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The recent discoveries of the involvement of SK3 channel in some cell motility mechanisms occurring in cancer disease have opened up the way to the synthesis of inhibitors that could reduce metastasis formation. On the basis of our recent previous works showing that both lactose-glycero-ether lipid (Ohmline) and some phosphate analogues (GPGEL) were efficient compounds to modulate SK3 channel activity, the present study, which found its inspiration in the structure of the natural glycolipid DiGalactosylDiacylGlycerol (DGDG), reports the incorporation of a digalactosyl moiety (á-galactopyranosyl-(1?6)-â-galactopyranosyl-) as the polar head of a glycero ether lipid. For the construction of the digalactosyl fragment, two synthetic approaches were compared. The standard strategy which is based on the use of the benzyl protecting group to produce 1?6 disaccharide unit, was compared with a second method that made use of the trimethylsilyl moiety as a protecting group. This second strategy, which is applied for the first time to the synthesis of (1?6)-disaccharide unit, presents a net advantage in terms of efficacy (better global yield) and cost. Finally, compound 16, which is characterized by a (1?6) DiGalactosyl unit (DG) as the polar head of the amphiphilic structure, was tested as a modulator of the SK3 channel activity. Patch-clamp experiments have shown that compound 16 reduced SK3 currents (-28.2 ± 2.0% at 5 ìM) and cell migration assays performed at 300 nM have shown a reduction of cell migration (SK3 + HEK293T) by 19.6 ± 2.7%.

BACKGROUND: Accurate quantification of DNA using quantitative real-time PCR at low levels is increasingly important for clinical, environmental and forensic applications. At low concentration levels (here referring to under 100 target copies) DNA quantification is sensitive to losses during preparation, and suffers from appreciable valid non-detection rates for sampling reasons. This paper reports studies on a real-time quantitative PCR assay targeting a region of the human SRY gene over a concentration range of 0.5 to 1000 target copies. The effects of different sample preparation and calibration methods on quantitative accuracy were investigated.RESULTS: At very low target concentrations of 0.5-10 genome equivalents (g.e.) eliminating any replicates within each DNA standard concentration with no measurable signal (non-detects) compromised calibration. Improved calibration could be achieved by eliminating all calibration replicates for any calibration standard concentration with non-detects ('elimination by sample'). Test samples also showed positive bias if non-detects were removed prior to averaging; less biased results were obtained by converting to concentration, including non-detects as zero concentration, and averaging all values. Tube plastic proved to have a strongly significant effect on DNA quantitation at low levels (p = 1.8 x 10(-4)). At low concentrations (under 10 g.e.), results for assays prepared in standard plastic were reduced by about 50% compared to the low-retention plastic. Preparation solution (carrier DNA or stabiliser) was not found to have a significant effect in this study.Detection probabilities were calculated using logistic regression. Logistic regression over large concentration ranges proved sensitive to non-detected replicate reactions due to amplification failure at high concentrations; the effect could be reduced by regression against log (concentration) or, better, by eliminating invalid responses.CONCLUSION: Use of low-retention plastic tubes is advised for quantification of DNA solutions at levels below 100 g.e. For low-level calibration using linear least squares, it is better to eliminate the entire replicate group for any standard that shows non-detects reasonably attributable to sampling effects than to either eliminate non-detects or to assign arbitrary high Ct values. In calculating concentrations for low-level test samples with non-detects, concentrations should be calculated for each replicate, zero concentration assigned to non-detects, and all resulting concentration values averaged. Logistic regression is a useful method of estimating detection probability at low DNA concentrations.

INTRODUCTION: In conjunction with adjuvant chemotherapy, radical resections are the only treatment modality, which significantly prolongs survival in pancreatic cancer (CaP) patients. The author aims to define current standards of radical pancreatic resections in CaP patients and to assess benefits of the surgical procedure based on literature data, as well as to evaluate current options for objective assessment of the quality of life in these patients.METHODS: Employing Pubmed and Ebscohost databases, the author compares radical pancreatic resections depending on the tumor location, performed in conjunction with standard lymphadenectomy in patients with CaP, with the extended version of the resection procedure. The radical procedure is then put into relationship with potential prolongation of survival times, with early mortality incidence rates and with rates of perioperative complications. Based on literature data, the author evaluates current options for objective assessment of the quality of life in these patients.OUTCOMES: Radical resection with lymphadenectomy in N1-2 diseases followed by adjuvant chemotherapy is currently considered a standard treatment procedure, the only one which significantly prolongs survival in patients with CaP, with the disease stage T1-3, N0-1M0. Angioinvasion into the portomesenteric segment is not a contraindication for the resection procedure. In cases where R0 resection is achieved, the outcomes are similar to those in subjects without vascular segment resections. Views on arterial resections--a. hepatica, truncus coeliacus, a. mesenterica sup. are not uniform and, to date, resections of tumor-infiltrated arteries have not been shown to result in life prolongation. Angioinvasion into arteries is considered a sign of the disease stage and the need for extended procedures is associated with increased complication rates. The quality of life following resection procedures is not commonly assessed and its improvement is usually expected, rather than objectively assessed. At the present time, no questionnaire on the quality of life, which would specifically address CaP patients, is being used on a routine basis.DISCUSSION AND CONCLUSIONS: Radical resection R0 and administration of adjuvant chemotherapy is the only current treatment modality in patients with ductal pancreatic adenocarcinoma, which results in significant life prolongation with 7-25% five-year survival rates (median of 15-18.5 months). The tumor's biological characteristics and the fact that the real disease staging is difficult to establish, are the reasons for early relapses after so called R0 resections. Resections of the portomesenteric segment in cases with adherence or penetration of the CaP into the vascular wall is indicated as a part of the radical PDE and do not result in shorter survival times. Extended radical procedures with arterial resections (a. hepatica, truncus coeliacus, a. mesenterica sup.) do not result in prolonged survival times even in cases where the procedures were assessed as R0 procedures, and cannot be thought of as standard procedures. By using invasive resection procedures, the number of resecable patients would increase, however, the prize of this treatment decision may need to paid off by higher complication rates, compared to those in standard procedures, as well as by lower postoperative quality of life of the patients, while achieving comparable survival time outcomes.

Twelve arrhythmias that occurred during and following surgery for acquired heart disease were studied in a group of 52 randomly selected patients. In cases of aortic and mitral valve replacement, Bj?rk-Shiley prosthetic valves were used, and procain-magnesium-aspartate solution was applied for cardiac arrest. Registration and analysis of rhythm disturbances were carried out at 10 different time intervals, spaced narrowly in the intra-and early postoperative phases, then more widely up to the 96th hour of observation. Arrhythmic incidence and occurrence per case and phase are given, thereby allowing an account of transient arrhythmias as well. The peak occurrence of serious rhythm disturbance was found, intraoperatively, at about the time of aortic cross-clamping and release (ischaemic effect) with the exception of the mitral-commissurotomy group. The 1st-6th postbypass hours showed an arrhythmia trough, followed by a 2nd peak (9th-96th postoperative hour) made up of less serious arrhythmias (electrolyte- and acid-base disturbances; digitalis effects). All 12 arrhythmia types, as well as the five separately considered ventricular arrhythmias of the "first arrhythmia peak", did not reveal any significant relation to: patients' age, N.Y.H.A. criteria, pre-operative PAm or the duration of anaesthesia, surgery and bypass time.

The involvement of phospholipase D (PLD) in phosphatidylcholine hydrolysis by epidermal (EGF), insulin-like (IGF-I), and basic fibroblast (bFGF) growth factors was investigated in rat pancreatic acini. Acini were prelabeled with [3H]myristic acid which is mostly incorporated into phosphatidylcholine. EGF, IGF-I, and bFGF caused significant and dose-dependent increases in [3H]phosphatidic acid (PA) accumulation in the presence of propranolol, a phosphatidic acid phosphohydrolase inhibitor. The effects of EGF and IGF-I were significant after 5, 15, and 30 min of stimulation, whereas that of bFGF was evident only at 30 min. PA production in response to all three factors was dose dependent with maximal responses to EGF at 25 nM, to IGF-I at 16.5 nM, and to bFGF at 50 pM. Preincubation of acini with staurosporine, a protein kinase C and tyrosine kinase inhibitor, totally inhibited PA production by the three factors. Similarly, acini preincubation with genistein, a specific tyrosine kinase inhibitor, also neutralized the influence of the three factors on PA accumulation. In the presence of 1% ethanol, EGF, IGF-I, and bFGF caused significant phosphatidylethanol production after 20 min of incubation, thus confirming the involvement of PLD in PA production. These data present for the first time the description of a new signaling pathway through which EGF, IGF-I, and bFGF may operate to induce some of their specific effects on the pancreas in association with these growth factor receptors' tyrosine kinase activity.

This is the second of two articles providing an introduction to the critical appraisal of research reports. In this paper guidelines for evaluating the quality of evidence and the validity of conclusions are suggested, and a check-list of evaluation points is presented.

Antibody-drug conjugates (ADCs) are biochemotherapeutics consisting of a cytotoxic chemical drug linked covalently to a monoclonal antibody. Two main classes of ADCs, namely cysteine and lysine conjugates, are currently available on the market or involved in clinical trials. The complex structure and heterogeneity of ADCs makes their biophysical characterization challenging. For cysteine conjugates, hydrophobic interaction chromatography is the gold standard technique for studying drug distribution, the naked antibody content, and the average drug to antibody ratio (DAR). For lysine ADC conjugates on the other hand, which are not amenable to hydrophobic interaction chromatography because of their higher heterogeneity, denaturing mass spectrometry (MS) and UV/Vis spectroscopy are the most powerful approaches. We report here the use of native MS and ion mobility (IM-MS) for the characterization of trastuzumab emtansine (T-DM1, Kadcyla(®)). This lysine conjugate is currently being considered for the treatment of human epidermal growth factor receptor 2 (HER2)-positive breast cancer, and combines the anti-HER2 antibody trastuzumab (Herceptin(®)), with the cytotoxic microtubule-inhibiting maytansine derivative, DM1. We show that native MS combined with high-resolution measurements and/or charge reduction is beneficial in terms of the accurate values it provides of the average DAR and the drug load profiles. The use of spectral deconvolution is discussed in detail. We report furthermore the use of native IM-MS to directly determine DAR distribution profiles and average DAR values, as well as a molecular modeling investigation of positional isomers in T-DM1.

The bifunctional, microbial enzyme DmpFG is comprised of two subunits: the aldolase, DmpG, and the dehydrogenase, DmpF. DmpFG is of interest due to its ability to channel substrates between the two spatially distinct active sites. While the aldolase is well studied, significantly less is known about the dehydrogenase. Steady-state kinetic measurements of the reverse reaction of DmpF confirmed that the dehydrogenase uses a ping-pong mechanism, with substrate inhibition by acetyl CoA indicating that NAD(+)/NADH and CoA/acetyl CoA bind to the same site in DmpF. The Km of DmpF for exogenous acetaldehyde as a substrate was 23.7 mM, demonstrating the necessity for the channel to deliver acetaldehyde directly from the aldolase to the dehydrogenase active site. A channeling assay on the bifunctional enzyme gave an efficiency of 93% indicating that less than 10% of the toxic acetaldehyde leaks out of the channel into the bulk media, prior to reaching the dehydrogenase active site. The thermodynamic activation parameters of the reactions catalyzed by the aldolase, the dehydrogenase and the DmpFG complex were determined. The Gibb's free energy of activation for the dehydrogenase reaction was lower than that obtained for the full DmpFG reaction, in agreement with the high kcat obtained for the dehydrogenase reaction in isolation. Furthermore, although both the DmpF and DmpG reactions occur with small, favorable entropies of activation, the full DmpFG reaction occurs with a negative entropy of activation. This supports the concept of allosteric structural communication between the two enzymes to coordinate their activities.

The presence of intestinal helminths in the alimentary canal of brown trout, Salmo trutta L., can alter the number of cells that synthesize modulatory peptides. A total of 167 brown trout were collected from tributaries of the River Brenta (northern Italy), of which 119 (71.3%) specimens were infected with enteric helminths, 28 with the acanthocephalan Pomphorhynchus laevis M?ller, 1776 with intensity of infection ranging from 1 to 162 (18.57 +/- 30.79) worms per host and 67 fish with the cestode Cyathocephalus truncatus Pallas, 1781. Intensity of infection with C. truncatus ranged from 1 to 85 (6.87 +/- 12.59) per fish. In 24 fish there were concurrent infections of both species of helminths. The caecal and middle regions of the intestine were the most heavily parasitized. Immunohistochemical tests showed a decrease in endocrine cells (ECs) of the diffuse endocrine system (DES) positive to gastrin, cholecystokinin-8, bombesin and secretin antisera in the intestine of the infected trout. The number of ECs immunoreactive to anti-glucagon serum did not show differences in the digestive tract of uninfected brown trout and in conspecifics parasitized with P. laevis. The density of cells containing glucagon-like material was low in the fish parasitized with C. truncatus. The results suggest that endoparasitic helminths induce alterations in the DES of infected S. trutta.

BACKGROUND: Dietitians and other health care professionals must be able to translate findings from clinical trials into best treatment practices, a skill termed "knowledge translation". This skill requires knowledge of treatment guidelines as well as the science underpinning treatment recommendations. Unsatisfactory knowledge translation of medical nutrition therapy (MNT) has been documented.METHODS: Individuals registered to attend a leading national nutrition conference were asked to participate in an online cross-sectional survey. Participants were asked to provide demographic and professional information, answer questions on nutrition knowledge and to choose a clinical action plan in response to dietitian-designed case vignettes describing research outcomes. Responses were compared by profession and participation in research activities.RESULTS: Of 3000 registered conference attendees, 299 individuals replied: 79.0% dietitians, 93.3% female, with a mean household income matching the 5th decile of income, 60.7% indicated a single employment setting, 20.7% reported participating in research. Almost 74% of respondents indicated that they would make clinical recommendations based on findings of an in vitro study. In one vignette, a patient with a disease previously not encountered by the respondent required a clinical treatment plan. Only 53% of participants chose to seek formal nutrition guidelines. Fewer than 15% of participants could identify the pathway for fat during weight loss. Differences in knowledge translation skills by research participation were not detected.CONCLUSIONS: Our findings reveal a deficit in knowledge translation proficiency in a convenience sample of dietitians and other health professionals, highlighting the need to develop these skills.

