Predictive value of RIFLE classification on prognosis of critically ill patients with acute kidney injury treated with continuous renal replacement therapy.
The optimal timing to start continuous renal replacement therapy (CRRT) for acute kidney injury (AKI) patients has not been accurately established. The recently proposed risk, injury, failure, loss, end-stage kidney disease (RIFLE) criteria for diagnosis and classification of AKI may provide a method for clinicians to decide the "optimal timing" for starting CRRT under uniform guidelines. The present study aimed: (1) to analyze the correlation between RIFLE stage at the start of CRRT and 90-day survival rate after CRRT start, (2) to further investigate the correlation of RIFLE stage with the malignant kidney outcome in the 90-day survivors, and (3) to determine the influence of the timing of CRRT defined by RIFLE classification on the 90-day survival and malignant kidney outcome in 90-day survivors. A retrospective cohort analysis was performed on the data of 106 critically ill patients with AKI, treated with CRRT during a 6-year period in a university affiliated surgical intensive care unit (SICU). Information such as sex, age, RIFLE stage, sepsis, sepsis-related organ failure assessment (SOFA) score, number of organ failures before CRRT, CRRT time during SICU, survival, and kidney outcome conditions at 90 days after CRRT start was collected. According to their baseline severity of AKI at the start of CRRT, the patients were assigned to three groups according to the increasing severity of RIFLE stages: RIFLE-R (risk of renal dysfunction, R), RIFLE-I (injury to the kidney, I) and RIFLE-F (failure of kidney function, F) using RIFLE criteria. The malignant kidney outcome was classified as RIFLE-L (loss of kidney function, L) or RIFLE-E (end-stage kidney disease, E) using RIFLE criteria. The correlation between RIFLE stage and 90-day survival rate was analyzed among these three RIFLE-categorized groups. Additionally, the association between RIFLE stage and the malignant kidney outcome (RIFLE-L + RIFLF-E) in the 90-day survivors was analyzed. Fifty-three of the overall 106 patients survived to 90 days after the start of CRRT. There were 16, 22 and 68 patients in RIFLE-R, RIFLE-I and RIFLE-F groups respectively with corresponding 90-day survival rate of 75.0% (12/16), 63.6% (14/22) and 39.7% (27/68) (P < 0.01, compared among groups). The percentage of the malignant kidney outcome of 90-day survivors in the RIFLE-R, RIFLE-I, and RIFLE-F groups was 16.7% (2/12), 21.4% (3/14) and 55.6% (15/27), respectively (P for trend < 0.01). After adjustment for other baseline risk factors, the relative risk (RR) for the 90-day mortality significantly increased with baseline RIFLE stage. Patients in RIFLE-F had a higher RR of 1.96 (95% confidence interval (CI): 1.06 - 3.62) than patients in RIFLE-I (RR: 1.09, 95% CI: 0.55 - 2.15) compared with patients in RIFLE-R (P for trend < 0.01). Similarly, baseline RIFLE stage also significantly correlated with the odds ratio (OR) for the malignant kidney outcome in 90-day survivors (P for trend < 0.05). Ninety-day survivors in the RIFLE-F group had a borderline significantly highest OR of 6.88 (95% CI: 0.85 - 55.67). The RIFLE classification may be used to predict 90-day survival after starting CRRT and the malignant kidney outcome of 90-day survivors in the critically ill patients with AKI treated with CRRT. Starting CRRT prior to RIFLE-F stage may be the optimal timing. Prospective, multi-center, randomized controlled trials are needed to confirm its predictive value in these patients.


The normal organization of the lateral posterior nucleus in the golden hamster and its reorganization after neonatal superior colliculus lesions.
We have studied the normal organization of the hamster lateral posterior nucleus and its reorganization after neonatal superior colliculus lesions. First, we divided the lateral posterior nucleus into rostrolateral, rostromedial and caudal subdivisions and determined the normal distributions of terminals contributed to each division by the ipsilateral and contralateral superior colliculi, the ipsilateral posterior neocortex and the contralateral retina. Since the rostrolateral subdivision receives most of the projections from the ipsilateral superior colliculus, our studies concentrated on this region. The rostrolateral subdivision contains synaptic clusters formed primarily by medium-sized M-terminals synapsing around a central dendrite. Electron microscopic observations showed that the majority of M-terminals are from the ipsilateral colliculus, although a few are contributed by the contralateral colliculus and retina. Therefore, after an ipsilateral neonatal colliculus lesion the synaptic clusters must develop in the absence of their major input. The next step was to examine the distributions of the remaining afferents to the rostrolateral subdivision in adult animals which had received ipsilateral neonatal colliculus lesions. In the cases, the normally restricted projection fields of the contralateral colliculus and the retina expand until they share a border in the rostrolateral subdivision. In contrast the cortical projection, which normally extends throughout the lateral posterior nucleus, is reduced in the region containing retinal terminals. At the ultrastructural level, we found morphologically normal synaptic clusters and showed the M-terminals now occupying the clusters are contributed by the remaining colliculus and the retina. The results suggested that the afferents to the lateral posterior nucleus normally compete for synaptic space and that this competition continues after a neonatal colliculus lesion. In our final experiments, we performed various combinations of neonatal lesions (bilateral superior colliculus, superior colliculus and retina, superior colliculus and cortex) and found that the remaining afferent expand their terminal fields still further in the absence of two inputs.


Digestibility of dietary fiber components in vegetarian men.
Digestibility of fiber components namely neutral detergent fiber (total content of cellwall) cellulose, hemicellulose and lignin are estimated in 14 healthy vegetarian men during adlibitum feeding and at 3 energy levels namely 2526, 2868 and 3290 kcals/day. Values of digestibility for adlibitum experiments were 34.17 +/- 2.3 for neutral detergent fiber (NDF), 30.1 +/- 3.9 for cellulose and 53.4 +/- 3.0 for hemicellulose and 8.1 +/- 2.6 for lignin. There was a considerable variability in digestibility of fiber components between individuals.


Functional expression of a vertebrate inwardly rectifying K+ channel in yeast.
We describe the expression of gpIRK1, an inwardly rectifying K+ channel obtained from guinea pig cardiac cDNA. gpIRK1 is a homologue of the mouse IRK1 channel identified in macrophage cells. Expression of gpIRK1 in Xenopus oocytes produces inwardly rectifying K+ current, similar to the cardiac inward rectifier current IK1. This current is blocked by external Ba2+ and Cs+. Plasmids containing the gpIRK1 coding region under the transcriptional control of constitutive (PGK) or inducible (GAL) promoters were constructed for expression in Saccharomyces cerevisiae. Several observations suggest that gpIRK1 forms functional ion channels when expressed in yeast. gpIRK1 complements a trk1 delta trk2 delta strain, which is defective in potassium uptake. Expression of gpIRK1 in this mutant restores growth on low potassium media. Growth dependent on gpIRK1 is inhibited by external Cs+. The strain expressing gpIRK1 provides a versatile genetic system for studying the assembly and composition of inwardly rectifying K+ channels.


Malondialdehyde and glutathione production in isolated perfused human and rat hearts.
A number of studies show the relation between oxygen-derived free radicals and cardiac ischemia/reperfusion injury. However, little is known about oxidative stress in the human heart, which can be measured by oxidized glutathione (GSSG) and malondialdehyde (MDA) formation. Furthermore, data on MDA production by rat hearts are controversial, possibly because of the use of the aspecific thiobarbituric acid assay. Therefore, GSSG and MDA were measured, with colorimetric and high-performance liquid chromatographic assays, respectively, in buffer-perfused explanted human hearts and normal rat hearts made temporarily ischemic. Human hearts received cardioplegia; rat hearts were studied in a control and an ischemic group with or without cardioplegia. Baseline GSSG release was < 0.01 nmol.min-1.g wet wt-1 in both species. During reperfusion, GSSG release from human hearts and from ischemic and cardioplegic/ischemic rat hearts peaked at 0.24 +/- 0.12, 1.1 +/- 0.4, and 0.19 +/- 0.04 nmol.min-1.g-1, respectively. MDA was undetectable (< 0.02 nmol.min-1.g-1) in the effluent of both species and in human hearts (< 4 nmol/g protein). Rat heart reduced glutathione levels decreased 32% as a consequence of cardioplegia and ischemia. Cardioplegia induced a 41% (P = .08) decrease in rat heart MDA content, whereas cumene hydroperoxide increased it 3.6 times (P < .01). Thus, after ischemia human and rat hearts release GSSG, indicating that oxidative stress has occurred. Apparently, lipid peroxidation takes place in normal rat hearts, decreases after cardioplegia, but does not increase after ischemia/reperfusion. Human hearts lack MDA under normoxic and ischemic conditions. This novel finding seems to reflect a low MDA-forming potential in both situations.(ABSTRACT TRUNCATED AT 250 WORDS)


Long-term clinical outcome after fractional flow reserve-guided percutaneous coronary intervention in patients with multivessel disease.
In the present study, we analyzed the clinical outcome of patients with multivessel coronary artery disease in whom at least one vessel was treated by percutaneous coronary intervention (PCI) and at least one other vessel was deferred on the basis of fractional flow reserve (FFR) measurements during the same session. Myocardial FFR is an established tool for assessing the severity of epicardial stenoses. It has been shown that it is safe to defer an intervention in single vessel disease patients when FFR >0.75. One hundred two patients (66 +/- 10 years) with multivessel coronary artery disease were included in the study. In all patients, PCI of at least two vessels was contemplated. Yet in all of them at least one vessel was treated by PCI, whereas at least one other vessel was deferred based on an FFR >0.75. Major adverse cardiac events (MACE) were recorded during an average follow-up of 29 +/- 18 months. In 102 patients, 113 coronary arteries underwent PCI. In these arteries FFR was 0.57 +/- 0.13 and mean diameter stenosis was 68 +/- 14%. One hundred twenty-seven coronary arteries had an FFR >0.75 and PCI was deferred. In these arteries FFR was 0.86 +/- 0.06 and mean diameter stenosis was 47 +/- 12%. No death occurred during the follow-up. A MACE occurred in 9% and 13% of patients after 12 and 36 months, respectively. These MACE were related to 22 (9.2%) arteries. Among them, 8 (6.3%) MACE were related to one of the initially deferred vessels, whereas 14 (12.3%) MACE were related to one of the initially treated coronary artery. In patients with multivessel disease, PCI of hemodynamically non-significant stenoses can be safely deferred, even if initially planned on the basis of the angiogram.


