Suspected infant abuse: radiographic skeletal survey practices in pediatric health care facilities.
To determine current national radiographic skeletal survey imaging practices, including migration to digital technologies, for evaluation of suspected infant abuse. Of 155 children's health care facilities in the United States in which radiographic skeletal surveys are performed for suspected infant (<1 year old) abuse, 137 (88.4%) agreed to complete a questionnaire. Questions included facility type, imaging department volume, radiographic equipment, and details of skeletal survey imaging practices. Statistical analysis was performed with chi(2), Fisher exact, Pearson correlation, Spearman rank correlation, and Student t tests. One hundred seven completed questionnaires were returned. Forty-seven (43.9%) facilities used screen-film imaging; 60 (56.1%) used digital. Of screen-film users, 25 (53.2%) had already migrated or planned to migrate to digital within 1 year. Of screen-film users, 27 (60.0%) reported use of a high-detail imaging system, while 13 (21.7%) digital users employed a high-resolution technique (P < .001). Eighty-four (78.5%) facilities reported more than 10 images in their protocol, and 45 (42.0%) specified more than 15 images. Only one (0.9%) facility obtained fewer than three images. Upper extremities were imaged separately with at least two exposures in 81 (75.7%) facilities. Lower extremities were imaged separately with at least two exposures in 82 (76.6%) facilities. One hundred five (98.1%) facilities reported acquisition of lateral spinal views. Within U.S. pediatric health care facilities, most skeletal surveys in cases of suspected infant abuse include separate frontal views of the appendicular skeleton and frontal and lateral views of the axial skeleton. Imaging protocols and other image quality determinants vary widely, and as U.S. pediatric health care facilities migrate from film-based to digital imaging technology, imaging practices directly applicable to the digital environment are being retained; however, less attention is being paid to technical elements specific to digital imaging that affect high-detail image quality.


Functional consequences of an LMNA mutation associated with a new cardiac and non-cardiac phenotype.
Heritable dilated cardiomyopathy is a genetically highly heterogeneous disease. To date 17 different chromosomal loci have been described for autosomal dominant forms of dilated cardiomyopathy with or without additional clinical manifestations. Among the 10 mutated genes associated with dilated cardiomyopathy, the lamin A/C (LMNA) gene has been reported in forms associated with conduction-system disease with or without skeletal muscle myopathy. For the first time, we report here a French family affected with a new phenotype composed of an autosomal dominant severe dilated cardiomyopathy with conduction defects or atrial/ventricular arrhythmias, and a specific quadriceps muscle myopathy. In all previously reported cases with both cardiac and neuromuscular involvement, neuromuscular disorders preceded cardiac abnormalities. The screening of the coding sequence of the LMNA gene on all family members was performed and we identified a missense mutation (R377H) in the lamin A/C gene that cosegregated with the disease in the family. Cell transfection experiments showed that the R377H mutation leads to mislocalization of both lamin and emerin. These results were obtained in both muscular (C2C12) and non-muscular cells (COS-7). This new phenotype points out the wide spectrum of neuromuscular and cardiac manifestations associated with lamin A/C mutations, with the functional consequence of this mutation seemingly associated with a disorganization of the lamina.


Variance effects in the Bongaarts-Feeney formula.
Bongaarts and Feeney have recently proposed an adjusted total fertility rate to disentangle tempo effects from changes in the quantum of fertility. We propose an extension to the Bongaarts and Feeney formula that includes variance effects: that is, changes in the variance of the fertility schedule over time. If these variance effects are ignored, the mean age at birth and the adjusted total fertility rate are biased. We provide approximations for these biases, and we extend the TFR adjustment to fertility schedules with changing variance. We apply our method to the Swedish baby boom and bust, and show that variance effects are important for evaluating the relative contributions of tempo and quantum effects to the fertility change from 1985 to 1995.


Novel operative anoscope for Ferguson hemorrhoidectomy: a feasibility study and comparative cohort analysis.
Operative hemorrhoidectomy can result in pain and altered continence from excessive excision of anoderm or surrounding tissue. We assessed a novel low-profile slotted anoscope to determine if the device would promote safe dissection, lessen trauma, and reduce operative times for hemorrhoidectomy. Patients requiring hemorrhoidectomy (June 2008 - January 2010) underwent a prospective phase-2 trial evaluating a new operating anoscope (CAD, Ethicon Endosurgery, Cincinnati, OH). Demographics and perioperative end points including bleeding, pain, fecal incontinence, stenosis, and symptom recurrence were analyzed at 4 weeks, 3 months, 6 months, and 1 year postoperatively. We compared these to patients undergoing hemorrhoidectomy (February 2010 - November 2012) with a traditional Hill-Ferguson anoscope (THF). 40 patients (CAD, 20 vs THF, 20) were included. Presenting symptoms were similar, whereas mean duration of symptoms was longer for CAD (41.2 ± 8.4 vs 27 ± 9.5 months; P < .05). Estimated blood loss was lower for CAD [8.3 mL (range = 2-40 mL) vs 11.3 mL THF (range = 5-35 mL; P = .87)]. Mean operative times were lower for the CAD than the THF group (15.6 ± 3.4 vs 26.1 ± 4.1 minutes; P < .05). Visual analog pain scores were non-significantly increased in the THF group at 4 weeks (P = .23). At 3 months, 6 months, and 1 year, there was no difference in continence. The CAD anoscope reduced operative times for modified Ferguson (closed) hemorrhoidectomy when compared with traditional retractors. There was no difference in incontinence or pain between groups.


Millisecond-timescale optical control of neural dynamics in the nonhuman primate brain.
To understand how brain states and behaviors are generated by neural circuits, it would be useful to be able to perturb precisely the activity of specific cell types and pathways in the nonhuman primate nervous system. We used lentivirus to target the light-activated cation channel channelrhodopsin-2 (ChR2) specifically to excitatory neurons of the macaque frontal cortex. Using a laser-coupled optical fiber in conjunction with a recording microelectrode, we showed that activation of excitatory neurons resulted in well-timed excitatory and suppressive influences on neocortical neural networks. ChR2 was safely expressed, and could mediate optical neuromodulation, in primate neocortex over many months. These findings highlight a methodology for investigating the causal role of specific cell types in nonhuman primate neural computation, cognition, and behavior, and open up the possibility of a new generation of ultraprecise neurological and psychiatric therapeutics via cell-type-specific optical neural control prosthetics.


Potentiation of antimicrobial activity of ciprofloxacin by Pelargonium graveolens essential oil against selected uropathogens.
The recent approach of using herbs and antibiotics in combination constitutes a strategy to overcome the problems of resistance and side effects associated with conventional antibiotics. In the present study, the antimicrobial effect of Pelargonium graveolens L' Hér essential oil in combination with ciprofloxacin was evaluated on uropathogens, namely, Klebsiella pneumoniae KT2, Proteus mirabilis PRT3 and Staphylococcus aureus ST2. Minimum inhibitory concentrations of P. graveolens essential oil and ciprofloxacin were determined by the microbroth dilution method and further, the interaction between these two agents was studied by a checkerboard method. The fractional inhibitory concentration index (FICI) was calculated to be 0.375 for both K. pneumoniae KT2 and P. mirabilis PRT3, while for S. aureus ST2 it was found to be 0.5. The values of FICI for the tested microorganisms were found to be ≤0.5, which indicates synergism between P. graveolens essential oil and ciprofloxacin. The concave shaped curve in the isobolograms also depicted a synergistic effect of P. graveolens essential oil and ciprofloxacin against the tested microorganisms. Hence, the synergistic action of P. graveolens essential oil and ciprofloxacin may be applied for the treatment of UTIs, which have hitherto been treated by using only synthetic drugs.


Effect of the number of biopsies on the subsequent acquisition of new human papillomavirus infections.
To analyze the effect of number of biopsies taken at enrollment with incident human papillomavirus (HPV) infections detected at the next 6-month visit. Using data from the Atypical Squamous Cells of Undetermined Significance (ASCUS) and Low-Grade Squamous Intraepithelial Lesion (LSIL) Triage Study (ALTS), we compared the 6-month acquisition of new HPV infections among 988 women who underwent colposcopy, were not diagnosed and treated for cervical intraepithelial neoplasia grade 2 or more severe lesions, and had polymerase chain reaction results for 38 HPV genotypes at enrollment and follow-up. Our analysis considered each woman's possible acquisition of each of these HPV genotypes. The average 6-month acquisition of any HPV genotype for women with zero, one, and two or more biopsies was 1.82%, 1.74%, and 1.97%, respectively (Ptrend=.7). In a logistic regression model that controlled for age, baseline HPV status, and having a new sexual partner during the 6-month follow-up, two or more biopsies (compared with one biopsy) was not associated with acquiring HPV (odds ratio 1.0, 95% confidence interval 0.75-1.3). Multiple biopsies compared with a single biopsy did not increase the likelihood of acquiring new HPV infections. II.


Changes in cardiac biomarkers during doxorubicin treatment of pediatric patients with high-risk acute lymphoblastic leukemia: associations with long-term echocardiographic outcomes.
Doxorubicin causes cardiac injury and cardiomyopathy in children with acute lymphoblastic leukemia (ALL). Measuring biomarkers during therapy might help individualize treatment by immediately identifying cardiac injury and cardiomyopathy. Children with high-risk ALL were randomly assigned to receive doxorubicin alone (n = 100; 75 analyzed) or doxorubicin with dexrazoxane (n = 105; 81 analyzed). Echocardiograms and serial serum measurements of cardiac troponin T (cTnT; cardiac injury biomarker), N-terminal pro-brain natriuretic peptide (NT-proBNP; cardiomyopathy biomarker), and high-sensitivity C-reactive protein (hsCRP; inflammatory biomarker) were obtained before, during, and after treatment. cTnT levels were increased in 12% of children in the doxorubicin group and in 13% of the doxorubicin-dexrazoxane group before treatment but in 47% and 13%, respectively, after treatment (P = .005). NT-proBNP levels were increased in 89% of children in the doxorubicin group and in 92% of children in the doxorubicin-dexrazoxane group before treatment but in only 48% and 20%, respectively, after treatment (P = .07). The percentage of children with increased hsCRP levels did not differ between groups at any time. In the first 90 days of treatment, detectable increases in cTnT were associated with abnormally reduced left ventricular (LV) mass and LV end-diastolic posterior wall thickness 4 years later (P < .01); increases in NT-proBNP were related to an abnormal LV thickness-to-dimension ratio, suggesting LV remodeling, 4 years later (P = .01). Increases in hsCRP were not associated with any echocardiographic variables. cTnT and NT-proBNP may hold promise as biomarkers of cardiotoxicity in children with high-risk ALL. Definitive validation studies are required to fully establish their range of clinical utility.


