignificant adverse effects of the fabricated layers on fibroblast cells. Animal test on rat showed accelerated wound healing from 21 to 7 days for the multi-layer dressing. Histopathological findings corroborated the intactness of the epidermis layer of the treated samples.


Bacterial short-chain fatty acids and mucosal diseases of the colon.
This review examines the potential association between short-chain fatty acids and mucosal diseases of the colon, and discusses the implication of these relationships and the surgical management of large bowel disorders.


Turnip crinkle virus infection from RNA synthesized in vitro.
Genome-length cDNA clones of turnip crinkle virus (TCV) were constructed with SmaI and XbaI restriction sites engineered at the 5' and 3' termini, respectively. The genome-length cDNAs were positioned downstream of modified lambda and T7 phage promoters such that in vitro transcription resulted in RNAs with 5 extra nucleotides at the 3' end, and 1, 2, or 14 extra nucleotides at the 5' end depending on the construction. Transcripts with 14 extraviral 5' nucleotides were not infectious, while transcripts with 1 or 2 additional 5' nucleotides, with or without 5'-cap analog included in transcription reactions, were biologically active. These were approximately an order of magnitude less infectious than RNA extracted from TCV virions. The additional 5' nucleotides were not maintained in progeny viral RNAs isolated from plants.


Methods to Study Integrin Functions on Exosomes.
Exosomes represent an important group of extracellular vesicles. They are formed in endosomal compartments and are actively secreted to extracellular spaces. Several membrane proteins, including integrins, are present on the surface of exosomes. As exosomal integrins are competent for binding to ligand, they can play important roles in directing the tissue distribution of exosomes. Integrin-directed exosomal trafficking in vivo is involved in regulating the remodeling of cell homing niches for metastatic cancers and migrating lymphocytes. This chapter describes the methods used to study integrin functions on exosomes including: isolation and biophysical characterization of exosomes, exosomal integrin-ligand binding assays, and in vivo competitive exosome homing assays.


Highly sensitive tITP-CZE determination of l-histidine and creatinine in human blood plasma using field-amplified sample injection with mobility-boost effect.
2D computer simulation revealed that amino acids and weak electrolytes were cationized because of the migration of counter-ion from a BGE zone to a sample zone, which encouraged electrokinetic injection (EKI) of these analytes (by the mobility-boost (MB) effect). To investigate the effects of kinds and concentrations of counter-ions on the MB effect and the analyte amount injected into the capillary, experiments, and 1D computer simulations were performed. When acetate was used as the counter-ion, the LODs (S/N = 3) of l-histidine and creatinine, respectively, reached 0.10 and 0.25 nM because of the concentration effect by transient ITP (tITP). The concentrations of l-histidine and creatinine in human blood plasma obtained using the proposed method were agreed with those obtained using the conventional methods. The proposed method can be applied to the analysis of amino acids and weak bases that have similar pI and pKa to l-histidine and creatinine.


miR-543 promotes gastric cancer cell proliferation by targeting SIRT1.
SIRT1, a class III histone deacetylase, exerts inhibitory effects on tumorigenesis and is downregulated in gastric cancer. However, the role of microRNAs in the regulation of SIRT1 in gastric cancer is still largely unknown. Here, we identified miR-543 as a predicted upstream regulator of SIRT1 using 3 different bioinformatics databases. Mimics of miR-543 significantly inhibited the expression of SIRT1, whereas an inhibitor of miR-543 increased SIRT1 expression. MiR-543 directly targeted the 3'-UTR of SIRT1, and both of the two binding sites contributed to the inhibitory effects. In gastric epithelium-derived cell lines, miR-543 promoted cell proliferation and cell cycle progression, and overexpression of SIRT1 rescued the above effects of miR-543. The inhibitory effects of miR-543 on SIRT1 were also validated using clinical gastric cancer samples. Moreover, we found that miR-543 expression was positively associated with tumor size, clinical grade, TNM stage and lymph node metastasis in gastric cancer patients. Our results identify a new regulatory mechanism of miR-543 on SIRT1 expression in gastric cancer, and raise the possibility that the miR-543/SIRT1 pathway may serve as a potential target for the treatment of gastric cancer.


Neuroanatomic basis of impaired self-awareness after traumatic brain injury: findings from early computed tomography.
Impaired self-awareness (ISA) is common among patients with severe traumatic brain injury (TBI) and contributes to poorer functional outcome. There is keen interest in improving the understanding of this disorder as the neuroanatomic substrate of posttraumatic ISA is poorly understood. Determine whether (1) greater number of brain lesions, (2) greater volume of right hemisphere lesions, or (3) greater volume of frontal lesions is associated with greater levels of ISA after TBI. Prospective, observational study. Ninety-one TBI admissions to one of 2 National Institute on Disability and Rehabilitation Research TBI Model System (TBIMS) programs. Subjects met TBIMS inclusion criteria plus (1) resolution of posttraumatic amnesia (PTA) prior to rehabilitation discharge and (2) initial postinjury computerized tomography (CT) scan available as a hard copy and as an electronic file. CT scan lesions outlined by a board-certified neuroradiologist were measured using NIH Image, and resulting calculated lesion volumes/scan variables compared against demographic characteristics, TBI severity variables, and ISA variables measured by the Awareness Questionnaire (AQ) at the time of PTA resolution. Most subjects (78%) had at least 1 lesion on emergent CT, and contusion volumes varied in all regions of interest. Patients rated their functioning as more intact on the AQ than ratings of treating clinicians, consistent with ISA. Greater injury severity was associated with a greater degree of ISA. Multivariable linear regression revealed that, after adjustment for other predictors, the number of brain lesions was predictive of degree of ISA. Right hemisphere contusion or frontal lobe contusion volumes, however, were not predictive of degree of ISA. ISA was significantly associated with the number, but not with location or volume of focal lesions early after TBI. Posttraumatic ISA may reflect disruption in the integrated operation of broadly distributed neural networks, with lesion burden in any specific region being less relevant than disruption across multiple regions. Further imaging research is warranted to confirm these findings and to provide insight into the distributed networks required for self-awareness.


Cisplatin, vindesine and bleomycin (CVB) combination chemotherapy of advanced non-small cell lung cancer.
Fifty-four patients with advanced squamous, large-cell or adeno-carcinoma of the lung were treated with a three drug regimen of high-dose cisplatin, vindesine and a four-day bleomycin infusion (CVB). All patients had measurable disease; none had previously received chemotherapy. Of 52 patients evaluable for response, 20 (38%) had complete or partial remissions. The median duration of remission was 8 months (range, 6 1/2-19+ months), with eight patients continuing in remission. The median duration of survival for responding patients was 16 months versus five months for nonresponding patients (P greater than 0.001). Toxicity included mild to moderate myelosuppression, peripheral neuropathy, and nephrotoxicity, and was generally manageable. The addition of a four-day bleomycin infusion did not appear to offer a significant advantage over the combination of high-dose cisplatin and vindesine in the treatment of NSCLC. The response rate and survival duration produced by the CVB protocol were similar to those reported for the two-drug combination of vindesine and high-dose cisplatin.


