NRG1 fusions in breast cancer.
NRG1 gene fusions may be clinically actionable, since cancers carrying the fusion transcripts can be sensitive to tyrosine kinase inhibitors. The NRG1 gene encodes ligands for the HER2(ERBB2)-ERBB3 heterodimeric receptor tyrosine kinase, and the gene fusions are thought to lead to autocrine stimulation of the receptor. The NRG1 fusion expressed in the breast cancer cell line MDA-MB-175 serves as a model example of such fusions, showing the proposed autocrine loop and exceptional drug sensitivity. However, its structure has not been properly characterised, its oncogenic activity has not been fully explained, and there is limited data on such fusions in breast cancer. We analysed genomic rearrangements and transcripts of NRG1 in MDA-MB-175 and a panel of 571 breast cancers. We found that the MDA-MB-175 fusion-originally reported as a DOC4(TENM4)-NRG1 fusion, lacking the cytoplasmic tail of NRG1-is in reality a double fusion, PPP6R3-TENM4-NRG1, producing multiple transcripts, some of which include the cytoplasmic tail. We hypothesise that many NRG1 fusions may be oncogenic not for lacking the cytoplasmic domain but because they do not encode NRG1's nuclear-localised form. The fusion in MDA-MB-175 is the result of a very complex genomic rearrangement, which we partially characterised, that creates additional expressed gene fusions, RSF1-TENM4, TPCN2-RSF1, and MRPL48-GAB2. We searched for NRG1 rearrangements in 571 breast cancers subjected to genome sequencing and transcriptome sequencing and found four cases (0.7%) with fusions, WRN-NRG1, FAM91A1-NRG1, ARHGEF39-NRG1, and ZNF704-NRG1, all splicing into NRG1 at the same exon as in MDA-MB-175. However, the WRN-NRG1 and ARHGEF39-NRG1 fusions were out of frame. We identified rearrangements of NRG1 in many more (8% of) cases that seemed more likely to inactivate than to create activating fusions, or whose outcome could not be predicted because they were complex, or both. This is not surprising because NRG1 can be pro-apoptotic and is inactivated in some breast cancers. Our results highlight the complexity of rearrangements of NRG1 in breast cancers and confirm that some do not activate but inactivate. Careful interpretation of NRG1 rearrangements will therefore be necessary for appropriate patient management.


Expanding targets for a metabolic therapy of cancer: L-asparaginase.
The antitumour enzyme L-asparaginase (L-asparagine amidohydrolase, EC 188.8.131.52, ASNase), which catalyses the deamidation of L-asparagine (Asn) to L-aspartic acid and ammonia, has been used for many years in the treatment of acute lymphoblastic leukaemia. Also NK tumours, subtypes of myeloid leukaemias and T-cell lymphomas respond to ASNase, and ovarian carcinomas and other solid tumours have been proposed as additional targets for ASNase, with a potential role for its glutaminase activity. The increasing attention devoted to the antitumour activity of ASNase prompted us to analyse recent patents specifically concerning this enzyme. Here, we first give an overview of metabolic pathways affected by Asn and Gln depletion and, hence, potential targets of ASNase. We then discuss recent published patents concerning ASNases. In particular, we pay attention to novel ASNases, such as the recently characterised ASNase produced by Helicobacter pylori, and those presenting amino acid substitutions aimed at improving enzymatic activity of the classical Escherichia coli enzyme. We detail modifications, such as natural glycosylation or synthetic conjugation with other molecules, for therapeutic purposes. Finally, we analyse patents concerning biotechnological protocols and strategies applied to production of ASNase as well as to its administration and delivery in organisms.


Impaired local Vitamin D3 metabolism contributes to IL-36g overproduction in epithelial cells in chronic rhinosinusitis with nasal polyps.
Vitamin D (VD) possesses immunomodulatory properties, but its role in chronic rhinosinusitis with nasal polyps (CRSwNP) remains poorly studied. Herein, we aim to explore the regulation and function of VD3 in CRSwNP. 25-hydroxyvitamin D3 (25VD3) levels in serum and tissue lysates were detected by ELISA. The expression of VD receptor (VDR) and cytochrome P450 family 27 subfamily B member 1 (CYP27B1), the enzyme that converts 25VD3 to the active 1,25-hydroxyvitamin D3 (1,25VD3), and their expression regulation in human nasal epithelial cells (HNECs) were studied by RT-PCR, western blotting, immunofluorescence, and flow cytometry. RNA sequencing was performed to identify genes regulated by 1,25VD3 in HNECs. HNECs and polyp tissue explants were treated with 1,25VD3, 25VD3, and dexamethasone. 25VD3 levels in serum and nasal tissue lysates were decreased in patients with eosinophilic and noneosinophilic CRSwNP than control subjects. The expression of VDR and CYP27B1 were reduced in eosinophilic and noneosinophilic CRSwNP, particularly in nasal epithelial cells. VDR and CYP27B1 expression in HNECs were downregulated by interferon γ and poly (I:C). Polyp-derived epithelial cells demonstrated an impaired ability to convert 25VD3 to 1,25VD3 than control tissues. 1,25VD3 and 25VD3 suppressed IL-36γ production in HNECs and polyp tissues, and the effect of 25VD3 was abolished by siCYP27B1 treatment. Tissue 25VD3 levels negatively correlated with IL-36γ expression and neutrophilic inflammation in CRSwNP. Reduced systemic 25VD3 level, local 1,25VD3 generation and VDR expression result in impaired VD3 signaling activation in nasal epithelial cells, thereby exaggerating IL-36γ production and neutrophilic inflammation in CRSwNP.


CT image segmentation methods for bone used in medical additive manufacturing.
The accuracy of additive manufactured medical constructs is limited by errors introduced during image segmentation. The aim of this study was to review the existing literature on different image segmentation methods used in medical additive manufacturing. Thirty-two publications that reported on the accuracy of bone segmentation based on computed tomography images were identified using PubMed, ScienceDirect, Scopus, and Google Scholar. The advantages and disadvantages of the different segmentation methods used in these studies were evaluated and reported accuracies were compared. The spread between the reported accuracies was large (0.04 mm - 1.9 mm). Global thresholding was the most commonly used segmentation method with accuracies under 0.6 mm. The disadvantage of this method is the extensive manual post-processing required. Advanced thresholding methods could improve the accuracy to under 0.38 mm. However, such methods are currently not included in commercial software packages. Statistical shape model methods resulted in accuracies from 0.25 mm to 1.9 mm but are only suitable for anatomical structures with moderate anatomical variations. Thresholding remains the most widely used segmentation method in medical additive manufacturing. To improve the accuracy and reduce the costs of patient-specific additive manufactured constructs, more advanced segmentation methods are required.


Functional characterization and novel rickettsiostatic effects of a Kunitz-type serine protease inhibitor from the tick Dermacentor variabilis.
Here we report the novel bacteriostatic function of a five-domain Kunitz-type serine protease inhibitor (KPI) from the tick Dermacentor variabilis. As ticks feed, they release anticoagulants, anti-inflammatory and immunosuppressive molecules that mediate the formation of the feeding lesion on the mammalian host. A number of KPIs have been isolated and characterized from tick salivary gland extracts. Interestingly, we observe little D. variabilis KPI gene expression in the salivary gland and abundant expression in the midgut. However, our demonstration of D. variabilis KPI's anticoagulant properties indicates that D. variabilis KPI may be important for blood meal digestion in the midgut. In addition to facilitating long-term attachment and blood meal acquisition, gene expression studies of Drosophila, legumes, and ticks suggest that KPIs play some role in the response to microbial infection. Similarly, in this study, we show that challenge of D. variabilis with the spotted fever group rickettsia, Rickettsia montanensis, results in sustained D. variabilis KPI gene expression in the midgut. Furthermore, our in vitro studies show that D. variabilis KPI limits rickettsial colonization of L929 cells (mouse fibroblasts), implicating D. variabilis KPI as a bacteriostatic protein, a property that may be related to D. variabilis KPI's trypsin inhibitory capability. This work suggests that anticoagulants play some role in the midgut during feeding and that D. variabilis KPI may be involved as part of the tick's defense response to rickettsiae.


A density functional theory study of the hydrogenation and reduction of the thio-spinel Fe3S4{111} surface.
The mineral greigite, Fe3S4, shows promising electro-reduction activity, especially towards carbon dioxide conversion to small organic molecules. We have employed density functional theory calculations with correction for the long-range dispersion forces to investigate the behavior of hydrogen on the greigite{111} surface. We have studied the adsorption, diffusion, surface reduction and associative (i.e. Volmer-Tafel mechanism) and molecular desorption of hydrogen as a function of its coverage. We found that (i) the H ad-atoms adsorb on S sites far from metallic centres in the topmost surface layer; (ii) the reduction of greigite by hydrogen is energetically unfavorable at any surface coverage; and (iii) molecular hydrogen evolution has a transition state at ∼0.5 eV above the energy of the reactants on Fe3S4{111}, which is very similar to the barrier found experimentally on Pt{111}. We have also determined the electrode potential under room conditions at which the H2 evolution reaction becomes energetically barrierless.


[Insulin, renin-angiotensin system, aldosterone and endothelial dysfunction].
Beyond its metabolic effects, insulin has several actions on the vasculature. Under normal conditions, insulin maintains normal endothelial function, but in the presence of insulin resistance, insulin leads to endothelial dysfunction. Insulin releases nitric oxide, which promotes an antiatherosclerotic, antiinflamatory and vasodilated state. However, in presence of high levels of angiotensin II, insulin activates pathways that lead to atherosclerosis, vasoconstriction and inflammation. We will review the actions of insulin on the vascular system, and its interactions with other vasoactive mediators, such as angiotensin II and endothelin-1.


Effects of carbon dioxide on neurotoxin gene expression in nonproteolytic Clostridium botulinum Type E.
Carbon dioxide is an antimicrobial gas commonly used in modified atmosphere packaging. In the present study, the effects of carbon dioxide on the growth of and neurotoxin production by nonproteolytic Clostridium botulinum type E were studied during the growth cycle. Quantitative reverse transcription-PCR and an enzyme-linked immunosorbent assay were used to quantify expression of the type E botulinum neurotoxin gene (cntE) and the formation of type E neurotoxin. The expression levels of cntE were similar in two strains, with relative expression peaking in the transition between exponential phase and stationary phase. In stationary phase, cntE mRNA expression declined rapidly. The cntE mRNA half-life was calculated to be approximately 9 minutes. Neurotoxin formation occurred in late exponential phase and stationary phase. High carbon dioxide concentrations delayed growth by increasing the lag time and decreasing the maximum growth rate. The effects of carbon dioxide concentration on relative neurotoxin gene expression and neurotoxin formation were significant. Expression of cntE mRNA and the formation of extracellular neurotoxin were twofold higher with a headspace carbon dioxide concentration of 70% (vol/vol) compared to 10% (vol/vol). This finding sheds a new, cautionary light on the potential risks of botulism associated with the use of modified atmosphere packaging.