Fructose-1,6-bisphosphate couples glycolytic flux to activation of Ras.
Yeast and cancer cells share the unusual characteristic of favoring fermentation of sugar over respiration. We now reveal an evolutionary conserved mechanism linking fermentation to activation of Ras, a major regulator of cell proliferation in yeast and mammalian cells, and prime proto-oncogene product. A yeast mutant (tps1∆) with overactive influx of glucose into glycolysis and hyperaccumulation of Fru1,6bisP, shows hyperactivation of Ras, which causes its glucose growth defect by triggering apoptosis. Fru1,6bisP is a potent activator of Ras in permeabilized yeast cells, likely acting through Cdc25. As in yeast, glucose triggers activation of Ras and its downstream targets MEK and ERK in mammalian cells. Biolayer interferometry measurements show that physiological concentrations of Fru1,6bisP stimulate dissociation of the pure Sos1/H-Ras complex. Thermal shift assay confirms direct binding to Sos1, the mammalian ortholog of Cdc25. Our results suggest that the Warburg effect creates a vicious cycle through Fru1,6bisP activation of Ras, by which enhanced fermentation stimulates oncogenic potency.Yeast and cancer cells both favor sugar fermentation in aerobic conditions. Here the authors describe a conserved mechanism from yeast to mammals where the glycolysis intermediate fructose-1,6-bisphosphate binds Cdc25/Sos1 and couples increased glycolytic flux to increased Ras proto-oncoprotein activity.


Physiological response curve analysis using nonlinear mixed models.
Nonlinear response curves are often used to model the physiological responses of plants. These models are preferable to polynomials because the coefficients fit to the curves have biological meaning. The response curves are often generated by repeated measurements on one subject, over a range of values for the environmental variable of interest. However, the typical analysis of differences in coefficients between experimental groups does not include a repeated measures approach. This may lead to inappropriate estimation of error terms. Here, we show how to combine mixed model analysis, available in SAS, that allows for repeated observations on the same experimental unit, with nonlinear response curves. We illustrate the use of this nonlinear mixed model with a study in which two plant species were grown under contrasting light environments. We recorded light levels and net photosynthetic response on anywhere from 8 to 10 points per plant and fit a Mitscherlich model in which each plant has its own coefficients. The coefficients for the photosynthetic light-response curve for each plant were assumed to follow a multivariate normal distribution in which the mean was determined by the treatment. The approach yielded biologically relevant coefficients and unbiased standard error estimates for multiple treatment comparisons.


An analysis of growth, differentiation and apoptosis genes with risk of renal cancer.
We conducted a case-control study of renal cancer (987 cases and 1298 controls) in Central and Eastern Europe and analyzed genomic DNA for 319 tagging single-nucleotide polymorphisms (SNPs) in 21 genes involved in cellular growth, differentiation and apoptosis using an Illumina Oligo Pool All (OPA). A haplotype-based method (sliding window analysis of consecutive SNPs) was used to identify chromosome regions of interest that remained significant at a false discovery rate of 10%. Subsequently, risk estimates were generated for regions with a high level of signal and individual SNPs by unconditional logistic regression adjusting for age, gender and study center. Three regions containing genes associated with renal cancer were identified: caspase 1/5/4/12(CASP 1/5/4/12), epidermal growth factor receptor (EGFR), and insulin-like growth factor binding protein-3 (IGFBP3). We observed that individuals with CASP1/5/4/12 haplotype (spanning area upstream of CASP1 through exon 2 of CASP5) GGGCTCAGT were at higher risk of renal cancer compared to individuals with the most common haplotype (OR:1.40, 95% CI:1.10-1.78, p-value = 0.007). Analysis of EGFR revealed three strong signals within intron 1, particularly a region centered around rs759158 with a global p = 0.006 (GGG: OR:1.26, 95% CI:1.04-1.53 and ATG: OR:1.55, 95% CI:1.14-2.11). A region in IGFBP3 was also associated with increased risk (global p = 0.04). In addition, the number of statistically significant (p-value<0.05) SNP associations observed within these three genes was higher than would be expected by chance on a gene level. To our knowledge, this is the first study to evaluate these genes in relation to renal cancer and there is need to replicate and extend our findings. The specific regions associated with risk may have particular relevance for gene function and/or carcinogenesis. In conclusion, our evaluation has identified common genetic variants in CASP1, CASP5, EGFR, and IGFBP3 that could be associated with renal cancer risk.


Soy milk intake and plasma sex hormones: a cross-sectional study in pre- and postmenopausal women (EPIC-Oxford).
Soy beans contain high levels of the isoflavones genistein and daidzein and their glucosides. We investigated the relationship between soy milk intake and plasma concentrations of estradiol, sex hormone-binding globulin, and, in premenopausal women, follicle-stimulating hormone, luteinizing hormone, and progesterone in a cross-sectional study of 636 premenopausal and 456 postmenopausal British women. Sixty-five percent of the women were vegetarians or vegans. Data on soy milk intake and other factors were obtained from a validated food frequency questionnaire, hormone concentrations were measured by immunoassays, and variations in geometric means were compared using analysis of covariance. We observed no statistically significant trends or meaningful associations between soy milk intake and circulating sex hormones. Adjusting for factors possibly affecting circulating hormone concentrations did not materially alter the results. We conclude that soy milk intake does not change plasma concentrations of sex hormones in pre- or postmenopausal British women who consume soy milk as a part of their regular diet.


Combined MEK and BCL-2/XL Inhibition Is Effective in High-Grade Serous Ovarian Cancer Patient-Derived Xenograft Models and BIM Levels Are Predictive of Responsiveness.
Most patients with late-stage high-grade serous ovarian cancer (HGSOC) initially respond to chemotherapy but inevitably relapse and develop resistance, highlighting the need for novel therapies to improve patient outcomes. The MEK/ERK pathway is activated in a large subset of HGSOC, making it an attractive therapeutic target. Here, we systematically evaluated the extent of MEK/ERK pathway activation and efficacy of pathway inhibition in a large panel of well-annotated HGSOC patient-derived xenograft models. The vast majority of models were nonresponsive to the MEK inhibitor cobimetinib (GDC-0973) despite effective pathway inhibition. Proteomic analyses of adaptive responses to GDC-0973 revealed that GDC-0973 upregulated the proapoptotic protein BIM, thus priming the cells for apoptosis regulated by BCL2-family proteins. Indeed, combination of both MEK inhibitor and dual BCL-2/XL inhibitor (ABT-263) significantly reduced cell number, increased cell death, and displayed synergy in vitro in most models. In vivo, GDC-0973 and ABT-263 combination was well tolerated and resulted in greater tumor growth inhibition than single agents. Detailed proteomic and correlation analyses identified two subsets of responsive models-those with high BIM at baseline that was increased with MEK inhibition and those with low basal BIM and high pERK levels. Models with low BIM and low pERK were nonresponsive. Our findings demonstrate that combined MEK and BCL-2/XL inhibition has therapeutic activity in HGSOC models and provide a mechanistic rationale for the clinical evaluation of this drug combination as well as the assessment of the extent to which BIM and/or pERK levels predict drug combination effectiveness in chemoresistant HGSOC.


[Remote results of open interventions in treatment of abdominal aortic aneurysms].
Analysed herein are the results of treating a total of 471 patients operated on at the Department of Surgical Treatment for Arterial Pathology of the Research Centre of Cardiovascular Surgery named after A. N. Bakulev under the Russian Academy of Medical Sciences for an aneurysm of the infrarenal portion of the abdominal aorta. Complicated forms of AAA were observed in 71 (15.1%) patients. Of these, 13 patients were admitted presenting with a clinical picture of aortic rupture, 56 patients with aortic wall tear (according to ultrasound and/or MSCH findings), and two patients had break of the inferior vena cava. The most frequently encountered accompanying diseases were as follows: CAD noted in 367 (78.2%) patients. Valvular heart defects were revealed in 68 (14.4%) patients. Atherosclerotic alterations of the ascending aorta were diagnosed in 97 (20.6%) patients, and an ascending aortic aneurysm in 14 (2.8%) patients. Operations on the heart were performed in 45 (19.1%) patients. Mitral valve plasty was carried out in 7 patients, and 11 patients had a history of prior interventions on the aortic root and ascending aorta (Bentall-De Bono operation in 3 patients, Carbol operation in 3 patients). 32 patients underwent reconstructive operations on coronary arteries. Transmyocardial laser revascularization of the myocardium was carried out in 4 patients. Endovascular interventions were performed in 32 (6.8%) patients (stenting or transluminal balloon angioplasty of coronary arteries). All patients underwent AAA resection with prosthetic repair. The mentioned operations were combined with the following simultaneous interventions: aortocoronary bypass grafting performed in 31 cases, carotid endarterectomy in 27 cases, and plasty of renal arteries was carried out in 48 patients. Complications in the immediate postoperative period were observed in 79 (16.8%) patients. Hospital mortality amounted to 11 (2.3%) patients. The cumulative survival rate after 1, 5 and 10 years after surgery amounted to 93%, 81% and 44%, respectively, in a complicated course amounting to 91%, 79% and 16%, respectively. The obtained findings of the study demonstrated that the results of the AAA resection should be regarded as fairly good. The main factors influencing the quality of life and surgical outcomes in abdominal aortic aneurysms are as follows: CAD, arterial hypertension, and chronic cerebrovascular diseases. In patients without accompanying diseases, the criteria of quality of life after surgery approximate to those in a healthy population.


Manualized group cognitive behavioral therapy for social anxiety in first-episode psychosis: a randomized controlled trial.
Social anxiety (SA), a prevalent comorbid condition in psychotic disorders with a negative impact on functioning, requires adequate intervention relatively early. Using a randomized controlled trial, we tested the efficacy of a group cognitive-behavioral therapy intervention for SA (CBT-SA) that we developed for youth who experienced the first episode of psychosis (FEP). For our primary outcome, we hypothesized that compared to the active control of group cognitive remediation (CR), the CBT-SA group would show a reduction in SA that would be maintained at 3- and 6-month follow-ups. For secondary outcomes, it was hypothesized that the CBT-SA group would show a reduction of positive and negative symptoms and improvements in recovery and functioning. Ninety-six patients with an FEP and SA, recruited from five different FEP programs in the Montreal area, were randomized to 13 weekly group sessions of either CBT-SA or CR intervention. Linear mixed models revealed that multiple measures of SA significantly reduced over time, but with no significant group differences. Positive and negative symptoms, as well as functioning improved over time, with negative symptoms and functioning exhibiting a greater reduction in the CBT-SA group. While SA decreased over time with both interventions, a positive effect of the CBT-SA intervention on measures of negative symptoms, functioning, and self-reported recovery at follow-up suggests that our intervention had a positive effect that extended beyond symptoms specific to SA.ClinicalTrials.gov identifier: NCT02294409.


