Cryopreservation of rat islets of Langerhans: a comparison of two techniques.
Although a number of different protocols have been described for the cryopreservation of pancreatic islets of Langerhans, most involve equilibration with 2 M Me2SO followed by slow cooling and fast warming. This paper compares two methods with identical equilibration protocols: one with cooling at 0.25 degree C/min to -40 degrees C prior to transferring to liquid nitrogen and subsequent warming at 200 degrees C/min and sucrose dilution of Me2SO (Method A) and the other with cooling at 0.3 degree C/min to -70 degrees C prior to transferring to liquid nitrogen and warming at 50 degrees C/min followed by stepwise dilution of Me2SO (Method B). Islet viability was assessed by syngeneic transplantation to the renal subcapsular space of streptozotocin-induced diabetic rats. Seven hundred fifty fresh islets in the size range 100-200 microns consistently reversed diabetes (blood glucose < 10 mmol/liter) whereas 1000 Method A cryopreserved islets and 2000 Method B cryopreserved islets were required. Cryopreservation by either method resulted in impaired islet function compared to that of fresh islets, although this functional loss could be partially compensated by the transplantation of greater numbers of cryopreserved islets.


Radiographic changes of the proximal third metatarsal bone do not predict presence or severity of proximal suspensory desmopathy in a predominately Quarter Horse population.
Comparison of radiography to magnetic resonance imaging (MRI) can help objectively assess the value and limitations of radiographs in orthopaedic disease processes. The tarsus and proximal suspensory origin of the metatarsus are commonly imaged regions for lameness. The knowledge gained by comparison between imaging modalities will aid in improving accuracy of radiographic interpretation. To compare the radiographic and MRI findings of the proximal third metatarsal bone (MTIII) and proximal suspensory ligament using MRI as the gold standard. Retrospective observational study. Single hindlimbs of 35 horses with radiographic and high-field (3Tesla (3T)) MRI studies were blindly evaluated by two board-certified veterinary radiologists and a radiology resident. Severity and location of the following parameters were assessed: radiographic MTIII sclerosis and lucent regions, MRI endosteal metatarsal sclerosis and bone marrow lesions, plantar cortical proliferation and resorption, and proximal suspensory desmopathy (PSD). Radiographic osseous changes of MTIII were identified in 54% of limbs, whereas 40% limbs had osseous changes on MRI: 43% of limbs had PSD on MRI. No significant association was found between the presence of radiographic changes in MTIII and PSD on MRI (P = .7). A statistically significant association and positive correlation was found between the severity of radiographic changes and MTIII plantar cortical proliferation and resorption on MRI (P = .01). Retrospective study with no histopathology. Ability to correlate findings with lameness was limited by the lack of standardised time intervals between onset of lameness, diagnostic analgesia and advanced imaging. Radiographic bony changes of proximal MTIII do not reliably predict presence or severity of PSD but are associated with osseous changes at the proximal MTIII. The presence of radiographic changes should warrant additional diagnostics in the region prior to making conclusions about the presence or absence of PSD.


Contacts With the Health Care System Before Out-of-Hospital Cardiac Arrest.
Background It remains challenging to identify patients at risk of out-of-hospital cardiac arrest (OHCA). We aimed to examine health care contacts in patients before OHCA compared with the general population that did not experience an OHCA. Methods and Results Patients with OHCA with a presumed cardiac cause were identified from the Danish Cardiac Arrest Registry (2001-2014) and their health care contacts (general practitioner [GP]/hospital) were examined up to 1 year before OHCA. In a case-control study (1:9), OHCA contacts were compared with an age- and sex-matched background population. Separately, patients with OHCA were examined by the contact type (GP/hospital/both/no contact) within 2 weeks before OHCA. We included 28 955 patients with OHCA. The weekly percentages of patient contacts with GP the year before OHCA were constant (25%) until 1 week before OHCA when they markedly increased (42%). Weekly percentages of patient contacts with hospitals the year before OHCA gradually increased during the last 6 months (3.5%-6.6%), peaking at the second week (6.8%) before OHCA; mostly attributable to cardiovascular diseases (21%). In comparison, there were fewer weekly contacts among controls with 13% for GP and 2% for hospital contacts (P<0.001). Within 2 weeks before OHCA, 57.8% of patients with OHCA had a health care contact, and these patients had more contacts with GP (odds ratio [OR], 3.17; 95% CI, 3.09-3.26) and hospital (OR, 2.32; 95% CI, 2.21-2.43) compared with controls. Conclusions The health care contacts of patients with OHCA nearly doubled leading up to the OHCA event, with more than half of patients having health care contacts within 2 weeks before arrest. This could have implications for future preventive strategies.


Characterization of key aroma compounds from different rose essential oils using gas chromatography-mass spectrometry, gas chromatography-olfactometry and partial least squares regression.
To characterise the key aroma compounds of rose essential oils, five samples (Rosa damascena essential oil, Rosa centifolia essential oil, Rosa alba essential oil, Rosa rugosa cv. 'Plena' essential oil, Rosa xanthina Lindl essential oil) were analysed by gas chromatography-olfactometry (GC-O), gas chromatography-mass spectrometry (GC-MS) and quantitative descriptive analysis (QDA). Thirty-nine aroma compounds were selected as key aroma compounds by GC-MS and GC-O. The aroma of rose essential oils was described by 10 sensory attributes such as honey, sweet, fermented, spicy, fruity, woody, floral, herbal, green and fresh. The partial least squares regression (PLSR) result showed the relationship between key aroma compound and characteristic aromas of rose essential oils. This paper provided a reference for the flavourists.


Robustness of plant-insect herbivore interaction networks to climate change in a fragmented temperate forest landscape.
Forest fragmentation and climate change are among the most severe and pervasive forms of human impact. Yet, their combined effects on plant-insect herbivore interaction networks, essential components of forest ecosystems with respect to biodiversity and functioning, are still poorly investigated, particularly in temperate forests. We addressed this issue by analysing plant-insect herbivore networks (PIHNs) from understories of three managed beech forest habitats: small forest fragments (2.2-145 ha), forest edges and forest interior areas within three continuous control forests (1050-5600 ha) in an old hyper-fragmented forest landscape in SW Germany. We assessed the impact of forest fragmentation, particularly edge effects, on PIHNs and the resulting differences in robustness against climate change by habitat-wise comparison of network topology and biologically realistic extinction cascades of networks following scores of vulnerability to climate change for the food plant species involved. Both the topological network metrics (complexity, nestedness, trophic niche redundancy) and robustness to climate change strongly increased in forest edges and fragments as opposed to the managed forest interior. The nature of the changes indicates that human impacts modify network structure mainly via host plant availability to insect herbivores. Improved robustness of PIHNs in forest edges/small fragments to climate-driven extinction cascades was attributable to an overall higher thermotolerance across plant communities, along with positive effects of network structure. The impoverishment of PIHNs in managed forest interiors and the suggested loss of insect diversity from climate-induced co-extinction highlight the need for further research efforts focusing on adequate silvicultural and conservation approaches.


[Erdheim-Chester disease. Apropos of a case with autopsy findings].
Erdheim-Chester's Disease is a very uncommon variety of non-Langerhans histiocytosis of unknown etiology, which characteristically affects long bones bilaterally and symmetrically in adults. It may be accompanied by visceral foci of variable localization and extension determining prognosis. Bone scintigraphy is characteristic enough to evoke the disease but histologic examination of a peripheral specimen is required to confirm the diagnosis: spumous histiocytes CD68+, PS100+/-, CD1a-. We describe a case revealed by a severe lung disease with detailed autopsy.


Gunshot wounds: epidemiology, wound ballistics, and soft-tissue treatment.
The extremities are the most common anatomic location for gunshot wounds. Because of the prevalence of gunshot injuries, it is important that orthopaedic surgeons are knowledgeable about caring for them. The most common injuries seen with gunshot wounds are those of the soft tissues. Nonsurgical management of patients who have gunshot wounds with minimal soft-tissue disruption has been successfully accomplished in emergency departments for several years; this includes extremity wounds without nerve, intra-articular, or vascular injury. Stable, nonarticular fractures of an extremity have also been successfully treated with either minimal surgical or nonsurgical methods in the emergency department. Indications for surgical treatment include unstable fractures, intra-articular injuries, a significant soft-tissue injury (especially with skin loss), vascular injury, and/or a large or expanding hematoma.


A comprehensive acute coronary syndrome algorithm for centers with percutaneous coronary intervention capability.
A comprehensive acute coronary syndrome (ACS) protocol was developed to improve the quality of care for patients admitted with definite or probable ACS. These protocols were constructed to streamline the practice for diverse clinicians who care for ACS patients across a variety of clinical settings. They are applicable in the emergency department, the cardiac catheterization laboratory, and the inpatient settings for hospitals with primary percutaneous coronary intervention capability. These protocols standardized the care by selecting the best therapy for each clinical scenario based on available established guidelines to insure the safest and highest value (quality/cost) medical care.


Interorgan ammonia metabolism in health and disease: a surgeon's view.
Ammonia is a toxic molecule that is the principal by-product of amino acid metabolism. Although the transport of ammonia in a nontoxic form protects the brain against high circulating levels, the interorgan transport of this molecule and the orchestration between tissues that has evolved is related primarily to the fact that the nitrogen molecule is an essential molecule for the maintenance of the body's nutrition economy and overall metabolic homeostasis. Efficient handling and disposal of ammonia requires a cooperative effort between tissues in order to maintain nitrogen homeostasis. The liver is the central organ of ammonia metabolism, but other organs also play a key role in the interorgan exchange of this molecule. Alterations in ammonia metabolism occur during critical illness. These changes are adaptive and are designed to maintain metabolic homeostasis. Interorgan cooperation in ammonia metabolism is necessary to insure the proper integration of the metabolic processes which contribute to and are essential for survival during critical illness. An understanding of these processes improves our knowledge of metabolic regulation and will lead to a rational approach to the nutritional and metabolic support provided to critically ill patients.


The pathogenesis of HIV-associated nephropathy.
HIV-associated nephropathy (HIVAN) is one of the leading causes of ESRD in HIV-1-seropositive patients. Patients typically present with heavy proteinuria and chronic renal failure with pathologic findings of collapsing focal segmental glomerulosclerosis (FSGS). The disease is caused by direct infection of renal epithelial cells by HIV-1 in a genetically susceptible host. The genetic factors responsible for the susceptibility to HIVAN among blacks include a noncoding variant in the podocyte-expressed gene nonmuscle myosin, heavy chain 9 (MYH9) as well as other genes yet to be identified. Podocyte and tubular dysfunction results from the expression of viral genes, in particular nef and vpr, and the subsequent dysregulation of numerous host factors, including critical signaling pathways, inflammatory mediators, and others. The identification of these factors has the potential to provide novel therapeutic targets to prevent and treat this important disease.


