bluuebunny/crossref_metadata_2025 huggingface dataset:
Neuromodulation‐dependent effect of gated high‐frequency, LFMS‐like electric field stimulation in mouse cortical slices
Authors: Negahbani Ehsan , Schmidt Stephen L. , Mishal Nadia , Fröhlich Flavio
Published: November 2018

Low‐field magnetic stimulation (LFMS) is a gated high‐frequency non‐invasive brain stimulation method (500 Hz gated at 2 Hz) with a proposed antidepressant effect. However, it has remained unknown how such stimulation paradigms modulate neuronal network activity and how the induced changes depend on network state. Here we examined the immediate and outlasting effects of the gated high‐frequency electric field associated with LFMS on the cortical activity as a function of neuromodulatory tone that defines network state. We used a sham‐controlled study design to investigate effects of stimulation (20 min of 0.5 s trains of 500 Hz charge‐balanced pulse stimulation patterned at 0.5 Hz) on neural activity in mouse medial prefrontal cortex in vitro. Bath application of cholinergic and noradrenergic agents enabled us to examine the stimulation effects as a function of neuromodulatory tone. The stimulation attenuated the increase in firing rate of layer V cortical neurons during the post‐stimulation period in the presence of cholinergic activation. The same stimulation had no significant immediate or outlasting effect in the absence of exogenous neuromodulators or in the presence of noradrenergic activation. These results provide electrophysiological insights into the neuromodulatory‐dependent effects of gated high‐frequency stimulation. More broadly, our results are the first to provide a mechanistic demonstration of how behavioral states and arousal levels may modify the effects of non‐invasive brain stimulation.
Vagus nerve spiking activity associated with locomotion and cortical arousal states in a freely moving rat
Authors: Shikano Yu , Nishimura Yuya , Okonogi Toya , Ikegaya Yuji , Sasaki Takuya
Published: November 2018

The vagus nerve serves as a central pathway for communication between the central and peripheral organs. Despite traditional knowledge of vagus nerve functions, detailed neurophysiological dynamics of the vagus nerve in naïve behavior remain to be understood. In this study, we developed a new method to record spiking patterns from the cervical vagus nerve while simultaneously monitoring central and peripheral organ bioelectrical signals in a freely moving rat. When the rats transiently elevated locomotor activity, the frequency of vagus nerve spikes was correspondingly increased, and this activity was retained for several seconds after the increase in running speed terminated. Spike patterns of the vagus nerve were not robustly associated with which arms the animals entered on an elevated plus maze. During sniffing behavior, vagus nerve spikes were nearly absent. During stopping, the vagus nerve spike patterns differed considerably depending on external contexts and peripheral activity states associated with cortical arousal levels. Stimulation of the vagus nerve altered rat's running speed and cortical arousal states depending on running speed at the instant of stimulation. These observations are a new step for uncovering the physiological dynamics of the vagus nerve modulating the visceral organs such as cardiovascular, respiratory, and gastrointestinal systems.
The importance of growing up: juvenile environment influences dispersal of individuals and their neighbours
Authors: Endriss Stacy B. , Vahsen Megan L. , Bitume Ellyn V. , Grey Monroe J. , Turner Kathryn G. , Norton Andrew P. , Hufbauer Ruth A.
Published: November 2018

Dispersal is a key ecological process that is strongly influenced by both phenotype and environment. Here, we show that juvenile environment influences dispersal not only by shaping individual phenotypes, but also by changing the phenotypes of neighbouring conspecifics, which influence how individuals disperse. We used a model system (Tribolium castaneum, red flour beetles) to test how the past environment of dispersing individuals and their neighbours influences how they disperse in their current environment. We found that individuals dispersed especially far when exposed to a poor environment as adults if their phenotype, or even one‐third of their neighbours’ phenotypes, were shaped by a poor environment as juveniles. Juvenile environment therefore shapes dispersal both directly, by influencing phenotype, as well as indirectly, by influencing the external social environment. Thus, the juvenile environment of even a minority of individuals in a group can influence the dispersal of the entire group.
Effect of glycerol on reconstructed human oral mucosa
Authors: Kvalheim Siri F. , Xenaki Victoria , Kvalheim Arild , Lie Stein‐Atle , Marthinussen Mihaela C. , Strand Gunhild V. , Costea Daniela E.
Published: November 2018

