Determination and prediction of the magnetic anisotropy of Mn ions.
This tutorial is dedicated to the investigation of magnetic anisotropy using both electron paramagnetic resonance (EPR) spectroscopy for its experimental determination and quantum chemistry for its theoretical prediction. Such an approach could lead to the definition of magneto-structural correlation essential for the rational design of complexes with targeted magnetic properties or for the identification of unknown reactive metallic species involved in catalysis. To illustrate this combined approach the high spin MnII, MnIII and MnIV ions have been taken as specific examples. The first part deals with the analysis of the EPR experiments as a function of the ions under investigation and the conditions of the measurements, specifically: (i) EPR spectra recorded under high vs. low frequency conditions with respect to magnetic anisotropy, (ii) EPR spectra of non-integer (Kramers) vs. integer (non-Kramers) spin states and (iii) mono- vs. multi-frequency EPR spectra. In the second part, two main quantum chemical approaches, which have proven their capability to predict magnetic anisotropy, are described. More importantly, these calculations give access to the different contributions of zero field splitting, key information for the full understanding of magnetic anisotropy. The last part demonstrates that such a combined experimental and theoretical approach allows for the definition of magneto-structural correlations.


Multiresolution infrared optical properties for Gaussian sea surfaces: theoretical validation in the one-dimensional case.
The validation of the multiresolution model of sea surface infrared optical properties developed at ONERA is investigated in the one-dimensional case by comparison with a reference model, using a submillimeter discretization of the surface. Having expressed the optical properties, we detail the characteristics of each model. A set of numerical tests is made for various wind speeds, resolutions, and realizations of the sea surface. The tests show a good agreement between the results except for grazing angles, where the impact of multiple reflections and the effects of adjacent rough surfaces on shadow have to be investigated.


Anticonvulsant Effects of Fractions Isolated from Dinoponera quadriceps (Kempt) Ant Venom (Formicidae: Ponerinae).
Natural products, sources of new pharmacological substances, have large chemical diversity and architectural complexity. In this context, some toxins obtained from invertebrate venoms have anticonvulsant effects. Epilepsy is a neurological disorder that affects about 65 million people worldwide, and approximately 30% of cases are resistant to pharmacological treatment. Previous studies from our group show that the denatured venom of the ant Dinoponera quadriceps (Kempt) protects mice against bicuculline (BIC)-induced seizures and death. The aim of this study was to investigate the anticonvulsant activity of compounds isolated from D. quadriceps venom against seizures induced by BIC in mice. Crude venom was fractionated by high-performance liquid chromatography (HPLC) resulting in six fractions referred to as DqTx1-DqTx6. A liquid chromatography-mass spectrometry (LC/MS) analysis revealed a major 431 Da compound in fractions DqTx1 and DqTx2. Fractions DqTx3 and DqTx4 showed a compound of 2451 Da and DqTx5 revealed a 2436 Da compound. Furthermore, the DqTx6 fraction exhibited a major component with a molecular weight of 13,196 Da. Each fraction (1 mg/mL) was microinjected into the lateral ventricle of mice, and the animals were observed in an open field. We did not observe behavioral alterations when the fractions were given alone. Conversely, when the fractions were microinjected 20 min prior to the administration of BIC (21.6 nM), DqTx1, DqTx4, and DqTx6 fractions increased the latency for onset of tonic-clonic seizures. Moreover, all fractions, except DqTx5, increased latency to death. The more relevant result was obtained with the DqTx6 fraction, which protected 62.5% of the animals against tonic-clonic seizures. Furthermore, this fraction protected 100% of the animals from seizure episodes followed by death. Taken together, these findings indicate that compounds from ant venom might be a potential source of new anticonvulsants molecules.


A simple method of equine limb force vector analysis and its potential applications.
Ground reaction forces (GRF) measured during equine gait analysis are typically evaluated by analyzing discrete values obtained from continuous force-time data for the vertical, longitudinal and transverse GRF components. This paper describes a simple, temporo-spatial method of displaying and analyzing sagittal plane GRF vectors. In addition, the application of statistical parametric mapping (SPM) is introduced to analyse differences between contra-lateral fore and hindlimb force-time curves throughout the stance phase. The overall aim of the study was to demonstrate alternative methods of evaluating functional (a)symmetry within horses. GRF and kinematic data were collected from 10 horses trotting over a series of four force plates (120 Hz). The kinematic data were used to determine clean hoof contacts. The stance phase of each hoof was determined using a 50 N threshold. Vertical and longitudinal GRF for each stance phase were plotted both as force-time curves and as force vector diagrams in which vectors originating at the centre of pressure on the force plate were drawn at intervals of 8.3 ms for the duration of stance. Visual evaluation was facilitated by overlay of the vector diagrams for different limbs. Summary vectors representing the magnitude (VecMag) and direction (VecAng) of the mean force over the entire stance phase were superimposed on the force vector diagram. Typical measurements extracted from the force-time curves (peak forces, impulses) were compared with VecMag and VecAng using partial correlation (controlling for speed). Paired samples t-tests (left v. right diagonal pair comparison and high v. low vertical force diagonal pair comparison) were performed on discrete and vector variables using traditional methods and Hotelling's T2 tests on normalized stance phase data using SPM. Evidence from traditional statistical tests suggested that VecMag is more influenced by the vertical force and impulse, whereas VecAng is more influenced by the longitudinal force and impulse. When used to evaluate mean data from the group of ten sound horses, SPM did not identify differences between the left and right contralateral limb pairs or between limb pairs classified according to directional asymmetry. When evaluating a single horse, three periods were identified during which differences in the forces between the left and right forelimbs exceeded the critical threshold (p < .01). Traditional statistical analysis of 2D GRF peak values, summary vector variables and visual evaluation of force vector diagrams gave harmonious results and both methods identified the same inter-limb asymmetries. As alpha was more tightly controlled using SPM, significance was only found in the individual horse although T2 plots followed the same trends as discrete analysis for the group. The techniques of force vector analysis and SPM hold promise for investigations of sidedness and asymmetry in horses.


Mycobacterium paraintracellulare sp. nov., for the genotype INT-1 of Mycobacterium intracellulare.
Three mycobacterial strains, isolated from independent Korean patients with pulmonary infections, belonging to the Mycobacterium intracellulare genotype 1 (INT-1) were characterized using a polyphasic approach. The sequences of the 16S rRNA gene and internal transcribed spacer 1 (ITS1) of the INT-1 strains were identical to those of Mycobacterium intracellulare ATCC 13950T. However, multilocus sequence typing (MLST) analysis targeting five housekeeping genes (hsp65, rpoB, argG, gnd and pgm) revealed the phylogenetic separation of these strains from M. intracellulare ATCC 13950T. DNA-DNA hybridization values of >70 % confirmed that the three isolates belong to the same species, while the values of <70 % between one of them and the type strains of M. intracellulare and Mycobacterium chimaera confirmed their belonging to a distinct species. In addition, phenotypic characteristics such as positive growth on MacConkey agar and in acidic broth culture, unique matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS profiles of lipids, and unique mycolic acids profiles further supported the taxonomic status of these strains as representatives of a novel species of the Mycobacterium avium complex named Mycobacterium paraintracellulare. The type strain is MOTT64T (=KCTC 29084T=JCM 30622T).


Surgical resection versus laparoscopic radiofrequency ablation in patients with hepatocellular carcinoma and Child-Pugh class a liver cirrhosis.
This study compared two homogeneous groups of patients submitted to either surgical resection (HR) or laparoscopic radiofrequency ablation (LRFA) for the treatment of hepatocellular carcinoma (HCC). When compatible with the liver functional reserve, HR remains the treatment of choice for HCC, while LRFA seems to be a promising, less invasive alternative. We thus compared HR or LRFA for short- and long-term outcomes in patients with a single HCC nodule and Child-Pugh class A liver cirrhosis. We enrolled 152 cirrhotic patients out of 372 cases consecutively evaluated for HCC. Enrolled patients with similar baseline characteristics underwent HR (n = 78) or LRFA (n = 74), in both cases with intraoperative ultrasonography, and they were then followed for similar durations (mean +/- standard deviation, 36.2 +/- 23.5 months for HR vs. 38.2 +/- 28.4 for LRFA). Outcomes included short- and long-term morbidity, HCC recurrence, and overall survival. Short-term morbidity was far higher in the HR group while, during follow-up, HCC recurrence (mainly local) was more frequent in patients treated with LRFA. More importantly, baseline alfa-fetoprotein levels and early HCC recurrence after treatment greatly influenced overall survival, while the use of HR or LRFA did not predict it. On the other hand, HCC recurrence was found to be determined by the surgical approach and ultrasound characteristics of the tumor. Our data were obtained from a large number of HCC cases and support similar survival rates after HR or LRFA for single HCC nodules on Child-Pugh class A liver cirrhosis, despite a marked increase in HCC recurrence rates after LRFA.


Disentangling the effects of circulating IGF-1, glucose, and cortisol on features of perceived age.
Circulatory levels of insulin-like growth factor (IGF-1), glucose, and cortisol have been previously associated with facial aging. However, as these serum measures are related, it is unclear whether their associations with skin aging occur independently from each other. We aimed to investigate whether the associations between serum IGF-1, glucose, and cortisol levels and perceived age/wrinkle grade occur independently of each other and whether these are mediated via skin wrinkling or via other skin aging features. Perceived age and skin wrinkling grade were assessed in a random sample from the Leiden Longevity Study with non-fasted (N = 579) and fasted blood sampling (N = 219). In our study population, a higher non-fasted IGF-1 level was associated with a lower skin wrinkling grade (p value = 0.014) and tended to associate with a lower perceived age (p value = 0.067), which was mediated for approximately 100 % by skin wrinkling. A higher non-fasted glucose level was associated with a higher perceived age (p value = 0.017), which was mediated for 51 % by skin wrinkling grade (p value = 0.112). A higher fasted cortisol level tended to associate with a higher perceived age (p value = 0.116), which was mediated for 29 % by skin wrinkling. Results remained similar when the serum measures were statistically adjusted for each other. Thus, the previously reported serum measures associate independently from each other with skin aging. IGF-1 is predominantly associated with perceived age by skin wrinkling, whereas cortisol and glucose also by other skin aging features.


