bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of genomic resources for the tick Ixodes ricinus: isolation and characterization of single nucleotide polymorphisms
Authors: Quillery E. , Quenez O. , Peterlongo P. , Plantard O.
Published: October 2013

Assessing the genetic variability of the tick Ixodes ricinus—an important vector of pathogens in Europe—is an essential step for setting up antitick control methods. Here, we report the first identification of a set of SNPs isolated from the genome of I. ricinus, by applying a reduction in genomic complexity, pyrosequencing and new bioinformatics tools. Almost 1.4 million of reads (average length: 528 nt) were generated with a full Roche 454 GS FLX run on two reduced representation libraries of I. ricinus. A newly developed bioinformatics tool (DiscoSnp), which isolates SNPs without requiring any reference genome, was used to obtain 321 088 putative SNPs. Stringent selection criteria were applied in a bioinformatics pipeline to select 1768 SNPs for the development of specific primers. Among 384 randomly SNPs tested by Fluidigm genotyping technology on 464 individuals ticks, 368 SNPs loci (96%) exhibited the presence of the two expected alleles. Hardy–Weinberg equilibrium tests conducted on six natural populations of ticks have shown that from 26 to 46 of the 384 loci exhibited significant heterozygote deficiency.
Disease Modifying Therapies Modulate Cardiovascular Risk Factors in Patients with Multiple Sclerosis
Authors: Sternberg Zohara , Leung Christopher , Sternberg Daniel , Yu Jinhee , Hojnacki David
Published: October 2013

Objectives

This retrospective study aimed to determine (1) the association between the use of three major disease modifying therapies (DMTs) (Interferon‐beta [IFN‐β], Glatiramer acetate [GA], Natalizumab [NTZ]) and cardiovascular (CV) risk factors in multiple sclerosis (MS) patients, and (2) the association between the use of CV drugs (antihypertensive, hypolipidemic, and antiplatelets) and other drugs acting on the CV system (antispastics/anticonvulsants/anxyolitics, antidepressants/stimulants), and MS disease severity.

Methods

The charts of 188 patients with MS, who were taking one of the three DMTs, and 110 patients, who were naïve to these drugs, were retrospectively reviewed. The obtained data included height and weight, fasting lipid profiles, plasma glucose, systolic and diastolic BP, smoking habit, list of medications, and indicators of MS disease severity.

Results

The use of DMTs was associated with higher diastolic BP readings, as well as higher plasma glucose and HDL‐C plasma levels. In addition, there was an association between CV risk factors and the type of DMTs. When compared to DMT‐naïve patients with MS, the use of IFN‐β and GA was associated with higher CV risk factors, whereas the use of NTZ was associated with lower CV risk factors. In DMT‐naïve patients, the use of CV and related drugs was associated with higher Extended Disability Status Scale (EDSS) and higher MS Severity Scale (MSSS).

Conclusion

There is an association between higher CV risk factors and the use of DMTs. Furthermore, CV and related drugs have the potential for modulating MS disease severity.
The status of serum vitamin D in patients with active Behcet's disease compared with controls
Authors: Khabbazi Alireza , Rashtchizadeh Nadereh , Ghorbanihaghjo Amir , Hajialiloo Mehrzad , Ghojazadeh Mortasa , Taei Ramin , Kolahi Sousan
Published: October 2013

Objective

The aim of this study was to characterize the status of vitamin D in patients with active and recently diagnosed Behcet's disease (BD) and the relationship between vitamin D levels and BD activity.

Methods and materials

In this cross sectional study 48 patients with BD and 47 age‐ and sex‐matched healthy controls were included. BD was diagnosed by the International Criteria for BD. Behcet's patients were new cases who were not on any treatment. BD activity was measured by the Iranian Behcet's Disease Dynamic Activity Measure (IBDDAM) and Behcet's Disease Current Activity Form (BDCAF). 25(OH)D measured by enzyme‐linked immunosorbent assay method as an indicator of vitamin D status.

Results

The mean 25‐hydroxyvitamin D (25(OH)D level in the BD group was lower than the control group. Insufficiency and deficiency of 25(OH)D in the BD group was more common than the control group. No correlation was observed between the total IBDDAM, ophthalmic IBDDAM, and BDCAF with 25(OH)D levels. No correlation was found between the major symptoms of BD and 25(OH)D value.

Conclusions

Our study suggests that deficiency of 25(OH)D may be a trigger factor for BD.
Metagenomic analysis on seasonal microbial variations of activated sludge from a full‐scale wastewater treatment plant over 4 years
Authors: Ju Feng , Guo Feng , Ye Lin , Xia Yu , Zhang Tong
Published: October 2013

Metagenomic technique was employed to characterize the seasonal dynamics of activated sludge (AS) communities in a municipal wastewater treatment plant (WWTP) over 4 years. The results indicated that contrary to Eukaryota (mainly Rotifera and Nematoda), abundances of Bacteria and Archaea (mainly Euryarchaeota) were significantly higher in winter than summer. Two‐way analysis of variance and canonical correspondence analysis revealed that many functionally important genera followed strong seasonal variation patterns driven by temperature and salinity gradients; among them, two nitrifying bacteria, Nitrospira and Nitrosomonas, displayed much higher abundances in summer, whereas phosphate‐removing genus Tetrasphaera, denitrifier Paracoccus and potential human faecal bacteria, i.e. Bifidobacterium, Dorea and Ruminococcus, showed significantly higher abundances in winter. Particularly, occurrence of dual variation patterns beyond explanation merely by seasonality indicated that multivariables (e.g. dissolved oxygen, sludge retention time, nutrients) participated in shaping AS community structure. However, SEED subsystems annotation showed that functional categories in AS showed no significant difference between summer and winter, indicating that compared with its microbial components, the functional profiles of AS were much more stable. Taken together, our study provides novel insights into the microbial community variations in AS and discloses their correlations with influential factors in WWTPs.
Eosinophil major basic protein activates human cord blood mast cells primed with fibroblast membranes by integrin‐β1
Authors: Ben‐Zimra M. , Bachelet I. , Seaf M. , Gleich G. J. , Levi‐Schaffer F.
Published: October 2013