The majority of severely ill patients experience dry mouth. For institutionalized patients, this condition is commonly treated using glycerol as a lubricant. However, because of its possibly desiccating effect, some countries do not advocate the use of glycerol. This study aimed to investigate dose‐dependent effects of glycerol on homeostasis and tissue integrity of in vitro‐reconstructed normal human buccal mucosa (RNHBM). Primary keratinocytes and fibroblasts were isolated and expanded from biopsies of mucosa from eight healthy volunteers. Ninety‐six samples of RNHBM were prepared and exposed for 24 h to 17%, 42.5%, or 85% glycerol, or to distilled H2O (control). Sections were stained with haematoxylin and eosin (H&E) to evaluate epithelial thickness or used for immunohistochemistry to measure expression of Ki67 (proliferation), cleaved caspase‐3 (apoptosis), and E‐cadherin (tissue‐integrity). Positive cells and cell layers, as detected by immunohistochemistry, were counted. Epithelial thickness, proliferation, and apoptosis were significantly increased by exposure to 42.5% and 85% glycerol. No significant differences in apoptosis or proliferation were found between controls and RNHBM exposed to 17% glycerol. E‐cadherin expression was not significantly affected by exposure to any of the concentrations of glycerol tested. This study shows that glycerol affects tissue homeostasis, but not tissue integrity, of RNHBM at glycerol concentrations above 42.5%.
Defining the electroclinical phenotype and outcome of PCDH19‐related epilepsy: A multicenter study
Authors: Trivisano Marina , Pietrafusa Nicola , Terracciano Alessandra , Marini Carla , Mei Davide , Darra Francesca , Accorsi Patrizia , Battaglia Domenica , Caffi Lorella , Canevini Maria P. , Cappelletti Simona , Cesaroni Elisabetta , de Palma Luca , Costa Paola , Cusmai Raffaella , Giordano Lucio , Ferrari Annarita , Freri Elena , Fusco Lucia , Granata Tiziana , Martino Tommaso , Mastrangelo Massimo , Bova Stefania M. , Parmeggiani Lucio , Ragona Francesca , Sicca Federico , Striano Pasquale , Specchio Luigi M. , Tondo Ilaria , Zambrelli Elena , Zamponi Nelia , Zanus Caterina , Boniver Clementina , Vecchi Marilena , Avolio Carlo , Dalla Bernardina Bernardo , Bertini Enrico , Guerrini Renzo , Vigevano Federico , Specchio Nicola
Published: November 2018

Objective

PCDH19‐related epilepsy is an epileptic syndrome with infantile onset, characterized by clustered and fever‐induced seizures, often associated with intellectual disability (ID) and autistic features. The aim of this study was to analyze a large cohort of patients with PCDH19‐related epilepsy and better define the epileptic phenotype, genotype‐phenotype correlations, and related outcome‐predicting factors.

Methods

We retrospectively collected genetic, clinical, and electroencephalogram (EEG) data of 61 patients with PCDH19‐related epilepsy followed at 15 epilepsy centers. All consecutively performed EEGs were analyzed, totaling 551. We considered as outcome measures the development of ID, autistic spectrum disorder (ASD), and seizure persistence. The analyzed variables were the following: gender, age at onset, age at study, genetic variant, fever sensitivity, seizure type, cluster occurrence, status epilepticus, EEG abnormalities, and cognitive and behavioral disorders. Receiver operating characteristic curve analysis was performed to evaluate the age at which seizures might decrease in frequency.

Results

At last follow‐up (median = 12 years, range = 1.9‐42.1 years), 48 patients (78.7%) had annual seizures/clusters, 13 patients (21.3%) had monthly to weekly seizures, and 12 patients (19.7%) were seizure‐free for ≥2 years. Receiver operating characteristic analysis showed a significant decrease of seizure frequency after the age of 10.5 years (sensitivity = 81.0%, specificity = 70.0%). Thirty‐six patients (59.0%) had ID and behavioral disturbances. ASD was present in 31 patients. An earlier age at epilepsy onset emerged as the only predictive factor for ID (P = 0.047) and ASD (P = 0.014). Conversely, age at onset was not a predictive factor for seizure outcome (P = 0.124).

Significance

We found that earlier age at epilepsy onset is related to a significant risk for ID and ASD. Furthermore, long‐term follow‐up showed that after the age of 10 years, seizures decrease in frequency and cognitive and behavioral disturbances remain the primary clinical problems.
Attendance to egg clutches by male Kurixalus eiffingeri increases hatching success and decreases predation by invasive slugs (Parmarion martensi) in Taiwan
Authors: Chuang Ming‐Feng , Borzée Amaël , Kam Yeong‐Choy
Published: November 2018

Invasive species have a significant impact on amphibians, and most notably Chytrid fungi together with a few vertebrates. However, invasive terrestrial invertebrates are seldomly demonstrated to have a negative effect on their host environment, and few studies investigated the behavioural interactions between invasive and local species, limiting our knowledge of the adaptive response adopted by local organisms. These responses include whether parenting animals recognize predators as threats to their offspring and whether parental care is adequately corrected. Here, through manipulative experiments on a population of phytotelm‐breeder Kurixalus eiffingeri in central Taiwan, we demonstrated the positive effects of parental care against predation by the invasive slug Parmarion martensi, and against mould infection. Treatment groups, in the absence of caring males, displayed a higher rate of predation by slugs and mould infections. The presence of the caring males was linked to an increase in hatching success in control groups, suggesting hydration by the males, while skin peptides may be the factor limiting mould infection. We recommend further research on the impact of the invasive but widespread P. martensi on amphibian populations.
Prospective study of blackthorn injury and synovitis in 35 horses
Authors: Ashton N. M.
Published: November 2018