Risk factors for supplementary posterior instrumentation after anterolateral decompression and instrumentation in thoracolumbar burst fractures.
In spite of the established benefits of anterolateral decompression and instrumentation (ALDI) for thoracolumbar burst fractures (TLBF), the indications for supplementary posterior instrumentation remain unclear. A retrospective review of clinical and radiographic data of a prospective cohort of 73 patients who underwent ALDI for TLBF from T12 to L4. The mean age of the cohort was 42 ± 15 years, with 49 males and 24 females. Forty-six patients had neurological deficit, and 27 were intact. Owing to symptomatic settling, supplemental posterior instrumentation was performed in 7 out of 73 patients. The age of patients requiring supplemental posterior instrumentation (59 ± 14 years) exceeded that of patients who did not (41 ± 16, p=0.004). Otherwise, the patients who required posterior instrumentation were comparable to those treated with ALDI in terms of body mass index (BMI), American Spinal Injury Association (ASIA) scores on admission and follow-up, residual spinal canal, and local kyphosis on admission and follow-up. The posterior ligamentous complex (PLC) integrity was assessed in 38 patients in whom the MRI scans were retrievable, 31 successfully treated with ALDI, and all 7 undergoing supplementary posterior instrumentation. Subgroup analysis demonstrated that there was no difference in the incidence of PLC disruption between the 2 groups (p=0.257). Secondary supplemental posterior instrumentation was deemed necessary in 10% of cases following ALDI. Age was the only significant risk factor predicating supplemental posterior instrumentation.


Down-regulation of NLRP3 inflammasome in gingival fibroblasts by subgingival biofilms: involvement of Porphyromonas gingivalis.
Recognition of pathogen-associated molecular patterns that activate IL-1β is regulated by inflammasomes, predominantly of the nucleotide-binding oligomerization domain-like receptor (NLR) family. NLRP3 inflammasome is involved in the innate immune responses in periodontal disease. This is an inflammatory condition that destroys the tooth-supporting (periodontal) tissues, initiated by the subgingival formation of multi-species biofilms, frequently including the Gram-negative species Porphyromonas gingivalis. The aim of this study was to investigate the relative effect of P. gingivalis as part of subgingival biofilm, on the expressions of NLRP3 inflammasome, absent in melanoma (AIM)2 (a non-NLR inflammsome) and IL-1β by human gingival fibroblasts. The 10-species subgingival biofilm model, or its 9-species variant excluding P. gingivalis, were used to challenge the cells for 6 h. Gene expression analysis for various inflammasome components and IL-1β was performed by TaqMan real-time PCR. The 10-species subgingival biofilm reduced NLRP3 and IL-1β, but did not affect AIM2 expression. Exclusion of P. gingivalis from the biofilm partially rescued NLRP3 and IL-1β expressions. In conclusion, subgingival biofilms down-regulate NLRP3 and IL-1β expression, partly because of P. gingivalis. These dampened host innate immune responses may favour the survival and persistence of the associated biofilm species in the periodontal tissues.


Intracytoplasmic delivery of listeriolysin O by a vaccinal strain of Bacillus anthracis induces CD8-mediated protection against Listeria monocytogenes.
The facultative intracellular pathogen Listeria monocytogenes secretes a 58-kDa hemolysin, listeriolysin O (LLO), that allows bacteria to access the cytoplasm and to multiply inside infected cells. LLO is also a protective Ag required for the development of specific immunity. We studied the capacity of a new bacterial vector, derived from an attenuated strain of Bacillus anthracis, to deliver in vivo LLO and to induce protection against L. monocytogenes infection. The hly gene encoding LLO was fused to a B. anthracis regulatory region induced in vivo and was integrated into a resident plasmid of this bacterium. This recombinant strain secreted a functional LLO in vitro and inside phagosomes of bone marrow macrophages. This LLO production enabled the conversion of the extracellular replicating B. anthracis into an intracytoplasmic bacterium. LLO+ B. anthracis thus mimicked the intracellular behavior of L. monocytogenes in macrophages. Specific protection of mice against lethal doses of L. monocytogenes was induced by immunization with LLO+ B. anthracis. The immunity was mediated by CD8+ T lymphocytes and was associated with the activation of LLO-specific MHC class I-restricted CD8+ CTL, able to recognize the immunodominant H-2d-restricted epitope 91-99 of LLO. This study, therefore, suggests that intracytoplasmic delivery of LLO by B. anthracis is sufficient to induce a MHC class I-restricted CD8-mediated protection against L. monocytogenes. The LLO+ B. anthracis recombinant strain represents a potential vector for delivering foreign Ags involved in CD8-mediated protective responses.


Are the uterine serous carcinomas underdiagnosed? Histomorphologic and immunohistochemical correlates and clinical follow up in high-grade endometrial carcinomas initially diagnosed as high-grade endometrioid carcinoma.
Histologic subclassification of high-grade endometrial carcinomas can sometimes be a diagnostic challenge when based on histomorphology alone. Here we utilized immunohistochemical markers to determine the immunophenotype in histologically ambiguous high-grade endometrial carcinomas that were initially diagnosed as pure or mixed high-grade endometrioid carcinoma, aiming to determine the utility of selected immunohistochemical panel in accurate classification of these distinct tumor types, while correlating these findings with the clinical outcome. A total of 43 high-grade endometrial carcinoma cases initially classified as pure high-grade endometrioid carcinoma (n=32), mixed high-grade endometrioid carcinoma/serous carcinoma (n=9) and mixed high-grade endometrioid carcinoma/clear cell carcinoma (n=2) were retrospectively stained with a panel of immunostains, including antibodies for p53, p16, estrogen receptor, and mammaglobin. Clinical follow-up data were obtained, and stage-to-stage disease outcomes were compared for different tumor types. Based on aberrant staining for p53 and p16, 17/43 (40%) of the high-grade endometrial carcinoma cases initially diagnosed as high-grade endometrioid carcinoma were re-classified as serous carcinoma. All 17 cases showed negative staining for mammaglobin, while estrogen receptor was positive in only 6 (35%) cases. The remaining 26 cases of high-grade endometrioid carcinoma showed wild-type staining for p53 in 25 (96%) cases, patchy staining for p16 in 20 (77%) cases, and were positive for mammaglobin and estrogen receptor in 8 (31%) and 19 (73%) cases, respectively, thus the initial diagnosis of high-grade endometrioid carcinoma was confirmed in these cases. In addition, the patients with re-classified serous carcinoma had advanced clinical stages at diagnosis and poorer overall survival on clinical follow-up compared to that of the remaining 26 high-grade endometrioid carcinoma cases. These results indicate that selected immunohistochemical panel, including p53, p16, and mammaglobin can be helpful in reaching accurate diagnosis in cases of histomorphologically ambiguous endometrial carcinomas, and can assist in providing guidance for appropriate therapeutic options for the patients.


Oculo-Visual Dysfunction in Parkinson's Disease.
This review describes the oculo-visual problems likely to be encountered in Parkinson's disease (PD) with special reference to three questions: (1) are there visual symptoms characteristic of the prodromal phase of PD, (2) is PD dementia associated with specific visual changes, and (3) can visual symptoms help in the differential diagnosis of the parkinsonian syndromes, viz. PD, progressive supranuclear palsy (PSP), dementia with Lewy bodies (DLB), multiple system atrophy (MSA), and corticobasal degeneration (CBD)? Oculo-visual dysfunction in PD can involve visual acuity, dynamic contrast sensitivity, colour discrimination, pupil reactivity, eye movement, motion perception, and visual processing speeds. In addition, disturbance of visuo-spatial orientation, facial recognition problems, and chronic visual hallucinations may be present. Prodromal features of PD may include autonomic system dysfunction potentially affecting pupil reactivity, abnormal colour vision, abnormal stereopsis associated with postural instability, defects in smooth pursuit eye movements, and deficits in visuo-motor adaptation, especially when accompanied by idiopathic rapid eye movement (REM) sleep behaviour disorder. PD dementia is associated with the exacerbation of many oculo-visual problems but those involving eye movements, visuo-spatial function, and visual hallucinations are most characteristic. Useful diagnostic features in differentiating the parkinsonian symptoms are the presence of visual hallucinations, visuo-spatial problems, and variation in saccadic eye movement dysfunction.


Compensation for dystrophin-deficiency: ADAM12 overexpression in skeletal muscle results in increased alpha 7 integrin, utrophin and associated glycoproteins.
Mouse models for genetic diseases are among the most powerful tools available for developing and testing new treatment strategies. ADAM12 is a disintegrin and metalloprotease, previously demonstrated to significantly alleviate the pathology of mdx mice, a model for Duchenne muscular dystrophy in humans. More specifically ADAM12 appeared to prevent muscle cell necrosis in the mdx mice as evidenced by morphological analysis and by the reduced levels of serum creatine kinase. In the present study we demonstrated that ADAM12 may compensate for the dystrophin deficiency in mdx mice by increasing the expression and redistribution of several components of the muscle cell-adhesion complexes. First, we analyzed transgenic mice that overexpress ADAM12 and found mild myopathic changes and accelerated regeneration following acute injury. We then analyzed changes in gene-expression profiles in mdx/ADAM12 transgenic mice compared with their littermate controls and found only a few genes with an expression change greater than 2-fold between mdx/ADAM12 and mdx. The small changes in gene expression were unexpected, considering the marked improvement of the mdx pathology when ADAM12 is overexpressed, and suggested that significant changes in mdx/ADAM12 muscle might occur post-transcriptionally. Indeed, by immunostaining and immunoblotting we found an approximately 2-fold increase in expression, and distinct extrasynaptic localization, of alpha 7B integrin and utrophin, the functional homolog of dystrophin. The expression of the dystrophin-associated glycoproteins was also increased. In conclusion, these results demonstrate a novel way to alleviate dystrophin deficiency in mice, and may stimulate the development of new approaches to compensate for dystrophin deficiency in animals and humans.


Apple miRNAs and tasiRNAs with novel regulatory networks.
MicroRNAs (miRNAs) and their regulatory functions have been extensively characterized in model species but whether apple has evolved similar or unique regulatory features remains unknown. We performed deep small RNA-seq and identified 23 conserved, 10 less-conserved and 42 apple-specific miRNAs or families with distinct expression patterns. The identified miRNAs target 118 genes representing a wide range of enzymatic and regulatory activities. Apple also conserves two TAS gene families with similar but unique trans-acting small interfering RNA (tasiRNA) biogenesis profiles and target specificities. Importantly, we found that miR159, miR828 and miR858 can collectively target up to 81 MYB genes potentially involved in diverse aspects of plant growth and development. These miRNA target sites are differentially conserved among MYBs, which is largely influenced by the location and conservation of the encoded amino acid residues in MYB factors. Finally, we found that 10 of the 19 miR828-targeted MYBs undergo small interfering RNA (siRNA) biogenesis at the 3' cleaved, highly divergent transcript regions, generating over 100 sequence-distinct siRNAs that potentially target over 70 diverse genes as confirmed by degradome analysis. Our work identified and characterized apple miRNAs, their expression patterns, targets and regulatory functions. We also discovered that three miRNAs and the ensuing siRNAs exploit both conserved and divergent sequence features of MYB genes to initiate distinct regulatory networks targeting a multitude of genes inside and outside the MYB family.


