Shared-Use Decisions Among Administrators of Physical Activity Facilities in Pasadena, TX.
Background and Purpose. The American Heart Association recommends community-based research on shared use of physical activity (PA) spaces. Pasadena, a community in southeast Houston, Texas with lower socioeconomic status and racial/ethnic diversity, was the setting for our study. Efforts to increase access to PA in Pasadena include building the evidence on PA resources. The purpose of this research was to utilize survey data in a community setting to inform and target efforts around sharing PA spaces. Method. An online survey was administered to K-12 school (n = 25) and park (n = 30) administrators, church leaders (n = 10), community organizations (n = 2), a health care center, and a local business. Results. Park facilities in Pasadena shared by agreement with two high schools and three baseball/softball leagues were ballfields, tennis courts, and jogging paths. No park facilities were shared with faith-based organizations. Four parks communicated daily, and five schools communicated with parks quarterly about providing opportunities for PA. Key facilitators to sharing facilities were building relationships and collaboration, service to the community, and improving health. Key barriers were cost, maintenance, staffing, and prioritizing use with limited time, facilities, and resources. Conclusions. This study is the first of its kind to address shared use at the community level and suggests opportunities to improve communication and partnerships between parks, schools, and churches. This research will inform ongoing efforts to promote access by identifying barriers and motivators among stakeholder groups to help facilitate shared use agreements.


Isolation and Characterization of Live Yeast Cells from Ancient Vessels as a Tool in Bio-Archaeology.
Ancient fermented food has been studied based on recipes, residue analysis, and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach based on our hypothesis that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and their descendants could be isolated and studied today. We developed a pipeline of yeast isolation from clay vessels and screened for yeast cells in beverage-related and non-beverage-related ancient vessels and sediments from several archaeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. The findings that genotypically the isolated yeasts are similar to those found in traditional African beverages and phenotypically they grow similar to modern beer-producing yeast strongly suggest that they are descendants of the original fermenting yeast. These results demonstrate that modern microorganisms can serve as a new tool in bio-archaeology research.IMPORTANCE So far, most of the study of ancient organisms has been based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms-yeast in this case-from ancient pottery vessels used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains that participated in the fermentation process and were absorbed into the clay matrix of the pottery vessels. Moreover, we characterized the isolated yeast strains, their genomes, and the beer they produced. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio- and experimental archaeology that aim to reconstruct ancient artifacts and products.


Coronin-1C and RCC2 guide mesenchymal migration by trafficking Rac1 and controlling GEF exposure.
Sustained forward migration through a fibrillar extracellular matrix requires localization of protrusive signals. Contact with fibronectin at the tip of a cell protrusion activates Rac1, and for linear migration it is necessary to dampen Rac1 activity in off-axial positions and redistribute Rac1 from non-protrusive membrane to the leading edge. Here, we identify interactions between coronin-1C (Coro1C), RCC2 and Rac1 that focus active Rac1 to a single protrusion. Coro1C mediates release of inactive Rac1 from non-protrusive membrane and is necessary for Rac1 redistribution to a protrusive tip and fibronectin-dependent Rac1 activation. The second component, RCC2, attenuates Rac1 activation outside the protrusive tip by binding to the Rac1 switch regions and competitively inhibiting GEF action, thus preventing off-axial protrusion. Depletion of Coro1C or RCC2 by RNA interference causes loss of cell polarity that results in shunting migration in 1D or 3D culture systems. Furthermore, morpholinos against Coro1C or RCC2, or mutation of any of the binding sites in the Rac1-RCC2-Coro1C complex delays the arrival of neural crest derivatives at the correct location in developing zebrafish, demonstrating the crucial role in migration guidance in vivo.


Targeted Delivery of Neutralizing Anti-C5 Antibody to Renal Endothelium Prevents Complement-Dependent Tissue Damage.
Complement activation is largely implicated in the pathogenesis of several clinical conditions and its therapeutic neutralization has proven effective in preventing tissue and organ damage. A problem that still needs to be solved in the therapeutic control of complement-mediated diseases is how to avoid side effects associated with chronic neutralization of the complement system, in particular, the increased risk of infections. We addressed this issue developing a strategy based on the preferential delivery of a C5 complement inhibitor to the organ involved in the pathologic process. To this end, we generated Ergidina, a neutralizing recombinant anti-C5 human antibody coupled with a cyclic-RGD peptide, with a distinctive homing property for ischemic endothelial cells and effective in controlling tissue damage in a rat model of renal ischemia/reperfusion injury (IRI). As a result of its preferential localization on renal endothelium, the molecule induced complete inhibition of complement activation at tissue level, and local protection from complement-mediated tissue damage without affecting circulating C5. The ex vivo binding of Ergidina to surgically removed kidney exposed to cold ischemia supports its therapeutic use to prevent posttransplant IRI leading to delay of graft function. Moreover, the finding that the ex vivo binding of Ergidina was not restricted to the kidney, but was also seen on ischemic heart, suggests that this RGD-targeted anti-C5 antibody may represent a useful tool to treat organs prior to transplantation. Based on this evidence, we propose preliminary data showing that Ergidina is a novel targeted drug to prevent complement activation on the endothelium of ischemic kidney.


The peplomer protein E2 of coronavirus JHM as a determinant of neurovirulence: definition of critical epitopes by variant analysis.
We selected murine coronavirus JHM variants specifically changed in defined antigenic sites of the peplomer protein E2. Variants were isolated from the supernatants of monoclonal antibody hybridoma cell cultures which continued to secrete neutralizing antibodies after being infected with JHM. Comparative antigenic analysis and biological tests were performed in order to refine an operational epitope map and to characterize functional domains important for pathogenicity. The reaction patterns (neutralization, inhibition of cell fusion, immunofluorescence and binding in ELISA) between the variant viruses and the panel of monoclonal antibodies were very similar. Four groups of variants were characterized each of which revealed distinct changes affecting one defined antigenic site. These observations indicated that at least four independently mutable antigenic sites were associated with domains involved in cell fusion, neutralization and pathogenicity (E2-Aa, -Ab, -Ba and -Bb). JHM variants with alterations in the E2-Aa, -Ab or -Bb sites were similar to wild-type virus. These variants caused acute hepatitis and encephalomyelitis in mice. In contrast, JHM variants with changes in site E2-Ba had a strong propensity to induce chronic disease accompanied by demyelination persisting for several months.


A CagA-independent cluster of antigens related to the risk of noncardia gastric cancer: associations between Helicobacter pylori antibodies and gastric adenocarcinoma explored by multiplex serology.
Because of the differences in bacterial epitopes and host characteristics, infections with Helicobacter pylori (H. pylori) induce different immune responses. We explored the possibility that certain antibody response patterns are more closely linked to gastric adenocarcinoma (GAC) than others. In a Swedish population-based case-control study, serum samples were obtained from 268 cases and 222 controls, aged 40-79 years and frequency-matched according to age and sex. We measured antibodies against 17 H. pylori proteins using multiplex serology. Associations were estimated with multivariably adjusted logistic regression models, using odds ratio (OR) with 95% confidence interval (CI) as measures of relative risk. Associations were essentially confined to non-cardia GAC but did not differ significantly between intestinal and diffuse subtypes. Point estimates for all antibodies were above unity, 15 significant with top three being CagA (OR = 9.2), GroEL (6.6), HyuA (3.6). ORs were substantially attenuated in individuals with chronic atrophic gastritis. Principal component analysis identified two significant factors: a CagA-dominant factor (antibodies against CagA, VacA and Omp as prominent markers), and a non-CagA factor (antibodies against NapA and Catalase as prominent markers). Both factors showed dose-dependent associations with non-cardia GAC risk (CagA-dominant factor, highest vs. lowest quartiles, OR = 16.2 [95% CI 4.8-54.9]; non-CagA factor OR = 5.3 [95% CI 2.1-13.3]). Overall, our results confirm that serum antibodies against different H. pylori proteins are associated with the presence of non-cardia GAC. Although strongest association is detected by antibodies against CagA and covarying proteins, a pattern of antibodies unrelated to CagA is also significantly linked to the risk of non-cardia GAC.