Establishing an untargeted-to-MRM liquid chromatography-mass spectrometry method for discriminating reconstituted milk from ultra-high temperature milk.
The counterfeit use of reconstituted milk to simulate fresh milk in some countries is largely driven by profiteering; hence, establishing a robust market-surveillance method is an important objective. In this study, an untargeted metabolomics approach that uses ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was established to discover biomarkers that can be used to classify UHT and reconstituted milk. Furthermore, 36 positive- and 24 negative-ionization mode features were selected as candidate markers to establish a UPLC-tandem mass spectrometry (UPLC-MS/MS) multiple reaction monitoring (MRM) method. The support vector machine (SVM) method was used to process the responses of the selected potential markers, and 100% classifiability was observed. The marker metabolites could be divided into three categories by hierarchical clustering analysis: peptides, lipids, and nucleic acids. The results suggest that the untargeted-to-MRM metabolomics method is an effective tool for distinguishing between UHT and reconstituted milk.


C6 modification of the pyridinone core of thrombin inhibitor L-374,087 as a means of enhancing its oral absorption.
1 (L-374,087) is a potent, selective, efficacious, and orally bioavailable thrombin inhibitor that contains a core 3-amino-2-pyridinone moiety. Replacement of the C6 pyridinone methyl group of 1 by a propyl group gave 5 (L-375,052), which retained all the excellent properties of 1, and also yielded higher plasma levels after oral dosing in dogs and rats.


The elevated T-maze as an experimental model of anxiety.
Anxiety disorders are heterogeneous and existing animal models do not discriminate specific types of anxiety. The elevated T-maze is being developed to fulfill this purpose. The apparatus consists of three elevated arms, one enclosed and two open. Inhibitory avoidance--representing learned fear--is measured by recording the time taken to leave the enclosed arm in three consecutive trials. Unconditioned fear is evaluated by recording the time to escape from the open arm. Restraining the animals at the end of the enclosed arm for 30 s did not change the first (baseline) withdrawal latency, indicating that rats are not escaping from the experimenter's hand. In addition, rats trained in a T-maze with the three arms enclosed did not show the usual increase in withdrawal latency over the three consecutive trials. These results indicate that open arm experience, not handling, motivates inhibitory avoidance learning. The same experiment also showed that the latency to leave the open arm did not undergo habituation over five consecutive trials, thereby providing evidence of an aversive motivation for this response. The anxiolytic agents diazepam (benzodiazepine), buspirone and ipsapirone (5-HT1A agonists) as well as ritanserin (5-HT2 antagonist) selectively impaired inhibitory avoidance while leaving one-way escape unchanged. Similar results were obtained with three putative anxiolytics: the 5-HT2B/2C antagonists SB 200646A and SER 082, and the 5-HT2A antagonist SR 46349B. However, RP 62203, another 5-HT2A antagonist, was ineffective on both tasks. In contrast to the above anxiolytics, the anxiogenic agents yohimbine, TFPP and mCPP facilitated inhibitory avoidance. Escape was not affected by yohimbine, but was moderately attenuated by the two 5-HT2C/2B agonists. The 5-HT releaser and uptake inhibitor D-fenfluramine tended to enhance inhibitory avoidance, while impairing one-way escape in a dose-dependent way. The antidepressant clomipramine also had an anxiogenic-like effect on inhibitory avoidance, but did not affect escape from the open arm. Conversely, the phenethylamine hallucinogen ALEPH 2 did not affect inhibitory avoidance while impairing escape. Nevertheless, the similar compound and 5-HT2A agonist DOI was devoid of any effect. Also ineffective were the psychomotor stimulants D,L-amphetamine and caffeine, the reversible monoaminoxidase-A inhibitor moclobemide and the neuroleptic haloperidol. Finally, micro-injection into the dorsal raphe nucleus of two drugs that stimulate 5-HT neurons, the excitatory amino acid kainic acid and the benzodiazepine inverse agonist FG 7142, facilitated inhibitory avoidance. Kainate also significantly impaired escape. In contrast, intra-raphe 8-OH-DPAT, which inhibits 5-HT neurons, selectively impaired inhibitory avoidance in a manner similar to systemically administered anxiolytics. These behavioral and pharmacological results support the view that inhibitory avoidance in the elevated T-maze may be related to generalized anxiety disorder, while one-way escape may be associated with panic disorder.


Resveratrol preserves mitochondrial function in a human post-mitotic cell model.
Dysfunctions caused by genetic defects in the mitochondrial DNA (mtDNA) of humans are called mitochondrial diseases; however, mtDNA mutations are also associated with aging and age-related diseases. Here, we present an original cellular model that allows gathering information on molecules that might contrast or prevent mitochondrial dysfunctions and their related diseases. This model allowed us to show that resveratrol (RSV), a phytochemical present in food, exerts protective effects at low concentrations on resting human fibroblasts carrying dysfunctional respiratory chain Complex I. Cells were maintained both in resting condition, to mimic the high energy demanding post-mitotic tissues (serum absence and gramicidin presence), and under glucose deficiency to push the synthesis of ATP via oxidative phosphorylation. Pre-incubation with RSV prolonged the viability of the fibroblasts exposed to rotenone, a well-known specific inhibitor of the respiratory chain Complex I, and decreased mitochondrial fragmentation. It significantly prevented the oxidative phosphorylation impairment indirectly caused by the rotenone-mediated Complex I inhibition, allowing for an almost complete preservation of the cellular ATP level. Indeed, RSV limited the rotenone-induced reactive oxygen species increase, allowing for the maintenance of a functional mitochondrial membrane potential. These findings indicate the potential usage of resveratrol to prevent or possibly treat many disorders, in which the bioenergetic defects and oxidative stress are the primary (mitochondrial encephalomyopathy), or the secondary (age-related diseases) causes of the pathology; and to also assist cell senescence during aging.


