Evaluation of glucose-6-phosphate dehydrogenase serum level in patients with multiple sclerosis and neuromyelitis optica.
Background: Multiple sclerosis (MS) and neuromyelitis optica (NMO) are both demyelinating disorders and oxidative stress is suggested to have a role in their pathogenesis. Glucose-6-phosphate dehydrogenase (G6PD) produces nicotinamide adenine dinucleotide phosphate (NADPH) via the pentose phosphate pathway. NADPH is not only involved in the synthesis of fatty acids necessary for myelination, but also it is involved in the defense against oxidative stress. Prescribing supplementary vitamin D as a part of the MS treatment plan can increase G6PD gene expression. The aim of this study was to determine the serum level of G6PD in patients with MS and NMO and its relationship with vitamin D, since it is yet to be explored thoroughly. Methods: In this case-control study, subjects were divided into three experimental and control groups. The experimental groups comprised 50 patients with relapsing-remitting MS (RRMS) who had a history of vitamin D consumption, 50 newly-diagnosed MS patients, and 50 patients with NMO. Control group included 65 healthy individuals. Serum level of G6PD was measured and compared among these groups. Results: No significant difference was seen between the G6PD level in patients with MS and NMO, but it should be noted that this level was significantly lower than the healthy group. G6PD serum level was significantly higher in patients with MS who had previously consumed supplementary vitamin D compared to those who had not. Conclusion: G6PD deficiency is observed in patients with MS and NMO. Also, supplementary vitamin D may induce favorable results on the G6PD level.


Feeders of Free-Roaming Cats: Personal Characteristics, Feeding Practices, and Data on Cat Health and Welfare in an Urban Setting of Israel.
Cat feeders serve as an important source of available food for free-roaming cats (FRCs) and can play a central role in providing data on FRC distribution, welfare, and health. Data on cat feeder personalities as well as a better understanding of their feeding practices offer relevance for decision making concerning FRC population control strategies. The current study surveyed 222 FRC feeders who responded to a municipal trap-neuter-return (TNR) campaign in an Israeli central urban setting. The aim of the study was to describe their personal characteristics, feeding practices, and the FRC populations they feed. Feeders were divided into four groups according to the number of cats they claimed to feed per day (group 1: fed up to 5 cats, group 2: fed 6-10 cats, group 3: fed 11-20 cats, and group 4: fed ≥21 cats). Most feeders were women (81%), with a median age of 58 years (range 18-81). The feeders reported an overall feeding of 3337 cats in 342 different feeding locations. Feeders of group 4 comprised 15.31% (n = 34) of all feeders but fed 56% (n = 1869) of the FRC in 37.42% (n = 128) of the feeding locations. "Heavy" feeders (groups 3 and 4) reported that they traveled significantly longer distances in order to feed the cats. Commercial dry food consisted of 90% of the food they provided, with 66% of them feeding once a day, with less food per cat per day than the other feeder groups. Interestingly, "heavy" feeders were usually singles, had on average fewer siblings, a clear preference for owning cats as pets, and lived in lower income neighborhoods. According to the feeders' reports on the FRC populations they fed, 69.7% (2325/3337) cats were neutered and 11.8% (395/3337) were kittens. In addition, they reported that 1.6% (54/3337) of the cats were limping, 2% (67/3337) suffered from a systemic disease, 4% (135/3337) had skin lesions, and 3.9% (130/3337) were suffering from a chronic disability. Abundance of kittens and morbidity rate were significantly and negatively associated with neutering rate. These findings are in accordance with the suggestion that neutering may potentially improve cat welfare by reducing morbidity. Collaboration by the authorities with these heavy feeders, who represent a small number of FRC feeders and feed substantial FRC numbers, may be significant for the control and monitoring of FRC populations and their resources.


Community-Based Pharmacists' Needs for HIV-Related Training and Experience.
To examine pharmacists' self-reported competence in providing care to persons living with HIV (PLWH) and their HIV-related training and experience needs. We interviewed 28 community-based pharmacists providing care to PLWH in 4 Midwestern cities. Less than half (46%) of the pharmacists considered themselves competent to provide PLWH care, and less than a third (29%) worked with PLWH during their pharmacy residency. Specialty pharmacists need training on opportunistic infections and HIV-related comorbidities, nonspecialty pharmacists need general training in HIV treatment and patient communications skills, and all pharmacists require a mechanism to keep updated in the latest HIV treatment recommendations. In the current era of patient-centered care, a pharmacist that is well rounded-not just in dispensing antiretroviral medications but highly knowledgeable in different aspects of ART and HIV-specific patient care-can make a valuable contribution to the health care team. Pharmacy school curricula and continuing professional education need to be aligned to meet the knowledge and competency needs of community pharmacists who are strategically positioned to provide care to PLWH.


Contrast-enhanced CT quantification of the hepatic fractional extracellular space: correlation with diffuse liver disease severity.
The purpose of this study was to determine whether contrast-enhanced CT quantification of the hepatic fractional extracellular space (ECS) correlates with the severity of diffuse liver disease. The cases of 70 patients without (46 men, 24 women; mean age, 59.1 years) and 36 patients with (23 men, 13 women; mean age, 63.1 years) cirrhosis who had undergone unenhanced and 10-minute delayed phase contrast-enhanced CT were retrospectively identified. By consensus one experienced radiologist and one trainee measured the CT attenuation of the liver and aorta to estimate the fractional ECS, defined as the ratio of the difference between the attenuation of the liver on 10-minute and unenhanced images to the difference between the attenuation of the aorta on 10-minute and unenhanced images multiplied by 1 minus the hematocrit. Findings were correlated with each patient's Model of End-Stage Liver Disease (MELD) score. The mean MELD score was higher in patients with than in those without cirrhosis (14.3 ± 7.3 versus 7.20 ± 2.4, p < 0.0001). The mean fractional ECS was significantly greater in patients with cirrhosis than in those without cirrhosis (41.0% ± 9.0% versus 23.8% ± 6.3%, p < 0.0001). The fractional ECS correlated with the MELD score (r = 0.572, p < 0.0001) and was predictive of cirrhosis with an area under the receiver operating characteristic curve of 0.953 (p < 0.0001). The sensitivity and specificity of an expanded fractional ECS greater than 30% for the prediction of cirrhosis were 92% and 83%. Multivariate linear regression revealed that the fractional ECS is complementary to the MELD score as a predictor of cirrhosis (p < 0.0001). Noninvasive contrast-enhanced CT quantification of the fractional ECS correlates with the MELD score, an indicator of the severity of liver disease, and merits further study.


Dealing with the long-term social implications of research.
Biomedical and behavioral research may affect strongly held social values and thereby create significant controversy over whether such research should be permitted in the first place. Institutional review boards (IRBs) responsible for protecting the rights and welfare of participants in research are sometimes faced with review of protocols that have significant implications for social policy and the potential for negative social consequences. Although IRB members often raise concerns about potential long-term social implications in protocol review, federal regulations strongly discourage IRBs from considering them in their decisions. Yet IRBs often do consider the social implications of research protocols and sometimes create significant delays in initiating or even prevent such research. The social implications of research are important topics for public scrutiny and professional discussion. This article examines the reasons that the federal regulations preclude IRBs from assessing the social risks of research, and examines alternative approaches that have been used with varying success by national advisory groups to provide such guidance. The article concludes with recommendations for characteristics of a national advisory group that could successfully fulfill this need, including sustainability, independence, diverse and relevant expertise, and public transparency.


Connection and recovery in the COVID-19 age: An analysis of changes in goal-setting throughout the pandemic by consumers living with enduring mental illness.
Goal-setting is a tool that empowers consumer recovery. Though the pandemic has affected consumer goal-setting, the nature and extent of this impact have not been examined in a recovery setting. The aim of this study is to assess whether the recovery goals of individuals with serious mental illness changed in association with the COVID-19 pandemic. In this mixed-methods design, data were collected from a purposeful sample of consumers (nTOTAL = 355) aged 19-67 years (MAGE = 44.56, SD = 13.05) attending Recovery Camp, a 5-day therapeutic-recreation programme for individuals living with severe mental illness (e.g., PTSD, schizophrenia). Consumer-set goals were examined across 5 programmes prior to March 2020 (nPRE = 126) and 11 following (nPOST = 229). Goals were set on day one, with attainment self-scored on day five. Chi-squared goodness-of-fit tests compared goal proportions per domain; tests of independence assessed changes in goals pre- and post-pandemic. Six goal domains were identified: Approach-Based Recovery, Avoidance-based Recovery, Novel Physical Activities, Relationships, Health, and Recreation/Relaxation. Irrespective of the pandemic, goal attainment was consistently high across all programmes (86.56%). Approach-based Recovery goals were predominant pre-pandemic, but were significantly reduced post-pandemic (p = 0.040). Goals related to Relationships and Novel Physical Activities took precedence throughout the pandemic. Post-COVID-19, consumer recovery goals reveal increased desire for connection, novelty-seeking, and positive behavioural change.


Temperature-sensitive initiation and elongation of adenovirus DNA replication in vitro with nuclear extracts from H5ts36-, H5ts149-, and H5ts125-infected HeLa cells.
Adenovirus DNA replication was studied in vitro in nuclear extracts prepared from HeLa cells infected at the permissive temperature with H5ts125, H5ts36, or H5ts149, three DNA-negative mutants belonging to two different complementation groups. At the restrictive temperature, H5ts125 extracts, containing a thermolabile 72-kilodalton DNA-binding protein, enable the formation of an initiation complex between the 82-kilodalton terminal protein precursor (pTP) and dCTP, but further elongation of this complex is inhibited. Wild-type DNA-binding protein or a 47-kilodalton chymotryptic DNA-binding fragment can complement the mutant protein in the elongation reaction. No difference in heat inactivation was observed between wild-type extracts and H5ts36 or H5ts149 extracts when the replication of terminal XbaI fragments of adenovirus type 5 DNA-terminal protein complex was studied. In contrast, the formation of a pTP-dCMP initiation complex, as well as the partial elongation reaction up to nucleotide 26, were consistently more temperature sensitive in mutant extracts. The results suggest that the H5ts36/H5ts149 gene product is required for initiation of adenovirus type 5 DNA replication and that the 72-kilodalton DNA-binding protein functions early in elongation.