[Evaluation of thymopoiesis: clinical applications].
In the precedent article, we have described how T-cell generation in the thymus (thymopoiesis) may be currently evaluated through quantification by PCR of T-cell receptor excision circles (TREC) generated by intrathymic random recombination of the gene segments coding for variable parts of T-cell receptor for antigen (TCR). In hematology, TREC methodology helps in a better understanding of immune reconstitution after graft of hematopoietic stem cells: first there is a proliferation of mature T cells present in the graft, then a differentiation of naive T cells. In geriatrics, the homeostasis of the peripheral T-cell repertoire is maintained through proliferation of peripheral memory T cells rather than through thymic generation of naive T cells. In addition, TREC quantification constitutes a novel major tool for deciphering the tight control of thymopoiesis by the neuroendocrine system.


An injectable, thermosensitive and multicompartment hydrogel for simultaneous encapsulation and independent release of a drug cocktail as an effective combination therapy platform.
Combination chemotherapy is potent to combat diseases. Simultaneous and segregated delivery of multiple drugs in a single vehicle is essential to achieve this objective. In the present study, an injectable, thermosensitive and multicompartment hydrogel (MCH) was developed by the facile cooperative and incompatible assembly of PEGylated hydrocarbon nanoparticles with PEGylated fluorocarbon nanoparticles. The cooperative assembly behavior was investigated by fluorescence resonance energy transfer (FRET) technology, and the result demonstrated that the incompatible nanoparticle cores possibly accounted for the multicompartment formation in hydrogel. Paclitaxel and doxorubicin could be easily and separately integrated into the different compartments of MCH serving as a sustained drug cocktail formulation. In vitro drug release indicated drugs were liberated in a simultaneous but independent manner without any effect on each other. In vitro and in vivo antitumor activity indicated that peritumoral injection of drug cocktail encapsulated MCH formulation could well achieve the combination effect, which significantly improved the tumor growth inhibition efficiency as well as minimized the drug-associated side effects compared to intravenous injection of free drug cocktail. Furthermore, such a delivery device would allow precise adjustment of drug dosage to the desired effect, achieve spatial-temporal simultaneous and synchronized presence of combination drugs in the target tissue and obviate repeated drug administrations to improve patient compliance. The thermosensitive multicompartment hydrogel cocktail formulation holds great promise for simultaneous and segregated delivery of multiple bioactive agents for sustained combination therapy.


Antioxidant activity in extracts of 27 indigenous Taiwanese vegetables.
The objectives of this study were to identify the antioxidants and antioxidant axtivity in 27 of Taiwan's indigenous vegetables. Lycium chinense (Lc), Lactuca indica (Li), and Perilla ocymoides (Po) contained abundant quercetin (Que), while Artemisia lactiflora (Al) and Gynura bicolor (Gb) were rich in morin and kaempferol, respectively. Additionally, Nymphoides cristata (Nc) and Sechium edule (Se)-yellow had significantly higher levels of myricetin (Myr) than other tested samples. Cyanidin (Cyan) and malvidin (Mal) were abundant in Gb, Abelmoschus esculentus Moench (Abe), Po, Anisogonium esculentum (Retz.) Presl (Ane), Ipomoea batatas (Ib)-purple, and Hemerocallis fulva (Hf)-bright orange. Relatively high levels of Trolox equivalent antioxidant capacity (TEAC), oxygen radical absorption capacity (ORAC), and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenger were generated from extracts of Toona sinensis (Ts) and Po. Significant and positive correlations between antioxidant activity and polyphenols, anthocyanidins, Que, Myr, and morin were observed, indicating that these phytochemicals were some of the main components responsible for the antioxidant activity of tested plants. The much higher antioxidant activity of Po, Ts, and Ib (purple leaf) may be related to their higher Cyan, Que, and polyphenol content.


Use of preoperative hypnosis to reduce postoperative pain and anesthesia-related side effects.
The purpose of this pilot project was to test the feasibility of hypnosis as a preoperative intervention. The unique features of this study were: (a) use of a standardized nurse-delivered hypnosis protocol, (b) intervention administration immediately prior to surgery in the preoperative holding area, and (c) provision of hypnosis to breast cancer surgery patients receiving general anesthesia. A mixed-method design was used. Data collected from the intervention group and historical control group included demographics, symptom assessments, medication administration, and surgical, anesthesia, and recovery minutes. A semi-structured interview was conducted with the intervention group. A reduction in anxiety, worry, nervousness, sadness, irritability, and distress was found from baseline to postintervention while pain and nausea increased. The results support further exploration of the use of nurse-led preoperative hypnosis.


Reduction in the chondrocyte response to insulin-like growth factor 1 in aging and osteoarthritis: studies in a non-human primate model of naturally occurring disease.
Although the development of osteoarthritis (OA) is closely associated with aging, the mechanism for this association is not clear. This study was designed to determine the effects of aging and OA on the chondrocyte response to stimulation with insulin-like growth factor 1 (IGF-1) in a non-human primate model of naturally occurring OA. Chondrocytes were isolated from cartilage removed separately from the medial and lateral femoral condyles and tibial plateaus of cynomolgus monkeys at the time of necropsy. Each joint site was scored histologically on a scale of 0-7 for OA pathologic changes. Isolated chondrocytes were cultured in alginate in serum-free medium and stimulated with IGF-1 or des(1-3) IGF-1, which has a much lower affinity for IGF binding proteins (IGFBP) than IGF-1. Response was measured as the ability to stimulate sulfate and proline incorporation. Cartilage samples from 34 monkeys ranging in age from 6.7 years to 27 years and with histologic scores ranging from 0 to 7 were analyzed. A significant decline in the response to IGF-1 was noted with both increasing age and increasing OA score. Controlling for the OA score, the estimated effect of age on IGF-1 response, measured by total sulfate incorporation, was a decline of 3.81% per year (P = 0.0001), or a 75% decline over 20 years as a monkey ages from young to older adult. Controlling for age, the effect of OA score was significant only for proline incorporation in the alginate matrix (estimated slope coefficient +/-standard error -15.9 +/- 7.2; P = 0.03), suggesting a negative effect of OA on retention of 3H-proline-labeled proteins in the matrix. There was a significantly reduced response to des(1-3) IGF-1 with increasing age, but no effect of OA score on response to des(1-3) IGF-1. There was no effect of age on cell viability. These results demonstrate a significant age-related decline in the chondrocyte response to IGF-1. The finding that increasing OA score was associated with a reduced response to intact IGF-1 but not des(1-3) IGF-1 suggests a role of increased production of inhibitory IGFBP in OA. Since the cells from older animals had a reduced response to both forms of IGF-1, the mechanism of the reduced response with age cannot be attributed to changes in IGFBP. Age-related changes in IGF receptors or, more likely, age-related alterations in intracellular signal transduction may also be involved.


Capillary washboarding during slow drainage of a frictional fluid.
Numerous natural and industrial processes involve the mixed displacement of liquids, gases and granular materials through confining structures. However, understanding such three-phase flows remains a formidable challenge, despite their tremendous economic and environmental impact. To unveil the complex interplay of capillary and granular stresses in such flows, we consider here a model configuration where a frictional fluid (an immersed sedimented granular layer) is slowly drained out of a horizontal capillary. Analyzing how liquid/air menisci displace particles from such granular beds, we reveal various drainage patterns, notably the periodic formation of dunes, analogous to road washboard instability. Considering the competitive role of friction and capillarity, a 2D theoretical approach supported by numerical simulations of a meniscus bulldozing a front of particles provides quantitative criteria for the emergence of those dunes. A key element is the strong increase of the frictional forces, as the bulldozed particles accumulate and bend the meniscus horizontally. Interestingly, this frictional enhancement with the attack angle is also crucial in small-legged animals' locomotion over granular media.


Persistent CSF Rhinorrhoea, Pneumocephalus, and Recurrent Meningitis Following Misdiagnosis of Olfactory Neuroblastoma.
A 41-year-old female patient was admitted with streptococcal meningitis on a background of 5-month history of CSF rhinorrhoea. Imaging revealed an extensive skull base lesion involving the sphenoid and ethmoid sinuses, the pituitary fossa with suprasellar extension and bony destruction. Histological examination of an endonasal transethmoidal biopsy suggested a diagnosis of olfactory neuroblastoma. A profuse CSF leak occurred and the patient developed coliform meningitis. A second endonasal endoscopic biopsy was undertaken which demonstrated the tumour to be a prolactinoma. Following endonasal repair of the CSF leak and lumbar drainage, she developed profound pneumocephalus. The patient underwent three further unsuccessful CSF leak repairs. Definitive control of the CSF leak was finally achieved through a transcranial approach with prolonged lumbar drainage. This case illustrates some of the potentially devastating complications which can occur as a consequence of complex skull base lesions. A multidisciplinary approach may be required to successfully manage such cases.


[Effect of bacterial physiological state and heavy metals on the hyperactive reaction of tobacco leaf].
It has been shown that the initiation and proceeding of hypersensitive reaction on the tobacco leaf under the bacterial effect are influenced by the infectious loading of the pathogen, physiological state of bacteria, their viability, temperature of the environment and heavy metal salts. It has been established that the hypersensitive reaction is most distinctly manifested when 1-3 days bacterium cultures are used. Their activity decreases essentially with age, and 10-14 days old cultures are not able to evoke the hypersensitive reaction. The bacterial cells being inactivated with temperature and UV-irradiation lose the ability to evoke the hypersensitive reaction. The reaction of hypersensitive is inhibited by salts of heavy metals in concentrations 0.1-1% and by high temperature of the environment (30 degrees C).