BACKGROUND AND STUDY AIM: Intrapapillary capillary loops (IPCLs) show distinct pattern changes corresponding to tumor progression and depth of invasion, important for in vivo characterization of superficial squamous cell carcinoma (SCC). We examined the relation between invasion depth and histopathologic IPCL diameter.PATIENTS AND METHODS: Prospectively, before lesion resection, magnification endoscopy and narrow band imaging were used to identify IPCL patterns of type V1 (corresponding to tumors limited to the mucosa; 10 patients) and type Vn (submucosally invading tumors; 10 patients). Post-resection, IPCL samples (type I [normal mucosa], n = 103; V1, n = 113; Vn, n = 100) were stained with hematoxylin & eosin, CD34, and desmin, and vessel diameter measured using light microscopy.RESULTS: Mean (standard deviation [SD]) histopathologic calibers of IPCLs of types I, V1, and Vn were significantly different, being 7.7 (2.8) µm, 21.9 (7.4) µm, and 65.2 (22.9) µm; type 1 vs. V1, P < 0.001; V1 vs. Vn, P < 0.001.CONCLUSIONS: Magnification endoscopy observation of IPCLs allows in vivo discrimination between intramucosal and submucosally invasive cancer.

Background:: Hidradenitis is a chronic inflammatory disease of the hair follicles. A treatment is necessary due to chronicity and psychological changes that patient present.Objective:: To investigate epidemiological aspects and elaborate a risk group profile, promote early diagnosis and contribute to the knowledge about the disease.Methods:: This cross-sectional descriptive study with retrospective analysis of medical records of 194 patients diagnosed with hidrosadenitis in a dermatological reference center in the city of Bauru (SP) between 2005 and 2015.Results:: Females accounted for 74% of cases. The age at diagnosis ranged from 10 to 67 years and the majority was within the 3rd and 4th decade of life. It occurred Association with diabetes mellitus in 33%, obesity in 55% and smoking in 61% was observed. Mean time between the onset of the disease and diagnosis was nine years. Hurley stage II was the most common at diagnosis. The therapeutic option mostly used in Hurley I and II was systemic antibiotics and in Hurley III was surgery.Study limitations:: the main limitation of this study is its retrospective design, which does not allow the true clinical confirmation of the disease by investigators.Conclusion:: we outlined the following profile: women, caucasian, between 3rd and 4th decade of life, associated with obesity, smoking, late diagnosis and multiple potential therapeutic modalities. We highlight the importance of studies like this in order to identify risk groups and encourage early diagnosis.

AIM: To evaluate the feasibility of the revised "Clinical Guideline for HIV and TB" in the Great Tehran Prison during October 2013 to June 2014.METHODS: The guideline includes all aspects of HIV/TB diagnosis based on active case finding (ACF), treatment and care services. Before the implementation, a focus group discussion was conducted, and attended by experts on prison health. The objective was to identify defects and limitations of the guideline. After the discussion, the guideline was revised. The Great Tehran Prison contains three separate units; all prisoners are taken first to "reception and identification unit (quarantine)" and then send to two housing units according to their legal status. An HIV ACF strategy was employed in the quarantine, and two units through a voluntary provider-initiated HIV testing. Three staff of the triangular clinic trained the prisoners about common routes of HIV transmission and the symptoms of TB in the units. In the quarantine, all prisoners were examined for all HIV-risk factors, HIV testing and symptoms of TB. In unit one, healthcare staff continued the ACF process, while in unit two, the peers of prisoners were assigned as the healthcare communicators to proceed with the strategy. At this caring process, when the test result was positive, then the process of care, treatment and follow ups was initiated. Moreover, the use of directly observed therapy (DOT) for antiretroviral therapy (ART) and TB was applied to the sick prisoners. There was also a follow-up caring for released prisoner to refer them to care and treatment services outside the prison.RESULTS: The guideline was implemented in the prison successfully.CONCLUSION: Regarding feasibility of the guideline, the investigators of this study suggest that the guideline should be implemented in other prisons across the country.

Triple-negative breast tumors are very aggressive and contain relatively high proportion of cancer stem cells, and are resistant to chemotherapeutic drugs including cisplatin. To overcome these limitations, we combined eugenol, a natural polyphenolic molecule, with cisplatin to normalize cisplatin mediated toxicity and potential drug resistance. Interestingly, the combination treatment provided significantly greater cytotoxic and pro-apoptotic effects as compared to treatment with eugenol or cisplatin alone on several triple-negative breast cancer cells both in vitro and in vivo. Furthermore, adding eugenol to cisplatin potentiated the inhibition of breast cancer stem cells by inhibiting ALDH enzyme activity and ALDH-positive tumor initiating cells. We provide also clear evidence that eugenol potentiates cisplatin inhibition of the NF-êB signaling pathway. Indeed, the binding of NF-êB to its cognate binding sites present in the promoters of IL-6 and IL-8 was dramatically reduced, which led to potent down-regulation of the IL-6 and IL-8 cytokines upon combination treatment relative to the single agents. Similar effects were observed on proliferation, inhibition of epithelial-to-mesenchymal transition and stemness markers in tumor xenografts. These results provide strong preclinical justification for combining cisplatin with eugenol as therapeutic approach for triple-negative breast cancers through targeting the resistant ALDH-positive cells and inhibiting the NF-êB pathway.

OBJECTIVE: This study analyzed the outcomes of a series of consecutive patients diagnosed with a retroperitoneal soft tissue sarcoma (RSTS) with vascular involvement and who underwent a multidisciplinary operation.METHODS: Between 2000 and 2013, 126 patients were referred for oncovascular surgery in our institution. Among these, 31 consecutive patients underwent operations for RSTS with vascular involvement. A vascular/oncologic team determined the surgical strategy preoperatively.RESULTS: Median follow-up was 34.4 months (interquartile range, 48.1 months). Twenty patients (65%) were referred for primary RSTS and 11 (35%) for local recurrence. The most common histologic diagnosis was liposarcoma (54.8%), mainly high-grade and intermediate-grade RSTS. Prosthetic grafts were usually used for vascular reconstruction. Median hospital stay was 17 days (interquartile range, 14.5; range, 7-190 days). The grade 3 and 4 morbidity rate was 19.3%. Each resection was macroscopically complete (R0-R1). Median progression-free survival was 10 months, and median overall survival was not reached. Overall survival rates were 77.4% at 1 year and 61.3% at 3 years.CONCLUSIONS: Vascular resection and reconstruction are safe and feasible in case of RSTS. The morbidity rate was acceptable, and there were no perioperative deaths. Despite recurrence rates that remain high, oncovascular resection enhances resection margins and allows encouraging survival results for patients often considered as nonresectable.

To clarify the role of p53 in the development of Epstein-Barr virus (EBV)-associated gastric carcinoma, we examined DNA-ploidy pattern, the immunoreactivity of p53, p21(WAF1) and Ki-67 and the incidence of mitosis and apoptosis in EBV-positive and -negative gastric carcinomas of similar histology: a lace pattern and/or gastric carcinoma with lymphoid stroma. There was no significant difference in DNA-ploidy pattern, Ki-67 index or frequencies of mitosis and apoptosis between the two groups. In deeply invasive tumours (involving the muscularis propria or deeper), p53-positive ones were rare in the EBV-encoded small RNA (EBER)-positive group and very common in the EBER-negative group, while in superficial tumours, around one-half of them were p53-positive in both groups. In p53-positive superficial tumours, p21(WAF1)-positive ones accounted for 80% in the EBER-positive group and were scarce in the EBER-negative group. It is thus possible that p53 immunoreactivity reflects the overexpression of wild-type p53 and the stabilization of mutant p53 in the EBER-positive and -negative groups, respectively. The role of p53 in tumour development seems to be smaller in EBER-positive than in -negative gastric carcinomas.

The pain of not being recognised in the fight against Alzheimer's disease is a question of non-sharing rarely considered by medicine, but which is nevertheless a significant issue for the patient and carer. This anxiety relating to the fracture of the being emerges when being faced with the ordeal of Alzheimer's disease becomes a source of unfathomable helplessness. It is the question facing the caregiver with the arrival of the fateful stage of the non-recognition of the carer by the patient and the consequent lack of acknowledgement on the part of his or her family, and the non-sharing of his or her suffering which is not easily accepted by society.

Animals switch gaits according to locomotor speed. In terrestrial locomotion, gaits have been defined according to footfall patterns or differences in center of mass (COM) motion, which characterizes mechanisms that are more general and more predictive than footfall patterns. This has generated different variables designed primarily to evaluate steady-speed locomotion, which is easier to standardize in laboratory conditions. However, in the ecology of an animal, steady-state conditions are rare and the ability to accelerate, decelerate and turn is essential. Currently, there are no data available that have tested whether COM variables can be used in accelerative or decelerative conditions. This study used a data set of kinematics and kinetics of horses using three gaits (walk, trot, canter) to evaluate the effects of acceleration (both positive and negative) on commonly used gait descriptors. The goal was to identify variables that distinguish between gaits both at steady state and during acceleration/deceleration. These variables will either be unaffected by acceleration or affected by it in a predictable way. Congruity, phase shift and COM velocity angle did not distinguish between gaits when the dataset included trials in unsteady conditions. Work (positive and negative) and energy recovery distinguished between gaits and showed a clear relationship with acceleration. Hodographs are interesting graphical representations to study COM mechanics, but they are descriptive rather than quantitative. Force angle, collision angle and collision fraction showed a U-shaped relationship with acceleration and seem promising tools for future research in unsteady conditions.

Simple copper salts are known to denature poly d(GC). On the other hand, copper complexes of substituted 1,4,7,10,13-pentaazacyclohexadecane-14,16-dione are able to convert the right-handed B form of the same DNA sequence to the corresponding left-handed Z form. A research program was started in order to understand why Cu(II) as an aquated ion melts DNA and induces the conformational change to Z-DNA in the form of an azamacrocyclic complex. In this paper, we present a continuous wave and pulse electron paramagnetic resonance study of the mononucleotide model system Cu(II)-guanosine 5'-monophosphate . Pulse EPR methods like electron-nuclear double resonance and hyperfine sublevel correlation spectroscopy provide unique information about the electronic and geometric structure of this model system through an elaborate mapping of the hyperfine and nuclear quadrupole interactions between the unpaired electron of the Cu(II) ion and the magnetic nuclei of the nucleotide ligand. It was found that the Cu(II) ion is directly bound to N7 of guanosine 5'-monophosphate and indirectly bound via a water of hydration to a phosphate group. This set of experiments opens the way to more detailed structural characterization of specifically bound metal ions in a variety of nucleic acids of biological interest, in particular to understand the role of the metal-(poly)nucleotide interaction.

Lithium salts are the recommended first-line treatment (gold standard) in national and international treatment guidelines for acute and maintenance treatment of affective disorders, such as bipolar disorders. Lithium has also been shown to have a unique protective effect against suicide in patients suffering from affective disorders. Despite the well-known acute and long-term adverse effects lithium therapy can be safely administered if patients are properly educated and carefully monitored. A recent study from France now shows that patients with severely impaired renal function who had been treated with lithium salts for more than 10 years could have an increased risk for kidney tumors (benign and malignant). This resulted in an adjustment concerning information within the package leaflet by European authorities. The authors of this article reflect the currently available data in order to better understand and handle this new finding and to warn about uncritical reactions including withdrawal of lithium in successfully treated patients. This article provides clinical recommendations to provide further insight relating to the risk of kidney cancer in long-term lithium therapy.

Radiotherapy plays an important role in the treatment of early-stage Hodgkin lymphoma, but late toxicities such as cardiovascular disease and second malignancy are a major concern. Our aim was to evaluate the potential of deep inspiration breath-hold (DIBH) and intensity-modulated radiotherapy (IMRT) to reduce cardiac dose from mediastinal radiotherapy. A 24 year-old male with early-stage bulky mediastinal Hodgkin lymphoma received involved-site radiotherapy as part of a combined modality programme. Simulation was performed in free breathing (FB) and DIBH. The target and organs at risk were contoured on both datasets. Free breathing-3D conformal (FB-3DCRT), DIBH-3DCRT, FB-IMRT and DIBH-IMRT were compared with respect to target coverage and doses to organs at risk. A 'butterfly' IMRT technique was used to minimise the low-dose bath. In our patient, both DIBH (regardless of mode of delivery) and IMRT (in both FB and DIBH) achieved reductions in mean heart dose. DIBH improved all lung parameters. IMRT reduced high dose (V20), but increased low dose (V5) to lung. DIBH-IMRT was chosen for treatment delivery. Advanced radiotherapy techniques have the potential to further optimise the therapeutic ratio in patients with mediastinal lymphoma. Benefits should be assessed on an individualised basis.

Primary cilia are solitary, generally non-motile, hair-like protrusions that extend from the surface of cells between cell divisions. Their antenna-like structure leads naturally to the assumption that they sense the surrounding environment, the most common hypothesis being sensation of mechanical force through calcium-permeable ion channels within the cilium. This Ca(2+)-responsive mechanosensor hypothesis for primary cilia has been invoked to explain a large range of biological responses, from control of left-right axis determination in embryonic development to adult progression of polycystic kidney disease and some cancers. Here we report the complete lack of mechanically induced calcium increases in primary cilia, in tissues upon which this hypothesis has been based. We developed a transgenic mouse, Arl13b-mCherry-GECO1.2, expressing a ratiometric genetically encoded calcium indicator in all primary cilia. We then measured responses to flow in primary cilia of cultured kidney epithelial cells, kidney thick ascending tubules, crown cells of the embryonic node, kinocilia of inner ear hair cells, and several cell lines. Cilia-specific Ca(2+) influxes were not observed in physiological or even highly supraphysiological levels of fluid flow. We conclude that mechanosensation, if it originates in primary cilia, is not via calcium signalling.

To elucidate the role of neuropeptide Y (NPY)-Y1 receptor (Y1-R) in food intake, energy expenditure, and other possible functions, we have generated Y1-R-deficient mice (Y1-R-/-) by gene targeting. Contrary to our hypothesis that the lack of NPY signaling via Y1-R would result in impaired feeding and weight loss, Y1-R-/- mice showed a moderate obesity and mild hyperinsulinemia without hyperphagia. Although there was some variation between males and females, typical characteristics of Y1-R-/- mice include: greater body weight (females more than males), an increase in the weight of white adipose tissue (WAT) (approximately 4-fold in females), an elevated basal level of plasma insulin (approximately 2-fold), impaired insulin secretion in response to glucose administration, and a significant changes in mitochondrial uncoupling protein (UCP) gene expression (up-regulation of UCP1 in brown adipose tissue and down-regulation of UCP2 in WAT). These results suggest either that the Y1-R in the hypothalamus is not a key molecule in the leptin/NPY pathway, which controls feeding behavior, or that its deficiency is compensated by other receptors, such as NPY-Y5 receptor. We believe that the mild obesity found in Y1-R-/- mice (especially females) was caused by the impaired control of insulin secretion and/or low energy expenditure, including the lowered expression of UCP2 in WAT. This model will be useful for studying the mechanism of mild obesity and abnormal insulin metabolism in noninsulin-dependent diabetes mellitus.