A quantitative trait locus genome scan for porcine muscle fiber traits reveals overdominance and epistasis.
Muscle histochemical characteristics are decisive determinants of meat quality. The relative percentage and diameters of the different muscular fiber types influence crucial aspects of meat such as color, tenderness, and ultimate pH. Despite its relevance, however, the information on muscle fiber genetic architecture is scant, because histochemical muscle characterization is a laborious task. Here we report a complete QTL scan of muscle fiber traits in 160 animals from a F(2) cross between Iberian and Landrace pigs using 139 markers. We identified 20 genome regions distributed along 15 porcine chromosomes (SSC1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and X) with direct and(or) epistatic effects. Epistasis was frequent and some interactions were highly significant. Chromosomes 10 and 11 seemed to behave as hubs; they harbored 2 individual QTL, but also 6 epistatic regions. Numerous individual QTL effects had cryptic alleles, with opposite effects to phenotypic pure breed differences. Many of the QTL identified here coincided with previous reports for these traits in the literature, and there was overlapping with potential candidate genes and previously reported meat quality QTL.


Amplified RNase H activity in Escherichia coli B/r increases sensitivity to ultraviolet radiation.
Strains of E. coli B/r transformed with the plasmid pSK760 were found to be sensitized to inactivation by ultraviolet radiation (UV) and to have elevated levels of RNase H activity. Strains transformed with the carrier vector pBR322 or the plasmid pSK762C derived from pSK760 but with an inactivated rnh gene were not sensitized. UV-inactivation data for strains having known defects in DNA repair and transformed with pSK760 suggested an interference by RNase H of postreplication repair: uvrA cells were strongly sensitized, wild-type and uvrA recF cells were moderately sensitized and recA cells were not sensitized; and minimal medium recovery was no longer apparent in sensitized uvrA cells. Biochemical studies showed that post-UV DNA synthesis was sensitized and that the smaller amounts of DNA synthesized after irradiation, while of normal reduced size as indicated by sedimentation position in alkaline sucrose gradients, did not shift to a larger size (more rapidly sedimenting) upon additional incubation. We suggest an excess level of RNase H interferes with reinitiation of DNA synthesis on damaged templates to disturb the normal pattern of daughter strand gaps and thereby to inhibit postreplication repair.


The influence of depth and a subsea pipeline on fish assemblages and commercially fished species.
Knowledge of marine ecosystems that grow and reside on and around subsea oil and gas infrastructure is required to understand impacts of this offshore industry on the marine environment and inform decommissioning decisions. This study used baited remote underwater stereo-video systems (stereo-BRUVs) to compare species richness, fish abundance and size along 42.3 km of subsea pipeline and in adjacent areas of varying habitats. The pipeline is laid in an onshore-offshore direction enabling surveys to encompass a range of depths from 9 m nearshore out to 140 m depth offshore. Surveys off the pipeline were performed across this depth range and in an array of natural habitats (sand, macroalgae, coral reef) between 1 km and 40 km distance from the pipeline. A total of 14,953 fish were observed comprising 240 species (131 on the pipeline and 225 off-pipeline) and 59 families (39 on the pipeline and 56 off-pipeline) and the length of 8,610 fish were measured. The fish assemblage on and off the pipeline was similar in depths of <80 m. In depths beyond 80 m, the predominant habitat off-pipeline was sand and differences between fish assemblages on and off-pipeline were more pronounced. The pipeline was characterised by higher biomass and abundances of larger-bodied, commercially important species such as: Pristipomoides multidens (goldband snapper), Lutjanus malabaricus (saddletail snapper) and Lutjanus russellii (Moses' snapper) among others, and possessed a catch value 2-3 times higher per stereo-BRUV deployment than that of fish observed off-pipeline. Adjacent natural seabed habitats possessed higher abundances of Atule mate (yellowtail scad), Nemipterus spp. (threadfin bream) and Terapon jarbua (crescent grunter), species of no or low commercial value. This is the first published study to use stereo-BRUVs to report on the importance of subsea infrastructure to commercially important fishes over a depth gradient and increases our knowledge of the fish assemblage associated with subsea infrastructure off north-west Australia. These results provide a greater understanding of ecological and fisheries implications of decommissioning subsea infrastructure on the north-west shelf, and will help better inform decision-making on the fate of infrastructure at different depths.


Integrating solvation shell structure in experimentally driven molecular dynamics using x-ray solution scattering data.
In the past few decades, prediction of macromolecular structures beyond the native conformation has been aided by the development of molecular dynamics (MD) protocols aimed at exploration of the energetic landscape of proteins. Yet, the computed structures do not always agree with experimental observables, calling for further development of the MD strategies to bring the computations and experiments closer together. Here, we report a scalable, efficient MD simulation approach that incorporates an x-ray solution scattering signal as a driving force for the conformational search of stable structural configurations outside of the native basin. We further demonstrate the importance of inclusion of the hydration layer effect for a precise description of the processes involving large changes in the solvent exposed area, such as unfolding. Utilization of the graphics processing unit allows for an efficient all-atom calculation of scattering patterns on-the-fly, even for large biomolecules, resulting in a speed-up of the calculation of the associated driving force. The utility of the methodology is demonstrated on two model protein systems, the structural transition of lysine-, arginine-, ornithine-binding protein and the folding of deca-alanine. We discuss how the present approach will aid in the interpretation of dynamical scattering experiments on protein folding and association.


Identifying inflammatory bowel disease patients at risk of metabolic dysfunction-associated fatty liver disease: usefulness of non-invasive steatosis predictive scores.
Inflammatory bowel disease (IBD) patients have a higher risk of metabolic dysfunction-associated fatty liver disease (MAFLD) compared with the general population. However, it is not known whether available non-invasive hepatic steatosis scores are useful in predicting MAFLD in IBD patients. We aimed to analyze the performances of MAFLD screening score (MAFLD-S), Fatty Liver Index (FLI), Hepatic Steatosis Index (HSI) and Clinical Prediction Tool for NAFLD in Crohn's Disease (CPN-CD), in identifying MAFLD in IBD patients. A cross-sectional study was carried out including consecutive adult IBD outpatients submitted to transient elastography (TE). MAFLD criteria were assessed, and hepatic steatosis (HS) was defined by a controlled attenuation parameter (CAP) >248 dB/m using TE. MAFLD-S, FLI, HSI, and CPN-CD were calculated and their accuracy for the prediction of MAFLD was evaluated through their areas under the receiver-operating characteristic (AUROC) curves. Of 168 patients, body mass index ≥25, type 2 diabetes mellitus, dyslipidemia and arterial hypertension were present in 76 (45.2%), 10 (6.0%), 53 (31.5%), 20 (11.9%), respectively. HS was identified in 77 (45.8%) patients, of which 65 (84.4%) fulfilled MAFLD criteria. MAFLD-S (AUROC, 0.929 [95% CI, 0.888-0.971]) had outstanding and FLI (AUROC, 0.882 [95% CI, 0.830-0.934]), HSI (AUROC, 0.803 [95% CI, 0.736-0.871]), and CPN-CD (AUROC, 0.822 [95% CI, 0.753-0.890) had excellent discrimination in predicting MAFLD. MAFLD-S, FLI, HSI and CPN-CD scores can accurately identify MAFLD in IBD patients, allowing the selection of those in whom hepatic steatosis and metabolic risk factors assessment may be particularly beneficial.


[Prophylactic antibiotics in neurosurgery].
Because of a low risk of infection (around 2-3%), prophylactic use of antibiotics in neurosurgery is a controversial issue. Some neurosurgeons consider that there are strong arguments against the use of antimicrobials (promotion of antibiotic-resistant strains of bacteria, superinfection and adverse drug reactions) and meticulous aseptic techniques could be more usefully than prophylactic antibiotics. On the other hand, despite of being rare, the consequences of a neurosurgical infection can be dramatic and may result in a rapid death, caused by meningitis, cerebritis, abscess formation or sepsis. Clinical studies emphasized that the most important factors influencing the choice of antibiotic prophylaxis in neurosurgery is the patient's immune status, virulence of the pathogens and the type of surgery ("clean contaminated"--procedure that crosses the cranial sinuses, "clean non-implant"--procedure that does not cross the cranial sinuses, CSF shunt surgery, skull fracture). Prophylaxis has become the standard of care for contaminated and clean-contaminated surgery, also for surgery involving insertion of artificial devices. The antibiotic (first/second generation of cephalosporins or vancomycin in allergic patients) should recover only the cutaneous possibly contaminating flora (S. aureus, S. epidermidis) and should be administrated 30' before the surgical incision, intravenously in a single dose. Most studies pointed that identification of the risk factors for infections, correct asepsis and minimal prophylactic antibiotic regimen, help neurosurgeons to improve patient care and to decrease mortality without selecting resistant bacteria.


Inhibition of vesicular stomatitis virus RNA synthesis by 2',3'-dideoxycytidine 5'-triphosphate.
The ability of the compound 2',3'-dideoxycytidine 5'-triphosphate (ddCTP) to serve as an inhibitor of viral RNA synthesis was examined using an in vitro system that supports vesicular stomatitis virus (VSV) protein synthesis, transcription and replication. Viral RNA synthesis was inhibited by 87 and 98% of control, respectively, in reactions containing 1 mM- and 10 mM-ddCTP in place of CTP. VSV RNA replication and transcription were inhibited equally by ddCTP. At a concentration of 1 mM-ddCTP, there was no inhibitory effect on viral protein synthesis; at 10 mM-ddCTP, total protein synthesis was inhibited by 30% as compared to control reactions. The presence of ddCTP had no effect on the size or relative molar amounts of each protein synthesized as analysed by electrophoresis on polyacrylamide gels. This is the first report describing a compound that will inhibit VSV RNA synthesis in vitro without compromising the concurrent synthesis and modification of proteins.


Hepatitis C virus NS2 protein contributes to virus particle assembly via opposing epistatic interactions with the E1-E2 glycoprotein and NS3-NS4A enzyme complexes.
The hepatitis C virus NS2 protein has been recently implicated in virus particle assembly. To further understand the role of NS2 in this process, we conducted a reverse genetic analysis of NS2 in the context of a chimeric genotype 2a infectious cell culture system. Of 32 mutants tested, all were capable of RNA replication and 25 had moderate-to-severe defects in virus assembly. Through forward genetic selection for variants capable of virus spread, we identified second-site mutations in E1, E2, NS2, NS3, and NS4A that suppressed NS2 defects in assembly. Two suppressor mutations, E1 A78T and NS3 Q221L, were further characterized by additional genetic and biochemical experiments. Both mutations were shown to suppress other NS2 defects, often with mutual exclusivity. Thus, several NS2 mutants were enhanced by NS3 Q221L and inhibited by E1 A78T, while others were enhanced by E1 A78T and inhibited by NS3 Q221L. Furthermore, we show that the NS3 Q221L mutation lowers the affinity of native, full-length NS3-NS4A for functional RNA binding. These data reveal a complex network of interactions involving NS2 and other viral structural and nonstructural proteins during virus assembly.