Effect of ethanol treatment on mechanical properties of heat-polymerized polymethyl methacrylate denture base polymer.
This laboratory study was assessing the nano-mechanical properties (NMP), surface roughness (Sa), and topographic changes caused by ethanol on the surface of heat-polymerized denture base polymers at different time past dough stage. Specimens of heat polymerizing acrylic resin (Interacryl Hot, Interdent, Celje, Slovenia) of size 10×10×3 mm were prepared, wet ground, and polished for uniform smoothness and treated with ethanol in concentrations of 40, 70, and 99.9% for 30, 60, and 120 s and statistical analysis was done. Some statistical significance for Sa were highest with 120 s exposure to 40% ethanol. NMP were the highest for specimens treated with 99.9% ethanol concentration for 120 s, on specimens prepared 30 min past the dough stage. This study suggested that heat-polymerized denture base polymers are prone for changes by ethanol which alters mechanical properties and surface topography. Dough time influenced the ethanol resistance.


OSA Wellness Scale (OWS): A New Health-Related Quality of Life Test in Obstructive Sleep Apnea Patients Treated with Mandibular Advancement Device.
To present a new short self-test, called the OSA wellness scale (OWS), for assessing the health-related quality of life (HRQoL) changes in obstructive apnea syndrome (OSA) patients treated with mandibular advancement device (MAD). 51 OSA patients (8 women and 43 men, mean age 52.3) treated with a fully customizable MAD device (Protrusor) were retrospectively enrolled. Each patient received a home sleep apnea testing (HSAT) at baseline (T0) and after three months of MAD treatment (T1). Two self-test evaluations, the Epworth sleepiness scale (ESS), and OWS were also submitted at T0 and T1. The OWS was a short self-test of 8 questions for evaluating the daytime HRQoL. Patients gave an assessment from 0 to 3 for each question. At the end of the questionnaire, the patients had a score from 0 to 24, resulting from the sum of all 8 scores. The higher the score, the greater the patient's perceived state of discomfort. At T1, a significant decrease in the oxygen desaturation index (ODI) and apnea-hypopnea index (AHI) was shown (p < 0.0001), while no significant changes in body mass index (BMI) were found. Both the ESS and the OWS records showed a significant reduction in daytime sleepiness and HRQoL (p < 0.0001). The OWS could be a useful method to verify and numerically compare the perceived quality of life in OSA patients, before and after MAD therapy.


TOR2A Variants in Blepharospasm.
Genetic factors have been implicated in the pathogenesis of blepharospasm (BSP), a dystonia characterized by excessive blinking and involuntary eyelid closure. Previous research identified a co-segregating deleterious TOR2A variant (GRCh38/hg38, NC_000009.12: g.127733410G>A, NM_001085347.3:c.568C>T, p. Arg190Cys) in three subjects with BSP and three carriers within a multi-generation pedigree. Other TOR2A variants have been reported in patients with dystonia. Sanger sequencing was used to screen a cohort of 307 subjects with isolated BSP or BSP-plus dystonia affecting additional anatomical segments (BSP+). We also utilized computational tools to uniformly assess the deleteriousness and potential pathogenicity of previously reported TOR2A variants. There were no highly deleterious TOR2A variants in the coding or contiguous splice site regions of TOR2A within our cohort of 307 subjects. Highly deleterious variants in TOR2A are rare in patients with BSP/BSP+ phenotypes. Over 300 patients with BSP were screened for variants in TOR2A, a TOR1A (DYT1) homologue. No highly deleterious variants were identified in our cohort. The role of TOR2A in BSP and other forms of dystonia remains indeterminant.


Bone metastasis in a case of primary acquired melanosis with atypia resulting from conjunctiva melanoma.
A 66-year-old woman presented to a tertiary referral centre with a diagnosis of limbal stem cell deficiency secondary to multiple treatments for primary acquired melanosis (PAM) by her referring ophthalmologists. She was known with primary acquired melanosis with severe atypia of the right eye which had been treated with several excisions, cryotherapy and topical chemotherapy. She had PAM over 360° of alternating intensity. Throughout the follow-up period, she developed a cataract and subsequent a retinal detachment for which she underwent a phacovitrectomy on the right eye. There was no nodule present nor did the routine lymph nodes inspection show any enlargement. No abnormalities were observed on intranasal examination. Seven years after the last excision, a metastasis to the spine (Th3) was diagnosed. A complete systemic work-up and a pathological examination did not show any alternative primary melanoma in the body. The original histological material was reviewed again by an ophthalmopathologist. After a new round of stains and immunohistochemistry, the presence of atypical melanocytes in both the epithelium and stroma were found in a scar region leading to a renewed conclusion that the PAM in question was actually the primary melanoma.


Variability in Radiation Dose From Repeat Identical CT Examinations: Longitudinal Analysis of 2851 Patients Undergoing 12,635 Thoracoabdominal CT Scans in an Academic Health System.
The purpose of this study was to conduct longitudinal analyses of radiation dose data from adult patients undergoing clinically indicated, repeat identical thoracoabdominal CT examinations. Radiation dose data were electronically collected from 2851 subjects undergoing 12,635 repeat identical CT scans (mean number of scans per patient, 4.8; range, 2-33) in one health system. Included CT protocols were chest-abdomen-pelvis with contrast administration (n = 4621 CT studies of 1064 patients), abdomen-pelvis with contrast administration (n = 876 CT studies of 261 patients), renal stone (n = 1053 CT studies of 380 patients), and chest (n = 6085 CT studies of 1146 patients) without contrast administration. A radiation-tracking software infrastructure was adopted to extract data from DICOM headers in PACS. Size-specific dose estimate (SSDE) was calculated. A trend was observed toward global reduction in SSDE values with all protocols investigated (chest-abdomen-pelvis slope, -1.78; abdomen-pelvis slope, -0.82; renal stone slope, -0.83; chest slope, -0.47; p < 0.001 for all comparisons). The intraindividual analyses of radiation dose distribution showed widespread variability in SSDE values across the four protocols investigated (chest-abdomen-pelvis mean coefficient of variance, 14.02 mGy; abdomen-pelvis mean coefficient of variance, 10.26 mGy; renal stone mean coefficient of variance, 34.18 mGy; chest mean coefficient of variance, 6.74 mGy). Although there is a trend toward global reduction in radiation doses, this study showed widespread variability in the radiation dose that each patient undergoing identical repeat thoracoabdominal CT protocols absorbs. These data may provide a foundation for the future development of best-practice guidelines for patient-specific radiation dose monitoring.


A new potentiometric ammonium electrode for biosensor construction.
New ammonium-selective membrane electrode based on poly(vinyl chloride) (PVC) membrane containing palmitic acid (a long-chain fatty acid) and nonactin as an ammonium ionophore for the determination of ammonium ions in the 10(-7) - 10(-1) mol/L concentration range was prepared and compared to those of the electrode prepared by using carboxylated PVC. Sebacate was used as a plasticiser for both of the ammonium sensor membranes. The analytical characteristics of the ammonium electrodes was investigated. The effect of pH, buffer concentration, temperature and stirring rate on the response to ammonium electrode was investigated. The linear working range and sensitivity of the electrodes were also determined. Ammonium electrodes give Nernstian response (52-58 mV/p[NH4+]) throughout the ammonium ion concentration range of 10(-1) to 10(-6)M with detection limits of 10(-6)M ammonium ions. The ammonium-selective electrodes prepared by using the PVC membranes containing palmitic acid showed more effective performance than those of the carboxylated PVC. The ammonium ion sensor has potential application in the analysis of ammonium ions for biosensor construction.


Assessing predictors of delayed antenatal care visits in Rwanda: a secondary analysis of Rwanda demographic and health survey 2010.
Early initiation of antenatal care (ANC) can reduce common maternal complications and maternal and perinatal mortality. Though Rwanda demonstrated a remarkable decline in maternal mortality and 98% of Rwandan women receive antenatal care from a skilled provider, only 38% of women have an ANC visit in their first three months of pregnancy. This study assessed factors associated with delayed ANC in Rwanda. This is a cross-sectional study using data collected during the 2010 Rwanda DHS from 6,325 women age 15-49 that had at least one birth in the last five years. Factors associated with delayed ANC were identified using a multivariable logistic regression model using manual backward stepwise regression. Analysis was conducted in Stata v12 applying survey commands to account for the complex sample design. Several factors were significantly associated with delayed ANC including having many children (4-6 children, OR = 1.42, 95% CI: 1.22, 1.65; or more than six children, OR = 1.57, 95% CI: 1.24, 1.99); feeling that distance to health facility is a problem (OR = 1.20, 95% CI: 1.04, 1.38); and unwanted pregnancy (OR = 1.41, 95% CI: 1.26, 1.58). The following were protective against delayed ANC: having an ANC at a private hospital or clinic (OR = 0.29, 95% CI: 0.15, 0.56); being married (OR = 0.85, 95% CI: 0.75, 0.96), and having public mutuelle health insurance (OR = 0.81, 95% CI: 0.71, 0.92) or another type of insurance (OR = 0.33, 95% CI: 0.23, 0.46). This analysis revealed potential barriers to ANC service utilization. Distance to health facility remains a major constraint which suggests a great need of infrastructure and decentralization of maternal ANC to health posts and dispensaries. Interventions such as universal health insurance coverage, family planning, and community maternal health system are underway and could be part of effective strategies to address delays in ANC.


Histone deacetylase 3 depletion in osteo/chondroprogenitor cells decreases bone density and increases marrow fat.
Histone deacetylase (Hdac)3 is a nuclear enzyme that contributes to epigenetic programming and is required for embryonic development. To determine the role of Hdac3 in bone formation, we crossed mice harboring loxP sites around exon 7 of Hdac3 with mice expressing Cre recombinase under the control of the osterix promoter. The resulting Hdac3 conditional knockout (CKO) mice were runted and had severe deficits in intramembranous and endochondral bone formation. Calvarial bones were significantly thinner and trabecular bone volume in the distal femur was decreased 75% in the Hdac3 CKO mice due to a substantial reduction in trabecular number. Hdac3-CKO mice had fewer osteoblasts and more bone marrow adipocytes as a proportion of tissue area than their wildtype or heterozygous littermates. Bone formation rates were depressed in both the cortical and trabecular regions of Hdac3 CKO femurs. Microarray analyses revealed that numerous developmental signaling pathways were affected by Hdac3-deficiency. Thus, Hdac3 depletion in osterix-expressing progenitor cells interferes with bone formation and promotes bone marrow adipocyte differentiation. These results demonstrate that Hdac3 inhibition is detrimental to skeletal health.