Regulation of cytosolic calcium by angiotensins in vascular smooth muscle.
The carboxy terminal homologue of angiotensin II (Ang II), Ang-(3-8) or hexapeptide, was used as a model peptide to examine the types of receptor mechanisms involved in calcium mobilization in cultured vascular smooth muscle cells. Hexapeptide did not produce tachyphylaxis but did produce a sustained increase in intracellular calcium. Differences in the increase in intracellular calcium [( Ca2+]i) and the pattern of inositol phosphate production indicate that Ang-(3-8) and maximal concentrations of Ang II mobilize calcium through different mechanisms. The calcium-mobilizing mechanisms that predominate appear to depend on the concentration of angiotensin. Concentrations of Ang II greater than 10(-8) M produce sharp calcium transients in which the [Ca2+]i returns close to baseline within 1 minute after stimulation, but concentrations of Ang II equal to or less than 3 x 10(-9) M result in a plateau increase in calcium. Pretreatment with Bordetella pertussis toxin does not abolish either the calcium transient induced by Ang II or the plateau phase induced by Ang-(3-8), indicating that the GTP-transducing protein that couples the receptor to phospholipase C or, possibly, a receptor-operated calcium channel is not Bordetella pertussis toxin sensitive.


A tunable multivariable nonlinear robust observer for biological systems.
This paper presents a robust nonlinear asymptotic observer with adjustable convergence rate with a great potential of applicability for biological systems in which the main state variables are difficult and expensive to measure or such measurements do not exist. This observer scheme is based on the classical asymptotic observer, which is modified to allow the tuning of the convergence rate. It is shown that the proposed observer provides fast and satisfactory estimates when facing load disturbances, system failures and parameter uncertainty while maintaining the excellent robustness and stability properties of the classical asymptotic observer. The implementation of the tunable observer is carried out by numerical simulations of a mathematical model of an anaerobic digestion process used for wastewater treatment. The key results are examined and further developed.


Laboratory evaluation of fipronil, a phenylpyrazole insecticide, against adult Anopheles (Diptera: Culicidae) and investigation of its possible cross-resistance with dieldrin in Anopheles stephensi.
Adult mosquitoes from two strains of Anopheles gambiae and from three strains of Anopheles stephensi were exposed to 0.25% fipronil-treated papers in WHO test kits or to 500 mg fipronil m-2 impregnated mosquito netting in bioassay spheres. For comparison, tests were also carried out with the pyrethroid permethrin, using the same methods and doses, and on papers treated with 0.4 and 4% of the cyclodiene insecticide dieldrin. Compared with the same doses of permethrin, fipronil showed less and delayed activity. Two of the An stephensi strains were resistant to fipronil and dieldrin. To investigate whether this was due to a resistance mechanism in the An stephensi strains acting against both insecticides, the most fipronil- and dieldrin-tolerant strain was further selected in two separate lines with one of the insecticides, followed by tests with the insecticide that the line had not been selected with. This indicated a concomitant rise of resistance to dieldrin in the fipronil-selected line and vice versa. Repeated back-crossing of the two lines with a susceptible strain and re-selection with either dieldrin or fipronil gave evidence for the involvement of a single resistance mechanisms to both insecticides. Permethrin resistance in both lines declined with selection for dieldrin or fipronil and confirms the absence of cross-resistance between fipronil and pyrethroids.


The Effects of Intraoperative Caffeine on Postoperative Opioid Consumption and Related Outcomes After Laparoscopic Surgery: A Randomized Controlled Trial.
Surgical patients are vulnerable to opioid dependency and related risks. Clinical-translational data suggest that caffeine may enhance postoperative analgesia. This trial tested the hypothesis that intraoperative caffeine would reduce postoperative opioid consumption. The secondary objective was to assess whether caffeine improves neuropsychological recovery postoperatively. This was a single-center, randomized, placebo-controlled trial. Participants, clinicians, research teams, and data analysts were all blinded to the intervention. Adult (≥18 years old) surgical patients (n = 65) presenting for laparoscopic colorectal and gastrointestinal surgery were randomized to an intravenous caffeine citrate infusion (200 mg) or dextrose 5% in water (40 mL) during surgical closure. The primary outcome was cumulative opioid consumption through postoperative day 3. Secondary outcomes included subjective pain reporting, observer-reported pain, delirium, Trail Making Test performance, depression and anxiety screens, and affect scores. Adverse events were reported, and hemodynamic profiles were also compared between the groups. Sixty patients were included in the final analysis, with 30 randomized to each group. The median (interquartile range) cumulative opioid consumption (oral morphine equivalents, milligrams) was 77 mg (33-182 mg) for caffeine and 51 mg (15-117 mg) for placebo (estimated difference, 55 mg; 95% confidence interval [CI], -9 to 118; P = .092). After post hoc adjustment for baseline imbalances, caffeine was associated with increased opioid consumption (87 mg; 95% CI, 26-148; P = .005). There were otherwise no differences in prespecified pain or neuropsychological outcomes between the groups. No major adverse events were reported in relation to caffeine, and no major hemodynamic perturbations were observed with caffeine administration. Caffeine appears unlikely to reduce early postoperative opioid consumption. Caffeine otherwise appears well tolerated during anesthetic emergence.


Evaluation of the impact of a nutritional program for undernourished children in Brazil.
To assess the effectiveness on child growth and body composition of a supplementary feeding program (Milk Supplement Program), a prospective, controlled study was conducted in Northeast Brazil. When entering the Program, children from 10 municipalities with the highest coverage rates in the Program (intervention group) were compared to non-beneficiary children from 10 municipalities with the lowest coverage rates (control group). A total of 219 children aged 6-18 months were enrolled. At entry, both groups were comparable in terms of age, sex, and nutritional status. There were frequent gaps in delivery of the supplement, no extra milk was provided to siblings less than 5 years of age, intra-household redistribution of milk was high, and maternal compliance with recommendations was low. Adjusted analyses by multilevel modelling showed average changes in weight, length, weight-age and length-age Z-scores, and % body water (deuterium method), at 6 months, of 1.53 kg, 6.34 cm, 0.33, 0.05, and 1.11% respectively among supplemented children as compared to 1.54 kg, 6.5 cm, 0.26, 0.07, and 4.10% among controls, with no statistically significant difference between groups. Thus, the Program failed to compensate for nutritional deficiencies in undernourished children in Northeast Brazil.


Biomimetic aerobic oxidative hydroxylation of arylboronic acids to phenols catalysed by a flavin derivative.
Flavin-catalysed oxidative hydroxylation of substituted arylboronic acids by molecular oxygen with the assistance of hydrazine or ascorbic acid resulted in phenols in high yields. This mild organocatalytic protocol is compatible with a variety of functional groups and it is alternatively usable for transformation of alkylboronic acids to alcohols. Reaction takes place also in water and fulfils criteria for a green procedure.


Evolutionary relationships among the protostrongylidae (Nematoda: Metastrongyloidea) as inferred from morphological characters, with consideration of parasite-host coevolution.
The phylogeny of nematodes in the family Protostrongylidae (Nematoda: Metastrongyloidea) was reconstructed by cladistic analysis of 28 binary and multistate characters derived from comparative morphology. Analyses were hierarchical, and examined (1) relationships among genera, including 13 ingroup taxa and Metastrongylidae as an outgroup (single tree, 78 steps, consistency index [CI] = 0.705); and (2) relationships among genera and species groups, including 21 ingroup taxa and Metastrongylus apri as an outgroup (single tree, 76 steps, CI = 0.582). In the species-level tree, Protostrongylidae was divided into 2 major clades, 1 containing the subfamilies Muelleriinae (including the recently described Umingmtakstrangylus pallikuukensis), Elaphostrongylinae, and the Varestrongylinae (excluding Pneumocaulus kadenazii). Varestrongylus was paraphyletic as it included Pneumostrongylus calcaratus. The second major clade consisted of a paraphyletic group containing Protostrongylus spp. and Spiculocaulus leuckarti and, basal to this subclade, several other individual protostrongylid lineages. The various subclades generally correspond to the subfamilial divisions of the Protostrongylidae. The Neostrongylinae, however, is not supported as Neostrongylus and Orthostrongylus are not sister groups. Based on a large number of hypothesized synapomorphies, the elaphostrongylines appear to be a highly derived group of protostrongylids, a feature potentially correlated with their habitat localization in muscular and nervous tissues. The generic-level tree retained most of the primary structure revealed among the species but excluded the varestrongylines from the Muelleriinae + Elaphostrongylinae subclade. Artiodactyles of the family Cervidae are considered basal hosts for protostrongylids; secondary colonization in Caprini, Rupicaprini, and among lagomorphs is postulated.


On an early gene for membrane-integral inorganic pyrophosphatase in the genome of an apparently pre-luca extremophile, the archaeon Candidatus Korarchaeum cryptofilum.
A gene for membrane-integral inorganic pyrophosphatase (miPPase) was found in the composite genome of the extremophile archaeon Candidatus Korarchaeum cryptofilum (CKc). This korarchaeal genome shows unusual partial similarity to both major archaeal phyla Crenarchaeota and Euryarchaeota. Thus this Korarchaeote might have retained features that represent an ancestral archaeal form, existing before the occurrence of the evolutionary bifurcation into Crenarchaeota and Euryarchaeota. In addition, CKc lacks five genes that are common to early genomes at the LUCA border. These two properties independently suggest a pre-LUCA evolutionary position of this extremophile. Our finding of the miPPase gene in the CKc genome points to a role for the enzyme in the energy conversion of this very early archaeon. The structural features of its miPPase indicate that it can pump protons through membranes. An miPPase from the extremophile bacterium Caldicellulosiruptor saccharolyticus also has a sequence indicating a proton pump. Recent analysis of the three-dimensional structure of the miPPase from Vigna radiata has resulted in the recognition of a strongly acidic substrate (orthophosphate: Pi, pyrophosphate: PPi) binding pocket, containing 11 Asp and one Glu residues. Asp (aspartic acid) is an evolutionarily very early proteinaceous amino acid as compared to the later appearing Glu (glutamic acid). All the Asp residues are conserved in the miPPase of CKc, V. radiata and other miPPases. The high proportion of Asp, as compared to Glu, seems to strengthen our argument that biological energy conversion with binding and activities of orthophosphate (Pi) and energy-rich pyrophosphate (PPi) in connection with the origin and early evolution of life may have started with similar or even more primitive acidic peptide funnels and/or pockets.