British women's attitudes to surrogacy.
There has been little interest in the research literature on public opinions regarding assisted conception and surrogacy, particularly in European countries, despite the growing evidence showing that problems in adaptation and coping may be related to perceived normative values. This study investigated British women's attitudes to surrogacy using components of the theory of planned behaviour (TPB). Questionnaires on attitudes to surrogacy and reasons for parenthood were completed by 187 women from the general public. Significant socio-demographic differences were found between women who were possibly willing (n = 76) and those who were unwilling (n = 111) to become surrogate mothers. General attitudes to surrogacy also differed between groups (P = 0.000). This study supported the predictive utility of components of the TPB, and differentiated adequately between groups on attitudes to recruitment for surrogacy (P = 0.000), the consequences of surrogacy (P = 0.000), factors that induce people to become surrogates (P = 0.000), social support (P = 0.000), having personal control (P = 0.002) and reasons for parenthood (P = 0.000). Age (P = 0.000), attitudes to advertising (P = 0.02) and the consequences of surrogacy (P = 0.05) predicted (un)willingness to become a potential surrogate mother. Further research is needed with larger sample sizes of potential surrogates to determine whether the predictive attitudes reported here translate to actual behaviours. The larger group which was not interested in considering becoming a surrogate scored significantly more negatively on all attitudes towards surrogacy. The negative attitudes reported by the 'unwilling to consider being a surrogate' group may reflect attitudes held by the majority of the population and are likely to be influenced by reports of stigma associated with surrogacy.


Brain PET Imaging of α7-nAChR with [18F]ASEM: Reproducibility, Occupancy, Receptor Density, and Changes in Schizophrenia.
The α7 nicotinic acetylcholine receptor increasingly has been implicated in normal brain physiology, as well as in neuropsychiatric disorders. The highly cortical distribution of α7 nicotinic acetylcholine receptor suggests a role in cognition. We expanded the first-in-human PET imaging of α7 nicotinic acetylcholine receptor with [18F]ASEM from 5 to 21 healthy nonsmoking volunteers and added a feasibility study in 6 male patients with schizophrenia. Study aims included: (1) confirmation of test-retest reproducibility of [18F]ASEM binding, (2) demonstration of specificity by competition with DMXB-A, an α7 nicotinic acetylcholine receptor partial agonist, (3) estimation of [18F]ASEM binding potentials and α7 nicotinic acetylcholine receptor density in vivo in humans, and (4) demonstrating the feasibility of studying α7 nicotinic acetylcholine receptor as a target for schizophrenia. Test-retest PET confirmed reproducibility (>90%) (variability ≤7%) of [18F]ASEM volume of distribution (VT) estimates in healthy volunteers. Repeated sessions of PET in 5 healthy subjects included baseline and effect of inhibition after oral administration of 150 mg DMXB-A. From reduction of binding potentials, we estimated the dose-dependent occupancy of α7 nicotinic acetylcholine receptor by DMXB-A at 17% to 49% for plasma concentrations at 60 to 200 nM DMXB-A. In agreement with evidence postmortem, α7 nicotinic acetylcholine receptor density averaged 0.67 to 0.82 nM and inhibitor affinity constant averaged 170 to 385 nM. Median VT in a feasibility study of 6 patients with schizophrenia was lower than in healthy volunteers in cingulate cortex, frontal cortex, and hippocampus (P = 0.02, corrected for multiple comparions, Mann-Whitney test). The current results confirm the reproducibility of [18F]ASEM VT estimates and the specificity of the tracer for α7 nicotinic acetylcholine receptor. Preliminary findings from our feasibility study of [18F]ASEM binding in patients with schizophrenia are suggestive and provide guidance for future studies with more subjects.


Deconstructing the third dimension: how 3D culture microenvironments alter cellular cues.
Much of our understanding of the biological mechanisms that underlie cellular functions, such as migration, differentiation and force-sensing has been garnered from studying cells cultured on two-dimensional (2D) glass or plastic surfaces. However, more recently the cell biology field has come to appreciate the dissimilarity between these flat surfaces and the topographically complex, three-dimensional (3D) extracellular environments in which cells routinely operate in vivo. This has spurred substantial efforts towards the development of in vitro 3D biomimetic environments and has encouraged much cross-disciplinary work among biologists, material scientists and tissue engineers. As we move towards more-physiological culture systems for studying fundamental cellular processes, it is crucial to define exactly which factors are operative in 3D microenvironments. Thus, the focus of this Commentary will be on identifying and describing the fundamental features of 3D cell culture systems that influence cell structure, adhesion, mechanotransduction and signaling in response to soluble factors, which - in turn - regulate overall cellular function in ways that depart dramatically from traditional 2D culture formats. Additionally, we will describe experimental scenarios in which 3D culture is particularly relevant, highlight recent advances in materials engineering for studying cell biology, and discuss examples where studying cells in a 3D context provided insights that would not have been observed in traditional 2D systems.


Epoxides of carcinogenic polycyclic hydrocarbons are frameshift mutagens.
K-region epoxides of the carcinogens benz[a] anthracene, dibenz[a,h]anthracene, and 7-methylbenz[a] anthracene are mutagenic in strains of Salmonella typhimurium designed to detect frameshift mutagens. Parent hydrocarbons, K-region diols and phenols and some other epoxides are inactive as mutagens in these tests. Polycyclic hydrocarbon epoxides, and other presumed proximal carcinogens, are discussed as examples of intercalating agents with reactive side chains. It has been shown previously that intercalating agents with reactive side chains are potent frameshift mutagens.


Allelic divergence at B alpha 1 pheromone receptor genes of Schizophyllum commune.
The multiallelic mating type locus B alpha 1 of Schizophyllum commune encodes a pheromone receptor and putative pheromone genes. A comparison of two alleles encoding receptors that share the same specificity B alpha 1 was performed using strains of different geographic origin. The amino acid sequence alignment revealed strong conservation of the largest part of the receptor. Only in the distal C-terminus major amino acid was divergence encountered. This C-terminal region of 117 of the total of 639 amino acids was shown to be unnecessary for function in vivo by transformation experiments.


Criteria Suggestive of Psychological Components of Itch and Soma-toform Itch: Study of a Large Sample of Patients with Chronic Pruritus.
While psychological factors are relevant in many patients with chronic pruritus, not all patients can be offered psychologic, psychosomatic or psychiatric consultation. The aim of this exploratory study was to identify criteria suggestive of psychological factors relevant for the etiology of chronic pruritus and of somatoform pruritus. Routine data from the database of the Center for Chronic Pruritus of the University Hospital Münster were used, including the Neuroderm Questionnaire, Dermatology Life Quality Index and Hospital Anxiety and Depression Scale. Chronic pruritus patients (n = 3,391) without a psychiatric diagnosis in their medical history were compared to the 331 chronic pruritus patients with diagnoses of "psychological factors associated with etiology and course of chronic pruritus" (ICD-10:F54) or "somatoform pruritus" (F45.8) confirmed by an expert. The latter reported more pruritus triggers, especially "strain" and "emotional tension" and used more emotional adjectives to describe their pruritus. They reported more often scratching leading to excoriations, higher levels of pruritus, impairment of quality of life, anxiety and depression. These aspects suggest the presence of psychological factors in the etiology of chronic pruritus and somatoform pruritus. Prospective validation, however, needs to be carried out.


Ileus in the critically ill: causes, treatment and prevention.
Bowel dysfunction, especially ileus, has been increasingly recognized in critically ill patients. Ileus is commonly associated to constipation, however abnormal motility can also concern the upper digestive tract, therefore impaired gastrointestinal transit (IGT) seems to be a more appropriate term. IGT, especially constipation, is common among patients under mechanical ventilation, occurring in up to 80% of the patients during the first week, and has been associated with worse outcome in intensive care unit (ICU). It is acknowledged that the most relevant definition for constipation in ICU is the absence of stool for the first six days after admission. Concerning the upper digestive intolerance (UDI), the diagnosis should rely only on vomiting and the systematic gastric residual volume (GRV) monitoring should be avoided. IGT results from a complex pathophysiology in which both the critical illness and its specific treatments may have a deleterious role. Both observational and experimental studies have shown the deleterious effect of sepsis, multiorgan failure, sedation (especially opioids) and mechanical ventilation on gut function. To date few studies have reported effect of treatment on IGT and the level of evidence is low. However, cholinesterase inhibitors seem safe and could probably be used in case of constipation but remains poorly prescribed. Prevention with bowel management protocol using osmotic laxatives appears to be safe but did not demonstrate its effectiveness. For patients treated with high posology of opioids during sedation, enteral opioid antagonists may be a promising strategy.


Two-dimensional electrophoresis and western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes.
The allergenicity of seed storage proteins, the major components of edible legume seeds, may cause serious reactions in both children and adult population. Updated methodologies for evaluation of the activity of these proteins are needed. In this paper we used two-dimensional (2D) electrophoretic techniques to investigate the immuno-cross-reactivities of anti Ara h 3 basic subunit IgG to the seed proteomes of three legume species, namely, peanut, soybean, and lupin. The seed proteins, extracted with two different procedures, were separated by 2D electrophoresis, and the electrophoretic maps were analyzed by Western blot. In peanut proteome the antibodies strongly reacted with the 23 kDa polypeptides, corresponding to Ara h 3 basic isoforms, the antigen they were raised to, and three unidentified acidic polypeptides near 45 kDa. Remarkable cross-reactivities with lupin and soybean Ara h 3 homologous polypeptides and nonrelated proteins, namely, lupin conglutin gamma and soybean Bg7S, were detected. Therefore, these proteins may be regarded as new putative allergens. The present findings show the potentiality of 2D electrophoresis in the identification of food allergens and open the way to the traceability of the new cross-reacting proteins in the food chain.


Endemic goitre prevalence in Ifedapo Local Government Area of Oyo State, Nigeria.
As part of efforts aimed at providing an adequate data base for the control of iodine deficiency disorders (IDD) in Nigeria, we undertook an extensive goitre survey of Ifedapo Local Government Area (LGA) of Oyo State, which lies within the goitre-belt of South-western Nigeria. Primary school pupils aged between six and twelve years were the subjects of the survey. From each of ten health districts of the LGA, one primary school was included in the survey. For each school, the entire school population was assessed for goitre by palpation and goitre graded according to WHO/ICCIDD criteria. The survey included 3599 pupils (1889 males and 1710 females). The total goitre rate (TGR) for the LGA was 23.4% while the visible goitre rate (VGR) was 6.4%. The highest goitre rate was recorded in District I (Sango, Saki) TGR = 39.5%; VGR = 17.3%), while the lowest rate was seen in District 4 (Wasengare) TGR = 5.2%; VGR = 0.7%). Females had higher goitre rates than males (TGR: 25.7% vs 21.3%; VGR: 7.5% vs 5.5%). Most of the goitres were Grades IA (36.7%) or IB (35.7%). Goitre grades 2 and 3 accounted for only 27.3% and 0.2%, respectively. These results show that there is a mild to moderate IDD problem in Ifedapo LGA. The implications of this finding, in view of the known deleterious consequences of IDD, are discussed and the need for correction highlighted.


