Medicine by Alexandros G. Sfakianakis,Anapafseos 5 Agios Nikolaos 72100 Crete Greece,00302841026182,00306932607174,alsfakia@gmail.com,
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Παρασκευή 7 Ιουλίου 2017
Reversal learning in C58 mice: Modeling higher order repetitive behavior
Source:Behavioural Brain Research, Volume 332
Author(s): Cristina M. Whitehouse, Lisa S. Curry-Pochy, Robin Shafer, Joseph Rudy, Mark H. Lewis
Restricted, repetitive behaviors are diagnostic for autism and prevalent in other neurodevelopmental disorders. These behaviors cluster as repetitive sensory-motor behaviors and behaviors reflecting resistance to change. The C58 mouse strain is a promising model for these behaviors as it emits high rates of aberrant repetitive sensory-motor behaviors. The purpose of the present study was to extend characterization of the C58 model to resistance to change. This was done by comparing C58 to C57BL/6 mice on a reversal learning task under either a 100% or 80%/20% probabilistic reinforcement schedule. In addition, the effect of environmental enrichment on performance of this task was assessed as this rearing condition markedly reduces repetitive sensory-motor behavior in C58 mice. Little difference was observed between C58 and control mice under a 100% schedule of reinforcement. The 80%/20% probabilistic schedule of reinforcement generated substantial strain differences, however. Importantly, no strain difference was observed in acquisition, but C58 mice were markedly impaired in their ability to reverse their pattern of responding from the previously high density reinforcement side. Environmental enrichment did not impact acquisition under the probabilistic reinforcement schedule, but enriched C58 mice performed significantly better than standard housed C58 mice in reversal learning. Thus, C58 mice exhibit behaviors that reflect both repetitive sensory motor behaviors as well as behavior that reflects resistance to change. Moreover, both clusters of repetitive behavior were attenuated by environmental enrichment. Such findings, along with the reported social deficits in C58 mice, increase the translational value of this mouse model to autism.
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Environmental enrichment attenuates behavioral abnormalities in valproic acid-exposed autism model mice
Source:Behavioural Brain Research, Volume 333
Author(s): Hiroshi Yamaguchi, Yuta Hara, Yukio Ago, Erika Takano, Shigeru Hasebe, Takanobu Nakazawa, Hitoshi Hashimoto, Toshio Matsuda, Kazuhiro Takuma
We recently demonstrated that prenatal exposure to valproic acid (VPA) at embryonic day 12.5 causes autism spectrum disorder (ASD)-like phenotypes such as hypolocomotion, anxiety-like behavior, social deficits and cognitive impairment in mice and that it decreases dendritic spine density in the hippocampal CA1 region. Previous studies show that some abnormal behaviors are improved by environmental enrichment in ASD rodent models, but it is not known whether environmental enrichment improves cognitive impairment. In the present study, we examined the effects of early environmental enrichment on behavioral abnormalities and neuromorphological changes in prenatal VPA-treated mice. We also examined the role of dendritic spine formation and synaptic protein expression in the hippocampus. Mice were housed for 4 weeks from 4 weeks of age under either a standard or enriched environment. Enriched housing was found to increase hippocampal brain-derived neurotrophic factor mRNA levels in both control and VPA-exposed mice. Furthermore, in VPA-treated mice, the environmental enrichment improved anxiety-like behavior, social deficits and cognitive impairment, but not hypolocomotion. Prenatal VPA treatment caused loss of dendritic spines in the hippocampal CA1 region and decreases in mRNA levels of postsynaptic density protein-95 and SH3 and multiple ankyrin repeat domains 2 in the hippocampus. These hippocampal changes were improved by the enriched housing. These findings suggest that the environmental enrichment improved most ASD-like behaviors including cognitive impairment in the VPA-treated mice by enhancing dendritic spine function.
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MMP-2: A modulator of neuronal precursor activity and cognitive and motor behaviors
Source:Behavioural Brain Research, Volume 333
Author(s): Qi Li, Michael Michaud, Ravi Shankar, Sandra Canosa, Michael Schwartz, Joseph A. Madri
Matrix Metalloproteinase2, (MMP2, gelatinase A) is a zinc-containing enzyme with a broad substrate specificity including components of the extracellular matrix, cell surface molecules and a wide range bioactive molecules. MMP2 is known to play important roles in a variety of signaling pathways and processes in a wide range of cell types and tissues.In this report we elucidate the effects of the absence of MMP2 in Neural Precursor Cells (NPC) derived from C57BL/6 MMP2 KO mice and in primary and secondary neurosphere formation. We observed smaller neurosphere numbers and sizes, decreased NPC numbers, PCNA expression, DNA and Akt activation in MMP2 KO NPC compared to WT NPC. We also found decreased neurosphere formation and NPC migration outward from adherent neurospheres, decreased CXCR4 and nestin expression and increased GFAP and neuro-filament expression in MMP2 KO NPC compared to Wt NPC. MMP2 KO mice were found to exhibit increased anxiety manifested in open field activity assays compared to Wt mice. MMP2 KO mice also exhibited differences in motor activities manifested by decreased balance and endurance during Rota-rod testing.These studies illustrate an important role of MMP2 in cognitive and motor behaviors and confirm its importance in NPC activities crucial to brain development, growth and response to and recovery from injury.
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Disruption of GluA2 phosphorylation potentiates stress responsivity
Source:Behavioural Brain Research, Volume 333
Author(s): Alexandra S. Ellis, Anne Q. Fosnocht, Kelsey E. Lucerne, Lisa A. Briand
Cocaine addiction is characterized by persistent craving and addicts frequently relapse even after long periods of abstinence. Exposure to stress can precipitate relapse in humans and rodents. Stress and drug use can lead to common alterations in synaptic plasticity and these commonalities may contribute to the ability of stress to elicit relapse. These common changes in synaptic plasticity are mediated, in part, by alterations in the trafficking and stabilization of AMPA receptors. Exposure to both cocaine and stress can lead to alterations in protein kinase C–mediated phosphorylation of GluA2 AMPA subunits and thus alter the trafficking of GluA2-containing AMPARs. However, it is not clear what role AMPAR trafficking plays in the interactions between stress and cocaine. The current study utilized a mouse with a point mutation within the GluA2 subunit c-terminus resulting in a disruption of PKC-mediated GluA2 phosphorylation to examine stress responsivity. Although no differences were seen in the response to a forced swim stress in naïve mice, GluA2 K882A knock-in mice exhibited an increased stress response following cocaine self-administration. Furthermore, we demonstrated that disrupting GluA2 phosphorylation increases vulnerability to stress-induced reinstatement of both cocaine seeking and cocaine-conditioned reward. Finally, GluA2 K882A knock-in mice exhibit an increased vulnerability to social defeat as indicated by increased social avoidance. Taken together these results indicate that disrupting GluA2 phosphorylation leads to increased responsivity to acute stress following cocaine exposure and increased vulnerability to chronic stress. These results highlight the GluA2 phosphorylation site as a novel target for the stress-related disorders.
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Dopamine transporter (DAT1/SLC6A3) polymorphism and the association between being born small for gestational age and symptoms of ADHD
Source:Behavioural Brain Research, Volume 333
Author(s): K.E. Waldie, C.M. Cornforth, R.E. Webb, J.M.D. Thompson, R. Murphy, D. Moreau, R. Slykerman, A.R. Morgan, L.R. Ferguson, E.A. Mitchell
Being small for gestational age (SGA) has been established as a risk factor for Attention Deficit Hyperactivity Disorder (ADHD). Likewise, several molecular genetic studies have found a link between DAT1 and ADHD. This study investigated whether SGA moderates the effect of dopamine transporter gene variants on the risk of ADHD. A total of 546 children of European descent were genotyped at age 11 for seven DAT1 SNPs (rs6347, rs11564774, rs40184, rs1042098, rs2702, rs8179029 and rs3863145). The Strengths and Difficulties Questionnaire was used to measure symptoms of ADHD at ages 3.5, 7 and 11. We found significant gene-environment interactions between birth weight and DAT1 SNPs (rs6347, rs40184, rs1042098, rs3863145) on ADHD symptoms at 3.5 years only. Results suggest that genotypic variation of DAT1 may confer a relative protective effect against ADHD in SGA individuals. This study supports the idea that being born SGA moderates the effect of the DAT1 gene on ADHD symptoms in the preschool years and may help to explain some of the heterogeneity in ADHD outcomes.
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Editorial Board
Source:Critical Reviews in Oncology/Hematology, Volume 116
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WHO position on the use of fractional doses – June 2017, addendum to vaccines and vaccination against yellow fever WHO: Position paper – June 2013
Source:Vaccine
Author(s): World Health Organization
This article presents the World Health Organization's (WHO) recommendations on the use of fractional doses of yellow fever vaccines excerpted from the "Yellow fever vaccine: WHO position on the use of fractional doses – June 2017, Addendum to Vaccines and vaccination against yellow fever WHO: Position Paper – June 2013″, published in the Weekly Epidemiological Record [1,2].This addendum to the 2013 position paper pertains specifically to use of fractional dose YF (fYF) vaccination (fractional dose yellow fever vaccination refers to administration of a reduced volume of vaccine dose, which has been reconstituted as usual per manufacturer recommendations) in the context of YF vaccine supply shortages beyond the capacity of the global stockpile. The current WHO position on the use of yellow fever (YF) vaccine is set out in the 2013 WHO position paper on vaccines and vaccination against YF and those recommendations are unchanged.Footnotes to this paper provide a number of core references including references to grading tables that assess the quality of the scientific evidence, and to the evidence-to-recommendation table. In accordance with its mandate to provide guidance to Member States on health policy matters, WHO issues a series of regularly updated position papers on vaccines and combinations of vaccines against diseases that have an international public health impact. These papers are concerned primarily with the use of vaccines in large-scale immunization programmes; they summarize essential background information on diseases and vaccines, and conclude with WHO's current position on the use of vaccines in the global context. Recommendations on the use of Yellow Fever vaccines were discussed by SAGE in October 2016; evidence presented at these meetings can be accessed at: http://ift.tt/2tSR86g.