Dynamics of aquaporin-2 serine-261 phosphorylation in response to short-term vasopressin treatment in collecting duct.
We recently identified a novel phosphorylation site, serine-261 (pS261), in the COOH-terminus of the vasopressin-regulated water channel, aquaporin-2 (AQP2). To address whether phosphorylation at this site is regulated by vasopressin, a rabbit polyclonal phospho-specific antibody was generated. Dot blot and immunoblot analysis demonstrated that this antibody specifically recognizes AQP2 phosphorylated at pS261, and that phosphorylation of S256 (pS256), a site already known to be regulated by vasopressin, does not interfere with antibody recognition. Immunohistochemical analysis revealed intense pS261 labeling of inner medullary collecting duct (IMCD) from wild-type mice, while sections from AQP2 knockout animals showed a general absence of labeling. AQP2 pS261 was present in principal cells of all mouse and rat distal tubule segments from the connecting tubule to the terminal IMCD. Co-immunolabeling of collecting duct with phospho-specific and total AQP2 antibodies revealed that pS261 and pS256 have distinct subcellular distributions. Levels of pS256 increased, while the amount of pS261 significantly decreased in freshly isolated rat IMCD samples incubated with 1 nM [deamino-Cys(1),D-Arg(8)]vasopressin for 30 min. Similarly, based on immunohistochemical labeling, the amount of pS261 was reduced in all collecting duct segments of Brattleboro rats treated with [deamino-Cys(1),D-Arg(8)]vasopressin for 2 h. This study reveals a reciprocal change in S256 and S261 phosphorylation in response to short-term vasopressin exposure, suggesting that these residues may serve distinct roles in regulation of AQP2 subcellular distribution and collecting duct water permeability.


Efficiency of adjuvant immunochemotherapy following curative resection in patients with locally advanced gastric cancer.
Despite curative resection, 50%-90% of gastric cancer patients die of disease relapse. Although some clinical trials have indicated that chemotherapy and immunochemotherapy may be effective modalities, more recent studies have not been able to define the standard treatment for advanced gastric cancer. The present study evaluated the effect of adjuvant immunochemotherapy with the use of BCG (bacille Calmette-Guerin) and FAM (5-fluorouracil, adriamycin, mitomycin C) chemotherapy on the survival of patients with locally advanced resectable gastric cancer. A total of 156 patients with stage III or IV gastric cancer who had undergone curative resection were randomly assigned to three treatment groups: BCG + FAM (immunochemotherapy), FAM (chemotherapy), and control (surgery only). Treatment was continued for 2 years or until death. Further postsurgical follow up was carried on for up to 10 years. Overall 10-year survival was 47.1% for the immunochemotherapy group (P < 0.037 vs FAM and P < 0.0006 vs control), 30% for the chemotherapy group (vs control, NS), and 15.2% for the control group. In patients with pT2/T3 primary tumors, 10-year survival was 55.3% for BCG + FAM vs 28.2% for FAM (P < 0.01) and 14.6% for the control group (P < 0.00018). BCG + FAM significantly improved the survival of patients with intestinal-type but not diffuse-type cancer. Immunochemotherapy was well tolerated. This study, based on a limited number of patients, indicates that adjuvant immunochemotherapy (BCG + FAM) may prolong the survival of gastric cancer patients after curative gastrectomy; in particular, in patients with pT2/T3 tumors and intestinal-type primary tumors. There was no survival benefit from FAM adjuvant chemotherapy.


Idiopathic granulomatous mastitis: dilemmas in diagnosis and treatment.
Idiopathic granulomatous mastitis (IGM) is a benign rare inflammatory disease of the breast. Due to its uncommon etiology, diagnosis and treatment is still unknown. Selection of a standard method for diagnosing idiopathic granulomatous mastitis is sophisticated. In view of non-definitive clinical and imaging finding, histopathology is the cornerstone of definitive diagnosis. To determine and help solve the dilemma of treatment and diagnosis of idiopathic granulomatous mastitis. This historical cohort study was conducted on 48 patients who referred to the general surgery clinic of Imam Khomeini Hospital of Urmia, were diagnosed with IGM and were histopathologically selected by census using the registry system, in Urmia city, Iran, during 2010-2015 so that medical reports, ultrasonography (US) and mammography (MMG) findings, follow-up information and recurrence rate were obtained from records. The data were analyzed using SPSS software version 18 and descriptive statistics were used. According to records, 68.75% of patients (n=33) had palpable mass, 45.83% of patients (n=22) had breast pain and swelling, erythema and 20.83% of patients (n=10) had purulent drainage. Of the 48 patients 12 (25%) had mammography, which revealed the following findings: mass with irregular border in 6 patients, skin thickness in 2 cases (4.16%), and parenchymal asymmetry in 4 cases (8.33%). Minimum follow-up was 24 (range 24-56) months. According to our findings, histopathology of the disease is fundamental for correct diagnosis. Steroid therapy as a therapeutic method such as prednisolone was an effective and applicable choice in the treatment of idiopathic granulomatous mastitis by decreasing in inflammation.


Talent Identification in an English Premiership Rugby Union Academy: Multidisciplinary Characteristics of Selected and Non-selected Male Under-15 Players.
Entry into an academy can be a defining moment for a promising young player. The aim of this study was to explore the multidimensional characteristics that differentiated selected and non-selected male under-15 rugby union players at an English Premiership academy. Seventy-four players (mean age 14.6 ± 0.3 years: selected n = 29; non-selected n = 45) were measured across nine characteristics from four overarching factors: (a) anthropometric (n = 2), (b) physiological (n = 5), (c) cognitive (n = 1), and (d) birth quartile. An ANOVA compared differences between groups (selected vs. non-selected), whilst a Welch's t-test and Cohen's d were used for further comparisons. A multivariate logistic regression was also used to predict selection. Results showed significant differences between selected and non-selected players for anthropometric (P = 0.021) and physiological factors (P < 0.001). Moreover, relatively older players were overrepresented with 65% born in the first half of the year, whereas no significant differences were apparent for the cognitive test. More specifically, selected players possessed greater body mass (P = 0.022, d = 0.5) and handgrip strength (P = 0.020, d = 0.5) compared to non-selected players, whilst multivariate analysis showed the 20 m sprint explained 25.4% of the variance (P = 0.001). Overall, it appears selection into an English Premiership rugby union academy may be due to enhanced physical attributes rather than cognitive abilities.


Pax-2 controls multiple steps of urogenital development.
Urogenital system development in mammals requires the coordinated differentiation of two distinct tissues, the ductal epithelium and the nephrogenic mesenchyme, both derived from the intermediate mesoderm of the early embryo. The former give rise to the genital tracts, ureters and kidney collecting duct system, whereas mesenchymal components undergo epithelial transformation to form nephrons in both the mesonephric (embryonic) and metanephric (definitive) kidney. Pax-2 is a transcriptional regulator of the paired-box family and is widely expressed during the development of both ductal and mesenchymal components of the urogenital system. We report here that Pax-2 homozygous mutant newborn mice lack kidneys, ureters and genital tracts. We attribute these defects to dysgenesis of both ductal and mesenchymal components of the developing urogenital system. The Wolffian and Müllerian ducts, precursors of male and female genital tracts, respectively, develop only partially and degenerate during embryogenesis. The ureters, inducers of the metanephros are absent and therefore kidney development does not take place. Mesenchyme of the nephrogenic cord fails to undergo epithelial transformation and is not able to form tubules in the mesonephros. In addition, we show that the expression of specific markers for each of these components is de-regulated in Pax-2 mutants. These data show that Pax-2 is required for multiple steps during the differentiation of intermediate mesoderm. In addition, Pax-2 mouse mutants provide an animal model for human hereditary kidney diseases.


Developmental trajectories of self-injurious behavior, suicidal behavior and substance misuse and their association with adolescent borderline personality pathology.
Adolescent risk-taking and self-harm behaviors are associated with affect dysregulation and impulsivity, both core features of borderline personality disorder (BPD). We hypothesized that the developmental courses of these behaviors i) tend to cluster rather than appear individually, and ii) might indicate adolescent BPD pathology. Therefore, we explored the developmental trajectories of self-injurious behavior (SIB), suicidal behavior (SB) and substance misuse (SM) in a community sample of adolescents; and we investigated the trajectories' overlap and its associations with BPD traits. 513 adolescents, aged 15-17 years, were followed for two years as part of the Saving and Empowering Young Lives in Europe study and its subsequent follow-up. Distinct developmental trajectories were explored using general growth mixture modeling. Three distinct classes were identified within each of the harmful behaviors SIB, SB and SM. Both the high-risk SIB trajectory and the high-risk SB trajectory demonstrated elevated initial degree of engagement, followed by a gradual decrease. The SM high-risk trajectory had a medium initial degree of engagement, which increased over time. There was a high degree of overlap (80-90%) among the high-risk trajectories for the three behaviors (SIB, SB and SM), and this overlap was significantly associated with elevated levels of BPD pathology. The data collection was based on participants' self-report. The findings indicate a similar pattern of reduction over time between SIB and SB for the high-risk trajectories, whereas the high-risk trajectories for SM show a pattern of increase over time. The observed symptom shift is associated with borderline personality pathology in adolescents. Therefore these behaviors might represent early indicators of risk supporting potential early detection.


Impact of resistance exercise training on interleukin-6 and JAK/STAT in young men.
The JAK/STAT signaling pathway is essential for myogenic regeneration and is regulated by a diverse range of ligands, including interleukin-6 (IL-6) and platelet-derived growth factor-BB (PDGF-BB). Our aim was to evaluate the responsiveness of IL-6 and PDGF-BB to intense exercise, along with STAT3 activation, before and after 12 weeks of resistance training. In young men, IL-6 and PDGF-BB protein concentrations were quantified in biopsied muscle and increased at 3 h post-exercise (17.5-fold and 3-fold, respectively). The response was unaltered by 12 weeks of training. Similarly, STAT3 phosphorylation was elevated post-exercise (12.5-fold), irrespective of training status, as was the expression of downstream targets c-MYC (8-fold), c-FOS (4.5-fold), and SOCS3 (2.3-fold). Thus, intense exercise transiently increases IL-6 and PDGF-BB proteins, and STAT3 phosphorylation is increased. These responses are preserved after intense exercise. This suggests they are not modified by training and may be an essential component of the adaptive responses to intense exercise.