Application of COVIDTracer to Inform Public Health Workforce Planning in Alaska During the COVID-19 Pandemic.
The COVID-19 pandemic has caused social and economic disruption worldwide and spurred numerous mitigation strategies, including state investments in training a large contact tracing and case investigation workforce. A team at the University of Alaska Anchorage evaluated implementation of the COVID-19 contact tracing and case investigation program of the State of Alaska Department of Health and Social Services, Division of Public Health, Section of Public Health Nursing. As part of that evaluation, the team used COVIDTracer, a spreadsheet modeling tool. COVIDTracer generated projections of COVID-19 case counts that informed estimates of workforce needs and case prioritization strategies. Case count projections approximated the reported epidemiologic curve with a median 7% difference in the first month. The accuracy of case count predictions declined after 1 month with a median difference of 80% in the second month. COVIDTracer inputs included previous case counts, the average length of time for telephone calls to cases and outreach to identified contacts, and the average number of contacts per case. As each variable increased, so too did estimated workforce needs. Decreasing the average time from exposure to outreach from 10 to 5 days reduced case counts estimated by COVIDTracer by approximately 93% during a 5-month period. COVIDTracer estimates informed Alaska's workforce planning and decisions about prioritizing case investigation during the pandemic. Lessons learned included the importance of being able to rapidly scale up and scale down workforce to adjust to a dynamic crisis and the limitations of prediction modeling (eg, that COVIDTracer was accurate for only about 1 month into the future). These findings may be useful for future pandemic preparedness planning and other public health emergency response activities.


Novel preparation of intercellular lipid models of the stratum corneum containing stereoactive ceramide.
The microstructure formed by intercellular lipids in the stratum corneum is important for the barrier function of the skin. However, the correlation between lipid composition and microstructure has not yet been clarified. To elucidate the microstructure of intercellular lipids in the stratum corneum, an intercellular lipid model was prepared from ceramide 5 (CER5), cholesterol (CHOL), and palmitic acid (PA), considering the nonuniformity of the lipid components of the stratum corneum. A response surface method incorporating thin-plate spline interpolation (RSM-S) was employed to prepare the CER5/CHOL/PA lipid bilayers. Fluorescence anisotropy of the CER5/CHOL/PA bilayers showed four distinct clusters based on Kohonen's self-organizing maps (SOM). At the centroid formulation of those clusters, the microstructures of CER5/CHOL/PA bilayers were determined using synchrotron X-ray scattering. Three kinds of lamellar structures and two kinds of lateral packing-namely, hexagonal and orthorhombic-were formed. The microstructure of the CER5/CHOL/PA bilayers was likely to be intrinsic to the intercellular lipids in the stratum corneum. In conclusion, the CER5/CHOL/PA bilayers prepared based on RSM-S and SOM were useful as models of the intercellular lipids in the stratum corneum.


Associations between variants in KITLG, SPRY4, BAK1, and DMRT1 and pediatric germ cell tumors.
Recent genome wide association studies have identified susceptibility loci for adult testicular germ cell tumors (GCT) near KITLG, SPRY4, BAK1, and DMRT1. We evaluated variants in these four genes to determine whether these are also susceptibility loci for pediatric GCTs. DNA was isolated from 52 pediatric GCTs (ages 0-21 years) obtained from the Cooperative Human Tissue Network. Control DNA was isolated from de-identified dried blood spots from 141 white newborns. Genotyping was conducted using TaqMan assays (rs4474514) or by PCR and sequencing (rs4324715, rs210138, and rs755383). Associations between variants and GCT were evaluated using logistic regression with adjustment for sex. We also evaluated whether the associations differed by age at GCT diagnosis (0-9 years, 10-21 years), sex, and tumor location (gonadal, non-gonadal). We observed a significant association for rs210138 (BAK1) and pediatric GCT overall (odds ratio (OR) = 1.80, 95% confidence interval (CI) 1.10-2.95, P = 0.02) with non-significant associations similar in magnitude in both the pediatric (P = 0.09) and adolescent (P = 0.06) age groups. The KITLG (rs4474514) and SPRY4 (rs4324715) variants were significantly associated with GCT only in the adolescent age group (rs4474514: OR = 2.28, 95% CI 1.09-4.79, P = 0.03 and rs4324715: OR = 2.40, 95% CI 1.19-4.83, P = 0.01). Associations were mostly similar when stratified by sex. This is the first study to suggest that these loci may also be important in susceptibility to GCTs in the adolescent (KITLG, SPRY4, and BAK1) and pediatric (BAK1) age groups.


Impact of native VLDL on tissue factor pathway inhibitor in endothelial cells and interactions between TFPI and lipoprotein lipase.
Tissue factor pathway inhibitor (TFPI) is a potent inhibitor of tissue factor (TF)-induced blood coagulation. A positive association between very low density lipoproteins (VLDLs) and TFPI has been reported in vivo. In contrast, one in vitro study indicates that TFPI may enhance lipoprotein lipase (LPL) activity, thereby increasing triglyceride hydrolysis. The current study was conducted to investigate how native VLDL influenced the synthesis and release of TFPI in endothelial cells, and how TFPI affected the LPL-induced hydrolysis of VLDL in vitro and at the endothelial surface. A spontaneously transformed immortal endothelial cell line (ECV304) and primary coronary artery cells (CoEc) were used, and VLDL was isolated from healthy volunteers by density gradient ultracentrifugation. Sequential free fatty acid (FFA) measurements were used to evaluate the kinetics of the LPL-induced hydrolysis. The levels of TFPI mRNAs in the stimulated cells were determined by quantitative real-time reverse transcription-ploymerase chain reaction (qPCR) using the ABI PRISM 7700 Sequence Detection System. Stimulation of ECV304 cells for 24 hours with native VLDL (0-100 microg/mL) caused a dose-dependent increase of TFPI in the medium (6.7-23.8 ng/10(6) cells, P < 0.001), without affecting the cellular content of TFPI. The expression of TFPI mRNA was significantly upregulated after 10 minutes of stimulation with n-VLDL. Both recombinant TFPI (r-TFPI) and LPL showed a dose-dependent binding to ECV 304 cells without saturation, and no competitive binding interactions between LPL and TFPI were observed at the endothelial surface. The addition of increasing concentrations of r-TFPI to ECV 304 cells, preincubated with LPL, did not affect the hydrolysis of VLDL triglycerides. The maximal reaction velocity (V(max)) of LPL-induced hydrolysis of n-VLDL was not affected by the addition of increasing concentrations of r-TFPI to the reaction mixture in vitro. The current experimental study indicates an upregulation of TFPI synthesis and release by VLDL. LPL-induced hydrolysis of VLDL in vitro was not influenced by TFPI neither in suspension nor at the endothelial surface.


Relevance of immunoglobulin G Fc receptor polymorphism to recurrence of adult periodontitis in Japanese patients.
Polymorphonuclear neutrophil (PMN) phagocytic function has been shown to be impaired in some patients with periodontitis. PMN constitutively express members of two immunoglobulin G receptor (Fc gammaR) classes: Fc gammaRIIa (CD32) and Fc gammaRIIIb (CD16). Both receptors exhibit genetically determined structural and functional biallelic polymorphisms, which have been shown to influence PMN phagocytic function. In this study, we assessed the relevance of these Fc gammaR polymorphisms to susceptibility to adult periodontitis and recurrence rate. The distribution of Fc gammaRIIa and Fc gammaRIIIb genotypes of 100 Japanese patients with adult periodontitis during follow-up was compared to the distribution of genotypes in 105 race-matched healthy controls. No significant skewing of distributions of Fc gammaRIIa and Fc gammaRIIIb genotypes was observed between patients and controls. Notably, however, a significant overrepresentation of the Fc gammaRIIIb-NA2 allotype was found in patients with disease recurrence (P < 0.05; odds ratio, 4.29; 95% confidence interval, 1.19 to 16.24). Moreover, the annual rate of recurrence was significantly higher in patients with the Fc gammaRIIIb-NA2/NA2 and Fc gammaRIIIb-NA1/NA2 genotypes than in Fc gammaRIIIb-NA1/NA1 individuals (P < 0.05). Fc gammaRIIa-R/R131 individuals also exhibited higher recurrence rates, though the difference was not statistically significant (P = 0.06). These results suggest that the Fc gammaRIIIb-NA2 allotype represents a risk factor for recurrence of adult periodontitis.


Legislating change in surgical residency.
The Patient Protection and Affordable Care Act, also known as the House of Representatives Bill HR 3590, was created to improve the quality of patient care and access to health care for American citizens. Provisions of this bill are likely to have both intended and unintended consequences on surgical education. The purpose of this article is to explore the ways in which HR 3590 may affect the educational experience of surgical house officers at teaching hospitals.


Measurement of AC magnetic field distribution using magnetic resonance imaging.
Electric currents are applied to body in numerous applications in medicine such as electrical impedance tomography, cardiac defibrillation, electrocautery, and physiotherapy. If the magnetic field within a region is measured, the currents generating these fields can be calculated using the curl operator. In this study, magnetic fields generated within a phantom by currents passing through an external wire is measured using a magnetic resonance imaging (MRI) system. A pulse sequence that is originally designed for mapping static magnetic field inhomogeneity is adapted. AC current in the form of a burst sine wave is applied synchronously with the pulse sequence. The frequency of the applied current is in the audio range with an amplitude of 175-mA rms. It is shown that each voxel value of sequential images obtained by the proposed pulse sequence is modulated similar to a single tone broadband frequency modulated (FM) waveform with the ac magnetic field strength determining the modulation index. An algorithm is developed to calculate the ac magnetic field intensity at each voxel using the frequency spectrum of the voxel signal. Experimental results show that the proposed algorithm can be used to calculate ac magnetic field distribution within a conducting sample that is placed in an MRI system.