Identifying on admission patients likely to develop acute kidney injury in hospital.
The incidence of Acute Kidney Injury (AKI) continues to increase in the UK, with associated mortality rates remaining significant. Approximately one fifth of hospital admissions are associated with AKI and approximately a third of patients with AKI in hospital develop AKI during their time in hospital. A fifth of these cases are considered avoidable. Early risk detection remains key to decreasing AKI in hospitals, where sub-optimal care was noted for half of patients who developed AKI. Electronic anonymised data for adults admitted into the Royal Cornwall Hospitals Trust (RCHT) between 18th March and 31st December 2015 was trimmed to that collected within the first 24 h of hospitalisation. These datasets were split according to three separate time periods: data used for training the Takagi-Sugeno Fuzzy Logic Systems (FLS) and the multivariable logistic regression (MLR) models; data used for testing; and data from a later patient spell used for validation. Three fuzzy logic models and three MLR models were developed to link characteristics of patients diagnosed with a maximum stage AKI within 7 days of admission: the first models to identify any AKI Stage (FLS I, MLR I), the second for patterns of AKI Stage 2 or 3 (FLS II, MLR II), and the third to identify AKI Stage 3 (FLS III, MLR III). Model accuracy is expressed by area under the curve (AUC). Accuracy for each model during internal validation was: FLS I and MLR I (AUC 0.70, 95% CI: 0.64-0.77); FLS II (AUC 0.77, 95% CI: 0.69-0.85) and MLR II (AUC 0.74, 95% CI: 0.65-0.83); FLS III and MLR III (AUC 0.95, 95% CI: 0.92-0.98). FLS II and FLS III (and the respective MLR models) can identify with a high level of accuracy patients at high risk of developing AKI in hospital. These two models cannot be properly assessed against prior studies as this is the first attempt at quantifying the risk of developing specific Stages of AKI for a broad cohort of both medical and surgical inpatients. FLS I and MLR I performance is comparable to other existing models.


Have renal dietitians successfully implemented evidence-based guidelines into practice? A survey of dietitians across Australia and New Zealand.
Successful implementation of evidence-based practice (EBP) guidelines has been shown to improve the nutrition status of dialysis patients. This study aimed to establish use of EBP guidelines and implementation of key recommendations for nutrition assessment of dialysis patients, as well as to identify barriers and enablers associated with EBP guideline adherence. A survey of nutrition assessment practices and barriers to implementation of EBP guidelines was developed and piloted. The survey measured implementation of guidelines regarding frequency of nutrition assessment and use of the subjective global assessment (SGA) to diagnose malnutrition. Barriers to guideline implementation were measured using agreement with statements rated on a Likert scale. Data were summarized as counts and percentages and analyzed using chi-squared tests of association, with P < .05 indicating statistical significance. The survey targeted specialist renal dietitians across Australian and New Zealand. Sixty-five renal dietitians from Australia and New Zealand responded to the survey. Most were females (89%, n = 58 of 65), aged <35 years (72%, n = 47 of 65), with one-third (n = 22 of 65) working in renal dietetics for longer than 4 years. Nearly all participants (n = 62 of 65) reported routinely using EBP guidelines; however, only 55% and 66% indicated they had successfully implemented the guidelines regarding minimum 6-monthly nutrition assessment of dialysis patients (n = 36 of 65) and use of the SGA (n = 43 of 65), respectively. Barriers related to time, skills/self-efficacy, and an inefficient referral system were related to lower rates of guideline implementation. These findings indicate an evidence-practice gap in the nutritional management of dialysis patients. A standardized approach to EBP guideline implementation including structured 6-monthly nutrition assessment of dialysis patients and group training for use of the SGA tool may assist in closing this evidence-practice gap.


Management of failed UKA to TKA: conventional versus robotic-assisted conversion technique.
Failure of unicompartmental knee arthroplasty (UKA) is a distressing and technically challenging complication. Conventional conversion techniques (CCT) with rods and jigs have produced varying results. A robotic-assisted conversion technique (RCT) is an unexplored, though possibly advantageous, alternative. We compare our reconstructive outcomes between conventional and robotic methods in the management of failed UKA. Thirty-four patients with a failed UKA were retrospectively reviewed. Patients underwent conversion total knee arthroplasty (TKA) with either a CCT or RCT. Seventeen patients were included in each group. All procedures were done by a single surgeon at a single institution, with a mean time to follow-up of 3.6 years (range, 1 to 12). The primary outcome measures were the need for augments and polyethylene thickness. Secondary outcome measures were complications, need for revision, estimated blood loss (EBL), length of stay, and operative time. The mean polyethylene thickness was 12 mm (range, 9 to 15) in the CCT group and 10 mm (range, 9 to 14) in the RCT groups, with no statistical difference between the two groups (P = 0.07). A statistically significant difference, however, was present in the use of augments. In the CCT group, five out of 17 knees required augments, whereas none of the 17 knees in the RCT group required augments (P = 0.04). Procedurally, robotic-assisted surgery progressed uneventfully, even with metal artifact noted on the preoperative computerized tomography (CT) scans. Computer mapping of the residual bone surface after implant removal was a helpful guide in minimizing resection depth. No further revisions or reoperations were performed in either group. Robotic-assisted conversion TKA is technically feasible and potentially advantageous. In the absence of normal anatomic landmarks to guide conventional methods, the preoperative CT scans were unexpectedly helpful in establishing mechanical alignment and resection depth. In this limited series, RCT does not seem to be inferior to CCT. Further investigation of outcomes is warranted.


Changes in bacterial profile during amebiasis: demonstration of anaerobic bacteria in ALA pus samples.
Little is known about the changes in gut resident flora during amebic colitis and amebic liver abscess (ALA) caused by Entamoeba histolytica infection. Fecal samples from ALA patients, from healthy E. histolytica negative and positive (asymptomatic) individuals, and from pre- and post-metronidazole-treated healthy volunteers and pus samples from ALA patients were tested for the presence of various bacterial genera using 16S rRNA-based primers. Statistically significant reduction in Lactobacillus due to E. histolytica infection was observed in asymptomatic individuals and ALA patients. On the other hand, reduction in Bacteroides, Bifidobacterium, and Clostridium in the same samples was due to metronidazole treatment. Two anaerobic genera, viz. Bacteroides and Peptostreptococcus, were detected in ALA pus samples, and this observation is unprecedented. In addition, PCR revealed metronidazole resistance genes in fecal and pus samples of metronidazole-treated individuals. Re-examination of the ameba-bacterium relationship in amebiasis is suggested.


Minimally invasive foramen magnum durectomy and obexostomy for treatment of craniocervical junction-related syringomyelia in adults: case series and midterm follow-up.
Craniocervical junction-related syringomyelia (CCJS) is the most common form of syringomyelia. Approximately 30% of patients treated with foramen magnum decompression (FMD) will show persistence, recurrence, or progression of the syrinx. The authors present a pilot study with a new minimally invasive surgery technique targeting the pathophysiology of CCJS in adult patients. The authors retrospectively analyzed the clinical and radiological features of a consecutive series of patients treated for CCJS. An FMD and FM durectomy were performed through a 1.5- to 2-cm skin incision. Then arachnoid adhesions were cleared, creating a permanent communication from the fourth ventricle to the new paraspinal extradural cavity (obexostomy) and with the spinal subarachnoid space. The hypothesis was that the new CSF pouch acts like a pressure leak, interrupting the CCJS pathogenesis. Twenty-four patients (13 female, 21-61 years old) were treated between 2014 and 2018. The etiology of CCJS was Chiari malformation type I (CM-I) in 20 patients (83.3%), Chiari malformation type 0 (CM-0) in 2 patients (8.3%), and CCJ arachnoiditis in 2 patients (8.3%). Two patients underwent reoperations after failed FMD for CM-I at other institutions. No major surgical complication occurred. One patient had postoperative meningitis with no CSF fistula. On postoperative MRI, shrinkage of the syrinx was seen in all patients. No patients experienced recurrence of the CCJS. No patient required a subsequent operation. The mean duration of surgery was 72 ± 11 minutes (mean ± SD), and blood loss was 35-80 ml (mean 51 ml). Follow-up ranged from 12 to 58 months. The average overall improvement in modified Japanese Orthopaedic Association scores was 10% (p < 0.001). The Odom scale showed that 19 patients (79.1%) were satisfied, 4 (16.7%) remained the same, and 1 (4.2%) reported a poor outcome. All patients experienced postoperative improvement in perception of quality of life (p < 0.001). Minimally invasive FM durectomy and obexostomy is a safe and effective treatment for CCJS and for patients who have not responded to other treatment.


Long-Term Engraftment Promotes Differentiation of Alveolar Epithelial Cells from Human Embryonic Stem Cell Derived Lung Organoids.
Human embryonic stem cell (hESC) derived 3D human lung organoids (HLOs) provide a promising model to study human lung development and disease. HLOs containing proximal or/and immature distal airway epithelial cells have been successfully generated in vitro, such as early staged alveolar type 2 (AT2) cells (SPC+/SOX9+) and immature alveolar type 1 (AT1) cells (HOPX+/SOX9+). When HLOs were transplanted into immunocompromised mice for further differentiation in vivo, only few distal epithelial cells could be observed. In this study, we transplanted different stages of HLOs into immunocompromised mice to assess whether HLOs could expand and mature in vivo. We found that short-term transplanted HLOs contained lung progenitor cells (NKX2.1+, SOX9+, and P63+), but not SPC+ AT2 cells or AQP5+ AT1 cells. Meanwhile, long-term engrafted HLOs could differentiate into lung distal bipotent progenitor cells (PDPN+/SPC+/SOX9+), AT2 cells (SPC+, SPB+), and immature AT1 cells (PDPN+, AQP5-). However, HLOs at late in vitro stage turned into mature AT1-like cells (AQP5+/SPB-/SOX9-) in vivo. Immunofluorescence staining and transmission electron microscopy (TEM) results revealed that transplanted HLOs contained mesenchymal cells (collagen I+), vasculature (ACTA2+), neuroendocrine-like cells (PGP9.5+), and nerve fiber structures (myelin sheath structure). Together, these data reveal that hESC-derived HLOs would be useful for human lung development modeling, and transplanted HLOs could mimic lung organ-like structures in vivo by possessing vascular network and neuronal network.