Risky assessments: participant suicidality and distress associated with research assessments in a treatment study of suicidal behavior.
The purpose of this study was to examine patterns of self-reported suicidality and distress during research assessments in a sample of 63 women meeting criteria for borderline personality disorder and current and chronic suicidality. The risk management protocol we used during the two-year study period (University of Washington Risk Assessment Protocol; UWRAP) is described. Results indicated that changes in suicidality following assessments were small and relatively infrequent, and were just as likely to reflect decreases in suicidality as increases (17.5% versus 16.4% of sessions, respectively). Further, longitudinal analyses indicated that changes in suicidality became increasingly rare over the course of the 2-year study. Ratings of distress were more changeable than suicidality, underscoring the need for separate measurement of these constructs when assessing risk. With the aid of the UWRAP, our assessors judged 15 participants as high-risk status in 28 assessment sessions (3.7% of all sessions). In comparison to the rest of the sample, these individuals were of significantly greater clinical severity as measured by the HRSD 17-item, GAF scores, number and severity of previous suicide attempts, and number of previous psychiatric hospitalizations. Low-intensity risk intervention strategies (e.g., validating participant's feelings) were typically sufficient to reduce risk in these participants. Overall, our findings indicate that research with highly suicidal individuals can be done safely with the use of well-trained assessors and an appropriate crisis management protocol.


Nondiscrimination on the basis of handicap; procedures and guidelines relating to health care for handicapped infants--HHS. Final rules.
These are final rules on procedures and guidelines relating to nondiscrimination on the basis of handicap in connection with health care for handicapped infants. These rules are issued under the authority of section 504 of the Rehabilitation Act of 1973, which prohibits discrimination on the basis of handicap in programs and activities receiving Federal financial assistance.


Activation of angiotensin-converting enzyme expression in infarct zone following myocardial infarction.
In the present study we quantified angiotensin-converting enzyme (ACE) mRNA and localized ACE mRNA and protein in the infarcted rat heart. Wistar rats underwent ligation of the left descending coronary artery, resulting in myocardial infarction (MI) or a sham operation. At different times (1-90 days) after surgery (n = 3 each), the heart was removed and divided into the right ventricle (RV), septum (Se) and left ventricle (LV). ACE mRNA was quantified by competitive reverse transcriptase-polymerase chain reaction (RT-PCR). At 4 and 7 days after MI, we found a 2.8-fold increase of ACE mRNA (n = 3; P < or = 0.05) in the infarcted LV compared with the LV of the sham group. No increases of ACE mRNA were found in the noninfarcted hypertrophied compartments. ACE activity increased 2.6- and 3.6-fold in the infarcted LV at 7 and 90 days after MI, respectively. In situ hybridization and immunohistochemistry showed increased ACE mRNA and protein density in the border zone of the infarcted area, predominantly in the endothelial cells lining capillaries. In the noninfarcted myocardium ACE mRNA and protein were confined to endothelial cells of the larger vessels. From these data we conclude that the intracardiac RAS is involved in the healing of the scar after MI in the rat, possibly giving rise to neovascularization. Furthermore, the data suggest that the intracardiac ACE is not necessarily associated with hypertrophy in the rat heart after MI.


[Improvement in the accuracy of diagnosis of ischemic heart disease with simultaneous radioisotope evaluation of regional myocardial function perfusion and an exercise test].
The recently developed myocardial agent methoxy-isobutyl-isonitrile (MIBI), labelled to 99mTc, allows one to evaluate global and regional ventricular function as well as myocardial perfusion by means of a single exercise stress test, significantly increasing diagnostic accuracy for coronary artery disease. Between September 1988 and March 1989, 407 patients with either suspected or already ascertained coronary artery disease underwent simultaneous assessment of regional ventricular function with first pass radionuclide angiography, and of myocardial perfusion with single photon emission computerized tomography, by means of 2 injections of 99mTc-MIBI at rest and at peak of the same exercise test. Out of these patients, 56 (52 men and 4 women, whose mean age was 57 +/- 7 years) underwent coronary angiography within 6 months of radionuclide examination and were included in the study. There were 13 1-vessel, 26 2-vessel and 17 3-vessel disease patients. Thirty-six of them had had a previous (greater than 6 months old) myocardial infarction, on the anterior wall in 16 patients, on the inferior wall in 20. In all patients a computerized bicycle stress test was performed; interruption criteria were ST segment depression greater than or equal to 1 mm in 27 patients (48%), the achievement of a heart rate greater than 85% of maximal age-predicted heart rate in 12 patients (22%) and fatigue in 17 patients (30%). Scintigraphic results were compared to angiographic findings: global sensitivity and specificity of the simultaneous evaluation were 82% and 81%, respectively, vs 95% and 56% of functional results and 85% and 71% of perfusion results, respectively. Regional analysis was also performed, by dividing scintigraphic images into the territories supplied by the 3 main coronary vessels, i.e., left anterior descending artery (LAD), left circumflex artery (LCx) and posterior descending artery (PD). On LAD territory the simultaneous evaluation achieved a sensitivity of 88% and a specificity of 63%, vs 98% and 38% of functional analysis and 88% and 50% of perfusional analysis, respectively. On LCx territory sensitivity and specificity were respectively 71% and 96%, vs 91% and 64% of function, and 77% and 96% of perfusion alone. On PD territory sensitivity was 85%, vs 94% and 91%, respectively; specificity was 73%, vs 55% and 55%, respectively. Combined functional and perfusional analysis achieved a 68% sensitivity in identifying less than or equal to 75% coronary narrowings, and a 90% sensitivity for greater than 90% narrowings.(ABSTRACT TRUNCATED AT 400 WORDS)


Big Data and Health Care: Challenges and Opportunities for Coordinated Policy Development in the EU.
Abstract-As global policy makers prioritize big data policy, it is important to try to outline expected outcomes vis-à-vis health sector objectives. We identify initiatives aimed at promoting the use of big data in European Union (EU) health care, highlight expected challenges, and use these to evaluate EU big data policy developments to the extent that they are able to advance health sector priorities. A comprehensive approach is used to capture and examine peer-reviewed and gray literature publications on the use of big data in global health systems. This approach involved electronic database and specialist website searching, as well as complementary use of search engines and qualitative inputs from key EU policy stakeholders. Ongoing health data initiatives revolve around data center development, confidentiality and security, e-health and m-health, and genomics and bioinformatics. The literature acknowledges several main challenges to the successful integration of big data in health care, classified as either ethical (confidentiality and data security, access to information) or technical (data reliability, interoperability, data management and governance). EU data policy has started to address these issues, though additional work remains. A larger outstanding challenge is the lack of a comprehensive health and research policy strategy for big data that targets sectoral objectives. It remains unclear how big data integration will affect the quality and performance of health care in the EU. The promises of big data are being eroded by a failure to develop a coherent approach to adequately address conceptual, ethical, and technical challenges pertaining to its use within EU health systems.


Establishing LA VIDA: a community-based partnership to prevent intimate violence against Latina women.
LA VIDA--the Southwest Detroit Partnership to Prevent Intimate Violence Against Latina Women--evolved in response to community concern about the problem of intimate partner violence (IPV) and the lack of culturally competent preventive and support services for Latino women and men in southwest Detroit. Since 1997, diverse organizations have mobilized as a community-academic partnership to ensure the availability, accessibility, and utilization of IPV services. This article describes and analyzes the evolution of LA VIDA within a community-based participatory research framework using a case study approach that draws on multiple data sources including group and individual interviews and field notes. The challenges and lessons learned in addressing a complex multifaceted problem such as IPV in an ethnic minority community are highlighted in an examination of the process of mobilizing diverse organizations, conducting community diagnosis and needs assessment activities, establishing goals and objectives within a social ecological framework, and integrating evaluation during the development phase.


A latent class analysis of alcohol abuse and dependence symptoms among Puerto Rican youth.
The study aim was to classify youths into homogenous groups based on their symptoms of Alcohol Abuse and Dependence. An NIDA-funded cross-sectional survey was administered to 622 middle and high school students in San Juan, Puerto Rico in 2000. Latent class analysis (LCA) examined the Alcohol Abuse and Dependence symptoms. Three distinct classes of drinkers were found: low (86.0% of sample), moderate (11.7% of sample), and high (2.3% of sample) severity classes which were distinguished by differing estimated probabilities of symptom reporting. The study has implications for the diagnosis of Alcohol Abuse and Dependence among Puerto Rican youths. Limitations are also noted.


Enhancement of host resistance to viral and tumor challenge by treatment with methionine-enkephalin.
Host resistance to disease is dependent upon a number of factors. Recent evidence indicates that natural killer (NK) cells play an important role in resistance to both neoplastic and virally induced disease. Treatment of C57Bl/6 mice with methionine-enkephalin (1, 3, 10, or 30 mg/kg body weight) results in significant increases in NK activity of splenic lymphocytes 20 hours after injection of the enkephalin. Enkephalin treatment also enhances host resistance. The short-term survival of A/J female mice after HSV-2 infection was significantly increased by daily subcutaneous injections (3 mg/kg body weight) of methionine-enkephalin. Similarly, daily doses of 50 micrograms of methionine-enkephalin for 7 to 14 days inhibit the local subcutaneous tumor growth of B15 melanoma in C57Bl/6 mice.


Characteristics of cultured tomato cells after prolonged exposure to medium containing polyethylene glycol.
Cell lines of tomato (Lycopersicon esculentum Mill., cv. VFNT-Cherry) have been isolated, which are capable of growing in media containing polyethylene glycol (PEG) 6000 with water potentials as low as -15 or -22 bar. After prolonged exposure to media containing PEG, these cell populations have reduced abilities to grow in the absence of PEG. Upon resuspension in PEG-free medium, the cells swell and begin to release metabolites, including protein. Measurement by plasmometry of the osmotic potential of cells selected in medium with -22 bar water potential indicates that they maintain, at the end of the growth cycle, an osmotic potential of approximately -26 bar. This is compared to an osmotic potential of -9 bar for nonselected cells in medium without PEG, having an initial water potential of -4 bar. Thus, considerable osmotic adjustment occurs as a result of exposure to external low water potential. The results also indicate that PEG does not contribute significantly to osmotic adjustment of the cells.