Musculoskeletal (MSK) and Sport and Exercise Medicine (SEM) in General Practice (GP): A Novel GP-based MSK and SEM Clinic for Managing Musculoskeletal symptoms in a GP.
Musculoskeletal (MSK) complaints are common within primary care (1) (2) (3) but some General Practitioners (GPs)/family physicians do not feel comfortable managing these symptoms (3), preferring to refer onto hospital specialists or Integrated Clinical Assessment and Treatment Services (ICATs). Long waiting times for hospital outpatient reviews are a major cause of patient inconvenience and complaints (4). We therefore aimed to establish a GP-ran MSK and sport and exercise medicine (SEM) clinic based within a Belfast GP surgery that would contribute to a sustainable improvement in managing these common conditions within primary care as well as reducing waiting times for patients with these conditions to see a specialist. This shift from hospital-based to community-based management is in-keeping with recent policy changes within the UK health-system, including Transforming Your Care within Northern Ireland (NI) (5). The GP-ran MSK and SEM clinic was held monthly within a Belfast GP practice, staffed by one GP with a specialist interest in MSK and SEM conditions and its performance was reviewed over a three month period. Parameters audited included cases seen, orthopaedic and x-ray referral rates and secondary care referrals comparing the GP practice's performance to the same time period in the previous year as well as patient satisfaction questionnaires.


Visual scanning behavior during distracted walking in healthy young adults.
An epidemic of pedestrian accidents when walking while texting suggests that people are less aware of their surroundings during distracted walking, and highlights the importance of visual scanning for pedestrian safety. Quantitative examination of visual scanning during distracted walking is still lacking. Is visual scanning behavior altered by distracted walking in healthy young adults? We compared visual scanning behavior in 20 young adults during usual (single-task) walking, walking while performing a letter-fluency task, and walking while texting. Visual scanning behavior was measured by fixation count and dwell time percentage in specific areas of interest. Dual-task effects on gait speed, letter fluency, texting speed and accuracy, and situational awareness were also examined. Visual scanning behavior differed between the three walking conditions. During dual-task letter fluency, participants had significantly more non-walking path fixations than either of the other two conditions (i.e., more frequent, broader visual scanning). Conversely, during dual-task texting, gaze was focused predominantly on the phone, with little visual scanning of the walking path and surrounding environment. When walking without texting or talking, gaze was directed equally to far walking path and surrounding environment. Texting while walking is associated with a considerable reduction in overt visual attention to the walking path and surrounding areas. Whether this translates to reduced conscious awareness of environmental stimuli remains unclear. Performing a verbal task while walking was associated with more frequent, wider visual scanning behavior, which may be specific to the nature of the verbal task in this study.


Pneumonectomy: the burden of death after discharge and predictors of surgical mortality.
Pneumonectomy has the highest mortality rate among resections for lung cancer, with limited literature differentiating predictors of postpneumonectomy in-hospital mortality (IHM) from early postdischarge mortality (PDM). This study aims to examine the burden of death over time and to identify potential predictive factors, including patient comorbidities and hospital and surgeon volumes. Data were abstracted from an Ontario population-based linked database from 2005 to 2011. Proportional mortality and cumulative survival attributable to IHM and 90-day PDM is reported. Logistic and Cox regression analyses examined the role of potential factors related to death. Odds ratios (ORs) and hazard ratios (HRs) and 95% confidence intervals (CIs) were reported. Of 505 patients who underwent pneumonectomy, the median length of stay was 6 days (1-30 days). IHM was 4.4% (2.9%-6.5%), and 90-day PDM was an additional 6.4% (4.6%-9.0%). Logistic regression showed that congestive heart failure (CHF) (OR, 23.5; range, 4.0-136.0), cerebrovascular disease (OR, 12.5; range, 1.2-128.0), renal disease (OR, 8.8; range, 1.3-60.5), and previous myocardial infarction (MI) (OR, 5.4; range, 1.5-20.0) were predictive of IHM, whereas age (HR, 1.4; range, 1.1-1.7) per year and CHF (HR, 18.0; range, 4.0-79.0) were predictive of PDM. All other factors were not significant. PDM represents a distinct and underrecognized burden of postoperative death. More than half of postpneumonectomy mortality occurred after discharge, and the rate remained unchanged over the study period. Patient factors play a major role in both IHM and PDM, whereas institutional and physician volume do not influence outcome, suggesting the importance of patient selection and the need for continued evaluation of mortality.


Paralysis of ventilated newborn babies does not influence resistance of the total respiratory system.
Paralysis with pancuronium bromide is used in newborn infants to facilitate ventilatory support during respiratory failure. Changes in lung mechanics have been attributed to paralysis. The aim of this study was to examine whether or not paralysis per se has an influence on the passive respiratory mechanics, resistance (Rrs) and compliance (Crs) of the respiratory system in newborn infants. In 30 infants with acute respiratory failure, Rrs was measured during paralysis with pancuronium bromide and after stopping pancuronium bromide (group A). Rrs was also measured in an additional 10 ventilated infants in a reversed fashion (group B): Rrs was measured first in nonparalysed infants and then they were paralysed, mainly for diagnostic procedures, and the Rrs measurement repeated. As Rrs is highly dependent on lung volume, several parameters, that depend directly on lung volume were recorded: inspiratory oxygen fraction (FI,O2), arterial oxygen tension/alveolar oxygen tension (a/A) ratio and volume above functional residual capacity (FRC). In group A, the Rrs was not different during (0.236+/-0.09 cmH2O x s x mL(-1)) and after (0.237+/-0.07 cmH2O x s x mL(-1)) paralysis. Also, in group B, Rrs did not change (0.207+/-0.046 versus 0.221+/-0.046 cm x s x mL(-1) without versus with pancuronium bromide). FI,O2, a/A ratio and volume above FRC remained constant during paralysis. These data demonstrate that paralysis does not influence the resistance of the total respiratory system in ventilated term and preterm infants when measured at comparable lung volumes.


[A monitoring system for occupational cancer].
The Italian Occupation and Safety Act (d.lgs 626/94) provided for the establishment of a nationwide occupational cancer registry, under the National Institute for Occupational Health and Safety (ISPESL), with the aim of detecting cancer cases of occupational origin and estimating the influence of occupation in cancer causation. Information on cancer cases, drawn from six Italian population-based cancer registries (CRs of Friuli Venezia Giulia Region, Genoa Town and Genoa Province, Macerata Town, Umbria Region, Varese Town, Veneto Region), and on a random sample of population controls selected in each CRs area were linked with data on subjects employed in private enterprises that have been available in electronic form since 1974 at the National Institute for Social Security (INPS). In this way, both for cases and controls, the occupational histories of past employment were collected. A population-based case-control study covering the period 1990-1998 was carried out with the aim of estimating occupational cancer risk in the private sector by site and economic category in each area. Since one of the major drawbacks of this approach is the difficulty in distinguishing true occupational hazards from incidental findings derived from multiple comparisons, an extensive research of occupational literature was carried out, independently of the study results, to compare our results with existing knowledge on occupational risks. Pooled analysis of the most recent incidence data based on 36,379 cases and 29,572 controls was performed; 34 "statistically significant" associations were found for 11 economic categories. Using our literature review, 10 associations were supported by more than 5 publishedpapers, 14 by a number of papers between 1 and 5, and 10 associations had not been previously reported. This system appears suitable for assessing existing occupational cancer risks and can eventually lead to detecting occupational hazards in many areas of Italy. The system can also provide a list of cases suitable for in-depth search for past occupational exposures.


Welfare of cats 5-29 months after perineal urethrostomy: 74 cases (2015-2017).
The objective of this study was to evaluate the long-term quality of life in cats following perineal urethrostomy. This study comprised a retrospective case series of 74 cats with urinary obstructions that received perineal urethrostomies at the American Society for the Prevention of Cruelty to Animals (ASPCA) Animal Hospital between September 2015 and July 2017. Medical records were reviewed for information on the number of obstructions prior to surgery, urinary tract problems at the time of surgery and other potential factors influencing long-term welfare. Owners were surveyed by telephone and text 5-29 months after their cat's surgery. Responses were compiled and analyzed to determine owner perception of welfare postsurgery, and to identify patterns in medical history and welfare outcomes. In total, 105 cat owners were eligible for the survey; 74 responded. Altogether, 100% of responding individuals reported at least the same quality of life postsurgery compared with the cat's quality of life before demonstrating urinary problems (48% reported better). On a quality-of-life scale of 1-10 (10 = excellent), 100% of responding cat owners reported at least 7; 75% reported 10. Quality-of-life outcomes for cats 5-29 months after receiving perineal urethrostomy surgery were reported to be very good. Given this finding, and that perineal urethrostomy surgery should mitigate future blockages, we suggest considering perineal urethrostomy surgery as a standard tool to manage urinary obstructions in cats. The needs of the specific client and patient, including risks, costs and welfare of the cats, should drive the options for management of urinary obstructions in male cats.


Heat shock protein 90 is required for conidiation and cell wall integrity in Aspergillus fumigatus.
Heat shock protein 90 (Hsp90) is a eukaryotic molecular chaperone. Its involvement in the resistance of Candida albicans to azole and echinocandin antifungals is well established. However, little is known about Hsp90's function in the filamentous fungal pathogen Aspergillus fumigatus. We investigated the role of Hsp90 in A. fumigatus by genetic repression and examined its cellular localization under various stress conditions. Failure to generate a deletion strain of hsp90 suggested that it is essential. Genetic repression of Hsp90 was achieved by an inducible nitrogen-dependent promoter (pniiA-Hsp90) and led to decreased spore viability, decreased hyphal growth, and severe defects in germination and conidiation concomitant with the downregulation of the conidiation-specific transcription factors brlA, wetA, and abaA. Hsp90 repression potentiated the effect of cell wall inhibitors affecting the β-glucan structure of the cell wall (caspofungin, Congo red) and of the calcineurin inhibitor FK506, supporting a role in regulating cell wall integrity pathways. Moreover, compromising Hsp90 abolished the paradoxical effect of caspofungin. Pharmacological inhibition of Hsp90 by geldanamycin and its derivatives (17-AAG and 17-DMAG) resulted in similar effects. C-terminal green fluorescent protein (GFP) tagging of Hsp90 revealed mainly cytosolic distribution under standard growth conditions. However, treatment with caspofungin resulted in Hsp90 accumulation at the cell wall and at sites of septum formation, further highlighting its role in cell wall stress compensatory mechanisms. Targeting Hsp90 with fungal-specific inhibitors to cripple stress response compensatory pathways represents an attractive new antifungal strategy.