The objective of this prospective clinical study was to investigate the cause and describe the presentation, diagnosis, treatment techniques and outcome of Prunus spinosa (blackthorn) injury and synovitis in the horse. In all cases presented with blackthorn injury and synovitis, surgical treatment was performed within 24 h, using a two‐stage procedure: 1‐Perisynovial technique using ultrasound guided electrosurgical dissection; 2‐Endoscopic technique. The diagnosis was confirmed by retrieval of black plant material from or close to the affected synovial structure. Mean lameness score on presentation was 4/5 (range 1–5). The most commonly affected structures were extensor tendon sheaths (12/35) and fetlock joints (11/35). All cases had thorn material removed, 80% had thorn material removed at surgery and in 49% it was intra‐synovial. On presentation, the mean synovial fluid total protein level (TP) was 47.6 g/L (range 18–66); mean total nucleated cell count (TNCC) was 176 × 109 cells/L (range 12–312). Two days post‐surgery, mean total protein levels were 33 g/L (range 16–52), mean TNCC was 13 × 109 cells/L (range 1–35). At 5 days post‐surgery, the mean total protein was 23 g/L (range 12–28) and TNCC was 5 × 109 cells/L (range 1–12). All synovial fluid cultures were negative. Twenty‐eight (80%) horses were sound 5 days post‐operatively, seven (20%) were not lame in walk; they all returned to full work in an average time of 8 weeks (range 3–48 weeks). Surgery achieved accurate identification and removal of thorn material. In contrast to previous studies of synovial sepsis, these cases had a positive outcome despite high pre‐ and post‐operative synovial fluid TP and TNCC. These findings suggest that Prunus spinosus (blackthorn) synovitis has a different aetiology to synovitis originating from sepsis or other types of contamination.
Increased oxidative stress and CaMKII activity contribute to electro‐mechanical defects in cardiomyocytes from a murine model of Huntington's disease
Authors: Joviano‐Santos Julliane Vasconcelos , Santos‐Miranda Artur , Botelho Ana Flávia Machado , de Jesus Itamar Couto Guedes , Andrade Jéssica Neves , de Oliveira Barreto Tatiane , Magalhães‐Gomes Matheus Proença S. , Valadão Priscila Aparecida Costa , Cruz Jader dos Santos , Melo Marília Martins , Guatimosim Silvia , Guatimosim Cristina
Published: November 2018

Huntington's disease (HD) is a neurodegenerative genetic disorder. Although described as a brain pathology, there is evidence suggesting that defects in other systems can contribute to disease progression. In line with this, cardiovascular defects are a major cause of death in HD. To date, relatively little is known about the peripheral abnormalities associated with the disease. Here, we applied a range of assays to evaluate cardiac electro‐mechanical properties in vivo, using a previously characterized mouse model of HD (BACHD), and in vitro, using cardiomyocytes isolated from the same mice. We observed conduction disturbances including QT interval prolongation in BACHD mice, indicative of cardiac dysfunction. Cardiomyocytes from these mice demonstrated cellular electro‐mechanical abnormalities, including a prolonged action potential, arrhythmic contractions, and relaxation disturbances. Cellular arrhythmia was accompanied by an increase in calcium waves and increased Ca2+/calmodulin‐dependent protein kinase II activity, suggesting that disruption of calcium homeostasis plays a key part. We also described structural abnormalities in the mitochondria of BACHD‐derived cardiomyocytes, indicative of oxidative stress. Consistent with this, imbalances in superoxide dismutase and glutathione peroxidase activities were detected. Our data provide an in vivo demonstration of cardiac abnormalities in HD together with new insights into the cellular mechanistic basis, providing a possible explanation for the higher cardiovascular risk in HD.
Modelled net carbon gain responses to climate change in boreal trees: Impacts of photosynthetic parameter selection and acclimation
Authors: Stinziano Joseph R. , Bauerle William L. , Way Danielle A.
Published: November 2018

Boreal forests are crucial in regulating global vegetation‐atmosphere feedbacks, but the impact of climate change on boreal tree carbon fluxes is still unclear. Given the sensitivity of global vegetation models to photosynthetic and respiration parameters, we determined how predictions of net carbon gain (C‐gain) respond to variation in these parameters using a stand‐level model (MAESTRA). We also modelled how thermal acclimation of photosynthetic and respiratory temperature sensitivity alters predicted net C‐gain responses to climate change. We modelled net C‐gain of seven common boreal tree species under eight climate scenarios across a latitudinal gradient to capture a range of seasonal temperature conditions. Physiological parameter values were taken from the literature together with different approaches for thermally acclimating photosynthesis and respiration. At high latitudes, net C‐gain was stimulated up to 400% by elevated temperatures and CO2 in the autumn but suppressed at the lowest latitudes during midsummer under climate scenarios that included warming. Modelled net C‐gain was more sensitive to photosynthetic capacity parameters (Vcmax, Jmax, Arrhenius temperature response parameters, and the ratio of Jmax to Vcmax) than stomatal conductance or respiration parameters. The effect of photosynthetic thermal acclimation depended on the temperatures where it was applied: acclimation reduced net C‐gain by 10%–15% within the temperature range where the equations were derived but decreased net C‐gain by 175% at temperatures outside this range. Thermal acclimation of respiration had small, but positive, impacts on net C‐gain. We show that model simulations are highly sensitive to variation in photosynthetic parameters and highlight the need to better understand the mechanisms and drivers underlying this variability (e.g., whether variability is environmentally and/or biologically driven) for further model improvement.
Transgenerational lipid‐reducing activity of benzylisoquinoline alkaloids in Caenorhabditis elegans
Authors: Chow Yit‐Lai , Sato Fumihiko
Published: November 2018

Epigenetic mechanisms allow for transgenerational memory of an ancestor's environment and can affect the gene expression, physiology and phenotype of that ancestor's descendants, independent of DNA sequence alteration. Among many model organisms, Caenorhabditis elegans has been instrumental in studies of transgenerational inheritance, most of which have focused on the effects of external stressors of the parent worm on the life span and stress resistance of future generations. In this work, we used Nile red staining of accumulated lipids in C. elegans to investigate the transgenerational effect of two benzylisoquinoline alkaloids, namely, berberine and sanguinarine. Our results showed that a reduction in Nile red fluorescence can be propagated to subsequent worm generations. Using mutant worms, we found that the transgenerational effect requires the ASH‐2 component of the histone H3K4me3 complex and the HRDE‐1 worm Argonaute protein. Ash‐2 is also required for transgenerational inheritance of the xenobiotic response in the worm. Our study offers new insights into transmissible drug effects across multiple generations and suggests the importance of such analyses in the drug development process.
Caries severity declined besides persistent untreated primary teeth over a 22‐year period: Trends among children in Goiânia, Brazil
Authors: Freire Maria do Carmo Matias , Daher Anelise , Costa Luciane Rezende , Corrêa‐Faria Patrícia , de Brito Letícia Candine , Bönecker Marcelo José Strazzeri , de Abreu Mauro Henrique Nogueira Guimarães
Published: November 2018

Background

Evidence of time trends in early childhood caries in low‐ and middle‐income countries in the second decade of the year 2000 is scarce.