The development of drugs for neglected infectious diseases often uses parasite-specific enzymes as targets. We here demonstrate that parasite enzymes with highly conserved human homologs may represent a promising reservoir of new potential drug targets. The cyclic nucleotide-specific phosphodiesterases (PDEs) of Trypanosoma brucei, causative agent of the fatal human sleeping sickness, are essential for the parasite. The highly conserved human homologs are well-established drug targets. We here describe what is to our knowledge the first pharmacological validation of trypanosomal PDEs as drug targets. High-throughput screening of a proprietary compound library identified a number of potent hits. One compound, the tetrahydrophthalazinone compound A (Cpd A), was further characterized. It causes a dramatic increase of intracellular cyclic adenosine monophosphate (cAMP). Short-term cell viability is not affected, but cell proliferation is inhibited immediately, and cell death occurs within 3 days. Cpd A prevents cytokinesis, resulting in multinucleated, multiflagellated cells that eventually lyse. These observations pharmacologically validate the highly conserved trypanosomal PDEs as potential drug targets.

To increase the nisin synthesizing activity of the natural strain Lactococcus lactis subsp. lactis 119 isolated from sour milk, UV irradiation in different doses was used followed by isolation of productive clones. The highest mutation effect was observed with the dose of 76,000 erg/mm2, when 11% of the cells increased their productivity by 12.8% at the minimum survival rate. Two-step UV irradiation and adaptive selection on the nisin-contaning medium provided isolation of a strain with the activity 42.6% higher than that of the initial strain (3850 IU/ml). Natural and UV-induced variability of the strain by the nisin synthesis, growth rate, carbohydrate consumption and sensitivity to antibiotics of various groups were studied.

BACKGROUND AND AIMS: Chronic ethanol consumption is associated with hypertension and atherosclerosis. Vascular oxidative stress is described as an important mechanism whereby ethanol predisposes to atherosclerosis. We hypothesized that nebivolol would prevent ethanol-induced hypertension and vascular oxidative stress.METHODS: Male Wistar rats were treated with ethanol 20% (vol./vol.) or nebivolol (10 mg/kg/day, p. o., gavage), a selective â1-adrenergic receptor antagonist.RESULTS: Ethanol-induced increase in blood pressure and in the circulating levels of adrenaline and noradrenaline was prevented by nebivolol. Similarly, nebivolol prevented ethanol-induced increase in plasma levels of renin, angiotensin I and II. Chronic ethanol consumption increased the aortic levels of superoxide anion (O2-), thiobarbituric acid reactive species (TBARS) as well as the expression of Nox1 and nitrotyrosine immunostaining in the rat aorta. Treatment with nebivolol prevented these responses. The decrease in aortic levels of nitrate/nitrite (NOx) induced by ethanol was prevented by the treatment with nebivolol. Finally, nebivolol attenuated ethanol-induced increase in phenylephrine- and noradrenaline-induced contraction of endothelium-intact and endothelium-denuded aortic rings.CONCLUSIONS: The novelty of our study is that nebivolol prevented ethanol-induced hypertension and vascular oxidative stress. Additionally, we showed that the sympathetic nervous system (SNS) and the renin-angiotensin system (RAS) are important endogenous mediators of the cardiovascular effects of ethanol.

Selected qualitative and biochemical parameters were determined in cryopreserved semen used for artificial insemination, sampled from 120 bulls reared at the Animal Breeding and Insemination Center in Bydgoszcz. The total average motility of the analyzed sperm samples was determined at 62.51%. The percentage of motile spermatozoa displaying progressive forward motility was 21.65%. Analyzed samples were characterized by a high percentage of sperm cells with a intact plasma membrane (71.21%) and active mitochondria (71.32%). High efficiency of the enzymatic antioxidant system of the evaluated sperm cells was demonstrated by high activity of CAT, GPx and SOD (494.37, 2847.83 and 5.31U/1x10(9) spermatozoa, respectively) values and low values of the DNA Fragmentation Index (9.32). The results of the study, obtained with the involvement of advanced analytical methods, indicate a high fertilizing capability of the analyzed sperm samples.

Self-inflicted injuries are probably more common than is generally appreciated. Three patients with self-inflicted hand injuries are described. A delay in making the correct diagnosis resulted in severe hand disability in 2 of the patients. Early diagnosis of this entity, combined with prompt psychotherapy, prevented disability in the third. The pertinent literature is reviewed and the importance of early diagnosis and psychotherapy is stressed.

OBJECTIVE: To rapidly deploy a digital patient-facing self-triage and self-scheduling tool in a large academic health system to address the COVID-19 pandemic.MATERIALS AND METHODS: We created a patient portal-based COVID-19 self-triage and self-scheduling tool and made it available to all primary care patients at the University of California, San Francisco Health, a large academic health system. Asymptomatic patients were asked about exposure history and were then provided relevant information. Symptomatic patients were triaged into 1 of 4 categories-emergent, urgent, nonurgent, or self-care-and then connected with the appropriate level of care via direct scheduling or telephone hotline.RESULTS: This self-triage and self-scheduling tool was designed and implemented in under 2 weeks. During the first 16 days of use, it was completed 1129 times by 950 unique patients. Of completed sessions, 315 (28%) were by asymptomatic patients, and 814 (72%) were by symptomatic patients. Symptomatic patient triage dispositions were as follows: 193 emergent (24%), 193 urgent (24%), 99 nonurgent (12%), 329 self-care (40%). Sensitivity for detecting emergency-level care was 87.5% (95% CI 61.7-98.5%).DISCUSSION: This self-triage and self-scheduling tool has been widely used by patients and is being rapidly expanded to other populations and health systems. The tool has recommended emergency-level care with high sensitivity, and decreased triage time for patients with less severe illness. The data suggests it also prevents unnecessary triage messages, phone calls, and in-person visits.CONCLUSION: Patient self-triage tools integrated into electronic health record systems have the potential to greatly improve triage efficiency and prevent unnecessary visits during the COVID-19 pandemic.

Margination refers to the migration of particles toward blood vessel walls during blood flow. Understanding the mechanisms that lead to margination will aid in tailoring the attributes of drug-carrying particles for effective drug delivery. Most previous studies evaluated the margination propensity of these particles via an adhesion mechanism, i.e., by measuring the number of particles that adhered to the channel wall. Although particle adhesion and margination are related, adhesion also depends on other factors. In this study, we quantified the margination propensity of particles of varying diameters (0.53, 0.84, and 2.11 ìm) and apparent wall shear rates (30, 61, and 121 s-1) by directly tracking fluorescent particles flowing through a microfluidic channel. The margination parameter, M, is defined as the total number of particles found within the cell-free layers normalized by the total number of particles that passed through the channel. In this study, an M-value of 0.2 indicated no margination, which was observed for all particle sizes in water. In the case of blood, larger particles were found to have higher M-values and thus marginated more effectively than smaller particles. The corresponding M-values at the device outlet were 0.203, 0.223, and 0.285 for 0.53-, 0.84-, and 2.11-ìm particles, respectively. At the inlet, the M-values for all particle sizes in blood were <0.2, suggesting that non-fully-developed flow and constriction may lead to demargination. For particle velocities transverse to the flow direction (vy), all particle sizes showed a larger standard deviation of vy as well as a higher effective diffusivity when the particles were suspended in blood relative to water. These higher values are attributed to collisions between the blood cells and particles, further supporting recent simulation results. In terms of flow rates, for a given particle size, the higher the flow rate, the higher the M-value.

We studied antipyrine clearance (APC1) in 19 healthy controls, 10 cirrhotic patients, 20 patients undergoing partial hepatectomy for liver tumors (14 with cirrhosis and 6 without cirrhosis). The aim of the study was to evaluate if the test represents a measure of the residual hepatic function and a useful index of surgical risk in cirrhotic patients following partial hepatectomy. The APC1 was significantly reduced in cirrhotic patients ws healthy controls. It is reduced following partial hepatectomy in cirrhotic patients (p less than 0.001) and in non cirrhotic patients (n.s.). The APC1 was found to be related with the plasma level of albumin and pseudocholinesterase; it also was related with the Pugh's score for hepatic function. The APC1 is a satisfactory index of residual hepatic function. On the other hand it does not provide more useful information than the Pugh's score for surgical risk in liver resection.

Live probiotic bacteria are effective in reducing gut permeability and inflammation. We have previously shown that probiotics release peptide bioactive factors that modulate epithelial resistance in vitro. The objectives of this study were to determine the impact of factors released from Bifidobacteria infantis on intestinal epithelial cell permeability and tight junction proteins and to assess whether these factors retain their bioactivity when administered to IL-10-deficient mice. B. infantis conditioned medium (BiCM) was applied to T84 human epithelial cells in the presence and absence of TNF-alpha and IFN-gamma. Transepithelial resistance (TER), tight junction proteins [claudins 1, 2, 3, and 4, zonula occludens (ZO)-1, and occludin] and MAP kinase activity (p38 and ERK) were examined. Acute effects of BiCM on intestinal permeability were assessed in colons from IL-10-deficient mice in Ussing chambers. A separate group of IL-1-deficient mice was treated with BiCM for 4 wk and then assessed for intestinal histological injury, cytokine levels, epithelial permeability, and immune response to bacterial antigens. In T84 cells, BiCM increased TER, decreased claudin-2, and increased ZO-1 and occludin expression. This was associated with enhanced levels of phospho-ERK and decreased levels of phospho-p38. BiCM prevented TNF-alpha- and IFN-gamma-induced drops in TER and rearrangement of tight junction proteins. Inhibition of ERK prevented the BiCM-induced increase in TER and attenuated the protection from TNF-alpha and IFN-gamma. Oral BiCM administration acutely reduced colonic permeability in mice whereas long-term BiCM treatment in IL-10-deficient mice attenuated inflammation, normalized colonic permeability, and decreased colonic and splenic IFN-gamma secretion. In conclusion, peptide bioactive factors from B. infantis retain their biological activity in vivo and are effective in normalizing gut permeability and improving disease in an animal model of colitis. The effects of BiCM are mediated in part by changes in MAP kinases and tight junction proteins.

The prevalence of serum thyroid-stimulating immunoglobulins (T.S.I.) in a variety of thyroid diseases was determined in 96 patients and 35 normal controls. Significantly elevated levels of T.S.I. were found not only in patients with Graves' disease and Hashimoto's thyroiditis but also in those with non-toxic and toxic multinodular goitre, whereas patients with a single autonomously functioning thyroid nodule, with subacute thyroiditis, and with "hyperthyroiditis" had levels which did not differ from those in the controls. We postulate that non-toxic multinodular goitre, like Graves' disease, may result from increased circulating T.S.I., which in some cases may be present in sufficient concentration to cause thyrotoxicosis.

Cytokine responses in the Japanese pufferfish (Takifugu rubripes) head kidney (HK) cells to heat-killed lactic acid bacteria probiotics isolated from the Mongolian dairy products were investigated by transcriptomic examination. The HK cells were incubated with two heat-killed bacteria, namely Lactobacillus paracasei spp. paracasei (strain 06TCa22) and L. plantarum (strain 06CC2) and the responses of 16 cytokine genes at 0 (control), 1, 4, 8, 12, 24 and 48 h post-stimulation were assayed by multiplex RT-PCR analysis (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter, Inc.). The 16 genes included in the assay were pro-inflammatory cytokines (IL-1â, IL-6, IL-17A/F-3, TNF-á and TNF-N), cell-mediated immune regulators (IL-12p35, IL-12p40 and IL-18), antiviral (I-IFN-1 and IFN-ã) and other regulatory (IL-2, IL-7, IL-15, IL-21, IL-10 and TGF-â1) cytokines. Despite the differences in the transcriptional profiles, expression of all the cytokines tested here was significantly elevated by both the probiotic bacterial stimulants compared with the unstimulated control. Therefore, this in vitro study has demonstrated the modulation of cytokine defense mechanisms in the HK cells by the two heat-killed probiotics indicating their potentiality as novel immunostimulants to fish. However, strain-dependent varied expression of important cytokines (cell-mediated immune regulators, antiviral and anti-inflammatory cytokines) suggests better efficacy of L. paracasei spp. paracasei strain as fish immunostimulant. Further in vivo studies to elucidate the cytokine regulation networks will validate our present observations. A careful evaluation of ant-inflammatory properties may be undertaken using single strain to affirm the immunostimulatory capability. Moreover, application timings and frequency to assess the longevity of immunostimulant effects and to make the application cost-effective need to be evaluated before any practical use in aquaculture.

A major problem in camel productivity is the high mortality rate of camel calves in the first 3 months. The causes for mortality are mainly poor management practice and infectious diseases. The purpose of this research, carried out on a ranch in Kenya, was to determine the immunoglobulin G (IgG) concentration in camel colostrum as well as the extent of the calves' passive immunization by maternal antibodies. IgG concentration in colostrum and in the serum of the calf were measured during the first 3 d of life. Evaluation was carried out by comparing the respective values with those for horses and cattle. The average IgG concentration in the camel colostrum was higher than that found in literature for horses and cattle. IgG concentration in the serum of the camel calves reached its maximum 24 h after birth. In 39% of the examined calves, this maximum concentration was below 4 g/l, which is considered to be the critical value in horses and cattle. 61% of the calves achieved an IgG concentration of over 4 g/l. Since there is no correlation between IgG level in colostrum and early mortality, the results indicate that low colostrum intake during the first 24 h of life and not low IgG concentration in colostrum is presumably one of the main causes of early calf mortality. Therefore, it was recommended that the care of the newborn calves by herdsmen should be improved.