Screening for Language Disorders in Stroke: German Validation of the Language Screening Test (LAST).
Screening of aphasia in acute stroke is crucial for directing patients to early language therapy. The Language Screening Test (LAST), originally developed in French, is a validated language screening test that allows detection of a language deficit within a few minutes. The aim of the present study was to develop and validate two parallel German versions of the LAST. The LAST includes subtests for naming, repetition, automatic speech, and comprehension. For the translation into German, task constructs and psycholinguistic criteria for item selection were identical to the French LAST. A cohort of 101 stroke patients were tested, all of whom were native German speakers. Validation of the LAST was based on (1) analysis of equivalence of the German versions, which was established by administering both versions successively in a subset of patients, (2) internal validity by means of internal consistency analysis, and (3) external validity by comparison with the short version of the Token Test in another subset of patients. The two German versions were equivalent as demonstrated by a high intraclass correlation coefficient of 0.91. Furthermore, an acceptable internal structure of the LAST was found (Cronbach's α = 0.74). A highly significant correlation (r = 0.74, p < 0.0001) between the LAST and the short version of the Token Test indicated good external validity of the scale. The German version of the LAST, available in two parallel versions, is a new and valid language screening test in stroke.


Microcomputers and the future of epidemiology.
The Workshop on Microcomputers and the Future of Epidemiology was held March 8-9, 1993, at the Turner Conference Center, Atlanta, GA, with 130 public health professionals participating. The purpose of the workshop was to define microcomputer needs in epidemiology and to propose future initiatives. Thirteen groups representing public health disciplines defined their needs for better and more useful data, development of computer technology appropriate to epidemiology, user support and human infrastructure development, and global communication and planning. Initiatives proposed were demonstration of health surveillance systems, new software and hardware, computer-based training, projects to establish or improve data bases and community access to data bases, improved international communication, conferences on microcomputer use in particular disciplines, a suggestion to encourage competition in the production of public-domain software, and longrange global planning for epidemiologic computing and data management. Other interested groups are urged to study, modify, and implement those ideas.


Tailored Education to Increase Self-Efficacy for Caregivers of Children With Asthma: A Randomized Controlled Trial.
We examined the efficacy of a tailored education program using touch-screen computers for caregivers of young children with asthma, aged 0 to 6 years. Fifty-three caregivers (mean age, 36.54 years; 51 mothers and two fathers) were recruited during typical visits to a national children's hospital. Caregivers were assigned randomly to 28 in the tailored education or 25 in the booklet education groups. The tailored education group received messages generated by a computer program, and the researchers provided them with counseling. The booklet education group only received an educational booklet. The primary outcome variable was parental self-efficacy and its subscales (ie, medication behavior, environmental behavior, barriers to self-management, and consultation behavior). The tailored education group showed an increase in barriers to management and asthma knowledge compared with the booklet education group. Over time, caregivers' quality of life improved significantly in both groups. Tailored education programs for caregivers of asthmatic children require further educational content modifications to match the learners' characteristics more closely. This study supports the notion that the development of patient education with tailored computer programs improves parental quality of life of asthma.


WHO expert committee on specifications for pharmaceutical preparations.
The Expert Committee on Specifications for Pharmaceutical Preparations works towards clear, independent and practical standards and guidelines for the quality assurance of medicines. Standards are developed by the Committee through worldwide consultation and an international consensus-building process. The following new guidelines were adopted and recommended for use: Release procedure for International Chemical Reference Substances; WHO guidelines on quality risk management; WHO guidelines on variations to a prequalified product; and the Collaborative procedure between the World Health Organization Prequalification of Medicines Programme and national medicines regulatory authorities in the assessment and accelerated national registration of WHO-prequalified pharmaceutical products.


Crystal structure of Paracoccus denitrificans electron transfer flavoprotein: structural and electrostatic analysis of a conserved flavin binding domain.
The crystal structure of electron transfer flavoprotein (ETF) from Paracoccus denitrificans was determined and refined to an R-factor of 19.3% at 2.6 A resolution. The overall fold is identical to that of the human enzyme, with the exception of a single loop region. Like the human structure, the structure of the P. denitrificans ETF is comprised of three distinct domains, two contributed by the alpha-subunit and the third from the beta-subunit. Close analysis of the structure reveals that the loop containing betaI63 is in part responsible for conferring the high specificity of AMP binding by the ETF protein. Using the sequence and structures of the human and P. denitrificans enzymes as models, a detailed sequence alignment has been constructed for several members of the ETF family, including sequences derived for the putative FixA and FixB proteins. From this alignment, it is evident that in all members of the ETF family the residues located in the immediate vicinity of the FAD cofactor are identical, with the exception of the substitution of serine and leucine residues in the W3A1 ETF protein for the human residues alphaT266 and betaY16, respectively. Mapping of ionic differences between the human and P. denitrificans ETF onto the structure identifies a surface that is electrostatically very similar between the two proteins, thus supporting a previous docking model between human ETF and pig medium-chain acyl-CoA dehydrogenase (MCAD). Analysis of the ionic strength dependence of the electron transfer reaction between either human or P. denitrificans ETF and MCAD demonstrates that the human ETF functions optimally at low ( approximately 10 mequiv) ionic strength, while P. denitrificans ETF is a better electron acceptor at higher (>75 mequiv) ionic strength. This suggests that the electrostatic surface potential of the two proteins is very different and is consistent with the difference in isoelectric points between the proteins. Analysis of the electrostatic potentials of the human and P. denitrificans ETFs reveals that the P. denitrificans ETF is more negatively charged. This excess negative charge may contribute to the difference in redox potentials between the two ETF flavoproteins and suggests an explanation for the opposing ionic strength dependencies for the reaction of MCAD with the two ETFs. Furthermore, by analysis of a model of the previously described human-P. denitrificans chimeric ETF protein, it is possible to identify one region of ETF that participates in docking with ETF-ubiquinone oxidoreductase, the physiological electron acceptor for ETF.


Tumour-infiltrating CD8+ lymphocytes and colorectal cancer recurrence by tumour and nodal stage.
Intratumoural T-cell infiltrate intensity cortes wrelaith clinical outcome in stage II/III colorectal cancer (CRC). We aimed to determine whether this association varies across this heterogeneous group. We performed a pooled analysis of 1804 CRCs from the QUASAR2 and VICTOR trials. Intratumoural CD8+ and CD3+ densities were quantified by immunohistochemistry in tissue microarray (TMA) cores, and their association with clinical outcome analysed by Cox regression. We validated our results using publicly available gene expression data in a pooled analysis of 1375 CRCs from seven independent series. In QUASAR2, intratumoural CD8+ was a stronger predictor of CRC recurrence than CD3+ and showed similar discriminative ability to both markers in combination. Pooled multivariable analysis of both trials showed increasing CD8+ density was associated with reduced recurrence risk independent of confounders including DNA mismatch repair deficiency, POLE mutation and chromosomal instability (multivariable hazard ratio [HR] for each two-fold increase = 0.92, 95%CI = 0.87-0.97, P = 3.6 × 10-3). This association was not uniform across risk strata defined by tumour and nodal stage: absent in low-risk (pT3,N0) cases (HR = 1.03, 95%CI = 0.87-1.21, P = 0.75), modest in intermediate-risk (pT4,N0 or pT1-3,N1-2) cases (HR = 0.92, 95%CI = 0.86-1.0, P = 0.046) and strong in high-risk (pT4,N1-2) cases (HR = 0.87, 95%CI = 0.79-0.97, P = 9.4 × 10-3); PINTERACTION = 0.090. Analysis of tumour CD8A expression in the independent validation cohort revealed similar variation in prognostic value across risk strata (PINTERACTION = 0.048). The prognostic value of intratumoural CD8+ cell infiltration in stage II/III CRC varies across tumour and nodal risk strata.


Frequency of incomplete medication histories obtained at triage.
The frequency of incomplete medication histories obtained at triage in an emergency department (ED) is described. The survey of medication histories collected during ED triage was conducted during a 20-week period. Data collection occurred on weekdays during the dayshift for 15 hours per week for a total of 300 hours. Patients who bypassed triage or were unconscious, unable to communicate, uncooperative, or violent were excluded. Ten student pharmacists were trained on study procedures and collected data using a data collection tool which included patient's chief complaints, medications and dosages, and whether medications were identified at triage. Patients' medication-related ED visits were classified as being caused by adverse effects, medication errors, poor adherence, intentional overdose, or therapeutic failure. During the 300 hours of data collection, 2063 patients were admitted to the ED. Of these, 1465 (71%) were interviewed and evaluated for complete medication histories. Among 1172 (80%) patients identified as taking medications, the history obtained at triage failed to identify at least one medication in 707 (48%) patients. In cases where medications were not identified, a mixture of prescription (73%) and nonprescription (27%) medications were missed with a median of 2 drugs (range, 1-20 drugs). Drugs missed at triage were related to the patient's chief complaint in 27% of the cases. Medication histories collected at triage in the ED of an urban medical center were often incomplete, especially among patients taking multiple medications. Efforts should be taken to improve methods for obtaining more complete medication histories during triage and collecting supplemental medication histories to ensure appropriate emergency care.


Continuous focusing of microparticles using inertial lift force and vorticity via multi-orifice microfluidic channels.
We developed a new microfluidic method for focusing microparticles through the combined use of inertial lift forces and turbulent secondary flows generated in a topographically patterned microchannel. The mechanism of particle focusing is based on the hydrodynamic inertial forces exerted on particles migrating along a non-circular microchannel, i.e.tubular pinch effect and wall effect, which induce particle movement away from walls and along a specific lateral position in the microchannel. With the extraordinary geometry of multi-orifice microchannel, an ordered and focused particle distribution was achieved at central or side regions according to a particle Reynolds number (Re(p)) range. The focusing of particles was controlled by the particle Reynolds number, microchannel length, and volume fraction of particles in suspension. This method will be beneficial in particle focusing processes in a microfluidic device since it offers continuous, high-throughput performance and simple operation.