Aim

To assess the trends in early childhood caries prevalence and severity in 2‐ to 5‐year‐old children over a 22‐year period (1993‐2015) in Goiânia, Midwest Brazil. Additionally, we aimed to investigate the changes regarding affected dental arches and teeth.

Design

A time‐lag analysis of trends in caries was carried out using data from three cross‐sectional studies based on the World Health Organization diagnostic criteria, in 1993 (N = 1362), 2001 (N = 1620), and 2015 (N = 548).

Results

Caries prevalence declined from 45.1% in 1993 to 29.0% in 2015. Prevalence of severe caries (dmft ≥ 6) in 2015 was nearly one‐third of that found in 1993, and the SiC index (mean dmft of the highest tertile) decreased from 4.55 to 3.32. Decline was higher in the 1993‐2001 than in the 2001‐2015 time‐lag. Posterior teeth and second molars had the greatest reductions. High proportions of untreated caries were found in all ages and survey years.

Conclusion

There were significant changes in caries prevalence and severity, marked by a striking decline from 1993 to 2001, followed by a less prominent decrease up to 2015, and high levels of untreated caries.
Efficacy of a socioemotional learning programme in a sample of children with intellectual disability
Authors: Faria Sónia M. M. , Esgalhado Graça , Pereira Cristina M. G.
Published: November 2018

Socioemotional learning (SEL) is the process of developing social and emotional competences through which the child learns to recognize and manage emotions, care for others, make good decisions, behave responsibly and ethically, develop positive relationships and avoid negative behaviours (Promoting social and emotional learning: Guidelines for educators, 1997). To promote the development of these competences, specific programmes are developed. This study aims to analyse the impact of a SEL programme among children with intellectual disability. Two groups were established, control and experimental, and the Test of Emotion Comprehension (TEC) (Test of emotion comprehension – TEC, 2000; Les émotions à l'école, émotions et apprentissage, 2004) was applied to both as pre‐ and post‐test. The programme was implemented in the experimental group, based on simply illustrated stories alluding the basic emotions. The findings suggest that the programme is effective in the development of socioemotional competences (SEC) in subjects with intellectual disabilities, evidencing the positive effects that the intervention programme had with the experimental group.
TSG101 promotes the proliferation, migration and invasion of hepatocellular carcinoma cells by regulating the PEG10
Authors: Liu Zhiyi , Tian Zilu , Cao Kuan , Zhang Bin , Wen Quan , Zhou Xinyu , Yang Weibin , Wang Tao , Shi Hengliang , Wang Renhao
Published: November 2018

The tumour susceptibility gene 101 (TSG101) is reported to play important roles in the development and progression of several human cancers. However, its potential roles and underlined mechanisms in human hepatocellular carcinoma (HCC) are still needed to be further clarified. In the present study, we reported that knock down of TSG101 suppressed the proliferation, migration and invasion of HCC cells, while overexpression of TSG101 facilitated them. Molecularly, the results revealed that knock down of TSG101 significantly decreased the cell cycle related regulatory factor p53 and p21. In another point, knock down of TSG101 also obviously decreased the level of metallopeptidase inhibitor TIMP1 (Tissue inhibitors of metalloproteinases 1), which results in inhibition of MMP2, MMP7 and MMP9. In contrast, overexpression of TSG101 had opposite effects. The iTRAQ proteomics analysis identified that oncogenic protein PEG10 (Paternally expressed gene 10) might be a potential downstream target of TSG101. Further investigation showed that TSG101 interacted with PEG10 and protected it from proteasomal degradation thereby regulating the expression of p53, p21 and MMPs. Finally, we found that both TSG101 and PEG10 proteins are up‐regulated and presented a direct correlation in HCC patients. In conclusion, these results suggest that TSG101 is up‐regulated in human HCC patients, which may accelerate the proliferation, migration and invasion of HCC cells through regulating PEG10.
Pharmacological effects of berberine on mood disorders
Authors: Fan Jie , Zhang Kun , Jin Yang , Li Bingjini , Gao Shuohui , Zhu Jiaming , Cui Ranji
Published: November 2018

Berberine, a natural isoquinoline alkaloid, is used in herbal medicine and has recently been shown to have efficacy in the treatment of mood disorders. Furthermore, berberine modulates neurotransmitters and their receptor systems within the central nervous system. However, the detailed mechanisms of its action remain unclear. This review summarizes the pharmacological effects of berberine on mood disorders. Therefore, it may be helpful for potential application in the treatment of mood disorders.
Efficient method for isolation of reticulocyte RNA from healthy individuals and hemolytic anaemia patients
Authors: Skulski Michał , Bartoszewski Rafał , Majkowski Michał , Machnicka Beata , Kuliczkowski Kazimierz , Sikorski Aleksander F. , Bogusławska Dżamila M.
Published: November 2018