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Hypothesis of Improved Visualization of Microstructures in the Interventricular Septum with Ultrasound and Adaptive Beamforming
Source:Ultrasound in Medicine & Biology
Author(s): Ole Marius Hoel Rindal, Svend Aakhus, Sverre Holm, Andreas Austeng
In this work, in vivo ultrasound cardiac images created with Capon's minimum variance adaptive beamformer are compared with images acquired with the conventional delay-and-sum beamformer. Specifically, we provide three views of a human heart imaged through the parasternal short-axis, the parasternal long-axis and the apical four-chamber views. The minimum variance beamformer produced images with improved lateral resolution, resulting in better resolved speckle structure and improved edges, especially on close investigation of the interventricular septum. These improvements in image quality might possibly improve the visualization of microstructures in the human heart.
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ROS-dependent activation of RhoA/Rho-kinase in pulmonary artery: Role of Src-family kinases and ARHGEF1
Publication date: September 2017
Source:Free Radical Biology and Medicine, Volume 110
Author(s): Charles E. MacKay, Yasin Shaifta, Vladimir V. Snetkov, Asvi A. Francois, Jeremy P.T. Ward, Greg A. Knock
The role of reactive oxygen species (ROS) in smooth muscle contraction is poorly understood. We hypothesised that G-protein coupled receptor (GPCR) activation and hypoxia induce Rho-kinase activity and contraction in rat intra-pulmonary artery (IPA) via stimulation of ROS production and subsequent Src-family kinase (SrcFK) activation.The T-type prostanoid receptor agonist U46619 induced ROS production in pulmonary artery smooth muscle cells (PASMC). U46619 also induced c-Src cysteine oxidation, SrcFK auto-phosphorylation, MYPT-1 and MLC20 phosphorylation and contraction in IPA, and all these responses were inhibited by antioxidants (ebselen, Tempol). Contraction and SrcFK/MYPT-1/MLC20 phosphorylations were also inhibited by combined superoxide dismutase and catalase, or by the SrcFK antagonist PP2, while contraction and MYPT-1/MLC20 phosphorylations were inhibited by the Rho guanine nucleotide exchange factor (RhoGEF) inhibitor Y16. H2O2 and the superoxide-generating quinoledione LY83583 both induced c-Src oxidation, SrcFK auto-phosphorylation and contraction in IPA. LY83583 and H2O2-induced contractions were inhibited by PP2, while LY83583-induced contraction was also inhibited by antioxidants and Y16. SrcFK auto-phosphorylation and MYPT-1/MLC20 phosphorylation was also induced by hypoxia in IPA and this was blocked by mitochondrial inhibitors rotenone and myxothiazol. In live PASMC, sub-cellular translocation of RhoA and the RhoGEF ARHGEF1 was triggered by both U46619 and LY83583 and this translocation was blocked by antioxidants and PP2. RhoA translocation was also inhibited by an ARHGEF1 siRNA. U46619 enhanced ROS-dependent co-immunoprecipitation of ARHGEF1 with c-Src.Our results demonstrate a link between GPCR-induced cytosolic ROS or hypoxia-induced mitochondrial ROS and SrcFK activity, Rho-kinase activity and contraction. ROS and SrcFK activate RhoA via ARHGEF1.
Graphical abstract
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Role of oxidative stress in cardiovascular disease outcomes following exposure to ambient air pollution
Publication date: September 2017
Source:Free Radical Biology and Medicine, Volume 110
Author(s): Frank J. Kelly, Julia C. Fussell
Exposure to ambient air pollution is associated with adverse cardiovascular outcomes. These are manifested through several, likely overlapping, pathways including at the functional level, endothelial dysfunction, atherosclerosis, pro-coagulation and alterations in autonomic nervous system balance and blood pressure. At numerous points within each of these pathways, there is potential for cellular oxidative imbalances to occur. The current review examines epidemiological, occupational and controlled exposure studies and research employing healthy and diseased animal models, isolated organs and cell cultures in assessing the importance of the pro-oxidant potential of air pollution in the development of cardiovascular disease outcomes. The collective body of data provides evidence that oxidative stress (OS) is not only central to eliciting specific cardiac endpoints, but is also implicated in modulating the risk of succumbing to cardiovascular disease, sensitivity to ischemia/reperfusion injury and the onset and progression of metabolic disease following ambient pollution exposure. To add to this large research effort conducted to date, further work is required to provide greater insight into areas such as (a) whether an oxidative imbalance triggers and/or worsens the effect and/or is representative of the consequence of disease progression, (b) OS pathways and cardiac outcomes caused by individual pollutants within air pollution mixtures, or as a consequence of inter-pollutant interactions and (c) potential protection provided by nutritional supplements and/or pharmacological agents with antioxidant properties, in susceptible populations residing in polluted urban cities.
Graphical abstract
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Regulation of inflammation by members of the formyl-peptide receptor family
Source:Journal of Autoimmunity
Author(s): Keqiang Chen, Zhiyao Bao, Wanghua Gong, Peng Tang, Teizo Yoshimura, Ji Ming Wang
Inflammation is associated with a variety of diseases. The hallmark of inflammation is leukocyte infiltration at disease sites in response to pathogen- or damage-associated chemotactic molecular patterns (PAMPs and MAMPs), which are recognized by a superfamily of seven transmembrane, Gi-protein-coupled receptors (GPCRs) on cell surface. Chemotactic GPCRs are composed of two major subfamilies: the classical GPCRs and chemokine GPCRs. Formyl-peptide receptors (FPRs) belong to the classical chemotactic GPCR subfamily with unique properties that are increasingly appreciated for their expression on diverse host cell types and the capacity to interact with a plethora of chemotactic PAMPs and MAMPs. Three FPRs have been identified in human: FPR1–FPR3, with putative corresponding mouse counterparts. FPR expression was initially described in myeloid cells but subsequently in many non-hematopoietic cells including cancer cells. Accumulating evidence demonstrates that FPRs possess multiple functions in addition to controlling inflammation, and participate in the processes of many pathophysiologic conditions. They are not only critical mediators of myeloid cell trafficking, but are also implicated in tissue repair, angiogenesis and protection against inflammation-associated tumorigenesis. A series recent discoveries have greatly expanded the scope of FPRs in host defense which uncovered the essential participation of FPRs in step-wise trafficking of myeloid cells including neutrophils and dendritic cells (DCs) in host responses to bacterial infection, tissue injury and wound healing. Also of great interest is the FPRs are exploited by malignant cancer cells for their growth, invasion and metastasis. In this article, we review the current understanding of FPRs concerning their expression in a vast array of cell types, their involvement in guiding leukocyte trafficking in pathophysiological conditions, and their capacity to promote the differentiation of immune cells, their participation in tumor-associated inflammation and cancer progression. The close association of FPRs with human diseases and cancer indicates their potential as targets for the development of therapeutics.
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Associations between subclinical hypocalcemia and postparturient diseases in dairy cows
Source:Journal of Dairy Science
Author(s): E.M. Rodríguez, A. Arís, A. Bach
Dairy cows suffer blood Ca losses as lactation begins and might be affected by hypocalcemia in its clinical (total serum Ca concentration <1.50 mM) or subclinical form (total serum Ca concentration ≤2.14 mM). Several studies have suggested that hypocalcemia is associated with different health problems of the cow but results from different studies are not consistent. The objective of this study was to assess potential associations between subclinical hypocalcemia (SCHC) and displaced abomasum, intramammary infections, metritis, retained placenta, and ketosis. Also, the associations between SCHC and milk yield and reproductive function were evaluated. After discarding cows (32) with clinical hypocalcemia, a total of 764 cows from 6 different commercial farms were enrolled in this study. Blood samples were collected at 24 to 48 h postcalving and analyzed for total Ca concentration. Odds ratios of the different afflictions potentially associated with SCHC were calculated. Seventy-eight percent of the analyzed cows incurred SCHC. The occurrence of displaced abomasum, ketosis, retained placenta, and metritis was 3.7, 5.5, 3.4, and 4.3 times more likely, respectively, in cows that had SCHC than in cows with normocalcemia. Furthermore, the risk of incurring retained placenta or metritis increased in multiparous cows as serum Ca concentrations decreased compared with that in primiparous cows. Normocalcemic cows, independent of parity, were more likely to show their first estrus sooner after calving than SCHC cows, but no correlation was found between SCHC and other reproductive parameters. Different serum Ca concentration cutoffs were identified for several postpartum afflictions (≤1.93, ≤2.05, ≤2.05, and ≤2.10 mM for ketosis, retained placenta, metritis, and displaced abomasum, respectively). In conclusion, SCHC, defined as serum Ca ≤2.14 mM, is a frequent illness affecting the majority of the dairy cows with important repercussions on health. However, if SCHC were to be used to predict postpartum disease, different serum Ca cutoff points are likely to be needed because best predictive cutoff values varied among postpartum ketosis, displaced abomasum, retained placenta, and metritis.