Quantifying and Localizing the Mitochondrial Proteome Across Five Tissues in A Mouse Population.
We have used SWATH mass spectrometry to quantify 3648 proteins across 76 proteomes collected from genetically diverse BXD mouse strains in two fractions (mitochondria and total cell) from five tissues: liver, quadriceps, heart, brain, and brown adipose (BAT). Across tissues, expression covariation between genes' proteins and transcripts-measured in the same individuals-broadly aligned. Covariation was however far stronger in certain subsets than others: only 8% of transcripts in the lowest expression and variance quintile covaried with their protein, in contrast to 65% of transcripts in the highest quintiles. Key functional differences among the 3648 genes were also observed across tissues, with electron transport chain (ETC) genes particularly investigated. ETC complex proteins covary and form strong gene networks according to tissue, but their equivalent transcripts do not. Certain physiological consequences, such as the depletion of ATP synthase in BAT, are thus obscured in transcript data. Lastly, we compared the quantitative proteomic measurements between the total cell and mitochondrial fractions for the five tissues. The resulting enrichment score highlighted several hundred proteins which were strongly enriched in mitochondria, which included several dozen proteins were not reported in literature to be mitochondrially localized. Four of these candidates were selected for biochemical validation, where we found MTAP, SOAT2, and IMPDH2 to be localized inside the mitochondria, whereas ABCC6 was in the mitochondria-associated membrane. These findings demonstrate the synergies of a multi-omics approach to study complex metabolic processes, and this provides a resource for further discovery and analysis of proteoforms, modified proteins, and protein localization.


Identifying the critical time points for mental health of asylum seekers and refugees in high-income countries.
High heterogeneity was found in the prevalence rates of mental disorders in adult asylum seekers and refugees in high-income countries. This may be related to different problems. Among them, there is a changing exposure to risk and protective factors for mental health at different phases of these people's life before migration, and during the migratory journey and resettlement. This study aimed at identifying and distinguishing time points in which distinct risk and protective factors for the mental health of asylum seekers and refugees may occur. Systematic review and narrative synthesis. A systematic search was carried out for the period January 2017-August 2019, given the existence of systematic reviews of the evidence up to January 2017. Two hundred and fifty-two studies were identified with our search and 31 studies were included. The critical time points identified are: (a) before the travel; (b) during the travel; (c) at initial settlement in the host country; (d) when attempting to integrate in the host country; (e) when the immigration status is challenged or revoked. Some factors such as sense of belonging in the host country can be risk factors or protective factors depending on the time point. These five critical time points can guide the development and selection of well-timed preventive and treatment interventions. They could also be used to stratify samples in epidemiological studies and meta-analyses. At present, we know much more on risk factors than on protective factors. Knowing more about protective factors may inform the development of interventions to foster them.


Dermatology residency program characteristics that correlate with graduates selecting an academic dermatology career.
To examine the characteristics of 107 dermatology residency programs to determine which factors are correlated with producing academic dermatologists to help reverse the trend of a growing shortage of academic dermatologists. We collected data ranging from total publications to grant funding. Extensive Internet searches were completed to obtain most of the data. Individual programs were contacted as needed to obtain any missing data that were not found on the program's Web site. Dermatology residency programs (departments and divisions) in the United States. Factors that correlated with producing full-time academic dermatologists. We tabulated and analyzed characteristics of 107 dermatology residency programs. Total full-time faculty members in 2004, total publications in 2004, and total publications from January 1, 2001, to December 31, 2004, were the 3 factors most strongly correlated with producing full-time faculty. National Institutes of Health and Dermatology Foundation grants and American Skin Association grant recipients were the 3 characteristics most strongly inversely correlated with producing full-time faculty. Those who entered academic dermatology tended to stay at the same program where they completed a dermatology residency, especially if this program was considered a "smaller" program. The programs' characteristics of total publications in 2004 and from 2001 to 2004 were 2 of the 3 factors most strongly positively correlated with dermatology residents entering academic dermatology. Encouraging residents to publish may be a window to motivate them toward a career in academic dermatology.


Effect of Sodium and Chloride Binding on a Lecithin Bilayer. A Molecular Dynamics Study.
The effect of ion binding on the structural, mechanical, dynamic and electrostatic properties of a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer in a 0.5 M aqueous NaCl solution is investigated using classical atomistic molecular dynamics simulation with different force-field descriptions for ion-ion and ion-lipid interactions. Most importantly, the repulsive Lennard-Jones parameters for the latter were modified, such that approximately similar binding of cations and anions to the lipid membrane is achieved. This was done to qualitatively improve the apparent ion-lipid binding constants obtained from simulations with the original force field (Berger lipids and GROMOS87 ions in combination with the SPC water model) in comparison to experimental data. Furthermore, various parameters characterizing membrane structure, elasticity, order and dynamics are analyzed. It is found that ion binding as observed in simulations involving the modified in comparison to the original force-field description leads to: (i) a smaller salt-induced change in the area per lipid, which is in closer agreement with the experiment; (ii) a decrease in the area compressibility and bilayer thickness to values comparable to a bilayer in pure water; (iii) lipid deuterium order parameters and lipid diffusion coefficients on nanosecond timescales that are very similar to the values for a membrane in pure water. In general, salt effects on the structural properties of a POPC bilayer in an aqueous sodium-chloride solution appear to be reproduced reasonably well by the new force-field description. An analysis of membrane-membrane disjoining pressure suggests that the smaller salt-induced change in area per lipid induced by the new force-field description is not due to the alteration of membrane-associated net charge, but must rather be understood as a consequence of ion-specific effects on the arrangement of lipid molecules.


Airway contraction and cytokine release in isolated rat lungs induced by wear particles from the road and tire interface and road vehicle brakes.
The dominant road traffic particle sources are wear particles from the road and tire interface, and from vehicle brake pads. The aim of this work was to investigate the effect of road and brake wear particles on pulmonary function and biomarkers in isolated perfused rat lungs. Particles were sampled from the studded tire wear of three road pavements containing different rock materials in a road simulator; and from the wear of two brake pad materials using a pin-on-disk machine. Isolated rat lungs inhaled the coarse and fine fractions of the sampled particles resulting in an estimated total particle lung dose of 50 μg. The tidal volume (TV) was measured during the particle exposure and the following 50 min. Perfusate and BALF were analyzed for the cytokines TNF, CXCL1 and CCL3. The TV of lungs exposed to rock materials was significantly reduced after 25 min of exposure compared to the controls, for quartzite already after 4 min. The particles of the heavy-duty brake pads had no effect on the TV. Brake particles resulted in a significant elevation of CXCL1 in the perfusate. Brake particles showed significant elevations of all three measured cytokines, and quartzite showed a significant elevation of TNF in BALF. The study shows that the toxic effect on lungs exposed to airborne particles can be investigated using measurements of tidal volume. Furthermore, the study shows that the choice of rock material in road pavements has the potential to affect the toxicity of road wear PM10.


Mitochondrial and nucleolar localization of cysteine desulfurase Nfs and the scaffold protein Isu in Trypanosoma brucei.
Trypanosoma brucei has a complex life cycle during which its single mitochondrion is subjected to major metabolic and morphological changes. While the procyclic stage (PS) of the insect vector contains a large and reticulated mitochondrion, its counterpart in the bloodstream stage (BS) parasitizing mammals is highly reduced and seems to be devoid of most functions. We show here that key Fe-S cluster assembly proteins are still present and active in this organelle and that produced clusters are incorporated into overexpressed enzymes. Importantly, the cysteine desulfurase Nfs, equipped with the nuclear localization signal, was detected in the nucleolus of both T. brucei life stages. The scaffold protein Isu, an interacting partner of Nfs, was also found to have a dual localization in the mitochondrion and the nucleolus, while frataxin and both ferredoxins are confined to the mitochondrion. Moreover, upon depletion of Isu, cytosolic tRNA thiolation dropped in the PS but not BS parasites.


Identification of polyunsaturated fatty acids as potential biomarkers of osteoarthritis after sodium hyaluronate and mesenchymal stem cell treatment through metabolomics.
Introduction: Osteoarthritis (OA) is a prevalent joint disorder worldwide. Sodium hyaluronate (SH) and mesenchymal stem cells (MSCs) are promising therapeutic strategies for OA. Previous studies showed they could improve knee function and clinical symptoms of OA. However, the mechanism of the therapeutic effects on the improvement of OA has not been clearly explained. Methods: In our study, we used a technique called 5-(diisopropylamino)amylamine derivatization liquid chromatography coupled with mass spectrometry to find the metabolites in OA synovial fluid under different treatments. Results and Discussion: After looking into the metabolomics, we discovered that SH and MSC treatment led to the downregulation of ω-6 polyunsaturated fatty acids (PUFAs) and the upregulation of ω-3 PUFAs. Significantly, the contents of 5(S)-HETE, PGA2, PGB2, and PGJ2 were lower in the MSC group than in the SH group after quantification using 5-(diisopropylamino)amylamine derivatization-UHPLC-QQQ-MS. This is the first report on the relationship of 11(S)-HETE, PGA2, PGB2, PGF2β, 11β-PGF2α, and DK-PGE2 with OA. Moreover, the correlation analysis of metabolites and inflammation factors showed the positive association of ω-6 PUFAs with pro-inflammation cytokines, and of ω-3 PUFAs with anti-inflammation cytokines. Our results indicated the therapeutic effect of SH and MSCs in patients with OA. In addition, this reliable metabolic approach could uncover novel biomarkers to treat OA.


A single treatment with microcapsules containing a CXCR4 antagonist suppresses pulmonary metastasis of murine melanoma.
Biodegradable poly D,L-lactic acid (PLA, molecular weight: ca. 5000) microcapsules containing a CXCR4 antagonist (4F-benzoyl-TE14011) were prepared (4F-benzoyl-TE14011-PLA), and their anti-metastatic activity was evaluated in mice. A single subcutaneous administration of 4F-benzoyl-TE14011-PLA significantly reduced the number of colonies formed by pulmonary metastasis of B16-BL6 melanoma cells expressing CXCR4. The same dose of 4F-benzoyl-TE14011 in a single or a series of treatments affected little. The substance 4F-benzoyl-TE14011 dose-dependently suppressed B16-BL6 cell growth. In the cells cultured with SDF-1, a more potent suppression was observed. 4F-Benzoyl-TE14011 was rapidly released from 4F-benzoyl-TE14011-PLA for an initial period, both in vitro and in vivo. A steady release was thereafter observed. Therefore, this drug release profile might contribute to prevention of melanoma metastasis at the steps involving the migration and cell growth. These results also show that a sustained drug release formulation could be a useful drug delivery system for CXCR4 antagonists.