Despite enormous progress and development of high‐throughput methods in genome‐wide mRNA analyses, data on the erythroid transcriptome are still limited, even though they could be useful in medical diagnostics and personalized therapy as well as in research on normal and pathological erythroid maturation. Although obtaining normal and pathological reticulocyte transcriptome profiles should contribute greatly to our understanding of the molecular bases of terminal erythroid differentiation as well as the mechanisms of the hematological diseases, a basic limitation of these studies is the difficulty of efficient reticulocyte RNA isolation from human peripheral blood. The restricted number of possible parallel experiments primarily concern healthy individuals with the lowest number of reticulocytes in the peripheral blood and a low RNA content. In the present study, an efficient method for reticulocyte RNA isolation from healthy individuals and hemolytic anaemia patients is presented. The procedure includes leukofiltration, Ficoll‐Paque gradient centrifugation, Percoll gradient centrifugation, and negative (CD45 and CD61) immunomagnetic separation. This relatively fast and simple four‐stage method was successfully applied to obtain a reticulocyte‐rich population from healthy subjects, which was used to efficiently isolate the high‐quality RNA essential for successful NGS‐based transcriptome analysis.
DLG5 suppresses breast cancer stem cell‐like characteristics to restore tamoxifen sensitivity by inhibiting TAZ expression
Authors: Liu Jie , Li Juan , Li Pingping , Jiang Yina , Chen He , Wang Ruiqi , Cao Fang , Liu Peijun
Published: November 2018

Tamoxifen (TAM) is a primary drug for treatment of estrogen receptor positive breast cancer. However, TAM resistance remains a serious threat to breast cancer patients and may be attributed to increased stemness of breast cancer. Here, we show that discs large homolog 5 (DLG5) expression is down‐regulated in TAM‐resistant breast cancer and cells. DLG5 silencing decreased the sensitivity to TAM and increased the frequency and stemness of CD44+/CD24− breast cancer stem cells (BCSCs) and TAZ, a transducer of the Hippo pathway, expression in MCF7 cells while DLG5 overexpression had opposite effects. TAZ silencing restored the sensitivity to TAM and reduced the frequency and stemness in TAM‐resistant breast cancer cells. Taken together, our data indicate that down‐regulated DLG5 expression increases the stemness of breast cancer cells by enhancing TAZ expression, contributing to TAM resistance in breast cancer.
Evodiamine inhibits RANKL‐induced osteoclastogenesis and prevents ovariectomy‐induced bone loss in mice
Authors: Jin Haiming , Yao Lingya , Chen Kai , Liu Yuhao , Wang Qingqing , Wang Ziyi , Liu Qian , Cao Zhen , Kenny Jacob , Tickner Jennifer , Wang Xiangyang , Xu Jiake
Published: November 2018

Postmenopausal osteoporosis (PMO) is a progressive bone disease characterized by the over‐production and activation of osteoclasts in elderly women. In our study, we investigated the anti‐osteoclastogenic effect of evodiamine (EVO) in vivo and in vitro, as well as the underlying mechanism. By using an in vitro bone marrow macrophage (BMM)‐derived osteoclast culture system, we found that EVO inhibited osteoclast formation, hydroxyapatite resorption and receptor activator of NF‐κB ligand (RANKL)‐induced osteoclast marker gene and protein expression. Mechanistically, we found that EVO inhibited the degradation and RANKL‐induced transcriptional activity of IκBα. RANKL‐induced Ca2+ oscillations were also abrogated by EVO. In vivo, an ovariectomized (OVX) mouse model was established to mimic PMO, and OVX mice received oral administration of either EVO (10 mg/kg) or saline every other day. We found that EVO can attenuate bone loss in OVX mice by inhibiting osteoclastogenesis. Taken together, our findings suggest that EVO suppresses RANKL‐induced osteoclastogenesis through NF‐κB and calcium signalling pathways and has potential value as a therapeutic agent for PMO.
Immunometabolism features of metabolic deregulation and cancer
Authors: Wang Xue , Ping Feng‐Feng , Bakht Sahar , Ling Jingjing , Hassan Waseem
Published: November 2018

Immunometabolism is a branch dealing at the interface of immune functionalities and metabolic regulations. Considered as a bidirectional trafficking, metabolic contents and their precursors bring a considerable change in immune cells signal transductions which as a result affect the metabolic organs and states as an implication. Lipid metabolic ingredients form a major chunk of daily diet and have a proven contribution in immune cells induction, which then undergo metabolic pathway shuffling inside their ownself. Lipid metabolic states activate relevant metabolic pathways inside immune cells that in turn prime appropriate responses to outside environment in various states including lipid metabolic disorders itself and cancers as an extension. Although data on Immunometabolism are still growing, but scientific community need to adjust and readjust according to recent data on given subject. This review attempts to provide current important data on Immunometabolism and consequently its metabolic ramifications. Incumbent data on various lipid metabolic deregulations like obesity, metabolic syndrome, obese asthma and atherosclerosis are analysed. Further, metabolic repercussions on cancers and its immune modalities are also analysed.
Age‐related ultrastructural changes of the basement membrane in the mouse blood‐brain barrier
Authors: Ceafalan Laura Cristina , Fertig Tudor Emanuel , Gheorghe Teodora Cristina , Hinescu Mihail Eugen , Popescu Bogdan Ovidiu , Pahnke Jens , Gherghiceanu Mihaela
Published: November 2018