Epistatic interactions between mutant sites in the same protein can exert a strong influence on pathways of molecular evolution. We performed protein engineering experiments that revealed pervasive epistasis among segregating amino acid variants that contribute to adaptive functional variation in deer mouse hemoglobin (Hb). Amino acid mutations increased or decreased Hb-O2 affinity depending on the allelic state of other sites. Structural analysis revealed that epistasis for Hb-O2 affinity and allosteric regulatory control is attributable to indirect interactions between structurally remote sites. The prevalence of sign epistasis for fitness-related biochemical phenotypes has important implications for the evolutionary dynamics of protein polymorphism in natural populations.

It is a challenge for the human body to maintain stable blood pressure while standing. The body's failure to do so can lead to dizziness or even fainting. For decades it has been postulated that the vestibular organ can prevent a drop in pressure during a position change--supposedly mediated by reflexes to the cardiovascular system. We show--for the first time--a significant correlation between decreased functionality of the vestibular otolith system and a decrease in the mean arterial pressure when a person stands up. Until now, no experiments on Earth could selectively suppress both otolith systems; astronauts returning from space are a unique group of subjects in this regard. Their otolith systems are being temporarily disturbed and at the same time they often suffer from blood pressure instability. In our study, we observed the functioning of both the otolith and the cardiovascular system of the astronauts before and after spaceflight. Our finding indicates that an intact otolith system plays an important role in preventing blood pressure instability during orthostatic challenges. Our finding not only has important implications for human space exploration; they may also improve the treatment of unstable blood pressure here on Earth.

Therapeutic methods for treating poisoned men are based on the principles of intensive care therapy. The elimination of toxins depends on the circulatory circumstances; an enhancement of poison elimination can be achieved in providing a normal circulation. In the treatment of most cases of poisoning gastric and gut lavage will be sufficient. An extracorporeal toxin elimination with hemodialysis or hemoperfusion will be reserved for special cases, because of increased technical and personal expense and a higher complication rate, compared to other methods of treatment in poisoning. Forced diuresis is a simple method, but the efficacy is reduced to a small number of substances. Also reduced to certain toxins is the method of elimination with hyperventilation.

Traditional approaches to human causal reasoning assume that the perception of temporal order informs judgments of causal structure. In this article, we present two experiments in which people followed the opposite inferential route: Perceptual judgments of temporal order were instead influenced by causal beliefs. By letting participants freely interact with a software-based "physics world," we induced stable causal beliefs that subsequently determined participants' reported temporal order of events, even when this led to a reversal of the objective temporal order. We argue that for short timescales, even when temporal-resolution capabilities suffice, the perception of temporal order is distorted to fit existing causal beliefs.

BACKGROUND: In all IVF programmes, some patients fail to achieve an ongoing pregnancy, even after numerous embryo transfer procedures. An unfavourable environment within the uterus might be a contributory factor to such recurrent implantation failure. This question was addressed by measuring cytokine concentrations and matrix metalloproteinase activities in fluid derived from uterine irrigation of such patients.METHODS AND RESULTS: The uterine cavities of 22 patients who had previously undergone embryo transfer of at least 10 embryos without ongoing pregnancy were irrigated during the luteal phase. The resultant fluid was assayed for the concentration of interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, leukaemia inhibitory factor (LIF), IL-10 and matrix metalloproteinase (MMP) -2 and -9 activity. The results were compared with those of a control population of women known to be previously fertile (n = 16) and also with women with recurrent spontaneous abortion (n = 13). In the recurrent implantation failure group, the MMP score and IL-1beta concentration were significantly higher than those in the control group, whereas concentrations of IFN-gamma and IL-10 were significantly lower.CONCLUSIONS: In IVF patients with recurrent implantation failure, an altered pattern of intra-uterine cytokine concentration and MMP activity was observed.

Motile cilia and flagella exhibit many waveforms as outputs of dynein activation sequences on the highly conserved axoneme. Motility change of sperm in the reproductive tract is difficult to study and remains an important area of investigation. Sperm typically execute a sinusoidal waveform. Increased viscosity in the medium induces somewhat unusual arc-line and helical waveforms in some sperm. However, whether the latter two waveforms occur in vivo is not known. Using green fluorescence protein imaging, we show that Drosophila sperm in the uterus move in circular foci via arc-line waves, predominantly in a tail-leading orientation. From the uterus, a small fraction of the sperm enters the seminal receptacle (SR) in parallel formations. After sperm storage and coincident with fertilization of the egg, the sperm exit the SR via head-leading helical waves. Consistent with the observed bidirectional movements, the sperm show the ability to propagate both base-to-tip and tip-to-base flagellar waves. Numerous studies have shown that sperm motility is regulated by intraflagellar calcium concentrations; in particular, the Pkd2 calcium channel has been shown to affect sperm storage. Our analyses here suggest that Pkd2 is required for the sperm to adopt the correct waveform and movement orientation during SR entry. A working model for the sperm's SR entry movement is proposed.

European eels are dangerously threatened with extinction. Recent advances tend to show that pollution could, in addition to other already identified factors, contribute to this drama. In a previous report, cadmium (Cd) pre-exposure was found to strongly stimulate the pituitary-liver-gonad axis of maturing female silver eels, leading, lastly, to oocytes atresia and eels mortality. The present work was performed to get more insights into the effects of Cd preexposure on eels' ovaries. The transcription levels of various genes involved in mitochondrial metabolism, in the cellular response to metal (metallothioneins, MTs) and oxidative stress (catalase, CAT) were investigated. Our results show that ovarian growth is associated with an up-regulation of mitochondrial genes. However, Cd pre-exposure was found to significantly impair this up-regulation. Such findings could explain, at least in part, why oocytes of Cd pre-contaminated eels could not reach final maturation. Concerning MTs, despite the end of the experiment was marked by a strong increase in their gene transcription levels in both eel groups, MTs protein content was found to increase only in the case of Cd pre-contaminated eels. Since this increase in MTs protein content was associated with a massive entry of Cd in gonads, our findings suggest that MTs mRNA, that are normally accumulated in oocytes to cope with the future needs, can be activated and translated in response to Cd exposure.

Mutations that uncouple glucose transport from phosphorylation were isolated in plasmid-encoded Escherichia coli enzyme IIGlc of the phosphoenolpyruvate-dependent phosphotransferase system (PTS). The uncoupled enzymes IIGlc were able to transport glucose in the absence of the general phosphoryl-carrying proteins of the PTS, enzyme I and HPr, although with relatively low affinity. Km values of the uncoupled enzymes IIGlc for glucose ranged from 0.5 to 2.5 mM, 2 orders of magnitude higher than the value of normal IIGlc. Most of the mutant proteins were still able to phosphorylate glucose and methyl alpha-glucoside (a non-metabolizable glucose analog specific for IIGlc), indicating that transport and phosphorylation are separable functions of the enzyme. Some of the uncoupled enzymes IIGlc transported glucose with a higher rate and lower apparent Km in a pts+ strain than in a delta ptsHI strain lacking the general proteins enzyme I and HPr. Since the properties of these uncoupled enzymes IIGlc in the presence of PTS-mediated phosphoryl transfer resembled those of wild-type IIGlc, these mutants appeared to be conditionally uncoupled. Sequencing of the mutated ptsG genes revealed that all amino acid substitutions occurred in a hydrophilic segment within the hydrophobic N-terminal part of IIGlc. These results suggest that this hydrophilic loop is involved in binding and translocation of the sugar substrate.

The role of the autonomic nervous system on the pulmonary vascular response to hypoxia was studied in anesthetized open-chest dogs. Acute hypoxia elicited reversible decreases in the slope of the instantaneous diameter-pressure relationship of the main pulmonary artery. These decrements in vascular compliance during hypoxia were prevented by either bilateral thoracic sympathectomy or by section of the cervical vagus and glossopharyngeal nerves and were reproduced by bolus injections of lobeline (0.1 mg) at the root of the aorta. Using separation of the airways with a tracheal divider, the effects of systemic and of alveolar hypoxia on vascular resistance of the left caudal lobe were compared. Systemic hypoxia in the absence of alveolar hypoxia had no effect on pulmonary vascular resistance. Alveolar hypoxia elicited reversible increases in vascular resistance that were not prevented by sympathectomy. It is concluded that at least two vascular sites of action and two mechanisms are involved in the pulmonary vascular response to hypoxia.

A site-directed mutagenesis study on the conserved subunit I histidines of the cytochrome bo complex in Escherichia coli identified ligands of the low spin heme B and CuB centers; however, the assignment of the proximal ligand of the high spin heme O was ambiguous (Minagawa, J., Mogi, T., Gennis, R. B., and Anraku, Y. (1992) J. Biol. Chem. 267, 2096-2104). We have extended this work and characterized the metal centers in the purified histidine mutant oxidases by optical, EPR, and resonance Raman spectroscopies and by biochemical analysis. We found that the H284A and H333A oxidases contain two heme B molecules, which exhibit the gz = 2.99 low spin and cyanide-sensitive g perpendicular = 6 high-spin EPR signals, whereas the H419A oxidase contains only low spin heme B, which shows the gz component in a considerably higher magnetic field at g = 2.92. The CuB center was partially retained in the H284A oxidase but was almost completely lost in the H333A and H419A oxidases. Thus, we concluded that His419 is the proximal ligand of the high spin heme O and that His284 is located at the distal side of the high spin heme O. His284 plays an indispensable role in maintaining the structure of the Feo-CuB binuclear site suitable for the exogenous ligand bindings in the reduced state, since its substitution eliminated the CO binding activity. In addition, we found that His106 and His421 are in fact the axial ligands of the low spin heme B, although the H421A mutation perturbed the binuclear metal center seriously. Based upon experimental results with isotopic substitutions on iron in the oxidases, we assigned the Raman band at 208 cm-1 to the iron-histidine stretching mode (vFc(2+)-N(His)) of the wild-type ferrous cytochrome o, which was upshifted slightly by a loss of the CuB center in the H333A oxidase. A molecular structure of the metal centers and a possible mechanism of the electron transfer-coupled proton pumping in the cytochrome bo complex are proposed on the basis of our present findings.

The chromatin structure specific to the SV40 late transcription elongation complex as well as the occupancy of several sites that bind transcription factors have been examined. These features have been determined by assessing blockage to restriction enzyme digestion. Cleavage specific to the elongation complex has been quantified using ternary complex analysis. This method involves radioactively labeling the complex by in vitro transcription followed by determining the extent of linearization by electrophoresis in an agarose gel. It was found that not only is the origin region devoid of nucleosomes, but there is also no stable factor occupancy at the BglI, SphI, KpnI and MspI restriction enzyme sites within this region. Thus these sites were cleaved to a high degree, meaning that the binding sites for a number of transcription factors, including OBP/TEF-1, TBP, DAP, as well as a proposed positioned nucleosome, are unoccupied in the native viral transcription elongation complex. The absence of these trans-acting factors from their respective binding sites in the elongation complex indicates that they bind only transiently, possibly cycling on and off during the transcription cycle. This finding implies that various forms of transcription complex are assembled and disassembled during transcription and thus supports a 'hit-and-run' model of factor function.

This paper presents IEC/SC 45B 'Radiation protection instrumentation' and its standards for individual monitoring of ionising radiation: IEC 61526 Ed. 3 for active personal dosemeters and IEC 62387-1 for passive integrating dosimetry systems. The transposition of these standards as CENELEC (European) standards is also discussed together with the collaboration between IEC/SC 45B and ISO/TC 85/SC 2.

PURPOSE: To evaluate if cardiac magnetic resonance elastography (MRE) can measure increased stiffness in patients with cardiac amyloidosis. Myocardial tissue stiffness plays an important role in cardiac function. A noninvasive quantitative imaging technique capable of measuring myocardial stiffness could aid in disease diagnosis, therapy monitoring, and disease prognostic strategies. We recently developed a high-frequency cardiac MRE technique capable of making noninvasive stiffness measurements.MATERIALS AND METHODS: In all, 16 volunteers and 22 patients with cardiac amyloidosis were enrolled in this study after Institutional Review Board approval and obtaining formal written consent. All subjects were imaged head-first in the supine position in a 1.5T closed-bore MR imager. 3D MRE was performed using 5 mm isotropic resolution oblique short-axis slices and a vibration frequency of 140 Hz to obtain global quantitative in vivo left ventricular stiffness measurements. The median stiffness was compared between the two cohorts. An octahedral shear strain signal-to-noise ratio (OSS-SNR) threshold of 1.17 was used to exclude exams with insufficient motion amplitude.RESULTS: Five volunteers and six patients had to be excluded from the study because they fell below the 1.17 OSS-SNR threshold. The myocardial stiffness of cardiac amyloid patients (median: 11.4 kPa, min: 9.2, max: 15.7) was significantly higher (P = 0.0008) than normal controls (median: 8.2 kPa, min: 7.2, max: 11.8).CONCLUSION: This study demonstrates the feasibility of 3D high-frequency cardiac MRE as a contrast-agent-free diagnostic imaging technique for cardiac amyloidosis.LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2017;46:1361-1367.

BACKGROUND: Measurement of volumetric changes with MR might be a useful surrogate endpoint for clinical trials in frontotemporal lobar degeneration (FTLD). Because there is only limited longitudinal imaging data currently available, we measured the rate of change over 1 year of whole brain volume (WBV) and ventricular volume (VV) in patients with FTLD.METHODS: Subjects with an FTLD cognitive syndrome were recruited from five centers using standard clinical diagnostic criteria for behavioral variant frontotemporal dementia (bvFTD), progressive nonfluent aphasia (PNFA), semantic dementia (SMD), and progressive logopenic aphasia. Structural brain imaging, using three-dimensional T1-weighted sequences at 1.5 teslas, and cognitive, behavioral, and functional assessments were performed at baseline and approximately 1 year later. The boundary shift integral algorithm was used to determine change in WBV and VV.RESULTS: There were 76 patients (mean age 64 years; 41 men and 35 women) who had usable baseline and annual scans. The group-wise annualized change was -1.62% (SD 1.03, range +0.69 to -3.6) for WBV and 11.6% (SD 5.9, range -1.3 to 23.9) for VV. Rates of change were similar among bvFTD, PNFA, and SMD groups. Longitudinal changes in WBV and VV were correlated with decline on clinical global and cognitive measures.CONCLUSIONS: Multicenter, serial measurements of whole brain volume (WBV) and ventricular volume (VV) from magnetic resonance scans were feasible in patients with frontotemporal lobar degeneration (FTLD). Using WBV or VV as outcome measures would require recruiting (at 80% power) 139 or 55 subjects per group to detect a small (25%) or medium-sized (40%) effect in a randomized, placebo-controlled trial of a putative agent for FTLD.