Origin of repulsive force and structure/dynamics of interfacial water in OEG-protein interactions: a molecular simulation study.
Molecular simulations were performed to investigate the origin of the strong repulsive force acting on a protein as the protein approaches an oligo (ethylene glycol) self-assembled monolayer (OEG-SAM) surface. Since the repulsive force is mainly generated from water molecules, the force from the water molecules near the surface was calculated layer by layer to further identify the molecular origin of the repulsive force. Results show that the strong repulsive force acting on the protein near the OEG-SAM surface is dominantly generated by the interfacial water molecules located between the OEG-SAM surface and lysozyme. A hydroxyl-terminated SAM (OH-SAM) surface was used for comparison. No significant repulsive force was observed from the water molecules between the protein and OH-SAM surface. Further studies show that the dipole distribution of the interfacial water molecules is significantly affected by the OEG-SAM surface, as opposed to the negligible impact from the OH-SAM surface. The interfacial water molecules above the OEG-SAM surface stay longer and reorient more slowly than those above the OH-SAM surface. These results from this work support the hypothesis that the OEG-SAM surface interacts strongly with interfacial water molecules and creates a stable hydration layer that prevents proteins from adsorbing to the surface.


Neuropsychiatric symptoms in past users of sheep dip and other pesticides.
To explore the prevalence and pattern of neuropsychiatric symptoms in past users of sheep dip and other pesticides. From a postal survey of men born between 1933 and 1977 and resident in three rural areas of England and Wales (response rate 31%), data were obtained on lifetime history of work with pesticides, neurological symptoms in the past month, current mental health and tendency to be troubled by non-neurological somatic symptoms (summarised as a somatising tendency score). Risk factors for current neuropsychiatric symptoms were assessed by modified Cox regression. Data were available for 9844 men, including 1913 who had worked with sheep dip, 832 with other insecticides but not sheep dip and 990 with other pesticides but never with sheep dip or insecticides. Neurological symptoms were consistently 20-60% more common in past users of sheep dip than in men who had never worked with pesticides, but their prevalence was also higher in men who had worked only with pesticides other than sheep dip or insecticides. They clustered strongly within individuals, but this clustering was not specific to men who had worked with sheep dip. Reporting of three or more neurological symptoms was associated with somatising tendency (prevalence ratio (PR) 15.0, 95% CI 11.4 to 19.5, for the highest vs the lowest category of somatisation) and was more common in users of sheep dip (PR 1.3, 95% CI 1.0 to 1.6), other insecticides (PR 1.4, 95% CI 1.0 to 1.8) and other pesticides (PR 1.3, 95% CI 1.0 to 1.7) than in non-users. Among users of sheep dip, prevalence was higher in men who had dipped most often, but not in those who had worked with sheep dip concentrate. Past use of pesticides was not associated with current anxiety or depression. Neurological symptoms are more common in men who have worked with sheep dip, but the association is not specific to sheep dip or insecticides. A toxic cause for the excess cannot be ruled out, but several features of our observations suggest that psychological mechanisms have a role.


Current Progress in EBV-Associated B-Cell Lymphomas.
Epstein-Barr virus (EBV) was the first human tumor virus discovered more than 50 years ago. EBV-associated lymphomagenesis is still a significant viral-associated disease as it involves a diverse range of pathologies, especially B-cell lymphomas. Recent development of high-throughput next-generation sequencing technologies and in vivo mouse models have significantly promoted our understanding of the fundamental molecular mechanisms which drive these cancers and allowed for the development of therapeutic intervention strategies. This review will highlight the current advances in EBV-associated B-cell lymphomas, focusing on transcriptional regulation, chromosome aberrations, in vivo studies of EBV-mediated lymphomagenesis, as well as the treatment strategies to target viral-associated lymphomas.


Changes on microsatellites of expressed sequence tag of sugarcane (Saccharum spp) during vegetative propagation.
The reduction in sugarcane productivity in subsequent cutting stages may be related to a gradual decrease of the allele number and mean observed heterozygosity (HO) in the sugarcane ratoon. This hypothesis was tested assessing the number of alleles and HO values in 10 expressed sequence tag microsatellites (Est-SSR loci) of the sugarcane varieties RB72454 and RB867515 in different cutting stages. Changes of allele numbers in samples of different cutting stages were observed in seven and six EstSSR loci of the RB72454 and RB867515 varieties, respectively. Reduction of allele numbers was observed in the samples collected in the fourth and sixth cutting stages of the RB72454 variety. In contrast, an increase of the allele numbers was detected in the samples collected on fourth, sixth, and seventh cutting stages of the RB867515 variety. Unchanged allele numbers were observed only in EstB41, EstC84, and EstB130 loci of the RB72454 variety, and EstB41, EstC67, EstA68, and EstB130 loci of the RB867515 variety. The variety RB867515 has lower polymorphism and values of HO than the RB72454 variety in different stages of cutting. At molecular level, in Est-SSR loci, the RB72454 variety showed higher changes in subsequent stages of cutting than RB867515. The similarities and divergences at molecular level between varieties RB72454 and RB867515 observed in the 10 Est-SSR loci during subsequent cutting stages can not explain the reduced productivity frequently observed after subsequent cutting stages but showed that phenotypic and physiological changes after each cutting stage are also accompanied by changes at genomic level.


Elemental and molecular profiling of licit, illicit, and niche tobacco.
The recognition of differences between regulated large-scale mass manufactured products and the uncontrolled cultivation of tobaccos for illicit purposes plays a significant role within identification of provenance. This research highlights X-ray fluorescence and Fourier transform infrared spectroscopy as useful analytical techniques for the rapid identification of tobacco samples of unknown provenance. Identification of key discriminative features within each technique allowed for the development of typical characteristic profiles for each type of tobacco. Analysis using X-ray fluorescence highlights chlorine, potassium, calcium and iron as key elemental indicators of tobacco provenance. Significant levels of chlorine seen within Snüs samples prompted attempts to visualise chlorine containing regions and structures within the sample. Scanning electron microscopy images showed crystalline structures visible within the Snüs tobacco, structures which Energy dispersive X-ray spectroscopy qualitatively confirmed to contain chlorine. Chloride levels within Snüs samples were quantified using ion chromatography with levels found to range between 0.87mgmL(-1) and 1.28mg. Additionally, FTIR indicated that absorbances attributed to carbonyl stretching at 1050-1150cm(-1), alkane bending at 1350-1480cm(-1) and amide I stretching at 1600-1700cm(-1) highlighting a spectral fingerprint region that allowed for the clear differentiation between different types of tobaccos using PCA analysis, but was limited by differentiation between provenance of cigarettes and hand rolled tobacco. X-ray fluorescence and Fourier transform infrared spectroscopy yielded different information with regards tobacco discrimination and provenance, however both methods overall analysis time and cost reduced indicating usefulness as potential handheld analytical techniques in the field.


Does primary medical practitioner involvement with a specialist team improve patient outcomes? A systematic review.
Patients with chronic or complex medical or psychiatric conditions are treated by many practioners, including general practitioners (GPs). Formal liaison between primary and specialist is often assumed to offer benefits to patients. The aim of this study was to assess the efficacy of formal liaison of GPs with specialist service providers on patient health outcomes, by conducting a systematic review of the published literature in MEDLINE, EMBASE, PsychINFO, CINAHL and Cochrane Library databases using the following search terms: 'family physician': synonyms of 'patient care planning', 'patient discharge' and 'patient care team'; and synonyms of 'randomised controlled trials'. Seven studies were identified, involving 963 subjects and 899 controls. Most health outcomes were unchanged, although some physical and functional health outcomes were improved by formal liaison between GPs and specialist services, particularly among chronic mental illness patients. Some health outcomes worsened during the intervention. Patient retention rates within treatment programmes improved with GP involvement, as did patient satisfaction. Doctor (GP and specialist) behaviour changed with reports of more rational use of resources and diagnostic tests, improved clinical skills, more frequent use of appropriate treatment strategies, and more frequent clinical behaviours designed to detect disease complications. Cost effectiveness could not be determined. In conclusion, formal liaison between GPs and specialist services leaves most physical health outcomes unchanged, but improves functional outcomes in chronically mentally ill patients. It may confer modest long-term health benefits through improvements in patient concordance with treatment programmes and more effective clinical practice.


Rapid effects of deep brain stimulation reactivation on symptoms and neuroendocrine parameters in obsessive-compulsive disorder.
Improvement of obsessions and compulsions by deep brain stimulation (DBS) for obsessive-compulsive disorder (OCD) is often preceded by a rapid and transient mood elevation (hypomania). In a previous study we showed that improvement of mood by DBS for OCD is associated with a decreased activity of the hypothalamus-pituitary adrenal axis. The aim of our present study was to evaluate the time course of rapid clinical changes following DBS reactivation in more detail and to assess their association with additional neuroendocrine parameters. We included therapy-refractory OCD patients treated with DBS (>1 year) and performed a baseline assessment of symptoms, as well as plasma concentrations of thyroid-stimulating hormone (TSH), prolactin, growth hormone, copeptin and homovanillic acid. This was repeated after a 1-week DBS OFF condition. Next, we assessed the rapid effects of DBS reactivation by measuring psychiatric symptom changes using visual analog scales as well as repeated neuroendocrine measures after 30 min, 2 h and 6 h. OCD, anxiety and depressive symptoms markedly increased during the 1-week OFF condition and decreased again to a similar extent already 2 h after DBS reactivation. We found lower plasma prolactin (41% decrease, P=0.003) and TSH (39% decrease, P=0.003) levels during DBS OFF, which increased significantly already 30 min after DBS reactivation. The rapid and simultaneous increase in TSH and prolactin is likely to result from stimulation of hypothalamic thyrotropin-releasing hormone (TRH), which may underlie the commonly observed transient mood elevation following DBS.


The prevalence of psychosis in epilepsy; a systematic review and meta-analysis.
Epilepsy has long been considered to be a risk factor for psychosis. However there is a lack of consistency in findings across studies on the effect size of this risk which reflects methodological differences in studies and changing diagnostic classifications within neurology and psychiatry. The aim of this study was to assess the prevalence of psychosis in epilepsy and to estimate the risk of psychosis among individuals with epilepsy compared with controls. A systematic review and meta-analysis was conducted of all published literature pertaining to prevalence rates of psychosis in epilepsy using electronic databases PUBMED, OVIDMEDLINE, PsychINFO and Embase from their inception until September 2010 with the following search terms: prevalence, incidence, rate, rates, psychosis, schizophrenia, schizophreniform illness, epilepsy, seizures, temporal lobe epilepsy. The literature search and search of reference lists yielded 215 papers. Of these, 58 (27%) had data relevant to the review and 157 were excluded following a more detailed assessment. 10% of the included studies were population based studies. The pooled odds ratio for risk of psychosis among people with epilepsy compared with controls was 7.8. The pooled estimate of prevalence of psychosis in epilepsy was found to be 5.6% (95% CI: 4.8-6.4). There was a high level of heterogeneity. The prevalence of psychosis in temporal lobe epilepsy was 7% (95% CI: 4.9-9.1). The prevalence of interictal psychosis in epilepsy was 5.2% (95% CI: 3.3-7.2). The prevalence of postictal psychosis in epilepsy was 2% (95% CI: 1.2-2.8). Our systematic review found that up to 6% of individuals with epilepsy have a co-morbid psychotic illness and that patients have an almost eight fold increased risk of psychosis. The prevalence rate of psychosis is higher in temporal lobe epilepsy (7%). We suggest that further investigation of this association could give clues to the aetiology of psychosis.