The blood‐brain barrier (BBB) is essential for a functional neurovascular unit. Most studies focused on the cells forming the BBB, but very few studied the basement membrane (BM) of brain capillaries in ageing. We used transmission electron microscopy and electron tomography to investigate the BM of the BBB in ageing C57BL/6J mice. The thickness of the BM of the BBB from 24‐month‐old mice was double as compared with that of 6‐month‐old mice (107 nm vs 56 nm). The aged BBB showed lipid droplets gathering within the BM which further increased its thickness (up to 572 nm) and altered its structure. The lipids appeared to accumulate toward the glial side of the BM. Electron tomography showed that the lipid‐rich BM regions are located in small pockets formed by the end‐feet of astrocytes. These findings suggest an imbalance of the lipid metabolism and that may precede the structural alteration of the BM. These alterations may favour the accretion of abnormal proteins that lead to neurodegeneration in ageing. These findings warrant further investigation of the BM of brain capillaries and of adjoining cells as potential targets for future therapies.
High proportion of transient neonatal zinc deficiency causing alleles in the general population
Authors: Golan Yarden , Lehvy Adrian , Horev Guy , Assaraf Yehuda G.
Published: November 2018

Loss of function (LoF) mutations in the zinc transporter SLC30A2/ZnT2 result in impaired zinc secretion into breast milk consequently causing transient neonatal zinc deficiency (TNZD) in exclusively breastfed infants. However, the frequency of TNZD causing alleles in the general population is yet unknown. Herein, we investigated 115 missense SLC30A2/ZnT2 mutations from the ExAC database, equally distributed in the entire coding region, harboured in 668 alleles in 60 706 healthy individuals of diverse ethnicity. To estimate the frequency of LoF SLC30A2/ZnT2 mutations in the general population, we used bioinformatics tools to predict the potential impact of these mutations on ZnT2 functionality, and corroborated these predictions by a zinc transport assay in human MCF‐7 cells. We found 14 missense mutations that were markedly deleterious to zinc transport. Together with two conspicuous LoF mutations in the ExAC database, 26 SLC30A2/ZnT2 alleles harboured deleterious mutations, suggesting that at least 1 in 2334 newborn infants are at risk to develop TNZD. This high frequency of TNZD mutations combined with the World Health Organization‐promoted increase in the rate of exclusive breastfeeding highlights the importance of genetic screening for inactivating SLC30A2/ZnT2 mutations in the general population for the early diagnosis and prevention of TNZD.
Otx2 enhances transdifferentiation of Müller cells‐derived retinal stem cells into photoreceptor‐like cells
Authors: Xiong Yu , Ji Hongpei , You Zhipeng , Yao Fei , Zhou Rongrong , Song Weitao , Xia Xiaobo
Published: November 2018

Retinal Müller glial cells have the potential of neurogenic retinal progenitor cells, and could reprogram into retinal‐specific cell types such as photoreceptor cells. How to promote the differentiation of Müller cells into photoreceptor cells represents a promising therapy strategy for retinal degeneration diseases. This study aimed to enhance the transdifferentiation of rat Müller cells‐derived retinal stem cells (MC‐RSCs) into photoreceptor‐like cells and explore the signalling mechanism. We dedifferentiated rat Müller cells into MC‐RSCs which were infected with Otx2 overexpression lentivirus or control. The positive rate of photoreceptor‐like cells among MC‐RSCs treated with Otx2 overexpression lentivirus was significantly higher compared to control. Furthermore, pre‐treatment with Crx siRNA, Nrl siRNA, or GSK‐3 inhibitor SB‐216763 reduced the positive rate of photoreceptor‐like cells among MC‐RSCs treated with Otx2 overexpression lentivirus. Finally, Otx2 induced photoreceptor precursor cells were injected into subretinal space of N‐methyl‐N‐nitrosourea induced rat model of retinal degeneration and partially recovered retinal degeneration in the rats. In conclusion, Otx2 enhances transdifferentiation of MC‐RSCs into photoreceptor‐like cells and this is associated with the inhibition of Wnt signalling. Otx2 is a potential target for gene therapy of retinal degenerative diseases.
Mast cell tetrahydrobiopterin contributes to itch in mice
Authors: Zschiebsch Katja , Fischer Caroline , Wilken‐Schmitz Annett , Geisslinger Gerd , Channon Keith , Watschinger Katrin , Tegeder Irmgard
Published: November 2018

GTP cyclohydrolase (GCH1) governs de novo synthesis of the enzyme cofactor, tetrahydrobiopterin (BH4), which is essential for biogenic amine production, bioactive lipid metabolism and redox coupling of nitric oxide synthases. Overproduction of BH4 via upregulation of GCH1 in sensory neurons is associated with nociceptive hypersensitivity in rodents, and neuron‐specific GCH1 deletion normalizes nociception. The translational relevance is revealed by protective polymorphisms of GCH1 in humans, which are associated with a reduced chronic pain. Because myeloid cells constitute a major non‐neuronal source of BH4 that may contribute to BH4‐dependent phenotypes, we studied here the contribution of myeloid‐derived BH4 to pain and itch in lysozyme M Cre‐mediated GCH1 knockout (LysM‐GCH1−/−) and overexpressing mice (LysM‐GCH1‐HA). Unexpectedly, knockout or overexpression in myeloid cells had no effect on nociceptive behaviour, but LysM‐driven GCH1 knockout reduced, and its overexpression increased the scratching response in Compound 48/80 and hydroxychloroquine‐evoked itch models, which involve histamine and non‐histamine dependent signalling pathways. Mechanistically, GCH1 overexpression increased BH4, nitric oxide and hydrogen peroxide, and these changes were associated with increased release of histamine and serotonin and degranulation of mast cells. LysM‐driven GCH1 knockout had opposite effects, and pharmacologic inhibition of GCH1 provided even stronger itch suppression. Inversely, intradermal BH4 provoked scratching behaviour in vivo and BH4 evoked an influx of calcium in sensory neurons. Together, these loss‐ and gain‐of‐function experiments suggest that itch in mice is contributed by BH4 release plus BH4‐driven mediator release from myeloid immune cells, which leads to activation of itch‐responsive sensory neurons.
Unveiling Ga(III) phthalocyanine—a different photosensitizer in neuroblastoma cellular model
Authors: Constantin Carolina , Lupu Andreea‐Roxana , Fertig Tudor Emanuel , Gherghiceanu Mihaela , Pop Sevinci , Ion Rodica‐Mariana , Neagu Monica
Published: November 2018