Language extinction and linguistic fronts.
Language diversity has become greatly endangered in the past centuries owing to processes of language shift from indigenous languages to other languages that are seen as socially and economically more advantageous, resulting in the death or doom of minority languages. In this paper, we define a new language competition model that can describe the historical decline of minority languages in competition with more advantageous languages. We then implement this non-spatial model as an interaction term in a reaction-diffusion system to model the evolution of the two competing languages. We use the results to estimate the speed at which the more advantageous language spreads geographically, resulting in the shrinkage of the area of dominance of the minority language. We compare the results from our model with the observed retreat in the area of influence of the Welsh language in the UK, obtaining a good agreement between the model and the observed data.


Juridification of maternal deaths in Ethiopia: a study of the Maternal and Perinatal Death Surveillance and Response (MPDSR) system.
Juridification of maternal health care is on the rise globally, but little is known about its manifestations in resource constrained settings in sub-Saharan Africa. The Maternal and Perinatal Death Surveillance and Response (MPDSR) system is implemented in Ethiopia to record and review all maternal and perinatal deaths, but underreporting of deaths remains a major implementation challenge. Fear of blame and malpractice litigation among health workers are important factors in underreporting, suggestive of an increased juridification of birth care. By taking MPDSR implementation as an entry point, this article aims to explore the manifestations of juridification of birth care in Ethiopia. Based on multi-sited fieldwork involving interviews, document analysis and observations at different levels of the Ethiopian health system, we explore responses to maternal deaths at various levels of the health system. We found an increasing public notion of maternal deaths being caused by malpractice, and a tendency to perceive the juridical system as the only channel to claim accountability for maternal deaths. Conflicts over legal responsibility for deaths influenced birth care provision. Both health workers and health bureaucrats strived to balance conflicting concerns related to the MPDSR system: reporting all deaths vs revealing failures in service provision. This dilemma encouraged the development of strategies to avoid personalized accountability for deaths. In this context, increased juridification impacted both care and reporting practices. Our study demonstrates the need to create a system that secures legal protection of health professionals reporting maternal deaths as prescribed and provides the public with mechanisms to claim accountability and high-quality birth care services.


Influence of psychosocial, psychogenic and neurogenic stressors on several aspects of immune functioning in mice.
Analysis of stressor effects on immune functioning is complicated by the fact that the nature of the changes observed may be influenced by organismic factors (e.g., species, strain, age), the nature, severity and chronicity of the stressor, as well as the specific immune parameters being examined. It is demonstrated in the present investigation that in the highly reactive inbred BALB/cByJ mouse, the relatively hardy C57BL/6ByJ strain, as well as in the noninbred CD-1 strain, acute psychogenic (predator exposure) and neurogenic (footshock) stressors reduced splenic macrophage activity, and this effect was less marked after a chronic stressor. With protracted, but not transient, psychosocial disturbances (isolated housing) similar effects were seen, suggesting that the effect was not simply due to a change of the social mileau. The psychogenic and neurogenic stressors also enhanced LPS-stimulated lymphocyte proliferation in CD-1, but not in the inbred strains. However, isolated housing reduced both B and T cell proliferation, indicating that social isolation likely affects processes distinct from those of other stressors. Interestingly, when the aversiveness of the psychogenic stressor was increased (by decreasing the distance between the rat and the mouse) LPS-stimulated lymphocyte proliferation was reduced in the reactive BALB/cByJ strain, but increased in the hardy C57BL/6ByJ mice. This stressor, however, enhanced T cell proliferation in both strains of mice. It is suggested that analysis of stressor effects need to consider in greater detail the characteristics of the organism being stressed, as well as the characteristics of the stressor itself.


Asymmetric effects of emotion on mnemonic interference.
Emotional experiences can strengthen memories so that they can be used to guide future behavior. Emotional arousal, mediated by the amygdala, is thought to modulate storage by the hippocampus, which may encode unique episodic memories via pattern separation--the process by which similar memories are stored using non-overlapping representations. While prior work has examined mnemonic interference due to similarity and emotional modulation of memory independently, examining the mechanisms by which emotion influences mnemonic interference has not been previously accomplished in humans. To this end, we developed an emotional memory task where emotional content and stimulus similarity were varied to examine the effect of emotion on fine mnemonic discrimination (a putative behavioral correlate of hippocampal pattern separation). When tested immediately after encoding, discrimination was reduced for similar emotional items compared to similar neutral items, consistent with a reduced bias towards pattern separation. After 24h, recognition of emotional target items was preserved compared to neutral items, whereas similar emotional item discrimination was further diminished. This suggests a potential mechanism for the emotional modulation of memory with a selective remembering of gist, as well as a selective forgetting of detail, indicating an emotion-induced reduction in pattern separation. This can potentially increase the effective signal-to-noise ratio in any given situation to promote survival. Furthermore, we found that individuals with depressive symptoms hyper-discriminate negative items, which correlated with their symptom severity. This suggests that utilizing mnemonic discrimination paradigms allows us to tease apart the nuances of disorders with aberrant emotional mnemonic processing.


Attitudes of people in the UK with HIV who Are Antiretroviral (ART) Naïve to starting ART at high CD4 counts for potential health benefit or to prevent HIV transmission.
To assess if a strategy of early ART to prevent HIV transmission is acceptable to ART naïve people with HIV with high CD4 counts. ASTRA is a UK multicentre, cross sectional study of 3258 HIV outpatients in 2011/12. A self-completed questionnaire collected sociodemographic, behavioral and health data, and attitudes to ART; CD4 count was recorded from clinical records. ART naïve participants with CD4 ≥350 cells/µL (n = 281) were asked to agree/disagree/undecided with the statements (i) I would want to start treatment now if this would slightly reduce my risk of getting a serious illness, and (ii) I would want to start treatment now if this would make me less infectious to a sexual partner, even if there was no benefit to my own health. Participants were 85% MSM, 76% white, 11% women. Of 281 participants, 49.5% and 45.2% agreed they would start ART for reasons (i) and (ii) respectively; 62.6% agreed with either (i) or (ii); 12.5% agreed with neither; 24.9% were uncertain. Factors independently associated (p<0.1) with agreement to (i) were: lower CD4, more recent HIV diagnosis, physical symptoms, not being depressed, greater financial hardship, and with agreement to (ii) were: being heterosexual, more recent HIV diagnosis, being sexually active. A strategy of starting ART at high CD4 counts is likely to be acceptable to the majority of HIV-diagnosed individuals. Almost half with CD4 >350 would start ART to reduce infectiousness, even if treatment did not benefit their own health. However a significant minority would not like to start ART either for modest health benefit or to reduce infectivity. Any change in approach to ART initiation must take account of individual preferences. Transmission models of potential benefit of early ART should consider that ART uptake may be lower than that seen with low CD4 counts.


COVID-19 risk perception and coping mechanisms: Does gender make a difference?
The novel coronavirus disease (COVID-19) emerged as a real threat to humans, drastically disrupting everyday life in 2020-21. Although the pandemic affected people from all walks of life, irrespective of age or gender, the way the risk is perceived varies from one person to another. The pandemic risk reduction strategies can only be effective if individuals and communities respond positively to them, and for that, it is important to understand how the risk is perceived and responded to, differently by different groups of people. Gender plays a vital role in shaping risk perceptions and coping strategies, reflecting the predisposition of the public to accept health interventions and take precautionary measures. This study aims to understand the gender differences in COVID-19 risk perception and coping mechanisms - Pakistan is selected as a case study area. Following on from designing the questionnaire, which included 40 indicators grouped into domains (four risk perception and three coping mechanisms domains), an online survey was conducted, and a sample of 389 respondents was collected (248 male and 141 female). An index-based approach was used to quantify risk perception and its domains (fear, behaviour, awareness, and trust), and likewise coping mechanisms and their domains (problem, emotion, and action). Statistical tests were employed to ascertain the differences among both genders, whereas regression modelling was used to measure the effect of gender on overall risk perception and coping mechanisms. Results reveal that perceived fear and trust varied significantly between Pakistani men and women, while coping mechanisms were also notably different between the two genders. Females were found to perceive risks higher, complied more with the government's guidelines, and coped better than males in response to COVID-19. These findings imply that the gender aspect must be incorporated in designing effective communication and risk reduction strategies to efficiently address the current and potential future pandemic situations.


Molecular evolution of glutamine synthetase II: Phylogenetic evidence of a non-endosymbiotic gene transfer event early in plant evolution.
Glutamine synthetase (GS) is essential for ammonium assimilation and the biosynthesis of glutamine. The three GS gene families (GSI, GSII, and GSIII) are represented in both prokaryotic and eukaryotic organisms. In this study, we examined the evolutionary relationship of GSII from eubacterial and eukaryotic lineages and present robust phylogenetic evidence that GSII was transferred from gamma-Proteobacteria (Eubacteria) to the Chloroplastida. GSII sequences were isolated from four species of green algae (Trebouxiophyceae), and additional green algal (Chlorophyceae and Prasinophytae) and streptophyte (Charales, Desmidiales, Bryophyta, Marchantiophyta, Lycopodiophyta and Tracheophyta) sequences were obtained from public databases. In Bayesian and maximum likelihood analyses, eubacterial (GSIIB) and eukaryotic (GSIIE) GSII sequences formed distinct clades. Both GSIIB and GSIIE were found in chlorophytes and early-diverging streptophytes. The GSIIB enzymes from these groups formed a well-supported sister clade with the gamma-Proteobacteria, providing evidence that GSIIB in the Chloroplastida arose by horizontal gene transfer (HGT). Bayesian relaxed molecular clock analyses suggest that GSIIB and GSIIE coexisted for an extended period of time but it is unclear whether the proposed HGT happened prior to or after the divergence of the primary endosymbiotic lineages (the Archaeplastida). However, GSIIB genes have not been identified in glaucophytes or red algae, favoring the hypothesis that GSIIB was gained after the divergence of the primary endosymbiotic lineages. Duplicate copies of the GSIIB gene were present in Chlamydomonas reinhardtii, Volvox carteri f. nagariensis, and Physcomitrella patens. Both GSIIB proteins in C. reinhardtii and V. carteri f. nagariensis had N-terminal transit sequences, indicating they are targeted to the chloroplast or mitochondrion. In contrast, GSIIB proteins of P. patens lacked transit sequences, suggesting a cytosolic function. GSIIB sequences were absent in vascular plants where the duplication of GSIIE replaced the function of GSIIB. Phylogenetic evidence suggests GSIIB in Chloroplastida evolved by HGT, possibly after the divergence of the primary endosymbiotic lineages. Thus while multiple GS isoenzymes are common among members of the Chloroplastida, the isoenzymes may have evolved via different evolutionary processes. The acquisition of essential enzymes by HGT may provide rapid changes in biochemical capacity and therefore be favored by natural selection.