Background

Mast cell (MC) – eosinophil (Eos) activating cross‐talk might be critical for the severity and chronicity of allergy. Among soluble mediators, eosinophil major basic protein (MBP), a hallmark of allergy, is particularly important because it was shown to activate specific MC subtypes. We previously demonstrated that MBP activates IgE‐desensitized rat MC and human lung and cord blood‐derived MC (CBMC) after priming with fibroblast membranal stem cell factor. However, a distinct mechanism for this activation was missing. Therefore, we aimed to investigate it.

Methods

Major basic protein‐1 activation of CBMC primed with fibroblast‐derived membranes (FBM) was measured by β‐hexosaminidase and tryptase release. Chemical cross‐linking followed by micrometric flow cytometry probed direct interactions. Antibodies neutralized integrin‐β1 and recognized its active form. Pertussis toxin (Ptx) was used to decrease integrin‐β1 active form expression. Hematopoietic cell kinase (Hck) was identified by immunoprecipitation (IP) and silenced by siRNA.

Results

Major basic protein‐1‐induced CBMC activation is mediated partly by MBP1–integrin‐β1 interaction on the MC surface. FBM prime CBMC via a G protein, as confirmed by Ptx, to shift integrin‐β1 to its active form. Following MBP1 binding, integrin‐β1 binds Hck that further transduces the activation signal. MC priming with FBM leads to up‐regulation in Hck protein level. MC integrin‐β1 neutralization inhibits MBP1‐induced activation and uptake. Hck silencing results with reduced MBP1‐induced activation.

Conclusions

Fibroblast‐derived membranes, integrin‐β1, and Hck are involved in MBP1‐induced activation of CBMC and therefore represent a distinct mechanism for this activation. This finding might implicate integrin‐β1 and Hck as targets for decreasing MC – Eos activating cross‐talk in allergy.
Early Repolarization Syndrome: Electrocardiographic Signs and Clinical Implications
Authors: Rezus Ciprian , Floria Mariana , Moga Victor Dan , Sirbu Oana , Dima Nicoleta , Ionescu Simona Daniela , Ambarus Valentin
Published: October 2013

Early repolarization syndrome (ERS) was previously considered as a benign variant, but it has recently emerged as a risk marker for idiopathic ventricular fibrillation (VF) and sudden death. As measured by electrocardiogram (ECG), early repolarization is characterized by an elevation of the J point and/or ST segment from the baseline by at least 0.1 mV in at least two adjoining leads. In particular, early repolarization detected by inferior ECG leads was found to be associated with idiopathic VF and has been termed as ERS. This condition is mainly observed in young men, athletes, and blacks. Also, it has become evident that electrocardiographic territory, degree of J‐point elevation, and ST‐segment morphology are associated with different levels of risk for subsequent ventricular arrhythmia. However, it is unclear whether J waves are more strongly associated with a depolarization abnormality rather than a repolarization abnormality. Several clinical entities can cause ST‐segment elevation. Therefore, clinical and ECG data are essential for differential diagnosis. At present, the data set is insufficient to allow risk stratification in asymptomatic individuals. ERS, idiopathic VF, and Brugada syndrome (known as J‐wave syndromes) are three clinical conditions that share many common ECG features; however, their clinical consequences are remarkably different. This review summarizes the current electrocardiographic data concerning ERS with clinical implications.
Experience of fetal scalp blood sampling during labor
Authors: Liljeström Lena , Wikström Anna‐Karin , Skalkidou Alkistis , Åkerud Helena , Jonsson Maria
Published: October 2013

Fetal scalp blood sampling (FBS) is often claimed to be painful for women in labor and difficult for obstetricians to perform. Our aim was to assess women's experience of pain during FBS and obstetricians' experience of difficulty in performing the test. At a tertiary center in Sweden, a questionnaire with answers on a 10‐point scale was completed by 51 women and the obstetricians performing the test. Women's experience of pain had a median of 3.5. FBS was well tolerated in women who had epidural analgesia but might be associated with pain in women without. Higher maternal body mass index and less cervical dilation were associated with higher pain ratings. Obstetricians did not generally experience scalp sampling as difficult to perform (median score 3.0). However, the sampling procedure can be more complicated in situations with higher maternal body mass index, less cervical dilation, and a higher station of the fetal head.
Blood eosinophil counts during bronchiolitis are related to bronchial hyper‐responsiveness and lung function in early adolescence
Authors: Mikalsen Ingvild Bruun , Halvorsen Thomas , Øymar Knut
Published: October 2013

Aim

To assess whether inflammatory markers measured in urine and blood during acute bronchiolitis in infancy were associated with asthma, lung function, bronchial hyper‐responsiveness (BHR) and atopy at 11 years of age.

Methods

We included 105 children hospitalised for bronchiolitis during their first year of life. At hospitalisation, urinary (U‐) eosinophil protein X, U‐leukotriene E4, U‐prostaglandin 9α, 11β‐PGF2 and blood eosinophil counts were measured. Ninety‐five children (90%) were available for follow‐up at 11 years of age.