Phthalocyanines (Pc) and their metallated derivatives are strongly considered for photodynamic therapy (PDT) possessing unique properties as possible new photosensitizers (PS). We have used toxicological assessments, real‐time monitoring of cellular impedance, and imagistic measurements for assessing the in vitro dark toxicity and PDT efficacy of Ga(III)‐Pc in SHSy5Y neuroblastoma cells. We have established the non‐toxic concentration range of Ga(III)‐Pc, a compound which shows a high intracellular accumulation, with perinuclear distribution in confocal microscopy. By choosing Ga(III)Pc non‐toxic dose, we performed in vitro experimental PDT hampering cellular proliferation. Our proposed Ga(III)‐Pc could complete a future PS panel for neuroblastoma alternate therapy.
Intracellular reduction in ATP levels contributes to CYT997‐induced suppression of metastasis of head and neck squamous carcinoma
Authors: Zhao Xiangdong , Lang Liwei , He Leilei , Gao Lixia , Chyan David , Xiong Yuanping , Li Honglin , Peng Hong , Teng Yong
Published: November 2018

The incidence rate of head and neck squamous cell carcinoma (HNSCC) has steadily increased over the past decade. However, treatment options for metastatic HNSCC are often limited and the 5‐year survival rate has remained static. Therefore, the development and assessment of more efficient but less toxic therapeutic strategies is an unmet need for treatment of more extensive HNSCC. Here, we report that CYT997, a novel microtubule‐disrupting agent, exerts strong activity in inhibiting HNSCC cell invasion and metastasis. The loss of invasion capacity by CYT997 was accompanied by an associated increase in cell adhesion and the reversal of epithelial‐mesenchymal transition (EMT). Increased expression of E‐cadherin protein and decreased expression of Vimentin protein became evident in HNSCC cells following CYT997 exposure, which were consistently observed in HNSCC xenografts from the mice receiving CYT997. Moreover, the capacity of invasive HNSCC cells to form pulmonary metastases was significantly blocked with CYT997 treatment, indicating that the diminishment of EMT traits contributes to CYT997‐suppressed metastasis. Intriguingly, CYT997 impaired intracellular ATP levels in HNSCC cells, at least in part, through its inhibitory effect on the mitochondrial protein IF1. The addition of ATP attenuated CYT997‐induced suppression of cell invasion, coupled with down‐regulation of E‐Cadherin and up‐regulation of Vimentin. These findings support a critical role of ATP levels in cell invasion and metastasis under the influence of CYT997. Collectively, our data unveil the mechanism involved in mediating CYT997 action, and provide preclinical rationale for possible clinical application of CYT997 as a novel therapeutic strategy against aggressive HNSCC.
Two human MARs effectively increase transgene expression in transfected CHO cells
Authors: Li Qin , Zhao Chun‐peng , Lin Yan , Song Chao , Wang Fang , Wang Tian‐yun
Published: November 2018

Matrix attachment regions (MARs) can enhance the expression level of transgene in Chinese hamster ovaries (CHO) cell expression system. However, improvements in function and analyses of the mechanism remains unclear. In this study, we screened two new and more functional MAR elements from the human genome DNA. The human MAR‐3 and MAR‐7 element were cloned and inserted downstream of the polyA site in a eukaryotic vector. The constructs were transfected into CHO cells, and screened under G418 to produce the stably transfected cell pools. The expression levels and stability of enhanced green fluorescent protein (eGFP) were detected by flow cytometry. The transgene copy number and transgene expression at mRNA level were detected by quantitative real‐time PCR. The results showed that the expression level of eGFP of cells transfected with MAR‐containing vectors were all higher than those of the vectors without MARs under transient and stably transfection. The enhancing effect of MAR‐7 was higher than that of MAR‐3. Additionally, we found that MAR significantly increased eGFP copy numbers and eGFP gene mRNA expression level as compared with the vector without. In conclusion, MAR‐3 and MAR‐7 gene can promote the expression of transgene in transfected CHO cells, and its effect may be related to the increase of the number of copies.
Metformin relieves neuropathic pain after spinal nerve ligation via autophagy flux stimulation
Authors: Weng Weidong , Yao Chenglun , Poonit Keshav , Zhou Xijie , Sun Chao , Zhang Feng , Yan Hede
Published: November 2018