Disentangling Sources of Gene Tree Discordance in Phylogenomic Data Sets: Testing Ancient Hybridizations in Amaranthaceae s.l.
Gene tree discordance in large genomic data sets can be caused by evolutionary processes such as incomplete lineage sorting and hybridization, as well as model violation, and errors in data processing, orthology inference, and gene tree estimation. Species tree methods that identify and accommodate all sources of conflict are not available, but a combination of multiple approaches can help tease apart alternative sources of conflict. Here, using a phylotranscriptomic analysis in combination with reference genomes, we test a hypothesis of ancient hybridization events within the plant family Amaranthaceae s.l. that was previously supported by morphological, ecological, and Sanger-based molecular data. The data set included seven genomes and 88 transcriptomes, 17 generated for this study. We examined gene-tree discordance using coalescent-based species trees and network inference, gene tree discordance analyses, site pattern tests of introgression, topology tests, synteny analyses, and simulations. We found that a combination of processes might have generated the high levels of gene tree discordance in the backbone of Amaranthaceae s.l. Furthermore, we found evidence that three consecutive short internal branches produce anomalous trees contributing to the discordance. Overall, our results suggest that Amaranthaceae s.l. might be a product of an ancient and rapid lineage diversification, and remains, and probably will remain, unresolved. This work highlights the potential problems of identifiability associated with the sources of gene tree discordance including, in particular, phylogenetic network methods. Our results also demonstrate the importance of thoroughly testing for multiple sources of conflict in phylogenomic analyses, especially in the context of ancient, rapid radiations. We provide several recommendations for exploring conflicting signals in such situations. [Amaranthaceae; gene tree discordance; hybridization; incomplete lineage sorting; phylogenomics; species network; species tree; transcriptomics.].


Cerebrotendinous xanthomatosis in three siblings from a Chinese family.
Cerebrotendinous xanthomatosis (CTX) is exceptionally rare in the Chinese population. We reported a 31-year-old Chinese male in Hong Kong, who has the characteristic features of cerebrotendinous xanthomatosis including the multiple xanthomas of tendons, mental retardation, bilateral cataracts, cerebellar ataxia and spasticity of the left arm, high concentrations of plasma phytosterols and abnormal MR of brain. On screening the family, two other siblings of 27 and 29 respectively, have tendon xanthomas and high plasma phytosterols. An extensive search of the international medical literature, including the Medline, has revealed only one other case report of cerebrotendinous xanthomatosis in Taiwan. CTX is a potentially treatable disease. It is hoped by alertness, early diagnosis and treatment can be made, and hence prevent further progression of the disease.


[Pharmacopoeia in ancient Egypt].
Ancient Egyptian pharmacological science is high part of the Egyptian medicine, as evidenced by the so-called pharmacological papyrus Ebers, which contains prescriptions with different kinds of components of vegetable, mineral and animal origin. These data suggest that Egyptian pharmacopeia had an exceptional complexity in the whole of ancient medicine. Egyptian medical knowledge influenced Mediterranean civilizations, as evidenced not only by the Greek translations of names of components but also by the Egyptian and Egyptian-like prescriptions found in the Corpus Hippocraticum.


Intellectual, behavioral, and emotional functioning in children with syndromic craniosynostosis.
To examine intellectual, behavioral, and emotional functioning of children who have syndromic craniosynostosis and to explore differences between diagnostic subgroups. A national sample of children who have syndromic craniosynostosis participated in this study. Intellectual, behavioral, and emotional outcomes were assessed by using standardized measures: Wechsler Intelligence Scale for Children, Third Edition, Child Behavior Checklist (CBCL)/6-18, Disruptive Behavior Disorder rating scale (DBD), and the National Institute of Mental Health Diagnostic Interview Schedule for Children. We included 82 children (39 boys) aged 6 to 13 years who have syndromic craniosynostosis. Mean Full-Scale IQ (FSIQ) was in the normal range (M = 96.6; SD = 21.6). However, children who have syndromic craniosynostosis had a 1.9 times higher risk for developing intellectual disability (FSIQ < 85) compared with the normative population (P < .001) and had more behavioral and emotional problems compared with the normative population, including higher scores on the CBCL/6-18, DBD Total Problems (P < .001), Internalizing (P < .01), social problems (P < .001), attention problems (P < .001), and the DBD Inattention (P < .001). Children who have Apert syndrome had lower FSIQs (M = 76.7; SD = 13.3) and children who have Muenke syndrome had more social problems (P < .01), attention problems (P < .05), and inattention problems (P < .01) than normative population and with other diagnostic subgroups. Although children who have syndromic craniosynostosis have FSIQs similar to the normative population, they are at increased risk for developing intellectual disability, internalizing, social, and attention problems. Higher levels of behavioral and emotional problems were related to lower levels of intellectual functioning.


Multi-serotype pneumococcal nasopharyngeal carriage prevalence in vaccine naïve Nepalese children, assessed using molecular serotyping.
Invasive pneumococcal disease is one of the major causes of death in young children in resource poor countries. Nasopharyngeal carriage studies provide insight into the local prevalence of circulating pneumococcal serotypes. There are very few data on the concurrent carriage of multiple pneumococcal serotypes. This study aimed to identify the prevalence and serotype distribution of pneumococci carried in the nasopharynx of young healthy Nepalese children prior to the introduction of a pneumococcal conjugate vaccine using a microarray-based molecular serotyping method capable of detecting multi-serotype carriage. We conducted a cross-sectional study of healthy children aged 6 weeks to 24 months from the Kathmandu Valley, Nepal between May and October 2012. Nasopharyngeal swabs were frozen and subsequently plated on selective culture media. DNA extracts of plate sweeps of pneumococcal colonies from these cultures were analysed using a molecular serotyping microarray capable of detecting relative abundance of multiple pneumococcal serotypes. 600 children were enrolled into the study: 199 aged 6 weeks to <6 months, 202 aged 6 months to < 12 months, and 199 aged 12 month to 24 months. Typeable pneumococci were identified in 297/600 (49.5%) of samples with more than one serotype being found in 67/297 (20.2%) of these samples. The serotypes covered by the thirteen-valent pneumococcal conjugate vaccine were identified in 44.4% of samples containing typeable pneumococci. Application of a molecular serotyping approach to identification of multiple pneumococcal carriage demonstrates a substantial prevalence of co-colonisation. Continued surveillance utilising this approach following the introduction of routine use of pneumococcal conjugate vaccinates in infants will provide a more accurate understanding of vaccine efficacy against carriage and a better understanding of the dynamics of subsequent serotype and genotype replacement.


Characterization of a complex rearrangement involving chromosomes 1, 4 and 8 by FISH and array-CGH.
Complex chromosomal rearrangements (CCRs) are structural aberrations involving more than two chromosomes with at least three breakpoints. CCRs can be divided into familial and de novo. Balanced CCR are extremely rare in humans and are at high risk of producing unbalanced gametes. Individuals with balanced CCR are usually phenotipically normal but report fertility problems, recurrent miscarriages or congenital anomalies in newborn offsprings as consequence of either meiotic failure or imbalanced chromosomes segregation.We describe the case of an unbalanced CCR involving chromosomes 1, 4 and 8 found in a girl with developmental delay, hexadactilia and microcephaly. The rearrangement, apparently balanced at a standard karyotype analysis and of maternal origin, was demonstrated to be unbalanced by array-CGH and FISH. In conclusion our study underlines the importance of the combined use of a quantitative technique, as array-CGH, to detect criptic segmental aneuploidies, and a qualitative tool, as FISH analysis, to physically map the localization of the chromosome segments involved, in order to realize the exact nature that underlies a chromosomal rearrangement.


Lung neuroendocrine tumors: correlation of ubiquitinylation and sumoylation with nucleo-cytosolic partitioning of PTEN.
The tumor suppressor phosphatase and tensin homolog (PTEN) is a pleiotropic enzyme, inhibiting phosphatidyl-inositol-3 kinase (PI3K) signaling in the cytosol and stabilizing the genome in the nucleus. Nucleo-cytosolic partitioning is dependent on the post-translational modifications ubiquitinylation and sumoylation. This cellular compartmentalization of PTEN was investigated in lung neuroendocrine tumors (lung NET). Tumor tissues from 192 lung NET patients (surgical specimens = 183, autopsies = 9) were investigated on tissue microarrays. PTEN was H-scored by two investigators in nucleus and cytosol using the monoclonal antibody 6H2.1. Results were correlated with immunoreactivity for USP7 (herpes virus-associated ubiquitin-specific protease 7) and SUMO2/3 (small ubiquitin-related modifier protein 2/3) as well as PTEN and p53 FISH gene status. Clinico-pathologic data including overall survival, proliferation rate and diagnostic markers (synaptophysin, chromogranin A, Mib-1, TTF-1) were recorded. The multicentre cohort included 58 typical carcinoids (TC), 42 atypical carcinoids (AC), 32 large cell neuroendocrine carcinomas (LCNEC) and 60 small cell lung carcinomas (SCLC). Carcinoids were smaller in size and had higher synaptophysin and chromogranin A, but lower TTF-1 expressions. Patients with carcinoids were predominantly female and 10 years younger than patients with LCNEC/SCLC. In comparison to the carcinoids, LCNEC/SCLC tumors presented a stronger loss of nuclear and cytosolic PTEN associated with a loss of PTEN and p53. Concomitantly, a loss of nuclear USP7 but increase of nuclear and cytosolic SUMO2/3 was found. Loss of nuclear and cytosolic PTEN, loss of nuclear USP7 and increase of cytosolic SUMO2/3 thus correlated with poor survival. Among carcinoids, loss of cytosolic PTEN was predominantly found in TTF1-negative larger tumors of male patients. Among SCLC, loss of both cytosolic and nuclear PTEN but not proliferation rate or tumor size delineated a subgroup with poorer survival (all p-values <0.05). Cellular ubiquitinylation and sumoylation likely influence the functional PTEN loss in high grade lung NET. Both nuclear and cytosolic PTEN immunoreactivity should be considered for correlation with clinico-pathologic parameters.