Erv41p is a conserved integral membrane protein that is known to play a role in transport between the endoplasmic reticulum and Golgi apparatus, part of the early secretory pathway of eukaryotes. However, the exact function of the protein is not known, and it shares very low sequence identity with proteins of known structure or function. Here we present the structure of the full lumenal domain of Erv41p from Saccharomyces cerevisiae, determined by X-ray crystallography to a resolution of 2.0?. The structure reveals the protein to be composed predominantly of two large â-sheets that form a twisted â-sandwich. Comparison to structures in the Protein Data Bank shows that the Erv41p lumenal domain displays only limited similarity to â-sandwich domains of other proteins. Analysis of the surface properties of the protein identifies an extensive patch of negative electrostatic potential on the exposed surface of one of the â-sheets, which likely forms a binding site for a ligand or interaction partner. A predominantly hydrophobic region close to the membrane interface is identified as a likely site for protein-protein interaction. This structure, the first of Erv41p or any of its homologues, provides a new starting point for studies of the roles of Erv41p and its interaction partners in the eukaryotic secretory pathway.

This report further characterizes the intermediate metabolic effects of the psychotropic amphetamine derivative, 3,4-methylenedioxymethamphetamine (MDMA or "ecstasy"), on the activity of second messenger-dependent kinases. Previous work has demonstrated that two injections of MDMA (20 mg/kg) elicits a prolonged translocation of the calcium and phospholipid-dependent enzyme, protein kinase C (PKC) in rats. However, because MDMA has actions at the 5-HT transporter and 5-HT2A/2C receptors, our experiments were directed at uncovering which of these many sites may be involved in this second messenger dependent response. A single injection of MDMA produced a time- and dose-dependent increase in the density of cortical and hippocampal PKC (as measured by 3H-phorbol 12,13-dibutyrate (PDBu) binding sites. MDMA-mediated PKC translocation was long-lasting and remained above control (saline-treated rats) for up to 24 h after injection. This effect was mimicked by another substituted amphetamine, p-chloroamphetamine (pCA), but with a temporal-response curve that was to the left of MDMA's. However, pure uptake inhibitors like fluoxetine, cocaine, and the selective 5-HT2A/2C agonist, DOB, were unable to produce a long-lasting translocation of PKC binding sites in rat cortex. Fluoxetine, a selective serotonin uptake inhibitor (SSRI) and ketanserin a 5-HT2A antagonist, attenuated PKC translocation by MDMA with differing efficacies; however, both compounds completely prevented the loss of 5-HT uptake sties after multiple doses of MDMA. These results suggest that MDMA increases PKC translocation by two interrelated mechanisms that involve 5-HT2A/2C receptors and the 5-HT transporter. This pathway appears to include: (1) the drug binding to the 5-HT transporter, (2) the release of cytosolic 5-HT stores into the extracellular space, and (3) the activation of post-synaptic 5-HT2A/2C receptors linked to G-protein-mediated phospholipid hydrolysis.

OBJECTIVE: We reported that experimental autoimmune myocarditis (EAM) rats showed dramatic changes in ventricular action potential and enhanced arrhythmogenicity in the acute phase, but mechanisms for this are still unclear. To investigate the mechanisms of cardiac remodeling in acute myocarditis and subsequent heart failure, physiological and molecular changes were evaluated along the time course of EAM.METHODS: Six-week-old Lewis rats were immunized with porcine cardiac myosin. On days 14, 21, 35 and 60 after immunization, histology, hemodynamics and electrophysiological parameters (i.e., effective refractory period (ERP), monophasic action potential duration (MAPD) and PVC inducibility) were evaluated and compared with control rats. After these studies, the expression levels of Kv(+) and L-Ca(2+) channels, ion transporters and BNP expressions in the left ventricle were examined by quantitative real time RT-PCR and Western blot analysis.RESULTS: EAM rats showed acute myocarditis with massive infiltration of the mononuclear cells on days 14 and 21. Subsequently, a chronic dilated cardiomyopathy (DCM)-like structural change was observed on day 60. Hemodynamic parameters were worse in EAM than controls. ERP and MAPD were longer in EAM than controls, with a peak on day 21, which was parallel to PVC inducibility. mRNA levels of Kv4.2, Kv1.5, KChIP2, frequenin and SERCA2a, and the protein levels of Kv4.2 and Kv1.5, were reduced, especially in the acute phase.CONCLUSIONS: The initial reduction of Ito-related molecules, such as the expression levels of Kv4.2, 1.5, frequenin and KChIP2, and the prolongation of MAPD are considered to be a key mechanism of ventricular remodeling and cause the characteristic clinical findings in EAM in the acute inflammatory phase and chronic DCM phase.

Cytologic and morphometric investigations of the hypothesis, thyroid, thyroid and pancreas of rats after 19-d centrifugation at 2G revealed suppression of somatotropic hormone production by hypophyseal somatotrophs and insulin by pancreatic beta-cells, and intensification of thyrotrophic hormone production by hypophyseal thyreotrophs, thyroxin by thyreoid parenchyma and glucagon by pancreatic alpha-cells. These change in hormone production are viewed as an endocrine mechanism of the divergence of plastic processes and energy metabolism in animals subjected to chronic exposure to hypergravity.

A drop brought into contact with a nearby substrate can wet and spread against the substrate, forming a liquid bridge that exerts a capillary force. This force due to surface tension can be used to "grab" the substrate, pulling it toward the drop. "Wet" adhesion results from the parallel action of an array of small liquid bridges. The Florida palm beetle, Hemisphaerota cyanea, uses wet adhesion to defend itself against attacking predators by adhering to the palm leaf using an array of about 120,000 ìm-sized liquid bridges. The beetle's survival depends on the strength of adhesion which, in turn, depends on how liquid bridges break. Individual bridges break when they go unstable, according to their response curves. However, the ultimate strength of an individual bridge depends on the class of disturbances to which it is subjected, and it has been speculated that the beetle may have some control over this class. The authors experimentally study families of liquid bridge equilibria for their breaking limits when subjected to constant-length (L) and constant-force (F) disturbances. While to control constant-L disturbances is straightforward, to apply and control constant-F disturbances on a liquid bridge requires more ingenuity. The authors introduce an apparatus with a lever-arm and a ball-bearing slide. The authors then compare our experimentally measured bridge response curves to the force trace from experiments on the beetle (prior literature) to infer the mode of beetle detachment. Under normal loads, the beetle detaches as a constant-L instability for smaller loads and as a constant-F instability for larger loads. The beetle's ability to adjust the type and magnitude of loading in real time is not only crucial to its survival but has implications for the design of various engineering devices.

Many women whose premature infants are hospitalized in a newborn intensive care unit choose to express breast milk for their babies. Yet anxiety, fatigue, and emotional stress are powerful inhibitors of lactation. To facilitate the breast-feeding experience, intervention mothers were given a 20-minute audio cassette tape based on relaxation and visual imagery techniques. At a single follow-up expression of milk at the hospital approximately 1 week after enrollment, they expressed 63% more breast milk than a randomized group of control mothers. The fat content of the breast milk in the two groups was not significantly different. Among a small group of mothers whose infants were receiving mechanical ventilation, the increase in milk volume compared with that of control mothers was 121%. Longer-term effects of the relaxation/imagery approach (such as extending the duration of breast-feeding or reducing parental stress after hospital discharge) and the physiologic basis for the increased volume of expressed milk (improved milk production v more efficient milk ejection) are appropriate topics for future research.

Existing approaches to nonrigid structure from motion assume that the instantaneous 3D shape of a deforming object is a linear combination of basis shapes. These bases are object dependent and therefore have to be estimated anew for each video sequence. In contrast, we propose a dual approach to describe the evolving 3D structure in trajectory space by a linear combination of basis trajectories. We describe the dual relationship between the two approaches, showing that they both have equal power for representing 3D structure. We further show that the temporal smoothness in 3D trajectories alone can be used for recovering nonrigid structure from a moving camera. The principal advantage of expressing deforming 3D structure in trajectory space is that we can define an object independent basis. This results in a significant reduction in unknowns and corresponding stability in estimation. We propose the use of the Discrete Cosine Transform (DCT) as the object independent basis and empirically demonstrate that it approaches Principal Component Analysis (PCA) for natural motions. We report the performance of the proposed method, quantitatively using motion capture data, and qualitatively on several video sequences exhibiting nonrigid motions, including piecewise rigid motion, partially nonrigid motion (such as a facial expressions), and highly nonrigid motion (such as a person walking or dancing).

Exposure to environmental tobacco smoke is associated with increased respiratory morbidity in young children, but few studies have assessed such exposure objectively by urinary cotinine measurements. 501 children aged 1-5 years, a random 5% sample of children attending an outpatient clinic, were classified as exposed or non-exposed to environmental tobacco smoke with a cut-off of 10 ng cotinine per mg creatinine in urine. Exposed children were 3.5 times (95% CI 1.56-7.90, p < 0.0024) more likely to have increased respiratory morbidity (three or more episodes during the previous 12 months) than non-exposed children after adjustment for potential confounding factors.

A comparative ultrasonic, oculometric study was performed in negroes, West Nigeria (151 persons) and in caucasians, Denmark (88 persons). The negros have thinner lenses than caucasians, and overall ethnic difference of 0.12 mm was found. Male negroes have longer vitreous and axial lengths than male caucasians (VB 0.56 mm, AxL 0.30 mm).

Several arylhydrazonopropanedinitriles and an arylhydrazonopropane-diethyl ester (derivatives of well-known uncouplers of oxidative phosphorylation) have been studied with respect to their ability to radiosensitize E. coli B/r under oxic and hypoxic conditions. Of the compounds studied, 2-carboxyphenylhydrazono-propanedinitrile and 2-carboxyphenylhydrazonopropanediethylester were found to be the most efficient radiosensitizers under hypoxia, whilst the former compound was also found to provide radiosensitization under oxic conditions. Increased radiosensitization by 4-carboxyphenylhydrazonopropanedinitrile was observed on decreasing the pH of the irradiation incubation medium. The results are discussed with respect to the physicochemical properties of these compounds and their reactivity with thiols, for which data are presented.

Neurocognitive (NC) complications continue to afflict a substantial proportion of HIV-infected people taking effective antiretroviral therapy (ART). One contributing mechanism for this is antiretroviral neurotoxicity. Efavirenz (EFV) is associated with short-term central nervous system (CNS) toxicity, but less is known about its long-term effects. Our objective was to compare NC functioning with long-term use of EFV to that of a comparator, lopinavir-ritonavir (LPV/r), in a cohort of well-characterized adults. Four hundred forty-five patients were selected from the CNS HIV Antiretroviral Therapy Effects Research (CHARTER) cohort based on their use of either EFV (n = 272, mean duration 17.9 months) or LPV/r (n = 173, mean duration 16.4 months) and the lack of severe NC comorbidities. All patients had undergone standardized comprehensive NC testing. Univariable and multivariable analyses to predict NC outcomes were performed. Compared with LPV/r users, EFV users were more likely to be taking their first ART regimen (p < 0.001), were less likely to have AIDS (p < 0.001) or hepatitis C virus (HCV) coinfection (p < 0.05), had higher CD4+ T cell nadirs (p < 0.001), had lower peak (p < 0.001) and current (p < 0.001) plasma HIV RNA levels, and were less likely to have detectable HIV RNA in cerebrospinal fluid (CSF) (p < 0.001). Overall, EFV users had worse speed of information processing (p = 0.04), verbal fluency (p = 0.03), and working memory (p = 0.03). An interaction with HCV serostatus was present: Overall among HCV seronegatives (n = 329), EFV users performed poorly, whereas among HCV seropositives (n = 116), LPV/r users had overall worse performance. In the subgroup with undetectable plasma HIV RNA (n = 269), EFV users had worse speed of information processing (p = 0.02) and executive functioning (p = 0.03). Substantial differences exist between EFV and LPV/r users in this observational cohort, possibly because of channeling by clinicians who may have prescribed LPV/r to more severely ill patients or as second-line therapy. Despite these differences, EFV users had worse functioning in several cognitive abilities. A potentially important interaction was identified that could indicate that the NC consequences of specific antiretroviral drugs may differ based on HCV coinfection. The complexity of these data is substantial, and findings would best be confirmed in a randomized clinical trial.

In order to gain further insights of the regulatory mechanisms of human beta-like globin gene switch during erythroid development, we have studied protein-DNA interaction in vivo at the human adult beta and fetal gamma globin promoters and their upstream enhancer, 5'HS-2, in purified human adult erythroblasts, in which the beta, but not gamma or epsilon, globin gene is actively transcribing. This genomic footprinting analysis of adult erythroblasts was carried out in conjunction with those of different non-erythroid human tissues, an embryonic/fetal erythroid cell line K562, and several non-erythroid human cell lines. Protein-DNA binding in the beta globin promoter, in particular at the two CACC promoter boxes and the CCAAT box, is detectable only in the adult erythroblasts. As expected, the gamma globin promoters were bound with specific nuclear factors in the expressing K562 cells, but not in non-erythroid tissues or cell lines. Relatively weak protein binding could also be detected in the vicinities of the two CCAAT boxes of the inactive gamma globin promoters in the adult erythroblasts. Although the patterns of nuclear factor-DNA interaction in vivo at the NF-E2/AP1, GATA-1, and GT-I motifs of 5'HS-2 enhancer in adult erythroblasts are similar to those in K562 cells, we have identified a previously undetected factor-binding motif of 5'HS-2 that is protected only in the adult erythroblasts. This motif is identical in sequence to the 3'-CACC box of the human beta globin promoter, and it is well conserved at the same location among all mammalian 5'HS-2 enhancers, suggesting an important regulatory role of this element in human beta globin gene transcription in adult erythroblasts. All of the above four motifs of 5'HS-2 are free of nuclear factor binding in non-erythroid tissues, but two of them, NF-E2/AP1 and GT-I, are bound with factors in some non-erythroid cell lines but not in others. The functional implications of these genomic footprinting data and the tissue-specific CpG methylation patterns of the beta-like globin promoters we obtained by genomic sequencing are discussed in terms of positive and negative regulation of the human beta-like globin switch during erythroid development.