Magnetic Resonance vs. Intraoral Ultrasonography in the Preoperative Assessment of Oral Squamous Cell Carcinoma: A Systematic Review and Meta-Analysis.
Background: Preoperative assessment is critical to decide the most adequate surgical strategy for oral squamous cell carcinoma (SCC). Magnetic resonance (MR) and intraoral ultrasonography (US) have been reported to be of great value for preoperative estimation of depth of invasion (DOI) and/or tumor thickness (TT). This review aims to analyze the accuracy of MR and intraoral US in determining DOI/TT in oral SCC, by assuming histological evaluation as the reference method. Methods: The procedure was conducted following the modified 2009 Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. We performed a systematic search of papers on PubMed, Scopus, Web of Science, and Cochrane Library databases until July 31st, 2019. For quantitative synthesis, we included nine studies (487 patients) focused on MR, and 12 (520 patients) focused on intraoral US. The Pearson correlation coefficient (r) between DOI/TT evaluated with MR or intraoral US was assumed as effect size. A meta-analysis (MA) for each study group (MR and US) was performed by using the random-effects models with the DerSimonian-Laird estimator and r-to-z transformation. Results: In the MA for MR studies, a high heterogeneity was found (I 2 = 94.84%; Q = 154.915, P < 0.001). No significant risk of bias occurred by evaluating funnel plot asymmetry (P = 0.563). The pooled (overall) r of the MR studies was 0.87 (95% CI from 0.82 to 0.92), whereas the pooled r-to-z transformed was 1.44 (95% CI from 1.02 to 1.85). In the MA for US studies a high heterogeneity was found (I 2 = 93.56%; Q = 170.884, P < 0.001). However, no significant risk of bias occurred (P = 0.779). The pooled r of the US studies was 0.96 (95% CI from 0.94 to 0.97), whereas the pooled r-to-z transformed was 1.76 (95% CI from 1.39 to 2.13). These outputs were confirmed in additional MA performed by enrolling only MR (n = 8) and US (n = 11) studies that evaluated TT. Conclusions: MR and intraoral US seem to be promising approaches for preoperative assessment of DOI/TT in oral SCC. Remarkably, a higher pooled r and r-to-z transformed were observed in the intraoral US studies, suggesting that this approach could be more closely related to histopathological findings.


Morphological analyses of the human tongue musculature for three-dimensional modeling.
Skilled movements of the tongue in speech articulation reflect complex formation of the tongue musculature, although its description in the anatomical literature is rather limited for developing a realistic computational model of the tongue. This study presents detailed descriptions of the muscular structure of the human tongue based on macroscopic and microscopic observations and provides three-dimensional schemata of the tongue musculature. Histologic examination revealed that the tongue consists of five strata, stacked along the courses of the fibers of the genioglossus muscle in proximal-distal directions. This stratum structure exists in the entire tongue tissue, indicating that the lingual musculature can be divided into the inner and outer regions. The former consisted of the "stem" and "core," and the latter of the "cover" and "fringe." In gross dissection, the tongue was cut into wedge-like blocks along the course of the genioglossus muscle to examine muscle fiber arrangement. Using this approach, it was determined that serial repetitions of "structural units" composed the inner musculature of the tongue. Each unit consisted of a pair of thin muscle fiber laminae; one was composed of the genioglossus and vertical muscles, and the other of the transverse muscle. In the apex, the laminae lacked the fibers of the genioglossus. These findings have been incorporated in three-dimensional schemata of the tongue musculature.


Abomasal infusion of ground corn and ammonium chloride in early-lactating Holstein-Friesian dairy cows to induce hindgut and metabolic acidosis.
Next to rumen acidosis, other forms of acidosis may also affect lactational performance of cows. Therefore, the effects of hindgut acidosis, induced via abomasal infusion of ground corn, and metabolic acidosis, induced via abomasal infusion of NH4Cl, were studied in cows in early lactation. Observations were made on intake and digestibility of nutrients, lactation performance, energy and N partitioning, blood acid-base status, and rumen and hindgut fermentation characteristics. In a 6 × 6 Latin square design, 6 rumen-fistulated, second-lactation Holstein-Friesian dairy cows (48 ± 17 d in milk) were subjected to 5 d of continuous abomasal infusions of water as control, or solutions of 2.5 mol of NH4Cl/d, 5.0 mol of NH4Cl/d, 3.0 kg of ground corn/d, or the combination of ground corn with either of the 2 NH4Cl levels, followed by 2 d of rest. Treatment solutions were administered via peristaltic pumps through infusion lines attached to the rumen cannula plug and an abomasal infusion line with a flexible disk (equipped with holes to allow digesta passage) to secure its placement through the sulcus omasi. A total mixed ration consisting of 70% grass silage and 30% concentrate (on dry matter basis) was fed at 95% of ad libitum intake of individual cows. The experiment was conducted in climate respiration chambers to determine feed intake, lactation performance, and energy and N balance. Abomasal infusion of NH4Cl affected the acid-base status of the cows, but more strongly when in combination with abomasal infusion of ground corn. Metabolic acidosis (defined as a blood pH < 7.40, blood HCO3 concentration < 25.0 mmol/L, and a negative base excess) was observed with 5.0 mol of NH4Cl/d, 3.0 kg of ground corn/d + 2.5 mol of NH4Cl/d, and 3.0 kg of ground corn/d + 5.0 mol of NH4Cl/d. Metabolic acidosis was associated with decreased milk lactose content, metabolic body weight, energy retained as protein, and fecal N excretion, and increased urine N excretion, and tended to decrease intake of nutrients. Digestibility of several nutrients increased with 5.0 mol of NH4Cl/d, likely as a result of decreased intake. Abomasal ground corn infusion resulted in hindgut acidosis, where fecal pH decreased from 6.86 without ground corn to 6.00 with ground corn, regardless of NH4Cl level. The decrease in fecal pH was likely the result of increased hindgut fermentation, evidenced by increased fecal volatile fatty acid concentrations. Hindgut acidosis was associated with decreased digestibility of nutrients, except for starch, which increased, and crude fat, which was not affected. No systemic inflammatory response was observed, suggesting that the hindgut epithelium was not severely affected by the more acidic conditions or barrier damage. Abomasal infusion of ground corn increased milk yield, milk protein and lactose yield, fecal N excretion, N use efficiency, and total energy retained as well as energy retained in fat, and reduced milk fat content and urine N excretion.


Energy taxis is the dominant behavior in Azospirillum brasilense.
Energy taxis encompasses aerotaxis, phototaxis, redox taxis, taxis to alternative electron acceptors, and chemotaxis to oxidizable substrates. The signal for this type of behavior is originated within the electron transport system. Energy taxis was demonstrated, as a part of an overall behavior, in several microbial species, but it did not appear as the dominant determinant in any of them. In this study, we show that most behavioral responses proceed through this mechanism in the alpha-proteobacterium Azospirillum brasilense. First, chemotaxis to most chemoeffectors typical of the azospirilla habitat was found to be metabolism dependent and required a functional electron transport system. Second, other energy-related responses, such as aerotaxis, redox taxis, and taxis to alternative electron acceptors, were found in A. brasilense. Finally, a mutant lacking a cytochrome c oxidase of the cbb(3) type was affected in chemotaxis, redox taxis, and aerotaxis. Altogether, the results indicate that behavioral responses to most stimuli in A. brasilense are triggered by changes in the electron transport system.


A Voxel Vascular Structure-based Mannequin-like Arm Electromagnetic Model for Radio Frequency Biomedical Sensors.
Radio Frequency (RF) sensor is widely used to monitor physiological signals. Generally, RF sensor simulation is mostly done using a layered model, which sometimes cannot model the accurate properties in the real world. A voxel vascular structure-based mannequin-like arm electromagnetic model (VVS-MaM) is proposed to evaluate the RF sensor, which mainly gathers the real physiological signal. This model is built with high-precision Magnetic Resonance Imaging (MRI), and it can finish fast simulation while there is also a voxel-like part in it which means it has the advantages of both the layered model and the real human model. After modelling, both simulation and in-vivo experiments are designed to test this sensor. In the simulation, the simulated standard resonant frequency of the equivalent model is 1.8137 GHz, and the relative error of the VVS-MaM is 0.012 GHz, which is closer to the standard value than the layer model result of 0.049 GHz. Meanwhile, in the in-vivo experiments, an RF sensor based on a composite right/left-handed transmission line (CRLH-TL) and complementary split resonator rings (CSRRs) are fabricated, and the measurements from the real experiments are gathered and stored to compare with that of the simulation. The comparison shows that the relative error of the VVS-MaM (0.08804 GHz)is closer to the in-vivo measurements than that of the layer model (0.09891 GHz), which validates the performance of VVS-MaM.Clinical Relevance-Radio Frequency, magnetic resonance imaging, scattering parameter, composite right/left-handed, complementary split resonator ring.


Allotypes of murine factor B controlled by a locus within the S region of the H-2 complex.
Bf protein was directly precipitated from a mixture of EDTA-plasma from 13 different, inbred strains and rabbit IgG anti-mouse Bf, and was isolated by SDS-PAGE. The gel pieces containing Bf protein were digested by trypsin after labeling with 125I-Na, and then peptides of Bf protein with each mouse strain were compared by two-dimensional peptide mapping. The results for the peptide patterns with the standard B10 congenic strains (all of which Bf phenotypes had been already designated as Bf.1 because of the identical isoelectric point values) revealed two distinct peptide patterns. The peptide pattern of Bf protein appeared to be identical in each of the inbred strains, but only one of the 36 spots in the fingerprint observed either with B10 or B10.BR (designated tentatively as Bf.1 (b.k.) ) was not detected in that from B10.D2 and B10.S (designated tentatively as Bf.1 (d.s.) ). The results of peptide mapping with intra-H-2 recombinant inbred strains show that this structural variant was mapped to the S region, which is direct evidence that allotypes of murine Bf are encoded by a structural gene within the MHC.