Small Numbers, Big Challenges: Adolescent and Young Adult Cancer Incidence and Survival in New Zealand.
This study was undertaken to determine cancer survival and describe the unique spectrum of cancers diagnosed among New Zealand's adolescents and young adult (AYA) population. Registrations for 1606 15-24 year olds diagnosed with a new primary malignant tumor between 2000 and 2009 were obtained from the New Zealand Cancer Registry and classified according to AYA diagnostic group and subgroup, age, sex, and prioritized ethnicity. Age-standardized incidence rates (IRs) per million person years and 5-year relative survival ratios were calculated. Cancer incidence was 228.6 per million for adolescents aged 15-19 years and 325.7 per million for young adults aged 20-24 years. Overall IRs were consistent across all ethnic groups but there were unique ethnic differences by tumor group including a higher incidence of bone tumors, carcinoma of the gastrointestinal tract, and gonadal germ cell tumors among Maori, a higher incidence of leukemia among Pacific peoples, and a higher incidence of melanoma among non-Maori/non-Pacific peoples. Five-year relative survival for adolescents (75.1%) and AYA overall (80.6%) appeared poorer than had been achieved in other high-income countries. Maori (69.5%) and Pacific (71.3%) AYA had lower 5-year survival compared to non-Maori/non-Pacific peoples (84.2%). The survival disparities observed require further investigation to identify and address the causes of these inferior outcomes. The newly established AYA Cancer Network Aotearoa has been tasked with improving cancer survival and care and ensuring equality of access for New Zealand AYAs with cancer.


beta-Adrenergic signaling and thyroid hormones affect HSP72 expression during heat acclimation.
Heat acclimation upregulates 72-kDa heat shock protein (HSP72) and predisposes to faster activation of the heat shock response (HSR). This study investigates the role played by beta-adrenergic signaling and/or plasma thyroxine level in eliciting these features by using rats undergoing 1) heat acclimation (AC; 34 degrees C, 2 and 30 days); 2) AC with beta-adrenergic blockade; 3) AC-maintained euthyroid; 4) hypothyroid; 5) hyperthyroid; and 6) controls. The hsp72 mRNA (RT-PCR) and HSP72 levels (Western blot) were measured before and after heat stress (2 h, 41 degrees C, rectal temperature monitored). beta-Adrenergic blockade during AC abolished HSP72 accumulation, without disrupting HSR. Low thyroxine blunted the HSR at posttranscriptional level, whereas thyroxine administration in hyperthyroid and AC-maintained euthyroid rats arrested heat stress-evoked hsp72 transcription. We conclude that beta-adrenergic signaling contributes to the high HSP72 level characterizing the AC state. Thyroxine has two opposing effects: 1) direct repressive on rapid hsp72 transcription after heat stress; and 2) indirect stimulatory via beta-adrenergic signaling. Low thyroxine could account for diminished HSP72 synthesis via lower heat production and thermoregulatory set point.


A comparison of calcium homeostasis in isolated and attached growth cones of the snail Helisoma.
This study examines the capability of growth cones from identified neurons of the snail Helisoma trivolvis to perform calcium homeostasis. Calcium influx into the cytoplasm was eliminated or increased experimentally to alter [Ca]i, and the compensatory response of the growth cone was measured with the fluorescent calcium indicator Fura-2. Growth cones compensated for both increases and decreases in calcium influx by restoring [Ca]i towards basal levels under both types of challenges. The intrinsic ability of growth cones to control [Ca]i was examined in physically isolated growth cones. Isolated growth cones demonstrated essentially identical calcium homeostatic properties to their intact counterparts, indicating that mechanisms governing calcium homeostasis exist intrinsically in the growth cone. Such independence may add significantly to the growth cone's potential to locally interpret and respond to stimuli encountered en route to its appropriate target.


Highly Selective Enzymatic Recovery of Building Blocks from Wool-Cotton-Polyester Textile Waste Blends.
In Europe, most of the discarded and un-wearable textiles are incinerated or landfilled. In this study, we present an enzyme-based strategy for the recovery of valuable building blocks from mixed textile waste and blends as a circular economy concept. Therefore, model and real textile waste were sequentially incubated with (1) protease for the extraction of amino acids from wool components (95% efficiency) and (2) cellulases for the recovery of glucose from cotton and rayon constituents (85% efficiency). The purity of the remaining poly(ethylene terephthalate) (PET) unaltered by the enzymatic treatments was assessed via Fourier-transformed infrared spectroscopy. Amino acids recovered from wool were characterized via elementary and molecular size analysis, while the glucose resulting from the cotton hydrolysis was successfully converted into ethanol by fermentation with Saccharomyces cerevisiae. This work demonstrated that the step-wise application of enzymes can be used for the recovery of pure building blocks (glucose) and their further reuse in fermentative processes.


Spacial Score─A Comprehensive Topological Indicator for Small-Molecule Complexity.
The fraction of sp3-hybridized carbons (Fsp3) and the fraction of stereogenic carbons (FCstereo) are two widely employed scores of molecular complexity with strong links to biologically relevant features. However, they do not comprehensively express molecular topology, and they often do not match the chemical intuition of complexity. We propose the spacial score (SPS) as an empirical scoring system that builds upon the principle underlying Fsp3 and FCstereo and expresses the spacial complexity of a compound in a uniform manner on a highly granular scale. The size-normalized SPS (nSPS) can differentiate distributions of natural products and synthetic compounds and is applicable in the analysis of biological activity data. Analysis of the ChEMBL database revealed general trends of increasing selectivity and potency with increasing nSPS. SPS can also be used advantageously in planning and analysis of synthesis programs for direct comparison of chemical transformations and intermediates in reaction sequences.


Pharmacokinetics of phenytoin following intravenous and intramuscular administration of fosphenytoin and phenytoin sodium in the rabbit.
The purpose of this study was to evaluate and compare plasma phenytoin concentration versus time profiles following intravenous (i.v.) and intramuscular (i.m.) administration of fosphenytoin sodium with those obtained following administration of standard phenytoin sodium injection in the rabbit. Twenty-four adult New Zealand White rabbits (2.1 +/- 0.4 kg) were anaesthetized with sodium pentobarbitone (30 mg/kg) followed by i.v. or i.m. administration of a single 10 mg/kg phenytoin sodium or fosphenytoin sodium equivalents. Blood samples (1.5 ml) were obtained from a femoral artery cannula predose and at 1, 3, 5, 7, 10, 15, 20, 30, 45, 60, 90, 120, 180, 240 and 300 min after drug administration. Plasma was separated by centrifugation (1000 g; 5 min) and fosphenytoin, total and free plasma phenytoin concentrations were measured using high performance liquid chromatography (HPLC). Following i.v. administration of fosphenytoin sodium plasma phenytoin concentrations were similar to those obtained following i.v. administration of an equivalent dose of phenytoin sodium. Mean peak plasma phenytoin concentrations (Cmax) was 158% higher (P = 0.0277) following i.m. administration of fosphenytoin sodium compared to i.m. administration of phenytoin sodium. The mean area under the plasma total and free phenytoin concentration-time curve from time zero to 120 min (AUC(0-120)) following i.m. administration was also significantly higher (P = 0.0277) in fosphenytoin treated rabbits compared to the phenytoin group. However, there was no significant difference in AUC(0-180) between fosphenytoin and phenytoin-treated rabbits following i.v. administration. There was also no significant difference in the mean times to achieve peak plasma phenytoin concentrations (Tmax) between fosphenytoin and phenytoin-treated rabbits following i.m. administration. Mean plasma albumin concentrations were comparable in both groups of animals. Fosphenytoin was rapidly converted to phenytoin both after i.v. and i.m. administration, with plasma fosphenytoin concentrations declining rapidly to undetectable levels within 10 min following administration via either route. These results confirm the rapid and complete hydrolysis of fosphenytoin to phenytoin in vivo, and the potential of the i.m. route for administration of fosphenytoin delivering phenytoin in clinical settings where i.v. administration may not be feasible.


Response of refractory colitis to intravenous or oral tacrolimus (FK506).
Intravenous cyclosporine has proven to be an alternative to emergency colectomy in steroid-refractory ulcerative colitis, whereas the experience with FK506 is limited. In this report we compare intravenous to oral FK506 treatment in 38 patients with refractory ulcerative (n = 33) or indeterminate (n = 5) colitis. FK506 was started intravenously in the first group (n = 18) at a dose of 0.01 to 0.02 mg/kg up to 14 days, followed by 0.1 to 0.2 mg/kg orally, or was started orally at this dose in a second group (n = 20). Additional azathioprine/6-mercaptopurine was given and steroids were tapered in responding patients, followed by a dose reduction of FK506. Clinical disease activity and laboratory parameters were assessed to evaluate efficacy and safety. Primary objectives were the induction of remission (Truelove index of mild) and colectomy-free survival. Treatment lasted for a mean of 7.6 months, and the mean observation period was 16.2 months. Eighteen of 38 patients improved within 14 days, and a complete remission was achieved in 13 patients after 1 month. A colectomy within 1 month was performed in 3 of 38 patients. The overall colectomy rate was 34%. One-half of the patients with a minimum follow-up of 2 years required a colectomy. Intravenous and per oral administration were equally safe and effective. The most frequent adverse events included tremor, hyperglycemia, hypertension, and infection, but none were severe. Renal impairment was rare and subsided upon drug withdrawal. In conclusion. FK506 is effective in the treatment of refractory colitis with per oral dosing being equivalent to intravenous administration.


Bisphenol A from dental polycarbonate crown upregulates the expression of hTERT.
Bisphenol A (BPA) is one of the endocrine-disrupting chemicals (EDCs) that possess estrogen-like biologic activity. Many dental materials have been reported to release BPA. However, there are few reports available on the release of BPA from dental polycarbonates. The purpose of this study was to investigate the release of BPA from dental polycarbonate crowns and to evaluate the estrogenic activity of BPA. Polycarbonate crowns were immersed in five solvents (water, ethanol, n-hepthane, acetic acid, and acetonitrile) at 37 or 65 degrees C for 24 h. The elution from the material was analyzed by high-performance liquid-chromatography (HPLC) and mass-spectrometry (MS) analysis. BPA release was detected corresponding to the degradation of dental polycarbonates under the some storage conditions (ethanol, acetic acid, and acetonitrile). A previous report proved that estrogen increased human telomerase catalytic subunit (hTERT) mRNA, whereas the effect of EDCs on the hTERT promoter has never been reported. The estrogenic activity of BPA was analyzed by luciferase assay with the use of the hTERT promoter. This assay revealed that BPA was a positive regulator of hTERT transcription. In addition, quantitative real-time PCR analysis showed that BPA increased the expression level of hTERT mRNA in MCF7 cells. Herein, it is demonstrated that hTERT is a new target of BPA.