Apoptosis and pathogenesis of melanoma and nonmelanoma skin cancer.
Skin cancers, i.e., basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and melanoma, belong to the most frequent tumors. Their formation is based on constitutional and/or inherited factors usually combined with environmental factors, mainly UV-irradiation through long term sun exposure. UV-light can randomly induce DNA damage in keratinocytes, but it can also mutate genes essential for control and surveillance in the skin epidermis. Various repair and safety mechanisms exist to maintain the integrity of the skin epidermis. For example, UV-light damaged DNA is repaired and if this is not possible, the DNA damaged cells are eliminated by apoptosis (sunburn cells). This occurs under the control of the p53 suppressor gene. Fas-ligand (FasL), a member of the tumor necrosis superfamily, which is preferentially expressed in the basal layer of the skin epidermis, is a key surveillance molecule involved in the elimination of sunburn cells, but also in the prevention of cell transformation. However, UV light exposure downregulates FasL expression in keratinocytes and melanocytes leading to the loss of its sensor function. This increases the risk that transformed cells are not eliminated anymore. Moreover, important control and surveillance genes can also be directly affected by UV-light. Mutation in the p53 gene is the starting point for the formation of SCC and some forms of BCC. Other BCCs originate through UV light mediated mutations of genes of the hedgehog signaling pathway which are essential for the maintainance of cell growth and differentiation. The transcription factor Gli2 plays a key role within this pathway, indeed, Gli2 is responsible for the marked apoptosis resistance of the BCCs. The formation of malignant melanoma is very complex. Melanocytes form nevi and from the nevi melanoma can develop through mutations in various genes. Once the keratinocytes or melanocytes have been transformed they re-express FasL which may allow the expanding tumor to evade the attack of immune effector cells. FasL which is involved in immune evasion or genes which govern the apoptosis resistance, e.g., Gli2 could therefore be prime targets to prevent tumor formation and growth. Attempts to silence these genes by RNA interference using gene specific short interfering RNAs (siRNAs) or short hairpin RNAs (shRNAs) have been functionally successful not only in tissue cultures and tumor tissues, but also in a mouse model. Thus, siRNAs and/or shRNAs may become a novel and promising approach to treat skin cancers at an early stage.


Infant acute lymphoblastic leukemia with t(11;16)(q23;p13.3) and lineage switch into acute monoblastic leukemia.
Rearrangements of the mixed-lineage leukemia (MLL) gene have been associated with a poor prognosis in infant acute lymphoblastic leukemia (ALL). Previously, MLL translocations involving the CREP-binding protein (CREBBP) gene at chromosome band 16p13.3 have primarily been reported in treatment-related acute myeloid leukemia, after chemotherapy for other primary malignancies using topoisomerase II inhibitors. We report a case of de novo infant ALL with t(11;16)(q23;p13.3). After chemotherapy, this patient developed an acute monoblastic leukemia (M5b) with retention of the t(11;16)(q23;p13.3), indicating that this is a lineage switch of the original leukemic clone. To our knowledge, these findings have not been previously reported.


Variation in the ovine cortisol response to systemic bacterial endotoxin challenge is predominantly determined by signalling within the hypothalamic-pituitary-adrenal axis.
Bi-directional communication between the neuroendocrine and immune systems is designed, in part, to maintain or restore homeostasis during physiological stress. Exposure to endotoxin during Gram-negative bacterial infection for example, elicits the release of pro-inflammatory cytokines that activate the hypothalamic-pituitary-adrenal axis (HPAA). The secretion of adrenal glucocorticoids subsequently down regulates the host inflammatory response, minimizing potential tissue damage. Sequence and epigenetic variants in genes involved in regulating the neuroendocrine and immune systems are likely to contribute to individual differences in the HPAA response, and this may influence the host anti-inflammatory response to toxin exposure and susceptibility to inflammatory disease. In this study, high (HCR) and low (LCR) cortisol responders were selected from a normal population of 110 female sheep challenged iv with Escherichia coli endotoxin (400 ng/kg) to identify potential determinants that contribute to variation in the cortisol response phenotype. This phenotype was stable over several years in the HCR and LCR animals, and did not appear to be attributed to differences in expression of hepatic immune-related genes or systemic pro-inflammatory cytokine concentrations. Mechanistic studies using corticotrophin-releasing factor (0.5 microg/kg body weight), arginine vasopressin (0.5 microg/kg), and adrenocorticotropic hormone (0.5 microg/kg) administered iv demonstrated that variation in this phenotype is largely determined by signalling within the HPAA. Future studies will use this ovine HCR/LCR model to investigate potential genetic and epigenetic variants that may contribute to variation in cortisol responsiveness to bacterial endotoxin.


Impact of supragingival therapy on subgingival microbial profile in smokers versus non-smokers with severe chronic periodontitis.
The aim of this study was to assess subgingival microbiological changes in smokers versus non-smokers presenting severe chronic periodontitis after supragingival periodontal therapy (ST). Non-smokers (n=10) and smokers (n=10) presenting at least nine teeth with probing pocket depth (PPD) (≥5 mm), bleeding on probing (BoP), and no history of periodontal treatment in the last 6 months were selected. Clinical parameters assessed were plaque index (PI), BoP, PPD, relative gingival margin position (rGMP) and relative clinical attachment level (rCAL). Subgingival biofilm was collected before and 21 days after ST. DNA was extracted and the 16S rRNA gene was amplified with the universal primer pair, 27F and 1492R. Amplified genes were cloned, sequenced, and identified by comparison with known 16S rRNA sequences. Statistical analysis was performed by Student's t and Chi-Square tests (α=5%). Clinically, ST promoted a significant reduction in PI and PPD, and gain of rCAL for both groups, with no significant intergroup difference. Microbiologically, at baseline, data analysis demonstrated that smokers harbored a higher proportion of Porphyromonas endodontalis, Bacteroidetes sp., Fusobacterium sp. and Tannerella forsythia and a lower number of cultivated phylotypes (p<0.05). Furthermore, non-smokers featured significant reductions in key phylotypes associated with periodontitis, whereas smokers presented more modest changes. Within the limits of the present study, ST promoted comparable clinical improvements in smokers and non-smokers with severe chronic periodontitis. However, in smokers, ST only slightly affected the subgingival biofilm biodiversity, as compared with non-smokers.


Nematode burdens of alpacas sharing grazing with sheep in New Zealand.
Sheep and alpacas of similar age groups (6, 18 and 36+ months) were grazed for 16 weeks on pasture contaminated by lambs. Faecal egg counts, bulked larval cultures, lungworm larvae in faeces, dag scores, liveweight changes and nematode larvae on pasture were measured. Chabertia, Oesophagostomum, Cooperia, Ostertagia and Haemonchus and Trichostrongylus larvae were cultured from both the sheep and the alpacas. For the respective age groups, the alpacas had lower liveweight gains (10, 32 and 47 g/d vs 88, 84 and 120 g/d), peak faecal egg counts (384, 50 and 60 epg vs 1500, 500 and 140 epg) and faecal contamination of the perineum than the same ages of sheep. These results suggest alpacas became less affected with gastrointestinal nematodes than sheep.