Patients with eating disorders (ED) show alterations in both their behaviour and their intake of food, frequently presenting nutritional and somatic affectations. Besides the classical forms such as anorexia or bulimia nervosa, there has recently been an increase in atypical or incomplete forms of ED, such as binge eating. Primary care (PC) services form the central and closest nucleus of health care for the individual and the family, where ED occur and leave their mark. This allows PC to provide an integral response at all levels of care for ED. Primary prevention at school, in the family and community is fundamental to avoiding its inception. Secondary prevention is based on early diagnosis and treatment of ED and favours a better prognosis of the illness. Tertiary prevention tries to reduce the serious consequences with rehabilitation measures to alleviate complications and avoid risk to life. Due to its complexity, these patients are afforded the attention of multidisciplinary teams of specialists with experience in treating this condition. In consultation with the team, the general practitioner should adopt a leading role at all levels of attention, as he/she is the link between the team, the family and the patient. This requires both regulated, specific training in the disease and the allocation of resources to carry it through. Putting into practice all these plans would allow us to give a positive answer to the question posed in the title of the present article.

The study reported here demonstrates the feasibility of presenting cognitive tests from the Cambridge Neuropsychological Test Automated Battery (CANTAB) to either marmosets or rhesus monkeys in their home cages. This location of testing offers opportunities for the measurement of additional indices, for example spontaneous behaviour (Prowse et al. 1995) and electrophysiology (Pearce et al. 1998) as well as facilitating repeated test presentation. Results from 12 marmosets and 4 rhesus monkeys which have completed several sequences of an eight-stage discrimination task involving simple discriminations, compound discriminations and reversals are reported. The paradigm developed has application in long-term studies. Tests from CANTAB have been used extensively in normal humans (Robbins et al. 1994) as well as a range of patient groups (Owen et al. 1992, Elliott et al. 1995) and to assess drug effects (Coull et al. 1996). Additionally some of these tests have been presented to marmosets (Roberts et al. 1988) to examine neuropsychological functioning. This comparative approach facilitates meaningful cross species comparison, particularly in the study of the effects of pharmacological intervention.

BACKGROUND: Lung transplant (LT) recipients requiring additional care may be referred to inpatient rehabilitation prior to discharge home. This study seeks to describe discharge destinations following LT, compare the characteristics of patients discharged to different destinations, and identify the predictors of discharge destination.METHODS: Retrospective study of 243 LT recipients who survived to hospital discharge between 2006 and 2009. LT recipients were compared based on discharge destination on data pertaining to demographics, clinical characteristics, and healthcare utilization.RESULTS: Of the 243 recipients, 197 (81%) were discharged home, 42 (17%) to inpatient rehabilitation, and 4 (2%) to other medical facilities. Age, pulmonary diagnosis, most recent six-minute walk distance (6 MWD) prior to transplant, pre-transplant mechanical ventilation, priority listing status, pre- and post-transplant intensive care unit length of stay (ICU LOS), post-transplant LOS, total LOS, and participation in pre-transplant rehabilitation were statistically different between patients that were discharged home versus inpatient rehabilitation. Age, most recent 6 MWD prior to transplant, pre-transplant mechanical ventilation, and total LOS were found to be independent predictors of discharge destination.CONCLUSION: Clinical factors can help identify patients more likely to require inpatient rehabilitation. Identification of these factors has the potential to facilitate early discharge planning and optimize continuity of care.

UNLABELLED: We propose a statistical model for estimating gene expression using data from multiple laser scans at different settings of hybridized microarrays. A functional regression model is used, based on a non-linear relationship with both additive and multiplicative error terms. The function is derived as the expected value of a pixel, given that values are censored at 65 535, the maximum detectable intensity for double precision scanning software. Maximum likelihood estimation based on a Cauchy distribution is used to fit the model, which is able to estimate gene expressions taking account of outliers and the systematic bias caused by signal censoring of highly expressed genes. We have applied the method to experimental data. Simulation studies suggest that the model can estimate the true gene expression with negligible bias.AVAILABILITY: FORTRAN 90 code for implementing the method can be obtained from the authors.

Introduction: Worldwide approximately 2.7 million are stillborn, more than 98% of these occur in developing countries. To address the problem, incidence and determinants of stillbirth must be understood. Therefore the aim of this study was to assess incidence and determinants of stillbirth among women who gave birth in Jimma University specialized hospital.Methods: A cross-sectional study design among 413 mothers who gave birth in Jimma specialized hospital was employed. Study subjects were selected by systematic sampling technique from the list of women who gave birth in hospital in one month study period. Data were collected by using pretested and structured questionnaire. Data were edited, cleaned, coded, entered and analyzed using SPSS-20 statistical software. Univarate and bivariate (logistic regressions) analysis was employed.Results: The incidence rate of stillbirth in the Hospital during a month period was 8% or 80 per 1000 total births. The predictors that showed an independent close association with stillbirth were absence of complication (OR = 0.1, 95% CI (0.04-0.2)), referral from other health facility (OR = 0.3, 95% CI (0.1-0.7)), having antenatal care (OR = 0.3, 95% CI (0.1-0.7)) and normal vaginal delivery (OR = 0.2, 95% CI ( 0.1-0.8)).Conclusion: The incidence rate of stillbirths in our setting is high and the identified determinants were related to both ante-partum and intra-partum-period. Therefore, effort should be made to improve antenatal, obstetric services and delivery services in terms awareness, access, timing and referral system to emergency care and specialized service to reduce the number of stillbirths.

OBJECTIVE: To analyze and compare external apical root resorption (EARR) of maxillary incisors treated by intrusion arch or continuous archwire mechanics.MATERIALS AND METHODS: This cone-beam computed tomography (CBCT) study analyzed 28 deep bite patients in the permanent dentition who were randomly divided into two groups: Group 1, 12 patients with initial mean age of 15.1 ± 1.6 years and mean overbite of 4.6 ± 1.2 mm treated with the Connecticut intrusion arch (CIA) in the upper arch (Ortho Organizers, Carlsbad, Calif) for a mean period of 5.8 ± 1.27 months. Group 2, 16 patients with initial mean age of 22.1 ± 5.7 years and mean overbite of 4.1 ± 1.1 mm treated with conventional leveling and alignment using continuous archwire mechanics for 6.1 ± 0.81 months. The degree of EARR was detected in 112 maxillary incisors by using CBCT scans and a three-dimensional program (Dolphin 11.7, Dolphin Imaging & Management Solutions, Chatsworth, Calif). The CBCT scans were obtained before (T1) and 6 months after initiation of treatment (T2). Differences between and within groups were assessed by nonpaired and paired t-tests, respectively, with a 5% significance level.RESULTS: Significant differences were found for both groups between T1 and T2 ( P < .05) indicating that EARR occurred in both groups. However, there were no significant differences when EARR was compared between group 1 (-0.76 mm) and group 2 (-0.59 mm).CONCLUSIONS: The Connecticut intrusion arch did not lead to greater EARR of maxillary incisors when compared with conventional orthodontic mechanics.

Radioimmunoassay was used to measure the blood content of beta-endorphines in patients with chronic alcoholism. The concentration of endogenous ethanol in these patients was determined by gas chromatography. The blood concentration of beta-endorphines was found to be high in patients who showed atypical affective disorders off the period of abstinence. It is assumed that peripheral beta-endorphine is not linked with the development of the narcomanic syndrome proper but mirrors the pathogenetic mechanisms of psychopathological disorders. The levels of endogenous ethanol vary in alcoholics and healthy subjects within the same ranges. However, the percentage distribution indicates that in patients, they are shifted toward lower concentrations, which is likely to be conditioned by the induction of enzymatic systems that metabolize ethanol.

Myrothecium verrucaria bilirubin oxidase expressed in Aspergillus oryzae is in a resting form different from that of the authentic bilirubin oxidase, but reaches the resting form of the authentic enzyme after one cycle of reduction and reoxidation with dioxygen as shown by the absorption and electron paramagnetic resonance spectra.

High-resolution photoelectron spectra of the gas phase pyrimidine-type nucleobases, thymine, uracil, and cytosine, were collected using synchrotron radiation over the photon energy range 17 ? hí ? 150 eV. These data provide the highest resolution photoelectron spectra of thymine, uracil, and cytosine published to date. By comparing integrated regions of the energy dependent photoelectron spectra of thymine, the ionization potentials of the first four ionic states of thymine were estimated to be 8.8, 9.8, 10.3, and 10.8 eV. The thymine data also show evidence for low energy shape resonances in three of the outermost valence electronic states. Comparing the uracil spectrum with the thymine spectrum, the four outermost valence electronic states of uracil likely begin at binding energies 9.3, 9.9, 10.5, and 11.0 eV. High-resolution spectra indicate only one tautomeric form of cytosine contributes significantly to the spectrum with the four outermost valence electronic states beginning at binding energies 8.9, 9.9, 10.4, and 10.85 eV.

This study explored the relationships among multimodal imaging, clinical features, and language impairment in patients with left temporal lobe epilepsy (LTLE). Fourteen patients with LTLE and 26 controls underwent structural MRI, functional MRI, diffusion tensor imaging, and neuropsychological language tasks. Laterality indices were calculated for each imaging modality and a principal component (PC) was derived from language measures. Correlations were performed among imaging measures, as well as to the language PC. In controls, better language performance was associated with stronger left-lateralized temporo-parietal and temporo-occipital activations. In LTLE, better language performance was associated with stronger right-lateralized inferior frontal, temporo-parietal, and temporo-occipital activations. These right-lateralized activations in LTLE were associated with right-lateralized arcuate fasciculus fractional anisotropy. These data suggest that interhemispheric language reorganization in LTLE is associated with alterations to perisylvian white matter. These concurrent structural and functional shifts from left to right may help to mitigate language impairment in LTLE.

Coadministration of anticancer drugs and multidrug resistance modulators directed against P-glycoprotein over-expressed in tumors also results in nonspecific blockade of this drug efflux pump in excretory tissues such as the liver and kidneys. These interactions often result in impaired renal and biliary clearance for anticancer agents such as doxorubicin (DOX). In the present investigation, we characterized the excretory processes associated with liposomal DOX administration to elucidate how liposome encapsulation may bypass adverse pharmacokinetic interactions between DOX and (3'-keto-Bmt1)-(Val2)-cyclosporin (Valspodar). Renal and biliary clearance properties of liposome-encapsulated DOX were compared with those for nonencapsulated DOX in the presence and absence of Valspodar using an instrumented rat model with implanted jugular vein and bile duct catheters for continuous sampling. Two types of liposomal DOX formulations were used, a drug-permeable egg phosphatidyl choline/cholesterol system and a sterically stabilized polyethylene glycol/1,2 distearoyl-sn-glycero-3-phosphocholine/cholesterol system to establish the relative roles of liposome-encapsulated and released drug on the pharmacokinetic and excretion alterations induced by Valspodar. DOX and its primary metabolites were quantitated using high-performance liquid chromatography. When Valspodar was coadministered with nonencapsulated DOX, 3.5- and 37.5-fold reductions in renal clearance (CLr) and biliary clearance (CLb), respectively, were observed, which resulted in increased plasma DOX concentrations and total exposure. However, Valspodar-induced alterations in CLr and CLb were less profound with egg phosphatidyl choline/cholesterol DOX (1.7- and 2.0-fold reductions, respectively) and negligible with the long-circulating polyethylene glycol-containing liposomal formulation. These results indicate that liposomes may circumvent Valspodar-induced DOX pharmacokinetic changes by reducing the rate of drug excretion in liver and kidney tissue to a level that is within the renal and biliary excretion capacity in the presence of P-glycoprotein blockade.

Haemodilution is always considerable during cardiopulmonary bypass (CPB). If this extra fluid sits in the muscle compartments then a corresponding rise in the compartment pressure (CP) is to be expected. The aim of this study was to measure pressure changes in a body compartment with new equipment, the MTC (Microtransducer). Changes in plasma colloid osmotic pressure (COP) were also measured during and after CPB to find a connection, if any, between CP and plasma COP. Ten elective consecutive CPB patients were studied. A 3-French (3-F) catheter-size electronic MTC was inserted in an anterior tibial compartment before CPB. The CP was monitored for 48 h. Plasma COP was also measured before, during and after CPB. CP increased significantly during and after CPB in all patients (p=0.01). COP decreased significantly in all patients (p=0.005), but no correlation was found between changes in COP and CP values in this study. Most of the patients reached their highest CP just after weaning off bypass. The CP remained elevated for 48 h, even though it then tended to decrease again. None of the patients reached the starting value within 48 h. COP decreased rapidly after going on bypass, but returned towards its starting value approximately 6 h after bypass. It is concluded that CP increases considerably during and after CPB and stays increased for at least 2 days after CPB. COP decreases during CPB, but reaches normal values 6 h after the CPB. No correlation was found between changes in CP and COP The MTC is a safe and easy way to measure intracompartment pressure.

Capabilities of methanol extracts from oregano and rosemary in retarding oxidation of long-chain polyunsaturated fatty acids, docosahexaenoic acid C22:6 (DHA) and eicosapentaenoic acid C20:5 (EPA), in menhaden oil were investigated. The fish oils after mixing with the extracts at different concentrations were oxidized in an accelerated study by heating at 150 degrees C for 30 min or incubating at 60 degrees C for 5 d. After heating at 150 degrees C, only 15.9% of DHA and 18.5% of EPA remained in the fish oil without extract, while 38.8% to 65.9% of DHA and 44.7% to 69.0% of EPA were retained in the fish oil mixed with 1% to 5% of oregano extract. The highest retained DHA (56.9%) and EPA (58.0%) in the fish oils mixed with rosemary extract were observed at 2.5% addition. Increasing rosemary extract to 5% lowered its capability of inhibiting DHA and EPA oxidation. After incubation at 60 degrees C for 5 d, the highest inhibition capability was also found at 2.5% of added rosemary extract, and the oil retained 88.2% DHA and 88.3% EPA. However, only 18.8% DHA and 23.6% EPA were retained in the fish oil mixed with 5% of oregano extract and no DHA and EPA were detected in the fish oil without extract after 5-d incubation at 60 degrees C. Thus, antioxidant activity of the rosemary extract was greater than that of oregano extract, but was sensitive to heat. The rosemary extract also demonstrated higher DPPH (2,2'-diphenyl-1-picrylhydrazyl) free radical scavenging capability, which was approximately 3 times higher than oregano extract, although there was no significant difference in the total phenolic contents between both extracts.