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Effects of feeding alfalfa stemlage or wheat straw for dietary energy dilution on nutrient intake and digestibility, growth performance, and feeding behavior of Holstein dairy heifers
Source:Journal of Dairy Science
Author(s): Huawei Su, Matt S. Akins, Nancy M. Esser, Robin Ogden, Wayne K. Coblentz, Kenneth F. Kalscheur, Ron Hatfield
Feeding high-quality forage diets may lead to excessive weight gains and over-conditioning for dairy heifers. Restriction of energy density and dry matter intake by using low-energy forages, such as straw, is a good approach for controlling this problem. Alfalfa stems contain high fiber and moderate protein content and have the potential to be used to replace straw to reduce dietary energy. The objective of this study was to compare nutrient intakes, digestibilities, growth performance, and feeding behaviors of dairy heifers offered an alfalfa silage/corn silage high energy diet (HE; 13.1% crude protein, 65.4% total digestible nutrients, 39.7% neutral detergent fiber) with 2 energy-diluted diets that replaced various proportions of the corn or alfalfa silages with either alfalfa stemlage (STM; 12.6% crude protein, 59.1% total digestible nutrients, 46.4% neutral detergent fiber) or chopped wheat straw (WS; 12.6% crude protein, 61.9% total digestible nutrients, 43.7% neutral detergent fiber). Seventy-two pregnant Holstein heifers (16.8 ± 1.3 mo) were stratified into 3 blocks (24 heifers/block) by initial body weight (light, 440 ± 18.0 kg; medium, 486 ± 18.6 kg; heavy, 534 ± 25.1 kg), with each block composed of 3 pens (8 heifers/pen), with diets assigned randomly to 1 pen within the block. Diets were offered in a 56-d feeding trial. Both dry matter intake and energy intake were decreased with the addition of low-energy forages to the diets, but no differences in dry matter intake were observed across diluted diets. Digestibility of dry matter, organic matter, neutral detergent fiber, and apparent N were greater for HE compared with diluted diets, and for WS compared with STM. Total body weight gain (74 vs. 56 kg) and average daily gain (1.32 vs. 1.00 kg/d) were greater for heifers offered HE compared with diluted diets. Feed efficiency tended to be less for heifers offered the diluted diets compared with HE (10.7 vs. 8.6 kg of feed/kg of gain). Heifers did not sort for or against particles when offered HE. However, increased sorting behavior was observed for diluted diets. Compared with ad libitum feeding dairy heifers a diet with high nutrient content forages (corn silage and alfalfa silage), use of diet diluted with alfalfa stemlage or wheat straw is an effective feeding management strategy to control total daily dry matter and energy intake by increasing gut fill, and maintain desirable body condition and growth rates, even though the diluted diets had greater sortability.
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Technical note: Nontargeted detection of adulterated plant proteins in raw milk by UPLC-quadrupole time-of-flight-mass spectrometric proteomics combined with chemometrics
Source:Journal of Dairy Science
Author(s): Weiying Lu, Jie Liu, Boyan Gao, Xiaxia Lv, Liangli (Lucy) Yu
We built and validated a chemometric model to detect possible milk adulteration with plant proteins. Specifically, we extracted proteins in raw milk, treated with tryptic digestion, and obtained peptide fingerprints by UPLC-quadrupole time-of-flight-mass spectrometry with proteomics to differentiate authentic milks from their counterparts adulterated with nonmilk proteins. This approach is able to detect soybean and pea powder-adulterated milks at as low as 1% (wt/wt). Additionally, we obtained the characteristic peptide sequences for milk authentication by principal component analysis. The prediction accuracies for milk authentication by partial least-squares-discriminant analysis were greater than 95%. These results indicated that peptide fingerprints with the chemometric analysis could be successfully applied for milk quality control.
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Effects of depleting ionic strength on 31P nuclear magnetic resonancespectra of micellar casein during membrane separation and diafiltration of skim milk
Source:Journal of Dairy Science
Author(s): Mattia Boiani, Padraig McLoughlin, Mark A.E. Auty, Richard J. FitzGerald, Phil M. Kelly
Membrane separation processes used in the concentration and isolation of micellar casein-based milk proteins from skim milk rely on extensive permeation of its soluble serum constituents, especially lactose and minerals. Whereas extensive literature exists on how these processes influence the gross composition of milk proteins, we have little understanding of the effects of such ionic depletion on the core structural unit of micellar casein [i.e., the casein phosphate nanocluster (CPN)]. The 31P nuclear magnetic resonance (NMR) is an analytical technique that is capable of identifying soluble and organic forms of phosphate in milk. Thus, our objective was to investigate changes to the 31P NMR spectra of skim milk during microfiltration (MF) and diafiltration (DF) by tracking movements in different species of phosphate. In particular, we examined the peak at 1.11 ppm corresponding to inorganic phosphate in the serum, as well as the low-intensity broad signal between 1.5 and 3.0 ppm attributed to casein-associated phosphate in the retentate. The MF concentration and DF using water caused a shift in the relevant 31P NMR peak that could be minimized if orthophosphate was added to the DF water. However, this did not resolve the simultaneous change in retentate pH and increased solubilization of micellar casein protein. The addition of calcium in combination with orthophosphate prevented micellar casein solubilization and simultaneously contributed to preservation of the CPN structure, except for overcorrection of retentate pH in the acidic direction. A more complex DF solution, involving a combination of phosphate, calcium, and citrate, succeeded in both CPN and micellar casein structure preservation while maintaining retentate pH in the region of the original milk pH. The combination of 31P NMR as an analytical technique and experimental probe during MF/DF processes provided useful insights into changes occurring to CPN while retaining the micellar state of casein.
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Transient changes in milk production efficiency and bacterial community composition resulting from near-total exchange of ruminal contents between high- and low-efficiency Holstein cows
Source:Journal of Dairy Science
Author(s): Paul J. Weimer, Madison S. Cox, Tania Vieira de Paula, Miao Lin, Mary Beth Hall, Garret Suen
The objectives of this study were to determine if milk production efficiency (MPE) is altered by near-total exchange of ruminal contents between high- (HE) and low-MPE (LE) cows and to characterize ruminal bacterial community composition (BCC) before exchange and over time postexchange. Three pairs of ruminally cannulated, third-lactation cows were selected whose MPE (energy-corrected milk per unit of dry matter intake) differed over their first 2 lactations. Approximately 95% of ruminal contents were exchanged between cows within each pair. Ruminal pH and volatile fatty acid (VFA) profiles, along with BCC (characterized by sequencing of the variable 4 region of 16S rRNA genes), were assessed just before feeding on d −8, −7, −5, −4, −1, 1, 2, 3, 7, 10, 14, 21, 28, 35, 42, and 56, relative to the exchange date. High-MPE cows had higher total ruminal VFA concentrations, higher molar percentages of propionate and valerate, and lower molar percentages of acetate and butyrate than did LE cows, and re-established these differences 1 d after contents exchange. Across all LE cows, MPE increased during 7 d postexchange but declined thereafter. Two of the 3 HE cows displayed decreases in MPE following introduction of the ruminal contents from the corresponding LE cow, but MPE increased in the third HE cow, which was determined to be an outlier. For all 6 cows, both liquid- and solids-associated BCC differed between individuals within a pair before contents exchange. Upon exchange, BCC of both phases in all 3 pairs was more similar to that of the donor inoculum than to preexchange host BCC. For 5 of 6 cows, the solids-associated community returned within 10 d to more resemble the preexchange community of that host than that of the donor community. Individual variability before the exchange was greater in liquids than in solids, as was the variability in response of bacterial communities to the exchange. Individual cows varied in their response, but generally moved toward re-establishment of their preexchange communities by 10 d after contents exchange. By contrast, ruminal pH and VFA profiles returned to preexchange levels within 1 d. Despite the small number of cows studied, the data suggest an apparent role for the ruminal bacterial community as a determinant of MPE.
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Experimental evidence for previously unclassified calcium phosphate structures in the casein micelle
Source:Journal of Dairy Science
Author(s): J.P. Hindmarsh, P. Watkinson
1H-31P Cross-polarization magic angle spinning (CP-MAS) measurements of 40-d-old Mozzarella cheese and 20 mM EDTA-treated casein micelles revealed that each sample had immobile phosphorus with the same spectral pattern, which did not match that of native casein micelles. To identify the immobile phosphorus bodies, 1H-31P CP-MAS spectra and cross-polarization kinetics measurements were undertaken on native casein micelles, EDTA-chelated casein micelles, and reference samples of β-casein and hydroxyapatite. The results showed that the immobile phosphorus bodies in the mature Mozzarella cheese had the following characteristics: they are immobile phosphoserine residues (not colloidal calcium phosphate); they are not the product of phosphoserine to colloidal calcium phosphate bridging; the phosphate is complexed to calcium; their rigidity is localized to a phosphorus site; their rigidity and bond coupling is unaffected by protein hydration; and the immobile bodies share a narrow range of bond orientations. Combining these observations, the best explanation of the immobile phosphorus bodies is that bonding structures of phosphorus-containing groups and calcium exist within the casein micelle that are not yet clearly classified in the literature. The best candidate is a calcium-bridged phosphoserine-to-phosphoserine linkage, either intra- or inter-protein.