WITHDRAWN: Allergen specific immunotherapy induced multi-organ failure.
This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.


Chronic low intake of protein or vitamins increases the intestinal epithelial cell apoptosis in Wistar/NIN rats.
Malnutrition decreases antioxidant defense and increases oxidative stress in the intestine. We studied the effects of long-term restriction of food, protein, and vitamins on intestinal epithelial cell (IEC) apoptosis and the underlying mechanisms. Weanling, Wistar/NIN male rats were fed ad libitum with a control diet, 75% protein-restricted diet, or 50% vitamin-restricted diet for 20 wk. The food-restricted group received 50% of the diet consumed by control rats. IEC apoptosis was monitored by morphometry, Annexin V binding, M30 CytoDeath assay, and DNA fragmentation. Structural and functional integrity of the villus were assessed by the ratio of villus height to crypt depth, and alkaline phosphatase and lys, ala-dipeptidyl aminopeptidase activities, respectively. Oxidative stress parameters, caspase-3 activity, and expression of Bcl-2 and Bax were determined to assess the probable mechanisms of altered apoptosis. Protein and vitamin restrictions but not food restriction significantly increased IEC apoptosis and only vitamin restriction altered structural and functional integrity of villi. Increased levels of protein carbonyls, thiobarbituric acid reactive substances, and caspase-3 activity along with decreased glutathione levels and Bcl-2 expression were observed in IECs of these rats, whereas food restriction did not affect these parameters. Protein restriction increased only IEC apoptosis, whereas vitamin restriction also affected the structure and function of villi. Modulation of the pathway mediated by mitochondria through increased oxidative stress appears to be the probable mechanism underlying this effect.


Improved operative exposure of infrapopliteal vessels in combined vascular and orthopedic injuries.
In a series of six patients, a single-bar anterior external fixator was used to treat combined vascular and orthopedic injuries of the leg. This method of external fixation facilitated operative vascular exposure without sacrificing bony stability or alignment. It may be the preferred method of external fixation for these complex injuries.


Dietary exposure of copper and zinc oxides nanoparticles affect the fitness, enzyme activity, and microbial community of the model insect, silkworm Bombyx mori.
Copper and Zinc oxides nanoparticles (CuO and ZnO NPs, respectively) are among the most produced and commonly used engineered nanomaterials. They can be released into the environment, thereby causing health concerns and risks to biodiversity that indicate a need to evaluate their toxicological effects in a complex situation. Here, we used the insect model organism silkworm Bombyx mori to address the concerns about the biological effects associated with dietary exposure of CuO and ZnO NPs. ICP-MS analysis revealed significant accumulation of Cu and Zn (the latter being more accumulated) in silkworms' tissues (gut, fat body, silk gland, and malpighian tubule), and some elimination through feces in the respective NPs-exposed groups. NPs-exposures led to a decrease in larval body mass, survivorship, and cocoon production, where the effects of ZnO NPs were more pronounced. We also found that NPs-exposure induced gene expression changes (Attacin, lysozyme, SOD, and Dronc) and altered the activities of antioxidant enzymes (SOD, GST, and CAT), as well as impaired nutrient metabolism (alpha-amylase). Given their antibacterial property, CuO and ZnO NPs decreased species richness and diversity of the gut bacterial community and shifted their configuration to overt microbiome i.e., decreased abundance of probiotics (e.g., Acetobacter) and increased pathobionts (e.g., Pseudomonas, Bacillus, Escherichia, Enterococcus, Ralstonia, etc.) proportions. Overall, this integrated study revealed the unintended negative effects of CuO and ZnO NPs on silkworms and highlighted the potential to inevitably affect all living things due to intensive and possible mishandling of nanomaterials.


Target-specific, histology-independent, randomized discontinuation study of lapatinib in patients with HER2-amplified solid tumors.
To explore the activity of lapatinib with a novel trial design focused on the drug target rather than on histology. Patients with HER2 amplified gastro-esophageal, bladder, ovarian, or uterine tumors were enrolled into a double-blinded randomized discontinuation study of lapatinib 1,500 mg PO daily. The planned sample size was 250 patients with HER2 amplified tumors, with the goal of randomizing 100 patients with stable disease (SD) at week 12 to either lapatinib or placebo. Patients responding after 12 weeks continued on lapatinib; those who progressed were discontinued from study. The primary objectives were response rate after 12 weeks and the percentage of patients who remained progression free 12 weeks after randomization to placebo versus lapatinib. Secondary objectives were duration of response and determination of the incidence of HER2 amplification in multiple tumor types. A total of 141 patients were screened and 32 patients with HER2 amplified tumors were enrolled. At week 12, 1 (3%) patient had a complete response, 9 (28%) had stable disease, 20 (63%) had progressive disease, and 2 (6%) were unknown. Only 7 patients with SD underwent randomization. The low response rate coupled with slow screening and enrollment led to early study closure. Basing trial eligibility on the presence of a genetic target, versus histologic classification, is challenging. While HER2 amplifications appear to be prevalent in select non-breast tumors, lapatinib monotherapy is associated with modest activity. The target-specific histology-independent randomized discontinuation design still merits consideration for targets clearly implicated in "oncogene addiction".


Regioselective and stereoselective cleavage of cis-4-methyl-2-vinyl-l,3-dioxane by alkyllithiums.
The reaction of alkyllithiums with cis-4-methyl-2-vinyl-1,3-dioxane proceeds in virtually quantitative yield via a formal S(N)2' ring opening that proceeds in a regioselective manner to give preferential cleavage of the C(2)-O(l) bond remote from the 4-methyl substituent, affording the E-enol ether as the major stereoisomer. A two-step mechanism, involving addition of the alkyllithium followed by syn-elimination of lithium alkoxide, is suggested to account for the stereochemical outcome.