Results

At follow‐up, higher blood eosinophil counts obtained during bronchiolitis were observed in the group with asthma than in the group without asthma (median 0.27 versus 0.09 × 109/L, respectively, p = 0.048). By regression analyses, blood eosinophil counts during the acute bronchiolitis were positively associated with BHR (p = 0.006) and negatively associated with forced expiratory volume in first second (p = 0.025) at 11 years of age. None of the other inflammatory markers were associated with asthma, lung function, BHR or atopy at 11 years of age.

Conclusion

Eosinophil inflammation during bronchiolitis may have a long‐term impact on lung function and airway responsiveness. The associations could be related to virus–host interactions during bronchiolitis or to predisposed children.
Lipid profile and atherogenic indices soon after birth in Japanese preterm infants
Authors: Shoji Hiromichi , Murano Yayoi , Mori Mari , Matsunaga Nobuaki , Ohkawa Natsuki , Suganuma Hiroki , Ikeno Mitsuru , Hisata Ken , Hirayama Satoshi , Ueno Tsuyoshi , Miida Takashi , Shimizu Toshiaki
Published: October 2013

Aim

The intra‐uterine environment affects the risk of development of cardiovascular disease in adulthood. The aim of this study was to determine the influence of prematurity and foetal growth restriction on lipid metabolism, by assessing atherogenic indices soon after birth in preterm infants.

Methods

Blood samples were collected within 20 min of birth from 80 preterm infants with a gestational age of ≤35 weeks. Serum total cholesterol (TC), low‐density lipoprotein cholesterol (LDLc), high‐density lipoprotein cholesterol (HDLc), apolipoprotein‐A1 (apoA1) and apolipoprotein‐B (apoB) levels were measured. The ratio of TC/HDLc, LDLc/HDLc and apoB/apoA1 were also calculated. Correlations between these indices and gestational age, birth weight and the standard deviation (SD) score for birth weight were also determined.

Results

Gestational age, birth weight and SD score for birth weight were negatively correlated with the TC/HDLc, LDLc/HDLc and apoB/apoA1 ratios.

Conclusion

In preterm infants, prematurity and poor foetal growth may influence lipid and apolipoprotein metabolism and affect atherogenic indices at birth.
The importance of language in the home for school‐age children with permanent hearing loss
Authors: Vohr Betty R , Topol Deborah , Watson Victoria , St Pierre Lucille , Tucker Richard
Published: October 2013

Aim

To determine the characteristics of the natural spoken language environment associated with better language skills at school age of children with permanent hearing loss (HL) screened in a newborn hearing screen programme.

Methods

A language processor recorded language environment including child vocalisations (CVs), conversational turns (CTs), adult word count (AWC), percentage of language and percentage of silence in 23 children withHLand 41 hearing controls. Relationships of language environment scores withReynell language scores were analysed.

Results

Children withHLhad significantly lower Reynell comprehension scores (15–25 points; p = 0.0001) and expressive scores (7–12 points;NS) than hearing children. In regression analyses for the children withHL, after adjusting for age of entry to early intervention and stay in aNICU, every increase in ten percentage points of language in the home was associated with 7.2 points higher comprehension score (p = 0.027) and 9.99 points higher expressive score (p = 0.006). After adjusting for nonverbal intelligence, similar independent effects of the language environment on both comprehension (p = 0.027) and expressive scores (p = 0.006) were identified.

Conclusion

A rich language environment in the home is associated with better language abilities, which are necessary for academic success for children with permanentHLat school age.
Characteristics of girls with pituitary hyperplasia and sexual precocity secondary to primary hypothyroidism
Authors: Hu Yan‐Yan , Li Gui‐Mei , Hu Wen‐Wen , Wang Yi
Published: October 2013

Aim

To investigate the different characteristics of girls with pituitary hyperplasia secondary to primary hypothyroidism (PPH), with and without sexual precocity.

Methods

Pituitary hyperplasia secondary to primary hypothyroidism girls were divided into two groups: group A, with sexual precocity (n = 8), and group B, without sexual precocity (n = 14). The following values were measured before and after 3 months levothyroxine therapy: free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin (TSH), follicle‐stimulating hormone (FSH), luteinising hormone (LH), prolactin (PRL), estradiol (E2), testosterone (T), pituitary height, uterine volume (UV), ovarian volume (OV) and follicular diameter (FD).

Results

Thyrotropin, FSH, PRL, E2, T, pituitary height, UV, OV and FD were higher in group A than group B (all p < 0.05); FT3, FT4 and LH levels were not different (all p > 0.05). In all of the PPH girls, pituitary height was correlated with FT3, FT4 and PRL (r = −0.545, p = 0.009, r = −0.567, p = 0.006 and r = 0.666, p = 0.001, respectively). PRL was positively correlated with UV, FD and FSH (r = 0.581, p = 0.005, r = 0.482, p = 0.031 and r = 0.667, p = 0.001, respectively), and FSH was positively correlated with OV (r = 0.522, p = 0.013). These abnormalities regressed after therapy.

Conclusion

Pituitary hyperplasia secondary to primary hypothyroidism girls with sexual precocity have increased plasma TSH, FSH, PRL, E2 and T levels, pituitary height and uterine, ovarian and follicular size.
Aortic intima‐media thickness measured by trans‐abdominal ultrasound as an early life marker of subclinical atherosclerosis
Authors: McCloskey Kate , Vuillermin Peter , Ponsonby Anne‐Louise , Cheung Michael , Skilton Michael R , Burgner David
Published: October 2013

Atherosclerosis is a chronic inflammatory process that begins in early life. Improved identification of markers of early atherosclerosis via neonatal aortic intima‐media thickness (aIMT) measurement may allow the development of interventions to prevent or reduce later cardiovascular disease.