Interference of psychotropic drugs with the injuring action of ethanol on gastric mucosal barrier.
The aim of this study was to establish whether some psychotropic drugs, applied to patients under influence of alcohol, may potentiate its damaging action on gastric mucosal barrier. A sensitive index of such damage is a decline of the potential difference (PD) across the stomach wall. The experiments were carried out on Wistar rats of either sex, anesthetized with urethane-chloralose. The PD values were assayed with an apolarization method. The investigated solutions were administered intragastrically by gavage. Ethanol at a concentration of 40% v/v depressed PD by 39%. The investigated psychotropic drugs did not change PD by themselves but given in combination with ethanol caused significant decline of PD: diazepam (0.4 mg/kg) by 58%, chlorpromazine (6.7 mg/kg) by 59%, imipramine (2 mg/kg) by 48%, amitriptyline (4 mg/kg) by 49%, phenytoin (4 mg/kg) by 53%, pyridinol (0.3 mg/kg) by 58%. Intragastric administration of water did not change PD. The results indicate that while psychotropic drugs given alone do not affect significantly the gastric mucosal barrier, they may potentiate the damaging action of ethanol on this barrier.


A fatal Sunday brunch: Amanita mushroom poisoning in a Gulf Coast family.
Mushroom poisoning with resultant liver toxicity often occurs in experienced collectors. The increasing popularity of natural foods has lead to an increased incidence of mushroom poisoning among the general population. This report describes a grandmother with over 30 yr of self-taught expertise in selecting and preparing wild mushrooms, who prepared a mushroom dish that led to the poisoning of herself and three family members. A 3-yr-old child died of fulminant hepatic failure, and the others recovered completely. The clinical presentation, course, and management of patients with mushroom poisoning are discussed.


Delayed coma in head injury: consider cerebral fat embolism.
To describe a case of a young man with delayed coma after mild head injury, suggestive of cerebral fat embolism (CFE). To underline the value of MR imaging in the differential diagnosis of secondary deterioration in mild head injury. A 21-year-old man admitted with mild head injury after a fall with facial fractures and long bone fractures. He was admitted to the intensive care unit and was mechanically ventilated. Weaning was not possible because of desaturations and pulmonary congestion. Low platelet count and anaemia developed. On several time points during his admission cerebral imaging data were obtained. Non-contrast CT on admission was normal while follow-up MRI showed extensive white matter abnormalities. These imaging abnormalities combined with the clinical presentation suggests cerebral fat embolism (CFE) as the most likely cause of secondary deterioration in our patient. In head injured patients with long bone fractures one should consider cerebral fat embolism. When the classical clinical syndrome is not present, MR imaging is warranted for diagnosis and to exclude other causes of secondary deterioration.


Why is testing for HIV so different?
An HIV test requires counselling, confidentiality and consent. The patient, the doctor or a third party may initiate it. Assessment of risk behaviours, rather than stereotyping into risk groups, is needed. Patients initiate tests after perceived risk behaviour, and other factors such as guilt or anxiety may prompt testing. Clinical cues such as fevers, weight loss, lymphadenopathy and candidiasis should alert general practitioners to possible HIV infection. General practitioners have a pivotal role in HIV testing and management.


Mesoporous Silica Nanocarriers with Cyclic Peptide Gatekeeper: Specific Targeting of Aminopeptidase N and Triggered Drug Release by Stimuli-Responsive Conformational Transformation.
Utilizing stimuli-responsive conformational transformation of a cyclic peptide as a gatekeeper for mesoporous nanocarriers has several advantages such as facile introduction of targeting capabilities, low enzymatic degradation during blood circulation and enhanced specific binding to selected cells. In this report, a Asn-Gly-Arg (NGR)-containing dual-functional cyclic peptide gatekeeper on the surface of mesoporous nanocarrier is prepared not only for active targeting of the aminopeptidase N (APN) expressed on cancer cells but also stimuli-responsive intracellular drug release triggered by a glutathione (GSH)-induced conformational transformation of the peptide gatekeeper. The peptide gatekeeper on the surface of nanocarriers exhibits on-off gatekeeping by conformational transformation triggered by intracellular glutathione of the cancer cells. H1299 cells (high APN expression) showed greater uptake of the nanocarrier by endocytosis and higher apoptosis than A549 cells (low APN expression).


Preterm labor and bacterial intra-amniotic infection: arachidonic acid liberation by phospholipase A2 of Prevotella bivia.
There is a strong association between preterm labor and infection, presumably through an increase in prostaglandin formation. The studies presented in this report were undertaken to evaluate whether Prevotella bivia, a common anaerobic isolate of intrauterine infection, stimulates arachidonic acid metabolism, as a rate-limiting step for prostaglandin synthesis in the human uterine endometrium. When human uterine endometrial cells prelabeled with [3H]arachidonic acid to an isotopically steady state were exposed to an extract of P. bivia, arachidonic acid liberation was stimulated, accompanied by lysophospholipid formation. Similar stimulatory effect on phospholipid degradation was also observed in the experiment with the bacterial conditioned media which was spent as culture media. These results suggests that P. bivia stimulates endometrial phospholipid metabolism, related with activity of phospholipase A2, which might induce the onset of labor associated with intra-amniotic infection.


cDNA cloning and functional expression in yeast Saccharomyces cerevisiae of beta-naphthoflavone-induced rabbit liver P-450 LM4 and LM6.
A cDNA library was constructed from liver mRNA of a beta-naphthoflavone-induced rabbit. Two clones pLM4-1 and pLM6-1 containing 2.2-kbp inserts that hybridized at low stringincy with a mouse P1 P-450 probe were selected. The clone pLM4-1 was fully sequenced and found to contain a full-length cDNA coding for cytochrome P-450 LM4. Partial sequence and restriction mapping made it possible to identify pLM6-1 as coding for the major part of cytochrome P-450 LM6. Cloned LM4-1 cDNA was reformed by deletion of the 5' and 3' non-coding regions before insertion into yeast expression vectors PYe DP1/10. A similar operation was performed on pLM6-1 cDNA after replacement of the missing N-terminus-coding sequences by homologous sequences form the pLM4-1 clone resulting in a chimeric cytochrome P-450 coding sequence. Expression of cloned rabbit cytochrome P-450 into transformed yeast was optimized by studying the effect of the nature of the DNA sequence just preceding the initiation codon on the level of cytochrome P-450 production. Yeast synthesized cytochromes P-450 were characterized by immunoblotting, spectra and catalytic activity determinations. Cloned cytochrome P-450 LM4 was found by all criteria to be identical to the authentic rabbit one. The chimeric cytochrome P-450 that contains the 143 N-terminal amino acids of cytochrome P-450 LM4 and the remaining 375 amino acids of cytochrome P-450 LM6 was found to exhibit most of the authentic cytochrome P-450 LM6 catalytic properties. Enzymatic and evolutionary implications of these results are discussed.


Influenza A Virus as a Predisposing Factor for Cryptococcosis.
Influenza A virus (IAV) infects millions of people annually and predisposes to secondary bacterial infections. Inhalation of fungi within the Cryptococcus complex causes pulmonary disease with secondary meningo-encephalitis. Underlying pulmonary disease is a strong risk factor for development of C. gattii cryptococcosis though the effect of concurrent infection with IAV has not been studied. We developed an in vivo model of Influenza A H1N1 and C. gattii co-infection. Co-infection resulted in a major increase in morbidity and mortality, with severe lung damage and a high brain fungal burden when mice were infected in the acute phase of influenza multiplication. Furthermore, IAV alters the host response to C. gattii, leading to recruitment of significantly more neutrophils and macrophages into the lungs. Moreover, IAV induced the production of type 1 interferons (IFN-α4/β) and the levels of IFN-γ were significantly reduced, which can be associated with impairment of the immune response to Cryptococcus during co-infection. Phagocytosis, killing of cryptococci and production of reactive oxygen species (ROS) by IAV-infected macrophages were reduced, independent of previous IFN-γ stimulation, leading to increased proliferation of the fungus within macrophages. In conclusion, IAV infection is a predisposing factor for severe disease and adverse outcomes in mice co-infected with C. gattii.


Evaluation of Retinal Ganglion Cell Layer Thickness in the Early Period After Femtosecond LASIK Surgery
To evaluate the early effects of femtosecond laser-assisted in situ keratomileusis (LASIK) surgery on retinal ganglion cell thickness (GCT), peripapillary retinal nerve fiber thickness (NFT), and central macular thickness (CMT) obtained by spectral domain optical coherence tomography (SD-OCT) in a healthy population. This case-control study included data from the right eye of 40 subjects without any disease other than refractive error and who had undergone femtosecond LASIK surgery. The preoperative, postoperative 1-hour, and postoperative 3-week GCT, NFT, and CMT values obtained by SD-OCT were compared. The mean age was 27.54±5.99 years (18-45 years). GCT, NFT, and CMT were 18.43±6.03 μm, 107.90±9.01, and 234.3±21.2 μm preoperatively; 18.05±5.93 μm, 108.08±8.92 μm, and 230.1±22.6 μm at postoperative 1 hour; and 17.86±5.27 μm, 107.98±10.13, and 236.3±25.1 μm at postoperative 3 weeks (p=0.159, 0.85, and 0.254, respectively). There were no changes in GCT, NFT, and CMT values evaluated with SD-OCT in the early period after femtosecond LASIK surgery.


A re-assessment of gene-tag classification approaches for describing var gene expression patterns during human Plasmodium falciparum malaria parasite infections.
PfEMP1 are variant parasite antigens that are inserted on the surface of Plasmodium falciparum infected erythrocytes (IE). Through interactions with various host molecules, PfEMP1 mediate IE sequestration in tissues and play a key role in the pathology of severe malaria. PfEMP1 is encoded by a diverse multi-gene family called var. Previous studies have shown that that expression of specific subsets of var genes are associated with low levels of host immunity and severe malaria. However, in most clinical studies to date, full-length var gene sequences were unavailable and various approaches have been used to make comparisons between var gene expression profiles in different parasite isolates using limited information. Several studies have relied on the classification of a 300 - 500 base-pair "DBLα tag" region in the DBLα domain located at the 5' end of most var genes. We assessed the relationship between various DBLα tag classification methods, and sequence features that are only fully assessable through full-length var gene sequences. We compared these different sequence features in full-length var gene from six fully sequenced laboratory isolates. These comparisons show that despite a long history of recombination, DBLα sequence tag classification can provide functional information on important features of full-length var genes. Notably, a specific subset of DBLα tags previously defined as "group A-like" is associated with CIDRα1 domains proposed to bind to endothelial protein C receptor. This analysis helps to bring together different sources of data that have been used to assess var gene expression in clinical parasite isolates.