Cognitive factors maintaining persecutory delusions in psychosis: the contribution of depression.
Depression is common in people with schizophrenia, but how it might directly contribute to the persistence of psychotic symptoms has rarely been tested. The key aim of the present study was to test whether depression and associated cognitive processes predict the maintenance of persecutory delusions. Three groups of participants were tested at baseline: 60 patients with persecutory delusions in the context of a schizophrenia spectrum diagnosis, 30 patients with depression, and 30 nonclinical controls. They completed interviewer and self-report assessments of depression and paranoia, and measures of six cognitive factors (schematic beliefs, experiential avoidance, autobiographical memory, problem solving, rumination, worry style). The patients with persecutory delusions were then assessed again, six months later. It was found that 50% of the patients with persecutory delusions met diagnostic criteria for major depression. Cognitive processes found to be associated with depression across the groups were negative schematic beliefs about the self, experiential avoidance and rumination, but not autobiographical memory or problem solving. The severity of initial depression in patients with persecutory delusions predicted the persistence of paranoia over six months. A number of cognitive factors also predicted the persistence of persecutory delusions, including negative schematic beliefs about the self, worry, and problem-solving difficulties. In conclusion, depression is common in patients with current persecutory delusions, and it shows similar cognitive features to major depressive disorder. The results of this study indicate that depression and related processes may contribute to the maintenance of paranoia. Trials are warranted of depression-related therapeutic techniques for people with persecutory delusions.


Functional characteristics and sites of gene expression of the alpha 1, beta 1, gamma 2-isoform of the rat GABAA receptor.
GABAA receptors, the major synaptic targets for the neurotransmitter GABA, constitute gated chloride channels. By their allosteric, drug-induced modulation, they serve as control elements for the regulation of anxiety, vigilance, and epileptiform activity. The structural requirements of fully functional GABAA receptors in the mammalian brain have remained elusive so far. We report here on the cloning of the gamma 2-subunit cDNA of rat brain and its functional analysis by coexpression with the alpha 1- and beta 1-subunits in Xenopus oocytes, and on the sites of gene expression of the 3 subunits in the rat brain. The recombinant receptor displayed GABA-inducible currents (Imax = 6 microA; Ka = 75 microM) which were allosterically modulated by benzodiazepine receptor ligands (enhancement and inhibition by diazepam and methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate, respectively). In the absence of GABA, pentobarbital elicited a maximal current amplitude similar to that of GABA. A minor population of channels is expressed which is open in the absence of GABA or pentobarbital. Mapping subunit gene expression by in situ hybridization histochemistry suggests that the alpha 1-, beta 1-, and gamma 2-subunits are likely receptor constituents in some neuronal populations, e.g., mitral cells of the olfactory bulb, pyramidal cells of the hippocampus, and granule cells of the dentate gyrus and cerebellum.


Discriminant value of thyroid function tests.
Different thyroid function tests permitted a final classification of 204 consecutive patients with suspected thyroid disorders into three populations (thyrotoxic, euthyroid, and hypothyroid). Linear discriminant analysis was applied to all test results (10 variates) on adjacent population pairs. Two invitro tests (serum protein bound iodine (P.B.I.) and tri-iodothyronine (T-3) uptake values) gave good separation of thyrotoxic from euthyroid patients and fairly good distinction of hypothyroid patients. If a (131)I uptake figure was then added to the in-vitro results most patients (95.5%), including thse initially classified as equivocal, were correctly diagnosed. Other tests, including clinical questionnaires, were poor discriminants.Two new techniques of utilizing the test data were devised. Firstly, the data from the two in-vitro tests were also displayed graphically, and oblique boundary lines derived from the discriminant functions gave better separation of patients than previously used limits or mathematical expressions of "free thyroxine." Secondly, a nomogram incorporating the best four discriminants was designed as a diagnostic aid and proved to be the best means of interpreting the tests.Discriminant analysis of this kind can be used in the interpretation of diagnostic tests in any branch of medicine, and it allows the best use to be made of the available data.


The effects of parent-child relationships on later life mental health status in two national birth cohorts.
Abusive and neglectful parenting is an established determinant of adult mental illness, but longitudinal studies of the impact of less severe problems with parenting have yielded inconsistent findings. In the face of growing interest in mental health promotion, it is important to establish the impact of this potentially remediable risk factor. 8,405 participants in the 1958 UK birth cohort study, and 5,058 in the 1970 birth cohort study questionnaires relating to the quality of relationships with parents completed at age 16 years. 12-item General Health Questionnaire and the Malaise Inventory collected at age 42 years (1958 cohort) and 30 years (1970 cohort). Statistical methodology: logistic regression analyses adjusting for sex, social class and teenage mental health problems. 1958 cohort: relationships with both mother and father predicted mental health problems in adulthood; increasingly poor relationships were associated with increasing mental health problems at age 42 years. 1970 cohort: positive items derived from the Parental Bonding Instrument predicted reduced risk of mental health problems; negative aspects predicted increased risk at age 30 years. Odds of mental health problems were increased between 20 and 80% in fully adjusted models. Results support the hypothesis that problems with parent-child relationships that fall short of abuse and neglect play a part in determining adult mental health and suggest that interventions to support parenting now being implemented in many parts of the Western world may reduce the prevalence of mental illness in adulthood.


Record linkage is feasible with non-identifiable trauma and rehabilitation datasets.
1) Describe probabilistic linkage (PL) for road trauma and rehabilitation records in New South Wales (NSW) Australia. 2) Determine the accuracy of linkage for these records. Data were extracted from the NSW Trauma Registry for all road trauma admissions for the years 2009-2012 and from Australasian Rehabilitation Outcomes Centre for January 2009 to June 2013. PL was performed using: age; sex; residential postcode; and date of acute discharge = date of admission to rehabilitation. False matches were cases that linked but were not true matches; they were determined by manual review. Reasons for incomplete linkages were explored. The benefits and limitations of the linked study dataset are described. Of 3,256 road trauma records, 683 were matched to rehabilitation records. Using the field of 'discharge destination' from the trauma records, 265 patients with unmatched records were discharged to inpatient rehabilitation (missed matches). This gave an overall 72% linkage rate (or sensitivity) using PL. There were 16 cases of false matches, giving a specificity of 99%. It was feasible to use PL to link road trauma and rehabilitation datasets in the absence of identifiers. However, this needed to be combined with careful manual review before the linked dataset could be used to make inferences on trauma rehabilitation outcomes. PL may be a cost-effective way to capture inpatient rehabilitation outcomes of multi-trauma patients.


The Emergence of Chromosomally Located bla CTX-M-55 in Salmonella From Foodborne Animals in China.
The emergence and increase in prevalence of resistance to cephalosporins amongst isolates of Salmonella from food animals imposes a public health threat. The aim of the present study was to investigate the prevalence and characteristics of CTX-M-producing Salmonella isolates from raw meat and food animals. 27 of 152 (17.76%) Salmonella isolates were ESBL-positive including 21/70 (30%) from food animals and 6/82 (7.32%) from raw meat. CTX-M-55 was the most prevalent ESBL type observed (12/27, 44.44%). 7 of 12 CTX-M-55-positive Salmonella isolates were Salmonella Indiana, 2 were Salmonella Typhimurium, 2 were Salmonella Chester, and the remaining isolate was not typeable. Eight CTX-M-55-positive Salmonella isolates were highly resistant to fluoroquinolones (MICCIP = 64 ug/mL) and co-harbored aac(6')-Ib-cr and oqxAB. Most of the CTX-M-55 positive isolates (11/12) carried bla CTX-M-55 genes on the chromosome, with the remaining isolate carrying this gene on a transferable 280 kb IncHI2 plasmid. A chromosomal bla CTX-M-55 gene from one isolate transferred onto a 250 kb IncHI2 plasmid which was subsequently conjugated into recipient strain J53. PFGE and MLST profiles showed a wide range of strain types were carrying bla CTX-M-55. Our study demonstrates the emergence and prevalence of foodborne Salmonella harboring a chromosomally located bla CTX-M-55 in China. The co-existence of PMQR genes with bla CTX-M-55 in Salmonella isolates suggests co-selection and dissemination of resistance to both fluoroquinolones and cephalosporins in Salmonella via the food chain in China represents a public health concern.


Assessment of frontal lobe functions.
The authors describe methods for conducting a thorough assessment of functions subserved by the frontal lobes, employing both bedside and psychometric methods of assessing frontal subsystems. Qualitative or process aspects of frontal behavior observable from formal testing, interview, and social behavior are noted. It is argued that the skilled clinician must be guided by a knowledge of frontal lobe subsystems and their roles in determining specific types of abnormal behavior. The clinician will then be alert to changes in incidental behaviors that indicate frontal impairment, and bedside maneuvers can be designed to discriminate dysfunction. Given the complexity of the behaviors involved and the profound effects of maturation and aging on frontal functions, neuropsychological assessment can provide an invaluable tool for testing these functions.