The effect of Artemisia annua L. extract on microbiologically influenced corrosion of A36 steel caused by Pseudomonas aeruginosa.
The protective effect of A. annua against microbiologically influenced corrosion (MIC) of A36 steel caused by P. aeruginosa (PA) in a simulated marine environment was investigated using electrochemical, spectroscopic, and surface techniques. PA was found to accelerate the local dissolution of A36 which led to the formation of a porous α-FeOOH and γ-FeOOH surface layer. 2D and 3D profiles of treated coupons, obtained by optical profilometer, revealed the formation of crevices in the presence of PA. On the contrary, adding A. annua to the biotic medium led to the formation of a thinner, more uniform surface without significant damage. Electrochemical data showed that the addition of A. annua prevented the MIC of A36 steel with an inhibition efficiency of 60%. The protective effect was attributed to the formation of a more compact Fe3O4 surface layer, as well as the adsorption of phenolics, such as caffeic acid and its derivatives on the A36 steel surfaces, as detected by FTIR and SEM-EDS analysis. ICP-OES confirmed that Fe and Cr species more readily diffuse from A36 steel surfaces incubated in biotic media (Fe; 1516.35 ± 7.94 μg L-1 cm-2, Cr; 11.77 ± 0.40 μg L-1 cm-2) compared to the inhibited media (Fe; 35.01 ± 0.28 μg L-1 cm-2, Cr; 1.58 ± 0.01 μg L-1 cm-2).


Trauma-registry survival outcome follow up: 30 days is mandatory and appears sufficient.
Thirty-day in-hospital mortality is a common outcome measure in trauma-registry research and benchmarking. However, this does not include deaths after hospital discharge before 30 days or late deaths beyond 30 days since the injury. To evaluate the reliability of this outcome measure, we assessed the timing and causes of death during the first year after major blunt trauma in patients treated at a single tertiary trauma center. We used the Helsinki Trauma Registry to identify severely injured (NISS ≥ 16) blunt trauma patients during 2006 to 2015. The Population Register center of Finland provided the mortality data for patients and Statistics Finland provided the cause of death information from death certificates. Disease, work-related disease, medical treatment, and unknown cause of death were considered as non-trauma related deaths. We divided the 1-year study period into the following three categories: in-hospital death before 30 days (Group 1), death after discharge but within 30 days (Group 2), and death 31 to 365 days since admission (Group 3). We included 3557 patients with a median NISS of 29. Altogether, 21.8% (776/3557) patients died during the first year since the injury. Of these non-survivors, 12.7% (450) were in Group 1, 4.0% (141) in Group 2, and 5.2% (185) in Group 3. Non-traumatic deaths not directly related to the injury increased substantially as the time from the injury increased and were 2.0% (9/450) in Group 1, 13.5% (19/141) in Group 2, and 35.7% (66/185) in Group 3. Thirty-day mortality is a proper outcome that measures survival after severe blunt trauma. However, applying only in-hospital mortality instead of actual 30-day mortality may exclude non-survivors who die at another facility before day 30. This could result in over-optimistic benchmarking results. On the other hand, extending the follow-up period beyond 30 days increases the rate of non-traumatic deaths. By combining data from different registries, it is possible to address this challenge in current trauma-registry research caused by lack of follow up.


Masculinity and food ideals of men who live alone.
Most studies of social aspects of foods have focused on people in multi-person households, often from the perspective of women. Little is known about the food-related experiences of men who live alone. We therefore conducted a qualitative study with 12 men aged 27-47 who lived alone in Vancouver, Canada. Our goals were to explore their food-related ideals, and their perceptions of how those ideals relate to their actual food practices, the context of living alone, and masculine identities. Data were collected through food journals and semi-structured interviews. The men's ideals included being conscious and organized with respect to food so that they could regularly eat meals prepared at home from healthy ingredients. Eating with others was considered an ideal food context, where the meal became an "occasion", as opposed to "just eating". Participants believed that their eating habits often did not live up to these ideals because of lack of time and because the context of living alone was not conducive to eating well. However, they thought their habits were better than what they perceived to be the habits of a stereotypical bachelor who does not know how to cook and has a 'who cares' attitude towards food.


Pirfenidone and nintedanib for pulmonary fibrosis in clinical practice: Tolerability and adverse drug reactions.
The real-world tolerability of pirfenidone and nintedanib in non-clinical trial patients is unknown. Many patients with pulmonary fibrosis have significant medical co-morbidities or baseline characteristics that exclude them from clinical trial participation. We conducted a retrospective chart review study on subjects prescribed nintedanib or pirfenidone for pulmonary fibrosis treatment (any aetiology) from September 2014 to February 2016. A total of 186 subjects were included: 129 received pirfenidone and 57 were prescribed nintedanib and followed up for mean observation periods of 52 ± 17 weeks for pirfenidone and 41 ± 15 weeks for nintedanib. The primary outcome was drug discontinuation as a result of an adverse event. Subjects had significant respiratory impairment at baseline, 63% required home oxygen therapy and mean diffusion capacity of carbon monoxide (DLCO) was 36 ± 14% predicted. Drug discontinuation as a result of an adverse event occurred in 20.9% of subjects on pirfenidone and 26.3% on nintedanib. Drug discontinuation rates for both pirfenidone and nintedanib did not significantly differ from corresponding large clinical trials (ASCEND/CAPACITY and INPULSIS 1 and 2, respectively). Adverse events that occurred with highest frequency on pirfenidone were nausea (26.4%), rash/photosensitivity (14.7%) and dyspepsia/gastroesophageal reflux disease (GERD) (12.4%). Diarrhoea (52.6%) and nausea (29.8%) were reported most often with nintedanib therapy. Patients with pulmonary fibrosis treated with nintedanib or pirfenidone in routine clinical practice had drug tolerability and adverse event profiles comparable with subjects enrolled in clinical trials despite having a greater degree of respiratory impairment and a high prevalence of co-morbid medical conditions.


[Hyperreactive malarial splenomegaly: three clinical cases and literature review].
Hyperreactive malarial splenomegaly (HMS) is the chronic stage of a long-term stimulation of the immune system secondary to plasmodial infections, more frequently in genetically predisposed patients. HMS is a leading cause of large tropical splenomegaly in endemic zones but has been described in immigrants from Africa and in some European expatriates living in endemic countries. Diagnostic criteria include: long-term stay in a endemic zone, often large splenomegaly, high IgM titer, high antiplasmodial antibody titer, regression by at least 40% of splenomegaly six months after curative antimalarial treatment. In tropical settings, B-cell lymphoma and splenic lymphoma are the main differential diagnoses, which may be identified by a clonality analysis. Recent studies suggest that HMS can be treated by a short-term antimalarial therapy as long as the patient resides out of a malarial endemic country.


Stabilities of the C-H...O bonded complexes of the haloforms HCClnF3-n (n = 0-3) with dimethyl ether, oxirane, and acetone: an experimental and theoretical study.
Complexation enthalpies of the complexes of the haloforms HCCl(n)F(3-)(n) (n = 0-3) with dimethyl ether, oxirane, and acetone have been determined in liquid krypton and/or liquid argon using infrared spectroscopy. The same quantities were derived starting from ab initio complexation energies, calculated at the MP2=FULL/aug-cc-VTZ level, and by correcting these energies for thermodynamic and solvent contributions. The two sets of data are compared and discussed.