Sensory neuropathy with bone destruction due to a mutation in the membrane-shaping atlastin GTPase 3.
Many neurodegenerative disorders present with sensory loss. In the group of hereditary sensory and autonomic neuropathies loss of nociception is one of the disease hallmarks. To determine underlying factors of sensory neurodegeneration we performed whole-exome sequencing in affected individuals with the disorder. In a family with sensory neuropathy with loss of pain perception and destruction of the pedal skeleton we report a missense mutation in a highly conserved amino acid residue of atlastin GTPase 3 (ATL3), an endoplasmic reticulum-shaping GTPase. The same mutation (p.Tyr192Cys) was identified in a second family with similar clinical outcome by screening a large cohort of 115 patients with hereditary sensory and autonomic neuropathies. Both families show an autosomal dominant pattern of inheritance and the mutation segregates with complete penetrance. ATL3 is a paralogue of ATL1, a membrane curvature-generating molecule that is involved in spastic paraplegia and hereditary sensory neuropathy. ATL3 proteins are enriched in three-way junctions, branch points of the endoplasmic reticulum that connect membranous tubules to a continuous network. Mutant ATL3 p.Tyr192Cys fails to localize to branch points, but instead disrupts the structure of the tubular endoplasmic reticulum, suggesting that the mutation exerts a dominant-negative effect. Identification of ATL3 as novel disease-associated gene exemplifies that long-term sensory neuronal maintenance critically depends on the structural organisation of the endoplasmic reticulum. It emphasizes that alterations in membrane shaping-proteins are one of the major emerging pathways in axonal degeneration and suggests that this group of molecules should be considered in neuroprotective strategies.


Cascaded diffraction in optical systems. Part II: example calculations.
Real imaging optical systems are in any case finite in diameter and therefore limit the ray cones coming from the various object points. The truncation of the ray cones generates edge diffraction effects, which have an impact on the point spread function and therefore on the resolution of the system. Traditionally real calculations of the diffraction effects are mostly simplified by assuming a truncation in the exit pupil only. In this hybrid approach, the light propagation therefore can be calculated by raytrace first, and the more cumbersome diffraction calculated is only performed between exit pupil and image plane. To come to a better quantitative understanding of the validity of this approximation, a more refined model is developed in an adjacent publication. Some example calculations for real setups and an overview about the order of magnitude of the cascaded diffraction effects are worked out in this contribution.


Genome-Wide Identification and Transcriptomic Analysis of MicroRNAs Across Various Amphioxus Organs Using Deep Sequencing.
Amphioxus is the closest living invertebrate proxy of the vertebrate ancestor. Systematic gene identification and expression profile analysis of amphioxus organs are thus important for clarifying the molecular mechanisms of organ function formation and further understanding the evolutionary origin of organs and genes in vertebrates. The precise regulation of microRNAs (miRNAs) is crucial for the functional specification and differentiation of organs. In particular, those miRNAs that are expressed specifically in organs (OSMs) play key roles in organ identity, differentiation, and function. In this study, the genome-wide miRNA transcriptome was analyzed in eight organs of adult amphioxus Branchiostoma belcheri using deep sequencing. A total of 167 known miRNAs and 23 novel miRNAs (named novel_mir), including 139 conserved miRNAs, were discovered, and 79 of these were identified as OSMs. Additionally, analyses of the expression patterns of eight randomly selected known miRNAs demonstrated the accuracy of the miRNA deep sequencing that was used in this study. Furthermore, potentially OSM-regulated genes were predicted for each organ type. Functional enrichment of these predicted targets, as well as further functional analyses of known OSMs, was conducted. We found that the OSMs were potentially to be involved in organ-specific functions, such as epidermis development, gonad development, muscle cell development, proteolysis, lipid metabolism, and generation of neurons. Moreover, OSMs with non-organ-specific functions were detected and primarily include those related to innate immunity and response to stimuli. These findings provide insights into the regulatory roles of OSMs in various amphioxus organs.


A newly detected class of mammalian single strand-specific DNA-binding proteins. Effects on DNA polymerase alpha-catalyzed DNA synthesis.
Using a rapid purification scheme, we have isolated from calf thymus cells two single strand-specific DNA-binding proteins, with apparent molecular masses of 48 and 61 kDa. These proteins remained undetected in earlier studies on DNA-binding proteins from calf thymus because they are extremely sensitive to proteolytic breakdown occurring during purification. The proteins are immunologically and biochemically related. They also share a number of functional properties. Both proteins bind noncooperatively to single-stranded DNA and almost totally ignore double-stranded DNA. Both proteins stimulate DNA synthesis catalyzed by mammalian DNA polymerase alpha in the presence of an excess of "activated" DNA as primer-template. The stimulation factor was between 2 and 100 depending on the protein/DNA ratio. We have analyzed this effect in more detail with specifically primed single-stranded phage fd DNA as template and concluded that the proteins block nonproductive polymerase-binding sites on single-stranded DNA sequences, thereby increasing the probability for an association of polymerase with 3'-OH primer ends.


Use of different D-dimer levels to exclude venous thromboembolism depending on clinical pretest probability.
Currently, the same D-dimer cut-off point is used to define a positive result for all patients with suspected venous thromboembolism, regardless of their pretest probability. However, use of a relatively high D-dimer cut-off point (lower sensitivity) for those with a low clinical pretest probability, and a low D-dimer cut-off point (higher sensitivity) for those with a high clinical pretest probability, may be preferable. To determine if using three different D-dimer cut-off points according to low, moderate or high clinical pretest probability has greater utility for exclusion of venous thromboembolism than using the same single D-dimer cut-off point in all patients. Data from a previously published study of 571 patients was used to identify the highest D-dimer cut-off point with a negative predictive value of at least 98% for the subgroup of patients with low and high pretest probability. The D-dimer cut-off point for those with moderate clinical pretest probability remained unchanged [0.5 fibrinogen equivalent units (FEU) microgram mL(-1)]. Accuracy of D-dimer testing for venous thromboembolism using three cut-off points vs. one cut-off point was than determined. D-dimer cut-off points of 0.2 and 2.1 FEU microgram mL(-1) were selected for the high and low pretest probability groups, respectively. When three pretest probability-specific cut-off points were used instead of the previously determined single D-dimer cut-off point (0.5 FEU microgram mL(-1)), sensitivity and negative predictive value were unchanged (95 and 98%, respectively), but specificity increased from 44.7 to 60.4% (P < 0.001). This resulted in exclusion of venous thromboembolism in 80 additional patients. Use of three pretest probability-specific D-dimer cut-off points rather than a single D-dimer cut-off point for all patients, has the potential to increase the utility of D-dimer testing for the diagnosis of venous thromboembolism.


Psychological change mechanisms in anorexia nervosa treatments: how much do we know?
Anorexia nervosa (AN) is a debilitating and often chronic and treatment-resistant disorder. Despite decades of theoretical progress and research, many questions remain with regard to the psychological mechanisms explaining why and how some AN patients respond to treatment whereas others do not. Based on the premise that the broader, noneating disorders psychotherapy research literature, and particularly the common factors literature, can inform AN treatment development efforts, we review a set of selected psychological change mechanisms and describe how they might be relevant in the context of AN treatment response. Specifically, we suggest that a systematic consideration of constructs such as basic psychological needs, expectancies, the therapeutic alliance, experiential avoidance, and patient motivation for change might help illuminate how patients do or do not benefit from AN treatment. We briefly describe an ongoing multicenter trial in which the constructs introduced here are being measured on a weekly basis and are examined as potential mediators of treatment response. The article aims to contribute to the AN literature by introducing a set of potentially important change constructs that we think ought to be studied in greater depth by AN researchers.


Comparison of three medication adherence measures in patients taking warfarin.
In order to improve warfarin adherence, we must first know the rate of non-adherence and the reasons for it. Assessment of warfarin adherence is important in improving patients' warfarin-taking behavior and International Normalized Ratio (INR) control. This study aimed to compare three medication adherence measures in patients taking warfarin: the 8-item Morisky Medication Adherence Scale (MMAS), the 100-point Visual Analogue Scale (VAS) and medication refill adherence (MRA). A cross-sectional observational study was conducted in a convenience sample of 174 patients taking warfarin at an anticoagulation clinic. A survey questionnaire that included the MMAS, VAS, and demographic and warfarin-related clinical questions was administered to patients in English or Chinese depending on their preferences. The MRA and INR values were retrieved from hospital electronic databases. Most participants were adherent to warfarin as indicated by the three measures. A weak association was found between the MMAS and the other two adherence measures. The MMAS and MRA were associated with the percentage of INRs within range in the past 2 weeks. Moreover, the MRA was weakly associated with time within the therapeutic INR range in the past 3 months and 2 weeks. The findings provide insights into the differences among three medication adherence measures and may assist healthcare providers to select the most suitable measure for the assessment of warfarin adherence.


"Punched in the Balls": Male Intimate Partner Violence Disclosures and Replies on Reddit.
Research on male intimate partner violence (IPV) survivors is limited. The sparse research on male IPV disclosure suggest males receive more negative and less helpful responses from potential sources of formal or informal support. Males may seek support on social networking sites (SNSs), hence, it is important to understand their emerging experiences of virtual disclosures. This study examined the nature and content of responses to IPV disclosures by male IPV survivors on a popular SNS (reddit.com). Search of Reddit submissions related to male IPV were carried out using three IPV related keywords for the calendar month of February 2019, resulting in 917 submissions. Twelve submissions that focused on male IPV disclosure were examined in detail. The 12 submissions were analyzed using quantitative content analysis while associated comments (n = 569) were analyzed using qualitative approach. Two-thirds of the disclosures (8/12) were of personal IPV experiences. All disclosure narratives identified the sex of perpetrator, most stated the types of abuse (7/12), and some revealed the outcomes of past disclosures (4/12). Six major themes were developed through qualitative analysis of the associated comments: (1) Informational Support, (2) Nurturant Support, (3) Tangible Aid, (4) Negative Response (5) Self-Defence, and (6) Reciprocal Disclosure. Overall, males experienced a majority of supportive responses to IPV disclosures and some negative responses including criticism and minimizing the abuse. Males take risks in disclosure of IPV in person and online.


Vaxign-DL: A Deep Learning-based Method for Vaccine Design and its Evaluation.
Reverse vaccinology (RV) provides a systematic approach to identifying potential vaccine candidates based on protein sequences. The integration of machine learning (ML) into this process has greatly enhanced our ability to predict viable vaccine candidates from these sequences. We have previously developed a Vaxign-ML program based on the eXtreme Gradient Boosting (XGBoost). In this study, we further extend our work to develop a Vaxign-DL program based on deep learning techniques. Deep neural networks assemble non-linear models and learn multilevel abstraction of data using hierarchically structured layers, offering a data-driven approach in computational design models. Vaxign-DL uses a three-layer fully connected neural network model. Using the same bacterial vaccine candidate training data as used in Vaxign-ML development, Vaxign-DL was able to achieve an Area Under the Receiver Operating Characteristic of 0.94, specificity of 0.99, sensitivity of 0.74, and accuracy of 0.96. Using the Leave-One-Pathogen-Out Validation (LOPOV) method, Vaxign-DL was able to predict vaccine candidates for 10 pathogens. Our benchmark study shows that Vaxign-DL achieved comparable results with Vaxign-ML in most cases, and our method outperforms Vaxi-DL in the accurate prediction of bacterial protective antigens.