In vivo and in vitro inhibition of human liver cancer progress by downregulation of the μ-opioid receptor and relevant mechanisms.
Opiates have long been used as analgesics to relieve pain associated with various medical conditions. μ-opioid receptor (MOR) is the main member of the opioid receptor super-family and the excitation or overexpression of MOR promotes the proliferation of many kinds of tumor cells. It was found in our previous studies that MOR was highly expressed in the tissue and cells of human liver cancer. However, the impact of MOR on the progress of human liver cancer remains unknown. The purpose of this study is to investigate the impact of MOR downregulation on the progress of human liver cancer and the mechanisms involved. RNA interfering or specific inhibitor was administered to downregulate the MOR in human hepato-cellular carcinoma cells and it was found that the proliferation of hepatocellular carcinoma cells was significantly inhibited with the increase of the apoptotic rate, while the cell cycle was blocked in G0/G1 phase and the tumor growth in the mice was retarded. In addition, downregulation of MOR resulted in the increase of phosphorylation of the MKK7 expression and JNK activation. On the contrary, blockade of MKK7 pathway can reverse the antitumor role of MOR. In summary, downregulation of MOR is able to inhibit both in vivo and in vitro human liver cancer progress and it shows potential to be used in cancer therapy.


Comparison of clinical and angiographic features and longterm follow-up events between patients with variant angina and patients with ST elevation myocardial infarction.
We investigated to what extent patients with variant angina and significant coronary stenosis (>or=70%) present a clinical and angiographic profile similar to patients with ST elevation myocardial infarction. Thus, the clinical and angiographic features as well as follow-up events of 200 patients were prospectively analyzed and were compared with those of 422 patients with a first ST elevation myocardial infarction survivors of the early phase (3 days) and those of 70 patients with variant angina and non significant stenosis. Age and incidence of smoking, systemic hypertension, diabetes and maximum ST elevation were similar in the 2 groups. Furthermore, among patients with significant coronary stenosis, stenosis severity and the proportion of eccentric lesions were also comparable. Incidence of recent-within 30 days prior to admission-angina at rest was higher in variant angina patients with significant stenosis (67% vs. 27%, p<0.001) than in those with myocardial infarction but long standing angina at rest (>30 days) was low and comparable in these 2 groups (15% vs. 11%, ns). Also, in a 5-year follow-up most patients from these 2 groups were free from angina at rest (86% vs. 84%) which in variant angina patients was largely attributable to a high revascularization rate (72%). Moreover, the rate of myocardial infarction/cardiac death (20% vs. 19%) was also similar. Patients with variant angina and non-significant stenosis, however, had longer antecedent angina, more frequent follow-up angina and a lower incidence of cardiac events than the other 2 groups. Thus, these findings suggest that patients with variant angina and significant coronary stenosis generally behave as an acute coronary syndrome-likely associated with an acutely complicated plaque-rather than as recurrent vasospastic angina, and should be managed accordingly.


Capillary Condensation with a Grain of Salt.
Capillary condensation (CC), namely, the formation from the vapor of a stable phase of drops below the saturation pressure, is a prevalent phenomenon. It may occur inside porous structures or between surfaces of particles. CC between surfaces, a liquid "bridge", is of particular practical interest because of its resulting adhesive force. To date, studies have focused on pure water condensation. However, nonvolatile materials, such as salts and surfactants, are prevalent in many environments. In the current study, the effect of these contaminants on CC is investigated from a thermodynamic point of view. This is done by computing the Gibbs energy of such systems and developing the modified Kelvin equation, based on the Kohler theory. The results demonstrate that nonvolatile solutes may have a number of major effects, including an increase in the critical radius and the stabilization of the newly formed phase.


Ultraviolet polarization vision and visually guided behavior in fishes.
Teleost fishes are capable of detecting and behaviorally responding to linearly polarized light. Fish exhibit free-swimming spatial orientation to imposed and natural polarized light fields, and the fidelity of this spatial orientation depends heavily on UV and short wavelength content of the polarization field. Fish make fine-scale behavioral discriminations between stimuli that differ in e-vector orientation, independent of brightness. The detection of polarized light by photoreceptors is based on specializations of the disk membrane in the outer segment of cones that permit preferential absorption of axial and transverse polarized light. Differential polarization detectors that have overlapping spectral sensitivity in the UV short wavelength spectrum mediate polarization sensitivity. These differential detectors are based on cone photoreceptors that share spectral sensitivity in the UV short wavelength spectrum: the alpha-band of UV-sensitive cone mechanism as the vertical detector, and the beta-band of mid- and long-wavelength sensitive cone mechanisms as the horizontal detector. Negative feedback of horizontal cells on cones govern opponent interactions between differentially sensitive polarization detectors. Polarization opponency functions to enhance e-vector contrast under conditions that vary in degree of polarization and ambient intensity. Ontogenetic changes in the cone mosaic, resulting from programmed cell death and regeneration of UV-sensitive cones, alter the retinal location of polarization sensitivity. These developmental changes greatly influence behavioral responses to polarized light.


Effect of a New Hyaluronic Acid Hydrogel Dermal Filler Cross-Linked With Lysine Amino Acid for Skin Augmentation and Rejuvenation.
Hyaluronic acid (HA) fillers are the most popular filler agents for skin rejuvenation. Although 1,4-butanediol diglycidyl ether is regarded as a relatively safe cross-linker, it still exhibits certain cytotoxicity. We presented here an amino acid-cross-linked HA (ACHA) which was obtained by an amidation reaction with lysine and HA. This study aimed to investigate ACHA's efficacy and safety for skin augmentation and rejuvenation. Rheology, compressive tests, and swelling experiments were conducted to investigate ACHA's mechanical and viscoelastic properties. The effects of ACHA on the human keratinocytes (HaCaT) cells and the human dermal fibroblast (HDF) were investigated by Transwell and wound healing assays. Its impacts on the epithelial thickness and collagen synthesis were further examined in a mouse experimental model. We recruited 50 patients with moderate to severe nasolabial folds (NLFs). The patients were randomly allocated to receive ACHA or Restylane injections. The resulting retention rates of HA and the Wrinkle Severity Rating Scale and Global Aesthetic Improvement Scale outcomes were evaluated and compared. ACHA exhibited good viscoelasticity. It not only promoted migration and proliferation of HaCat and HDF and secretion of various growth factors but also increased skin thickness and promoted the generation of collagen. Patients who received ACHA had more residual volume 12 months after treatment. ACHA exhibited a promising augmentation effect in NLF correction with few adverse reactions. ACHA has shown promise as a biomaterial with excellent biocompatibility and viscoelastic characteristics in both research and the clinic.See the abstract translated into Hindi, Portuguese, Korean, German, Italian, Arabic, Chinese, and Taiwanese online here: https://doi.org/10.1093/asj/sjad169.


The impact of circumstances surrounding the death of a child on parents' grief.
A longitudinal study was conducted among bereaved parents to examine the relationship between the circumstances surrounding the death of their child and psychological adjustment. Two hundred nineteen couples participated at 6, 13, and 20 months post-loss. Examination was made of two categories of factors: those that were determined by the particular death circumstances (e.g., whether the parent was present at the death) versus those over which parents themselves could have influence (e.g., choice of cremation or burial). Results indicated that some but not all factors were related to adjustment over time. Importantly, the feeling of having said goodbye to the child and presenting the body for viewing at home were associated with lower levels of the parents' grief. Implications for supporting bereaved parents are discussed.


Thyroid fine-needle aspiration cytology in Taiwan: a nationwide survey and literature update.
In Taiwan, thyroid fine-needle aspiration cytology is easily accessible and reliable for evaluating thyroid nodules. The sonographic pattern plays a major role and is the deciding factor for aspiration. We conducted a nationwide survey in 2017 and it revealed that 31% of laboratories had adopted The Bethesda System for Reporting Thyroid Cytopathology. There was a relatively high unsatisfactory rate (24.04%) and low rates of indeterminate diagnoses, including atypia of undetermined significance/follicular lesions of undetermined significance: 4.87%, and follicular neoplasm/suspicious for a follicular neoplasm: 0.35%. Moreover, the risks of malignancy in benign, atypia of undetermined significance, and suspicious for a follicular neoplasm were relatively high. These may reflect strict diagnostic criteria for indeterminate categories and better patient selection for surgery. Improvements in specimen sampling and continuing education programs are crucial. Newly-developed thyroid cytology technologies, such as immunocytochemistry, molecular testing, and computerized cytomorphometry, may further facilitate cytology diagnoses.


Minimal oligosaccharide structures required for induction of immune responses against meningococcal immunotype L1, L2, and L3,7,9 lipopolysaccharides determined by using synthetic oligosaccharide-protein conjugates.
The 12 types of meningococcal lipopolysaccharide (LPS) (immunotypes) contain immunotype-specific and cross-reactive epitopes situated on the oligosaccharide part of the LPS molecules. To identify useful cross-reactive epitopes and to determine minimal oligosaccharide structures required for the induction of an immune response against the most prevalent immunotypes, L1, L2, and L3,7,9, synthetic as well as native LPS-derived oligosaccharides were conjugated with tetanus toxoid. L3,7,9 phosphoethanolamine (PEA) group-containing oligosaccharide-tetanus toxoid conjugates evoked high immunoglobulin G (IgG) antibody levels in rabbits which were detected by an L2-, L3,7,9-, and, depending on the antiserum, L1-specific enzyme-linked immunosorbent assay (ELISA). Inhibition studies revealed that an identical antibody population was detected by L1 and L3,7,9 ELISA, indicating a similar tertiary structure of the inner core oligosaccharide of these two immunotypes. These antibodies recognize PEA group-containing epitopes present on the L1 and L3,7,9 LPS. An L2 PEA group-containing oligosaccharide-tetanus toxoid conjugate elicited L2- and L3,7,9-specific IgG antibodies, but in contrast with the L3,7,9 conjugates, no L1-specific IgG antibodies were evoked. These results indicate that L1 and L2 LPS do not contain cross-reactive epitopes, whereas both L2 and L3,7,9 LPS and L1 and L3,7,9 LPS possess common determinants. Three linear oligosaccharides and one branched oligosaccharide, representing partial structures of the inner core oligosacchardes of meningococcal LPS, were synthesized. Only the branched synthetic oligosaccharide-containing conjugate was able to induce and L1- and L3,7,9-specific immune response, whereas the linear oligosaccharide-protein conjugates evoked L2-specific immune responses. The branched oligosaccharide (beta-D-Glcp(1----4)-[L-alpha-D-Hepp(1----3)]-L-alpha-D-Hepp ) is therefore considered a minimal structure required for the induction of an immune response against L1 and L3,7,9 LPS and part of a cross-reactive epitope between these two immunotypes. For L2-specific immune responses, oligosaccharide structures terminating in beta-D-Glcp(1----4), alpha-D-GlcNAcp(1----2), or L-alpha-D-Hepp(1----5) are needed. The results suggest that it is possible to prepare an oligosaccharide structure with the ability to evoke an immune response against L1, L2, and L3,7,9 LPS. A feasible structure for such a "hybrid" oligosaccharide is discussed.