Rare Detection of Antiviral Functions of Polyclonal IgA Isolated from Plasma and Breast Milk Compartments in Women Chronically Infected with HIV-1.
The humoral response to invading mucosal pathogens comprises multiple antibody isotypes derived from systemic and mucosal compartments. To understand the contribution of each antibody isotype/source to the mucosal humoral response, parallel investigation of the specificities and functions of antibodies within and across isotypes and compartments is required. The role of IgA against HIV-1 is complex, with studies supporting a protective role as well as a role for serum IgA in blocking effector functions. Thus, we explored the fine specificity and function of IgA in both plasma and mucosal secretions important to infant HIV-1 infection, i.e., breast milk. IgA and IgG were isolated from milk and plasma from 20 HIV-1-infected lactating Malawian women. HIV-1 binding specificities, neutralization potency, inhibition of virus-epithelial cell binding, and antibody-mediated phagocytosis were measured. Fine-specificity mapping showed IgA and IgG responses to multiple HIV-1 Env epitopes, including conformational V1/V2 and linear V2, V3, and constant region 5 (C5). Env IgA was heterogeneous between the milk and systemic compartments (Env IgA, τ = 0.00 to 0.63, P = 0.0046 to 1.00). Furthermore, IgA and IgG appeared compartmentalized as there was a lack of correlation between the specificities of Env-specific IgA and IgG (in milk, τ = -0.07 to 0.26, P = 0.35 to 0.83). IgA and IgG also differed in functions: while neutralization and phagocytosis were consistently mediated by milk and plasma IgG, they were rarely detected in IgA from both milk and plasma. Understanding the ontogeny of the divergent IgG and IgA antigen specificity repertoires and their effects on antibody function will inform vaccination approaches targeted toward mucosal pathogens.IMPORTANCE Antibodies within the mucosa are part of the first line of defense against mucosal pathogens. Evaluating mucosal antibody isotypes, specificities, and antiviral functions in relationship to the systemic antibody profile can provide insights into whether the antibody response is coordinated in response to mucosal pathogens. In a natural immunity cohort of HIV-infected lactating women, we mapped the fine specificity and function of IgA in breast milk and plasma and compared these with the autologous IgG responses. Antigen specificities and functions differed between IgG and IgA, with antiviral functions (neutralization and phagocytosis) predominantly mediated by the IgG fraction in both milk and plasma. Furthermore, the specificity of milk IgA differed from that of systemic IgA. Our data suggest that milk IgA and systemic IgA should be separately examined as potential correlates of risk. Preventive vaccines may need to employ different strategies to elicit functional antiviral immunity by both antibody isotypes in the mucosa.


Case Report: Typhoid Fever Complicated by Ileal Perforation in an Urban Slum of Dhaka, Bangladesh.
Intestinal perforation is one of the most dangerous complications of typhoid fever and demands urgent hospitalization, diagnosis, and surgical management to reduce morbidity and prevent mortality. Here, we report a case of typhoidal intestinal perforation in a 19 year-old young man detected by passive surveillance during a cluster-randomized trial with Vi-tetanus toxoid conjugate vaccine (Typhoid Vaccine Acceleration Consortium: TyVAC) in an urban slum area in Mirpur, Dhaka, Bangladesh. The patient presented with a high-grade fever, lower abdominal pain, and vomiting and was admitted to a healthcare facility. Physical examination and preoperative investigations of the patient suggested a presumptive diagnosis of intestinal perforation, and the patient was transferred to a tertiary-level hospital for surgical management. A positive blood culture, intraoperative findings, and histopathology of an intestinal biopsy confirmed ileal perforation due to typhoid fever. This case report highlights the need for prompt diagnosis and appropriate pre- and postoperative management of patients who appear with the symptoms of typhoidal intestinal perforation. This report further demonstrates the importance of systematic surveillance and proper evaluation to determine the true incidence rate of typhoid fever and intestinal perforation in Bangladesh.


Good practice in health care for migrants: views and experiences of care professionals in 16 European countries.
Health services across Europe provide health care for migrant patients every day. However, little systematic research has explored the views and experiences of health care professionals in different European countries. The aim of this study was to assess the difficulties professionals experience in their service when providing such care and what they consider constitutes good practice to overcome these problems or limit their negative impact on the quality of care. Structured interviews with open questions and case vignettes were conducted with health care professionals working in areas with high proportion of migrant populations in 16 countries. In each country, professionals in nine primary care practices, three accident and emergency hospital departments, and three community mental health services (total sample = 240) were interviewed about their views and experiences in providing care for migrant patients, i.e. from first generation immigrant populations. Answers were analysed using thematic content analysis. Eight types of problems and seven components of good practice were identified representing all statements in the interviews. The eight problems were: language barriers, difficulties in arranging care for migrants without health care coverage, social deprivation and traumatic experiences, lack of familiarity with the health care system, cultural differences, different understandings of illness and treatment, negative attitudes among staff and patients, and lack of access to medical history. The components of good practice to overcome these problems or limit their impact were: organisational flexibility with sufficient time and resources, good interpreting services, working with families and social services, cultural awareness of staff, educational programmes and information material for migrants, positive and stable relationships with staff, and clear guidelines on the care entitlements of different migrant groups. Problems and good care components were similar across the three types of services. Health care professionals in different services experience similar difficulties when providing care to migrants. They also have relatively consistent views on what constitutes good practice. The degree to which these components already are part of routine practice varies. Implementing good practice requires sufficient resources and organisational flexibility, positive attitudes, training for staff and the provision of information.


Biomechanical evaluation of polylactide absorbable screws used for syndesmosis injury repair.
Simulated syndesmosis injuries were created in 12 fresh-frozen, below-knee cadaver specimens. Six specimens were repaired with a 4.5 mm stainless steel screw, and six were repaired with a 4.5 mm polylactide screw. Three specimens of each group were tested in load to failure by axially loading with 1400 N and externally rotating to 90 degrees. Three specimens in each group underwent fatigue testing by axially loading with 700 N and applying 2.5 N-m of torque for 57,700 cycles. Radiographs and computed tomography scans were evaluated. None of the screws broke or failed. Similar load to failure was noted in polylactide and control groups. Fatigue testing revealed no significant change in stiffness. No significant screw damage was evident on radiographic or computed tomography evaluation. The data suggest that a polylactide screw has sufficient fatigue and failure strength to allow for healing of this injury in a clinical situation.


The effect of anesthetic technique on postoperative outcomes in hip fracture repair.
The impact of anesthetic choice on postoperative mortality and morbidity has not been determined with certainty. The authors evaluated the effect of type of anesthesia on postoperative mortality and morbidity in a retrospective cohort study of consecutive hip fracture patients, aged 60 yr or older, who underwent surgical repair at 20 US hospitals between 1983 and 1993. The primary outcome was defined as death within 30 days of the operative procedure. The secondary outcomes were postoperative 7-day mortality, postoperative myocardial infarction, postoperative pneumonia, postoperative congestive heart failure, and postoperative change in mental status. Numerous comorbid conditions were controlled for individually and by several comorbidity indices using logistic regression. General anesthesia was used in 6,206 patients (65.8%) and regional anesthesia in 3,219 patients (3,078 spinal anesthesia and 141 epidural anesthesia). The 30-day mortality rate in the general anesthesia group was 4.4%, compared with 5.4% in the regional anesthesia group (unadjusted odds ratio = 0.80; 95% confidence interval = 0.66-0.97). However, the adjusted odds ratio for general anesthesia increased to 1.08 (0.84-1.38). The adjusted odds ratios for general anesthesia versus regional anesthesia for the 7-day mortality was 0.90 (0.59-1.39) and for postoperative morbidity outcomes were as follows: myocardial infarction: adjusted odds ratio = 1.17 (0.80-1.70); congestive heart failure: adjusted odds ratio = 1.04 (0.80-1.36); pneumonia: adjusted odds ratio = 1.21 (0.87-1.68); postoperative change in mental status: adjusted odds ratio = 1.08 (0.95-1.22). The authors were unable to demonstrate that regional anesthesia was associated with better outcome than was general anesthesia in this large observational study of elderly patients with hip fracture. These results suggest that the type of anesthesia used should depend on factors other than any associated risks of mortality or morbidity.


¡Sí, Yo Puedo Vivir Sano con Diabetes! A Self-Management Randomized Controlled Pilot Trial for Low-Income Adults with Type 2 Diabetes in Mexico City.
Type 2 diabetes (T2D) is a worldwide epidemic and a leading cause of death in Mexico, with a prevalence of 15.9%, and >70% of diagnosed adults have poor glycemic control [glycated hemoglobin (HbA1c) >7.5%]. We developed a diabetes self-management education program contextualized to the study population, including dietary preferences, health literacy, and health system. We aimed to evaluate the efficacy of a self-management + text message program (¡Sí, Yo Puedo Vivir Sano con Diabetes!) on primary (HbA1c), and secondary behavioral (self-management), clinical, and psychosocial outcomes in adults with T2D in Mexico City. Participants were recruited at public primary healthcare centers (Seguro Popular), and randomly allocated to treatment (n = 26) or wait-list control groups (n = 21) with data collected at 3 and 6 mo. The program included 7 weekly sessions and 6 mo of daily text/picture messages. Descriptive statistics and a generalized linear mixed model with intent-to-treat analysis were calculated. Participants were 55.5 ± 8.8 y of age (mean ± SD), 68% female, 88.6% overweight/obese, and 57% lived in food-insecure households. Mean ± SD T2D duration was 11.9 ± 7.8 y and HbA1c was 9.2% ± 1.5%. There was 89% attendance at sessions and 6.4% attrition across both groups at 6 mo. Group-by-time effects were seen in self-monitoring of blood glucose (P < 0.01) and diabetes self-efficacy (P < 0.04); and a trend for lower HbA1c was seen in the intervention group at 6 mo (P = 0.11). Significant improvements in dietary behavior (P < 0.01) were demonstrated in the intervention group over time, but this did not reach statistical significance compared with the control group. The program was associated with clinically significant improvements in T2D self-management, self-efficacy, and HbA1c over time. Thus, T2D self-management skills, including diet, were improved in a vulnerable metropolitan population.This trial was registered at clinicaltrials.gov as NCT03159299.


Bleach gel: a simple agarose gel for analyzing RNA quality.
RNA-based applications requiring high-quality, non-degraded RNA are a foundational element of many research studies. As such, it is paramount that the integrity of experimental RNA is validated prior to cDNA synthesis or other downstream applications. In the absence of expensive equipment such as microfluidic electrophoretic devices, and as an alternative to the costly and time-consuming standard formaldehyde gel, RNA quality can be quickly analyzed by adding small amounts of commercial bleach to TAE buffer-based agarose gels prior to electrophoresis. In the presence of low concentrations of bleach, the secondary structure of RNA is denatured and potential contaminating RNases are destroyed. Because of this, the 'bleach gel' is a functional approach that addresses the need for an inexpensive and safe way to evaluate RNA integrity and will improve the ability of researchers to rapidly analyze RNA quality.


Effectiveness of school-based occupational therapy intervention on handwriting.
This study investigated the effects of school-based occupational therapy services on students' handwriting. Students 7 to 10 years of age with poor handwriting legibility who received direct occupational therapy services (n = 29) were compared with students who did not receive services (n = 9) on handwriting legibility and speed and associated performance components. Visual-motor, visual-perception, in-hand manipulation, and handwriting legibility and speed were measured at the beginning and end of the academic year. The intervention group received a mean of 16.4 sessions and 528 min of direct occupational therapy services during the school year. According to the therapists, visual-motor skills and handwriting practice were emphasized most in intervention. Students in the intervention group showed significant increases in in-hand manipulation and position in space scores. They also improved more in handwriting legibility scores than the students in the comparison group. Fifteen students in the intervention group demonstrated greater than 90% legibility at the end of the school year. On average, legibility increased by 14.2% in the students who received services and by 5.8% in the students who did not receive services. Speed increased slightly more in the students who did not receive services. Students who received occupational therapy services demonstrated improved letter legibility, but speed and numeral legibility did not demonstrate positive intervention effects.