Indirect cholinergic activation slows down pancreatic cancer growth and tumor-associated inflammation.
Nerve-cancer interactions are increasingly recognized to be of paramount importance for the emergence and progression of pancreatic cancer (PCa). Here, we investigated the role of indirect cholinergic activation on PCa progression through inhibition of acetylcholinesterase (AChE) via clinically available AChE-inhibitors, i.e. physostigmine and pyridostigmine. We applied immunohistochemistry, immunoblotting, MTT-viability, invasion, flow-cytometric-cell-cycle-assays, phospho-kinase arrays, multiplex ELISA and xenografted mice to assess the impact of AChE inhibition on PCa cell growth and invasiveness, and tumor-associated inflammation. Survival analyses were performed in a novel genetically-induced, surgically-resectable mouse model of PCa under adjuvant treatment with gemcitabine+/-physostigmine/pyridostigmine (n = 30 mice). Human PCa specimens (n = 39) were analyzed for the impact of cancer AChE expression on tumor stage and survival. We discovered a strong expression of AChE in cancer cells of human PCa specimens. Inhibition of this cancer-cell-intrinsic AChE via pyridostigmine and physostigmine, or administration of acetylcholine (ACh), diminished PCa cell viability and invasion in vitro and in vivo via suppression of pERK signaling, and reduced tumor-associated macrophage (TAM) infiltration and serum pro-inflammatory cytokine levels. In the novel genetically-induced, surgically-resectable PCa mouse model, adjuvant co-therapy with AChE blockers had no impact on survival. Accordingly, survival of resected PCa patients did not differ based on tumor AChE expression levels. Patients with higher-stage PCa also exhibited loss of the ACh-synthesizing enzyme, choline-acetyltransferase (ChAT), in their nerves. For future clinical trials of PCa, direct cholinergic stimulation of the muscarinic signaling, rather than indirect activation via AChE blockade, may be a more effective strategy.


Notch4 normalization reduces blood vessel size in arteriovenous malformations.
Abnormally enlarged blood vessels underlie many life-threatening disorders including arteriovenous (AV) malformations (AVMs). The core defect in AVMs is high-flow AV shunts, which connect arteries directly to veins, "stealing" blood from capillaries. Here, we studied mouse brain AV shunts caused by up-regulation of Notch signaling in endothelial cells (ECs) through transgenic expression of constitutively active Notch4 (Notch4*). Using four-dimensional two-photon imaging through a cranial window, we found that normalizing Notch signaling by repressing Notch4* expression converted large-caliber, high-flow AV shunts to capillary-like vessels. The structural regression of the high-flow AV shunts returned blood to capillaries, thus reversing tissue hypoxia. This regression was initiated by vessel narrowing without the loss of ECs and required restoration of EphB4 receptor expression by venous ECs. Normalization of Notch signaling resulting in regression of high-flow AV shunts, and a return to normal blood flow suggests that targeting the Notch pathway may be useful therapeutically for treating diseases such as AVMs.


FLEXBAR-Flexible Barcode and Adapter Processing for Next-Generation Sequencing Platforms.
Quantitative and systems biology approaches benefit from the unprecedented depth of next-generation sequencing. A typical experiment yields millions of short reads, which oftentimes carry particular sequence tags. These tags may be: (a) specific to the sequencing platform and library construction method (e.g., adapter sequences); (b) have been introduced by experimental design (e.g., sample barcodes); or (c) constitute some biological signal (e.g., splice leader sequences in nematodes). Our software FLEXBAR enables accurate recognition, sorting and trimming of sequence tags with maximal flexibility, based on exact overlap sequence alignment. The software supports data formats from all current sequencing platforms, including color-space reads. FLEXBAR maintains read pairings and processes separate barcode reads on demand. Our software facilitates the fine-grained adjustment of sequence tag detection parameters and search regions. FLEXBAR is a multi-threaded software and combines speed with precision. Even complex read processing scenarios might be executed with a single command line call. We demonstrate the utility of the software in terms of read mapping applications, library demultiplexing and splice leader detection. FLEXBAR and additional information is available for academic use from the website: http://sourceforge.net/projects/flexbar/.


Serum neurofilament light chain in pediatric MS and other acquired demyelinating syndromes.
To explore the correlation between serum and CSF neurofilament light chain (NfL) and the association of NfL levels and future disease activity in pediatric patients with a first attack of acquired demyelinating syndromes (ADS). In total, 102 children <18 years with a first attack of CNS demyelination and 23 age-matched controls were included. Clinically definite multiple sclerosis (CDMS) was set as an endpoint for analysis. CSF NfL was tested by the commercially available ELISA (UmanDiagnostics); serum NfL (sNfL) was tested with a Simoa assay. Hazard ratios (HR) were calculated with Cox regression analysis. Of the 102 patients, 47 (46%) were tested for CSF NfL. CSF and serum NfL correlated significantly in the total group (ρ 0.532, p < 0.001) and even more significantly in the subgroup of patients with future CDMS diagnosis (ρ 0.773, p < 0.001). sNfL was higher in patients than in controls (geometric mean 6.1 pg/mL, p < 0.001), and was highest in ADS presenting with encephalopathy (acute disseminated encephalomyelitis, n = 28, 100.4 pg/mL), followed by patients without encephalopathy (ADS-) with future CDMS diagnosis (n = 40, 32.5 pg/mL), and ADS- who remained monophasic (n = 34, 17.6 pg/mL). sNfL levels higher than a median of 26.7 pg/mL at baseline are associated with a shorter time to CDMS diagnosis in ADS- (p = 0.045). HR for CDMS diagnosis was 1.09 for each 10 pg/mL increase of sNfL, after correction for age, oligoclonal bands, and MRI measures (p = 0.012). The significant correlation between CSF and serum NfL strengthens its reliability as a peripheral marker of neuroaxonal damage. Higher sNfL levels at baseline were associated with higher probability of future CDMS diagnosis in ADS-.


Identifying the regional-scale groundwater-surface water interaction on the Sanjiang Plain, Northeast China.
Assessment on the interaction between groundwater and surface water (GW-SW) can generate information that is critical to regional water resource management, especially for regions that are highly dependent on groundwater resources for irrigation. This study investigated such interaction on China's Sanjiang Plain (10.9 × 10(4) km(2)) and produced results to assist sustainable regional water management for intensive agricultural activities. Methods of hierarchical cluster analysis (HCA), principal component analysis (PCA), and statistical analysis were used in this study. One hundred two water samplings (60 from shallow groundwater, 7 from deep groundwater, and 35 from surface water) were collected and grouped into three clusters and seven sub-clusters during the analyses. The PCA analysis identified four principal components of the interaction, which explained 85.9% variance of total database, attributed to the dissolution and evolution of gypsum, feldspar, and other natural minerals in the region that was affected by anthropic and geological (sedimentary rock mineral) activities. The analyses showed that surface water in the upper region of the Sanjiang Plain gained water from local shallow groundwater, indicating that the surface water in the upper region was relatively more resilient to withdrawal for usage, whereas in the middle region, there was only a weak interaction between shallow groundwater and surface water. In the lower region of the Sanjiang Plain, surface water lost water to shallow groundwater, indicating that the groundwater was vulnerable to pollution by pesticides and fertilizers from terrestrial sources.


Epidemiology of chronic pain and its treatment in The Netherlands.
Chronic pain is common; however, good epidemiological data are scarce. Such information can help all the involved stakeholders to make responsible decisions about health budgets and prioritisation. This study aims to provide best-evidence epidemiological information about chronic pain in the Netherlands. We performed a systematic search which yielded 16,619 references, 119 Dutch studies were relevant. We selected at least three studies per question that provided the most recent, representative and valid data. The prevalence of moderate to severe general chronic pain among Dutch adults was estimated at 18%. This prevalence was 27% and 55% for any cancer pain. Up to 74% of patients with general or non-cancer chronic pain get treated; this percentage is little higher for patients with cancer pain. A substantial proportion of the patients receive drug treatment for their pain, mainly NSAIDs, but also non-pharmacological interventions for pain are being used. Up to 43% of the chronic non-cancer pain patients report not receiving treatment and up to 79% of the patients believe their pain is inadequately treated. All studies reported a detrimental effect of chronic pain on quality of life, activities of daily living and mental health. Chronic pain is also associated with direct and indirect medical costs, and patients may have decreased income and additional out-of pocket expenses. Chronic pain occur s frequently, has a negative impact on the patient and society and treatment may not always be adequate. Chronic pain should be seen as an important public health problem deserving more attention of Dutch healthcare workers and policy makers.


Transport of bile acids, sulfated steroids, estradiol 17-beta-D-glucuronide, and leukotriene C4 by human multidrug resistance protein 8 (ABCC11).
We previously determined that expression of human multidrug resistance protein (MRP) 8, a recently described member of the MRP family of ATP-binding cassette transporters, enhances cellular extrusion of cyclic nucleotides and confers resistance to nucleotide analogs (J Biol Chem 278:29509-29514, 2003). However, the in vitro transport characteristics of the pump have not been determined. In this study, the substrate selectivity and biochemical activity of MRP8 is investigated using membrane vesicles prepared from LLC-PK1 cells transfected with MRP8 expression vector. Expression of MRP8 is shown to stimulate the ATP-dependent uptake of a range of physiological and synthetic lipophilic anions, including the glutathione S-conjugates leukotriene C4 and dinitrophenyl S-glutathione, steroid sulfates such as dehydroepiandrosterone 3-sulfate (DHEAS) and estrone 3-sulfate, glucuronides such as estradiol 17-beta-D-glucuronide (E(2)17betaG), the monoanionic bile acids glycocholate and taurocholate, and methotrexate. In addition, MRP8 is competent in the in vitro transport of cAMP and cGMP, in accord with the results of our previously reported cellular studies. DHEAS, E(2)17betaG, and methotrexate were transported with K(m) and V(max) values of 13.0 +/- 0.8 microM and 34.9 +/- 9.5 pmol/mg/min, 62.9 +/- 12 microM and 62.0 +/- 5.2 pmol/mg/min, and 957 +/- 28 microM and 317 +/- 17 pmol/mg/min, respectively. Based upon the stimulatory action of DHEAS on uptake of E(2)17betaG, the attenuation of this effect at high DHEAS concentrations and the lack of reciprocal promotion of DHEAS uptake by E(2)17betaG, a model involving nonreciprocal constructive interactions between some transport substrates is invoked. These results suggest that MRP8 participates in physiological processes involving bile acids, conjugated steroids, and cyclic nucleotides and indicate that the pump has complex interactions with its substrates.