Increased autumn productivity permits temperate trees to compensate for spring frost damage.
Climate warming is leading to earlier budburst and therefore an increased risk of spring frost injury to young leaves. But to what extent are second-cohort leaves, which trees put out after leaf-killing frosts, able to compensate incurred losses? To investigate whether second-cohort leaves behave differently from first-cohort leaves, we exposed saplings of beech (Fagus sylvatica), oak (Quercus robur), and honeysuckle (Lonicera xylosteum) to experimental treatments mimicking either a warm spring or a warm spring with a leaf-killing frost. Refoliation took 48, 43, and 36 d for beech, oak and honeysuckle, respectively. In beech and oak, autumn Chl content and photosynthesis rates were higher in second- than in first-cohort leaves, senescence in second-cohort leaves occurred c. 2-wk-later, and autumn bud growth in beech was elevated 66% in frost-damaged plants compared with the warm spring treatment. No differences in autumn phenology and growth were observed for honeysuckle. Overall, in beech and oak, delayed Chl breakdown in second-cohort leaves mitigated 31% and 25%, respectively, of the deficit in growing-season length incurred by spring frost damage. These results reveal an unexpected ability of second-cohort leaves of beech and oak to compensate for spring frost damage, and demonstrate that long-lived trees vary their autumnal phenology depending on preceding productivity.


Predictors of shorter-, medium-, and longer-term outcome in schizophrenia.
Predictors of outcome were identified for 163 patients with DSM-III schizophrenia divided into three cohorts by length of follow-up interval: 0-9 years (N = 57), 10-19 years (N = 59), and 20 years or more (N = 47). The most powerful variables predicting outcome differed between follow-up intervals. Characteristics of premorbid functioning were most influential in the first decade of follow-up, family functioning emerged as important in the second decade, and family genetics influenced the third decade and beyond. Signs and symptoms proved predictive in consistent ways for midrange and longer-term outcomes. The results demonstrate significant variability in both the type and strength of relevant predictors depending on follow-up length.


Intramammary immunization with ultraviolet-killed Escherichia coli shows partial protection against late gestation intramammary challenge with a homologous strain.
The objective of this study was to evaluate the efficacy of intramammary immunization with UV-killed Escherichia coli ECC-Z on prevention of intramammary colonization after a challenge with a dose of the homologous E. coli ECC-Z live bacteria. A total of 10 cows were included in a study to evaluate the efficacy of intramammary immunization. All 10 cows received an intramammary immunization of 100 cfu of UV-killed E. coli ECC-Z bacteria into one hind quarter at the time of dry off. Approximately 2wk before the anticipated calving date, both hind quarters of all cows were challenged with 100 cfu of live E. coli ECC-Z bacteria. Five of the cows were vaccinated parenterally with a commercial J5 bacterin, and 5 cows served as controls with no parenteral vaccination. The cows were then followed over time and infection risk, clinical scores, somatic cell count, and milk production were observed over time. The results of these 10 cows showed partial protection of intramammary immunization on the outcome of a subsequent homologous intramammary challenge. Immunization resulted in a lower probability of infection, a lower bacteria count, lower somatic cell counts and milk conductivity, a lower clinical mastitis score, and increased milk production compared with unimmunized control quarters. Once the analysis was corrected for immunization, parenteral J5 vaccination had no significant effect on any of the measured parameters. These results provide the first evidence that intramammary immunization may improve the outcome of an intramammary E. coli infection in late gestation and onset of mastitis immediately following parturition. Unlike systemic vaccination, which generally does not reduce the intramammary infection risk, the intramammary immunization did show a 5-times reduced odds of an established intramammary infection after challenge. Cytokine profiles indicated a local return of proinflammatory response after challenge as the data showed a more pronounced increase in in IFN-γ with a subsequent negative feedback due to a spike in the level of IL-10 in immunized quarters relative to nonimmunized quarters. Although these results are preliminary and obtained on only 10 cows, the results provide insight into the biological benefits of triggering mucosal immunity in the mammary gland.


Pyrocinchonimides Conjugate to Amine Groups on Proteins via Imide Transfer.
Advances in bioconjugation, the ability to link biomolecules to each other, small molecules, surfaces, and more, can spur the development of advanced materials and therapeutics. We have discovered that pyrocinchonimide, the dimethylated analogue of maleimide, undergoes a surprising transformation with biomolecules. The reaction targets amines and involves an imide transfer, which has not been previously reported for bioconjugation purposes. Despite their similarity to maleimides, pyrocinchonimides do not react with free thiols. Though both lysine residues and the N-termini of proteins can receive the transferred imide, the reaction also exhibits a marked preference for certain amines that cannot solely be ascribed to solvent accessibility. This property is peculiar among amine-targeting reactions and can reduce combinatorial diversity when many available reactive amines are available, such as in the formation of antibody-drug conjugates. Unlike amides, the modification undergoes very slow reversion under high pH conditions. The reaction offers a thermodynamically controlled route to single or multiple modifications of proteins for a wide range of applications.


A national evaluation using standardised patient actors to assess STI services in public sector clinical sentinel surveillance facilities in South Africa.
Quality concerns in STI service delivery and missed opportunities for integration with HIV testing and prevention services in South Africa have been well documented. This national evaluation aimed to evaluate current utilisation and adherence to national STI guidelines, including partner notification and integration with HIV services, for diagnosis and management of STIs. Facility surveys assessed infrastructure and resource availability, and standardised patient (SP) assessments evaluated quality of STI care in 50 public clinics in nine provinces in South Africa. The primary outcome was the proportion of SPs receiving essential STI care, defined as: offered an HIV test, condoms, partner notification counselling and correct syndromic treatment. Weighted proportions were generated, and SP findings were compared by gender using χ2 tests with Rao-Scott correction. More than 80% of facilities reported medications in stock, with the exceptions of oral cefixime (48.3%), oral erythromycin (75.1%) and paediatric syrups. Among 195 SP encounters, 18.7% (95% CI 10.7% to 30.5%) received all hypothesised essential STI services: offered HIV test (67.1%), offered condoms (31.4%), partner notification counselling (70.2%) and recommended syndromic treatment (60.7%). Men were more likely than women to be offered all services (25.1% vs 12.3%, p=0.023), recommended treatment (70.7% vs 50.9%, p=0.013) and partner notification counselling (79.9% vs 60.6%, p=0.020). Only 6.3% of providers discussed male circumcision with male SPs, and 26.3% discussed family planning with female SPs. This evaluation of STI services across South Africa found gaps in the availability of medications, adherence to STI guidelines, condom provision and prevention messaging. Limited integration with HIV services for this high-risk population was a missed opportunity. Quality of STI care should continue to be monitored, and interventions to improve quality should be prioritised as part of national strategic HIV and primary healthcare agendas.


Negative Regulation of mTOR Signaling Restricts Cell Proliferation in the Floor Plate.
The neural tube is composed of a number of neural progenitors and postmitotic neurons distributed in a quantitatively and spatially precise manner. The floor plate, located in the ventral-most region of the neural tube, has a lot of unique characteristics, including a low cell proliferation rate. The mechanisms by which this region-specific proliferation rate is regulated remain elusive. Here we show that the activity of the mTOR signaling pathway, which regulates the proliferation of the neural progenitor cells, is significantly lower in the floor plate than in other domains of the embryonic neural tube. We identified the forkhead-type transcription factor FoxA2 as a negative regulator of mTOR signaling in the floor plate, and showed that FoxA2 transcriptionally induces the expression of the E3 ubiquitin ligase RNF152, which together with its substrate RagA, regulates cell proliferation via the mTOR pathway. Silencing of RNF152 led to the aberrant upregulation of the mTOR signal and aberrant cell division in the floor plate. Taken together, the present findings suggest that floor plate cell number is controlled by the negative regulation of mTOR signaling through the activity of FoxA2 and its downstream effector RNF152.


Reduced plasma concentrations of vitamin B6 and increased plasma concentrations of the neurotoxin 3-hydroxykynurenine are associated with nodding syndrome: a case control study in Gulu and Amuru districts, Northern Uganda.
Nodding syndrome was first reported in Uganda in 2003 among internally displaced populations. Risk factors for the syndrome remain unknown. We therefore explored vitamin B6 deficiency and resulting high 3-hydroxykynurenine (3-HK) levels as risk factor for nodding syndrome in Northern Uganda. Case-control study conducted in Gulu and Amuru districts. Cases were children/young adults with nodding syndrome. Healthy children/young adults were recruited as controls from same community as cases. Data on socio-demographic and other risk factors was collected using questionnaires. Whole blood was collected in EDTA tubes for assay of 3-HK and vitamin B6 using sandwich ELISA. Conditional logistic regression model was used to assess associations. 66 cases and 73 controls were studied. Factors associated with nodding syndrome were being positive for 3-HK (AOR=4.50, p=0.013), vitamin B6 concentration below mean (AOR=7.22, P=0.001), child being taken care of by mother only (AOR=5.43, p=0.011), child being taken care of by guardian (AOR=5.90, p=0.019) and child consuming relief food at weaning (AOR=4.05, p=0.021). Having low vitamin B6 concentration which leads to a build up of 3-hydroxykynurenine concentration in cases as a main risk factor. Therefore, cases should be treated with vitamin B6 and community members should be sensitise to ensure adequate dietary intake of vitamin B6 so that the risk of nodding syndrome among children is averted. We encourage future prospective intervention study to be conducted to assess the effect of low vitamin B6 on the development of nodding syndrome via raised 3-HK concentration.


Chemical probes of Skp2-mediated p27 ubiquitylation and degradation.
Skp2 is a member of the F-box family of proteins that serve as substrate-specific adaptors in Skp1-CUL1-ROC1-F-box (SCF) E3 ubiquitin ligases. Skp2 (Fbxl1) directly binds to the tumor suppressor p27 in the context of the SCFSkp2 E3 ubiquitin ligase to ubiquitylate and target-phosphorylated p27 for proteasomal degradation. As p27 is a powerful suppressor of growth in a variety of cells, and as Skp2 is also overexpressed in many human cancers, Skp2 is considered an oncogene and an intriguing drug target. However, despite 20 years of investigation, a valid chemical inhibitor of Skp2-mediated degradation of p27 has not been identified. Recently, an increasing number of compounds designed to have this bioactivity have been reported. Here, we conduct a meta-analysis of the evidence regarding bioactivity, structure, and medicinal chemistry in order to evaluate and compare these Skp2 inhibitor compounds. Despite chemically diverse compounds with a wide array of Skp2-mediated p27 ubiquitylation inhibition properties reported by several independent groups, no current chemical probe formally qualifies as a validated pharmaceutical hit compound. This finding suggests that our knowledge of the structural biochemistry of the Skp2-p27 complex remains incomplete and highlights the need for novel modes of inquiry.