Use and mechanism of action of AS101 in protecting bone marrow colony forming units-granulocyte-macrophage following purging with ASTA-Z 7557.
Ammonium trichloro(dioxoethylene-O,O')tellurate (AS101) has been shown previously to provide radioprotective effects when given to mice 24 h prior to irradiation and to protect mice from lethal and sublethal doses of cyclophosphamide (CTX). In this study we examined the ability of AS101 to protect mice bone marrow colony forming units-granulocyte-macrophage treated in vitro with various doses of ASTA-Z 7557, a potent derivative of cyclophosphamide. We demonstrate that prior incubation with AS101 protects colony forming units-granulocyte-macrophage from toxic effects of ASTA-Z. This protection can also be conferred by injection of mice with AS101 prior to incubation of their bone marrow in vitro with ASTA-Z. Prior incubation with AS101 was shown not to protect K562 leukemic cells or HL-60 cells from the toxic effects of ASTA-Z. We show that AS101 protection from the toxic effects of ASTA-Z in vitro and CTX in vivo can be partially ascribed to increased aldehyde dehydrogenase (ALDH) activity induced by AS101. This was shown directly by measuring cellular ALDH activity and indirectly by measuring the toxicity of ASTA-Z and CTX in the presence of cyanamide, an inhibitor of ALDH. AS101 is also demonstrated in this study to protect spleen cells from the toxic effects of 5-fluorouracil, probably through a different mechanism. These properties of AS101 make it a useful candidate for increasing the qualitative potential of bone marrow used for autologous transplantation after purging with ASTA-Z. In addition, the results suggest an increase in ALDH activity by AS101 as one of the mechanisms of protection from the toxic effects of ASTA-Z and CTX. However, the chemoprotectiveness of AS101 was found not to be restricted to cyclophosphamide, since as shown in this study, AS101 helped by other mechanisms to reconstitute the number of spleen cells after 5-fluorouracil treatment.


Impact of palliative home care support on the quality and costs of care at the end of life: a population-level matched cohort study.
To evaluate the impact of palliative home care support on the quality of care and costs in the last 14 days of life. Matched cohort study using linked administrative databases. All people who died in Belgium in 2012 (n=107 847). 8837 people who received palliative home care support in the last 720 to 15 days of life matched 1:1 by propensity score to 8837 people who received usual care. Receiving the allowance for palliative home patients, multidisciplinary palliative home care team visit or palliative nurse or physiotherapist visit at home. Home death, number of family physician contacts, number of primary caregiver contacts, hospital death, hospital admission, intensive care unit (ICU) admission, emergency department (ED) admission, diagnostic testing, blood transfusion and surgery. Total inpatient and outpatient costs. All outcomes were measured in the last 14 days of life. In the unmatched cohort, 11 149 (13.5%) people received palliative home care support in the last 720 to 15 days of life. After matching, those using palliative home care support had, compared with those who did not, more family physician contacts (mean 3.1 [SD=6.5] vs 0.8 [SD=1.2]), more chance of home death (56.2%vs13.8%; relative risk [RR]=4.08, 95% CI 3.86 to 4.31), lower risk of hospital admission (27.4%vs60.8%; RR=0.45, 95% CI 0.43 to 0.46), ICU admission (18.3%vs40.4%; RR=0.45, 95% CI 0.43 to 0.48) or ED admission (15.2%vs28.1%; RR=0.54, 95% CI 0.51 to 0.57). Mean total costs of care were lower for those using palliative home care support (€3081 [95% CI €3025 to €3136] vs €4698 [95% CI €4610 to €4787]; incremental cost: -€1617 [p<0.001]). Palliative home care support use positively impacts quality of care and reduces total costs of care at the end of life in Belgium. Policy makers and healthcare practitioners should increasingly focus on communicating the existing options for palliative home care support to patients and their caregivers.


Body mass index and use and costs of primary care services among women aged 55-79 years in England: a cohort and linked data study.
Excess weight is associated with poor health and increased healthcare costs. There are no reliable data describing the association between BMI and the use and costs of primary care services in the United Kingdom. Among 69,440 participants in the Million Women Study with primary care records in the Clinical Practice Research Datalink between April 2006 (mean age 64 years) and March 2014, the annual rates and costs of their primary care consultations, prescription medications, and diagnostic and monitoring tests were estimated in relation to their self-reported body mass index (BMI) at recruitment in 1996-2001 (mean age 56 years). Associations of BMI with annual costs were projected to all women in England aged 55-79 years in 2013. Over an average follow-up of 6.0 years, annual rates and mean costs were lowest for women with a BMI of 20 to <22.5 kg/m2 for consultations (7.0 consultations, 99% CI 6.8-7.1; £288, £280-£295) and prescription medications (27.0 prescribed items, 26.0-27.9; £227, £216-£237). Above 20 kg/m2, a 2 kg/m2 higher BMI (a 5 kg change in weight for a woman of average height) was associated with 5.2% (4.8-5.6) and 9.9% (9.2-10.6) higher mean annual consultation and prescription medication costs, respectively. Annual rates and mean costs of diagnostic and monitoring tests were similar for women with different BMIs. Among all women aged 55-79 years in England, excess weight accounted for an estimated 11% (£229 million/£2.2 billion) of all consultation costs and 20% (£384 million/£1.9 billion) of all prescription medication costs, of which 27% were for diabetes drugs, 19% for circulatory system drugs, and 13% for analgesics. Excess body weight is associated with higher use and costs of primary care services among women in England. Reducing the prevalence of excess weight could improve the health of women and reduce pressures on primary care.


Environmental Tobacco Smoke Exposure and Survival Following Breast Cancer.
Environmental tobacco smoke (ETS) exposure is hypothesized to influence survival after breast cancer, but few studies have examined this association. A population-based cohort of women (N = 1,508) diagnosed with first primary invasive or in situ breast cancer in 1996 to 1997 was interviewed shortly after diagnosis and again approximately 5 years later to assess ETS exposure, and women were followed for more than 18 years using the National Death Index; 597 deaths (237 associated with breast cancer) were identified. Multivariable Cox regression was used to estimate adjusted HRs and 95% confidence intervals (CI) for mortality among women with breast cancer as related to at-diagnosis and at-/postdiagnosis changes in ETS exposure. There was little or no association between at-diagnosis ETS exposure and all-cause (HR = 1.04; 95% CI, 0.78-1.40) or breast cancer-specific (HR = 0.98; 95% CI, 0.63-1.52) mortality. Mortality was elevated among women who reported cessation in postdiagnosis ETS exposure up to 1 year before the follow-up assessment, for all-cause (HR = 1.81; 95% CI, 0.87-3.74) and breast cancer mortality (HR = 1.89; 95% CI, 0.68-5.24); however, estimates were imprecise. We found little evidence of an association between at-diagnosis ETS exposure and mortality after breast cancer. Postdiagnosis cessation of ETS exposure was positively associated with mortality, although we could not rule out chance and reverse causation as possible explanations. Exposure to ETS does not appear to influence mortality after breast cancer. Cancer Epidemiol Biomarkers Prev; 26(2); 278-80. ©2016 AACR.


Comprehensive in vivo mapping of the human basal ganglia and thalamic connectome in individuals using 7T MRI.
Basal ganglia circuits are affected in neurological disorders such as Parkinson's disease (PD), essential tremor, dystonia and Tourette syndrome. Understanding the structural and functional connectivity of these circuits is critical for elucidating the mechanisms of the movement and neuropsychiatric disorders, and is vital for developing new therapeutic strategies such as deep brain stimulation (DBS). Knowledge about the connectivity of the human basal ganglia and thalamus has rapidly evolved over recent years through non-invasive imaging techniques, but has remained incomplete because of insufficient resolution and sensitivity of these techniques. Here, we present an imaging and computational protocol designed to generate a comprehensive in vivo and subject-specific, three-dimensional model of the structure and connections of the human basal ganglia. High-resolution structural and functional magnetic resonance images were acquired with a 7-Tesla magnet. Capitalizing on the enhanced signal-to-noise ratio (SNR) and enriched contrast obtained at high-field MRI, detailed structural and connectivity representations of the human basal ganglia and thalamus were achieved. This unique combination of multiple imaging modalities enabled the in-vivo visualization of the individual human basal ganglia and thalamic nuclei, the reconstruction of seven white-matter pathways and their connectivity probability that, to date, have only been reported in animal studies, histologically, or group-averaged MRI population studies. Also described are subject-specific parcellations of the basal ganglia and thalamus into sub-territories based on their distinct connectivity patterns. These anatomical connectivity findings are supported by functional connectivity data derived from resting-state functional MRI (R-fMRI). This work demonstrates new capabilities for studying basal ganglia circuitry, and opens new avenues of investigation into the movement and neuropsychiatric disorders, in individual human subjects.


Structure of the APPL1 BAR-PH domain and characterization of its interaction with Rab5.
APPL1 is an effector of the small GTPase Rab5. Together, they mediate a signal transduction pathway initiated by ligand binding to cell surface receptors. Interaction with Rab5 is confined to the amino (N)-terminal region of APPL1. We report the crystal structures of human APPL1 N-terminal BAR-PH domain motif. The BAR and PH domains, together with a novel linker helix, form an integrated, crescent-shaped, symmetrical dimer. This BAR-PH interaction is likely conserved in the class of BAR-PH containing proteins. Biochemical analyses indicate two independent Rab-binding sites located at the opposite ends of the dimer, where the PH domain directly interacts with Rab5 and Rab21. Besides structurally supporting the PH domain, the BAR domain also contributes to Rab binding through a small surface region in the vicinity of the PH domain. In stark contrast to the helix-dominated, Rab-binding domains previously reported, APPL1 PH domain employs beta-strands to interact with Rab5. On the Rab5 side, both switch regions are involved in the interaction. Thus we identified a new binding mode between PH domains and small GTPases.