The accumulation of arsenic (As) in vegetables poses a risk of contamination to humans via the food chain. Two chard (var. cicla and var. d'ampuis) crops were grown for 60 days in greenhouses on Aridisol soil, and irrigated with water from Pastos Chicos, Jujuy (Argentina). The soil and water used in the trial presented 49 and 1.44 mg/L As concentration levels, respectively. Total dry biomass (TDB) and total As were determined in soils, roots and leaves. The latter was quantified by atomic absorption spectrometry with hydride generation, and bioconcentration and translocation factors were determined. TDB in var. cicla showed statistically significant differences when the plant was cultivated in control soil and watered with the toxicant (2.04 g), as compared with the treatment without exposure (2.8 g). TDB in var. d'ampuis presented statistically significant differences with respect to that of the control when the plants were grown in soils with As and watered with the toxicant (3.3 g). This variety increased its biomass in the presence of As. In the two Swiss chard varieties evaluated, the largest As accumulation in root and leaves was found when they were cultivated in contaminated soil and watered with distilled water. The presence of the toxicant in the leaves exceeded the limits established by C?digo Alimentario Argentino, i.e. 0.30 mg/kg. Total target hazard quotient (THQ) values for As were higher than 1, suggesting that consumers would run significant risks when consuming these chard varieties. Furthermore, it was determined that the carcinogenic risk (CR) posed by this type of exposure to As exceeded the acceptable risk level of 1 ? 10-6. Based on this evidence, we may conclude that consuming chard cultivated on the evaluated site brings about considerable risks to local residents' health.

Carcinoma arising in the apocrine sweat glands is rare and there are few reports describing the cytological features of this neoplasm. We describe the cytological features of a histologically confirmed apocrine carcinoma occurring in a 55-year-old man who presented with an ulcerated mass in the right axilla. Fine-needle aspiration cytology revealed features of a signet ring adenocarcinoma. The significance of this infrequently encountered neoplasm lies in its potential for diagnostic confusion with more common lesions containing signet ring cells. In an axillary mass lesion, cytological features along with clinical correlation are essential to distinguish primary apocrine carcinoma from mammary neoplasms with signet ring cells and other metastatic adenocarcinomas.

The authors report on a case of symptomatic Rathke's Cleft Cyst (RCC). The only clinical feature was headache. The cystic fluid was aspirated through a trans-sphenoidal approach. The postoperative course was uneventful. The rarity of symptomatic RCC (70 cases described in the literature; 30 operated patients) makes necessary the comparison of all data quoted in the different publications so as to define an appropriate attitude to treatment. It thus seems that the surgical procedure should be limited to an aspiration of the cyst contents through a trans-sphenoidal approach; such a procedure is more reliable than a craniotomy to avoid leakage of the cystic fluid into the subarachnoid space and possible postoperative aseptic meningitis. Some authors discussed the possibility of postoperative radiotherapy although only six patients developed recurrence of their clinical features several years after operation and also the efficacy of such a procedure has not been proved.

Desmoplastic fibroma (DF) is a rare bone tumor that accounts for about 0.1%-0.3% of all bone tumors. It is typically characterized as slow growing, but in rare cases it can proliferate extensively and exhibit locally aggressive characteristics. It is found most commonly in the appendicular skeleton and rarely in the axial skeleton. The authors present the cases of 2 women in their 20s with DF originating from the cervicothoracic spine. Both tumors intimately involved the brachial plexus and caused significant impingement of the mediastinum resulting in cardiopulmonary compromise. Both patients underwent hemiclamshell thoracotomies for tumor resection, and in both cases subtotal resection was performed given the encasement of the brachial plexus. Although DF is a benign process, it can be locally aggressive and proliferate at extensive rates. The authors describe these 2 cases, review the literature, and discuss management.

For producing a large quantity of human nerve growth factor beta (hNGF-beta), the recombinant adenovirus containing an expression cassette of hNGF-beta complementary DNA gene was constructed and then instilled directly into goats' mammary glands. The recombinant hNGF-beta was detected in the milk. The maximal expression level of recombinant hNGF-beta in the milk reached 196.8 mg/l. The biological activity of the recombinant hNGF-beta, compared with the commercial hNGF-beta, was tested using PC12 cells. The results demonstrate that instilling recombinant adenovirus directly into the mammary gland of goat is an efficient approach to producing a large quantity of hNGF-beta.

Major political and social changes in central and eastern European (CEE) countries offer new opportunities and challenges in tobacco control. Action against tobacco in CEE countries is part of an overall European strategy, the action plan for a tobacco-free Europe. Several CEE countries have voiced their concern about the deterioration of tobacco control and about the entry of the international tobacco industry. In response, the Regional Office has set up a framework for an international task force. The core of the task force consists of the EURO staff and a special consultant. Several European countries and organizations have already expressed their willingness to joint the task force by seconding an expert to participate in the country missions and consensus conferences. This paper describes particular steps to implement the action plan in central and eastern Europe. It outlines priority action to be carried out by the CEE countries themselves and describes possibilities for assistance by all European countries. A key role is assigned to a European alliance on tobacco or health to create psychological and material prerequisites for action as well as act in an advisory role. Some of the projects are already underway, but they can be considerably strengthened by extra resources and support. Some of the projects are possible only with new resources. The project proposals are classified according to the six areas of the new action plan but presented in a different order. This paper sets out a range of activities.(ABSTRACT TRUNCATED AT 250 WORDS)

Campylobacter jejuni and Campylobacter coli strains were isolated from feces of dairy cattle at farms with no known problem due to campylobacteria. Farms were located in the northeast, desert southwest, and Pacific west. Twenty isolates were identified by ribotyping with a RiboPrinter. The ability of these bovine isolates to colonize the ceca of chicks was determined by challenge inoculation and reisolation of the challenge strain from the ceca at 1 and 2 wk after challenge. Isolates recovered from chick ceca were examined by ribotyping to assure they matched the challenge strain. One hundred percent of the bovine-derived challenge strains were capable of colonizing chicks. These results indicate that dairy cattle may be asymptomatic Campylobacter carriers and potential sources of campylobacteria contamination of poultry facilities.

The field of robotic thoracic surgery has exploded in the last five years. Robotic technology allows the surgeon to perform complex operations with smaller incisions. As robotic systems become smaller, more efficient and the surgeons gain more experience, the results will continue to improve. The goal is less trauma to the patient which will decrease hospital stay, complications and lower health care costs, while allowing faster healing and productivity.

The role of neural cell adhesion molecule (NCAM) in the development and maturation of the neuromuscular junction (NMJ) was explored by characterizing structurally and functionally NMJs from postnatal day 11 (P11) to P30 +/+, +/-, and -/- NCAM null mutant mice. Differences in NCAM levels resulted in alterations in the size and shape of NMJs, with -/- NMJs being smaller. Additionally both the withdrawal of polyneuronal innervation and the selective accumulation of synaptic vesicle protein in the presynaptic terminal were delayed. These observations suggest that the bidirectional signaling responsible for these events is impaired at -/- NMJs. Functionally, miniature end plate potential size, end plate potential size, and quantal content did not differ from that of wild type under either normal or low release conditions. However at normal release conditions, -/- NMJs, unlike +/+ NMJs, lacked paired-pulse facilitation. The most striking abnormality was the inability of NCAM null junctions to maintain transmitter output with repetitive stimuli. Combined electrophysiological and FM1-43-labeling studies suggest that NCAM null junctions are unable either to dock or to mobilize a sufficient number of vesicles at high but physiological rates of transmitter release. Taken together our observations show that many aspects of transmission are normal and, thus, that many presynaptic and postsynaptic molecules have assembled properly in the absence of NCAM. However, the fact that NCAM was required for specific aspects of transmission, including paired-pulse facilitation and reliable transmission with repetitive stimuli, suggests that NCAM either is directly involved in these processes or is required for the proper organization and/or function of other molecules underlying these processes.

Despite reports of sex steroid receptor and COX2 expression in desmoid-type fibromatosis, responses to single agent therapy with anti-estrogens and non-steroidal anti-inflammatory drugs are unpredictable. Perhaps combination pharmacotherapy might be more effective in desmoid tumors that co-express these targets. Clearly, further understanding of the signaling pathways deregulated in desmoid tumors is essential for the development of targeted molecular therapy. Transforming growth factor-â (TGFâ) and bone morphogenetic proteins (BMP) are important regulators of fibroblast proliferation and matrix deposition, but little is known about the TGFâ superfamily in fibromatosis. A tissue microarray representing 27 desmoid tumors was constructed; 14 samples of healing scar and six samples of normal fibrous tissue were included for comparison. Expression of selected receptors and activated downstream transcription factors of TGFâ family signaling pathways, â-catenin, sex steroid hormone receptors and COX2 were assessed using immunohistochemistry; patterns of co-expression were explored via correlational statistical analyses. In addition to â-catenin, immunoreactivity for phosphorylated SMAD2/3 (indicative of active TGFâ signaling) and COX2 was significantly increased in desmoid tumors compared with healing scar and quiescent fibrous tissue. Low levels of phosphorylated SMAD1/5/8 were detected in only a minority of cases. Transforming growth factor-â receptor type 1 and androgen receptor were expressed in both desmoid tumors and scar, but not in fibrous tissue. Estrogen receptor-â was present in all cases studied. Transforming growth factor-â signaling appears to be activated in desmoid-type fibromatosis and phosphorylated SMAD2/3 and COX2 immunoreactivity might be of diagnostic utility in these tumors. Given the frequency of androgen receptor, estrogen receptor-â and COX2 co-expression in desmoid tumors, further assessment of the efficacy of combination pharmacotherapy using hormonal agonists/antagonists together with COX2 inhibitors should be considered.

PURPOSE: To study selective use of antibiotic-impregnated catheters in children at increased risk of venous catheter-related infections (CRIs).MATERIALS AND METHODS: From December 2008 to June 2009, 428 peripherally inserted central catheters (PICCs) were placed by the interventional radiology service of a large metropolitan children's hospital. This retrospective study analyzed demographic and outcome data for the 125 patients in this group at high risk for venous CRI. Patients at high risk were those with active systemic infection, previous complicated central venous access, intensive care unit (ICU) admission, intestinal failure, transplantation, complex congenital heart disease, or renal failure. Patients (age, 7.6 y ± 7.0; 73 male and 52 female) received a conventional or antibiotic-impregnated PICC, with 17 receiving more than one catheter.RESULTS: Of the 146 of 428 qualifying patient encounters (34%), 53 patients received an antibiotic-impregnated PICC and 93 received a conventional PICC, representing 5,080 total catheter-days (CDs). The rates of CRIs per 1,000 CDs, including catheter exit site infections and catheter-related bloodstream infections, were 0.86 for antibiotic-impregnated PICCs and 5.5 for conventional PICCs (P = .036). A propensity-based model predicts 15-fold greater infection-free survival over the lifetime of the catheter in patients who receive an antibiotic-impregnated PICC (P < .001). Antibiotic-impregnated PICC recipients with active infection or ICU admission at the time of insertion had no catheter-associated infections, compared with 3.42 and 9.46 infections per 1,000 CDs, respectively, for patients who received conventional PICCs. Patients with intestinal failure had 1.49 and 10 infections per 1,000 CDs with antibiotic-impregnated versus conventional PICCs, respectively.CONCLUSIONS: Antibiotic-impregnated long-term PICCs significantly improve infection-free catheter survival in pediatric patients at high risk.

SETTING: The network of supranational tuberculosis reference laboratories (SRLs).OBJECTIVE: To evaluate the annual SRL Rounds 6-14 of proficiency testing for first-line drug susceptibility testing (DST).DESIGN: Panels consisted of 20-30 cultures (including 10 pairs of duplicate strains), aiming at 50% resistance prevalence with a variety of profiles. The 27 SRLs participating in at least one of these rounds were free to use their preferred DST method. A judicial gold standard of at least 80% concordant 'susceptible' or 'resistant' was used to determine sensitivity, specificity and efficiency; otherwise the strain was excluded.RESULTS: Of 600 strains, 10% were excluded from evaluation. The average SRL sensitivity and specificity varied between rounds, without attaining significance or trends. Both sensitivity and specificity remained at >95% for isoniazid (INH), rifampicin (RMP) and streptomycin and at >80% for ethambutol. The 16 SRLs participating in all rounds performed consistently better.CONCLUSION: The rounds succeeded in comparing the proficiency of laboratories, and should be further promoted for DST quality assessment. However, to function with greater precision and to ultimately improve the clinical relevance of DST, the INH and RMP judicial result gold standard also needs to take into account genotypic and treatment outcome information.

The Chinese mitten crab Eriocheir sinensis is an economically important aquatic species in China. Many studies on gene structure, breeding, and diseases of the crab have been reported. However, knowledge about the organization of the nerve system of the crab remains largely unknown. To study the ultrastructure of the cerebral ganglia of E. sinensis and to compare the histological findings regarding the nerve systems of crustaceans, the cerebral ganglia were observed by transmission electron microscopy. The results showed that four types of gliocytes, including type I, II, III, and IV gliocytes were located in the cerebral ganglia. In addition, three types of synapses were present in the cerebral ganglia, including unidirectional synapses, bidirectional synapses, and combined type synapses.

Dietary restriction is one of the several ways which could putatively extend organisms' lifespan, ranging from Saccharomyces cerevisiae to rodents, by activating the AMP-activated protein kinase (AMPK), an ATP/AMP sensor. Extensive researches have shown that aging reduces sensibility of AMPK and eventually causes energy imbalance in cells. Research in mammals' AMPK depicts that this signaling molecule could control autophagy, improve cellular stress resistance and suppress inflammatory responses. Hence, in this study we performed a drug repurposing of 1908 FDA-approved drugs in order to discover putative safe activators of AMPK and to find new applications for existing drugs. For this purpose, FDA-approved drugs were screened by virtual screening and the ligand-protein interactions were carefully inspected. Moreover, through MM/PBSA analysis, the binding affinity of hit compounds in ã and áâ binding sites were investigated. As Cangrelor, Nacitentan, Levoleucovorin and Glisoxepide had lower binding affinities; we predicted that they would probably prove to be more potential activators than C2. However, hit-compounds in áâ binding site, exhibited higher unfavorable binding affinity. Hence, present findings can prove to be valuable for discovering new activators for AMPK.

The steady-state mechanism of the aminoacylation of tRNAPhe by the corresponding synthetase from yeast has been investigated in detail by kinetic experiments. It was found that there are two alternative mechanisms: one favoured at low tRNA concentrations and the other at high tRNA concentrations. ATP and Phe are bound randomly to the enzyme. AMP is released immediately after the binding of ATP and Phe. Between the release of AMP and pyrophosphate (PPi) there is at least one additional step. Based on the experimental results a model of the steady-state mechanism is proposed. This model includes the sequence of addition of substrates to the enzyme and the release of products from the enzyme as well as the composition of the intermediate complexes with the enzyme. This model is in accordance with previous results based on different techniques. The results are explained by a "flip-flop" mechanism for all the substrates and products involved in the reaction.