The time course of hippocampal cholinergic innervation in the developing hypothyroid rat. A combined histochemical and biochemical study of acetylcholinesterase activity.
Development of cholinergic innervation in the hippocampal formation of normal and hypothyroid rats was studied by a combined biochemical and histochemical analysis of acetylcholinesterase (AChE) activity in normal and hypothyroid rats. The normal developmental pattern of cholinergic activity suggests an entrance of septal cholinergic fibers from the fimbria to different zones of the hippocampal formation mainly during the first postnatal week. By 10 days of age, the regional distribution of staining was similar to that in the adult. Thereafter, the intensity of staining increased without major changes in the laminar organization. As shown by closely related histochemical and biochemical findings, hypothyroidism led to a delay in the arrival of cholinergic afferences and a possible subsequent cholinergic hyperinnervation of the hippocampal formation in adulthood. These results are discussed taking into account the time course of structural development in the two synaptic compartments, namely the extrinsic septal neurons and intrinsic pyramidal and granule cells, in comparison with the development of thyroid function.


Ischemic-reperfusion syndromes: biochemical and immunologic rationale for IL-1 targeted therapy.
Ischemic-reperfusion injury (IRI) can affect many organ systems. Examples include strokes, coronary occlusion, accidental hypothermia, compartment syndrome and neonatal hypoxia. To date no mechanism has been fully accepted to explain acute inflammation associated with IRI. There is circumstantial evidence from animal and human ex-vivo cardiac experiments to support the hypothesis that acute inflammation associated with IRI is in part caused by IL-1 beta and/or IL-1 alpha secretion. Danger signal formation, such as uric acid/calcium pyrophosphate crystallization and other cellular stresses, may occur in IRI. These in turn may stimulate innate immune pathways (i.e. cryopyrin-inflammasome; and/ or toll-like receptors 2 and 4) to secrete IL-1 beta. Most IL-1 targeted therapy studies have focused on chronic human diseases and hopefully this discussion will create a framework to encourage use of this therapy in acute inflammation associated with IRI.


Photosynthesis in Chondrus crispus: the contribution of energy spill-over in the regulation of excitonic flux.
Chondrus crispus is a species of red algae that grows on rocks from the middle intertidal into the subtidal zones of the North Atlantic coasts. As such, it has to cope with strongly variable abiotic conditions. Here we studied the response of the photosynthetic apparatus of this red alga to illumination. We found that, as previously described in the case of the unicellular alga Rhodella violacea (E. Delphin et al., Plant Physiol. 118 (1998) 103-113), a single multi-turnover saturating pulse of light is sufficient to induce a strong quenching of fluorescence. To elucidate the mechanisms underlying this fluorescence quenching, we combined room temperature and 77K fluorescence measurements with absorption spectroscopy to monitor the redox state of the different electron carriers in the chain. In addition, we studied the dependence of these various observables upon the excitation wavelength. This led us to identify energy spill-over from Photosystem II to Photosystem I rather than a qE-type non-photochemical quenching as the major source of fluorescence quenching that develops upon a series of 200ms pulses of saturating light results, in line with the conclusion of Ley and Butler (Biochim. Biophys. Acta 592 (1980) 349-363) from their studies of the unicellular red alga Porphyridium cruentum. In addition, we show that the onset of this spill-over is triggered by the reduction of the plastoquinone pool.


Biallelic Mutations in PATL2 Cause Female Infertility Characterized by Oocyte Maturation Arrest.
Oocyte maturation arrest results in female infertility, but the genetic determinants of human oocyte maturation arrest remain largely unknown. Previously, we identified TUBB8 mutations responsible for human oocyte maturation arrest, indicating the important role of genetic factors in the disorder. However, TUBB8 mutations account for only around 30% of individuals with oocyte maturation arrest; thus, the disorder is likely to involve other genetic factors that are as yet unknown. Here, we initially identified a homozygous nonsense mutation of PATL2 (c.784C>T [p.Arg262∗]) in a consanguineous family with a phenotype characterized by human oocyte germinal vesicle (GV) arrest. Subsequent mutation screening of PATL2 in a cohort of 179 individuals identified four additional independent individuals with compound-heterozygous PATL2 mutations with slight phenotypic variability. A genetic burden test further confirmed the genetic contribution of PATL2 to human oocyte maturation arrest. By western blot in HeLa cells, identification of splicing events in affected individuals' granulosa cells, and immunostaining in affected individuals' oocytes, we provide evidence that mutations in PATL2 lead to decreased amounts of protein. These findings suggest an important role for PATL2 mutations in oocyte maturation arrest and expand our understanding of the genetic basis of female infertility.


Divergent trends in suicide mortality by Autonomous Community and sex (1980-2016).
To analyse the changes in mortality trends by suicide according to Autonomous Community and sex in Spain during the period 1980-2016 using joinpoint regression models. Mortality data were obtained from the Instituo Nacional de Estadística. For each Spanish autonomous community and sex, crude and standardised rates were calculated. The joinpoint analysis was used to identify the best-fitting points where a statistically significant change in the trend occurred. The joinpoint analysis allows to differentiate areas in which the rates remain stable in men (Cantabria, Castilla-La Mancha) and women (Canary and Cantabria) throughout the study period and others with a continued decline (Extremadura in both men and women and Castilla-La Mancha in women). In those communities where changes in the trend are observed, in almost all of them, there is a first period of increase in rates in both men and women. The most recent trends show divergences between the different autonomous communities and, in men, Andalusia, the Canary Islands, Castilla-León, the Valencian Community, Galicia, Murcia, the Basque Country and La Rioja show significant downward trends, while Catalonia and Madrid show significant increases (2007-2016: 2.4% and 2010-2016: 18.7% respectively). Something similar is observed in women where Andalusia, Castilla y León, Valencian Community, Galicia, Murcia, País Vasco and La Rioja show downward trends while in the Balearic Islands, Catalonia and Madrid the trend is upward (2001-2016: 5.0%; 2006-2016: 4.2% and 2010-2016: 18.7% respectively). Suicide mortality varies widely among the Spanish autonomous communities, both in terms of mortality level and trends. Little is known about the determinants of observed trends and, therefore, more studies are needed.