Stroke during pregnancy: no increased risk of preterm delivery and low birth weight, a nationwide case-controlled study.
Population-based data regarding pregnancy outcomes for patients who experienced stroke during pregnancy is sparse and mainly from Western populations. The authors investigate the association between strokes and pregnancy outcomes in Chinese women through a nationwide case-controlled study. A total of 161 women having live singleton births between 2001 and 2003 who had strokes during pregnancy were included in the study group. The authors randomly selected 1288 mothers matched in age and year of delivery for the comparison group. Conditional logistic regressions were performed to examine the odds of low birth weight (LBW), preterm and small-for-gestational-age (SGA) babies between these two groups, after adjusting for the characteristics of mother and infant and comorbid medical disorders. The authors found that the mothers with stroke during pregnancy were more likely to have lower monthly family incomes (p=0.031) and to have gestational hypertension (p=0.004), anaemia (p<0.001) and pre-eclampsia/eclampsia (p=0.039) than the comparison group. Results showed no significant differences in the prevalence of preterm births (5.6% vs 7.1%, p=0.466), LBW (5.6% vs 7.1%, p=0.486) and SGA infants (14.3% vs 16.9%, p=0.396) between women who had strokes during pregnancy and mothers in the comparison group. After adjusting for other potential confounders, conditional logistic regression analyses showed no significant differences in the odds of having preterm, LBW babies and SGA infants between two groups. The authors found that there is no statistically significant difference in the pregnancy outcomes between Chinese women who had strokes during pregnancy and those who did not.


Airway Changes after Cleft Orthognathic Surgery Evaluated by Three-Dimensional Computed Tomography and Overnight Polysomnographic Study.
Cleft lip and palate is the most common congenital craniofacial anomaly. Up to 60% of these patients will benefit from cleft orthognathic surgery, which consists primarily of maxillary advancement and mandibular setback to address midface retrusion and relative mandibular protrusion, respectively. It is believed that maxillary advancement can enlarge the airway whilst mandibular setback can reduce the airway, but this has not previously been quantified for cleft patients undergoing orthognathic surgery. This unique longitudinal prospective study of 18 patients was conducted between April 2013 and July 2016. No significant changes occurred by six months postoperatively in body mass index, apnoea-hypopnoea index or lowest oxygen saturation (LSAT). There was a mean increase of 0.73 cm3 in velopharyngeal volume, a mean decrease of 0.79 cm3 in oropharyngeal volume, an improvement in snoring index, and no statistically significant change in hypopharyngeal volume. In conclusion, cleft orthognathic surgery that produced anterior advancement of the maxilla, setback of the mandible and clockwise rotation of the maxillo-mandibular complex resulted in increased velopharyngeal, decreased oropharyngeal and unchanged hypopharyngeal airways, and improved snoring, but did not significantly alter objective sleep-related breathing function.


Improvement and quantitative performance estimation of the back support muscle suit.
We have been developing the wearable muscle suit for direct and physical motion supports. The use of the McKibben artificial muscle has opened the way to the introduction of "muscle suits" compact, lightweight, reliable, wearable "assist-bots" enabling manual worker to lift and carry weights. Since back pain is the most serious problem for manual worker, improvement of the back support muscle suit under the feasibility study and quantitative estimation are shown in this paper. The structure of the upper body frame, the method to attach to the body, and the axes addition were explained as for the improvement. In the experiments, we investigated quantitative performance results and efficiency of the back support muscle suit in terms of vertical lifting of heavy weights by employing integral electromyography (IEMG). The results indicated that the values of IEMG were reduced by about 40% by using the muscle suit.


An evaluation of a plastic strip thermometer.
The Clinitemp Fever Detector (a registered trademark for the Clinitemp Co, Indianapolis) is a plastic strip thermometer designed to be placed on the forehead and to detect body temperature by changing color. In a sample of 613 patients, we evaluated the ability of the Clinitemp to detect fevers detected by traditional mercury/glass thermometers. When a Clinitemp reading of 37.8 degrees C (100 degrees F) was used to demark fever, the Clinitemp detected 34% of the clinically significant fevers detected by the mercury/glass thermometers. The sensitivity of the Clinitemp rose to .81 when a Clinitemp reading of 37.2 degrees C (99 degrees F) was used to demark fever. The accuracy of the Clinitemp was affected by variations in ambient air temperature. A reassessment of previously published evaluations of plastic strip thermometers suggest that our results are consistent with other experimental findings. We conclude that the Clinitemp is unacceptable as a substitute for the mercury/glass thermometer.


Temporal Trends Of Pharmacologic Therapies For Patients With Chronic Obstructive Pulmonary Disease In Alberta, Canada.
To describe the trends in pharmacologic treatment for patients newly diagnosed with chronic obstructive pulmonary disease (COPD) in Alberta, Canada. We linked Alberta health databases to identify patients aged ≥35 years with incident COPD between April 2010 and March 2017. Incident cases were defined as those who did not have a hospitalization or outpatient visit with COPD in the previous 2 years. Patients were categorized into two groups: 1) incident cases at a hospital and 2) incident cases at an outpatient clinic, and both were followed until death or being censored by 31 March 2018. Utilization of COPD medication for 30 days following incident event and adherence in maintenance therapy over time were reported. The study included 33,169 patients with incident COPD (hospital: 9,089; outpatient: 24,080). In 18,666 (56.3%) patients starting medication within 30 days of the incident event (2010: 52.7%; 2016: 56.6%; p=0.002), SABA (60.5%) and LABA/ICS (41.6%) were most commonly used. ICS (without LABA) was used in 14.2% and was used as monotherapy in 4.5% of patients. The proportion of patients who initiated any ICS was similar (hospital: 56.7%; outpatient: 55.7%; p=0.194) and decreased in both settings over time (p<0.001). Drug adherence during the first year after the incident event was 54.3%, higher among hospital patients (66.5% vs 48.9%; p<0.001), and improved over time (2010: 53.4%; 2016: 57.4%; p<0.001). The initiation of and adherence to pharmacologic therapy for patients with COPD is low but improves over time. While SABA and LABA/ICS are most commonly used, ICS utilization decreases over time.


Metabolic response to Parkinson's disease recapitulated by the haploinsufficient diploid yeast cells hemizygous for the adrenodoxin reductase gene.
Adrenodoxin reductase, a widely conserved mitochondrial P450 protein, catalyses essential steps in steroid hormone biosynthesis and is highly expressed in the adrenal cortex. The yeast adrenodoxin reductase homolog, Arh1p, is involved in cytoplasmic and mitochondrial iron homeostasis and is required for activity of enzymes containing an Fe-S cluster. In this paper, we investigated the response of yeast to the loss of a single copy of ARH1, an oxidoreductase of the mitochondrial inner membrane, which is among the few mitochondrial proteins that is essential for viability in yeast. The phenotypic, transcriptional, proteomic, and metabolic landscape indicated that Saccharomyces cerevisiae successfully adapted to this loss, displaying an apparently dosage-insensitive cellular response. However, a considered investigation of transcriptional regulation in ARH1-impaired yeast highlighted that a significant hierarchical reorganisation occurred, involving the iron assimilation and tyrosine biosynthetic processes. The interconnected roles of the iron and tyrosine pathways, coupled with oxidative processes, are of interest beyond yeast since they are involved in dopaminergic neurodegeneration associated with Parkinson's disease. The identification of similar responses in yeast, albeit preliminary, suggests that this simple eukaryote could have potential as a model system for investigating the regulatory mechanisms leading to the initiation and progression of early disease responses in humans.


p53-dependent acinar cell apoptosis triggers epithelial proliferation in duct-ligated murine pancreas.
The mechanisms linking acinar cell apoptosis and ductal epithelial proliferation remain unknown. To determine the relationship between these events, pancreatic duct ligation (PDL) was performed on p53(+/+) and p53(-/-) mice. In mice bearing a wild-type p53 allele, PDL resulted in upregulation of p53 protein in both acinar cells and proliferating duct-like epithelium. In contrast, upregulation of Bcl-2 occurred only in duct-like epithelium. Both p21(WAF1/CIP1) and Bax were also upregulated in duct-ligated lobes. After PDL in p53(+/+) mice, acinar cells underwent widespread apoptosis, while duct-like epithelium underwent proliferative expansion. In the absence of p53, upregulation of p53 target genes and acinar cell apoptosis did not occur. The absence of acinar cell apoptosis in p53(-/-) mice also eliminated the proliferative response to duct ligation. These data demonstrate that PDL-induced acinar cell apoptosis is a p53-dependent event and suggest a direct link between acinar cell apoptosis and proliferation of duct-like epithelium in duct-ligated pancreas.