NAFLD: Reporting Histologic Findings in Clinical Practice.
The role of liver biopsy in NASH has evolved along with the increased recognition of the significance of this disease, and the unmet medical need it presents. Drug development and clinical trials are rapidly growing, as are noninvasive tests for markers of steatosis, inflammation, injury, and fibrosis. Liver biopsy evaluation remains necessary for both drug development and clinical trials as the most specific means of diagnosis and patient identification for appropriate intervention. This White Paper, sponsored by the American Association for the Study of Liver Disease NASH Task Force, is a focused review of liver biopsy evaluation in fatty liver disease in subjects with presumed NAFLD for practicing clinical hepatologists and pathologists. The goal is to provide succinct and specific means for reporting the histopathologic elements of NASH, distinguishing NASH from nonalcoholic fatty liver without steatohepatitis, and from alcohol-associated steatohepatitis when possible. The discussion includes the special situations of NASH in advanced fibrosis or cirrhosis, and in the pediatric population. Finally, there is discussion of semiquantitative methods of evaluation of lesions of "disease activity" and fibrosis. Tables are presented for scoring and a suggested model for final reporting. Figures are presented to highlight the histopathologic elements of NASH.


Upholding science in health, safety and environmental risk assessments and regulations.
A public appeal has been advanced by a large group of scientists, concerned that science has been misused in attempting to quantify and regulate unmeasurable hazards and risks.1 The appeal recalls that science is unable to evaluate hazards that cannot be measured, and that science in such cases should not be invoked to justify risk assessments in health, safety and environmental regulations. The appeal also notes that most national and international statutes delineating the discretion of regulators are ambiguous about what rules of evidence ought to apply. Those statutes should be revised to ensure that the evidence for regulatory action is grounded on the standards of the scientific method, whenever feasible. When independent scientific evidence is not possible, policies and regulations should be informed by publicly debated trade-offs between socially desirable uses and social perceptions of affordable precaution. This article explores the premises, implications and actions supporting the appeal and its objectives.


Correlates of Level and Loss of Grip Strength in Later Life: Findings from the English Longitudinal Study of Ageing and the Hertfordshire Cohort Study.
Characterisation of grip strength (GS) using isometric dynamometry is central to the definition of sarcopenia. Determinants of low GS include: older age, shorter stature, low physical activity, poor nutrition, socioeconomic disadvantage and multimorbidity. Less is known about risk factors for accelerated loss of GS. We investigated determinants of level and 8-year loss of GS in 3703 men and women (aged 52-82 years) in the English Longitudinal Study of Ageing (ELSA). Four hundred and forty-one men and women (aged 59-71 years) who participated in a 10-year follow-up of the Hertfordshire Cohort Study (HCS) were used for replication. Variables were harmonised between cohorts. Change in GS was characterised using mixed-effects models in ELSA and a residual change approach in HCS and analysed for men and women combined. Men in ELSA and HCS had higher average levels of GS at baseline, and accelerated rates of loss, compared with women. In ELSA, older age, shorter stature and multimorbidity were correlated with lower level, and accelerated rate of loss, of GS in both sexes (accelerated loss of 0.04 (95% CI 0.00-0.08) standard deviation scores per additional morbidity after multivariable adjustment). Socioeconomic disadvantage, low level of physical activity and poorer self-reported health were also correlated with low GS level, but not loss rate, after multivariable adjustment. Analysis in HCS yielded similar results. Our results identify multimorbidity as a modifiable determinant of loss of muscle strength in later life, and raise the possibility that developmental influences may impact on rate of involutional decline in muscle strength.


Anthropogenic iodine-129 in seawater along a transect from the Norwegian coastal current to the North Pole.
Variation in the concentrations of iodine-129 (129I, T1/2=15.7 Myr), a low-level radioactive component of nuclear fuel waste, is documented in surface waters and depth profiles collected during 2001 along a transect from the Norwegian Coastal Current to the North Pole. The surface waters near the Norwegian coast are found to have 20 times higher 129I concentration than the surface waters of the Arctic Ocean. The depth profiles of 129I taken in the Arctic Ocean reveal a sharp decline in the concentration to a depth of about 300-500 m followed by a weaker gradient extending down to the bottom. A twofold increase in the 129I concentration is observed in the upper 1000 m since 1996. Based on known estimates of marine transient time from the release sources (the nuclear reprocessing facilities at La Hague, France, and Sellafield, UK), a doubling in the 129I inventory of the top 1000 m of the Arctic Ocean is expected to occur between the years 2001 and 2006. As 129I of polar mixed layer and Atlantic layer of the Arctic Ocean is ventilated by the East Greenland Current into the Nordic Seas and North Atlantic Ocean, further dispersal and increase of the isotope concentration in these regions will be encountered in the near future.


Etiological factors in space motion sickness.
We compared susceptibility to motion sickness during exposure to sudden-stop stimulation as a function of gravitoinertial force level. Our findings show that susceptibility is greatly enhanced, both with eyes-closed and eyes-open, for zero-g and 2-g conditions in parabolic flight compared with 1-g test conditions. The change in susceptibility is likely related to three factors: alterations in vestibulo-ocular function which result from variations in gravitoinertial force level (28,29); the altered pattern of otolithic activity resulting during variations in gravitoinertial force level; and the altered canal-otolith response synergies that result during exposure to gravitoinertial force levels greater or less than terrestrial levels. These factors are shown to be related to the etiology of space motion sickness and to the alterations in performance and vestibular function that are experienced by astronauts during reentry. An explanation is also proposed for the decrease in susceptibility to motion sickness exhibited by the Skylab astronauts inflight and for some period postflight during exposure to cross-coupled angular accelerations.


Consistent temperature coupling with thermal fluctuations of smooth particle hydrodynamics and molecular dynamics.
We propose a thermodynamically consistent and energy-conserving temperature coupling scheme between the atomistic and the continuum domain. The coupling scheme links the two domains using the DPDE (Dissipative Particle Dynamics at constant Energy) thermostat and is designed to handle strong temperature gradients across the atomistic/continuum domain interface. The fundamentally different definitions of temperature in the continuum and atomistic domain - internal energy and heat capacity versus particle velocity - are accounted for in a straightforward and conceptually intuitive way by the DPDE thermostat. We verify the here-proposed scheme using a fluid, which is simultaneously represented as a continuum using Smooth Particle Hydrodynamics, and as an atomistically resolved liquid using Molecular Dynamics. In the case of equilibrium contact between both domains, we show that the correct microscopic equilibrium properties of the atomistic fluid are obtained. As an example of a strong non-equilibrium situation, we consider the propagation of a steady shock-wave from the continuum domain into the atomistic domain, and show that the coupling scheme conserves both energy and shock-wave dynamics. To demonstrate the applicability of our scheme to real systems, we consider shock loading of a phospholipid bilayer immersed in water in a multi-scale simulation, an interesting topic of biological relevance.


Perioperative anesthesia management for pulmonary endarterectomy: Adopting an established European Protocol for the Asian Population.
Anesthesia for pulmonary endarterectomy (PEA) has always been one of the challenges of anesthesia. As one of the leading cardiothoracic institutions in Southeast Asia, our hospital has vast interest in this subject. A local multidisciplinary team was deployed to an expert center in the United Kingdom (UK), and the experience was then integrated to the care of our patients. We present a case series of ten patients undergoing anesthesia for PEA, a first for our institution, and discuss techniques as well as potential complications. Patients with chronic thromboembolic pulmonary hypertension were reviewed by a multidisciplinary team, and those who were suitable for surgical intervention subsequently underwent PEA. A total of ten patients were identified and operated on. The perioperative management and conduction of anesthesia for all patients followed a protocol adapted from the expert center in the UK, with revisions to cater to our Asian population. In the ten patients operated on, eight of them were successfully extubated on the first postoperative day. Apart from one incident of prolonged ventilator usage due to reperfusion lung injury and pneumonia, there were no major respiratory or hemodynamic complications. Certainly, six of the ten patients developed subdural hemorrhage after the commencement of enoxaparin, although none of them sustained any permanent neurological deficits. We have demonstrated that with careful planning and a well-outlined protocol, anesthesia for PEA in an Asian population can be achieved with favorable outcomes. Further fine-tuning of the protocol is still required based on local expertise.


Recurrent stroke risk and cerebral microbleed burden in ischemic stroke and TIA: A meta-analysis.
To determine associations between cerebral microbleed (CMB) burden with recurrent ischemic stroke (IS) and intracerebral hemorrhage (ICH) risk after IS or TIA. We identified prospective studies of patients with IS or TIA that investigated CMBs and stroke (ICH and IS) risk during ≥3 months follow-up. Authors provided aggregate summary-level data on stroke outcomes, with CMBs categorized according to burden (single, 2-4, and ≥5 CMBs) and distribution. We calculated absolute event rates and pooled risk ratios (RR) using random-effects meta-analysis. We included 5,068 patients from 15 studies. There were 115/1,284 (9.6%) recurrent IS events in patients with CMBs vs 212/3,781 (5.6%) in patients without CMBs (pooled RR 1.8 for CMBs vs no CMBs; 95% confidence interval [CI] 1.4-2.5). There were 49/1,142 (4.3%) ICH events in those with CMBs vs 17/2,912 (0.58%) in those without CMBs (pooled RR 6.3 for CMBs vs no CMBs; 95% CI 3.5-11.4). Increasing CMB burden increased the risk of IS (pooled RR [95% CI] 1.8 [1.0-3.1], 2.4 [1.3-4.4], and 2.7 [1.5-4.9] for 1 CMB, 2-4 CMBs, and ≥5 CMBs, respectively) and ICH (pooled RR [95% CI] 4.6 [1.9-10.7], 5.6 [2.4-13.3], and 14.1 [6.9-29.0] for 1 CMB, 2-4 CMBs, and ≥5 CMBs, respectively). CMBs are associated with increased stroke risk after IS or TIA. With increasing CMB burden (compared to no CMBs), the risk of ICH increases more steeply than that of IS. However, IS absolute event rates remain higher than ICH absolute event rates in all CMB burden categories.