Veteran and Provider Satisfaction with a Home-Based Telerehabilitation Assessment for Wheelchair Seating and Mobility.
The objective of this project was to measure Veteran and provider satisfaction with a home-based telerehabilitation assessment for wheelchair seating and mobility. Forty-three Veterans were seen remotely at their place of residence by a provider, using a VA Video Connect synchronous videoconferencing system. Veteran and provider satisfaction were collected using the Telerehabilitation Questionnaire (TRQ). Mean individual TRQ scores for both Veterans and providers were significantly higher than the scale midpoint of 3.5. Veterans had higher scores than providers for five individual items on the TRQ. Higher scores by Veterans on the technology and quality and clarity of the video and audio likely correspond to the differences in environmental settings in which the visit occurred for the Veteran compared with the provider. High satisfaction scores with the telerehabilitation assessments are likely attributed to the positive working relationship between the provider and the rehabilitation technician, who provided in-person technical support to the Veteran in the home during the wheeled mobility evaluation. Overall, the results indicate a high level of Veteran and provider satisfaction using telerehabilitation for wheelchair seating and mobility evaluations.


[Diagnostic imaging methods in neonates with diseases of the urinary tract].
The actually used examination algorithm of imaging methods was checked in 35 neonates examined in the course of two years (1991 and 1992) on account of suspected diseases of the urinary pathways. It involved sonographic examination of 28 children, voiding cystourethrography (34x) and intravenous urography (18x). Ascendent pyelography and irrigography were performed three times, one neonate was examined by computed tomography. The procedure used as a basic sonographic prenatal screening in 11 children, in 20 clinical suspicion of disease of the urinary pathways, and four neonates were examined on account of an anorectal malformation or abnormality of the external genitalis. The most frequent finding--in 18 children--was hydronephrosis. It was associated with signs of subrenal obstruction (8x), with ren duplex (5x) and with a megaureter (6x). The association of the sonographic finding of dilatation of the intrarenal urinary pathways with absent contrast filling during excretory urography was observed in a child with hydronephrosis and subrenal obstruction and in four children with a multicystic dysplastic kidney. In individual cases the authors detected a valve of the posterior urethra, a sigmoid-vesical fistula, agenesis of the penis and stenosis of the urethra. Eleven neonates had a normal findings of the urinary tract.


Failure to recruit anti-inflammatory CD103+ dendritic cells and a diminished CD4+ Foxp3+ regulatory T cell pool in mice that display excessive lung inflammation and increased susceptibility to Mycobacterium tuberculosis.
Susceptibility to Mycobacterium tuberculosis is characterized by excessive lung inflammation, tissue damage, and failure to control bacterial growth. To increase our understanding of mechanisms that may regulate the host immune response in the lungs, we characterized dendritic cells expressing CD103 (α(E) integrin) (αE-DCs) and CD4(+) Foxp3(+) regulatory T (T(reg)) cells during M. tuberculosis infection. In resistant C57BL/6 and BALB/c mice, the number of lung αE-DCs increased dramatically during M. tuberculosis infection. In contrast, highly susceptible DBA/2 mice failed to recruit αE-DCs even during chronic infection. Even though tumor necrosis factor alpha (TNF-α) is produced by multiple DCs and macrophage subsets and is required for control of bacterial growth, αE-DCs remained TNF-α negative. Instead, αE-DCs contained a high number of transforming growth factor beta-producing cells in infected mice. Further, we show that T(reg) cells in C57BL/6 and DBA/2 mice induce gamma interferon during pulmonary tuberculosis. In contrast to resistant mice, the T(reg) cell population was diminished in the lungs, but not in the draining pulmonary lymph nodes (PLN), of highly susceptible mice during chronic infection. T(reg) cells have been reported to inhibit M. tuberculosis-specific T cell immunity, leading to increased bacterial growth. Still, despite the reduced number of lung T(reg) cells in DBA/2 mice, the bacterial load in the lungs was increased compared to resistant animals. Our results show that αE-DCs and T(reg) cells that may regulate the host immune response are increased in M. tuberculosis-infected lungs of resistant mice but diminished in infected lungs of susceptible mice.


Competence of Immature Maize Embryos for Agrobacterium-Mediated Gene Transfer.
Agrobacterium-mediated transfer of viral sequences to plant cells (agroinfection) was applied to study the susceptibility of immature maize embryos to the pathogen. The shoot apical meristem of immature embryos 10 to 20 days after pollination from four different maize genotypes was investigated for competence for agroinfection. There was a direct correlation between different morphological stages of the unwounded immature embryos and their competence for agroinfection. Agroinfection frequency was highest in the embryogenic line A188. All developmental stages tested showed Agrobacterium virulence gene-inducing activity, whereas bacteriocidal substances were produced at stages of the immature embryos competent for agroinfection. The results suggested that Agrobacterium may require differentiated tissue in the maize shoot apical meristem before wounding for successful T-DNA transfer. This requirement for the young maize embryo has implications for the possible use of Agrobacterium for maize transformation.


Do you feel happy when other members look happy? Moderating effect of community-level social capital on interconnection of happiness.
One's happiness is expected to be affected by the happiness of surrounding others. This socio-psychological nature of happiness, however, has not been fully examined in the literature of social psychology. The current study examined if this "psychological interconnection of happiness" occurs when (i) individuals have strong personal social capital and/or (ii) individuals belong to a community where other members have strong social capital. We analysed a large social survey dataset sampled from 408 communities in Japan (N = 7295). The psychological interconnection of happiness was measured by calculating the correlation between individual happiness and perceived community happiness. The multilevel analyses revealed that the psychological interconnection of happiness was moderated by community-level social capital above and beyond individual-level social capital, while individual-level social capital did not have a significant moderation effect.


Supplementation with omega-3 polyunsaturated fatty acids in the management of dysmenorrhea in adolescents.
The purpose of the study was to examine whether dietary supplementation with omega-3 fatty acids can relieve symptoms of dysmenorrhea in adolescents. Forty-two adolescents with dysmenorrhea were randomly allocated to two groups. In the first group 21 girls received fish oil (1080 mg icosapentaenoic acid, 720 mg docosahexaenoic acid, and 1.5 mg vitamin E) daily for 2 months followed by a placebo for an additional 2 months. In the second group 21 girls received placebo for the first 2 months, followed by fish oil for 2 more months. The Cox Menstrual Symptom Scale was used to assess response to treatment. There were no significant differences in the Cox Menstrual Symptom Scale between the two groups at baseline after 2 months of placebo administration. After 2 months of treatment with fish oil there was a marked reduction in the Cox Menstrual Symptom Scale from a baseline mean value of 69.9 to 44.0 (p < 0.0004). This study suggests that dietary supplementation with omega-3 fatty acids has a beneficial effect on symptoms of dysmenorrhea in adolescents.


Wave Reflection and Ventriculo-Arterial Coupling in Bicuspid Aortic Valve Patients With Repaired Aortic Coarctation.
Ventriculo-arterial (VA) coupling in bicuspid aortic valve (BAV) patients can be affected by the global aortopathy characterizing BAV disease and the presence of concomitant congenital lesions such as aortic coarctation (COA). This study aimed to isolate the COA variable and use cardiovascular magnetic resonance (CMR) imaging to perform wave intensity analysis non-invasively to shed light on VA coupling changes in BAV. The primary hypothesis was that BAV patients with COA exhibit unfavorable VA coupling, and the secondary hypothesis was that BAV patients with COA exhibit increased wave speed as a marker of reduced aortic distensibility despite successful surgical correction. Patients were retrospectively identified from a CMR database and divided into two groups: isolated BAV and BAV associated with repaired COA. Aortic and ventricular dimensions, global longitudinal strain (GLS), and ascending aortic flow data and area were collected and used to derive wave intensity from CMR data. The main variables for the analysis included all wave magnitudes (forward compression/expansion waves, FCW and FEW, respectively, and reflected backward compression wave, BCW) and wave speed. In the comparison of patients with isolated BAV and those with BAV associated with repaired COA (n = 25 in each group), no differences were observed in left ventricular ejection fraction, GLS, or ventricular volumes, whilst significant increases in FCW and FEW magnitude were noted in the BAV and repaired COA group. The FCW inversely correlated with age and aortic size. Whilst the BCW was not significantly different compared with that in patients with/without COA, its magnitude tends to increase with a lower COA index. Patients with repaired COA exhibited higher wave speed velocity. Aortic wave speed (inversely related to distensibility) was not significantly different between the two groups. In the absence of a significant restenosis, VA coupling in patients with BAV and COA is not negatively affected compared to patients with isolated BAV. A reduction in the magnitude of the early systolic FCW was observed in patients who were older and with larger aortic diameters.


Tephritid Integrative Taxonomy: Where We Are Now, with a Focus on the Resolution of Three Tropical Fruit Fly Species Complexes.
Accurate species delimitation underpins good taxonomy. Formalization of integrative taxonomy in the past decade has provided a framework for using multidisciplinary data to make species delimitation hypotheses more rigorous. We address the current state of integrative taxonomy by using as a case study an international project targeted at resolving three important tephritid species complexes: Bactrocera dorsalis complex, Anastrepha fraterculus complex, and Ceratitis FAR (C. fasciventris, C. anonae, C. rosa) complex. The integrative taxonomic approach has helped deliver significant advances in resolving these complexes: It has been used to identify some taxa as belonging to the same biological species as well as to confirm hidden cryptic diversity under a single taxonomic name. Nevertheless, the general application of integrative taxonomy has not been without issue, revealing challenges that must be considered when undertaking an integrative taxonomy project. Scrutiny of this international case study provides a unique opportunity to document lessons learned for the benefit of not only tephritid taxonomists, but also the wider taxonomic community.