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Influence of food deprivation on intravenous glucose tolerance test traits in Holstein Friesian heifers
Source:Journal of Dairy Science
Author(s): L. Antonio González-Grajales, Laura Pieper, Joachim Kremer, Rudolf Staufenbiel
Glucose is essential for numerous cellular functions due to its involvement in energy supply from early development to adulthood. In the lactating cow, glucose demands by the mammary gland significantly increase to support milk production when compared with other tissues. Thus, insufficient blood glucose levels might predispose dairy cows to metabolic disturbances. The intravenous glucose tolerance test (ivGTT) is a suitable tool to characterize glucose metabolism and insulin responses, but results must be reliable and repeatable. One factor influencing ivGTT is food deprivation period, which has been considered as an obligatory requirement before conduction of glucose tolerance studies in monogastric species, whereas it has been permissive in ruminants. The objective of this study was to determine the influence of 5 fasting periods (0, 12, 24, 36, and 48 h) on ivGTT traits and insulin responses in German Holstein heifers. A total of 140 tests were conducted in 28 females aged 12 to 19 mo. Blood samples were collected every 7 min within 1 h. Assessed glucose and insulin parameters included basal serum glucose and insulin concentration, maximum glucose and insulin concentration obtained between min 7 to 21, and concentrations at min 63 (last sampling) relative to glucose administration, glucose area equivalent (GA), glucose area under the curve (GAUC), insulin area equivalent (InsA), insulin area under the curve (InsAUC), and blood glucose half-life time (GHLT). Serum glucose and insulin concentration were measured according to the hexokinase colorimetric method and solid phase radio immunoassay, respectively. The generalized linear mixed model was used to test for significant differences in all glucose traits and insulin responses at different fasting periods. The model used season and weight as confounding variables. Glucose and insulin concentrations at 0, 7 to 21 (maximum concentration), and 63 min were affected by the duration of food deprivation. The GA, InsA, GAUC, InsAUC, and especially GHLT were also affected by fasting period. A positive linear relationship between GHLT and length of food deprivation was found. Significantly higher GA, GAUC, GHLT values, and glucose and insulin concentration at min 63 were obtained at increasing fasting periods. High intraclass correlation coefficient (0.48) was found for GHLT. The results demonstrate that GHLT might be used to reliably characterize an individual's glucose metabolism and the importance of standardizing food deprivation schedules when performing an ivGTT in nongestating Holstein heifers.
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Effects of replacing maize silage with lucerne silage and lucerne silage chop length on rumen function and milk fatty acid composition
Source:Journal of Dairy Science
Author(s): A.L. Thomson, D.J. Humphries, K.E. Kliem, M.T. Dittmann, C.K. Reynolds
The objective of this study was to investigate whether higher lucerne (Medicago sativa; alfalfa) silage inclusion rate and longer lucerne chop length improves rumen function through increased provision of physically effective fiber, when included in a maize and lucerne silage-based total mixed ration. Diets were formulated to contain a 50:50 forage:concentrate ratio [dry matter (DM) basis] and be isonitrogenous and contain equal levels of neutral detergent fiber (320 g/kg). The forage portion of the offered diets was composed of maize and lucerne silage DM in proportions (wt/wt) of either 25:75 (high lucerne; HL) or 75:25 (low lucerne; LL). Second-cut lucerne was harvested and conserved as silage at either a long (L) or a short (S) chop length (geometric mean particle lengths of 9.0 and 14.3 mm, respectively). These variables were combined in a 2 × 2 factorial arrangement to give 4 treatments (HLL, HLS, LLL, LLS), which were fed in a 4 × 4 Latin square design study to 4 rumen-cannulated, multiparous, Holstein dairy cows in mid lactation. Effects on DM intake, chewing behavior, rumen volatile fatty acid concentration, rumen pH, rumen and fecal particle size, milk production, and milk fatty acid profile were measured. Longer chop length increased rumination times per kilogram of DM intake (+2.8 min/kg) relative to the S chop length, with HLL diets resulting in the most rumination chews. Rumen concentrations of total volatile fatty acids, acetate, and n-valerate were higher for the HLS diet than the other 3 diets, whereas rumen propionate concentration was lowest for the HLL diet. Physically effective fiber (particles >4 mm) percentage in the rumen mat was increased when L chop length was fed regardless of lucerne inclusion rate. No effect of treatment was observed for milk yield, although milk protein concentration was increased by L for the LL diet (+1.6 g/kg) and decreased by L for the HLL diet (−1.4 g/kg). Milk fat concentrations of total cis-18:1 (+3.7 g/100 g of fatty acids) and 18:3 n-3 (+0.2 g/100 g of fatty acids) were greater with HL. In conclusion, longer lucerne silage chop length increased time spent ruminating per kilogram of DM intake, but had no effect on rumen pH in the present study. Increasing dietary lucerne inclusion rate had no effects on rumination activity or rumen pH, but decreased the ratio of n-6:n-3 polyunsaturated fatty acid concentrations in milk fat.
http://ift.tt/2szF2eC
Genetic line comparisons and genetic parameters for endoparasite infections and test-day milk production traits
Source:Journal of Dairy Science
Author(s): Katharina May, Kerstin Brügemann, Tong Yin, Carsten Scheper, Christina Strube, Sven König
Keeping dairy cows in grassland systems relies on detailed analyses of genetic resistance against endoparasite infections, including between- and within-breed genetic evaluations. The objectives of this study were (1) to compare different Black and White dairy cattle selection lines for endoparasite infections and (2) the estimation of genetic (co)variance components for endoparasite and test-day milk production traits within the Black and White cattle population. A total of 2,006 fecal samples were taken during 2 farm visits in summer and autumn 2015 from 1,166 cows kept in 17 small- and medium-scale organic and conventional German grassland farms. Fecal egg counts were determined for gastrointestinal nematodes (FEC-GIN) and flukes (FEC-FLU), and fecal larvae counts for the bovine lungworm Dictyocaulus viviparus (FLC-DV). The lowest values for gastrointestinal nematode infections were identified for genetic lines adopted to pasture-based production systems, especially selection lines from New Zealand. Heritabilities were low for FEC-GIN (0.05–0.06 ± 0.04) and FLC-DV (0.05 ± 0.04), but moderate for FEC-FLU (0.33 ± 0.06). Almost identical heritabilities were estimated for different endoparasite trait transformations (log-transformation, square root). The genetic correlation between FEC-GIN and FLC-DV was 1.00 ± 0.60, slightly negative between FEC-GIN and FEC-FLU (−0.10 ± 0.27), and close to zero between FLC-DV and FEC-FLU (0.03 ± 0.30). Random regression test-day models on a continuous time scale [days in milk (DIM)] were applied to estimate genetic relationships between endoparasite and longitudinal test-day production traits. Genetic correlations were negative between FEC-GIN and milk yield (MY) until DIM 85, and between FEC-FLU and MY until DIM 215. Genetic correlations between FLC-DV and MY were negative throughout lactation, indicating improved disease resistance for high-productivity cows. Genetic relationships between FEC-GIN and FEC-FLU with milk protein content were negative for all DIM. Apart from the very early and very late lactation stage, genetic correlations between FEC-GIN and milk fat content were negative, whereas they were positive for FEC-FLU. Genetic correlations between FEC-GIN and somatic cell score were positive, indicating similar genetic mechanisms for susceptibility to udder and endoparasite infections. The moderate heritabilities for FEC-FLU suggest inclusion of FEC-FLU into overall organic dairy cattle breeding goals to achieve long-term selection response for disease resistance.
http://ift.tt/2tSIWmr
The relationship between Norwegian Red heifer growth and their first-lactation test-day milk yield: A field study
Source:Journal of Dairy Science
Author(s): K.S. Storli, G. Klemetsdal, H. Volden, R. Salte
Today's Norwegian Red (NR) is markedly different from the one that existed 25 yr ago due to the continuous genetic improvement of economically important traits. Still, current national recommendations on replacement heifer rearing largely are based on results from Danish studies from the late 1980s to the mid 1990s. The objectives of the present study were to gain information on (1) growth and growth profiles of modern NR replacement heifers in commercial dairy herds and (2) how growth during the rearing period affects the heifers' milk yield during their first lactation. To this end, we conducted a field study on 5 high-producing and 5 low-producing commercial dairy farms from each of 3 geographical regions in Norway. On these 30 farms, we combined repeated onsite registrations of growth on all available females from newborn to calving with registrations deriving from the Norwegian Dairy Herd Recording System. Each herd was visited 6 to 8 times over a period of 2 yr. At each visit, heart girth circumference on all available young females was measured. Registrations were made on a total of 3,110 heifers. After imposing restrictions on the data, growth parameters were estimated based on information from 536 animals, whereas 350 of these animals had the required information needed to estimate the relationship between growth and test-day milk yield. Our findings pointed toward an optimal ADG of 830 g/d from 10 to 15 mo of age that would optimize first-lactation yield of heifers in an average Norwegian dairy herd. The optimum will likely increase from selection over time. Utilizing simple proportionality, the ADG between 5 and 10 mo of age ideally should be 879 g/d, taking into account the fact that animal growth rate is higher at low ages and that a high prepubertal growth rate had no negative effect on first-lactation yield. When such a rearing practice is used to meet the requirements of today's genetically improved NR heifer, heifers can both optimize production in their first lactation and enter the milking herd earlier than the current average age of 24.8 mo.