Conclusion

Using aIMT, studies have shown that antenatal factors such as intra‐uterine growth retardation, prematurity, maternal factors and inflammation are associated with early cardiovascular changes.
Distinguishing the impact of dexlansoprazole on heartburn vs. regurgitation in patients with gastro‐oesophageal reflux disease
Authors: Peura D. A. , Pilmer B. , Hunt B. , Mody R. , Perez M. C.
Published: October 2013

Background

Gastro‐oesophageal reflux disease (GERD) is characterised by symptomatic heartburn and regurgitation. Treatment with proton pump inhibitors (PPI) effectively decreases heartburn symptoms, but their effects on symptomatic regurgitation are less clear.

Aim

To determine the impact of PPI therapy on heartburn and regurgitation severity in patients with either non‐erosive GERD (NERD) or erosive oesophagitis (EE).

Methods

Endoscopically‐confirmed NERD patients received dexlansoprazole 30 or 60 mg or placebo in a randomised, blinded, 4‐week study. Endoscopically‐confirmed EE patients received dexlansoprazole 60 mg or lansoprazole 30 mg in two 8‐week, randomised, blinded healing studies. The Patient Assessment of Upper Gastrointestinal Symptom Severity questionnaire, which includes a heartburn/regurgitation subscale, was administered to assess symptom severity at baseline, and at weeks 2 and 4 of the NERD study and at weeks 4 and 8 during the EE trials. We defined separate subscales for heartburn and regurgitation for this post‐hoc analysis. Among patients with both symptoms at baseline, improvements in individual heartburn and regurgitation subscales along with the original combined heartburn/regurgitation subscale were determined.

Results

In the NERD and EE studies, 661 and 1909 patients, respectively, had both heartburn and regurgitation at baseline. NERD patients receiving dexlansoprazole 30 and 60 mg experienced significantly greater improvements in symptom severity for both heartburn and regurgitation compared with placebo. EE patients receiving dexlansoprazole 60 mg had significantly greater improvements in heartburn/regurgitation and heartburn‐only subscales at week 4 compared with those receiving lansoprazole.

Conclusions

Dexlansoprazole appears to be effective in improving both heartburn and regurgitation, and this improvement is maintained for the duration of treatment.
Paediatric gastroenterology evaluation of overweight and obese children referred from primary care for suspected non‐alcoholic fatty liver disease
Authors: Schwimmer J. B. , Newton K. P. , Awai H. I. , Choi L. J. , Garcia M. A. , Ellis L. L. , Vanderwall K. , Fontanesi J.
Published: October 2013

Background

Screening overweight and obese children for non‐alcoholic fatty liver disease (NAFLD) is recommended by paediatric and endocrinology societies. However, gastroenterology societies have called for more data before making a formal recommendation.

Aim

To determine whether the detection of suspected NAFLD in overweight and obese children through screening in primary care and referral to paediatric gastroenterology resulted in a correct diagnosis of NAFLD.

Methods

Information generated in the clinical evaluation of 347 children identified with suspected NAFLD through screening in primary care and referral to paediatric gastroenterology was captured prospectively. Diagnostic outcomes were reported. The diagnostic performance of two times the upper limit of normal (ULN) for alanine aminotransferase (ALT) was assessed.

Results

Non‐alcoholic fatty liver disease was diagnosed in 55% of children identified by screening and referral. Liver disease other than NAFLD was present in 18% of those referred. Autoimmune hepatitis was the most common alternative diagnosis. Children with NAFLD had significantly (P < 0.05) higher screening ALT (98 ± 95) than children with liver disease other than NAFLD (86 ± 74). Advanced fibrosis was present in 11% of children. For the diagnosis of NAFLD, screening ALT two times the clinical ULN had a sensitivity of 57% and a specificity of 71%.

Conclusions

Screening of overweight and obese children in primary care for NAFLD with referral to paediatric gastroenterology has the potential to identify clinically relevant liver pathology. Consensus is needed on how to value the risk and rewards of screening and referral, to identify children with liver disease in the most appropriate manner.
Ex vivo bupivacaine treatment results in increased adipogenesis of skeletal muscle cells in the rat
Authors: Yamanouchi Keitaro , Nakamura Katsuyuki , Takegahara Yuki , Nakano Shin‐ichi , Nishihara Masugi
Published: October 2013

Intramuscular adipose tissue (IMAT) is observed in some skeletal muscle pathologies. IMAT is implicated not only in the disorders of muscle contraction, but also of metabolism and insulin sensitivity due to its nature as a secretary organ. Several studies indicate the presence of cells with adipogenic potential in skeletal muscle. However, the mechanism of fate specification that triggers these cells to enter an adipogenic program in vivo remains to be solved. In the present study, we examined whether activation of the adipogenic program of muscle‐resident cells precedes their proliferation upon muscle injury. For this purpose, muscle injury was induced by injecting bupivacaine (BPVC) to excised skeletal muscle ex vivo. Cells isolated from ex vivo BPVC‐treated muscle exhibited higher adipogenic potential than those from saline‐treated muscle. Pre‐plating exposure of skeletal muscle cells to basic fibroblast growth factor (bFGF) mimicked the effect of ex vivo BPVC‐treatment, suggesting that bFGF released from extracellular matrix in response to muscle injury activates their adipogenic program. Interestingly, the number of myotubes were significantly reduced in the culture from BPVC‐treated muscle, suggesting that adipocytes negatively regulate myogenesis.
Vegetation succession in restoration of disturbed sites in Central Europe: the direction of succession and species richness across 19 seres
Authors: Prach Karel , Řehounková Klára , Lencová Kamila , Jírová Alena , Konvalinková Petra , Mudrák Ondřej , Študent Vojtěch , Vaněček Zdeněk , Tichý Lubomír , Petřík Petr , Šmilauer Petr , Pyšek Petr
Published: October 2013

Questions

(1) How do seres differ with respect to vegetation changes? (2) What are the directions of succession? (3) How do species numbers change? (4) How do target species, i.e. those typical of natural and semi‐natural vegetation, participate in succession? (5) Are spontaneously developed successional stages acceptable from the point of view of ecosystem restoration?