Role of Local Evidence in Transferring Evidence-Based Interventions to Low- and Middle-Income Country Settings: Application to Global Cancer Prevention and Control.
Although the global burden of cancer falls increasingly on low- and middle-income countries (LMICs), much of the evidence for cancer prevention and control comes from high-income countries and may not be directly applicable to LMIC settings. In this paper, we focus on the following question: When the majority of the evidence supporting an evidence-based intervention or implementation strategy comes from high-income countries, what local, contextual evidence is needed when transferring and adapting an intervention or strategy to a specific LMIC setting? We draw on an existing framework (the Population, Intervention, Environment, Transfer-T process model) for assessing transferability of interventions between distinct settings and apply the model to two case studies as learning examples involving implementation of tobacco use treatment guidelines and self sampling for human papillomavirus DNA in cervical cancer screening. These two case studies illustrate how researchers, policymakers, practitioners, and consumers may approach the need for local evidence from different perspectives and with different priorities. As uses and expectations around local evidence may be different for different groups, aligning these priorities through multistakeholder engagement in which all parties participate in defining the questions and cocreating the solutions is critical, along with promoting standardized reporting of contextual factors. Local, contextual evidence can be important for both researchers and practitioners, and its absence may hinder translation of research and implementation efforts across different settings. However, it is essential for researchers, practitioners, and other stakeholders to be able to clearly articulate the type of data needed and why it is important. In particular, where resources are limited, evidence generation should be prioritized to address real needs and gaps in knowledge.


Synthetic multi-contrast late gadolinium enhancement imaging using post-contrast magnetic resonance fingerprinting.
Late gadolinium enhancement (LGE) MRI is the non-invasive reference standard for identifying myocardial scar and fibrosis but has limitations, including difficulty delineating subendocardial scar and operator dependence on image quality. The purpose of this work is to assess the feasibility of generating multi-contrast synthetic LGE images from post-contrast T1 and T2 maps acquired using magnetic resonance fingerprinting (MRF). Fifteen consecutive patients with a history of prior ischemic cardiomyopathy (12 men; mean age 63 ± 13 years) were prospectively scanned at 1.5 T between Oct 2020 and May 2021 using conventional LGE and MRF after injection of gadolinium contrast. Three classes of synthetic LGE images were derived from MRF post-contrast T1 and T2 maps: bright-blood phase-sensitive inversion recovery (PSIR), black- and gray-blood T2 -prepared PSIR (T2 -PSIR), and a novel "tissue-optimized" image to enhance differentiation among scar, viable myocardium, and blood. Image quality was assessed on a 1-5 Likert scale by two cardiologists, and contrast was quantified as the mean absolute difference (MAD) in pixel intensities between two tissues, with different methods compared using Kruskal-Wallis with Bonferroni post hoc tests. Per-patient and per-segment scar detection rates were evaluated using conventional LGE images as reference. Image quality scores were highest for synthetic PSIR (4.0) and reference images (3.8), followed by synthetic tissue-optimized (3.3), gray-blood T2 -PSIR (3.0), and black-blood T2 -PSIR (2.6). Among synthetic images, PSIR yielded the highest myocardium/scar contrast (MAD = 0.42) but the lowest blood/scar contrast (MAD = 0.05), and vice versa for T2 -PSIR, while tissue-optimized images achieved a balance among all tissues (myocardium/scar MAD = 0.16, blood/scar MAD = 0.26, myocardium/blood MAD = 0.10). Based on reference mid-ventricular LGE scans, 13/15 patients had myocardial scar. The per-patient sensitivity/accuracy for synthetic images were the following: PSIR, 85/87%; black-blood T2 -PSIR, 62/53%; gray-blood T2 -PSIR, 100/93%; tissue optimized, 100/93%. Synthetic multi-contrast LGE images can be generated from post-contrast MRF data without additional scan time, with initial feasibility shown in ischemic cardiomyopathy patients.


Dissecting the Impact of the Gut Microbiome on Cancer Immunotherapy.
The gut microbiome has emerged as a key regulator of response to cancer immunotherapy. However, there is a gap in our understanding of the underlying mechanisms by which the microbiome influences immunotherapy. To this end, we developed a mathematical model based on i) gut microbiome data derived from preclinical studies on melanomas after fecal microbiota transplant, ii) mechanistic modeling of antitumor immune response, and iii) robust association analysis of murine and human microbiome profiles with model-predicted immune profiles. Using our model, we could distill the complexity of these murine and human studies on microbiome modulation in terms of just two model parameters: the activation and killing rate constants of immune cells. We further investigated associations between specific bacterial taxonomies and antitumor immunity and immunotherapy efficacy. This model can guide the design of studies to refine and validate mechanistic links between the microbiome and immune system.


Prehospital care of tsunami victims in Thailand: description and analysis.
On 26 December 2004 at 09:00 h, an earthquake of 9.0 magnitude (Richter scale) struck the area off of the western coast of northern Sumatra, Indonesia, triggering a Tsunami. As of 25 January 2005, 5,388 fatalities were confirmed, 3,120 people were reported missing, and 8,457 people were wounded in Thailand alone. Little information is available in the medical literature regarding the response and restructuring of the prehospital healthcare system in dealing with major natural disasters. The objective of the study was to analyze the prehospital medical response to the Tsunami in Thailand, and to identify possible ways of improving future preparedness and response. The Israeli Defense Forces (IDF) Home Front Command Medical Department sent a research delegation to study the response of the Thai medical system to the 2004 earthquake and Tsunami disaster. The delegation met with Thai healthcare and military personnel, who provided medical care for and evacuated the Tsunami victims. The research instruments included questionnaires (open and closed questions), interviews, and a review of debriefing session reports held in the days following the Tsunami. Beginning the day after the event, primary health care in the affected provinces was expanded and extended. This included: (1) strengthening existing primary care facilities with personnel and equipment; (2) enhancing communication and transportation capabilities; (3) erecting healthcare facilities in newly constructed evacuation centers; (4) deploying mobile, medical teams to make house calls to flood refugees in affected areas; and (5) deploying ambulance crews to the affected areas to search for survivors and provide primary care triage and transportation. The restructuring of the prehospital healthcare system was crucial for optimal management of the healthcare needs of Tsunami victims and for the reduction of the patient loads on secondary medical facilities. The disaster plan of a national healthcare system should include special consideration for the restructuring and reinforcement prehospital system.


Understanding temporal changes and seasonal variations in glycemic trends using wearable data.
Seasonal variations in glycemic trends remain largely unstudied despite the growing prevalence of diabetes. To address this gap, our objective is to investigate temporal changes in glycemic trends by analyzing intensively sampled blood glucose data from 137 patients (ages 2 to 76, primarily type 1 diabetes) over the course of 9 months to 4.5 years. From over 91,000 days of continuous glucose monitor data, we found that glycemic control decreases significantly around the holidays, with the largest decline observed on New Year's Day among the patients with already poor glycemic control (i.e., <55% time in the target range). We also observed seasonal variations in glycemic trends, with patients having worse glycemic control in the months of November to February (i.e., mid-fall and winter, in the United States), and better control in the months of April to August (i.e., mid-spring and summer). These insights are critical to inform targeted interventions that can improve diabetes outcomes.


Concussion Evaluation and Management: An Osteopathic Perspective.
Concussions have been increasingly reported over the past decade, but the reported incidence likely minimizes the actual numbers of people affected. Associated symptoms include emotional, somatic, and cognitive complaints, which may be prolonged in patients with certain risk factors. Neurologic examination is necessary to exclude upper motor neuron lesions and thus the need for brain imaging. Cervical conditions are often found concurrently with head injury and displays a similar presentation to concussions. Therefore, determining symptom origin can be problematic. Neuropsychological, oculomotor, and balance evaluations expose specific deficits that can be successfully managed with rehabilitation. Osteopathic assessment of the cranium, spine, sacrum, and thorax for somatic dysfunctions allows for prudent interventions. Patients involved in sports may begin an established graduated return-to-play protocol once cleared by their physician. Concurrently, a parallel return-to-learn program, with applicable academic accommodations, is recommended.