Statistical challenges in the development and evaluation of marker-based clinical tests.
Exciting new technologies for assessing markers in human specimens are now available to evaluate unprecedented types and numbers of variations in DNA, RNA, proteins, or biological structures such as chromosomes. These markers, whether viewed individually, or collectively as a 'signature', have the potential to be useful for disease risk assessment, screening, early detection, prognosis, therapy selection, and monitoring for therapy effectiveness or disease recurrence. Successful translation from basic research findings to clinically useful test requires basic, translational, and regulatory sciences and a collaborative effort among individuals with varied types of expertise including laboratory scientists, technology developers, clinicians, statisticians, and bioinformaticians. The focus of this commentary is the many statistical challenges in translational marker research, specifically in the development and validation of marker-based tests that have clinical utility for therapeutic decision-making.


What Is Special about Face Recognition? Nineteen Experiments on a Person with Visual Object Agnosia and Dyslexia but Normal Face Recognition.
In order to study face recognition in relative isolation from visual processes that may also contribute to object recognition and reading, we investigated CK, a man with normal face recognition but with object agnosia and dyslexia caused by a closed-head injury. We administered recognition tests of up right faces, of family resemblance, of age-transformed faces, of caricatures, of cartoons, of inverted faces, and of face features, of disguised faces, of perceptually degraded faces, of fractured faces, of faces parts, and of faces whose parts were made of objects. We compared CK's performance with that of at least 12 control participants. We found that CK performed as well as controls as long as the face was upright and retained the configurational integrity among the internal facial features, the eyes, nose, and mouth. This held regardless of whether the face was disguised or degraded and whether the face was represented as a photo, a caricature, a cartoon, or a face composed of objects. In the last case, CK perceived the face but, unlike controls, was rarely aware that it was composed of objects. When the face, or just the internal features, were inverted or when the configurational gestalt was broken by fracturing the face or misaligning the top and bottom halves, CK's performance suffered far more than that of controls. We conclude that face recognition normally depends on two systems: (1) a holistic, face-specific system that is dependent on orientationspecific coding of second-order relational features (internal), which is intact in CK and (2) a part-based object-recognition system, which is damaged in CK and which contributes to face recognition when the face stimulus does not satisfy the domain-specific conditions needed to activate the face system.


Peptide engineered microcantilevers for selective chemical force microscopy and monitoring of nanoparticle capture.
Engineered peptides capable of binding to silica have been used to provide contrast in chemical force microscopy and tested for their capacity to selectively capture silica nanoparticles (NPs). Gold coated atomic force microscopy (AFM) microcantilevers with integrated tips and colloidal probes were functionalized with engineered peptides through a thiol group of a terminal cysteine which was linked via a glycine trimer to a 12-mer binding sequence. The functionalized probes demonstrated a significantly increased binding force on silicon oxide areas of a gold-patterned silicon wafer, whereas plain gold probes, and those functionalized with a random permutation of the silica binding peptide motif or an all-histidine sequence displayed similar adhesion forces to gold and silicon oxide. As the functionalized probes also allowed contact mode imaging subsequently to the adhesion mapping, also the associated friction contrast was measured and found to be similar to the adhesion contrast. Furthermore, the adsorption of silica NPs onto planar gold surfaces functionalized in the same manner was observed to be selective. Notably, the surface coverage with silica NPs was found to decrease with increasing pH, implying the importance of electrostatic interactions between the peptide and the NPs. Finally, the adsorption of silica NPs was monitored via the decrease in fundamental resonance frequency of an AFM microcantilever functionalized with silica binding peptides.


Influence of the autonomic nervous system on calcium homeostasis in the rat.
The local surgical manipulation of sympathetic and parasympathetic nerves innervating the thyroid-parathyroid territory was employed to search for the existence of a peripheral neuroendocrine link controlling parathyroid hormone (PTH) and calcitonin (CT) release. From 8 to 24 h after superior cervical ganglionectomy (SCGx), at the time of wallerian degeneration of thyroid-parathyroid sympathetic nerve terminals, an alpha-adrenergic inhibition, together with a minor beta-adrenergic stimulation, of hypercalcemia-induced CT release, and an alpha-adrenoceptor inhibition of hypocalcemia-induced PTH release were found. In chronically SCGx rats PTH response to EDTA was slower, and after CaCl2 injection, serum calcium attained higher levels in face of normal CT levels. SCGx blocked the PTH increase found in sham-operated rats stressed by a subcutaneous injection of turpentine oil, but did not affect the greater response to EDTA. The higher hypocalcemia seen after turpentine oil was no longer observed in SCGx rats. The effects of turpentine oil stress on calcium and CT responses to a bolus injection of CaCl2 persisted in rats subjected to SCGx 14 days earlier. Interruption of thyroid-parathyroid parasympathetic input conveyed by the thyroid nerves (TN) and the inferior laryngeal nerves (ILN) caused a fall in total serum calcium, an increase of PTH levels and a decrease of CT levels, when measured 10 days after surgery. Greater responses of serum CT and PTH were detected in TN-sectioned, and in TN- or ILN-sectioned rats, respectively. Physiological concentrations of CT decreased, and those of PTH increased, in vitro cholinergic activity in rat SCG, measured as specific choline uptake, and acetylcholine synthesis and release. The results indicate that cervical autonomic nerves constitute a pathway through which the brain modulates calcium homeostasis.


Autophagy balances inflammation in innate immunity.
Macroautophagy/autophagy is a homeostatic process with multiple effects on immunity. One of the pivotal contributions of autophagy in immunity is the cell autonomous control of inflammation. This property leads to systemic consequences and thereby influences the development of innate and adaptive immunity, which promotes or suppresses pathology in various disease contexts. In this review we focus on the intersections between autophagy and inflammasome activation, autophagy and interferons, and autophagy and inflammation in association with infection.


Statistical analysis of crossover interference using the chi-square model.
The chi-square model (also known as the gamma model with integer shape parameter) for the occurrence of crossovers along a chromosome was first proposed in the 1940's as a description of interference that was mathematically tractable but without biological basis. Recently, the chi-square model has been reintroduced into the literature from a biological perspective. It arises as a result of certain hypothesized constraints on the resolution of randomly distributed crossover intermediates. In this paper under the assumption of no chromatid interference, the probability for any single spore or tetrad joint recombination pattern is derived under the chi-square model. The method of maximum likelihood is then used to estimate the chi-square parameter m and genetic distances among marker loci. We discuss how to interpret the goodness-of-fit statistics appropriately when there are some recombination classes that have only a small number of observations. Finally, comparisons are made between the chi-square model and some other tractable models in the literature.


Risk Adjustment in ALPPS Is Associated With a Dramatic Decrease in Early Mortality and Morbidity.
To longitudinally assess whether risk adjustment in Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) occurred over time and is associated with postoperative outcome. ALPPS is a novel 2-stage hepatectomy enabling resection of extensive hepatic tumors. ALPPS has been criticized for its high mortality, which is reported beyond accepted standards in liver surgery. Therefore, adjustments in patient selection and technique have been performed but have not yet been studied over time in relation to outcome. ALPPS centers of the International ALPPS Registry having performed ≥10 cases over a period of ≥3 years were assessed for 90-day mortality and major interstage complications (≥3b) of the longitudinal study period from 2009 to 2015. The predicted prestage 1 and 2 mortality risks were calculated for each patient. In addition, questionnaires were sent to all centers exploring center-specific risk adjustment strategies. Among 437 patients from 16 centers, a shift in indications toward colorectal liver metastases from 53% to 77% and a reverse trend in biliary tumors from 24% to 9% were observed. Over time, 90-day mortality decreased from initially 17% to 4% in 2015 (P = 0.002). Similarly, major interstage complications decreased from 10% to 3% (P = 0.011). The reduction of 90-day mortality was independently associated with a risk adjustment in patient selection (P < 0.001; OR: 1.62; 95% CI: 1.36-1.93) and using less invasive techniques in stage-1 surgery (P = 0.019; OR: 0.39; 95% CI: 0.18-0.86). A survey indicated risk adjustment of patient selection in all centers and ALPPS technique in the majority (80%) of centers. Risk adjustment of patient selection and technique in ALPPS resulted in a continuous drop of early mortality and major postoperative morbidity, which has meanwhile reached standard outcome measures accepted for major liver surgery.


Perinatal exposure to low levels of the environmental antiandrogen vinclozolin alters sex-differentiated social play and sexual behaviors in the rat.
In this study we examined the effects of exposure to the antiandrogenic fungicide vinclozolin (Vz) on the development of two sex-differentiated behaviors that are organized by the perinatal actions of androgens. Pregnant Long-Evans rats were administered a daily oral dose of 0, 1.5, 3, 6, or 12 mg/kg Vz from the 14th day of gestation through postnatal day (PND)3. The social play behavior of juvenile offspring was examined on PND22 and again on PND34 during play sessions with a same-sex littermate. After they reached adulthood, the male offspring were examined with the ex copula penile reflex procedure to assess erectile function. Vz did not produce any gross maternal or neonatal toxicity, nor did it reduce the anogenital distance in male pups. We observed no effects of Vz on play behavior on PND22. However, the 12-mg/kg Vz dose significantly increased play behavior in the male offspring on PND34 compared with controls. The most dramatic increases were seen with the nape contact and pounce behavior components of play. The Vz effect was more pronounced in male than in female offspring. As adults, male offspring showed a significant reduction of erections at all dose levels during the ex copula penile reflex tests. The 12-mg/kg dose was also associated with an increase in seminal emissions. These effects demonstrate that perinatal Vz disrupts the development of androgen-mediated behavioral functions at exposure levels that do not produce obvious structural changes or weight reductions in androgen-sensitive reproductive organs.