BACKGROUND: Chronic suppurative otitis media can be recalcitrant and difficult to treat, particularly with the increasing occurrence of antibiotic resistance. Lactobacillus plantarum is a probiotic that has been shown to decrease S. aureus and P. aeruginosa growth in wounds, making it a good candidate for the treatment of chronic suppurative otitis media. However, before it can be applied in the ear, its ototoxicity potential must be evaluated.METHODS: A prospective controlled trial was conducted in a chinchilla animal model at the Animal care research facilities of the Montreal Children's Hospital Research Institute to determine whether Lactobacillus plantarum is ototoxic when applied transtympanically. Ten chinchillas each had one ear randomly assigned to receive 109 CFU/mL of Lactobacillus plantarum solution, while the contralateral ear received saline. Auditory brainstem responses were measured bilaterally at 8, 20, 25 kHz before, at 7-10 days after application, and at 28 days after application of probiotic or saline. Facial nerve and vestibular function were assessed clinically.RESULTS: There were no statistically significant differences in hearing thresholds between control and experimental ears at 28 days after application. A difference of 11 dB was noted in the 25 kHz range at day 7-10, but resolved by day 28. No animals receiving probiotics developed vestibular nerve dysfunction. There was no histologic evidence of auditory hair cell damaged evidenced by scanning electron microscopy.CONCLUSION: Our study suggests that a single application of Lactobacillus plantarum at 109 CFU/mL does not cause ototoxicity in a chinchilla animal model. These preliminary safety evaluations and the pathogen inhibitory effects of L. plantarum demonstrated by previous studies present this probiotic as a candidate of interest for further investigation.

Cultured human retinal pigmented epithelial cells were studied using phosphorus-31 nuclear magnetic resonance spectroscopy (P-31 NMR). Retinal pigmented epithelial cells from normal human donors were isolated and expanded using roller-bottle culture. The P-31 NMR spectrum of the intact living cells was obtained at 121 MHz by casting the cells in agarose threads and perfusing the threads with culture medium. The cells remained viable at 37 degrees C in the NMR magnet for at least 24 hr, as determined by the stability of the phosphorus spectrum and by trypan blue dye exclusion at the end of the experiments. The intact cell spectrum showed resonances from phosphorylethanolamine, phosphorylcholine, inorganic phosphate, phosphodiesters, phosphocreatine, ATP, NAD+, and UDP-N-acetylglucosamine, with phosphorylethanolamine, phosphorylcholine, and ATP being the major metabolites present. The resonances were assigned by making a perchloric acid (PCA) extract of the intact cells and running under high-resolution conditions. The PCA extract spectrum also detected sugar phosphates, ADP and UTP, with the latter being approximately 20% of the nucleotide pool. These studies provide the basis for the study of normal and diseased human RPE cells by NMR spectroscopic methods.

BACKGROUND: Despite the significant access and cost-saving potential of telehealth, the uptake of telehealth services in Australia has been sporadic. Understanding the factors that drive the uptake of home-telehealth services from the consumer perspective has received scant attention in the literature.AIM: The aim of this study was to explore how a comprehensive set of factors may influence the intention of older Australians to adopt home telehealth services.METHODS: A survey of 306 Australians aged between 50 and 68 years was conducted to examine the influence of six categories of predictors on the intention of older Australians to adopt home telehealth: (a) demographics, (b) health status and usage, (c) mobility and ease of access to healthcare, (d) technology usage and anxiety with technology, (e) telehealth attitudes, and (f) personality traits.RESULTS: Hierarchical regression analysis revealed that significant predictors were: trust in telehealth (â = 0.35); the technology acceptance model (â = 0.27); healthcare habits (â = -0.20); dissatisfaction with traditional healthcare (â = 0.19) and online behaviors (â = 0.09). The model explained 63% of the variance in intention to adopt home telehealth.CONCLUSION: This study is the first of its kind in Australia and provides valuable insight into the factors which impact consumer's intention to adopt telehealth services.

How should robotic or prosthetic arms be programmed to move? Copying human smooth movements is popular in synthetic systems, but what does this really achieve? We cannot address these biomimetic issues without a deep understanding of why natural movements are so stereotyped. In this article, we distinguish between 'functional' and 'aesthetic' biomimetics. Functional biomimetics requires insight into the problem that nature has solved and recognition that a similar problem exists in the synthetic system. In aesthetic biomimetics, nature is copied for its own sake and no insight is needed. We examine the popular minimum jerk (MJ) model that has often been used to generate smooth human-like point-to-point movements in synthetic arms. The MJ model was originally justified as maximizing 'smoothness'; however, it is also the limiting optimal trajectory for a wide range of cost functions for brief movements, including the minimum variance (MV) model, where smoothness is a by-product of optimizing the speed-accuracy trade-off imposed by proportional noise (PN: signal-dependent noise with the standard deviation proportional to mean). PN is unlikely to be dominant in synthetic systems, and the control objectives of natural movements (speed and accuracy) would not be optimized in synthetic systems by human-like movements. Thus, employing MJ or MV controllers in robotic arms is just aesthetic biomimetics. For prosthetic arms, the goal is aesthetic by definition, but it is still crucial to recognize that MV trajectories and PN are deeply embedded in the human motor system. Thus, PN arises at the neural level, as a recruitment strategy of motor units and probably optimizes motor neuron noise. Human reaching is under continuous adaptive control. For prosthetic devices that do not have this natural architecture, natural plasticity would drive the system towards unnatural movements. We propose that a truly neuromorphic system with parallel force generators (muscle fibres) and noisy drivers (motor neurons) would permit plasticity to adapt the control of a prosthetic limb towards human-like movement.

This study compared the repertoire of cytokines produced by epithelial cell lines and human peripheral blood monocytes in response to Escherichia coli. The A-498 and J82 urinary tract epithelial cell lines and human peripheral blood monocytes were exposed to E. coli Hu734. The cytokine content of single cells was detected by indirect immunofluorescence using monoclonal antibodies to interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF-alpha), TNF-beta, IL-6, IL-8, and granulocyte macrophage-colony-stimulating factor, and the number of positive cells was used to quantitate the response. The J82 bladder cell line stained positive for IL-6, IL-8, and IL-1 alpha. The IL-8 and IL-6 response peaked at 2 h, while the number of IL-1 alpha-positive cells reached a peak 6 h after E. coli stimulation. The A-498 kidney cell line stained for IL-8 with a peak at 2 h and IL-6 with a peak at 6 h after E. coli stimulation. Peripheral blood monocytes stained for the cytokines IL-1 alpha, IL-1 beta, IL-8, IL-6, and TNF-alpha but not for TNF-beta and granulocyte macrophage-colony-stimulating factor after stimulation with E. coli. The results demonstrated that bacteria activated a cytokine response in the epithelial cell lines and monocytes. The epithelial cell lines had a more limited cytokine response profile than circulating monocytes, which may serve to limit the consequences of microbial exposure at the mucosal surface and help maintain the integrity of other tissue compartments.

BACKGROUND: The Multidimensional Prognostic Index (MPI) has been found to predict mortality in patients with a variety of clinical conditions. We aimed to assess the association of the MPI with future mortality and number of in-hospital days for the first time in a population-based cohort.METHODS: The study population consisted of 2472 persons, aged 66-99 years, from the Swedish National Study on Aging and Care in Kungsholmen, Sweden, who underwent the baseline visit 2001-4, and were followed up >10 years for in-hospital days and >12 years for mortality. The MPI was a modified version of the original and aggregated seven domains (personal and instrumental activities of daily living, cognitive function, illness severity and comorbidity, number of medications, co-habitation status, and nutritional status). The MPI score was divided into risk groups: low, medium and high. Number of in-hospital days (within 1, 3 and 10 years) and mortality data were derived from official registries. All analyses were age-stratified (sexagenarians, septuagenarians, octogenarians, nonagenarians).RESULTS: During the follow-up 1331 persons (53.8%) died. Laplace regression models, suggested that median survival in medium risk groups varied by age from 2.2-3.6 years earlier than for those in the corresponding low risk groups (p = 0.002-p<0.001), and median survival in high risk groups varied by age from 3.8-9.0 years earlier than for corresponding low risk groups (p<0.001). For nonagenarians, the median age at death was 3.8 years earlier in the high risk group than for the low risk group (p<0.001). The mean number of in-hospital days increased significantly with higher MPI risk score within 1 and 3 years for people of each age group.CONCLUSION: For the first time, the effectiveness of MPI has been verified in a population-based cohort. Higher MPI risk scores associated with more days in hospital and with fewer years of survival, across a broad and stratified age range.

In Friedreich's ataxia (FRDA) patients, diminished frataxin (FXN) in sensory neurons is thought to yield the predominant pathology associated with disease. In this study, we demonstrate successful usage of RNA transcript therapy (RTT) as an exogenous human FXN supplementation strategy in vitro and in vivo, specifically to dorsal root ganglia (DRG). Initially, 293 T cells were transfected with codon optimized human FXN mRNA, which was translated to yield FXN protein. Importantly, FXN was rapidly processed into the mature functional form of FXN (mFXN). Next, FXN mRNA, in the form of lipid nanoparticles (LNPs), was administered intravenously in adult mice. Examination of liver homogenates demonstrated efficient FXN LNP uptake in hepatocytes and revealed that the mitochondrial maturation machinery had efficiently processed all FXN protein to mFXN in ~24 h in vivo. Remarkably, greater than 50% mFXN protein derived from LNPs was detected seven days after intravenous administration of FXN LNPs, suggesting that the half-life of mFXN in vivo exceeds one week. Moreover, when FXN LNPs were delivered by intrathecal administration, we detected recombinant human FXN protein in DRG. These observations provide the first demonstration that RTT can be used for the delivery of therapeutic mRNA to DRG.

We describe here a simple procedure, by which HLA class II antigens can be accurately and reliably identified in those patients where there is minimal or absent expression of HLA-DR,DQw antigens on B cells, or when the total number of leukocytes recovered from the patients do not permit reliable typing. Ficoll-Hypaque-separated peripheral blood mononuclear leukocytes, fresh or cryopreserved, were activated by PHA and then propagated in IL-2-containing medium until enough cells for typing were obtained (usually 7-14 days). At this stage, the cultured cells were shown to be primarily T cells (greater than 90% CD3+). Since the activated T cells propagate in the presence of IL-2, even a small number (10(4] of fresh or cryopreserved patients' cells suffice for this protocol. To date we have been able to successfully HLA-DR,DQw type 34/34 bone marrow transplantation candidates and 12/12 long-term dialysis patients, who were untypable using fresh cells. HLA-DR,DQw antigens on activated T cells from normal individuals were identical to those found on their uncultured B cells. In addition, class I antigens that were undetectable on the uncultured cells of one patient could be identified on activated T cells. The HLA antigens identified on the patients' activated T cells were confirmed by phenotypic analysis of cells from family members.

The success of the Ilizarov technique is due to the combination of the biomechanics of its external fixator and the biology of distraction osteogenesis. The stability and stiffness of the conventional Ilizarov fixator is attributed to its use of a K-wire cross structure. The disadvantages of this structure include pain, and possible neurologic and vascular injuries when the wires are introduced into crucial neurovascular areas, as well as increased frame complexity and construction. Reducing the number of wires decreases these problems, but also decreases the stiffness of the system. Hybrid (wire and half-pins) Ilizarov fixators and half-pin fixators are also being used in an attempt to alleviate these problems. Half-pins have been described by Fleming et al. and Green as causing minimal transfixation of the surrounding soft tissues and capable of and being inserted into anatomically safe areas. The stiffnesses of the hybrid systems have not been measured. This study reports on the stiffness of different wire and half-pin systems that were biomechanically tested in axial loading, anterior-posterior bending, medial-lateral bending, and torsional loading. The results demonstrated that the conventional Ilizarov fixator with wires possesses a high axial stiffness, whereas the fixator with half-pins possesses higher stiffness under bending and torsional loads. To obtain adequate stability, the use of hybrid Ilizarov frames with one wire and two or more half-pins (5 or 6 mm in diameter), or larger half-pin frames (5 or 6 mm) with three pins is recommended.

BACKGROUND: Most studies on sacral nerve stimulation (SNS) are either single-centre with small numbers of patients or multi-centre studies. We present the medium-term follow-up results from a single centre for 127 patients undergoing SNS.METHODS: Consecutive patients treated with SNS for faecal incontinence had preoperative baseline St Mark's continence scores, faecal incontinence quality of life (FIQL) measures and anorectal physiology studies. Follow-up was a postal questionnaire concerning continence, FIQL, patient-perceived change in bowel control (-5 to +5 where 0 is no change), overall satisfaction (0-10 visual analogue scale) and use of medications.RESULTS: A total of 166 patients underwent temporary nerve stimulation testing, of which 112 progressed to a permanent implantable pulse generator (IPG). Fifteen received an IPG without the testing phase, hence 127 patients in total. Fourteen had the IPG removed, four were deceased, leaving 109 for assessment; 91 (83%) responded to the survey. Mean follow-up was 2.7 years (range: 2 months-8.5 years). Mean baseline St Mark's continence score was 14.4, and mean follow-up score was 10.3 (P < 0.01). FIQL improved in all domains (P < 0.001). Patient-reported improved bowel control mean score was +3.2 (95% CI: 2.9, 3.55). Median satisfaction score was 8.0 (range: 0-10). Complications included 17 lead dislodgements, seven superficial infections, five infections requiring surgery and five repositioning of a rotated IPG. Thirty-two patients used loperamide and 34 used fibre supplements.CONCLUSION: In this observational study, limited by the absence of a placebo control group, SNS significantly improved continence and quality of life, and patient satisfaction was high.

The decline in the number of children nationally being vaccinated against pertussis has caused anxiety and concern, both for parents and for professional workers. Publicity by the media and professional press has added to the confusion, and has been largely held responsible for the present situation. A survey was undertaken to analyse the factors which may have been responsible for the reduction in the number of children coming forward for pertussis vaccination in Leicestershire. These factors included social class, the position of the child within the family, and the reasons given by families for omission of the vaccine. Observations on the computerized record of immunization and vaccination, and the family's own record of the dates of immunization were made. The results of the survey present a more complicated picture than expected, with the consequence that solutions are difficult to find.