Acute intrinsic renal failure in cats: 32 cases (1997-2004).
To determine patient demographics, clinicopathologic findings, and outcome associated with naturally acquired acute intrinsic renal failure (ARF) in cats. Retrospective case series. 32 cats with ARF. Cats were considered to have ARF if they had acute onset of clinical signs (< 7 days), serum creatinine concentration > 2.5 mg/dL (reference range, 0.8 to 2.3 mg/dL) and BUN > 35 mg/dL (reference range, 15 to 34 mg/dL) in conjunction with urine specific gravity < 1.025 or with anuria or increasing serum creatinine concentration despite fluid therapy and normal hydration status, and no signs of chronic renal disease. Cases were excluded if cats had renal calculi or renal neoplasia. Causes of ARF included nephrotoxins (n = 18 cats), ischemia (4), and other causes (10). Eighteen cats were oliguric. For each unit (mEq/L) increase in initial potassium concentration, there was a 57% decrease in chance of survival. Low serum albumin or bicarbonate concentration at initial diagnosis was a negative prognostic indicator for survival. Initial concentrations of BUN, serum creatinine, and other variables were not prognostic. Seventeen (53%) cats survived, of which 8 cats had resolution of azotemia and 9 cats were discharged from the hospital with persistent azotemia. Results suggested that survival rates of cats with ARF were similar to survival rates in dogs and that residual renal damage persisted in approximately half of cats surviving the initial hospitalization.


Well-differentiated endocrine cell carcinoma of ileum treated by laparoscopy-assisted surgery--a case report.
A 72-year-old woman presented at our hospital with a 1-year history of intermittent right lower abdominal pain. Colonoscopic examination revealed a submucosal tumor with a pitted surface in the terminal ileum. Histopathological diagnosis of the carcinoid tumor was made following biopsy. Blood serotonin and urine 5-hydroxy-indoleacetic acid levels were normal, and carcinoid syndrome was not detected. Enhanced abdominal computed tomography scan and 18F-fluorodeoxyglucose positron emission tomography failed to detect multiple lesions, lymph node swelling or distant metastasis. Laparoscopy-assisted ileocecal resection with lymph node dissection was performed. The resected specimen showed a submucosal tumor with a pitted surface 11 x 11 mm in size, located at the terminal ileum. Histopathological examination revealed a well-differentiated endocrine cell carcinoma with an invasion depth to the muscularis propria. Immunohistochemical analysis showed the tumor cells to be chromogranin A and CD56-positive. The patient had no sign of recurrence for 16 months.


Differentiation of postnatal cerebellar glial progenitors is controlled by Bmi1 through BMP pathway inhibition.
Bmi1 is a polycomb group (Pc-G) protein involved in heritable gene repression, maintenance of cell identity, and proliferation. During the development of the central nervous system, Bmi1 is crucial for self-renewal of neural stem cells and for proliferation of neuronal (granule cell) progenitors of the cerebellum. Here, we use loss of function mouse models and in vitro assays--granule cell cultures and glial-neuronal co-cultures--to show that Bmi1 plays a crucial role in specification of glial progenitors during postnatal cerebellar development. Moreover, we demonstrate in in vitro assays that Bmi1 exerts this novel function through repression of BMP pathway and that this is independent of its known role in mediating the cellular response to Shh signaling. Thus modulation of Bmi1 expression in glial progenitors may represent a key event in determining the differentiation potential of these cells.


The bipedicled latissimus dorsi myocutaneous free flap: clinical experience with 53 patients.
The Latissimus dorsi musculocutaneous flap is a valuable workhorse of the microsurgeon, especially in closing large body defects. One of the pitfalls in harvesting the flap, is particularly in its inferior aspect which may be unreliable. Here we report a series of 53 patients who were undergone bipedicled free latissimus dorsi musculocutaneous free flaps for extensive tissue defects. The age of patients were between 5 and 64 and all of them were males. The wound sizes in these patients ranged between 31-35 x 10-12 cm and flap dimensions were between 38-48 x 6-8 cm. Perforator branches of the 10th intercostal vessels were dissected and supercharged to the flaps to reduce the risk of ischemia of the inferior cutaneous extensions. The secondary pedicles were anastomosed to recipient vessels other than the primary pedicles. Recipient areas were consisted of lower extremities. Four patients suffered of early arterial failure in the major pedicle and all revisions were successfully attempted. Neither sign of venous congestion nor arterial insufficiency were observed at the inferior cutaneous extensions of the flaps, and all defects were reconstructed successfully. All donor sites were primarily closed, only two patients suffered from a minor area of superficial epidermal loss at the donor site, without suffering any adjunct complications. In conclusion coverage of large defects can be safely performed with extending the skin paddle of latissimus dorsi flap as a bipedicled free flap.


Phosphorus toxicity disrupts Rubisco activation and reactive oxygen species defence systems by phytic acid accumulation in leaves.
Phosphorus (P) is an essential mineral nutrient for plants. Nevertheless, excessive P accumulation in leaf mesophyll cells causes necrotic symptoms in land plants; this phenomenon is termed P toxicity. However, the detailed mechanisms underlying P toxicity in plants have not yet been elucidated. This study aimed to investigate the molecular mechanism of P toxicity in rice. We found that under excessive inorganic P (Pi) application, Rubisco activation decreased and photosynthesis was inhibited, leading to lipid peroxidation. Although the defence systems against reactive oxygen species accumulation were activated under excessive Pi application conditions, the Cu/Zn-type superoxide dismutase activities were inhibited. A metabolic analysis revealed that excessive Pi application led to an increase in the cytosolic sugar phosphate concentration and the activation of phytic acid synthesis. These conditions induced mRNA expression of genes that are activated under metal-deficient conditions, although metals did accumulate. These results suggest that P toxicity is triggered by the attenuation of both photosynthesis and metal availability within cells mediated by phytic acid accumulation. Here, we discuss the whole phenomenon of P toxicity, beginning from the accumulation of Pi within cells to death in land plants.


[Analysis of displacement currents in lipid bilayer membranes].
The time course of a charge transfer through solvent-containing lipid bilayer membranes was measured in the range of 5 microseconds and 100 s. It is shown that a displacement charge value is proportional to the area of a torus surrounding the lipid bilayer membrane, but not to the black membrane area.


Spatially Heterogeneous Surface Water Diffusivity around Structured Protein Surfaces at Equilibrium.
Hydration water on the surface of a protein is thought to mediate the thermodynamics of protein-ligand interactions. For hydration water to play a role beyond modulating global protein solubility or stability, the thermodynamic properties of hydration water must reflect on the properties of the heterogeneous protein surface and thus spatially vary over the protein surface. A potent read-out of local variations in thermodynamic properties of hydration water is its equilibrium dynamics spanning picosecond to nanosecond time scales. In this study, we employ Overhauser dynamic nuclear polarization (ODNP) to probe the equilibrium hydration water dynamics at select sites on the surface of Chemotaxis Y (CheY) in dilute solution. ODNP reports on site-specific hydration water dynamics within 5-10 Å of a label tethered to the biomolecular surface on two separate time scales of motion, corresponding to diffusive water (DW) and protein-water coupled motions, referred to as bound water (BW). We find DW dynamics to be highly heterogeneous across the surface of CheY. We identify a significant correlation between DW dynamics and the local hydropathy of the CheY protein surface, empirically determined by molecular dynamics (MD) simulations, and find the more hydrophobic sites to be hydrated with slower diffusing water. Furthermore, we compare the hydration water dynamics on different polypeptides and liposome surfaces and find the DW dynamics on globular proteins to be significantly more heterogeneous than on intrinsically disordered proteins (IDPs), peptides, and liposomes. The heterogeneity in the hydration water dynamics suggests that structured proteins have the capacity to encode information into the surrounding hydration shell.


A proteomic strategy for global analysis of plant protein complexes.
Global analyses of protein complex assembly, composition, and location are needed to fully understand how cells coordinate diverse metabolic, mechanical, and developmental activities. The most common methods for proteome-wide analysis of protein complexes rely on affinity purification-mass spectrometry or yeast two-hybrid approaches. These methods are time consuming and are not suitable for many plant species that are refractory to transformation or genome-wide cloning of open reading frames. Here, we describe the proof of concept for a method allowing simultaneous global analysis of endogenous protein complexes that begins with intact leaves and combines chromatographic separation of extracts from subcellular fractions with quantitative label-free protein abundance profiling by liquid chromatography-coupled mass spectrometry. Applying this approach to the crude cytosolic fraction of Arabidopsis thaliana leaves using size exclusion chromatography, we identified hundreds of cytosolic proteins that appeared to exist as components of stable protein complexes. The reliability of the method was validated by protein immunoblot analysis and comparisons with published size exclusion chromatography data and the masses of known complexes. The method can be implemented with appropriate instrumentation, is applicable to any biological system, and has the potential to be further developed to characterize the composition of protein complexes and measure the dynamics of protein complex localization and assembly under different conditions.