Location

Extracted peatlands, bulldozed sites in forests destroyed by air pollution, an emerged bottom of a water reservoir, corridors of former Iron Curtain, artificial fishpond islands and barriers, sedimentary basins, spoil heaps from mining, stone quarries, forest clearings, road verges, sand and gravel‐sand pits, ruderal urban sites, river gravel bars and abandoned arable fields, located in various parts of the Czech Republic in Central Europe.

Methods

Phytosociological relevés were recorded in 10–25 m2 plots located in the centre of representative successional stages defined by their age, ranging from 1 to 100 yrs. In total, we obtained 2392 vegetation samples containing 951 species. We performed DCA ordination to compare 19 seres. Desirable target species were considered as those representing (semi)‐natural vegetation and all Red List species.

Results

The seres studied are more similar in their species composition in the initial and early stages, in which synathropic species prevail, than in the later stages when the vegetation differentiates. This divergence is driven mainly by local moisture conditions. In most cases, succession led to woodland, which usually established after ca. 20 yrs. In very dry or wet places (with limited presence of woody species) open vegetation developed, often highly valuable from the restoration and conservation point of view. The total number of species and the number of target species increased in the majority of seres with successional age.

Conclusions

The vegetation in the sites studied formed a continuum along a moisture gradient and by successional age. The individual seres largely overlapped in their species composition; the sere identity was not significant. Spontaneous succession usually proceeded towards woodland, except at very dry or wet sites, and generally appeared to be an ecologically suitable way of ecosystem restoration of disturbed sites because target species became dominant over time.
Role of Pre‐Junctional CB1, But not CB2, TRPV1 or GPR55 Receptors in Anandamide‐Induced Inhibition of the Vasodepressor Sensory CGRPergic Outflow in Pithed Rats
Authors: Marichal‐Cancino Bruno A. , Altamirano‐Espinoza Alain H. , Manrique‐Maldonado Guadalupe , MaassenVanDenBrink Antoinette , Villalón Carlos M.
Published: October 2013

Stimulation of the perivascular sensory outflow in pithed rats produces vasodepressor responses mediated by CGRP release. Interestingly, endocannabinoids such as anandamide (which interacts with CB1, CB2, TRPV1 and GPR55 receptors) can regulate the activity of perivascular sensory nerves in dural blood vessels by modulating CGRP release. Yet, as no publication has reported whether this mechanism is operative in the healthy systemic vasculature, this study has specifically analysed the receptors mediating the potential inhibitory effects of the cannabinoid (CB) receptor agonists anandamide (non‐selective), JWH‐015 (CB2) and lysophosphatidylinositol (GPR55) on the rat vasodepressor sensory CGRPergic outflow (an index of systemic vasodilatation). Healthy pithed rats were pre‐treated with consecutive i.v. continuous infusions of hexamethonium, methoxamine and the above agonists. Electrical spinal (T9–T12) stimulation of the vasodepressor sensory CGRPergic outflow or i.v. injections of α–CGRP produced frequency‐dependent or dose‐dependent vasodepressor responses. The infusions of anandamide in a dose‐dependent manner inhibited the vasodepressor responses by electrical stimulation (remaining unaffected by JWH‐015 or lysophosphatidylinositol), but not those by α–CGRP. After i.v. administration of antagonists, the inhibition by 3.1 μg/kg min anandamide was: (i) potently blocked by 31–100 μg/kg NIDA41020 (CB1), (ii) unaffected by 180 μg/kg AM630 (CB2), 31 μg/kg cannabidiol (GPR55) or 31–100 μg/kg capsazepine (TRPV1) and (iii) slightly blocked by 310 μg/kg AM630. The above doses of antagonists were enough to block their respective receptors. These results suggest that anandamide‐induced inhibition of the vasodepressor sensory CGRPergic outflow is mainly mediated by pre‐junctional activation of CB1 receptors, with no pharmacological evidence for the role of CB2, TRPV1 or GPR55 receptors.
Bone marrow stroma‐mediated resistance to FLT3 inhibitors in FLT3‐ITD AML is mediated by persistent activation of extracellular regulated kinase
Authors: Yang Xiaochuan , Sexauer Amy , Levis Mark
Published: October 2013