Isolation and structure of nocobactin NA, a lipid-soluble iron-binding compound from Nocardia asteroides.
Nocobactin NA, a lipid-soluble iron-chelating product with an unusual and characteristic u.v.-absorption spectrum, was isolated from Nocardia asteroides grown under conditions of iron deficiency. Its structure was determined by physical methods and by synthesis of one of its degradation products. Nocobactin NA was obtained as a homologous mixture of compounds with side chains of differing length, and resembles mycobactin M in structure except that it has an oxazole ring in place of an oxazoline ring, and the side chains in the cobactin fragment are considerably shorter.


Chromatin structure in telomere dynamics.
The establishment of a specific nucleoprotein structure, the telomere, is required to ensure the protection of chromosome ends from being recognized as DNA damage sites. Telomere shortening below a critical length triggers a DNA damage response that leads to replicative senescence. In normal human somatic cells, characterized by telomere shortening with each cell division, telomere uncapping is a regulated process associated with cell turnover. Nevertheless, telomere dysfunction has also been associated with genomic instability, cell transformation, and cancer. Despite the essential role telomeres play in chromosome protection and in tumorigenesis, our knowledge of the chromatin structure involved in telomere maintenance is still limited. Here we review the recent findings on chromatin modifications associated with the dynamic changes of telomeres from protected to deprotected state and their role in telomere functions.


Pain at the vulvar vestibule: a web-based survey.
To assess the prevalence, characteristics, and ethnic distribution of pain in the vulvar vestibule among a national sample of women completing a web-based survey. Female participants of the SurveySpot Internet research panel (Survey Sampling International) were invited via e-mail to respond to a web-based questionnaire on women's health issues. The frequency and characteristics of reported pain of the vulvar vestibule were assessed among 994 respondents, using univariate and multivariate analyses. Between May 24, 2002, and June 6, 2002, 730 non-African American women and 364 African American women responded to the invitation to participate, and 94.5% completed the survey. A history of pain at the vulvar vestibule was reported by 288 women (27.9%), with 80 (7.8% of the initial 1032) reporting pain within the past 6 months, 31 (3.0%) reporting pain that lasted 3 or more months, and 18 (1.7%) reporting vestibular pain lasting 3 or more months that occurred within the past 6 months. The prevalence of pain was similar in African American and non-African American women. The prevalence of pain at the vulvar vestibule is more common than previously estimated. The perception that vulvar pain is rare among African American women was not supported in this survey.


Primary malignant lymphoma of the cervix uteri: a case report.
Primary reticuloendothelial disease of the genital tract is an extremely rare condition. We encountered one such patient with diffuse large cell malignant lymphoma of the cervix uteri who presented with irregular vaginal bleeding. After complete haematological and radiological investigations, satisfactory treatment of the Stage I disease was achieved with total hysterectomy, bilateral salpingo-oophorectomy and post-operative pelvic irradiation.


Insights into colloid-mediated trace element release at the soil/water interface.
Organic or inorganic colloids play a major role in the mobilization of trace elements in soils and waters. Environmental physicochemical parameters (pH, redox potential, temperature, pressure, ionic strength, etc.) are the controlling factors of the colloidal mobilization. This study was dedicated to follow the colloid-mediated mobilization of trace elements through time at the soil/water interface by means of an experimental approach. Soil column experiments were carried out using percolating synthetic solutions. The percolated solutions were ultrafiltrated with various decreasing cutoff thresholds to separate the different colloidal phases in which the dissolved organic carbon and trace element concentrations were measured. The major results which stem from this study are the following: (i) The data can be divided into different groups of organic compounds (microbial metabolites, fulvic acids, humic acids) with regard to their respective aromaticity and molecular weight. (ii) Three groups of elements can be distinguished based on their relationships with the colloidal phases: the first one corresponds to the so-called "truly" dissolved group (Li, B, K, Na, Rb, Si, Mg, Sr, Ca, Mn, Ba, and V). The second one can be considered as an intermediate group (Cu, Cd, Co, and Ni), while the third group gathers Al, Cr, U, Mo, Pb, Ti, Th, Fe, and rare earth elements (REE) carried by the organic colloidal pool. (iii) The data demonstrate that the fulvic acids seem to be a major organic carrier phase for trace elements such as Cu, Cd, Co, and Ni. By contrast, the trace elements belonging to the so-called colloidal pool were mostly mobilized by humic acids containing iron nanoparticles. Lead, Ti, and U were mobilized by iron nanoparticles bound to these humic acids. Thus, humic substances allowed directly or indirectly a colloidal transport of many insoluble trace elements either by binding trace elements or by stabilizing a ferric carrier phase. (iv) Finally, the results demonstrated also that REE were mostly mobilized by humic substances. The REE normalized patterns showed a middle REE downward concavity. Therefore, as previously shown elsewhere humic substances are a major control of REE speciation and REE fractionation patterns as well since the humic substance/metal ratio was the key parameter controlling the REE pattern shape.


Corticosteroids increase intracellular free sodium ion concentration via glucocorticoid receptor pathway in cultured neonatal rat cardiomyocytes.
Glucocorticoids as well as mineralocorticoid have been shown to play essential roles in the regulation of electrical and mechanical activities in cardiomyocytes. Excess of these hormones is an independent risk factor for cardiovascular disease. Intracellular sodium ([Na+]i) kinetics are involved in cardiac diseases, including ischemia, heart failure and hypertrophy. However, intrinsic mediators that regulate [Na+]i in cardiomyocytes have not been widely discussed. Moreover, the quantitative estimation of altered [Na+]i in cultured cardiomyocytes and the association between the level of [Na+]i and the severity of pathological conditions, such as hypertrophy, have not been precisely reported. We herein demonstrate the quantitative estimation of [Na+]i in cultured neonatal rat cardiomyocytes following 24 h of treatment with corticosterone, aldosterone and dexamethasone. The physiological concentration of glucocorticoids increased [Na+]i up to approximately 2.5 mM (an almost 1.5-fold increase compared to the control) in a dose-dependent manner; this effect was blocked by a glucocorticoid receptor (GR) antagonist but not a mineralocorticoid receptor antagonist. Furthermore, glucocorticoids induced cardiac hypertrophy, and the hypertrophic gene expression was positively and significantly correlated with the level of [Na+]i. Dexamethasone induced the upregulation of Na+/Ca2 + exchanger 1 at the mRNA and protein levels. The physiological concentration of glucocorticoids increases [Na+]i via GR. The dexamethasone-induced upregulation of NCX1 is partly involved in the glucocorticoid-induced alteration of [Na+]i in cardiomyocytes. These results provide new insight into the mechanisms by which glucocorticoid excess within a physiological concentration contributes to the development of cardiac pathology.


Resistance to fatal central nervous system disease by mouse hepatitis virus, strain JHM. II. Adherent cell-mediated protection.
Resistance of SJL/J mice to intracranial inoculation with the JHM strain of mouse hepatitis, a coronavirus, is dependent upon the age of the animals at inoculation. Animals 12 weeks of age or older are resistant, whereas those 6 weeks or younger are uniformly susceptible to viral infection. Spleen cells or thioglycolate elicited peritoneal exudate cells can transfer resistance from 12-week-old to 6-week-old recipients. Removal of the adherent cells from either spleen or peritoneal cells ablated protection. Adherent cells from 12-week-old mice were protective even after depletion of Ia- and Thy-1-bearing cells. Antiviral antibody, thioglycolate injection into 6-week-old animals, and nylon wool-purified T cells were ineffective in mediating resistance. Adherent cells transferred 4 days before virus challenge, but not after challenge, were protective. Thus, there is an age-related change in SJL mice that protects from acute central nervous system disease, which may be due to maturation of a specialized adherent cell population.


Not a Simple Tether: Binding of Toxoplasma gondii AMA1 to RON2 during Invasion Protects AMA1 from Rhomboid-Mediated Cleavage and Leads to Dephosphorylation of Its Cytosolic Tail.
Apical membrane antigen 1 (AMA1) is a receptor protein on the surface of Toxoplasma gondii that plays a critical role in host cell invasion. The ligand to which T gondii AMA1 (TgAMA1) binds, TgRON2, is secreted into the host cell membrane by the parasite during the early stages of invasion. The TgAMA1-TgRON2 complex forms the core of the "moving junction," a ring-shaped zone of tight contact between the parasite and host cell membranes, through which the parasite pushes itself during invasion. Paradoxically, the parasite also expresses rhomboid proteases that constitutively cleave the TgAMA1 transmembrane domain. How can TgAMA1 function effectively in host cell binding if its extracellular domain is constantly shed from the parasite surface? We show here that when TgAMA1 binds the domain 3 (D3) peptide of TgRON2, its susceptibility to cleavage by rhomboid protease(s) is greatly reduced. This likely serves to maintain parasite-host cell binding at the moving junction, a hypothesis supported by data showing that parasites expressing a hypercleavable version of TgAMA1 invade less efficiently than wild-type parasites do. Treatment of parasites with the D3 peptide was also found to reduce phosphorylation of S527 on the cytoplasmic tail of TgAMA1, and parasites expressing a phosphomimetic S527D allele of TgAMA1 showed an invasion defect. Taken together, these data suggest that TgAMA1-TgRON2 interaction at the moving junction protects TgAMA1 molecules that are actively engaged in host cell penetration from rhomboid-mediated cleavage and generates an outside-in signal that leads to dephosphorylation of the TgAMA1 cytosolic tail. Both of these effects are required for maximally efficient host cell invasion. Nearly one-third of the world's population is infected with the protozoan parasite Toxoplasma gondii, which causes life-threatening disease in neonates and immunocompromised individuals. T. gondii is a member of the phylum Apicomplexa, which includes many other parasites of veterinary and medical importance, such as those that cause coccidiosis, babesiosis, and malaria. Apicomplexan parasites grow within their hosts through repeated cycles of host cell invasion, parasite replication, and host cell lysis. Parasites that cannot invade host cells cannot survive or cause disease. AMA1 is a highly conserved protein on the surface of apicomplexan parasites that is known to be important for invasion, and the work presented here reveals new and unexpected insights into AMA1 function. A more complete understanding of the role of AMA1 in invasion may ultimately contribute to the development of new chemotherapeutics designed to disrupt AMA1 function and invasion-related signaling in this important group of human pathogens.