Chronic disease knowledge and its determinants among chronically ill adults in rural areas of Shanxi Province in China: a cross-sectional study.
Chronic disease knowledge is an important prerequisite for an individual to implement behavioural changes towards the prevention and control of chronic diseases (CDs). Limited information is available about the relationship between different levels of health services and CD knowledge among rural residents with CDs. This research explores the distribution characteristics of CD knowledge and its determinants among chronically ill adults in rural China according to the aspects of patients and health service providers. A cross-sectional study was undertaken to estimate distribution characteristics of CD knowledge and collect data of socio-demographic characteristics, healthcare institutions attendances, duration of illness, and family history of CDs. Participants were 1060 rural adults with hypertension or type II diabetes. Correct responses to 12 questions were summed into a total knowledge score, and participants were divided into an adequate health knowledge group (score ≥ 6) or an inadequate health knowledge group (score < 5). Logistic regression was used determine the predictors of adequate CD health knowledge. The mean age of participants was 61.34 years (SD = 10 years). Out of a possible 12, the median score on the CD knowledge questionnaire was 3.0. About 25% of participants were classified as having adequate CD knowledge. Those who had a family history and/or long duration of CDs were more likely to have adequate health knowledge. Participants who received CD health information and self-care instructions from their physicians had 2.67 and 13.34 times greater odds of possessing adequate health knowledge than those who received no information, respectively. Adequate CD knowledge was strongly associated with regular check-ups, especially for those who attended township hospitals (OR = 40.17). Having regular check-ups at a fixed healthcare institution and receiving health information from physicians are important measures for increasing CD knowledge among rural adults with CDs. Township hospitals are the most effective settings for health education. It is important to develop an effective community-based prevention and control mechanism for CDs. This requires township hospitals to take a leading role in improving CD knowledge among chronically ill patients, and enhancing implementation of health education in rural China.


Steroidal contraception for men.
This article is a review of the development of male steroidal contraceptives during the past 25 years. Numerous studies have been conducted on male volunteers with oral and/or injectable preparations of single or combined steroids. Progestogen, androgen alone, or progestogen and androgen combinations have been used as weekly or monthly injectable formulations. Most of the studies involved small numbers of subjects. There was reversible suppression of spermatogenesis to oligospermia and/or azoospermia during the treatment period. Alteration of LH, FSH and testosterone levels in the blood was observed in most of these studies, depending on the steroid or combination of steroids used. There were reports about decreased or increased libido and weight gain during treatment with steroids. No other serious side-effects were found. Attention has recently focused on developing an androgen-only male contraceptive, because testosterone ester has shown promising results. The development of an effective and reliable steroidal contraceptive for men may be possible but this requires further research.


Regulation of telomerase activity by gamma-radiation in nasopharyngeal carcinoma cells.
Since telomerase has been suggested to play a role in radiation response, the effect of radiation on the regulation of telomerase activity was studied on two NPC cell lines, CNE1 and 915. In order to investigate the correlation between cell survival and telomerase activity, three low doses (0.25, 0.5 and 1 Gy) and three high doses (2, 4 and 8 Gy) of gamma-radiation were used. We found that low doses of radiation induced telomerase activity (up to 250% of untreated controls) followed by a cell cycle G1 arrest, suggesting that the up-regulation of telomerase may play a role in initiating radiation induced cell cycle arrest. High doses of radiation resulted in decreased telomerase activity (down to 30% of untreated controls) and this was followed by an increased cell death. Our results indicate that the up-regulation of telomerase activity is an important response to radiation induced sub-lethal DNA damage. Our results also suggest that inhibition of telomerase activity by high doses of radiation may play a role radiation-induced cell death.


Nature of serum ascorbate radical and its quantitative estimation.
A doublet signal was observed in human serum by the ESR technique at room temperature. This radical had a g value of 2.0054 and a hyperfine splitting constant of 1.84 gauss and was assigned to ascorbate radical. The ascorbate radical in serum was very stable. The intensity of ESR signal showed no differences between serum and plasma of the same individual. Photosensitivity of the ascorbate radical in serum and sodium ascorbate solution was examined and enhancement of ESR signal by irradiation was observed, although the responses in serum and in ascorbate solution were considerably different. The intensity of ESR signal was proportional to the concentration of ascorbate solution. The ESR intensity of ascorbate radical can be used as a reliable method for quantitative estimation of ascorbate radical.


Improving insomnia in primary care patients: A randomized controlled trial of nurse-led group treatment.
Insomnia is a common health problem, and most people who seek help for insomnia consult primary care. In primary care, insomnia treatment typically consists of hypnotic drugs, although cognitive behavioral therapy for insomnia is the recommended treatment. However, such treatment is currently available to few primary care patients. To evaluate the effects of a group treatment program for insomnia led by nurses in primary care. were the Insomnia Severity Index, a 2-week sleep diary, and a questionnaire on frequency of hypnotic drug use. A randomized controlled trial with pre- and post-treatment assessment and a 1-year post-treatment follow-up of the intervention group. Routine primary health care; 7 primary care centers in Stockholm, Sweden. Patients consulting primary care for insomnia were assessed for eligibility. To be included, patients had to have insomnia disorder and be 18 years or older. Patients were excluded if they if they worked night shifts or had severe untreated somatic and/or mental illness, bipolar disorder, or untreated sleep disorder other than insomnia. One-hundred and sixty-five patients 20 to 90 years were included. Most were women, and many had co-existing somatic and/or mental health problems. The post-treatment dropout rate was 20%. The intervention was a nurse-led group treatment for insomnia based on the techniques of cognitive behavioral therapy for insomnia. The nurses had 2days of training in how to deliver the program. Ninety patients were randomized to the intervention and 75 to the control group (treatment as usual). Data from 82 in the intervention and 71 in the control group were analyzed in accordance with intention-to-treat principles. Fifty-four of the 72 in the intervention group who participated in the group treatment program were followed up after 1year. Mean Insomnia Severity Index score decreased significantly from 18.4 to 10.7 after group treatment but remained unchanged after treatment as usual (17.0 to 16.6). The effect size was large (1.23). Group treatment also resulted in significant improvements in all sleep diary variables (sleep onset latency, total sleep time, time awake after sleep onset, number of awakenings, and sleep quality). It also reduced hypnotic drug use. Improvements were maintained 1-year post-treatment. Patients with insomnia can be treated successfully with a nurse-led group treatment program in primary health care. The results support implementation of the treatment program, particularly given the need for increased access to non-pharmacological insomnia treatments.


NOX4 Inhibition Potentiates Immunotherapy by Overcoming Cancer-Associated Fibroblast-Mediated CD8 T-cell Exclusion from Tumors.
Determining mechanisms of resistance to αPD-1/PD-L1 immune-checkpoint immunotherapy is key to developing new treatment strategies. Cancer-associated fibroblasts (CAF) have many tumor-promoting functions and promote immune evasion through multiple mechanisms, but as yet, no CAF-specific inhibitors are clinically available. Here we generated CAF-rich murine tumor models (TC1, MC38, and 4T1) to investigate how CAFs influence the immune microenvironment and affect response to different immunotherapy modalities [anticancer vaccination, TC1 (HPV E7 DNA vaccine), αPD-1, and MC38] and found that CAFs broadly suppressed response by specifically excluding CD8+ T cells from tumors (not CD4+ T cells or macrophages); CD8+ T-cell exclusion was similarly present in CAF-rich human tumors. RNA sequencing of CD8+ T cells from CAF-rich murine tumors and immunochemistry analysis of human tumors identified significant upregulation of CTLA-4 in the absence of other exhaustion markers; inhibiting CTLA-4 with a nondepleting antibody overcame the CD8+ T-cell exclusion effect without affecting Tregs. We then examined the potential for CAF targeting, focusing on the ROS-producing enzyme NOX4, which is upregulated by CAF in many human cancers, and compared this with TGFβ1 inhibition, a key regulator of the CAF phenotype. siRNA knockdown or pharmacologic inhibition [GKT137831 (Setanaxib)] of NOX4 "normalized" CAF to a quiescent phenotype and promoted intratumoral CD8+ T-cell infiltration, overcoming the exclusion effect; TGFβ1 inhibition could prevent, but not reverse, CAF differentiation. Finally, NOX4 inhibition restored immunotherapy response in CAF-rich tumors. These findings demonstrate that CAF-mediated immunotherapy resistance can be effectively overcome through NOX4 inhibition and could improve outcome in a broad range of cancers. SIGNIFICANCE: NOX4 is critical for maintaining the immune-suppressive CAF phenotype in tumors. Pharmacologic inhibition of NOX4 potentiates immunotherapy by overcoming CAF-mediated CD8+ T-cell exclusion. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/9/1846/F1.large.jpg.See related commentary by Hayward, p. 1799.