Chi3L1 is a therapeutic target in bone metabolism and a potential clinical marker in patients with osteoporosis.
BMP2 is clinically used as an ectopic bone inducer and plays a significant role in bone development, formation, and diseases. Chitinase 3-like 1 protein (Chi3L1) is found in the skeletal system. However, Chi3L1-mediated bone metabolism and aging-related bone erosion via BMP2 signaling have not yet been demonstrated. Herein, Chi3L1 increased BMP2-induced osteoblast differentiation in mesenchymal precursor cells and human primary osteoblasts. Chi3L1KO(-/-) showed abnormal bone development, and primary osteoblasts isolated from Chi3L1KO(-/-) exhibited impaired osteoblast differentiation and maturation. Chi3L1 also potentiated BMP2 signaling and RUNX2 expression in primary osteoblasts. Chi3L1 interacted with BMPRIa, which increased the surface expression of BMPRIa and promoted BMP2 signaling to induce osteoblast differentiation. Chi3L1KO(-/-) mice showed bone formation reduced with a decrease in RUNX2 expression in calvarial defects. Chi3L1KO(-/-) mice exhibited aging-related osteoporotic bone loss with decreases in the levels of RUNX2 and OPG, while serum PYD level and osteoclast number increased. Chi3L1 increased OPG via non-canonical BMP2 signaling in osteoblasts, which suppressed osteoclastogenesis in BMMs. Furthermore, ROC analysis showed that serum Chi3L1 level clinically decreased in osteoporosis patients. Our findings demonstrate that Chi3L1 promotes bone formation, suppresses osteoclastogenesis, and prevents aging-related osteoporosis.


Structural studies on myosin II: communication between distant protein domains.
Understanding how chemical energy is converted into directed movement is a fundamental problem in biology. In higher organisms this is accomplished through the hydrolysis of ATP by three families of motor proteins: myosin, dynein and kinesin. The most abundant of these is myosin, which operates against actin and plays a central role in muscle contraction. As summarized here, great progress has been made towards understanding the molecular basis of movement through the determination of the three-dimensional structures of myosin and actin and through the establishment of systems for site-directed mutagenesis of this motor protein. It now appears that the generation of movement is coupled to ATP hydrolysis by a series of domain movements within myosin.


[Self-recognition need in the middle aged and elderly].
The purpose of this study was to investigate developmental changes in Self-Recognition Need (SRN; Kamise, 1992), measuring the need of middle aged and elder people. One hundred and ninety eight (198) men and women in their forties, and 199 in their sixties filled a questionnaire that measured SRN. Results showed that SRN declined with advancing years, and it was higher in women than in men. Targets of SRN also changed with advancing years. Men in their forties and women in their sixties directed their SRN toward their interpersonal and future selves, women in their forties toward their current and future selves, and men in their sixties toward their past selves.


A comparison between the effect of topical and systemic carbonic anhydrase inhibitors on aqueous humor secretion.
We have assessed the onset and duration of decreased intraocular pressure and aqueous humor flow contrasting systemic and topical administration of carbonic anhydrase inhibitors. The relationship between physiological effects and fractional activity of carbonic anhydrase isoenzymes in the eye was also investigated. Experiments were performed in normotensive New Zealand white rabbits. Intraocular pressure was determined manometrically or tonometrically and aqueous humor flow by sulfacetamide clearance. We studied methazolamide (25 mg kg-1), ethoxzolamide (4 mg kg-1), and MK-927 (2% in 0.5% hydroxyethylcellulose, topical, pH 4.8). There is an immediate reduction in intraocular pressure (1.2 and 1.8 mmHg by 2 min) and aqueous flow (33% and 40% by 5 min) following intravenous dosing with either methazolamide or ethoxzolamide. This correlates with rapid appearance of drug in the anterior uvea and very low fractional activity of ocular carbonic anhydrase isoenzymes II (cytosolic) and IV (membrane bound). Peak intraocular pressure reduction averaged 4.2 +/- 0.68 mmHg and 4.5 +/- 0.8 mmHg for methazolamide and ethoxzolamide at 60 and 45 min, respectively. Peak flow reduction was 38% for methazolamide and 40% for ethoxzolamide, at 5 min. Aqueous flow and intraocular pressure returned to baseline at 7 and 4 hr following methazolamide and ethoxzolamide, respectively. This corresponds to decay of drug from ocular tissues and significant increases in fractional activity of carbonic anhydrase isoenzymes. Topical MK-927 resulted in a 1.2 mmHg decrease in pressure by 5 min. This correlated with the early appearance of drug in the anterior uvea prior to its appearance in aqueous humor and very low fractional activity of carbonic anhydrase isoenzymes. Intraocular pressure decreased 3.6 +/- 0.35 mmHg at 1 hr and returned to baseline by 6 hr. Aqueous flow was reduced 12% by 5 min and 35% at 1 hr. The appearance of MK-927 in the anterior uvea prior to detection in aqueous suggests a significant non-corneal route of absorption following topical administration. Topical MK-927 results in a more gradual reduction in intraocular pressure and flow, although peak effects are not statistically different from systemic carbonic anhydrase inhibitors. The time of pressure return to baseline is also comparable to systemic carbonic anhydrase inhibitors. Because the relations between carbonic anhydrase II and carbonic anhydrase IV in the ciliary process are not yet clear and since the drugs have different affinities for the isozymes, the precise degree of fractional inhibition necessary for pharmacological effect is not certain, but based on drug concentration in the anterior uvea, may take 98% inhibition for full intraocular pressure reduction.


A regulatory mechanism involving TBP-1/Tat-Binding Protein 1 and Akt/PKB in the control of cell proliferation.
TBP-1 /Tat-Binding Protein 1 (also named Rpt-5, S6a or PSMC3) is a multifunctional protein, originally identified as a regulator of HIV-1-Tat mediated transcription. It is an AAA-ATPase component of the 19S regulative subunit of the proteasome and, as other members of this protein family, fulfils different cellular functions including proteolysis and transcriptional regulation. We and others reported that over expression of TBP-1 diminishes cell proliferation in different cellular contexts with mechanisms yet to be defined. Accordingly, we demonstrated that TBP-1 binds to and stabilizes the p14ARF oncosuppressor increasing its anti-oncogenic functions. However, TBP-1 restrains cell proliferation also in the absence of ARF, raising the question of what are the molecular pathways involved. Herein we demonstrate that stable knock-down of TBP-1 in human immortalized fibroblasts increases cell proliferation, migration and resistance to apoptosis induced by serum deprivation. We observe that TBP-1 silencing causes activation of the Akt/PKB kinase and that in turn TBP-1, itself, is a downstream target of Akt/PKB. Moreover, MDM2, a known Akt target, plays a major role in this regulation. Altogether, our data suggest the existence of a negative feedback loop involving Akt/PKB that might act as a sensor to modulate TBP-1 levels in proliferating cells.


Damage to UV-sensitive cells by short UV in photographic flashes.
Light emitted by electronic photographic flash units is shown to damage bacteria and human skin fibroblasts deficient in repair systems, with survival curves very similar to those produced by 254 nm short UV. The lesions induced by these flashes are as photorepairable by the photolyase enzyme as those induced by 254 nm UV and result in equivalent survival rates. Biological dosimetry performed with microorganisms highly sensitive to UV (Escherichia coli K12 AB2480, deficient in excision and recombinational-dependent repair systems and Bacillus subtilis UVSSP spores, deficient in excision and in a specific spore repair process) revealed that each 1 ms flash of light from the photographic unit used in this work contained the equivalent of 0.25 J m-2 of 254 nm UV, when measured at a distance of 7.0 cm. This dose of UV was found to be lethal to both repair-deficient E. coli bacteria and repair-deficient human skin fibroblasts obtained from xeroderma pigmentosum donors, as well as mutagenic in B/r wild-type and HCR-mutant bacteria.


Neuronal LRP4 regulates synapse formation in the developing CNS.
The low-density lipoprotein receptor-related protein 4 (LRP4) is essential in muscle fibers for the establishment of the neuromuscular junction. Here, we show that LRP4 is also expressed by embryonic cortical and hippocampal neurons, and that downregulation of LRP4 in these neurons causes a reduction in density of synapses and number of primary dendrites. Accordingly, overexpression of LRP4 in cultured neurons had the opposite effect inducing more but shorter primary dendrites with an increased number of spines. Transsynaptic tracing mediated by rabies virus revealed a reduced number of neurons presynaptic to the cortical neurons in which LRP4 was knocked down. Moreover, neuron-specific knockdown of LRP4 by in utero electroporation of LRP4 miRNA in vivo also resulted in neurons with fewer primary dendrites and a lower density of spines in the developing cortex and hippocampus. Collectively, our results demonstrate an essential and novel role of neuronal LRP4 in dendritic development and synaptogenesis in the CNS.


Diastereoisomers of cytolysins, a novel class of potent antibacterial peptides.
An amphipathic alpha-helical structure is considered to be a prerequisite for the lytic activity of most short linear cytolytic polypeptides that act on both mammalian cells and bacteria. This structure allows them also to exert diverse pathological and pharmacological effects, presumably by mimicking protein components that are involved in membrane-related events. In this study D-amino acid-incorporated analogues (diastereomers) of the cytolysin pardaxin, which is active against mammalian cells and bacteria, were synthesized and structurally and functionally characterized. We demonstrate that the diastereomers do not retain the alpha-helical structure, which in turn abolishes their cytotoxic effects on mammalian cells. However, they retain a high antibacterial activity, which is expressed in a complete lysis of the bacteria, as revealed by negative staining electron microscopy. The disruption of the alpha-helical structure should prevent the diastereomer analogues from permeating the bacterial wall by forming transmembrane pores but rather by dissolving the membrane as a detergent. These findings open the way for a new strategy in developing a novel class of highly potent antibacterial polypeptides for the treatment of infectious diseases, due to the increasing resistance of bacteria to the available antibacterial drugs.