Phosphatidylinositol-3,4,5-trisphosphate: tool of choice for class I PI 3-kinases.
Class I PI 3-kinases signal by producing the signaling lipid phosphatidylinositol(3,4,5) trisphosphate, which in turn acts by recruiting downstream effectors that contain specific lipid-binding domains. The class I PI 3-kinases comprise four distinct catalytic subunits linked to one of seven different regulatory subunits. All the class I PI 3-kinases produce the same signaling lipid, PIP3, and the different isoforms have overlapping expression patterns and are coupled to overlapping sets of upstream activators. Nonetheless, studies in cultured cells and in animals have demonstrated that the different isoforms are coupled to distinct ranges of downstream responses. This review focuses on the mechanisms by which the production of a common product, PIP3, can produce isoform-specific signaling by PI 3-kinases.


Gastrointestinal motor dysfunction, symptoms, and neuropathy in noninsulin-dependent (type 2) diabetes mellitus.
Although relatively frequent. diabetic involvement of digestive tract motility has not been investigated extensively in different organs. The authors studied esophageal, gastric, and gallbladder motor function in 35 type 2 (noninsulin-dependent) diabetic patients to determine the extent of gut involvement. Of these patients, 27 (77%) had peripheral neuropathy, 12 (34%) had both peripheral and autonomic neuropathy, and 22 (63%) had gastrointestinal symptoms. Esophageal manometric abnormalities were recorded in 18 patients, and delayed radionuclide emptying of the esophagus was documented in 16 patients, with a 83% concordance between the two tests. Scintigraphic gastric emptying of solids was delayed in 56% of patients, whereas gallbladder emptying after cholecystokinin stimulation was reduced in 69% of them. In 74% of patients at least one of the viscera under investigation showed abnormal motor function; however, only 36% of patients displayed involvement of the three organs. Gastrointestinal symptoms, duration and therapy of diabetes, previous poor glycemic control, and retinopathy did not correlate with the presence or the extent of motor disorders. Neuropathy was not predictive of gastrointestinal involvement and its extent; however, when motor abnormalities were present in patients with neuropathy, these were usually more severe. Gastrointestinal motor disorders are frequent and widespread in type 2 diabetics, regardless of symptoms. Autonomic neuropathy has a poor predictive value on motor disorders (0.75 for the esophagus, 0.5 for the stomach, 0.8 for the gallbladder), thus suggesting the coexistence of other pathophysiologic mechanisms.


Radial distribution of the refractive index in light-focusing rods: determination using Interphako interference microscopy.
An interferometric technique for the determination of the radial distribution of the refractive index in lightfocusing plastic rods (LFR) has been developed using the shearing method of Interphako interference microscopy. We have derived mathematical expressions for the shape of interference fringes across the rod assuming the index distribution of LFR is described by n(r) n(=) (1 - ar(2) + br(4) + cr(6) + dr(2)) in which a, b, c, and d are distribution constants. The dsitribution constants were determined using a least-squares technique on a computer from the actual shape of fringes. Results for the representative cases are shown.


Isolation, characterization, and functional assessment of oxidatively modified subfractions of circulating low-density lipoproteins.
Current evidence suggests that oxidatively modified human plasma low-density lipoproteins (ox-LDLs) are proatherogenic and cytotoxic to endothelial and vascular smooth muscle cells. The present study describes a method using ion-exchange chromatography that is capable of large-scale subfractionation of LDL for adequate analyses of composition or bioactivities. LDLs from normolipidemic (N-LDL) and homozygous familial hypercholesterolemic (FH-LDL) subjects were separated into 5 subfractions (L1 through L5) by high-capacity ion-exchange chromatography. The most strongly retained fraction from FH subjects, FH-L5, suppressed DNA synthesis in cultured bovine aortic endothelial cells and stimulated mononuclear cell adhesion to cultured endothelial cells under flow conditions in vitro. L5, which represented 1.1+/-0.2% and 3.7+/-1.7% of the LDL from N-LDL and FH-LDL, respectively, was more triglyceride-rich (17% versus 5%) and cholesteryl ester-poor (23% versus 33%) than were L1 through L4. Electrophoretic mobilities on agarose gels increased from L1 to L5. According to SDS-PAGE, apolipoprotein B-100 in N-LDL fractions L1 through L5 appeared as a single approximately 500-kDa band. In contrast, the fractions isolated from FH-LDL showed substantial fragmentation of the apolipoprotein B-100, including bands between 200 and 116 kDa. Competitive ELISA analyses using a malondialdehyde-specific monoclonal antibody against Cu2+ ox-LDL suggest that FH-L5 is malondialdehyde-modified. Relative to N-LDL, FH-LDL contains higher concentrations of a fraction, L5, that exhibits distinctive physicochemical properties and biological activities that may contribute to initiation and progression of atherogenesis in vivo.


Resistin, a fat-derived secretory factor, promotes metastasis of MDA-MB-231 human breast cancer cells through ERM activation.
Resistin, an adipocyte-secreted factor, is known to be elevated in breast cancer patients. However, the molecular mechanism by which resistin acts is not fully understood. The aim of this study was to investigate whether resistin could stimulate invasion and migration of breast cancer cells. Here, we report that resistin stimulated invasion and migration of breast cancer cells as well as phosphorylation of c-Src. Inhibition of c-Src blocked resistin-induced breast cancer cell invasion. Resistin increased intracellular calcium concentration, and chelation of intracellular calcium blocked resistin-mediated activation of Src. Resistin also induced phosphorylation of protein phosphatase 2A (PP2A). Inhibition of c-Src blocked resistin-mediated PP2A phosphorylation. In addition, resistin increased phosphorylation of PKCα. Inhibition of PP2A enhanced resistin-induced PKCα phosphorylation, demonstrating that PP2A activity is critical for PKCα phosphorylation. Resistin also increased phosphorylation of ezrin, radixin, and moesin (ERM). Additionally, ezrin interacted with PKCα, and resistin promoted co-localization of ezrin and PKCα. Either inhibition of c-Src and PKCα or knock-down of ezrin blocked resistin-induced breast cancer cells invasion. Moreover, resistin increased expression of vimentin, a key molecule for cancer cell invasion. Knock-down of ezrin abrogated resistin-induced vimentin expression. These results suggest that resistin play as a critical regulator of breast cancer metastasis.


Effect of Polymerization Accelerator on Bond Strength to Eugenol-Contaminated Dentin.
To evaluate the effect of a polymerization accelerator on the microtensile bond strength (µTBS) of etch-and-rinse and self-etch adhesives to eugenol-contaminated dentin. Sixty flat dentin surfaces were prepared from human molars. Half of the specimens were restored with zinc oxide eugenol temporary cement (IRM) (eugenol-contaminated group) and the other half remained without restoration (control group). After 24-h storage, the cement was mechanically removed. Then the specimens in each group were further divided into three subgroups based on the application procedure of a polymerization accelerator (p-toluenesulfinic acid sodium salt; Accel): no application, 10-s application, or 30-s application. After air drying, the dentin surfaces were bonded with either a three-step etch-and-rinse adhesive (OptiBond FL) or a two-step self-etch adhesive (Clearfil SE Bond) and restored with composite. After 24-h water storage, the bonded specimens were subjected to the µTBS test. Data were analyzed by three-way ANOVA and Dunnett's T3 test (p < 0.05). The eugenol-contaminated groups had significantly lower µTBS than the control groups with both types of adhesives (p < 0.05), and the application of Accel significantly increased the compromised µTBS to eugenol-contaminated dentin. Optibond FL presented significantly higher µTBS to eugenol-contaminated dentin than did Clearfil SE Bond (p < 0.05). The application of a polymerization accelerator on eugenol-contaminated dentin prior to adhesive resin application increased the μTBS of both the three-step etch-and-rinse and two-step self-etch adhesive.


Studies on Proteins of simian sarcoma-associated virus with different growth history.
Simian sarcoma-associated virus (SSaV) was repeatedly passaged on three human cell lines. The proteins of the progeny virus were analysed for the presence of variant polypeptides. Occasionally, a few variant polypeptides were observed. One-dimensional peptide maps of the major virus protein p30 revealed no modifications after 25 cycles of infection on the three cell lines studied. The peptide map of Pr65gag of virus grown through 25 passages on a human chondrosarcoma cell line was slightly different from that of the virus stock before passaging. The relative amount of the virus protein p30 as compared to p18 and p16 (possibly the SSaV equivalents of p15E and p12E) was variable depending on the host cell. Virus grown on Daudi cells was relatively deficient in p18 and p16. These virus particles were morphologically altered and had a low infectivity.


Telomere length is associated with oppositional defiant behavior and maternal clinical depression in Latino preschool children.
Exposure to psychological stress and depression are associated with shorter white blood cell telomere length (TL) in adults, possibly via associated lifelong oxidative stressors. Exposure to maternal depression increases risk for future depression and behavior problems in children, and Latino youth are at high risk. Few studies have evaluated the role of exposure to maternal depression or child behavior in relation to TL in children. We assessed early-childhood exposures to maternal depression from birth to the age of 5 years and child behavior from ages 3-5 years in a cohort of Latino children in relation to child leukocyte TL at ages 4 and 5 years. Children who had oppositional defiant behavior at 3, 4 or 5 years had shorter TL than those without by ~450 base pairs (P < 0.01). In multivariate analyses, independent predictors for shorter TL at 4 and 5 years of age included oppositional defiant disorder at 3, 4 or 5 years (β = -359.25, 95% CI -633.84 to 84.66; P = 0.01), exposure to maternal clinical depression at 3 years of age (β = -363.99, 95% CI -651.24 to 764.74; P = 0.01), shorter maternal TL (β = 502.92, 95% CI 189.21-816.63) and younger paternal age at the child's birth (β = 24.63, 95% CI 1.14-48.12). Thus, exposure to maternal clinical depression (versus depressive symptoms) in early childhood was associated with deleterious consequences on child cellular health as indicated by shorter TL at 4 and 5 years of age. Similarly, children with oppositional defiant behavior also had shorter TL, possibly related to early exposures to maternal clinical depression. Our study is the first to link maternal clinical depression and oppositional defiant behavior with shorter TL in the preschool years in a relatively homogenous population of low-income Latino children.


[Curricular change: a challenge for an undergraduate nursing course].
This work presents a research that intended to contribute with the evaluation of the Pedagogic Political Project (PPP) of an undergraduate nursing course. The objective is to analyze and to argue the curricular reformularization; contributing to the process of discussion and evaluation. The used methodology was the comparative analysis between the PPP of 1995 and 2003 and the data collected with professors and students concerning the theoretical presuppositions of the course that it was the identified innovative element in the PPP of 2003. The data revealed that the theoretical presuppositions, inyour majority guided the formation process, but there are concepts that still little incorporate like the interdisciplinarity. Another raised data was the non-effective participation of students and professors in the process of implantation and evaluation of the PPP.