Neuropathic pain is a well‐known type of chronic pain caused by damage to the nervous system. Autophagy is involved in the development and/or progression of many diseases, including neuropathic pain. Emerging evidence suggests that metformin relieves neuropathic pain in several neuropathic pain models; however, metformin's cellular and molecular mechanism for pain relief remains unknown. In this study, we investigated the therapeutic effects of metformin on pain relief after spinal nerve ligation (SNL) and its underlying mechanism of autophagy regulation. Behavioural analysis, histological assessment, expression of c‐Fos and molecular biological changes, as well as ultrastructural features, were investigated. Our findings showed that the number of autophagosomes and expression of autophagy markers, such as LC3 and beclin1, were increased, while the autophagy substrate protein p62, as well as the ubiquitinated proteins, were accumulated in the ipsilateral spinal cord. However, metformin enhanced the expression of autophagy markers, while it abrogated the abundance of p62 and ubiquitinated proteins. Blockage of autophagy flux by chloroquine partially abolished the apoptosis inhibition and analgesic effects of metformin on SNL. Taken together, these results illustrated that metformin relieved neuropathic pain through autophagy flux stimulation and provided a new direction for metformin drug development to treat neuropathic pain.
Glucose‐dependent insulinotropic polypeptide deficiency reduced fat accumulation and insulin resistance, but deteriorated bone loss in ovariectomized mice
Authors: Shimazu‐Kuwahara Satoko , Kanemaru Yoshinori , Harada Norio , Ikeguchi Eri , Ueda Yohei , Yamane Shunsuke , Murata Yuki , Yasoda Akihiro , Kieffer Timothy J , Inagaki Nobuya
Published: November 2018

Given the established roles of glucose‐dependent insulinotropic polypeptide (GIP) in promoting fat storage and bone formation, we assessed the contribution of GIP to obesity and osteopenia in ovariectomized mice with a gene encoding green fluorescent protein (GFP) inserted into the GIP locus, in which GIP was either reduced (GIPgfp/+) or absent (GIPgfp/gfp). In GIPgfp/gfp mice, weight gain, subcutaneous and visceral fat mass were reduced, and glucose intolerance was improved compared with wild‐type mice with the same magnitude of insulin responses. Cancellous bone mineral density and bone cortical thickness were reduced in GIPgfp/gfp mice compared with wild‐type mice. In GIPgfp/+ mice, weight gain, glucose intolerance and cancellous bone mineral density were not different from that of wild‐type mice. These results indicate that the total elimination of GIP ameliorates weight gain and adiposity in ovariectomized mice, but it enhances osteopenia, particularly in cancellous bone by partly suppressing bone formation.
Spatial distribution and sampling plan of the phytophagous stink bug complex in different soybean production systems
Authors: Fernandes Marcos Gino , Costa Eduardo Neves , Cavada Leonardo Hiroito , Mota Thiago Alexandre , da Fonseca Paulo Rogério Beltramin
Published: November 2018

Phytophagous stink bugs are major soybean pests, and knowledge of spatial distribution models of the pest in the crop is fundamental to establishing an appropriate sequential sampling plan, and thus, allowing the correct utilization of control strategies. This work aimed to study the spatial distribution of phytophagous stink bugs in soybean grown in different cropping systems and determine a sequential sampling plan. The experiment was conducted in Maracaju, MS, Brazil, during the agricultural year of 2012/2013. Soybean cultivars BRS 284 and SYN 1163 RR were placed in an experimental area comprising six fields (two soybean cultivars × three cropping systems). Sampling was performed weekly, using a beat cloth per plot and counting the number of stink bugs found, virtually throughout the soybean reproductive period. Concerning the statistical analyses, we used the dispersion indexes (variance‐to‐mean ratio, Morisita's index, exponent k of the negative binomial and Green's coefficient) and probabilistic methods of frequency adjustment (negative binomial and Poisson). Adult and nymph phytophagous stink bugs showed aggregate disposition in the field regardless of the cropping system, and their numbers were adjusted to the negative binomial probability distribution. There was no difference in the behaviour of adult and nymphs considering the tested cultivars. We elaborated a practical sequential sampling plan for phytophagous stink bug complexes, considering crops intended for the production of grains and seeds.
Quality of hypertension care: An improvement initiative in two outpatient health care centers
Authors: Yusupov Eleanor , Krishnamachari Bhuma , Rand Sarah , Abdalla Mirette , Zwibel Hallie
Published: November 2018

Rationale, aims, and objectives

Hypertension control is an important public health goal; however, significant barriers remain in primary care practice.

Our objective was to identify areas for improvement in hypertension care and implement changes in management to improve outcomes. We also aimed to evaluate whether quality improvement influences physician attitudes towards and adherence to current hypertension guidelines.

Method

We conducted a non‐experimental pre‐ vs post‐ design quality improvement study for ambulatory patients with a history of hypertension. Specific measures of hypertension care were assessed at baseline and 3 months post‐implementation of the quality improvement initiative.

De‐identified data were collected from 100 charts, randomly selected from the practice's electronic medical records, and compared with a national sample of peer data.

The Intervention was based on the American Academy of Family Physicians METRIC Performance Improvement module. This consisted of creating a computerized registry, system improvements to the electronic medical records, and peer education workshops on best practices.

A 7‐item survey was completed by primary care physicians pre‐ and post‐intervention.

Results

Improvement was demonstrated in several primary outcome measures: increased number of patients counselled on sodium intake (P = 0.005), physical activity (P = 0.001), alcohol consumption (P = 0.03), and weight reduction (P < 0.0001).

Practice self‐assessment findings did not show a statistically significant change following the intervention.

Conclusions

This quality improvement increased provider compliance with hypertension guidelines. However, more effort is required to modify physician practices for full compliance with the 2017 updated hypertension guidelines.