Deregulation of HEF1 impairs M-phase progression by disrupting the RhoA activation cycle.
The focal adhesion-associated signaling protein HEF1 undergoes a striking relocalization to the spindle at mitosis, but a function for HEF1 in mitotic signaling has not been demonstrated. We here report that overexpression of HEF1 leads to failure of cells to progress through cytokinesis, whereas depletion of HEF1 by small interfering RNA (siRNA) leads to defects earlier in M phase before cleavage furrow formation. These defects can be explained mechanistically by our determination that HEF1 regulates the activation cycle of RhoA. Inactivation of RhoA has long been known to be required for cytokinesis, whereas it has recently been determined that activation of RhoA at the entry to M phase is required for cellular rounding. We find that increased HEF1 sustains RhoA activation, whereas depleted HEF1 by siRNA reduces RhoA activation. Furthermore, we demonstrate that chemical inhibition of RhoA is sufficient to reverse HEF1-dependent cellular arrest at cytokinesis. Finally, we demonstrate that HEF1 associates with the RhoA-GTP exchange factor ECT2, an orthologue of the Drosophila cytokinetic regulator Pebble, providing a direct means for HEF1 control of RhoA. We conclude that HEF1 is a novel component of the cell division control machinery and that HEF1 activity impacts division as well as cell attachment signaling events.


Diuretic treatment alters clonidine suppression of plasma norepinephrine.
The effect of short-term diuretic treatment on the action of clonidine was evaluated in eight subjects with mild, uncomplicated hypertension. A single oral dose of clonidine (0.3 mg) was given before and after 1 week of therapy with hydrochlorothiazide, 50 mg, and amiloride, 5 mg, taken daily. Changes in mean arterial pressure, heart rate, plasma norepinephrine and epinephrine levels, and plasma renin activity were assessed. Diuretic treatment caused a significant weight loss, increased plasma renin activity, and reduced serum potassium concentration but did not significantly alter the absolute reduction in mean arterial pressure caused by clonidine. Absolute clonidine-induced reduction in plasma renin activity after diuretic treatment was three times greater than before treatment, although percent changes were similar. Before diuretic therapy, clonidine significantly reduced the level of norepinephrine (absolute and percent change). After diuretic treatment, clonidine failed to suppress norepinephrine, and the difference from prediuretic changes was significant. The level of epinephrine was not altered significantly either by diuretic treatment or clonidine. These results indicate that diuretic therapy alters the clonidine-activated mechanism for reduction of arterial pressure through a shift from overall suppression of sympathetic tone to pathways that are more restricted to renal tone. This shift may be due to changes in fluid or electrolyte balance that alter the action of alpha 2-adrenergic receptor-mediated pathways. Use of the clonidine suppression test for the diagnosis of pheochromocytoma may give false-positive results in diuretic-treated patients.


"It's Just Presence," the Contributions of Aboriginal and Torres Strait Islander Health Professionals in Cancer Care in Queensland.
Objectives: The aim of this research was to explore health professionals' perspectives on the provision of follow-up cancer care for Aboriginal and Torres Strait Islander patients in Queensland. Methods: Semi-structured interviews were conducted with Indigenous and non-Indigenous health professionals who had experience providing care for Indigenous cancer patients in the primary health care and hospital setting. Results: Participants were recruited from six Aboriginal Community Controlled Health Services (n = 17) and from a tertiary hospital (n = 9) across urban, regional, and remote geographical settings. Culturally safe care, psychological support, determining patient needs, practical assistance, and advocating for Indigenous health were identified as enablers to support the needs of Indigenous patients when accessing cancer care, and Indigenous health professionals were identified as the key enabler. Conclusion: Indigenous health professionals significantly contribute to the provision of culturally competent follow-up cancer care by increasing the accessibility of follow-up cancer care services and by supporting the needs of Indigenous cancer patients. All health professionals need to work together and be sufficiently skilled in the delivery of culturally competent care to improve the Indigenous cancer journey and outcomes for Indigenous people. Effective organizational policies and practices are crucial to enable all health professionals to provide culturally competent and responsive cancer care to Indigenous Australians.


The selection of skin care products for use in hyperbaric chamber may depend on flammability acceptability indices score.
Current protocols call for stopping adjunctive skin care treatments during hyperbaric oxygen therapy (HBOT) because the hyperbaric environment is considered unsafe for skin care products. The elevated oxygen fraction and the increased pressure in the hyperbaric chamber dramatically increase the flammability potential of materials, leading to the need for rigorous standards to prevent flame ignition. A scientific method of evaluating the flammability risks associated with skin care products would be helpful. Several skin care products were tested, using established industrial techniques for determining flammability potential with some modification. The information obtained from these tests can help clinicians make more rational decisions about which topical products can be used safely on patients undergoing HBOT. Wendell Hull & Associates conducted independent studies, comparing the oxygen compatibility for leading skin care products. Oxygen compatibility was determined using autogenous ignition temperature (AIT), oxygen index (OI), and heat of combustion (HoC) testing. AIT, a relative indication of a material's propensity for ignition, is the minimum temperature needed to cause a sample to self-ignite at a given pressure and oxygen concentration. OI, a relative indication of a material's flammability, is the minimum oxygen percentage that, when mixed with nitrogen, will sustain burning. HoC is the absolute value of a material's energy release when burning, if ignition occurs. Products with a high AIT, a high OI, and a low HoC are more compatible in an oxygen-enriched atmosphere (OEA). An acceptability index (AI) based on these 3 factors was calculated for the products, so the testers could rank overall material compatibility in OEAs (Lapin A. Oxygen Compatibility of Materials. International Institute of Refrigeration Commission Meeting; Brighton, England; 1973). Test results for the skin products varied widely. The AIT, OI, HoC, and AI were determined for each product under described circumstances. The AIT results indicate that all products in 99.5% oxygen concentration under pressure will ignite and that a pattern based on the absence or presence of petroleum-based ingredients does not seem to exist. Products containing petrolatum, mineral oil, paraffin, and paraffin wax had a HoC that equaled or exceeded the HoC of gasoline, whereas products without petroleum-based ingredients had a significantly lower HoC. The OI of skin products not containing petrolateum-based ingredients was significantly higher than the OI of products containing it. The AI values the OI as the most important value: the higher the AI, the more acceptable the product is for use with oxygen. The silicone-containing, petroleum-free products received an AI up to 25 times higher than the petrolatum-based products. These findings suggest a wide variation in the safety profiles of skin products. Skin products being considered for use in an OEA should be screened for flammability risks. This screening will allow informed decisions about the fire safety of the products. Further research is indicated.


[Changes in the diet pattern of the Brazilian urban population (1962-1988)].
Contemporary changes in food consumption patterns in urban areas of Brazil are described. The main data sources are two national food expenditure surveys undertaken in the early 60's and late 80's (1961-62 and 1987-88) and one national food intake survey undertaken in the mid-70's (1974-75). The analysis is restricted to the seven metropolitan areas represented in the three surveys. Food patterns are described on the basis of the relative consumption of different food groups, proportion of energy from carbohydrates, protein and lipids, proportion of animal and vegetable protein and fats, proportion of complex and simple carbohydrates, ratio between polyunsaturated and saturated fatty acids and relative consumption of cholesterol. Main food changes were similar in the Northeastern and Southeastern cities and involved: 1) reduction in the relative consumption of cereals, beans, roots and tubercles; 2) replacement of lard, bacon and butter by vegetable oils and margarine: and 3) increase in the relative consumption of eggs, milk and dairy products. As a net result of these changes there was, simultaneously, a decrease in the carbohydrate content of the Brazilian diet and an increase in its fat content. Total protein content of the diet remained at around 12% in the three surveys but animal protein increased. Changes in the fat content of the diet involved an increased proportion of vegetable fat, an increased ratio of poly-unsaturated/saturated fatty acids and a reduction in cholesterol intake. Health implications of dietary changes are discussed based on the World Health Organization's dietary guidelines.


Practical aspects of radiation safety for using fluorine-18.
The use of positron-emitting nuclides is becoming routine in nuclear medicine departments today. Introducing these nuclides into the nuclear medicine department can be a smooth transition by instituting educational lectures, radiation safety protocols and patient education. The radiation safety concerns of the technical staff, physicians and ancillary personnel are important and must be addressed. Nuclear medicine departments can be optimistic about implementing PET imaging while staying well within ALARA guidelines. After reading this article, the technologist should be able to: (a) describe at least three ways to reduce the radiation dose to the technologist during the performance of PET imaging procedures with 18F; (b) discuss the relationships between gamma-ray energy, the amount of activity administered to a patient, exposure time and occupational dose; and (c) describe one strategy to minimize the radiation dose to the bladder in patients who have received 18F.


Comprehensive studies of the Li+ effect on NaYF4:Yb/Er nanocrystals: morphology, structure, and upconversion luminescence.
Impurity doping plays a critical role in altering the properties of target nanomaterials in terms of designed morphologies, crystal structures, and functionalities. In this work, we have performed a comprehensive investigation of the effect of Li+ doping on the morphology, crystal structure, and upconversion luminescence of NaYF4:Yb/Er nanocrystals. Different Li+ sources, e.g., LiOA and LiOH, were used and the Li+ doping concentration varied from 0 to 100 mol%. The final product changes from hexagonal NaYF4:Yb/Er to the mixture of cubic NaYF4:Yb/Er and tetragonal LiYF4:Yb/Er, and finally to pure tetragonal LiYF4:Yb/Er. More importantly, at an ultra-low concentration of 0.5 mol% Li+ doping, as high as 34 times green and 101 times red emission enhancements are achieved.


Fiber intake and risk of colorectal cancer.
The relationship between various types of fiber and colorectal cancer risk was investigated using data from a case-control study conducted between January 1992 and June 1996 in Italy. The study included 1953 cases of incident, histologically confirmed colorectal cancers (1225 colon cancers and 728 rectal cancers) admitted to the major teaching and general hospitals in the study areas and 4154 controls with no history of cancer admitted to hospitals in the same catchment areas for acute nonneoplastic diseases. Dietary habits were investigated using a validated food frequency questionnaire. Odds ratios (ORs) were computed after allowance for age, sex, and other potential confounding factors, including physical activity and protein, fat, and carbohydrate intake. Fiber was analyzed both as a continuous variable and in quintiles. For most types of fiber, the OR of colon and rectal cancers was significantly below 1, and no appreciable differences emerged between the two. When the unit was set at the difference between the upper cutpoints of the fourth and first quintile, i.e., the 80th and 20th percentiles, the ORs for colorectal cancer were 0.68 for total fiber (determined by the Englyst method as nonstarch polysaccharides), 0.67 for soluble noncellulose polysaccharides (NCPs), 0.71 for total insoluble fiber, 0.67 for cellulose, 0.82 for insoluble NCPs, and 0.88 for lignin. When fiber was classified according to the source, the OR was 0.75 for vegetable fiber, 0.85 for fruit fiber, and 1.09 for cereal fiber. The ORs were similar for the two sexes and the strata of age, education, physical activity, family history of colorectal cancer, and energy intake. Likewise, no appreciable differences emerged when subsites of the colon and rectum were investigated separately. This study provides additional support for a protective and independent effect of fiber on colorectal cancer, particularly for cellulose and soluble NCPs, and of fiber of vegetable or fruit origin.