Native silica nanoparticles are powerful membrane disruptors.
Silica nanoparticles are under development for intracellular drug delivery applications but can also have cytotoxic effects including cell membrane damage. In this study, we investigated the interactions of silica nanospheres of different size, surface chemistry and biocoating with membranes of phosphatidylcholine lipids. In liposome leakage assays many, but not all, of these nanoparticles induced dose-dependent dye leakage, indicative of membrane perturbation. It was found that 200 and 500 nm native-silica, aminated and carboxylated nanospheres induce near-total dye release from zwitterionic phosphatidylcholine liposomes at a particle/liposome ratio of ∼1, regardless of their surface chemistry, which we interpret as particle-supported bilayer formation following a global rearrangement of the vesicular membrane. In contrast, 50 nm diameter native-silica nanospheres did not induce total dye leakage below a particle/liposome ratio of ∼8, whereas amination or carboxylation, respectively, strongly reduced or prevented dye release. We postulate that for the smaller nanospheres, strong silica-bilayer interactions are manifested as bilayer engulfment of membrane-adsorbed particles, with localized lipid depletion eventually leading to collapse of the vesicular membrane. Protein coating of the particles considerably reduced dye leakage and lipid bilayer coating prevented dye release all together, while the inclusion of 33% anionic lipids in the liposomes reduced dye leakage for both native-silica and aminated surfaces. These results, which are compared with the effect of polystyrene nanoparticles and other engineered nanomaterials on lipid bilayers, and which are discussed in relation to nanosilica-induced cell membrane damage and cytotoxicity, indicate that a native-silica nanoparticle surface chemistry is a particularly strong membrane interaction motif.


[The Dutch 'Wet op het Bevolkingsonderzoek' (Population Screening Act): adaptation preferable to repeal].
The Dutch 'Wet op het Bevolkingsonderzoek' (Population Screening Act) requires a license for some forms of screening, for example, screening for incurable diseases. In its recent report on commercial screening practices, the Dutch Health Inspectorate concludes that the Act is frequently not complied with. This is partly because in some respects the provisions of the Act are either not clear enough or difficult to apply. However, the more practical enforcement issues can and should be solved. Although some question whether the Act has a future, it should be remembered that the Act has demonstrated its value in the past. And in view of recent developments in the field of commercial screening and testing the need for legal instruments to control the increasing supply of screening services directly to the public is growing. A recent report by the Health Council of the Netherlands confirms this viewpoint.


Alcohol and marijuana use outcomes in the Healthy Choices motivational interviewing intervention for HIV-positive youth.
Healthy Choices is a motivational interviewing intervention targeting multiple risk behaviors among HIV-positive youth. This study investigated the effects of this intervention program specifically on alcohol and marijuana use. Youth living with HIV (n = 143, mean age = 20.7, 51.5% male) were recruited from four sites in the United States, and randomly assigned to intervention or control conditions. The four-session intervention focused on two of three possible problem behaviors based on entry screening; this study focused on 143 HIV-positive youth who received the intervention for substance use. At 15-month follow-up past-week alcohol use was significantly lower for intervention youth than control youth (39.7% versus 53.6%, χ2 = 2.81, 0.05 < p < 0.01); developmental trajectory analysis demonstrated significant reductions in alcohol use, but more importantly the intervention was effective over time in significantly reducing the adolescent's probability of being classified into the high-risk trajectory group. The intervention was less effective in reducing marijuana use.


Determinants of endocytic membrane geometry, stability, and scission.
During endocytic vesicle formation, distinct subdomains along the membrane invagination are specified by different proteins, which bend the membrane and drive scission. Bin-Amphiphysin-Rvs (BAR) and Fer-CIP4 homology-BAR (F-BAR) proteins can induce membrane curvature and have been suggested to facilitate membrane invagination and scission. Two F-BAR proteins, Syp1 and Bzz1, are found at budding yeast endocytic sites. Syp1 arrives early but departs from the endocytic site before formation of deep membrane invaginations and scission. Using genetic, spatiotemporal, and ultrastructural analyses, we demonstrate that Bzz1, the heterodimeric BAR domain protein Rvs161/167, actin polymerization, and the lipid phosphatase Sjl2 cooperate, each through a distinct mechanism, to induce membrane scission in yeast. Additionally, actin assembly and Rvs161/167 cooperate to drive formation of deep invaginations. Finally, we find that Bzz1, acting at the invagination base, stabilizes endocytic sites and functions with Rvs161/167, localized along the tubule, to achieve proper endocytic membrane geometry necessary for efficient scission. Together, our results reveal that dynamic interplay between a lipid phosphatase, actin assembly, and membrane-sculpting proteins leads to proper membrane shaping, tubule stabilization, and scission.


Races of Phytophthora sojae in Iran.
Phytophthora root and stem rot of soybean is a destructive disease of soybean in Iran. Races 1 and 3 of pathogen have already been reported from two major growing regions of the crop, Lorestan and Golestan provinces. In a survey during 2004-2005, 142 isolates of P. sojae were recovered from infected plants and naturally infested soil samples using selective media and soybean leaf baiting technique. The majority of tested isolates (110 isolates) belonged to race one of P. sojae and 32 isolates belonged to race 3. ITS region of 23 isolates were amplified with specific primers Ps1 and Ps2. Sequences of this regions were similar to other gene banks sequences except two isolates from China. This survey showed low diversity in Iranian population of P. sojae.


Cell quantification: evolution of compartmentalization and distribution of iron-oxide particles and labeled cells.
The purpose of the study was to show the feasibility of quantification in the case of cell death, cell migration and cell division by parametric MRI. We identify limitations for quantitative cell tracking owing to mixed parallel processes. Various intravoxel SPIO-labeled cell, super paramagnetic iron oxide particles (SPIO) and micron-sized paramagnetic iron oxide (MPIO) particle distributions were prepared by methods mimicking biologically relevant processes (compartmentalization, migration, division and cell death). R(2)* and R(2) relaxometry measurements were performed at 3.0 T; iron concentration was measured by optical emission spectrometry. The effects of spatial distribution and compartmentalization of paramagnetic iron-oxide particles on relaxivity were analyzed. Assessment of R(2)' (R(2)*-R(2)) allowed differentiation between intracellular and extracellular SPIO only if no high-iron-content extracellular particles were present. Relaxivity was sensitive to variations in cell labeling. Samples of the same cell types embedded in the same suspension media at the same cell density produced different relaxivity values, depending on the preparation of the labeled cells. In the case of cell division, a unique relationship between relaxation rate and iron concentration was found, where the relaxivity proved to be independent of initial cell labeling. In case of cell mixing, the cell density could be derived from relaxation values, even if iron concentration was undetermined. We demonstrated that relaxometry does not allow labeled cell quantification when multiple physiological processes such as cell division and cell migration coexist. The measured transversal relaxation rates were sensitive to the labeling technique. However, under special circumstances, despite the numerous limiting factors, quantification of the number of labeled cells by relaxometry was feasible.