http://ift.tt/2sznyPq
Bovine miR-146a regulates inflammatory cytokines of bovine mammary epithelial cells via targeting the TRAF6 gene
Source:Journal of Dairy Science
Author(s): Xing-Ping Wang, Zhuo-Ma Luoreng, Lin-Sen Zan, Feng Li, Na Li
It has been reported previously that bovine miR-146a (bta-miR-146a) is significantly differentially expressed in mammary glands infected with mastitis, compared with healthy udders. This suggests that bta-miR-146a plays an important role in the regulation of mammary inflammation. However, the specifics of this function have yet to be elucidated. Bovine mammary epithelial cells (bMEC) represent the first line of defense against pathogens and have important roles in initiating and regulating inflammatory responses and innate immunity during infection. In this study, a double luciferase reporter assay was used to confirm that bta-miR-146a directly targets the 3′ UTR of the tumor-necrosis factor receptor-associated factor 6 (TRAF6) gene. To elucidate the role of bta-miR-146a in innate immune responses, either a mimic or inhibitor of bta-miR-146a was transfected into bMEC stimulated with lipopolysaccharide, which activates the innate immune response through the toll-like receptor (TLR) 4/nuclear factor (NF)-κB signaling pathway. Forty-eight hours posttransfection, quantitative real-time PCR and Western blots were used to detect the expressions of the related genes and proteins, respectively. An ELISA was used to measure the quantity of inflammatory factors in culture supernatants. The results showed that bta-miR-146a significantly inhibits both mRNA and protein expression levels of bovine TRAF6, and ultimately suppresses downstream expression of NF-κB mRNA and protein. As a result, production of NF-κB-dependent inflammatory mediators such as tumor necrosis factor α, IL-6, and IL-8 are suppressed following lipopolysaccharide stimulation of bMEC. Thus, we concluded that bta-miR-146a acts as a negative feedback regulator of bovine inflammation and innate immunity through downregulation of the TLR4/TRAF6/NF-κB pathway. This study presents a potential regulatory mechanism of bta-miR-146a on immune responses in bovine mammary infection and may provide a potential therapeutic target for mastitis.
http://ift.tt/2tSLdOR
Flow cytometric assessment of myeloperoxidase in bovine blood neutrophils and monocytes
Source:Journal of Dairy Science
Author(s): Elke Depreester, Evelyne Meyer, Kristel Demeyere, Mieke Van Eetvelde, Miel Hostens, Geert Opsomer
Myeloperoxidase (MPO) is a lysosomal peroxidase enzyme mainly stored in the azurophilic granules of neutrophils playing an important role in innate immunity for first-line protection against microorganisms in many species including cattle. As such, determination of MPO has become of great interest for the diagnosis of infectious and inflammatory diseases in multiple species such as humans. In cattle, MPO determination is rarely done because methods to assess MPO in this species are limited: functional assays have been described earlier, but so far, the quantification of MPO at the single cell level has not been done yet. In the present paper, an innovative flow cytometric method to assess MPO in blood leukocytes of dairy cattle is described. A commercial anti-bovine MPO was used following density gradient separation to isolate polymorphonuclear (PMN) and mononuclear (MN) leukocytes from blood. Identification of PMN and MN, subdivided in monocytes and lymphocytes, was based on the expression of the surface markers CH138A and CD172A. The optimized protocol was subsequently evaluated on blood samples of 17 Holstein Friesian heifers. Myeloperoxidase expression was measured flow cytometrically and visualized by fluorescence microscopic imaging of sorted PMN and MN populations. We suggest this innovative method to be useful in the field for early detection of cows at higher risk for inflammatory diseases such as mastitis and metritis during the transition period.
http://ift.tt/2tSEVyH
Genetically modified Lactococcus lactis producing a green fluorescence protein–bovine lactoferrin fusion protein suppresses proinflammatory cytokine expression in lipopolysaccharide-stimulated RAW 264.7 cells
Source:Journal of Dairy Science
Author(s): S. Shigemori, F. Namai, Y. Yamamoto, S. Nigar, T. Sato, T. Ogita, T. Shimosato
Lactoferrin (LF), an iron-binding glycoprotein distributed widely in the biological fluids of mammals, is believed to play an important role in host defenses against infection. Previous studies in animal models and humans demonstrated that combined administration of LF and probiotic lactic acid bacteria (LAB) can prevent sepsis. In this study, we genetically engineered a probiotic LAB strain, Lactococcus lactis, to produce recombinant bovine LF based on the green fluorescence protein (GFP)-fused expression system. Western blotting confirmed that the genetically modified L. lactis strain (designated NZ-GFP-bLF) produced a protein corresponding to a fusion of GFP and bLF in the presence of nisin, an inducer of target gene expression. The protein synthesized by NZ-GFP-bLF was fluorescent and thus we monitored the time-dependent change in the production level of the recombinant protein using fluorometric analysis. The utility of NZ-GFP-bLF in preventing sepsis was determined by investigating its anti-inflammatory property in lipopolysaccharide (LPS)-stimulated mouse macrophage RAW 264.7 cells. Pretreatment of RAW 264.7 cells with NZ-GFP-bLF significantly attenuated the LPS-induced mRNA expression and protein production of 3 proinflammatory cytokines (IL-1α, IL-6, and tumor necrosis factor-α) compared with pretreatment with a vector control strain of L. lactis. Our results suggest that NZ-GFP-bLF holds promise for the development of a new prophylaxis for sepsis.
http://ift.tt/2szz9hh
Phase II Trial of Sorafenib in Combination with Capecitabine in Patients with Hepatocellular Carcinoma: INST 08‐20
AbstractLessons Learned. There continues to be a lack of systemic options for advanced hepatocellular carcinoma (HCC); sorafenib and, very recently, regorafenib are the only approved options. There exists a potential to combine sorafenib with chemotherapeutic agents shown to be active in HCC, such as capecitabine, safely.Good tumor response was observed, with objective improvement in a few patients seldom seen by single agent sorafenib; however, because of the limited number of patients, meaningful conclusions on survival cannot be drawn.Background.Sorafenib is the currently approved first‐line treatment for hepatocellular carcinoma (HCC). Capecitabine has antitumor activity in hepatobiliary cancers. The combination of the two, if tolerated, could possibly improve antitumor response, and survival.Methods.Patients with advanced HCC ineligible for locoregional therapy, Eastern Cooperative Oncology Group performance status of ≤2, Child‐Pugh class A or B‐7 cirrhosis, hemoglobin ≥8.5 g/dL, platelets ≥50,000/μL, absolute neutrophil count (ANC) ≥1500 cells/μL, and serum creatinine of ≤2.0 mg/dL were recruited. All subjects received a combination of sorafenib and capecitabine, on a 14‐day 7‐days on 7‐days off schedule. The primary end point was safety and secondary end points were overall survival (OS) and disease control rate.Results.A total of 15 out of 47 patients met inclusion criteria. Median age was 64 years (56–79) and 77% were male. With a median follow‐up of 12 months, median OS was 12.7 months (95% confidence interval [CI], 8.5–23.4). Disease control rate was 77% (complete response 8%, partial response 8%, and stable disease 61%). Common adverse events were as follows: (a) thrombocytopenia (64%); (b) anemia (14%); (c) hypophosphatemia (21%); (d) hypomagnesemia (14%); (e) hyperbilirubinemia (21%); (f) increased aspartate transaminase (AST) (14%); (g) hand‐foot syndrome (21%); and (h) deep vein thrombosis (21%).Conclusion.At tolerable doses, the combination of sorafenib and capecitabine seems an active and safe palliative treatment for HCC in class A and B‐7 patients with cirrhosis. The small sample size does not allow comparison with single‐agent sorafenib.