A consistent pattern of response has been observed when FMS‐like tyrosine kinase 3 (FLT3) tyrosine kinase inhibitors (TKIs) have been used as monotherapy to treat patients with relapsed or refractory FLT3‐ internal tandem duplication (ITD) acute myeloid leukaemia (AML). Circulating blasts are cleared from the peripheral blood, while bone marrow blasts are either unaffected or are cleared from the marrow at a much slower rate. We used an in vitro model of FLT3‐ITD AML blasts co‐cultured with normal human bone marrow stromal cells to investigate the basis for this dichotomous response pattern to FLT3 inhibitors. We have found that in blasts on stroma, potent FLT3 inhibition predominantly results in cell cycle arrest rather than apoptosis. The anti‐apoptotic effect is mediated through a combination of direct cell‐cell contact and soluble factors. The addition of exogenous FLT3 ligand (FL) augments the protection, primarily by shifting the 50% inhibitory concentration for FLT3 inhibition upwards. Cytokine‐activated extracellular regulated kinase (ERK), rather than STAT5, appears to be the most important downstream signalling protein mediating the protective effect, and inhibition of MEK significantly abrogates stromal‐mediated resistance. These findings explain the phenomenon of peripheral blood versus bone marrow blast responses and suggest that the combination of potent FLT3 inhibition and MEK inhibition is a promising strategy for the treatment of FLT3‐ITD AML.
Financial Participation: Does the Risk Transfer Story Hold in France?
Authors: Baghdadi Leila , Bellakhal Rihab , Diaye Marc‐Arthur
Published: October 2013

Several articles report a positive effect of financial participation (profit sharing (PS) and employee share ownership) on firms' economic performance. This increase can be obtained in two main ways: by increasing the effort (extrinsic, intrinsic or commitment) of workers, directly or indirectly through worker selection; or by transferring more risk to the workers. The question is, of course, not neutral. Indeed, if the risk transfer story is true then it means that the increase of economic performance is obtained at the expense of workers, who take on the burden of more risks. The question is especially important in France where financial participation is associated with tax exemption for firms and where it is forbidden by law to substitute base wage and PS. The purpose of our article is to use an employer–employee dataset to answer the question of whether financial participation schemes are mainly designed as a risk transfer (from firms to workers) device.
External validation of preoperative and postoperative nomograms for prediction of cancer‐specific survival, overall survival and recurrence after robot‐assisted radical cystectomy for urothelial carcinoma of the bladder
Authors: Al‐Daghmin Ali , English Stephen , Kauffman Eric C. , Din Rakeeba , Khan Aabroo , Syed Johar R. , Sztorc Jenna , Mehedint Diana , Sharif Mohammad , Shi Yi , Wilding Gregory , Guru Khurshid A.
Published: October 2013

Objective

To externally validate currently available bladder cancer nomograms for prediction of all‐cause survival (ACS), cancer‐specific survival (CSS), other‐cause mortality (OCM) and progression‐free survival (PFS).

Patients and Methods

Retrospective analysis of a prospectively maintained database of 282 patients who underwent robot‐assisted radical cystectomy (RARC) at a single institution was performed.

The Bladder Cancer Research Consortium (BCRC), International Bladder Cancer Nomogram Consortium (IBCNC) and Lughezzani nomograms were used for external validation, and evaluation for accuracy at predicting oncological outcomes.

The 2‐ and 5‐year oncological outcomes were compared, and nomogram performance was evaluated through measurement of the concordance (c‐index) between nomogram‐derived predicted oncological outcomes and observed oncological outcomes.

Results

The median (range) patient age was 70 (36–90) years. At a mean follow‐up of 20 months, local or distant disease recurrence developed in 30% of patients. With an overall mortality rate of 33%, 17% died from bladder cancer.

The actuarial 2‐ and 5‐year PFS after RARC was 62% (95% confidence interval [CI] 54–68) and 55% (95% CI 46–63), respectively.

The actuarial 2‐ and 5‐year ACS was 66% (95% CI 59–72) and 47% (95% CI 37–55), respectively, and the 2‐ and 5‐year CSS was 81% (95% CI 74–86) and 67% (95% CI 57–76), respectively.

The PFS c‐index for IBCNC was 0.70 at 5 years, and for BCRC was 0.77 at both the 2 and 5 years. The accuracy of ACS and CSS prediction was evaluated using the BCRC and Lughezzani nomograms. Using the BCRC nomogram, c‐indices of for 2‐ and 5‐year ACS were each 0.73 and c‐indices for 2‐ and 5‐year CSS were 0.70 each. The performance of Lughezzani nomogram for 5‐year ACS, cancer‐specific mortality and OCM were 0.73, 0.72 and 0.40, respectively.

The BCRC nomogram prediction of advanced pathological stage and lymph node metastasis was modest, with c‐indices of 0.66 and 0.61, respectively.

Conclusions

Bladder cancer nomograms available from the current open RC literature adequately predict ACS, CSS and PFS after RARC.

However, prediction of advanced tumour stage and lymph node metastasis was modest and the Lughezzani nomogram failed to predict OCM.
Midkine in nephrogenesis, hypertension and kidney diseases
Authors: Sato Waichi , Sato Yuka
Published: October 2013

Midkine (MK; K; gene abbreviation, Mdk: mus musculus, MDK: homo sapiens) is a multifunctional heparin‐binding growth factor that regulates cell growth, survival and migration as well as anti‐apoptotic activity in nephrogenesis and development. Proximal tubular epithelial cells are the main sites of MK expression in the kidneys. The pathophysiological roles of MK are diverse, ranging from the development of acute kidney injury (AKI) to the progression of chronic kidney disease, often accompanied by hypertension, renal ischaemia and diabetic nephropathy. The obvious hypertension that develops in Mdk+/+ mouse models of renal ablation compared with Mdk−/− mice eventually leads to progressive renal failure, such as glomerular sclerosis and tubulointerstitial damage associated with elevated plasma angiotensin (Ang) II levels. MK is also induced in the lung endothelium by oxidative stress and subsequently up‐regulated by ACE, which hydrolyzes Ang II to induce further oxidative stress, thus accelerating MK generation; this leads to a vicious cycle of positive feedback in the MK‐Ang II pathway. Kidney–lung interactions involving positive feedback between the renin‐angiotensin system and MK might partly account for the pathogenesis of hypertension and kidney damage. MK is also involved in the pathogenesis of AKI and diabetic nephropathy through the recruitment of inflammatory cells. In contrast, MK plays a protective role against crescentic glomerulonephritis, by down‐regulating plasminogen activator inhibitor‐1. These diverse actions of MK might open up new avenues for targeted approaches to treating hypertension and various renal diseases.