A retrospective analysis of mortality risk and immunosuppressive therapy for Stevens-Johnson Syndrome and toxic epidermal necrolysis syndrome using the TriNetX research network.
Stevens-Johnson Syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) exist on a spectrum of autoimmune conditions which cause epidermal detachment and keratinocyte necrosis. Due to the rare incidence of these conditions, a dramatic heterogeneity in treatment algorithms exists. To better appreciate pharmacologic immunosuppressive therapies' impact on survival, the authors queried a multi-institutional data network. Data for this study was extracted from TriNetX Research Network, a platform that contains ICD-9/ICD-10 coding data from a consortium of international healthcare organizations. Seventy-one institutions were queried to identify adult patients diagnosed with SJS, TEN or SJS-TEN Overlap. Cohorts were created based on the therapy received: systemic steroids (SS), diphenhydramine (DH), cyclosporine (CS), intravenous immunoglobulin (IVIG), tumor necrosis factor alpha inhibitors (TNFαi), or a combination of treatments. Cohorts were then propensity matched with patients who received supportive care. Patients who only received one of the above treatments showed no significant reduction in 90-day mortality. Patients who received CS or IVIG as part of their multitherapy showed a significantly increased risk of death when compared to supportive care (CS: RR = 1.583, 95% CI [1.119, 2.240]; IVIG: RR = 2.132, 95% CI [1.485, 3.059]). Despite their frequent utilization, this study's analysis suggests that none of these therapies confer significant 90-day mortality survival over supportive care alone. These results highlight the heterogeneity of therapies and emphasize the need for critical prospective appraisal of their outcomes in SJS and TEN.


The effect of reducing dietary calcium in prestarter diets (0-4 D) on growth performance of broiler chickens, tibia characteristics, and calcium and phosphorus concentration in blood.
During the incubation period, the Ca-to-P weight (mg/mg) ratio in the yolk increases from 0.26 on day 0 to 0.92 on day 17.5 and to 2.9 at hatch. Moreover, the absolute Ca content in the yolk increases by 41%, whereas P content decreases by 87%, from day 0 to the day of hatching. Thus, at hatch and during the first days after hatching, there are high reserves of Ca relative to P in the residual egg yolk, risking hypercalcemia and hypophosphatemia. A growth performance study was conducted to explore the effects of reducing dietary Ca content in the prestarter phase (0-4 D) on BW and bone mineral deposition during the first days after hatch and at market weight (day 37). Four prestarter (0-4 D) diets were formulated to have 0.4, 0.6, 0.8, and 1.0% Ca content. After the prestarter phase, all birds were fed with the same commercial diets based on a 3-phase feeding program (starter, grower, and finisher). Growth performance (BW, ADG, ADFI, and feed conversion ratio [FCR]) was monitored throughout the study, and blood and tibia bone samples were collected on specific days. On day 4, BW and ADG decreased with dietary Ca contents higher than 0.6% (P < 0.05), but there were no differences in BW on day 14 onward (P > 0.10). For the overall study (0-37 D), there were no differences in ADG and ADFI, but the FCR decreased with lower Ca contents (P < 0.05). On day 4, there were no differences in blood plasma Ca concentration, but P concentration increased in the group treated with diet containing 0.4% Ca compared with the groups treated with diets containing 0.6 and 0.8% Ca (P < 0.05). Tibia ash content decreased in the group treated with diet containing 0.4% Ca (P < 0.05) compared with all other treatments at the end of the prestarter phase. Tibia ash content and tibia breaking strength, on day 37, were not different among the treatments (P > 0.10). In conclusion, during the prestarter phase, BW increased with dietary Ca contents lower than 0.6%, most likely improving Ca-P balance; bone mineral deposition was reduced in this period. On feeding with a diet containing higher Ca content, bone mineral content was rapidly recovered.


One-bead one-compound combinatorial library derived targeting ligands for detection and treatment of oral squamous cancer.
Oral squamous cancers (OSC) are hallmarked by poor prognosis, delayed clinical detection, and a lack of defined, characteristic biomarkers. By screening combinatorial one-bead one-compound (OBOC) peptide libraries against oral squamous cancer cell lines, two cyclic peptide ligands, LLY12 and LLY13 were previously identified. These ligands are capable of specific binding to the oral cancer cell lines (MOK-101, HSC-3, SCC-4 and SCC-10a) but not non-cancerous keratinocytes, leukocytes, fibroblast, and endothelial cells. These two peptides were synthesized and evaluated for their binding property, cytotoxicity and cell permeability. In vitro studies indicate that both LLY12 and LLY13 were able to bind to oral cancer cells with high specificity but did not show any cytotoxicity against human keratinocytes. Biotinylated LLY13, in complex with streptavidin-alexa488 was taken up by live oral cancer cells, thus rendering it as an excellent candidate vehicle for efficient delivery of drug loaded-nanoparticles. In vivo and ex vivo near infra-red fluorescence imaging studies confirmed the in vivo targeting efficiency and specificity of LLY13 in oral cancer orthotopic murine xenograft model. In vivo studies also showed that LLY13 was able to accumulate in the OSC tumors and demarcate the tumor margins in orthotopic xenograft model. Together, our data supports LLY13 as a promising theranostic agent against OSC.


Heart transplantation--an autopsy review.
Cardiac transplantation is now a well recognised form of treatment for several forms of end stage cardiac failure, particularly ischaemic heart disease and cardiomyopathies. By the end of 1988, seven patients from Northern Ireland had received cardiac transplants in Papworth and Harefield Hospitals. Of these, four have died, three as a result of rejection and the fourth from widespread cytomegalovirus infection. The autopsy findings in these four cases are presented in this paper.


Inbreeding depression in Solanum carolinense (Solanaceae) under field conditions and implications for mating system evolution.
The clonal weed Solanum carolinense exhibits plasticity in the strength of its self-incompatibility (SI) system and suffers low levels of inbreeding depression (δ) in the greenhouse. We planted one inbred and one outbred plant from each of eight maternal plants in a ring (replicated twice) and monitored clonal growth, herbivory, and reproduction over two years. Per ramet δ was estimated to be 0.63 in year one and 0.79 in year two, and outbred plants produced 2.5 times more ramets than inbred plants in the spring of year two. Inbred plants also suffered more herbivore damage than outbred plants in both fields, suggesting that inbreeding compromises herbivore resistance. Total per genet δ was 0.85 over the two years, indicating that S. carolinense is unlikely to become completely self-compatible, and suggesting that plasticity in the SI system is part of a stable mixed-mating system permitting self-fertilization when cross pollen limits seed production.


First-Principles Study of Chemical Mixtures of CaCl2 and MgCl2 Hydrates for Optimized Seasonal Heat Storage.
Chloride-based salt hydrates form a promising class of thermochemical materials (TCMs), having high storage capacity and fast kinetics. In the charging cycles of these hydrates however hydrolysis might appear along with dehydration. The HCl produced during the hydrolysis degrades and corrodes the storage system. Our GGA-DFT results show that the enthalpy charge during proton formation (an important step in hydrolysis) is much higher for CaCl2·2H2O (33.75 kcal/mol) than for MgCl2·2H2O (19.55 kcal/mol). This is a strong indicator that hydrolysis can be minimized by appropriate chemical mixing of CaCl2 and Mg Cl2 hydrates, which is also confirmed by recent experimental studies. GGA-DFT calculations were performed to obtain and analyze the optimized structures, charge distributions, bonding indicators and harmonic frequencies of various chemical mixtures hydrates and compared them to their elementary salts hydrates. We have further assessed the equilibrium products concentration of dehydration/hydrolysis of the chemical mixtures under a wide range of operating conditions. We observed that chemical mixing leads to an increase of the onset hydrolysis temperature with a maximum value of 79 K, thus increasing the resistance against hydrolysis with respect to the elementary salt hydrates. We also found that the chemical mixing of CaCl2 and MgCl2 hydrates widens the operating dehydration temperature range by a maximum value of 182 K (CaMg2Cl6·2H2O) and lowers the binding enthalpy with respect to the physical mixture by ≈65 kcal/mol for TCM based heat storage systems.