Omeprazole promotes proximal duodenal mucosal bicarbonate secretion in humans.
The proton pump inhibitor, omeprazole, surprisingly resulted in higher rates of proximal duodenal mucosal bicarbonate secretion than previously reported using an H2 receptor antagonist for gastric acid inhibition. Gastroduodenal perfusions were performed in healthy volunteers to evaluate whether this incidental finding is explained by more potent gastric acid inhibition by omeprazole or might be caused by the different mode of drug action. Basal and stimulated gastric and duodenal bicarbonate secretion rates were measured in the same subjects in control experiments (n = 17) and after pretreatment with high dose omeprazole (n = 17) and ranitidine (n = 9), respectively, by use of a technique permitting simultaneous measurements. Concentrations of bicarbonate were measured in the respective effluents by the method of back titration. Both omeprazole and ranitidine completely inhibited gastric acid secretion (pH 6.9 v 6.8; p > 0.05). Omeprazole caused higher rates of basal (mean (SEM)) (597 (48) v 351 (39) mumol/h; p < 0.02) and vagally stimulated (834 (72) v 474 (66) mumol/h; p < 0.02), but not acid stimulated (3351 (678) v 2550 (456) mumol/h; p > 0.05) duodenal bicarbonate secretion compared with control experiments. Also the combination of omeprazole and ranitidine increased (p = 0.05) duodenal bicarbonate secretion, while ranitidine alone caused no change in either basal or stimulated secretion. In the stomach basal as well as vagally stimulated bicarbonate secretion was independent of the means of acid inhibition. These results show that the proton pump inhibitor, omeprazole, promotes proximal duodenal mucosal bicarbonate secretion apparently independent of its gastric acid inhibitory effect. The mechanism of action remains speculative.


The Norfolk Island Eye Study (NIES): rationale, methodology and distribution of ocular biometry (biometry of the bounty).
To describe the recruitment, ophthalmic examination methods and distribution of ocular biometry of participants in the Norfolk Island Eye Study, who were individuals descended from the English Bounty mutineers and their Polynesian wives. All 1,275 permanent residents of Norfolk Island aged over 15 years were invited to participate, including 602 individuals involved in a 2001 cardiovascular disease study. Participants completed a detailed questionnaire and underwent a comprehensive eye assessment including stereo disc and retinal photography, ocular coherence topography and conjunctival autofluorescence assessment. Additionally, blood or saliva was taken for DNA testing. 781 participants aged over 15 years were seen (54% female), comprising 61% of the permanent Island population. 343 people (43.9%) could trace their family history to the Pitcairn Islanders (Norfolk Island Pitcairn Pedigree). Mean anterior chamber depth was 3.32mm, mean axial length (AL) was 23.5mm, and mean central corneal thickness was 546 microns. There were no statistically significant differences in these characteristics between persons with and without Pitcairn Island ancestry. Mean intra-ocular pressure was lower in people with Pitcairn Island ancestry: 15.89mmHg compared to those without Pitcairn Island ancestry 16.49mmHg (P = .007). The mean keratometry value was lower in people with Pitcairn Island ancestry (43.22 vs. 43.52, P = .007). The corneas were flatter in people of Pitcairn ancestry but there was no corresponding difference in AL or refraction. Our study population is highly representative of the permanent population of Norfolk Island. Ocular biometry was similar to that of other white populations. Heritability estimates, linkage analysis and genome-wide studies will further elucidate the genetic determinants of chronic ocular diseases in this genetic isolate.


[Transurethral microwave thermotherapy for benign prostatic hypertrophy in patients with high risk for surgery].
The effectiveness of transurethral microwave thermotherapy (TUMT) for benign prostatic hypertrophy in poor surgical risk patients (ASA class IV) with indwelling catheters, was assessed. All had had an indwelling catheter for 1-12 months. Removal of the catheter was possible in 14 out of the 24 (58.3%). Urinary peak flow rates were 12.2 +/- 3.5 ml/sec at 3 months of follow-up and post-voiding residual urine volumes of less than 50 ml were recorded in 13 catheter-free patients. Our data suggest that TUMT is an effective procedure for management of high risk patients with indwelling catheters in whom surgery or anesthesia are contraindicated.


Cost-effectiveness of bivalirudin versus heparin plus glycoprotein IIb/IIIa inhibitor in the treatment of non-ST-segment elevation acute coronary syndromes.
This study sought to assess the cost-effectiveness of bivalirudin versus heparin plus glycoprotein IIb/IIIa inhibitor (GPI) in thienopyridine-treated non-ST-segment elevation acute coronary syndrome (NSTE-ACS) patients undergoing early or urgent invasive management, from a United Kingdom National Health Service perspective. A decision-analytic model with lifelong time horizon was populated with event risks and resource use parameters derived from the Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) trial raw data. In a parallel analysis, key comparator strategy inputs came from Global Registry of Acute Coronary Events (GRACE) patients enrolled in the United Kingdom. Upstream and catheter laboratory-initiated GPI were assumed to be tirofiban and abciximab, respectively. Life expectancy of first-year survivors, unit costs, and health-state utilities came from United Kingdom sources. Costs and effects were discounted at 3.5%. Incremental cost-effectiveness ratios (ICERs) were expressed as cost per quality-adjusted life year (QALY) gained. Higher acquisition costs for bivalirudin were partially offset by lower hospitalization and bleeding costs. In the ACUITY-based analysis, per-patient lifetime costs in the bivalirudin and heparin plus GPI strategies were £10,903 and £10,653, respectively. Patients survived 10.87 and 10.82 years on average, corresponding to 5.96 and 5.93 QALYs and resulting in an ICER of £9,906 per QALY gained. The GRACE-based ICER was £12,276 per QALY gained. In probabilistic sensitivity analysis, 72.1% and 67.0% of simulation results were more cost-effective than £20,000 per QALY gained, in the ACUITY-based and GRACE-based analyses, respectively. Additional scenario analyses implied that greater cost-effectiveness may be achieved in actual clinical practice. Treating NSTE-ACS patients undergoing invasive management with bivalirudin is likely to represent a cost-effective option for the United Kingdom, when compared with the current practice of using heparin and a GPI.


Density and length in the neighborhood: Explaining cross-linguistic differences in learning to read in English and Dutch.
Two experiments examined underlying cognitive processes that may explain why it is harder to learn to read in English than in more transparent orthographies such as German and Dutch. Participants were English and Dutch readers from Grades 3 and 4. Experiment 1 probed the transition from serial to more parallel processing, as measured by the word length effect for words and pseudowords. English children took longer to make the transition to more parallel reading strategies for words than Dutch children. In contrast, Dutch children continued to use more serial reading strategies for pseudowords. Experiment 2 investigated children's sensitivity to the orthographic overlap between words, as measured by the size of orthographic neighborhood effects for words and pseudowords. Children reading Dutch showed greater sensitivity to the overlap between both words and pseudowords than English children. Cross-linguistic differences in the transition from serial to parallel reading strategies are discussed within the framework offered by the self-teaching hypothesis and the orthographic depth hypothesis. Finally, it is argued that differences between the two languages in the effect of orthographic neighborhood size are a result of cross-linguistic differences in orthographic density and not cross-linguistic differences in orthographic transparency.


Effect of green tea supplementation on the microbiological, antioxidant, and sensory properties of probiotic milks.
Green tea and its constituents are known for a wide range of health-promoting properties. They may exert antimicrobial action but without altering lactic acid bacteria. The aim of the present study was to estimate the effect of green tea addition on the selected properties of probiotic milks. Bioyogurts (fermented with ABT-1 coculture of Streptococcus thermophilus, Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12) and acidophilus milks (fermented with pure L. acidophilus LA-5 culture) with addition of 0, 5, 10, or 15% (v/v) green tea infusion (GTI) were produced and analyzed for the antioxidant capacity by the "diphenyl picrylhydrazyl" (DPPH) and "ferric-reducing antioxidant power" (FRAP) methods, acidity, the count of starter bacteria, and sensory properties at the 1st, 7th, 14th, and 21st day of cold storage. The 15% addition of GTI to the acidophilus milk significantly reduced the lactic acid production during the whole study. The GTI had no impact on the level of S. thermophilus and B. lactis BB-12 in bioyogurts, and its effect on the count of L. acidophilus LA-5 depended on the concentration and probiotic milk type. GTI similarly and in a dose-dependent manner enhanced the antioxidant capacity of both milk types. There were no significant differences between the sensory notes received for bioyogurts, whereas acidophilus milks with tea were less appreciated by the panelists. In conclusion, green tea could be successfully used as a functional additive for selected probiotic milks enhancing their health benefits, but the proper selection of tea additive and starter culture is recommended.