Uniform field loop-gap resonator and rectangular TEU02 for aqueous sample EPR at 94GHz.
In this work we present the design and implementation of two uniform-field resonators: a seven-loop-six-gap loop-gap resonator (LGR) and a rectangular TEU02 cavity resonator. Each resonator has uniform-field-producing end-sections. These resonators have been designed for electron paramagnetic resonance (EPR) of aqueous samples at 94GHz. The LGR geometry employs low-loss Rexolite end-sections to improve the field homogeneity over a 3mm sample region-of-interest from near-cosine distribution to 90% uniform. The LGR was designed to accommodate large degassable Polytetrafluorethylen (PTFE) tubes (0.81mm O.D.; 0.25mm I.D.) for aqueous samples. Additionally, field modulation slots are designed for uniform 100kHz field modulation incident at the sample. Experiments using a point sample of lithium phthalocyanine (LiPC) were performed to measure both the uniformity of the microwave magnetic field and 100kHz field modulation, and confirm simulations. The rectangular TEU02 cavity resonator employs over-sized end-sections with sample shielding to provide an 87% uniform field for a 0.1×2×6mm3 sample geometry. An evanescent slotted window was designed for light access to irradiate 90% of the sample volume. A novel dual-slot iris was used to minimize microwave magnetic field perturbations and maintain cross-sectional uniformity. Practical EPR experiments using the application of light irradiated rose bengal (4,5,6,7-tetrachloro-2',4',5',7'-tetraiodofluorescein) were performed in the TEU02 cavity. The implementation of these geometries providing a practical designs for uniform field resonators that continue resonator advancements towards quantitative EPR spectroscopy.


De novo characterization of Japanese scallop Mizuhopecten yessoensis transcriptome and analysis of its gene expression following cadmium exposure.
Japanese scallop has been cultured on a large-scale in China for many years. However, serious marine pollution in recent years has resulted in considerable loss to this industry. Moreover, due to the lack of genomic resources, limited research has been carried out on this species. To facilitate the understanding at molecular level immune and stress response mechanism, an extensive transcriptomic profiling and digital gene expression (DGE) database of Japanese scallop upon cadmium exposure was carried out using the Illumina sequencing platform. RNA-seq produced about 112 million sequencing reads from the tissues of adult Japanese scallops. These reads were assembled into 194,839 non-redundant sequences with open reading frame (ORF), of which 14,240 putative amino acid sequences were assigned biological function annotation and were annotated with gene ontology and eukaryotic orthologous group terms. In addition, we identified 720 genes involved in response to stimulus and 302 genes involved in immune-response pathways. Furthermore, we investigated the transcriptomic changes in the gill and digestive gland of Japanese scallops following cadmium exposure using a tag-based DGE system. A total of 7,556 and 3,002 differentially expressed genes were detected, respectively, and functionally annotated with KEGG pathway annotations. This study provides a comprehensive transcripts sequence resource for the Japanese scallop and presents a survey of gene expression in response to heavy metal exposure in a non-model marine invertebrate via the Illumina sequencing platform. These results may contribute to the in-depth elucidation of the molecular mechanisms involved in bivalve responses to marine pollutants.


Antitoxin activity of aqueous extract of Cyclea peltata root against Naja naja venom.
Snakebites are a significant and severe global health problem. Till date, anti-snake venom serum is the only beneficial remedy existing on treating the snakebite victims. As antivenom was reported to induce early or late adverse reactions to human beings, snake venom neutralizing potential for Cyclea peltata root extract was tested for the present research by ex vivo and in vivo approaches on Naja naja toxin. Ex vivo evaluation of venom toxicity and neutralization assays was carried out. The root extracts from C. peltata were used to evaluate the Ex vivo neutralization tests such as acetylcholinesterase, protease, direct hemolysis assay, phospholipase activity, and procoagulant activity. Gas chromatography-mass spectrometry (GC-MS) analysis from root extracts of C. peltata was done to investigate the bioactive compounds. The in vivo calculation of venom toxicity (LD50) of N. naja venom remained to be 0.301 μg. C. peltata root extracts were efficiently deactivated the venom lethality, and effective dose (ED50) remained to be 7.24 mg/3LD50 of N. naja venom. C. peltata root extract was found effective in counteracting all the lethal effects of venom. GC-MS analysis of the plant extract revealed the presence of antivenom compounds such as tetradecanoic and octadecadienoic acid which have neutralizing properties on N. naja venom. The result from the ex vivo and in vivo analysis indicates that C. peltata plant root extract possesses significant compounds such as tetradecanoic acid hexadecanoic acid, heptadecanoic acid, and octadecadienoic acid which can counteract the toxins present in N. naja.


Behavioral changes after long-term exposure to organic solvents and their mixtures: determining factors and research results.
This presentation summarizes the characteristics determining the psychological end state after long-term solvent exposure. It also presents the results of research on changes in psychological functions after long-term solvent exposure. Such factors as differential vulnerability and environmental influences other than solvent exposure at work and elsewhere affect the end state. The results of five different Finnish studies are summarized. These studies concerned workers with solvent poisoning, car painters exposed to solvent mixtures, rayon viscose workers exposed to carbon disulfide, printers exposed to toluene, and laminators exposed to styrene. Intellectual functions, short-term memory, and sensory and motor functions were inspected. Verbal intelligence was lowered only in cases of intoxication and exposure to mixtures of organic solvents. Visuoconstructive abilities were deteriorated in those groups with verbal intelligence deterioration, and also among both rayon viscose workers with solvent poisoning and car painters. The group exposed to styrene showed functional changes only in visuomotor accuracy. Lowered manual or finger dexterity was found among workers exposed to carbon disulfide, workers with solvent poisoning, and car painters. The most disturbances were accumulated among those exposed to mixtures of organic solvents.


Structural characterisation of the capsular polysaccharide expressed by Burkholderia thailandensis strain E555:: wbiI (pKnock-KmR) and assessment of the significance of the 2-O-acetyl group in immune protection.
Burkholderia pseudomallei and its close relative B. mallei are human pathogens that are classified as Tier 1 bio-threat agents. Both organisms have previously been shown to constitutively produce a capsular polysaccharide (CPS) that is both a virulence determinant and protective antigen. Extraction and purification of CPS for use as a potential vaccine candidate requires containment level 3 laboratories which is expensive and time-consuming. B. thailandensis strain E555 is closely related to B. pseudomallei and B. mallei, but is non-pathogenic to humans and based on immunological cross-reactivity has previously been shown to express a B. pseudomallei-like CPS. In this study, capsular polysaccharide isolated from an O-antigen deficient strain of B. thailandensis E555 was identified by 1H and 13C NMR spectroscopy as -3-)-2-O-acetyl-6-deoxy-β-d-manno-heptopyranose-(-1, and identical to that produced by B. pseudomallei. This was further substantiated by anti-CPS monoclonal antibody binding. In connection with the production of CPS fragments for use in glycoconjugate vaccines, we set out to assess the importance or otherwise of the CPS 2-OAc groups in immune protection. To this end conjugates of the native and de-O-acetylated CPS with the Hc fragment of tetanus toxin (TetHc) were used as vaccines in a mouse model of melioidosis. The level of protection provided by deacetylated CPS was significantly lower than that from native, acetylated CPS. In addition, sera from mice vaccinated with the deacetylated CPS conjugate did not recognise native CPS. This suggests that CPS extracted from B. thailandensis can be used as antigen and that the acetyl group is essential for protection.


Five-year changes in North Carolina outpatient cardiac rehabilitation.
The purpose of this study was to describe cardiac rehabilitation programming, barriers to participation, and reasons for dropout in North Carolina from a program director's perspective and to compare those results with those of a similar statewide survey conducted 5 years earlier. In 1999 and 2004, a survey was mailed to all North Carolina program directors of outpatient cardiac rehabilitation programs. The response rate was 85% (61/72) in 1999 and 79% (61/77) in 2004. More than 85% of North Carolinians older than 40 years lived within a 15-mile buffer of an outpatient cardiac rehabilitation program in 2004. Most programs were staffed with personnel trained in nursing, exercise physiology, and nutrition in 1999 and 2004. Women and African Americans remained disproportionately underrepresented as participants in the program for both years. In 2004, approximately one third of cardiac rehabilitation programs reported having a referral to rehabilitation on the hospital discharge plan for myocardial infarction and coronary artery bypass surgery. In 1999 and 2004, the most frequently reported barrier to participation remained financial, followed by lack of interest or motivation and workplace conflicts. Work conflicts, lack of interest, and comorbidities were the most frequently reported reasons for dropping out from cardiac rehabilitation programs in both 1999 and 2004. Increasing participation in cardiac rehabilitation programs by addressing barriers at multiple levels may facilitate greater patient participation. This statewide survey could be used in other states as a surveillance tool, to track changes in rehabilitation over time from a program director's perspective.


Prophylactic and therapeutic efficacy of nitazoxanide against Cryptosporidium parvum in experimentally challenged neonatal calves.
Diarrhoea caused by Cryptosporidium parvum is a major problem in calves younger than 4 weeks of age. To date only a few compounds have been approved for prophylactic and none for therapeutic use. Nitazoxanide (NTZ) has proven its efficacy in vitro against C. parvum and is approved by FDA for the treatment of human cryptosporidiosis. In a first experimental study, 3 uninfected calves were treated with NTZ and pharmacokinetics was followed through blood samples. Serum samples of uninfected treated calves contained both NTZ metabolites (tizoxanide and tizoxanide glucuronide) and oral administration at 12 h intervals was considered as optimal. Three groups of three calves (1-3 days old) were then each inoculated with 1x10(7) oocysts of C. parvum (cattle genotype): the prophylactic group received 15 mg/kg body weight NTZ twice daily orally in milk from 1 day before to 8 days postinoculation (dpi). The therapeutic group received the same dosage of NTZ for 10 days from the appearance of diarrhoea (between 1 and 5 dpi). The control group was left untreated. All calves were monitored daily from day -1 to 28 dpi and faecal samples were collected for evaluation of consistency and for determination of oocyst numbers per gram (OPG) of faeces. Diarrhoea was observed in all calves within the first week. Neither prophylactic nor therapeutic use of NTZ improved the clinical appearance and calves of the therapeutic showed a longer diarrheic episode (p<0.05) with strong altered faecal consistency compared to the untreated control group. The number of days with oocyst excretion did not differ significantly between the groups. In 5 out of 6 infected and treated calves oocyst excretion stopped only after discontinuation of treatment. In the prophylactic and in the control group mean values of the sum of the daily OPG per calf (8.5x10(6) and 8.0x10(6), respectively) and of the mean daily number of OPG (0.3x10(6) and 0.3x10(6), respectively) were similar, while the therapeutic group showed significantly lower values (1.9x10(6) and 0.06x10(6), respectively, p<0.05). However oocyst determinations in this group may have been altered by the severe diarrhoea, diluting oocyst densities in the analysed faecal samples. In conclusion, these preliminary results about the first prophylactic and therapeutic use of NTZ in calves did not show the expected positive effect on the course of the Cryptosporidium-infection, neither on reducing the clinical severity, nor on oocyst excretion.