Lymph-borne dendritic leucocytes do not recirculate, but enter the lymph node paracortex to become interdigitating cells.
Peripheral lymph contains constitutively major histocompatibility complex (MHC) class II-positive, dendritic leucocytes (DL) with a veiled (VC) or dendritic (DC) shape. On the basis of phenotypic similarities to the interdigitating cells (IDC) of T-dependent areas of lymphoid tissues, they are believed, but not yet proven, to be IDC precursors that develop into mature IDC within the draining lymph nodes. In order to determine the relationship between VC/DC and IDC, highly purified radiolabelled peripheral lymph DL (collected by thoracic duct cannulation of mesenteric lymphadenectomized congenitally athymic, nude rats) were reinjected into the veins or footpads of syngeneic recipients. Later, host blood and lymph were collected and the tissues were biopsied and processed for scintillation counting or for light microscopic autoradiography combined with electron microscopy of neighbouring sections. The intravenously injected VC/DC did not enter lymph nodes or Peyer's patches directly from blood and did not re-enter the lamina propria of the gut, but became localized largely in the liver and the spleen. From the liver some of the cells proceeded via afferent lymphatics to the liver and the spleen. From the liver some of the cells proceeded via afferent lymphatics to the coeliac lymph nodes. Similarly, some of the locally injected cells were carried with peripheral lymph to the draining popliteal lymph nodes, although most were retained in the footpads. In the spleen the VC/DC entered the periarteriolar lymphoid sheaths and in the coeliac and popliteal lymph nodes they became localized in the paracortex, where they acquired the morphology of mature IDC.


Cardiac motion recovery using an incompressible B-solid model.
B-spline based deformable model is commonly used in recovering three-dimensional (3D) cardiac motion from tagged MRI due to its compact description, localized continuity and control flexibility. However, existing approaches usually ignore an important well-known fact that myocardial tissue is incompressible. In this paper, we propose to reconstruct 3D cardiac motion from tagged MRI using an incompressible B-solid model. We demonstrate that cardiac motion recovery can be achieved more with greater accuracy by considering both smoothness and incompressibility of the myocardium. Specifically, our incompressible B-solid model is formulated as a 3D tensor product of B-splines, where each piece of B-spline represents a smooth and divergence-free displacement field of myocardium with respect to radial, longitudinal and circumferential direction, respectively. We further formulate the fitting of the incompressible B-solid model as an optimization problem and solve it with a two-stage algorithm. Finally, the 3D myocardium strains are obtained from the reconstructed incompressible displacement fields and visualized in a comprehensive way. The proposed method is evaluated on both synthetic and in vivo human datasets. Comparisons with state-of-the-art methods are also conducted to validate the proposed method. Experimental results demonstrate that our method has a higher accuracy and more stable volume-preserving ability than previous methods, yielding an average displacement error of 0.21 mm and a Jacobian determinant mean of 1.029.


Pharmacokinetics of topical calcineurin inhibitors in adult atopic dermatitis: a randomized, investigator-blind comparison.
We sought to compare pharmacokinetics of pimecrolimus cream 1% and tacrolimus ointment 0.1% in adults with extensive, moderate to severe atopic dermatitis. Secondary end points included efficacy and safety. Patients received twice-daily treatment for 13 days. Blood concentrations of pimecrolimus and tacrolimus were measured at days 1, 5, and 13. Treatment success was defined as an Investigators' Global Assessment score of 0 (clear) or 1 (almost clear). Tacrolimus was detectable in 36% of blood samples and pimecrolimus was detectable in 12%. In patients with measurable blood drug concentrations, systemic exposure to tacrolimus (mean area under the curve(0-10h) < 9.7 ng.h/mL; n = 7) was higher than to pimecrolimus (mean area under the curve(0-10h) < 2.5 ng.h/mL; n = 2). Whole-body treatment success (day 13) was achieved in 1 of 18 (5.6%) and 2 of 19 (10.5%) patients treated with pimecrolimus and tacrolimus, respectively, and face/neck treatment success in 5 of 18 (27.8%) and 5 of 19 (26.3%) patients, respectively. Patients included in the study were adult patients with severe atopic dermatitis. The results and conclusions drawn from this study population may not be applicable for the majority of patients with atopic dermatitis who have mild to moderate disease. Pimecrolimus appears to be associated with lower systemic drug exposure than tacrolimus.


Reduced dislocation rates and excellent functional outcomes with large-diameter femoral heads.
The purpose of this study was to compare the dislocation rates, functional outcomes, and radiographic results between large- and small-diameter femoral head components. A total of 225 patients (248-hips) who received total hip arthroplasties with large-diameter components (36mm or greater) were compared to 501 patients (559-hips) who received smaller diameter components (less than 36mm). Rates of dislocation and revision, Harris hip score, radiographic findings, and complications were compared between the groups. At a mean follow-up of approximately 5years, a significantly higher rate of dislocation was noted in the small-diameter group (10 of 559) compared to the large-diameter group (0 of 248). The overall implant survivorship, mean Harris hip scores, complication rates, and radiographic outcomes were similar for the two groups. The authors believe that large-diameter components significantly reduced the risk of dislocation in susceptible patients, while preserving good to excellent functional outcomes.


Inducible expression of hyperactive Syk in B cells activates Blimp-1-dependent terminal differentiation.
The non-receptor protein tyrosine kinase Syk (spleen tyrosine kinase) is an important mediator of signal transduction in B cells. By acting downstream of the B-cell antigen receptor, Syk promotes signaling pathways involved in proliferation, differentiation and survival of B cells. To study the oncogenic potential of Syk, we generated a mouse model for the inducible expression of the leukemia-derived TEL-Syk fusion protein exhibiting constitutive kinase activity. To achieve B-cell-specific expression of TEL-Syk in adult mice, we used a tamoxifen-inducible Cre mouse line. This study shows that inducible expression of TEL-Syk in B cells leads to transient proliferation and subsequent plasma cell differentiation. However, it does not lead to B-cell transformation. Instead, Syk activation induces the tumor suppressor B-lymphocyte-induced maturation protein-1 (Blimp-1), which interferes with the expression of the antiapoptotic protein Bcl-2. Combined induction of TEL-Syk with transgenic expression of Bcl-2 results in a severe phenotype and plasma cell expansion. Our results suggest that deregulated Syk activity by itself is not sufficient for the transformation of B cells, as downstream effectors, such as Blimp-1, limit the survival and expansion of the activated B cell.