The influence of maximum safe resection of glioblastoma on survival in 1229 patients: Can we do better than gross-total resection?
Glioblastoma multiforme (GBM) is the most common and deadliest primary brain tumor. The value of extent of resection (EOR) in improving survival in patients with GBM has been repeatedly confirmed, with more extensive resections providing added advantages. The authors reviewed the survival of patients with significant EORs and assessed the relative benefit/risk of resecting 100% of the MRI region showing contrast-enhancement with or without additional resection of the surrounding FLAIR abnormality region, and they assessed the relative benefit/risk of performing this additional resection. The study cohort included 1229 patients with histologically verified GBM in whom ≥ 78% resection was achieved at The University of Texas MD Anderson Cancer Center between June 1993 and December 2012. Patients with > 1 tumor and those 80 years old or older were excluded. The survival of patients having 100% removal of the contrast-enhancing tumor, with or without additional resection of the surrounding FLAIR abnormality region, was compared with that of patients undergoing 78% to < 100% EOR of the enhancing mass. Within the first subgroup, the survival durations of patients with and without resection of the surrounding FLAIR abnormality were subsequently compared. The data on patients and their tumor characteristics were collected prospectively. The incidence of 30-day postoperative complications (overall and neurological) was noted. Complete resection of the T1 contrast-enhancing tumor volume was achieved in 876 patients (71%). The median survival time for these patients (15.2 months) was significantly longer than that for patients undergoing less than complete resection (9.8 months; p < 0.001). This survival advantage was achieved without an increase in the risk of overall or neurological postoperative deficits and after correcting for established prognostic factors including age, Karnofsky Performance Scale score, preoperative contrast-enhancing tumor volume, presence of cyst, and prior treatment status (HR 1.53, 95% CI 1.33-1.77, p < 0.001). The effect remained essentially unchanged when data from previously treated and previously untreated groups of patients were analyzed separately. Additional analyses showed that the resection of ≥ 53.21% of the surrounding FLAIR abnormality beyond the 100% contrast-enhancing resection was associated with a significant prolongation of survival compared with that following less extensive resections (median survival times 20.7 and 15.5 months, respectively; p < 0.001). In the multivariate analysis, the previously treated group with < 53.21% resection had significantly shorter survival than the 3 other groups (that is, previously treated patients who underwent FLAIR resection ≥ 53.21%, previously untreated patients who underwent FLAIR resection < 53.21%, and previously untreated patients who underwent FLAIR resection ≥ 53.21%); the previously untreated group with ≥ 53.21% resection had the longest survival. What is believed to be the largest single-center series of GBM patients with extensive tumor resections, this study supports the established association between EOR and survival and presents additional data that pushing the boundary of a conventional 100% resection by the additional removal of a significant portion of the FLAIR abnormality region, when safely feasible, may result in the prolongation of survival without significant increases in overall or neurological postoperative morbidity. Additional supportive evidence is warranted.


Evaluation of myofibroblasts by expression of alpha smooth muscle actin: a marker in fibrosis, dysplasia and carcinoma.
Evaluation of Myofibroblasts by studying expression of Alpha smooth muscle actin: A marker of Fibrosis, Dysplasia and Carcinoma. Myofibroblasts are cells that have contractile properties and are involved in inflammation, wound healing, fibrosis and oncogenesis in most of the organs and tissues. They are involved in healing and granulation tissue formation which occur after tissue injuries, also produce inflammatory mediators, growth factors and help in extracellular matrix reorganization by secretion of proteins like collagen, fibronectin, etc. Because of their component, Alpha smooth muscle actin ([alpha]-SMA), they are involved in the contraction of extracellular matrix and aid in tissue contraction. The myofibroblasts disappear by apoptosis after completion of repair, but their persistence causes a dysfunction in the repair mechanism, leading to excessive contraction and extracellular matrix (ECM) secretion and thus, fibrosis. The purpose of this study was to evaluate the presence of myofibroblasts in cases of Oral Submucous fibrosis (OSMF), which consisted of very early, early and moderately advanced OSMF, OSMF with dysplasia and oral squamous cell carcinoma (OSCC), by detecting (alpha)-SMA, which is a specific marker for myofibroblasts. The study sample consisted of three groups which comprised of 41 cases of OSMF, 10 cases of OSMF with dysplasia and 11 cases of OSCC. All the cases were subjected to immunohistochemistry by using (alpha)-SMA antibody for detection of myofibroblasts. The presence of myofibroblasts was significantly higher in oral squamous cell carcinomas as compared to that in OSMF with dysplasia and OSMF. A statistical significance was also noted between the staining index and age of the individuals and the staining index and duration of the habit. Myofibroblasts play a role in fibrosis, as was seen in OSMF. Activated myofibroblasts secrete proteolytic enzymes and cause matrix degradation, which is instrumental in cancer cell invasion and metastasis. Further studies in which the myofibroblasts are targetted, may help in providing therapeutic regimens in fibrosis, dysplasia and cancer.


Cytotoxic and anticancer properties of the Malaysian mangrove pit viper (Trimeresurus purpureomaculatus) venom and its disintegrin (purpureomaculin).
The Asiatic pit vipers from the Trimeresurus complex are medically important venomous snakes. These pit vipers are often associated with snakebite that leads to fatal coagulopathy and tissue necrosis. The cytotoxic venoms of Trimeresurus spp.; however, hold great potential for the development of peptide-based anticancer drugs. This study investigated the cytotoxic effect of the venom from Trimeresurus purpureomaculatus, the mangrove pit viper (also known as shore pit viper) which is native in Malaysia, across a panel of human cancer cell lines from breast, lung, colon and prostate as well as the corresponding normal cell lines of each tissue. The venom exhibited dose-dependent cytotoxic activities on all cell lines tested, with median inhibition concentrations (IC50) ranging from 0.42 to 6.98 µg/mL. The venom has a high selectivity index (SI = 14.54) on breast cancer cell line (MCF7), indicating that it is significantly more cytotoxic toward the cancer than to normal cell lines. Furthermore, the venom was fractionated using C18 reversed-phase high-performance liquid chromatography and the anticancer effect of each protein fraction was examined. Fraction 1 that contains a hydrophilic low molecular weight (approximately 7.5 kDa) protein was found to be the most cytotoxic and selective toward the breast cancer cell line (MCF7). The protein was identified using liquid chromatography-tandem mass spectrometry as a venom disintegrin, termed purpureomaculin in this study. Taken together, the findings revealed the potent and selective cytotoxicity of a disintegrin protein isolated from the Malaysian T. purpureomaculatus venom and suggested its anticancer potential in drug discovery.


Minimum 5-Year Outcomes of Robotic-assisted Primary Total Hip Arthroplasty With a Nested Comparison Against Manual Primary Total Hip Arthroplasty: A Propensity Score-Matched Study.
Robotic-assisted technology has been a reliable tool in enhancing precision and accuracy of cup placement in total hip arthroplasty (THA). Still, questions remain on the clinical benefit of this technology. The purposes of the ongoing study were (1) to report on minimum 5-year outcomes in patients who underwent robotic-assisted primary THA (rTHA), (2) to compare those outcomes to a propensity score-matched manual primary THA (mTHA) control group, and (3) to compare radiographic measures between the groups regarding acetabular cup placement. Prospectively collected patient data were retrospectively reviewed for primary THA recipients during June 2008 to July 2013. Patients with minimum 5-year follow-up for Harris Hip Score, Forgotten Joint Score-12, Veterans RAND-12 Mental, Veterans RAND-12 Physical, 12-Item Short Form Survey Mental, 12-Item Short Form Survey Physical, visual analog scale, and satisfaction were included. Patient-reported outcomes, cup placement, and revision rate of the rTHA group were compared with those of a propensity score-matched mTHA control group. Sixty-six rTHAs were matched to 66 mTHAs. The rTHA group reported significantly higher Harris Hip Score, Forgotten Joint Score-12, Veterans RAND-12 Physical, and 12-Item Short Form Survey Physical (P < 0.001, P = 0.002, P = 0.002, P = 0.001). The acetabular implant placement by rTHA had a 9 and 4.7-fold reduced risk of placement outside the Lewinnek and Callanan safe zones, respectively (relative risk, 0.11 [95% confidence interval, 0.03 to 0.46]; P = 0.002; relative risk, 0.21 [95% confidence interval, 0.01 to 0.47]; P = 0.001). In addition, rTHA recipients had lesser absolute values of leg length discrepancy and global offset (P = 0.091, P = 0.001). Patients who received rTHA reported favorable outcomes at minimum 5-year follow-up. Furthermore, in comparison to a propensity score pair-matched mTHA group, rTHAs reported higher patient-reported outcome scores and had 89% reduced risk of acetabular implant placement beyond the Lewinnek safe zone and 79% reduced risk of placement beyond the Callanan safe zone. Level III.


Fr-TM-align: a new protein structural alignment method based on fragment alignments and the TM-score.
Protein tertiary structure comparisons are employed in various fields of contemporary structural biology. Most structure comparison methods involve generation of an initial seed alignment, which is extended and/or refined to provide the best structural superposition between a pair of protein structures as assessed by a structure comparison metric. One such metric, the TM-score, was recently introduced to provide a combined structure quality measure of the coordinate root mean square deviation between a pair of structures and coverage. Using the TM-score, the TM-align structure alignment algorithm was developed that was often found to have better accuracy and coverage than the most commonly used structural alignment programs; however, there were a number of situations when this was not true. To further improve structure alignment quality, the Fr-TM-align algorithm has been developed where aligned fragment pairs are used to generate the initial seed alignments that are then refined using dynamic programming to maximize the TM-score. For the assessment of the structural alignment quality from Fr-TM-align in comparison to other programs such as CE and TM-align, we examined various alignment quality assessment scores such as PSI and TM-score. The assessment showed that the structural alignment quality from Fr-TM-align is better in comparison to both CE and TM-align. On average, the structural alignments generated using Fr-TM-align have a higher TM-score (~9%) and coverage (~7%) in comparison to those generated by TM-align. Fr-TM-align uses an exhaustive procedure to generate initial seed alignments. Hence, the algorithm is computationally more expensive than TM-align. Fr-TM-align, a new algorithm that employs fragment alignment and assembly provides better structural alignments in comparison to TM-align. The source code and executables of Fr-TM-align are freely downloadable at: http://cssb.biology.gatech.edu/skolnick/files/FrTMalign/.


Synthesis of a poly(L-lysine)-calcium phosphate hybrid on titanium surfaces for enhanced bioactivity.
Titanium has been a successful implant material owing to its excellent strength to weight ratio, toughness, and bioinert oxide surface. Significant progress has been made on the improvement of titanium's bioactivity by coating its oxide surface with calcium phosphates and bioactive molecules. Here, we report on the coating of titanium with a poly(L-lysine)-calcium phosphate hybrid material with a nanoscale texture. This hybrid coating was grown by first nucleating seed crystals of calcium phosphate, directly on the Ti surface and then exposing this surface to solutions containing Ca(2+), PO(4)(3-), and poly(L-lysine). The resultant hybrid coating was characterized by electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray photoelectron spectroscopy, and elemental analysis. This material contained 14% by weight poly(L-lysine), and this organic component decreased greatly the dimensions of the surface features, thus enhancing surface area relative to the inorganic control. The highly textured hybrid material was more susceptible than the control to acidic and enzymatic degradation. The amino acid cysteine was covalently linked to the hybrid material, demonstrating the potential of this coating for further functionalization. These hybrid coatings may prove useful in enhancing the bioactivity of titanium.