Interleukin-2 transcriptional block by multifunctional Ca2+/calmodulin kinase.
In the presence of costimulation, Ca2+ influx in T cells leads to activation (transcription of interleukin-2; ref. 2) via calcineurin. In the absence of costimulation, Ca2+ influx results in anergy (interleukin-2 transcriptional block) through an unknown mechanism. Specific attenuation of interleukin-2 transcriptional induction occurs in Jurkat T cells following pretreatment with a Ca2+ ionophore. A > 90% block of inducible interleukin-2 reporter gene activity was initiated by transfection of a constitutively active mutant of multifunctional Ca2+/calmodulin-dependent protein kinase (CaM kinase or CaM kinase II), but not by constitutive mutants of CaM kinase IV, calcineurin or protein kinase C. The block was complete six hours after kinase transfection and showed specificity for interleukin-2; there was no change in beta-actin transcription or in c-fos transcription induced by phorbol myristyl acetate, and a Rous sarcoma virus promoter was stimulated threefold. Multifunctional CaM kinase also attenuated interleukin-2 activation by calcineurin plus phorbol ester. T-cell receptor signalling activates multifunctional CaM kinase. These findings suggest that two Ca2+/calmodulin-responsive enzymes, multifunctional CaM kinase and calcineurin, could mediate the divergent effects of Ca2+ signals in T-lymphocyte regulation.


Diabetes-related major lower limb amputation incidence is strongly related to diabetic foot service provision and improves with enhancement of services: peer review of the South-West of England.
To investigate the relationship between high diabetes-related lower limb amputation incidence and foot care services in the South-West region of England. The introduction of 10 key elements of foot care service provision in one area of the South-West resulted in stabilization of foot ulcer incidence and sustained reduction in amputation incidence from 2007. Services introduced included administrative support, standardized general practice foot screening, improved community podiatry staffing, hospital multidisciplinary foot clinics, effective care pathways, availability of an orthotist and audit. Peer reviews of the region's diabetes foot care services were undertaken to assess delivery of these service provisions and compare this with major amputation incidence in other regions with data provided by Yorkshire and Humber Public Health Observatory Hospital Episode Statistics. Recommendations were made to improve service provision. In 2015 changes in service provision and amputation incidence were reviewed. Initial reviews in 2013 showed that the 3-year diabetes-related major amputation incidence correlated inversely with adequate delivery of diabetes foot care services (P=0.0024, adjusted R2 =0.51). Repeat reviews in 2015 found that two or more foot care service improvements were reported by six diabetes foot care providers, with improvement in outcomes. The negative relationship between major amputation incidence and service provision remained strong both in the period 2012-2015 and in the year 2015 only (P ≤0.0012, adjusted R2 =0.56, and P= 0.0005, R2 =0.62, respectively). Major diabetes-related lower limb amputation incidence is significantly inversely correlated with foot care services provision. Introduction of more effective service provision resulted in significant reductions in major amputation incidence within 2 years. Failure to improve unsatisfactory service provision resulted in continued high amputation incidence.


Serum microRNA-21 and microRNA-221 as potential biomarkers for cerebrovascular disease.
MicroRNA miR-21, miR-221 and miR-145 have been implicated in the cardiovascular system. We aimed to compare the serum levels of the three microRNAs (miRNAs) in different severities of cerebrovascular diseases and evaluate the feasibility of using these miRNAs as biomarkers for stroke. We enrolled 167 subjects with ischemic stroke, 66 atherosclerosis subjects with any carotid plaque score and 157 healthy controls. These three types of subjects represent three levels of severity in cerebrovascular diseases. Analysis of covariance was used to evaluate the relationship between miRNAs and disease severity with adjustment for conventional risk factors. To test the prediction for stroke, we built regression models containing the serum miRNA levels and risk factors. Prediction capabilities were compared by the receiver operating characteristic curves. Stroke patients and atherosclerosis subjects had significantly higher miR-21 and lower miR-221 serum levels than healthy controls, while the miR-145 expression was too low to provide useful information in this regard. The best model showed that miR-21 and miR-221 were independent predictors. There was a 6.2-fold increase for stroke risk when miR-21 levels increase by log₁₀2(-ΔCt) = 1, while a 10.4-fold increase was observed as miR-221 decreases by log₁₀2(-ΔCt) = 1. Serum miR-145 was not detected in over 50% of the patients and it may not be an ideal marker to predict stroke. MiR-21 and miR-221 are novel biomarkers for atherosclerosis and stroke.


Efficacy of silymarin-phospholipid complex in reducing the toxicity of aflatoxin B1 in broiler chicks.
Silymarin, the standardized extract of Silybum marianum, is used as a hepatoprotector in man, and is a potent antihepatotoxic agent. This study focused on the effects of a silymarin-phospholipid complex in reducing the toxic effects of aflatoxin B1 (AFB1) in broiler chickens. Twenty-one 14-d-old male commercial broilers were randomly allotted to 3 groups and treated as follows: basal diet alone [Group C (Control)]; AFB1 at 0.8 mg/kg of feed [Group B1]; AFB1 at 0.8 mg/kg of feed plus silymarin phytosome, a silymarin complexed form with phospholipids from soy, at 600 mg/kg of BW [Group B1+Sil]. Considering the whole growth cycle, BW gain and feed intake were lower in AFB1-treated birds with respect to controls (P < 0.05). In the B1+Sil group, BW gain and feed intake were higher with respect to birds receiving AFB1 alone (P < 0.05), and not different from the control birds. Serum biochemistry showed no difference among groups, except for a decrease of alanine amino transferase (ALT) in chicks treated only with AFB1. Alanine amino transferase activity in AFB1 plus silymarin phytosome treated birds was not different from the controls. No treatment differences were noted on liver weight. In conclusion, our results suggest that silymarin phytosome can provide protection against the negative effects of AFB1 on performance of broiler chicks.


Variations in apolipoprotein E frequency with age in a pooled analysis of a large group of older people.
Variation in the apolipoprotein E gene (APOE) has been reported to be associated with longevity in humans. The authors assessed the allelic distribution of APOE isoforms ε2, ε3, and ε4 among 10,623 participants from 15 case-control and cohort studies of age-related macular degeneration (AMD) in populations of European ancestry (study dates ranged from 1990 to 2009). The authors included only the 10,623 control subjects from these studies who were classified as having no evidence of AMD, since variation within the APOE gene has previously been associated with AMD. In an analysis stratified by study center, gender, and smoking status, there was a decreasing frequency of the APOE ε4 isoform with increasing age (χ(2) for trend = 14.9 (1 df); P = 0.0001), with a concomitant increase in the ε3 isoform (χ(2) for trend = 11.3 (1 df); P = 0.001). The association with age was strongest in ε4 homozygotes; the frequency of ε4 homozygosity decreased from 2.7% for participants aged 60 years or less to 0.8% for those over age 85 years, while the proportion of participants with the ε3/ε4 genotype decreased from 26.8% to 17.5% across the same age range. Gender had no significant effect on the isoform frequencies. This study provides strong support for an association of the APOE gene with human longevity.


Inverse solvent effects in the heterogeneous and homogeneous epoxidation of cis-2-heptene with [2-percarboxyethyl]-functionalized silica and meta-chloroperbenzoic acid.
The rate constants for the epoxidation of cis-2-heptene with [2-percarboxyethyl]-functionalized silica (1a) and meta-chloroperbenzoic acid (mCPBA) (1b) in different solvents have been determined at temperatures in the -10 to 40 °C range. The heterogeneous epoxidation exhibits a dependence of the reaction rate on solvent polarity opposite to its homogeneous counterpart and anomalous activation parameters in n-hexane, which are interpreted in terms of the surface-promoted solvent structure at the solid-liquid interface. The results show that highly polar solvents can strongly inhibit heterogeneous reactions performed with silica-supported reagents or catalysts.


Extremely low frequency electromagnetic field and wound healing: implication of cytokines as biological mediators.
Wound healing is a highly coordinated and complex process involving various cell types, chemical mediators and the surrounding extracellular matrix, resulting in a tightly orchestrated re-establishment of tissue integrity by specific cytokines. It consists of various dynamic processes including a series of overlapping phases: inflammation, proliferation, re-epithelialization and remodeling. One of the underlying mechanisms responsible for the disturbances in wound healing is an out-of-control inflammatory response that can cause pathological consequences, such as hypertrophic scars, keloids or chronic wounds and ulcers. Recently, several reports have evaluated the effects of extremely low frequency electromagnetic fields (EMFs) on tissue repair. In particular, the data analysis supports an anti-inflammatory effect of EMFs by the modulation of cytokine profiles that drive the transition from a chronic pro-inflammatory state to an anti-inflammatory state of the healing process. In this review, we focus on the effect of EMFs on skin wound healing showing emerging details of the anti-inflammatory effects of EMFs, with a view to cytokines as candidate biomarkers. Molecular clarification of the mechanisms involved in the modulation of inflammatory factors following exposure to EMFs will provide a better understanding of the cellular responses induced by EMFs and a potential, additional treatment in non-responding, chronic wounds.


MesoTRAP: a feasibility study that includes a pilot clinical trial comparing video-assisted thoracoscopic partial pleurectomy decortication with indwelling pleural catheter in patients with trapped lung due to malignant pleural mesothelioma designed to address recruitment and randomisation uncertainties and sample size requirements for a phase III trial.
One of the most debilitating symptoms of malignant pleural mesothelioma (MPM) is dyspnoea caused by pleural effusion. MPM can be complicated by the presence of tumour on the visceral pleura preventing the lung from re-expanding, known as trapped lung (TL). There is currently no consensus on the best way to manage TL. One approach is insertion of an indwelling pleural catheter (IPC) under local anaesthesia. Another is video-assisted thoracoscopic partial pleurectomy/decortication (VAT-PD). Performed under general anaesthesia, VAT-PD permits surgical removal of the rind of tumour from the visceral pleura thereby allowing the lung to fully re-expand. MesoTRAP is a feasibility study that includes a pilot multicentre, randomised controlled clinical trial comparing VAT-PD with IPC in patients with TL and pleural effusion due to MPM. The primary objective is to measure the SD of visual analogue scale scores for dyspnoea following randomisation and examine the patterns of change over time in each treatment group. Secondary objectives include documenting survival and adverse events, estimating the incidence and prevalence of TL in patients with MPM, examining completion of alternative forms of data capture for economic evaluation and determining the ability to randomise 38 patients in 18 months. This study was approved by the East of England-Cambridge Central Research Ethics Committee and the Health Research Authority (reference number 16/EE/0370). We aim to publish the outputs of this work in international peer-reviewed journals compliant with an Open Access policy. NCT03412357.