http://ift.tt/2uBcEKf
Antiemetic Corticosteroid Rotation from Dexamethasone to Methylprednisolone to Prevent Dexamethasone‐Induced Hiccup in Cancer Patients Treated with Chemotherapy: A Randomized, Single‐Blind, Crossover Phase III Trial
AbstractBackground.To assess whether the rotation of dexamethasone to methylprednisolone decreases the intensity of dexamethasone‐induced hiccup (DIH) in cancer patients treated with chemotherapy.Materials and Methods.Adult patients who experienced DIH within 3 days after the administration of dexamethasone as an antiemetic were screened. Eligible patients were randomly assigned to receive dexamethasone (n = 33) or methylprednisolone (n = 32) as an antiemetic (randomization phase). In the next cycle of chemotherapy, the dexamethasone group received methylprednisolone and vice versa in the methylprednisolone group (crossover phase). The primary endpoint was the difference in hiccup intensity as measured using the numeric rating scale (NRS) between two groups.Results.No female patients were enrolled, although the study did not exclude them. At the randomization phase, hiccup frequency was 28/33 (84.8%) in the dexamethasone group versus 20/32 (62.5%) in the methylprednisolone group (p = .04). Intensity of hiccup was significantly higher in the dexamethasone group than that in the methylprednisolone group (mean NRS, 3.5 vs. 1.4, p < .001). At the crossover phase, hiccup intensity was further decreased after the rotation of dexamethasone to methylprednisolone in the dexamethasone group (mean NRS, 3.5 to 0.9, p < .001), while it was increased by rotating methylprednisolone to dexamethasone in the methylprednisolone group (mean NRS, 1.4 to 3.3, p = .025). There were no differences in emesis intensity between the two groups at either the randomization or crossover phases. Clinicaltrials.gov identifier: NCT01974024.Conclusion.Dexamethasone‐induced hiccup is a male‐predominant phenomenon that can be ameliorated by rotating dexamethasone to methylprednisolone without compromising the antiemetic efficacy.Implications for Practice.In this randomized, multicenter, phase III trial, hiccup intensity was significantly lower when the antiemetic corticosteroid was rotated from dexamethasone to methylprednisolone without a change in emesis intensity than that when dexamethasone was maintained. At the crossover phase, hiccup intensity was increased again if dexamethasone was readministered instead of methylprednisolone. The present study demonstrated that dexamethasone‐induced hiccup can be improved by rotating from dexamethasone to methylprednisolone without compromising its antiemetic efficacy.
http://ift.tt/2u274nm
A Phase II Trial Evaluating the Safety of Rapid Infusion of Ofatumumab in Patients with Previously Treated Chronic Lymphocytic Leukemia
AbstractLessons Learned. Ofatumumab infusion reactions can be diminished by escalating the dose rate in individual patients in sequential infusions.Background.Ofatumumab (OFA) is a fully humanized, anti‐CD20 antibody approved for use in chronic lymphocytic leukemia (CLL). The recommended administration requires long infusion times. We evaluated an accelerated infusion regimen of 2 hours.Methods.The first dose of OFA (300 mg) was given on week 1 day 1 starting at 3.6 mg/hour and doubling every 30 minutes until a rate of 240 mg/hour was reached. If tolerated, the second dose (1,000 mg) was given on week 1 day 3 starting at 50 mg/hour and doubling every 30 minutes until a rate of 800 mg/hour was reached. If tolerated, the third dose (2,000 mg) was given on week 2 day 1 at 800 mg/hour over the first 30 minutes and, if tolerated, at 1,068 mg/hour over the next 90 minutes (goal infusion time: 120 minutes). Subsequent OFA infusions were administered weekly in the same manner for 8 weeks, and then monthly for 4 months.Results.Thirty‐four patients were treated. Most infusion‐related reactions occurred during the first and second infusion. Eighty‐seven percent (87%) of patients finished the third infusion within 15 minutes of the planned 2 hours and only one had an infusion reaction.Conclusion.Using this stepped‐up dosing regimen, a rapid infusion of OFA is safe and well tolerated.
http://ift.tt/2uBcEdd
Selection of effective cocrystals former for dissolution rate improvement of active pharmaceutical ingredients based on lipoaffinity index
Publication date: 30 September 2017
Source:European Journal of Pharmaceutical Sciences, Volume 107
Author(s): Piotr Cysewski, Maciej Przybyłek
New theoretical screening procedure was proposed for appropriate selection of potential cocrystal formers possessing the ability of enhancing dissolution rates of drugs. The procedure relies on the training set comprising 102 positive and 17 negative cases of cocrystals found in the literature. Despite the fact that the only available data were of qualitative character, performed statistical analysis using binary classification allowed to formulate quantitative criterions. Among considered 3679 molecular descriptors the relative value of lipoaffinity index, expressed as the difference between values calculated for active compound and excipient, has been found as the most appropriate measure suited for discrimination of positive and negative cases. Assuming 5% precision, the applied classification criterion led to inclusion of 70% positive cases in the final prediction. Since lipoaffinity index is a molecular descriptor computed using only 2D information about a chemical structure, its estimation is straightforward and computationally inexpensive. The inclusion of an additional criterion quantifying the cocrystallization probability leads to the following conjunction criterions Hmix<−0.18 and ΔLA>3.61, allowing for identification of dissolution rate enhancers. The screening procedure was applied for finding the most promising coformers of such drugs as Iloperidone, Ritonavir, Carbamazepine and Enthenzamide.
Graphical abstract
http://ift.tt/2sX7UwD
Design, synthesis, and biological evaluation of 2,4-dihydropyrano[2,3-c]pyrazole derivatives as autotaxin inhibitors
Publication date: 30 September 2017
Source:European Journal of Pharmaceutical Sciences, Volume 107
Author(s): Tatu Pantsar, Prosanta Singha, Tapio J. Nevalainen, Igor Koshevoy, Jukka Leppänen, Antti Poso, Juha M.A. Niskanen, Sanna Pasonen-Seppänen, Juha R. Savinainen, Tuomo Laitinen, Jarmo T. Laitinen
Inhibition of Autotaxin (ATX) is a potential treatment strategy for several diseases, including tumors with elevated ATX-lysophosphatidic acid (LPA) signaling. Combining structure-based virtual screening together with hen egg-white Autotaxin (ewATX) activity assays enabled the discovery of novel small-molecule ATX inhibitors with a 2,4-dihydropyrano[2,3-c]pyrazole scaffold. These compounds are suggested to bind to the lipophilic pocket, leaving the active site unrestrained. Our most potent compound, (S)-6-amino-4-(3,4-dichlorophenyl)-3-(4-[(4-fluorobenzyl)oxy]phenyl)-2,4-dihydropyrano[2,3-c]pyrazole-5-carbonitrile [(S)-25], inhibited human ATX (hATX) with an IC50-value of 134nM. It also blocked ATX-evoked but not LPA-mediated A2058 melanoma cell migration. Noteworthy, molecular modeling correctly predicted the biologically active enantiomer of 2,4-dihydropyrano[2,3-c]pyrazoles, as verified by compound crystallization and activity assays. Our study established the ewATX activity assay as a valid and affordable tool in ATX inhibitor discovery. Overall, our study offers novel insights and approaches into design of novel ATX inhibitors targeting the hydrophobic pocket instead of the active site.
Graphical abstract
http://ift.tt/2sXkqMp
Tritium forms discrimination in ryegrass under constant tritium exposure: From seed germination to seedling autotrophy
Source:Journal of Environmental Radioactivity, Volume 177
Author(s): H. Renard, D. Maro, S. Le Dizès, A. Escobar-Gutiérrez, C. Voiseux, L. Solier, D. Hébert, M. Rozet, C. Cossonnet, R. Barillot
Uncertainties remain regarding the fate of atmospheric tritium after it has been assimilated in grasslands (ryegrass) in the form of TFWT (Tissue Free Water Tritium) or OBT (Organically Bound Tritium). One such uncertainty relates to the tritium forms discrimination during transfer from TFWT to OBT resulting from photosynthesis (OBTphoto), corresponding to the OBTphoto/TFWT ratio. In this study, the OBT/TFWT ratio is determined by experiments in the laboratory using a ryegrass model and hydroponic cultures, with constant activity of tritium in the form of tritiated water (denoted as HTO) in the "water" compartment (liquid HTO) and "air" compartment (HTO vapour in the air). The OBTphoto/TFWT ratio and the exchangeable OBT fraction are measured for three parts of the plant: the leaf, seed and root. Plant growth is modelled using dehydrated biomass measurements taken over time in the laboratory and integrating physiological functions of the plant during the first ten days after germination. The results suggest that there is no measurable discrimination of tritium in the plant organic matter produced by photosynthesis.
http://ift.tt/2tWfVWQ
Influence of bisphosphonates on the adherence and metabolism of epithelial cells and gingival fibroblasts to titanium surfaces
Abstract
Objectives
To evaluate the effects of sodium alendronate (SA) and zoledronic acid (ZA), on the adhesion and metabolism of epithelial cells and gingival fibroblasts to titanium surfaces considering cell functions related to an effective mucosal barrier around the implant.
Materials and methods
Cells were seeded onto titanium discs and incubated for 24 h. Then, serum-free DMEM containing selected bisphosphonates (0, 0.5, 1, or 5 μM) was added for 24 and 48 h. Factors related to the achievement of an effective mechanical and immunological barrier—cell adhesion, viability, collagen epidermal growth factor, and immunoglobulin synthesis—were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests as well as by ANOVA and Tukey's tests, (α = 0.05).
Results
The presence of bisphosphonates culminated in lower cell adhesion to the titanium discs, particularly for SA at 5 μM (40%) and ZA at all concentrations (from 30 to 50%, according to increased concentrations). Reduced cell viability occurred after exposing these cells to ZA (40%); however, only 5 μM SA-treated cells had decreased viability (30%). Reduced synthesis of growth factors and collagen was observed when cells were reated with ZA (20 and 40%, respectively), while about 70% of IgG synthesis was enhanced.
Conclusion
Bisphosphonates negatively affected the adhesion and metabolism of oral mucosal cells, and this effect was related to the type of bisphosphonate as well as to concentration and period of treatment.