Alcoholic cardiomyopathy: incidence, clinical characteristics, and pathophysiology.
In the United States, in both sexes and all races, long-term heavy alcohol consumption (of any beverage type) is the leading cause of a nonischemic, dilated cardiomyopathy, herein referred to as alcoholic cardiomyopathy (ACM). ACM is a specific heart muscle disease of a known cause that occurs in two stages: an asymptomatic stage and a symptomatic stage. In general, alcoholic patients consuming > 90 g of alcohol a day (approximately seven to eight standard drinks per day) for > 5 years are at risk for the development of asymptomatic ACM. Those who continue to drink may become symptomatic and develop signs and symptoms of heart failure. ACM is characterized by an increase in myocardial mass, dilation of the ventricles, and wall thinning. Changes in ventricular function may depend on the stage, in that asymptomatic ACM is associated with diastolic dysfunction, whereas systolic dysfunction is a common finding in symptomatic ACM patients. The pathophysiology of ACM is complex and may involve cell death (possibly due to apoptosis) and changes in many aspects of myocyte function. ACM remains an important cause of a dilated cardiomyopathy, and in latter stages can lead to heart failure. Alcohol abstinence, as well as the use of specific heart failure pharmacotherapies, is critical in improving ventricular function and outcomes in these patients.


In vivo activation by ethanol of plasma membrane ATPase of Saccharomyces cerevisiae.
Ethanol, in concentrations that affect growth and fermentation rates (3 to 10% [vol/vol]), activated in vivo the plasma membrane ATPase of Saccharomyces cerevisiae. The maximal value for this activated enzyme in cells grown with 6 to 8% (vol/vol) ethanol was three times higher than the basal level (in cells grown in the absence of ethanol). The Km values for ATP, the pH profiles, and the sensitivities to orthovanadate of the activated and the basal plasma membrane ATPases were virtually identical. A near-equivalent activation was also observed when cells grown in the absence of ethanol were incubated for 15 min in the growth medium with ethanol. The activated state was preserved after the extraction from the cells of the membrane fraction, and cycloheximide appeared to prevent this in vivo activation. After ethanol removal, the rapid in vivo reversion of ATPase activation was observed. While inducing the in vivo activation of plasma membrane ATPase, concentrations of ethanol equal to and greater than 3% (vol/vol) also inhibited this enzyme in vitro. The possible role of the in vivo activation of the plasma membrane proton-pumping ATPase in the development of ethanol tolerance by this fermenting yeast was discussed.


Base Editors: Modular Tools for the Introduction of Point Mutations in Living Cells.
Base editors are a new family of programmable genome editing tools that fuse ssDNA (single stranded DNA) modifying enzymes to catalytically inactive CRISPR-associated (Cas) endonucleases to induce highly efficient single base changes. With dozens of base editors now reported, it is apparent that these tools are highly modular; many combinations of ssDNA modifying enzymes and Cas proteins have resulted in a variety of base editors, each with its own unique properties and potential uses. In this perspective, we describe currently available base editors, highlighting their modular nature and describing the various options available for each component. Furthermore, we briefly discuss applications in synthetic biology and genome engineering where base editors have presented unique advantages over alternative techniques.


Asthma, severe acute respiratory syndrome coronavirus-2 and coronavirus disease 2019.
In December 2019, a novel respiratory illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first described and named coronavirus disease 2019 (COVID-19). Although the knowledge base surrounding COVID-19 and SARS-CoV-2 has grown rapidly, significant gaps in our knowledge remain and inaccurate information continues to circulate. This review will discuss the interaction between asthma and COVID-19 to provide a comprehensive understanding based on the currently available published data. Non-SARS human coronaviruses (HCoVs) are a significant cause of asthma exacerbations, but SARS-CoV-2 does not appear to exacerbate asthma. Data thus far strongly suggest that patients with asthma are at no increased risk of infection with SARS-CoV-2 or more severe disease if infected with COVID-19. Although the data are extremely limited on inhaled corticosteroids and biologic medications, there remain no data suggesting that these therapeutics positively or negatively impact the severity or outcome of COVID-19. Data are rapidly evolving regarding COVID-19 and asthma. At this time, asthma does not appear to positively or negatively affect outcomes of COVID-19; however, it is imperative that practitioners keep abreast of the changing literature as we await a vaccine and control of this pandemic.


Role of Ash1l in Tourette syndrome and other neurodevelopmental disorders.
Ash1l potentially contributes to neurodevelopmental diseases. Although specific Ash1l mutations are rare, they have led to informative studies in animal models that may bring therapeutic advances. Ash1l is highly expressed in the brain and correlates with the neuropathology of Tourette syndrome (TS), autism spectrum disorder, and intellectual disability during development, implicating shared epigenetic factors and overlapping neuropathological mechanisms. Functional convergence of Ash1l generated several significant signaling pathways: chromatin remodeling and transcriptional regulation, protein synthesis and cellular metabolism, and synapse development and function. Here, we systematically review the literature on Ash1l, including its discovery, expression, function, regulation, implication in the nervous system, signaling pathway, mutations, and putative involvement in TS and other neurodevelopmental traits. Such findings highlight Ash1l pleiotropy and the necessity of transcending a single gene to complicated mechanisms of network convergence underlying these diseases. With the progress in functional genomic analysis (highlighted in this review), and although the importance and necessity of Ash1l becomes increasingly apparent in the medical field, further research is required to discover the precise function and molecular regulatory mechanisms related to Ash1l. Thus, a new perspective is proposed for basic scientific research and clinical interventions for cross-disorder diseases.


Parenting style, religiosity, peers, and adolescent heavy drinking.
The purpose of this research was to examine whether authoritative, authoritarian, indulgent, and neglectful parenting styles were associated with adolescent alcohol use and heavy drinking, after controlling for peer use, religiosity, and other relevant variables. Structural equation modeling was used to estimate direct and indirect associations of parenting style with alcohol use and heavy drinking among 4,983 adolescents in Grades 7-12. Adolescents whose parents were authoritative were less likely to drink heavily than adolescents from the other three parenting styles, and they were less likely to have close friends who used alcohol. In addition, religiosity was negatively associated with heavy drinking after controlling for other relevant variables. Authoritative parenting appears to have both direct and indirect associations with the risk of heavy drinking among adolescents. Authoritative parenting, where monitoring and support are above average, might help deter adolescents from heavy alcohol use, even when adolescents have friends who drink. In addition, the data suggest that the adolescent's choice of friends may be an intervening variable that helps explain the negative association between authoritative parenting and adolescent heavy drinking.


C-C motif chemokine receptor 9 regulates obesity-induced insulin resistance via inflammation of the small intestine in mice.
Accumulation of adipose tissue macrophages is considered pivotal in the development of obesity-associated inflammation and insulin resistance. In addition, recent studies suggest an involvement of the intestine as the primary organ in inducing hyperglycaemia and insulin resistance. We have reported that the C-C motif chemokine receptor (CCR) CCR9 is associated with intestinal immunity and has a pathogenic role in various liver diseases. However, its contribution to type 2 diabetes is unknown. In the current study, we aimed to clarify the involvement of CCR9 in the pathology of type 2 diabetes and the potential underlying mechanisms. To elucidate how CCR9 affects the development of metabolic phenotypes, we examined the impact of CCR9 deficiency on the pathogenesis of type 2 diabetes using male C57BL/6J (wild-type [WT]) and CCR9-deficient (CCR9 knockout [KO]) mice fed a 60% high-fat diet (HFD) for 12 weeks. WT and Ccr9KO mice fed an HFD exhibited a comparable weight gain; however, glucose tolerance and insulin resistance were significantly improved in Ccr9KO mice. Moreover, visceral adipose tissue (VAT) and the liver of Ccr9KO mice presented with less inflammation and increased expression of glucose metabolism-related genes than WT mice. Ccr9 and Ccl25 expression were specifically higher in the small intestine but was not altered by HFD feeding and type 2 diabetes development. Accumulation of IFN-γ-producing CD4+ T lymphocytes and increased intestinal permeability in the small intestine was observed in WT mice following HFD feeding, but these changes were suppressed in HFD-fed Ccr9KO mice. Adoptive transfer of gut-tropic CCR9-expressing T lymphocytes partially reversed the favourable glucose tolerance found in Ccr9KO mice via exacerbated inflammation in the small intestine and VAT. CCR9 plays a central role in the pathogenesis of type 2 diabetes by inducing an inflammatory shift in the small intestine. Our findings support CCR9 as a new therapeutic target for type 2 diabetes via the gut-VAT-liver axis.


Mode-specific strategy for controlling a single-molecule reaction.
Inelastic tunnelling electrons are a proper excitation source to induce chemical transformations on a single adsorbate. When their energy is tuned to that of molecular vibrational states, the modification may follow complex internal vibrational pathways. Here, we analyse our recent results on the selective excitation of ammonium stretching or bending modes to control the outcome of a simple bond-cleavage reaction. With the help of model calculations, we provide a detailed molecular-scale picture of the competing internal pathways leading to molecular movement. A mode-selective strategy, based on local excitations of specific reaction coordinates, has an important drawback when applied to adsorbate systems due to the problem of fast energy randomization. The success of such a mode-selective strategy is determined here by the ability of the scanning tunnelling microscope to study reactivity in the limit of very low yield and very low power irradiation, in a regime where vibrational heating of the adsorbate/surface system becomes negligible.


AICAR activates the pluripotency transcriptional network in embryonic stem cells and induces KLF4 and KLF2 expression in fibroblasts.
Pluripotency, the property of a cell to differentiate into all cellular types of a given organism, is central to the development of stem cell-based therapies and regenerative medicine. Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4. Recently, it has been shown that pluripotency can be induced in somatic cells by viral-mediated expression of the transcription factors Oct3/4, Sox2, Klf4, and c-Myc. Here we show that 5-Aminoimidazole-4-carboxamide-1-b-riboside (AICAR) is able to activate the molecular circuitry of pluripotency in mouse embryonic stem cells (mESC) and maintain Nanog and Oct4 expression in mESC exposed to the differentiating agent retinoic acid. We also show that AICAR is able to induce Klf4, Klf2 and Myc expression in both mESC and murine fibroblasts. AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells. AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency.