An immunovirological study of central nervous system involvement during HIV-1 infection of chimpanzees.
Chimpanzees infected with human immunodeficiency virus type 1 (HIV-1) are used to model acquired immunodeficiency syndrome (AIDS). Since the central nervous system (CNS) is involved in AIDS, we performed an immunovirological study in 18 chimpanzees inoculated up to 87 months prior to the study (mean, 45 months) with HIV-1 and 8 uninfected controls. Serum and cerebrospinal fluid (CSF) IgG and albumin levels of infected chimpanzees never exceeded those of controls. The CSF/serum albumin ratio was elevated in 1 of 18 infected chimpanzees compared to controls; however, all animals had an elevated ratio indicating a more open blood-brain barrier relative to humans. The intrathecal IgG production index was elevated in only 1 of 18 infected chimpanzees compared to controls. Identical serum and CSF IgG bands were found by isoelectric focusing in 2 of 8 controls and in 1 of 18 infected chimpanzees. None of these bands reacted with recombinant HIV-1 p24gag or gp 120env. HIV-1 was isolated from the peripheral blood of 4 of 18 infected chimpanzees but never from the paired CSF samples. Anti-HIV-1 antibody was detected by a enzyme-linked immunosorbent assay in 18 of 18 paired serum and CSF samples and by Western blot in 18 of 18 serum and 13 of 18 CSF samples from infected chimpanzees without a difference in pattern. Polymerase chain reaction analysis on brain tissue of one animal was negative for HIV-1 sequences. Our results demonstrate that, unlike human infection, chimpanzees inoculated with HIV-1 show no evidence of isolatable virus in the CSF and no evidence of intrathecal anti-HIV-1 antibody synthesis up to several years after experimental infection. The lack of CNS involvement may contribute to the delay or suppression of clinical disease in infected chimpanzees.


Proapoptotic activity of bortezomib in gastrointestinal stromal tumor cells.
Gastrointestinal stromal tumors (GIST) are caused by activating mutations in the KIT or PDGFRA receptor tyrosine kinase genes. Although >85% of GIST patients treated with the small-molecule inhibitor imatinib mesylate (Gleevec) achieve disease stabilization, complete remissions are rare and a substantial proportion of patients develop resistance to imatinib over time. Upregulation of soluble, non-chromatin-bound histone H2AX has an important role in imatinib-induced apoptosis of GIST cells. Additionally, H2AX levels in untreated GIST are maintained at low levels by a pathway that involves KIT, phosphoinositide 3-kinase, and the ubiquitin-proteasome system. In this study, we asked whether bortezomib-mediated inhibition of the ubiquitin-proteasome machinery could lead to upregulation of histone H2AX and GIST cell death. We show that bortezomib rapidly triggers apoptosis in GIST cells through a combination of mechanisms involving H2AX upregulation and loss of KIT protein expression. Downregulation of KIT transcription was an underlying mechanism for bortezomib-mediated inhibition of KIT expression. In contrast, the nuclear factor-kappaB signaling pathway did not seem to play a major role in bortezomib-induced GIST cell death. Significantly, we found that bortezomib would induce apoptosis in two imatinib-resistant GIST cell lines as well as a short-term culture established from a primary imatinib-resistant GIST. Collectively, our results provide a rationale to test the efficacy of bortezomib in GIST patients with imatinib-sensitive or -resistant tumors.


Promoter analysis in the human SPRR gene family.
Protocols to study the regulation of a conserved multigene family (SPRR genes) during calcium-induced differentiation of cultured normal human keratinocytes (NHKs) are provided. Transfection of promoter-reporter (CAT or luciferase) constructs, combined with promoter truncation, can be used to study the expression of individual SPRR genes and to identify specific transcription factor binding sites. Interaction of regulatory factors with these control elements can be visualized and quantified by electro phoretic mobility shift analysis. Inclusion of specific antibodies in these experiments will identify the transcription factors involved in the observed mobility shift (supershift). A competitive electrophoretic mobility shift analysis, that is well suited to study the differential regulation of various SPRR members, is also described. These methods should be applicable to the study of other multigene families regulated during keratinocyte terminal differentiation.


Atypical Properties of a Thin Silver Layer Deposited on a Composite Textile Substrate.
Thin layers are widely used in electronics and protective coatings. They are also increasingly used in wearable electronics. A major challenge affecting the use of thin layers is their connection to flexible substrates, particularly textile products. This article describes the stability of the resistance of a silver layer with a thickness of 250 nm in a wide temperature range of 15-295 K. The aim was to determine the temperature dependence of the resistance of layers formed on a composite textile substrate compared with that of layers produced on an Al2O3 substrate. The results showed that the electrical parameters of the layer formed on the composite textile substrate changed in a manner atypical for metallic layers. This may have been due to the polyurethane base layer. The roughness and ability to deform under the influence of heat of the substrate can significantly affect the electrical parameters of a thin metal layer produced by the PVD coating process, which is important for the design of textronic applications.


ETV5 facilitates tumor progression in head-neck squamous cell carcinoma.
E26 transformation-specific (ETS) factors have emerged as key mediators underlying human tumorigenesis. Here, we sought to characterize the expression pattern, biological roles, and clinical significance of ETS Variant Transcription Factor 5 (ETV5) in head neck squamous cell carcinoma (HNSCC). ETV5 expression pattern in HNSCC was determined by bioinformatics interrogations and immunohistochemical staining in primary samples. The associations between its abundance with clinicopathological parameters, and patient survival were evaluated. Colony formation, CCK-8, flow cytometry, wound healing, and Transwell invasion assays, as well as xenograft models, were utilized to determine the phenotypic changes after ETV5 silencing in vitro and vivo. The potential binding of ETV5 in the Slug promoter was determined by ChIP-qPCR. ETV5 was significantly overexpressed in HNSCC samples. Its overexpression is significantly associated with aggressiveness features and reduced survival. ETV5 knockdown significantly inhibited cell proliferation, migration, invasion, and induced apoptosis in vitro, and impaired tumor growth in vivo. Moreover, ETV5-activated Slug transcription by binding its promoter region in HNSCC cells. Patients with ETV5high Slughigh had the worst survival across multiple HNSCC cohorts. Our findings reveal that ETV5 serves as a novel prognostic biomarker and putative oncogene for HNSCC progression likely by activating Slug transcription.


Results of surgery for portacaval shunt in a patient with situs inversus diagnosed and evaluated by per rectal portal scintigraphy.
A 70-yr-old man diagnosed with situs inversus was hospitalized because of hepatic encephalopathy. Per rectal portal scintigraphy showed portal hypertension and a region with high radionuclide activity in the right lateral region of the abdomen. In percutaneous transhepatic portograms, a giant portacaval shunt was seen in the region with high radionuclide activity. After the portacaval shunt was obstructed surgically, the hepatic encephalopathy disappeared. In per rectal portal scintigraphy done 2 wk after surgery, the pattern was normal and the region with high radionuclide activity had disappeared.


Induction of apoptosis in mesothelioma cells by antisurvivin oligonucleotides.
Malignant pleural mesothelioma is a rare and aggressive tumor characterized by rapid progression, late metastases, and poor prognosis. In this study, we investigated the expression of survivin, a member of the inhibitors of apoptosis protein gene family, in mesothelioma and an antisense oligonucleotide-based gene therapy for mesothelioma using survivin as a target. Initially, we documented the expression of survivin in human mesothelioma cell lines and fresh tissues using reverse transcription-PCR and Western blot analysis. Our results showed that survivin was overexpressed in 7 of 8 (87.5%) mesothelioma cell lines assayed and in all (12 of 12; 100%) freshly resected mesothelioma tissues analyzed. To investigate the use of survivin as a therapeutic target on mesothelioma, we carried out transfections with antisurvivin oligonucleotides to induce apoptosis in mesothelioma cell lines MS-1 and H28. Results from cellular transfection and subsequent analysis using the flow cytometry demonstrated that antisurvivin oligonucleotides induced significantly greater apoptosis rates in the survivin-positive mesothelioma cell line H28 (42.5%) as compared with the control oligonucleotides (16.2%; P < 0.001). The survivin-negative cell line LRK1A (survivin-/-) did not apoptose with antisense oligonucleotides. Furthermore, time course evaluation by Western blot analysis showed that survivin was inhibited by antisurvivin oligonucleotides within 12 h after transfection. Our results show, for the first time, that survivin, an inhibitors of apoptosis protein family gene member, is highly overexpressed in malignant pleural mesothelioma. Down-regulation of survivin by a targeted antisense oligonucleotide appears to be an effective gene therapy approach to the treatment of mesothelioma.


Virtual Laboratory and Imaging: an online simulation tool to enhance hospital disaster preparedness training experience.
Hospitals play a pivotal role as basic healthcare providers during mass casualty incidents (MCIs). Radiological studies and emergency laboratory test are of high importance for the management of hospital patients. However, it is known that during these events, they can generate significant bottlenecks. Appropriate request of such tests is of utmost importance to not generate delays in the patient flow. The aim of this paper is to describe a software designed to increase the realism of hospital-based MCI training through a realistic reproduction of radiology and laboratory departments. In this paper, we present a Virtual Laboratory and Imaging system that we designed with the goal of increasing the realism of full-scale mass casualty simulations. The system is able to dynamically manage the speed and load of virtual departments while collecting data on usage and load, and provide data useful for the after-event debriefing. We tested this system in two pilot simulations involving, respectively, 105 and 89 simulated casualties. The system, by measuring the number of requests and exams' turnaround time, enabled an objective measurement of the laboratory and radiology workload during simulated MCIs. It was possible to identify bottlenecks and consequently use these data for after-action debriefing. The tool not only increased the simulation realism by adding the radiology and laboratory departments but also provided valuable data that could be used for educational and organizational purposes.


Bridges, brokers and boundary spanners in collaborative networks: a systematic review.
Bridges, brokers and boundary spanners facilitate transactions and the flow of information between people or groups who either have no physical or cognitive access to one another, or alternatively, who have no basis on which to trust each other. The health care sector is a context that is rich in isolated clusters, such as silos and professional "tribes," in need of connectivity. It is a key challenge in health service management to understand, analyse and exploit the role of key agents who have the capacity to connect disparate groupings in larger systems. The empirical, peer reviewed, network theory literature on brokerage roles was reviewed for the years 1994 to 2011 following PRISMA guidelines. The 24 articles that made up the final literature set were from a wide range of settings and contexts not just healthcare. Methods of data collection, analysis, and the ways in which brokers were identified varied greatly. We found four main themes addressed in the literature: identifying brokers and brokerage opportunities, generation and integration of innovation, knowledge brokerage, and trust. The benefits as well as the costs of brokerage roles were examined. Collaborative networks by definition, seek to bring disparate groups together so that they can work effectively and synergistically together. Brokers can support the controlled transfer of specialised knowledge between groups, increase cooperation by liaising with people from both sides of the gap, and improve efficiency by introducing "good ideas" from one isolated setting into another.There are significant costs to brokerage. Densely linked networks are more efficient at diffusing information to all their members when compared to sparsely linked groups. This means that while a bridge across a structural hole allows information to reach actors that were previously isolated, it is not the most efficient way to transfer information. Brokers who become the holders of, or the gatekeepers to, specialised knowledge or resources can become overwhelmed by the role and so need support in order to function optimally.