Linked Articles

This article is part of a themed section on Midkine. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue‐4
Melatonin modulates microsomal PGE synthase 1 and NF‐E2‐related factor‐2‐regulated antioxidant enzyme expression in LPS‐induced murine peritoneal macrophages
Authors: Aparicio‐Soto M , Alarcón‐de‐la‐Lastra C , Cárdeno A , Sánchez‐Fidalgo S , Sanchez‐Hidalgo M
Published: October 2013

Background and Purpose

Increasing evidence demonstrates that melatonin regulates inflammatory and immune processes acting as both an activator and inhibitor of these responses. Nevertheless, the molecular mechanisms of its anti‐inflammatory action remain unclear. Here we have characterized the cellular mechanisms underlying the redox modulation of LPS‐stimulated inflammatory responses in murine peritoneal macrophages by melatonin to provide insight into its anti‐inflammatory effects.

Experimental Approach

Murine peritoneal macrophages were isolated and treated with melatonin in the presence or absence of LPS (5 μg·mL−1) for 18 h. Cell viability was determined using sulforhodamine B assay and NO production was measured using the Griess reaction. Pro‐inflammatory enzymes and transcription factors were detected by Western blotting.

Key Results

Without affecting cell viability, melatonin (12.5, 25, 50 and 100 μM) reduced the level of nitrites, inducible NOS (iNOS), COX‐2 and microsomal PGE synthase‐1 (mPGES1) protein, and p38 MAPK phosphorylation, and prevented NF‐κB translocation. Furthermore, melatonin treatment significantly increased NF‐E2‐related factor 2 (Nrf2) and haem oxygenase 1 (HO1) protein levels in murine macrophages exposed to LPS.

Conclusions and Implications

Melatonin reduced pro‐inflammatory mediators and enhanced the expression of HO1 via NF‐κB, p38 MAPK and Nrf2 cascade signalling pathways in murine macrophages. Thus, melatonin might be a promising target for diseases associated with overactivation of macrophages.
Cell death and the mitochondria: therapeutic targeting of the BCL‐2 family‐driven pathway
Authors: Roy M J , Vom A , Czabotar P E , Lessene G
Published: October 2013

The principal biological role of mitochondria is to supply energy to cells; although intriguingly, evolution has bestowed another essential function upon these cellular organelles: under physiological stress, mitochondria become the cornerstone of apoptotic cell death. Specifically, mitochondrial outer membrane permeabilization (MOMP) allows cell death factors such as cytochrome c to be released into the cytoplasm, thus inducing caspase activation and the eventual destruction of essential cellular components. Proteins of the B‐cell lymphoma 2 (BCL‐2) family control the tightly regulated pathway that causes MOMP. The equilibrium between pro‐survival and pro‐apoptotic members of the BCL‐2 family dictates the fate of cells, the homeostasis of organs and, by extension, the health of whole organisms. Dysregulation of this equilibrium is involved in a large number of diseases such as cancer, autoimmunity and neurodegenerative conditions. Modulating the activity of the BCL‐2 family of proteins with small molecules or peptides is an attractive but challenging therapeutic goal. This review highlights the latest developments in this field and provides evidence that this strategy is likely to have a positive effect on the treatment of still poorly addressed medical conditions.

Linked Articles

This article is part of a themed issue on Mitochondrial Pharmacology: Energy, Injury & Beyond. To view the other articles in this issue visit http://dx.doi.org/10.1111/bph.2014.171.issue‐8
Molecular analysis of ATP‐sensitive K+ channel subunits expressed in mouse vas deferens myocytes
Authors: Iwasa Kazuomi , Zhu Hai‐Lei , Shibata Atsushi , Maehara Yoshihiko , Teramoto Noriyoshi
Published: October 2013

Background and Purpose

ATP‐sensitive K+ (KATP) channels, which are composed of KIR6.x associated with sulphonylurea receptor (SUR) subunits, have been detected in native smooth muscle cells, but it is currently not known which of these is expressed in mouse vas deferens myocytes.

Experimental Approach

Pharmacological and electrophysiological properties of KATP channels in mouse vas deferens myocytes were investigated using patch clamp techniques. Molecular biological analyses were performed to examine the properties of these KATP channels.

Key Results

During conventional whole‐cell recording, pinacidil elicited an inward current that was suppressed by glibenclamide, a sulfonylurea agent, and by U‐37883A, a selective KIR6.1 blocker. When 0.3 mM ATP was added to the pipette solution, the peak amplitude of the pinacidil‐induced current was much smaller than that recorded in its absence. When 3 mM UDP, GDP or ADP was included in the pipette solution, an inward current was elicited after establishment of the conventional whole‐cell configuration, with potency order being UDP > GDP > ADP. These nucleoside diphosphate‐induced inward currents were suppressed by glibenclamide. MCC‐134, a SUR modulator, induced glibenclamide‐sensitive KATP currents that were similar to those induced by 100 μM pinacidil. In the cell‐attached configuration, pinacidil activated channels with a conductance similar to that of KIR6.1. Reverse transcription PCR analysis revealed the expression of KIR6.1 and SUR2B transcripts and immunohistochemical studies indicated the presence of KIR6.1 and SUR2B proteins in the myocytes.