LC-MS/MS for Identifying Patients with CYP24A1 Mutations.
Patients have been described with loss-of-function CYP24A1 (cytochrome P450, family 24, subfamily A, polypeptide 1) mutations that cause a high ratio of 25-hydroxyvitamin D to 24,25-dihydroxyvitamin D [25(OH)D/24,25(OH)2D], increased serum 1,25-dihydroxyvitamin D, and resulting hypercalcemia, hypercalciuria and nephrolithiasis. A 25(OH)D/24,25(OH)2D ratio that can identify patients who are candidates for confirmatory CYP24A1 genetic testing would be valuable. We validated an LC-MS/MS assay for 24,25(OH)2D (D3 and D2) and determined a 25(OH)D/24,25(OH)2D cutoff to identify candidates for confirmatory genetic testing. After addition of isotope-labeled internal standard, serum samples were extracted by solid-phase extraction, derivatized with 4-phenyl-1,2,4,-triazoline-3,5-dione, and quantified by LC-MS/MS. We measured 25(OH)D/24,25(OH)2D in 91 healthy patients and 34 patients with clinically suspected CYP24A1-mediated hypercalcemia. The limits of detection and quantification were 0.03 (0.2) and 0.1 (0.24) nmol/L, respectively, for 24,25(OH)2D3, and 0.1 (0.23) and 0.5 (1.16) nmol/L for 24,25(OH)2D2. Intra- and interassay imprecision was 4%-15% across the analytical measurement range of 0.1-25 ng/mL (0.2-60 nmol/L). No interference was observed with 25(OH)D and 1,25(OH)2D. 25(OH)D/24,25(OH)2D of 7-35 was observed in healthy patients, whereas in 2 patients with CYP24A1 mutations, 25(OH)D/24,25(OH)2D was significantly increased (99-467; P < 0.001). A 25(OH)D/24,25(OH)2D ratio ≥99 identified patients who were candidates for CYP24A1 genetic testing. Increased 25(OH)D/24,25(OH)2D supports the diagnosis of reduced CYP24A1 activity due to mutations in CYP24A1. Measurement of 25(OH)D/24,25(OH)2D should be considered a part of the clinical workup in patients with hypercalcemia of otherwise unknown etiology.


Transcriptome of the Australian Mollusc Dicathais orbita Provides Insights into the Biosynthesis of Indoles and Choline Esters.
Dicathais orbita is a mollusc of the Muricidae family and is well known for the production of the expensive dye Tyrian purple and its brominated precursors that have anticancer properties, in addition to choline esters with muscle-relaxing properties. However, the biosynthetic pathways that produce these secondary metabolites in D. orbita are not known. Illumina HiSeq 2000 transcriptome sequencing of hypobranchial glands, prostate glands, albumen glands, capsule glands, and mantle and foot tissues of D. orbita generated over 201 million high quality reads that were de novo assembled into 219,437 contigs. Annotation with reference to the Nr, Swiss-Prot and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases identified candidate-coding regions in 76,152 of these contigs, with transcripts for many enzymes in various metabolic pathways associated with secondary metabolite biosynthesis represented. This study revealed that D. orbita expresses a number of genes associated with indole, sulfur and histidine metabolism pathways that are relevant to Tyrian purple precursor biosynthesis, and many of which were not found in the fully annotated genomes of three other molluscs in the KEGG database. However, there were no matches to known bromoperoxidase enzymes within the D. orbita transcripts. These transcriptome data provide a significant molecular resource for gastropod research in general and Tyrian purple producing Muricidae in particular.


Structure prediction of ordered and disordered multiple octahedral cation perovskites using SPuDS.
The software package SPuDS has previously been shown to accurately predict crystal structures of AMX(3) and A(1 - x)A'(x)MX(3) perovskites that have undergone octahedral tilting distortions. This paper describes the extension of this technique and its accuracy for A(2)MM'X(6) ordered double perovskites with the aristotype Fm\overline 3m cubic structure, as well as those that have undergone octahedral tilting distortions. A survey of the literature shows that roughly 70% of all ordered double perovskites undergo octahedral tilting distortions. Of the 11 distinct types of octahedral tilting that can occur in ordered perovskites, five tilt systems account for approximately 97% of the reported structures. SPuDS can calculate structures for the five dominant tilt systems, Fm\overline 3m (a(0)a(0)a(0)), I4/m (a(0)a(0)c(-)), R\overline 3 (a(-)a(-)a(-)), I2/m (a(0)b(-)b(-)) and P2(1)/n (a(-)a(-)b(+)), as well as two additional tilt systems, Pn\overline 3 (a(+)a(+)a(+)) and P4/mnc (a(0)a(0)c(+)). Comparison with reported crystal structures shows that SPuDS is quite accurate at predicting distortions driven by octahedral tilting. The favored modes of octahedral tilting in ordered double perovskites are compared and contrasted with those in AMX(3) perovskites. Unit-cell pseudosymmetry in Sr- and Ca-containing double perovskites is also examined. Experimentally, Sr(2)MM'O(6) compounds show a much stronger tendency toward pseudosymmetry than do Ca(2)MM'O(6) compounds with similar tolerance factors.


The PAX8 cistrome in epithelial ovarian cancer.
PAX8 is a lineage-restricted transcription factor that is expressed in epithelial ovarian cancer (EOC) precursor tissues, and in the major EOC histotypes. Frequent overexpression of PAX8 in primary EOCs suggests this factor functions as an oncogene during tumorigenesis, however, the biological role of PAX8 in EOC development is poorly understood. We found that stable knockdown of PAX8 in EOC models significantly reduced cell proliferation and anchorage dependent growth in vitro, and attenuated tumorigenicity in vivo. Chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq) and transcriptional profiling were used to create genome-wide maps of PAX8 binding and putative target genes. PAX8 binding sites were significantly enriched in promoter regions (p < 0.05) and superenhancers (p < 0.05). MEME-ChIP analysis revealed that PAX8 binding sites overlapping superenhancers or enhancers, but not promoters, were enriched for JUND/B and ARNT/AHR motifs. Integrating PAX8 ChIP-seq and gene expression data identified PAX8 target genes through their associations within shared topological association domains. Across two EOC models we identified 62 direct regulatory targets based on PAX8 binding in promoters and 1,330 putative enhancer regulatory targets. SEPW1, which is involved in oxidation-reduction, was identified as a PAX8 target gene in both cell line models. While the PAX8 cistrome exhibits a high degree of cell-type specificity, analyses of PAX8 target genes and putative cofactors identified common molecular targets and partners as candidate therapeutic targets for EOC.


Skeletal muscle interleukin-6 regulates metabolic factors in iWAT during HFD and exercise training.
To investigate the role of skeletal muscle (SkM) interleukin (IL)-6 in the regulation of adipose tissue metabolism. Muscle-specific IL-6 knockout (IL-6 MKO) and IL-6(loxP/loxP) (Floxed) mice were subjected to standard rodent diet (Chow), high-fat diet (HFD), or HFD in combination with exercise training (HFD ExTr) for 16 weeks. Total fat mass increased (P < 0.05) in both genotypes with HFD. However, HFD IL-6 MKO mice had lower (P < 0.05) inguinal adipose tissue (iWAT) mass than HFD Floxed mice. Accordingly, iWAT glucose transporter 4 (GLUT4) protein content, 5'AMP activated protein kinase (AMPK)(Thr172) phosphorylation, and fatty acid synthase (FAS) mRNA content were lower (P < 0.05) in IL-6 MKO than Floxed mice on Chow. In addition, iWAT AMPK(Thr172) and hormone-sensitive lipase (HSL)(Ser565) phosphorylation as well as perilipin protein content was higher (P < 0.05) in HFD IL-6 MKO than HFD Floxed mice, and pyruvate dehydrogenase E1α (PDH-E1α) protein content was higher (P < 0.05) in HFD ExTr IL-6 MKO than HFD ExTr Floxed mice. These findings indicate that SkM IL-6 affects iWAT mass through regulation of glucose uptake capacity as well as lipogenic and lipolytic factors.


Activation of microcomponents with light for micro-electro-mechanical systems and micro-optical-electro-mechanical systems applications.
We examine the light-activation properties of micrometer-sized gear structures fabricated with polysilicon surface micromachining techniques. The gears are held in place on a substrate through a capped anchor post and are free to rotate about the post. The light-activation technique is modeled on photon radiation pressure, and the equation of motion of the gear is solved for this activation technique. Experimental measurements of torque and damping are found to be consistent with expected results for micrometer-scale devices. Design optimization for optically actuated microstructures is discussed.


Modeling of stimulation-secretion coupling in a chromaffin cell.
We constructed a chromaffin cell model for analysis of stimulation-secretion coupling in computer simulation studies. The model includes mechanisms involved in the excitatory synapse, voltage-dependent Na(+), K(+) and Ca(2+) channels, Ca(2+)-activated K(+) channels (SK type), buffered Ca(2+) diffusion, Ca(2+) extrusion, fluorescent Ca(2+) indicators and Ca(2+)-triggered exocytosis. Calculations of the modeled mechanisms were carried out using the NEURON simulation environment (Hines and Carnevale, Neural Computation 9:1179-1209, 1997). A set of parameter values was determined so as to fit basic experimental results reported in the literature. The model was also applied to simulate our experimental results obtained from chromaffin cells in the perfused rat adrenal medulla. Observed profiles of Ca(2+)responses induced by electrically stimulating the splanchnic nerve with various frequencies (1-50 Hz) were adequately simulated with minor readjustments of parameter values for Ca(2+)influx and extrusion. Secretory responses measured at the same time as the Ca(2+)responses were also simulated with consideration of a time constant to detect catecholamines in the experiment. Similarly, model simulations reproduced both Ca(2+)responses and secretory responses evoked by elevations of the extracellular K(+) concentration for different periods. The results suggest that the presented model provides a useful tool for analyzing and predicting quantitative relations in various events occurring in stimulation-secretion coupling in chromaffin cells.


Pharmacological, antioxidant, genotoxic studies and modulation of rat splenocyte functions by Cyperus rotundus extracts.
Cyperus rotundus Linn. (Cyperaceae) is a Tunisian medicinal plant used in folkloric (traditional) medicine to treat stomach disorders and inflammatory diseases. The present study explored the analgesic, anti-inflammatory and genotoxic activities of extracts from the aerial parts of C. rotundus. The antioxidant capacity and the modulation of splenocyte functions by these extracts were also investigated in mice. The phytochemical analysis was carried out using standard methods. Aqueous, ethyl acetate, methanol and TOF-enriched extracts (300, 150, and 50 μg/ml) were evaluated for their analgesic and anti-inflammatory activities. 4, 2, and 1 mg/ml of each extract were tested to investigate their effect on lipid peroxidation. The genotoxic study was monitored by measuring the structural chromosome aberrations of mice treated with 300 mg/kg of extract. The proliferation of lymphocytes in the absence and presence of mitogens was assessed at a concentration range 1-1000 μg/ml. The tested extracts were able to decrease the mouse ear oedema induced by xylene. Furthermore, it was shown that the same extracts reduced the number of abdominal contractions caused by acetic acid in mice, revealing the peripheral analgesic activity of these extracts. It is worth noting that mice treated with doses up to 300 mg/kg b.w. of Cyperus rotundus extracts did not exhibit any toxicity. The tested extracts significantly enhance lymphocyte proliferation at 1 mg/ml. It appears that C. rotundus extracts contain potent components such as flavonoids that may potentially be useful for modulating the immune cell functions, provoking analgesic, anti-inflammatory and antioxidant effects.