An integrated analysis of match-related fatigue in professional rugby league.
This study examined the changes in external outputs, including metabolic power variables, and internal response whilst considering contextual factors on physical performance variables during rugby league match play. Physical performance (total distance, high-speed running and high-power distances, average metabolic power), heart-rate (percentage heart-rate peak and training impulse), collisions (attacking and defensive) and contextual (time in attack, time in defence, time out of play) data were collected from 18 rugby league players during 38 games throughout two National Rugby League seasons. Physical variables were highest in the first 10-min period of each half (P < 0.001). Heart-rate indices peaked in the second 10-min period and were lower during second half periods (P < 0.001). Few differences existed in collisions and contextual factors across 10-min periods. Physical variables were highest during the first 5-min period compared to the final (P < 0.001). There was no difference in heart-rate response, attacking collisions or contextual factors between these periods. Following the peak 5-min period in the match, there were reductions in physical, heart-rate, defensive collisions and contextual factors (P < 0.001). The data show temporal changes in physical performance, heart-rate response and collisions during rugby league match play, although these are affected by contextual factors.


Transit peptide elements mediate selective protein targeting to two different types of chloroplasts in the single-cell C4 species Bienertia sinuspersici.
Bienertia sinuspersici is a terrestrial plant that performs C4 photosynthesis within individual cells through operating a carbon concentrating mechanism between different subcellular domains including two types of chloroplasts. It is currently unknown how differentiation of two highly specialized chloroplasts within the same cell occurs as no similar cases have been reported. Here we show that this differentiation in photosynthetic cells of B. sinuspersici is enabled by a transit peptide (TP) mediated selective protein targeting mechanism. Mutations in the TPs cause loss of selectivity but not general loss of chloroplast import, indicating the mechanism operates by specifically blocking protein accumulation in one chloroplast type. Hybrid studies indicate that this selectivity is transferable to transit peptides of plants which perform C4 by cooperative function of chloroplasts between two photosynthetic cells. Codon swap experiments as well as introducing an artificial bait mRNA show that RNA affects are not crucial for the sorting process. In summary, our analysis shows how the mechanism of subcellular targeting to form two types of chloroplast within the same cell can be achieved. This information is not only crucial for understanding single-cell C4 photosynthesis; it provides new insights in control of subcellular protein targeting in cell biology.


Leakage beyond the primary lesion: A temporal analysis of cerebrovascular dysregulation at sites of hippocampal secondary neurodegeneration following cortical photothrombotic stroke.
We have previously demonstrated that a cortical stroke causes persistent impairment of hippocampal-dependent cognitive tasks concomitant with secondary neurodegenerative processes such as amyloid-β accumulation in the hippocampus, a region remote from the primary infarct. Interestingly, there is emerging evidence suggesting that deposition of amyloid-β around cerebral vessels may lead to cerebrovascular structural changes, neurovascular dysfunction, and disruption of blood-brain barrier integrity. However, there is limited knowledge about the temporal changes of hippocampal cerebrovasculature after cortical stroke. In the current study, we aimed to characterise the spatiotemporal cerebrovascular changes after cortical stroke. This was done using the photothrombotic stroke model targeting the motor and somatosensory cortices of mice. Cerebrovascular morphology as well as the co-localisation of amyloid-β with vasculature and blood-brain barrier integrity were assessed in the cortex and hippocampal regions at 7, 28 and 84 days post-stroke. Our findings showed transient cerebrovascular remodelling in the peri-infarct area up to 28 days post-stroke. Importantly, the cerebrovascular changes were extended beyond the peri-infarct region to the ipsilateral hippocampus and were sustained out to 84 days post-stroke. When investigating vessel diameter, we showed a decrease at 84 days in the peri-infarct and CA1 regions that were exacerbated in vessels with amyloid-β deposition. Lastly, we showed sustained vascular leakage in the peri-infarct and ipsilateral hippocampus, indicative of a compromised blood-brain-barrier. Our findings indicate that hippocampal vasculature may represent an important therapeutic target to mitigate the progression of post-stroke cognitive impairment.


Decrease in delta and mu opioid receptor binding capacity in rat brain after chronic etorphine treatment.
The modulatory effect of continued activation of opiate receptors with agonist on the receptor level was examined in current studies. Rats were rendered tolerant to etorphine by s.c. implantation of osmotic minipumps containing 3 mg/ml of etorphine for up to 7 days. During this period, there were a time-dependent increase in the AD50 values of etorphine to inhibit the tail-flick response and an increase in naloxone-precipitated withdrawal signs. When these animals and others were sacrificed and the opiate receptor binding properties were examined, it could be demonstrated that there was also a time-dependent decrease in the amount of [3H]diprenorphine specifically bound, with maximal attenuation reached 3 days after implantation. There was no alteration in alpha 2 adrenergic receptor binding. This observed decrease in [3H]diprenorphine binding was not due to the presence of nonwashed etorphine in the membrane, for acute administration of the same dose of etorphine before sacrificing did not produce an attenuation of the opiate receptor binding. Further examination of opiate receptor binding in the brain regions of cortex, midbrain and striatum revealed the greatest decrease in the amount of [3H]diprenorphine bound in striatal region after chronic etorphine treatment, a 68% decrease. When the relative decrease in mu and delta opioid receptor binding was determined by carrying out [3H]-D-Ala2, DLeu5-enkephalin binding in the presence of 1 microM morphiceptin, it was observed that, 3 days after etorphine treatment, there was a decrease in mu opioid receptor binding, with minimal change in delta opioid receptor binding in both brain regions of striatum and midbrain. Only in cortex was a decrease in binding of both receptor subtypes observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Transantral endoscopic orbital floor exploration: a cadaver and clinical study.
A cadaver and clinical study was performed to determine the value of transantral endoscopy in diagnosis and treatment of orbital floor fractures. Six fresh cadaver heads were dissected using a 30 degree, 4-mm endoscope through a 1 cm2 antrotomy. In the cadaver, the orbital floor and the course of the infraorbital nerve were easily identified. The infraorbital nerve serves as a reference point for evaluation of fracture size; three zones of the floor are described that are oriented relative to the infraorbital nerve. In the clinical study, nine patients with orbital floor fracture initially underwent endoscopy at the time of fracture repair: three patients had comminuted zygomatico-orbital fractures, five had monofragmented tetrapod fractures, and one had an isolated orbital blowout fracture. Endoscopic dissection of the orbital fractures revealed seven fractures with an area > 2 cm2 and two fractures with an area of < 2 cm2. The isolated orbital floor blowout fracture had entrapped periorbital tissue, which was completely reduced endoscopically. A separate patient with a < 2 cm2 displaced fracture also had stable endoscopic reduction. In the remaining seven patients, the endoscopic technique assisted with the floor reconstruction by identifying the precise fracture configuration as well as identifying the stable posterior ledge of the orbital floor fracture. There have been no complications in any of our patients to date. We conclude: (1) Transantral orbital floor exploration allows precise determination of orbital floor fracture size, location, and the presence of entrapped periorbita. The information obtained through endoscopic techniques may be used to select patients who would not benefit from lid approaches to the orbital floor and may possibly eliminate nontherapeutic exploration. (2) Transantral endoscopic orbital floor exploration assists the reduction of complex orbital floor fractures and allows precise identification of the posterior shelf for implant placement. (3) Transantral endoscopic techniques can completely reduce entrapped periorbital tissue caught in a trapdoor type of fracture.


Transmitted drug resistance among antiretroviral-naive patients with established HIV type 1 infection in Santo Domingo, Dominican Republic and review of the Latin American and Caribbean literature.
Emergence of HIV resistance is a concerning consequence of global scale-up of antiretroviral therapy (ART). To date, there is no published information about HIV resistance from the Dominican Republic. The study's aim was to determine the prevalence of transmitted drug resistance (TDR) to reverse transcriptase and protease inhibitors in a sample of chronically HIV-1-infected patients in one clinic in Santo Domingo. The data are presented in the context of a review of the TDR literature from Latin America and the Caribbean. Genotype testing was successfully performed on 103 treatment-naive adults planning to initiate antiretroviral therapy; the World Health Organization (WHO) list of surveillance drug resistance mutations (SDRM) was used to determine the presence of TDR mutations. WHO SDRM were identified in eight patients (7.8%); none had received sdNVP. There were no significant differences in epidemiologic or clinical variables between those with or without WHO SDRM. The prevalence of WHO SDRM was 1.0% and 6.8% for nucleoside reverse transcriptase inhibitors and nonnucleoside reverse transcriptase inhibitors, respectively. No WHO SDRMs for protease inhibitors were identified. Among 12 studies of TDR in the region with a sample size of at least 100 subjects, the reported prevalence of SDRM ranged from 2.8% to 8.1%. The most commonly identified SDRM was K103N. This information adds to our understanding of the epidemiology of TDR in the region and the possible role such mutations could play in undermining first-line treatment. Ongoing surveillance is clearly needed to better understand the TDR phenomenon in the Caribbean.


TSLP and downstream molecules in experimental mouse allergic conjunctivitis.
To explore the potential role of thymic stromal lymphopoietin (TSLP) and its downstream molecules in the development of ocular allergic inflammation using a short ragweed (SRW)-induced mouse model of allergic conjunctivitis (AC). BALB/c mice were topically challenged with SRW pollen after they were sensitized with SRW in the footpad. After the last SRW challenge, the corneal epithelium, conjunctiva, and cervical lymph nodes were harvested for total RNA extraction and gene expression by RT and real-time PCR, and whole eye globes were collected to make cryosections for immunohistochemical staining. Repeated topical challenges with SRW allergen generated typical signs of AC in mice. Compared with the untreated controls, TSLP mRNA expression and immunoreactivity were significantly increased in the corneal and conjunctival epithelia of SRW-induced AC mice. CD11c(+) and OX40L(+) immunoreactive cells largely infiltrated the conjunctiva with increased mRNA levels of CD11c, TSLPR, and OX40L detected in the corneal epithelium, conjunctiva, and cervical lymph nodes. CD4(+) Th2 cell infiltration was evidenced by increased levels of mRNA and immunoreactivity of CD4, IL-4, IL-5, and IL-13 in the ocular surface, mainly in the conjunctiva, accompanied by increased expression of OX40, STAT6, and GATA3, in AC mice. The maturation of immature DCs was observed with the use of TSLP containing conditioned media from corneal epithelial cultures exposed to polyI:C, which stimulates TSLP production. This study provides new findings regarding the role of local mucosal epithelial cells in the initiation of ocular allergic inflammation by producing a novel proallergic cytokine, TSLP, which activates dendritic cells to prime Th2 differentiation and allergic inflammation through the TSLP-TSLPR and OX40L-OX40 signaling pathway.