Conclusions and Implications

Our results indicate that native KATP channels in mouse vas deferens myocytes are a heterocomplex of KIR6.1 channels and SUR2B subunits.
Naringenin inhibits the growth of Dictyostelium and MDCK‐derived cysts in a TRPP2 (polycystin‐2)‐dependent manner
Authors: Waheed A , Ludtmann M H R , Pakes N , Robery S , Kuspa A , Dinh C , Baines D , Williams R S B , Carew M A
Published: October 2013

Background and Purpose

Identifying and characterizing potential new therapeutic agents to target cell proliferation may provide improved treatments for neoplastic disorders such as cancer and polycystic diseases.

Experimental Approach

We used the simple, tractable biomedical model Dictyostelium to investigate the molecular mechanism of naringenin, a dietary flavonoid with antiproliferative and chemopreventive actions in vitro and in animal models of carcinogenesis. We then translated these results to a mammalian kidney model, Madin‐Darby canine kidney (MDCK) tubule cells, grown in culture and as cysts in a collagen matrix.

Key Results

Naringenin inhibited Dictyostelium growth, but not development. Screening of a library of random gene knockout mutants identified a mutant lacking TRPP2 (polycystin‐2) that was resistant to the effect of naringenin on growth and random cell movement. TRPP2 is a divalent transient receptor potential cation channel, where mutations in the protein give rise to type 2 autosomal dominant polycystic kidney disease (ADPKD). Naringenin inhibited MDCK cell growth and inhibited cyst growth. Knockdown of TRPP2 levels by siRNA in this model conferred partial resistance to naringenin such that cysts treated with 3 and 10 μM naringenin were larger following TRPP2 knockdown compared with controls. Naringenin did not affect chloride secretion.

Conclusions and Implications

The action of naringenin on cell growth in the phylogenetically diverse systems of Dictyostelium and mammalian kidney cells, suggests a conserved effect mediated by TRPP2 (polycystin‐2). Further studies will investigate naringenin as a potential new therapeutic agent in ADPKD.

Linked Articles

This article is part of a themed section on the pharmacology of TRP channels. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue‐10
Quality control gone wrong: mitochondria, lysosomal storage disorders and neurodegeneration
Authors: Osellame L D , Duchen M R
Published: October 2013

The eukaryotic cell possesses specialized pathways to turn over and degrade redundant proteins and organelles. Each pathway is unique and responsible for degradation of distinctive cytosolic material. The ubiquitin‐proteasome system and autophagy (chaperone‐mediated, macro, micro and organelle specific) act synergistically to maintain proteostasis. Defects in this equilibrium can be deleterious at cellular and organism level, giving rise to various disease states. Dysfunction of quality control pathways are implicated in neurodegenerative diseases and appear particularly important in Parkinson's disease and the lysosomal storage disorders. Neurodegeneration resulting from impaired degradation of ubiquitinated proteins and α‐synuclein is often accompanied by mitochondrial dysfunction. Mitochondria have evolved to control a diverse number of processes, including cellular energy production, calcium signalling and apoptosis, and like every other organelle within the cell, they must be ‘recycled.’ Failure to do so is potentially lethal as these once indispensible organelles become destructive, leaking reactive oxygen species and activating the intrinsic cell death pathway. This process is paramount in neurons which have an absolute dependence on mitochondrial oxidative phosphorylation as they cannot up‐regulate glycolysis. As such, mitochondrial bioenergetic failure can underpin neural death and neurodegenerative disease. In this review, we discuss the links between cellular quality control and neurodegenerative diseases associated with mitochondrial dysfunction, with particular attention to the emerging links between Parkinson's and Gaucher diseases in which defective quality control is a defining factor.

Linked Articles

This article is part of a themed issue on Mitochondrial Pharmacology: Energy, Injury & Beyond. To view the other articles in this issue visit http://dx.doi.org/10.1111/bph.2014.171.issue‐8
Development of pharmacological strategies for mitochondrial disorders
Authors: Kanabus M , Heales S J , Rahman S
Published: October 2013

Mitochondrial diseases are an unusually genetically and phenotypically heterogeneous group of disorders, which are extremely challenging to treat. Currently, apart from supportive therapy, there are no effective treatments for the vast majority of mitochondrial diseases. Huge scientific effort, however, is being put into understanding the mechanisms underlying mitochondrial disease pathology and developing potential treatments. To date, a variety of treatments have been evaluated by randomized clinical trials, but unfortunately, none of these has delivered breakthrough results. Increased understanding of mitochondrial pathways and the development of many animal models, some of which are accurate phenocopies of human diseases, are facilitating the discovery and evaluation of novel prospective treatments. Targeting reactive oxygen species has been a treatment of interest for many years; however, only in recent years has it been possible to direct antioxidant delivery specifically into the mitochondria. Increasing mitochondrial biogenesis, whether by pharmacological approaches, dietary manipulation or exercise therapy, is also currently an active area of research. Modulating mitochondrial dynamics and mitophagy and the mitochondrial membrane lipid milieu have also emerged as possible treatment strategies. Recent technological advances in gene therapy, including allotopic and transkingdom gene expression and mitochondrially targeted transcription activator‐like nucleases, have led to promising results in cell and animal models of mitochondrial diseases, but most of these techniques are still far from clinical application.

Linked Articles

This article is part of a themed issue on Mitochondrial Pharmacology: Energy, Injury & Beyond. To view the other articles in this issue visit http://dx.doi.org/10.1111/bph.2014.171.issue‐8