Magnetic resonance imaging brain yield in developmental delay: A developing country perspective.
Developmental delay (DD) is an important neuro-morbidity in children affecting the quality of life. MRI plays a crucial role by delineating the underlying structural, metabolic, and genetic abnormalities. To determine the yield of MRI brain in delineating the various underlying abnormalities and etiological factors in children with DD and to correlate these findings with the clinical presentation. This cross-sectional study enrolled 50 children with the developmental delay between 6 months to 6 years of age. The mean age was 31.32 ± 20.56 months. The sensitivity of MRI was 72%. 81.3% of the children with microcephaly had abnormal MRI. The most common underlying etiology was hypoxic-ischemic encephalopathy (42%), followed by congenital/developmental defects and metabolic diseases (10% each). The most commonly involved region of the cerebral cortex was the occipital lobe (44%) because of the high occurrence of coexisting hypoglycemic brain injury, which is extremely common in developing countries and rare in developed countries, with 80% of them having visual abnormalities. Frontal lobe involvement was significantly more in children with abnormal motor findings and behavioral changes. Cortical grey matter abnormalities were significantly more in children with seizures. It is to be emphasized that children with developmental delays should be evaluated with MRI whenever possible. Apart from hypoxic-ischemic encephalopathy, other etiologies should also be looked for.


Interhospital Transfer of Critically Ill Patients Because of Coronavirus Disease 19-Related Respiratory Failure.
Interfacility transfer of patients with coronavirus disease 2019-related acute respiratory failure is high risk because of the severity of respiratory failure and potential for crew exposure. This article describes a hospital-based transport team's experience with interfacility transport of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-positive patients. A retrospective study of transports for respiratory failure caused by SARS-CoV-2 was performed. All transports were performed by a single critical care transport team. The team was already trained in advanced mechanical ventilation, blood gas interpretation, and management of shock. Guidance from the Centers for Disease Control and Prevention was followed regarding the use of personal protective equipment. Twenty patients were enrolled. The average patient age was 47 years (standard deviation [SD] = 12 years). The average Acute Physiology and Chronic Health Evaluation and Sequential Organ Failure Assessment scores were 10 (SD = 4) and 24 (SD = 7), respectively. The average transport distance and time were 18 miles (SD = 9 miles) and 25 minutes (SD = 11 minutes), respectively. Nineteen patients were intubated, 9 of whom required advanced ventilation. Two patients were transported prone. One patient experienced unintentional extubation upon transfer from the stretcher to the destination facility bed. The patient was reintubated without event. No crewmembers contracted SARS-CoV-2 infection. Interfacility transfer of severely ill SARS-CoV-2-positive patients is safe and feasible.


Explaining bioenergy: representations of jatropha in Kenya before and after disappointing results.
Proponents of Jatropha curcas portrayed the crop as a 'sustainable biofuel' that was less threatening to food security and forests than other energy crops, creating a reputation that helped jatropha projects to multiply quickly throughout the global South. However, many jatropha initiatives failed to thrive and ultimately collapsed. This paper investigates how actors involved with jatropha in Kenya explained their visions of bioenergy at two points in time. In 2009, when many activities were beginning, I interviewed small-scale farmers, NGO staff, researchers, donors, government officials and members of the private sector about their expectations of jatropha as an energy crop. In late 2013, after jatropha activities in the country had dwindled, I re-interviewed many of the same individuals about their current views and their explanations of the events that had transpired since the initial fieldwork. Synthesizing these two sets of representations provides insight into how biofuel projects have been constructed, negotiated and renegotiated. Early hopes for jatropha rested on the belief that it could achieve many goals simultaneously, but when it failed to meet expectations proponents chose between two strategies: (1) 'unbundling' these goals to pursue separately the various aspirations they had initially attached to jatropha; and (2) seeking a new means of achieving the same bundle of goals. Understanding the choices made by jatropha actors in Kenya contributes to knowledge on the political ecology of biofuels and responsible innovation, and may signal patterns to come as even greater expectations are attached to multi-use feedstocks in pursuit of the bioeconomy.


A quantitative estimate of the maximum amount of light-induced Ca2+ release in Drosophila photoreceptors.
Simultaneous measurements of the light-induced current (LIC) and cytosolic Ca2+ (using INDO-1) were made in Drosophila photoreceptors. In the presence of 1.5 mM Cao2+, the UV light used to measure INDO-1 fluorescence saturated the LIC and induced a large Ca2+ rise. In the absence of extracellular Ca2+ and with Na+ replaced by N-methyl-D-glucamine, the light-induced Ca2+ rise was virtually abolished. A residual rise of about 20 nM is regarded as an upper estimate of Ca2+ released from internal stores. To estimate the Ca2+ flux required to generate such a rise, Ca2+ influx signals in response to weak light steps (500 ms LED stimulus) were measured in the presence of external Ca2+. The relationship between [Ca(in)] and the total charge carried during the LIC had a slope of 2.7 nM pC-1. Assuming that 50% of the LIC is carried by Ca2+ and that the single-channel Ca2+ current carried by the InsP3 receptor is 0.04 pA, it was estimated that about 350 InsP3 receptors should have been sufficient to generate a Ca2+ rise of 20 nM within 500 ms. By contrast, the current activated by the UV measuring light was equivalent to the activation of at least 5000 quantum bumps, making it unlikely that InsP3-induced Ca2+ release could have been the causal event for excitation under these conditions.


Leaf color is fine-tuned on the solar spectra to avoid strand direct solar radiation.
The spectral distributions of light absorption rates by intact leaves are notably different from the incident solar radiation spectra, for reasons that remain elusive. Incident global radiation comprises two main components; direct radiation from the direction of the sun, and diffuse radiation, which is sunlight scattered by molecules, aerosols and clouds. Both irradiance and photon flux density spectra differ between direct and diffuse radiation in their magnitude and profile. However, most research has assumed that the spectra of photosynthetically active radiation (PAR) can be averaged, without considering the radiation classes. We used paired spectroradiometers to sample direct and diffuse solar radiation, and obtained relationships between the PAR spectra and the absorption spectra of photosynthetic pigments and organs. As monomers in solvent, the spectral absorbance of Chl a decreased with the increased spectral irradiance (W m(-2) nm(-1)) of global PAR at noon (R(2) = 0.76), and was suitable to avoid strong spectral irradiance (λmax = 480 nm) rather than absorb photon flux density (μmol m(-2) s(-1) nm(-1)) efficiently. The spectral absorption of photosystems and the intact thallus and leaves decreased linearly with the increased spectral irradiance of direct PAR at noon (I dir-max), where the wavelength was within the 450-650 nm range (R(2) = 0.81). The higher-order structure of photosystems systematically avoided the strong spectral irradiance of I dir-max. However, when whole leaves were considered, leaf anatomical structure and light scattering in leaf tissues made the leaves grey bodies for PAR and enabled high PAR use efficiency. Terrestrial green plants are fine-tuned to spectral dynamics of incident solar radiation and PAR absorption is increased in various structural hierarchies.