Clinical relevance
The negative effects of bisphosphonates on oral mucosal cells can hamper the formation of an effective biological seal in osseointegrated implants.
http://ift.tt/2tr3tgV
Mass-loss rates from decomposition of plant residues in spruce forests near the northern tree line subject to strong air pollution
Abstract
Mass-loss rates during the early phase of decomposition of plant residues were studied for a period of 3 years in Norway spruce forests subjected to air pollution by Cu-Ni smelters on the Kola Peninsula, northwest Russia. Litterbags were deployed in two main patches of forests at the northern tree line, between and below the crowns of spruce trees older than 100 years. The study results demonstrated the dependence of the decomposition rates on the initial concentrations of nutrients and the C/N and lignin/N ratios in plant residues. Lower rates of mass loss in forests subject to air pollution may be related to low quality of plant residues, i.e. high concentrations of heavy metals, low concentrations of nutrients, and high lignin/N and C/N ratios. The increased losses of Ca, Mg, K, and Mn from plant residues in these forests compared to the reference were, probably, related to leaching of their compounds from the residues. The relatively high rates of heavy metal accumulation in the residues were most likely related to uptake of pollutants from the atmosphere, as well as to the lower mass-loss rates. The present study results demonstrate that the forest patchiness should be taken into account in assessment and predictions of decomposition rates in Norway spruce forests. Mass-loss rates of plant residues below the crowns of old spruce trees were significantly lower than those in the patches between the crowns. This was explained by the high C/N and lignin/N ratios in the residues of evergreens which contribute significantly to litterfall below the crowns and by lower soil temperature during winter and spring below the crowns. In addition, a lower amount of precipitation reaching the forest floor below the dense, long crowns of old Norway spruce trees may result in considerably lower washing out of the organic compounds from the residues. Lower mass-loss rates below the crowns of old spruce trees may be part of the evidence that the old-growth spruce forests can continue to accumulate carbon in soil.
http://ift.tt/2sWhAaF
Interpretation of single and competitive adsorption of cadmium and zinc on activated carbon using monolayer and exclusive extended monolayer models
Abstract
In this work, a modeling analysis based on experimental tests of cadmium/zinc adsorption, in both single-compound and binary systems, was carried out. All the experimental tests were conducted at constant pH (around neutrality) and temperature (20 °C). The experimental results showed that the zinc adsorption capacity was higher than that of cadmium and it does not depend on cadmium presence in binary system. Conversely, cadmium adsorption is affected by zinc presence. In order to provide good understanding of the adsorption process, two statistical physics models were proposed. A monolayer and exclusive extended monolayer models were applied to interpret the single-compound and binary adsorption isotherms of zinc and cadmium on activated carbon. Based on these models, the modeling analysis demonstrated that zinc is dominant in solution and more favorably adsorbed on activated carbon surface. For instance, in single-compound systems, the number of ions bound per each receptor site was n (Zn2+) = 2.12 > n (Cd2+) = 0.98. Thus, the receptor sites of activated carbon are more selective for Zn2+ than for Cd2+. Moreover, the determination of adsorption energy through the adopted models confirmed that zinc is more favored for adsorption in single-compound system (adsorption energies equal to 12.12 and 7.12 kJ/mol for Zn and Cd, respectively) and its adsorption energy does not depend on the cadmium presence in binary system. Finally, the adsorption energy values suggested that single-compound and binary adsorption of zinc and cadmium is a physisorption.
http://ift.tt/2tr0x3O
Distribution and evolution of sterols and aliphatic hydrocarbons in dated marine sediment cores from the Cabo Frio upwelling region, SW Atlantic, Brazil
Abstract
We report the distribution of selected lipid biomarkers specifically sterols and aliphatic hydrocarbons in sediment cores from Cabo Frio, SW Atlantic continental shelf, Brazil, corresponding approximately to the last 700 years. In the Cabo Frio region, a costal upwelling occurs as a quasi-seasonal phenomenon characterized by nutrient-rich bottom waters that intrude on the continental shelf and promote relatively high biological productivity compared to other Brazilian continental shelf areas. The results for sterols indicate the predominance of organic matter (OM) inputs related to marine organisms, mainly plankton, in all of the cores along the time scale studied. Principal component analyses show three different groups of variables, which may be associated with (i) the more effective intrusion of the nutrient-rich South Atlantic Central Water, resulting in the increase of marine lipid biomarkers such as sterols and short-chain n-alkanes; (ii) the influence of the Coastal Water with higher surface water temperature and subsequently lower primary productivity; and (iii) OM characterized by high total organic carbon and long-chain n-alkanes related to an allochthonous source. Relatively high concentrations of sterols and n-alkanes between 1450 and 1700 AD, chronologically associated with the Little Ice Age, suggest a period associated with changes in the local input of specific sources of these compounds. The concentrations of lipid biomarkers vary over core depth, but this does not suggest a notably high or low intensity of upwelling processes. It is possible that the climatic and sea surface temperature changes reported in previous studies did not affect the input of the sedimentary lipid biomarkers analyzed here.
http://ift.tt/2sWjdoI
Mechanical loading increases pro-inflammatory effects of nitrogen-containing bisphosphonate in human periodontal fibroblasts
Abstract
Objectives
There is increasing evidence that inflammation and biomechanical loading can influence the effects of bisphosphonates (BP). The aim of this study was to investigate the influence of tensile strain application combined with IL-1ß and clodronate or zoledronate on human periodontal ligament fibroblasts (HPdLF) in vitro.
Materials and methods
HPdLF were cultured with 10 nM IL-1ß and 5 μM clodronate or zoledronate for 48 h. Cells were applied to cyclic tensile strain (CTS; 3% elongation) for 12 h in vitro. Cell number was analyzed directly after CTS by MTT assay. Gene expression of receptor activator of cyclooxygenase-2 (COX-2) was investigated using real-time PCR. MMP-8, TIMP-1, and PGE2 were measured by ELISA. Statistics were performed with SPSS (ANOVA, p < 0.05).
Results
Zoledronate reduced the cell number of HPdLF (60.3 vs. 100%), which was significant when combined with IL-1ß. Combined with 3% CTS, this effect was voided and cell number increased over the level of the control cells. IL-1ß led to a 10-fold increase of COX-2 gene expression. Combined with CTS and zoledronate, this increase was enhanced to a gene expression 70-fold that of control cells with related PGE2 synthesis. Clodronate neither reduce the cell number nor enhanced the COX-2 gene expression. CTS increased MMP-8 protein synthesis. Combined with BP, this increase was voided. TIMP-1 protein synthesis was increased at all conditions under CTS.
Conclusions
Mechanical loading might activate cell metabolism and abolish BP- and inflammation-induced reduction of viability. Combination of mechanical loading, inflammation, and nitrogen-containing bisphosphonates can cause pro-inflammatory effects.
Clinical relevance
Periodontal inflammation should be treated initially before BP intake to prevent decreased cell viability of the periodontium and increased inflammation, which might be enhanced by the addition of mastication forces.
http://ift.tt/2tqJzCO
Measurement and decomposition of energy efficiency of Northeast China—based on super efficiency DEA model and Malmquist index
Abstract
Nowadays, environment problem has become the international hot issue. Experts and scholars pay more and more attention to the energy efficiency. Unlike most studies, which analyze the changes of TFEE in inter-provincial or regional cities, TFEE is calculated with the ratio of target energy value and actual energy input based on data in cities of prefecture levels, which would be more accurate. Many researches regard TFP as TFEE to do analysis from the provincial perspective. This paper is intended to calculate more reliably by super efficiency DEA, observe the changes of TFEE, and analyze its relation with TFP, and it proves that TFP is not equal to TFEE. Additionally, the internal influences of the TFEE are obtained via the Malmquist index decomposition. The external influences of the TFFE are analyzed afterward based on the Tobit models. Analysis results demonstrate that Heilongjiang has the highest TFEE followed by Jilin, and Liaoning has the lowest TFEE. Eventually, some policy suggestions are proposed for the influences of energy efficiency and study results.
http://ift.tt/2syJudp
Pembrolizumab, Chemotherapy, and Radiation Therapy With or Without Surgery in Treating Patients With Anaplastic Thyroid Cancer
Interventions: Drug: Docetaxel; Drug: Doxorubicin Hydrochloride; Radiation: Intensity-Modulated Radiation Therapy; Other: Laboratory Biomarker Analysis; Biological: Pembrolizumab; Procedure: Therapeutic Conventional Surgery
Sponsors: Mayo Clinic; National Cancer Institute (NCI)
Not yet recruiting - verified July 2017
http://ift.tt/2uTDEnC
Effects of Voluntary Neck Extension on Cerebral Blood Flow, in Breath-hold Divers Ending an Apnoea of Two Minutes or More.
Intervention: Other: apnea performing
Sponsor: Nantes University Hospital
Not yet recruiting - verified July 2017
http://ift.tt/2tVxvu7
Use of Intra-Operative Shear Wave Ultrasound Vibrometry for Characterization of Esophageal Malignant Tumors
Intervention: Device: General Electric LOGIQ E9 ultrasound system
Sponsor: Mayo Clinic
Recruiting - verified July 2017
http://ift.tt/2tVF0RJ
NCI-MATCH: Targeted Therapy Directed by Genetic Testing in Treating Patients With Advanced Refractory Solid Tumors, Lymphomas, or Multiple Myeloma
Interventions: Drug: Afatinib; Drug: Akt inhibitor AZD5363; Drug: Binimetinib; Drug: Crizotinib; Other: Cytology Specimen Collection Procedure; Drug: Dabrafenib; Drug: Dasatinib; Drug: Defactinib; Drug: FGFR Inhibitor AZD4547; Other: Laboratory Biomarker Analysis; Biological: Nivolumab; Drug: Osimertinib; Drug: Palbociclib; Biological: Pertuzumab; Drug: PI3K-beta Inhibitor GSK2636771; Drug: Sapanisertib; Drug: Sunitinib Malate; Drug: Taselisib; Drug: Trametinib; Biological: Trastuzumab Emtansine; Drug: Trk Inhibitor LOXO-101; Drug: Vismodegib; Drug: WEE1 Inhibitor AZD1775
Sponsor: National Cancer Institute (NCI)
Recruiting - verified July 2017
http://ift.tt/2u0CpXt
Cells in 3D-reconstitutued eccrine sweat gland cell spheroids differentiate into gross cystic disease fluid protein 15-expressing dark secretory cells and carbonic anhydrase II-expressing clear secretory cells
Source:Acta Histochemica
Author(s): Haihong Li, Liyun Chen, Mingjun Zhang, Bingna Zhang
Secretory coils of eccrine sweat glands are composed of myoepithelial cells, dark secretory cells and clear secretory cells. The two types of cells play important roles in sweat secretion. In our previous study, we demonstrated that the 3D-reconstituted eccrine sweat gland cell spheroids differentiate into secretory coil-like structures. However, whether the secretory coil-like structures further differentiate into dark secretory cells and clear secretory cells were is still unknown. In this study, we detected the differentiation of clear and dark secretory cells in the 3D-reconstituted eccrine sweat gland cell spheroids using the dark secretory cell-specific marker, GCDFP-15, and clear secretory cell-specific marker, CAII by immunofluorescence staining. Results showed that there were both GCDFP-15- and CAII-expressing cells in 12-week-old 3D spheroids, similar to native eccrine sweat glands, indicating that the spheroids possess a cellular structure capable of sweat secretion. We conclude that the 12-week 3D spheroids may have secretory capability.
http://ift.tt/2sVuQMR
OMFS: Notation confusion
British Dental Journal 223, 3 (2017). doi:10.1038/sj.bdj.2017.559
Author: M. V. B. Nelson
http://ift.tt/2tRbRaH
Orienting digital radiographs
British Dental Journal 223, 3 (2017). doi:10.1038/sj.bdj.2017.560
Authors: K. Ali & P. Ward
http://ift.tt/2tQQfLw
Are people with an orofacial cleft at a higher risk of dental caries? A systematic review and meta-analysis
British Dental Journal 223, 37 (2017). doi:10.1038/sj.bdj.2017.581
Authors: V. Worth, R. Perry, T. Ireland, A. K. Wills, J. Sandy & A. Ness
http://ift.tt/2tRbPQ7
Burden to A&E
British Dental Journal 223, 3 (2017). doi:10.1038/sj.bdj.2017.561
Authors: E. Bowden & H. Cashman
http://ift.tt/2tRBJDc
Dental radiography: Cherry picking evidence
British Dental Journal 223, 4 (2017). doi:10.1038/sj.bdj.2017.562
Authors: M. Armstrong, R. Ladwa & A. Lockyer
http://ift.tt/2tQSs9D
Future gazing
British Dental Journal 223, 1 (2017). doi:10.1038/sj.bdj.2017.558
Author: Stephen Hancocks
http://ift.tt/2tR7tbz
Oral health: Praise for National Smile Month
British Dental Journal 223, 4 (2017). doi:10.1038/sj.bdj.2017.563
Author: C. Jimenez
http://ift.tt/2tRdXYe
How did we do? An investigation into the suitability of patient questionnaires (PREMs and PROMs) in three primary care oral surgery practices
British Dental Journal 223, 27 (2017). doi:10.1038/sj.bdj.2017.582
Authors: G. Gerrard, R. Jones & R. J. Hierons
http://ift.tt/2tR7ufD
Career satisfaction and work-life balance of specialist orthodontists within the UK/ROI
British Dental Journal 223, 53 (2017). doi:10.1038/sj.bdj.2017.585
Authors: S. M. Al-Junaid, S. J. Hodges, A. Petrie & S. J. Cunningham
http://ift.tt/2tRbJrJ
Perspectives: 'Step by step we build up a rapport'
British Dental Journal 223, 6 (2017). doi:10.1038/sj.bdj.2017.566
Author: Kate Quinlan
Is it realistic to see all patients by the age of one, as advocated by the British Society of Paediatric Dentistry?
http://ift.tt/2tRdUf0
Reply to Nevens et al.
Source:Radiotherapy and Oncology
Author(s): Robert J. Amdur, Roi Dagan, William M. Mendenhall
http://ift.tt/2uSH3Dk
“Radio-oncomics”
Abstract
Introduction
Radiomics, a recently introduced concept, describes quantitative computerized algorithm-based feature extraction from imaging data including computer tomography (CT), magnetic resonance imaging (MRT), or positron-emission tomography (PET) images. For radiation oncology it offers the potential to significantly influence clinical decision-making and thus therapy planning and follow-up workflow.
Methods
After image acquisition, image preprocessing, and defining regions of interest by structure segmentation, algorithms are applied to calculate shape, intensity, texture, and multiscale filter features. By combining multiple features and correlating them with clinical outcome, prognostic models can be created.
Results
Retrospective studies have proposed radiomics classifiers predicting, e. g., overall survival, radiation treatment response, distant metastases, or radiation-related toxicity. Besides, radiomics features can be correlated with genomic information ("radiogenomics") and could be used for tumor characterization.
Discussion
Distinct patterns based on data-based as well as genomics-based features will influence radiation oncology in the future. Individualized treatments in terms of dose level adaption and target volume definition, as well as other outcome-related parameters will depend on radiomics and radiogenomics. By integration of various datasets, the prognostic power can be increased making radiomics a valuable part of future precision medicine approaches.
Conclusion
This perspective demonstrates the evidence for the radiomics concept in radiation oncology. The necessity of further studies to integrate radiomics classifiers into clinical decision-making and the radiation therapy workflow is emphasized.
http://ift.tt/2tV18LT
Spatial and temporal expression of Sox9 during murine incisor development
Abstract
Mouse incisors are capable of continuously growing due to the renewal of dental epithelium stem cells and mesenchymal stem cells residing at the proximal ends. The transcription factor Sox9 plays important roles in maintaining the stem cells of hair follicles, retinal progenitor cells and neural crest stem cells. Whether Sox9 is involved during mouse incisor development is not reported yet. In this study, we examined the expression pattern of Sox9 during mouse incisor development by in situ hybridization and immunohistochemistry. Sox9 mRNA and protein showed similar expression pattern from embryonic day (E) 13.5 to postnatal (PN) day 10. At E13.5 and E14.5, Sox9 was strongly expressed in the dental epithelium. At E16.5, Sox9 started to be detected in the mesenchymal cells within the dental pulp, especially the dental pulp cells that adjacent to the labial cervical loop. Similarly with E14.5, Sox9 was strongly detected in the labial cervical loop, including the basal epithelium, the stellate reticulum and the outer enamel epithelium from E16.5 to PN10. The mesenchyme adjacent to the labial cervical loop also showed strong signal of Sox9. The spatiotemporal expression of Sox9 suggested its possible involvement during mouse incisor development.
http://ift.tt/2tV2M01
Early childhood myoclonic epilepsy: An independent genetic generalized epilepsy with myoclonic seizures as the main seizure type
Source:Clinical Neurophysiology
Author(s): Zhixian Yang, Jiao Xue, Hui Li, Ping Qian, Xiaoyan Liu, Yuwu Jiang, Yuehua Zhang
ObjectiveTo elucidate the characteristics of the myoclonic seizures alone, or predominant myoclonus combined with generalized tonic-clonic seizures (GTCS) and/or absences, in early childhood, and discuss its classification.MethodsForty-two children were retrospectively recruited between January 2006 and June 2015.ResultsThe mean age of seizure onset was 40.5 months. They were divided into 4 groups: myoclonic seizures alone; predominant myoclonus combined with GTCS; predominant myoclonus combined with absences; predominant myoclonus combined with both GTCS and absences. Interictal EEG showed generalized spike- or polyspike-wave discharges at 2-4 Hz. Seizures were controlled in 22 patients at a mean age of 60.5 months. The psychomotor development was normal (30/37) or mildly delayed (7/37).ConclusionsWe reported a cohort of patients with early childhood myoclonic epilepsy (ECME), with the following characteristics: Seizures started below 5 years old in otherwise normal children; Seizure types included myoclonic seizures alone or combined with GTCS and/or absences; Febrile or afebrile GTCS might appear firstly; Interictal EEG showed generalized spike- or polyspike-wave; Seizures usually were in remission before adolescence with normal development or mild cognitive or behavioral deficits in most.SignificanceECME might be an independent epileptic syndrome not established by International League Against Epilepsy (ILAE) previously.
http://ift.tt/2u07f2B
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Summary Insulinomas are rare neuroendocrine tumours that classically present with fasting hypoglycaemia. This case report discusses an un...
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The online platform for Taylor & Francis Online content New for Canadian Journal of Remote Sen...