Adherence to the MIND diet and the risk of cardiovascular disease in adults: a cohort study.
The Mediterranean-DASH intervention for neurodegenerative delay (MIND) and cardiovascular disease (CVD) has yet to be studied. As a result, the objective of this study was to investigate if there was a link between MIND diet adherence and the risk of CVD in adults over a 10.6-year period of time. There were 2863 participants in this research cohort who were free of CVD and were followed-up for 10.6 years. Using a validated food frequency questionnaire (FFQ), the MIND diet score was calculated. Reliable medical data was used to evaluate cardiovascular events, including coronary heart disease (CHD), stroke, and CVD-related mortality. A total of 200 CVD occurrences (159 CHD, 21 stroke, and 20 CVD fatalities) were documented throughout the 10.6-year follow-up period. Our results indicated each increase in MIND diet score reduced the incidence of CVD by 16% (HR: 0.84; 95% CI: 0.74 to 0.96). We also detected an inverse association relationship between the MIND diet and the incidence of CVD after classifying the MIND diet score into tertiles as well (HR: 0.68; 95% CI: 0.47 to 0.97). Among the components of the MIND diet, each increase in whole grains, green leafy vegetables, and beans reduced the risk of CVD by 60%, 45%, and 65%, respectively. Finally, our findings revealed that the higher adherence to the MIND was associated with a lower risk of cardiovascular events. Besides, we found an inverse association between consumption of whole grains, green leafy vegetables, and beans and the number of CVD incidents. However, more well-designed cohort studies are needed before conclusive conclusions can be formed.


The radiating part of circular sources.
An analysis is developed linking the form of the sound field from a circular source to the radial structure of the source, without recourse to far-field or other approximations. It is found that the information radiated into the field is limited, with the limit fixed by the wavenumber of the source multiplied by the source radius (Helmholtz number). The acoustic field is found in terms of the elementary fields generated by a set of line sources whose form is given by Chebyshev polynomials of the second kind and whose amplitude is found to be given by weighted integrals of the radial source term. The analysis is developed for tonal sources, such as rotors, and for Helmholtz number less than two, for random disk sources. In this case, the analysis yields the cross-spectrum between two points in the acoustic field. The analysis is applied to the problems of tonal radiation, random source radiation as a model problem for jet noise, and to noise cancellation, as in active control of noise from rotors. It is found that the approach gives an accurate model for the radiation problem and explicitly identifies those parts of a source which radiate.


Physical Chemistry of Chloroquine Permeation through the Cell Membrane with Atomistic Detail.
We provide a molecular-level description of the thermodynamics and mechanistic aspects of drug permeation through the cell membrane. As a case study, we considered the antimalaria FDA approved drug chloroquine. Molecular dynamics simulations of the molecule (in its neutral and protonated form) were performed in the presence of different lipid bilayers, with the aim of uncovering key aspects of the permeation process, a fundamental step for the drug's action. Free energy values obtained by well-tempered metadynamics simulations suggest that the neutral form is the only permeating protomer, consistent with experimental data. H-bond interactions of the drug with water molecules and membrane headgroups play a crucial role for permeation. The presence of the transmembrane potential, investigated here for the first time in a drug permeation study, does not qualitatively affect these conclusions.


Na(+)/Ca(2+) exchanger in GABAergic presynaptic boutons of rat central neurons.
Rat Meynert neurons were acutely isolated using a dissociation technique that maintains functional GABAergic presynaptic boutons. Miniature inhibitory postsynaptic currents (mIPSCs) were recorded under voltage-clamp conditions using whole cell patch-clamp recordings. Using the frequency of mIPSCs as a measure of presynaptic terminal excitability, the existence of a Na(+)/Ca(2+) exchanger (NCX) in these GABAergic nerve terminals was clearly demonstrated. Both the frequency and the amplitude of mIPSCs were unaffected by replacement of extracellular Na(2+). However, in this Na(+)-free external solution, ouabain could now induce a transient increase of mIPSCs frequency, which was not inhibited by adding Cd(2+) or cyclopiazonic acid but was inhibited by removing external Ca(2+). This indicates that this transient potentiation was dependent on external Ca(2+), but that this Ca(2+) influx was not via voltage-dependent Ca(2+) channels. KB-R7943, an inhibitor of NCX, at a concentration of 3 x 10(-6) M, reduced this transient increase of mIPSCs frequency without affecting mIPSCs amplitude and the response to exogenous GABA. These results demonstrate the existence of NCX in these GABAergic nerve terminals. In zero external Na(+), ouabain causes an accumulation of intraterminal Na(+) and a resultant influx of Ca(2+) through the reversed mode operation of NCX. However, under more physiological conditions, NCX may also operate in a forward mode and serve to maintain low intracellular [Ca(2+)] in nerve terminals.


Combined evaluation of urokinase-type plasminogen activator and plasminogen activator inhibitor-2 expression in gastric carcinoma.
Urokinase-type plasminogen activator (UPA) is a serine protease implicated in cancer invasion and metastasis. There are at least two kinds of specific inhibitor which act on UPA: plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2). It has been reported that both UPA and PAI-1 antigen levels are associated with short survival in patients with breast cancer. In this study, we investigated the correlation between the expression of UPA, PAI-2 and prognosis in gastric carcinoma. One hundred and twenty specimens resected from patients with gastric carcinoma were investigated by staining with monoclonal antibodies against UPA and PAI-2. UPA positive rate was significantly higher in patients with liver metastases than in those without such metastases, There was no significant association between PAI-2 expression and clinicopathologic factors. However, patients with UPA-positive and PAI-2-negative tumors had more advanced cancer than other patients. According to the prognosis, the patients with UPA-positive and PAI-2-negative tumors had a significantly poorer prognosis than other patients. In conclusion, the combined evaluation of UPA and PAI-2 expression is associated with tumor progression in gastric cancer.


Exploring the effectiveness of dextroamphetamine for the treatment of stimulant use disorder: a qualitative study with patients receiving injectable opioid agonist treatment.
A high proportion of people receiving both oral and injectable opioid agonist treatment report concurrent use of stimulants (i.e. cocaine and or amphetamines), which has been associated with higher rates of continued illicit opioid use and treatment dropout. A recent randomized controlled trial demonstrated the effectiveness of dextroamphetamine (a prescribed stimulant) at reducing craving for and use of cocaine among patients receiving injectable opioid agonist treatment. Following this evidence, dextroamphetamine has been prescribed to patients with stimulant use disorder at a clinic in Vancouver. This study investigates perceptions of the effectiveness of dextroamphetamine from the perspective of these patients. Data were collected using small focus groups and one-on-one interviews with patients who were currently or formerly receiving dextroamphetamine (n = 20). Thematic analysis was conducted using an iterative approach, moving between data collection and analysis to search for patterns in the data across transcripts. This process led to the defining and naming of three central themes responding to the research question. Participants reported a range of stimulant use types, including cocaine (n = 8), methamphetamine (n = 8), or both (n = 4). Three central themes were identified as relating to participants' perceptions of the effectiveness of the medication: 1) achieving a substitution effect (i.e. extent to which dextroamphetamine provided a substitution for the effect they received from use of illicit stimulants); 2) Reaching a preferred dose (i.e. speed of titration and effect of the dose received); and 3) Ease of medication access (i.e. preference for take home doses (i.e. carries) vs. medication integrated into care at the clinic). In the context of continued investigation of pharmacological treatments for stimulant use disorder, the present study has highlighted how the study of clinical outcomes could be extended to account for factors that contribute to perceptions of effectiveness from the perspective of patients. In practice, elements of treatment delivery (e.g. dosing and dispensation protocols) can be adjusted to allow for various scenarios (e.g. on site vs. take home dosing) by which dextroamphetamine and other pharmacological stimulants could be implemented to provide "effective" treatment for people with a wide range of treatment goals and needs.


Ultrasound may decrease the emergency surgery rate of incarcerated inguinal hernia.
Manual reduction is the standard procedure for incarcerated inguinal hernia reduction. The role of ultrasound in incarcerated inguinal hernia reduction has not been defined. The aim of this study was to determine whether ultrasound can improve the ability to reduce incarcerated inguinal hernia safely when manual reduction fails and thereby decrease the emergency surgery rate. Between January 1994 and December 2003, 112 adult patients with incarcerated inguinal hernias were admitted to a university medical center and classified into two groups. Group I consisted of 61 patients who received emergency surgical reduction after the failure of two attempts at manual reduction. In Group II, comprising 51 patients, ultrasound-guided reduction was performed when two attempts at manual reduction failed. Emergency surgical reduction was scheduled if both ultrasound-guided reduction and two attempts at manual reduction failed. The difference in emergency surgery rates between groups I and II was compared. In group I, manual reduction was successful in 55 cases (90.2%) and 6 patients underwent emergency surgery. In group II, manual reduction was successful in 45 cases, ultrasound-guided reduction in 4 cases, 1 reduction was avoided after ultrasonic examination, and 1 patient underwent emergency surgery. The emergency surgery rates in groups I and II were 9.8% and 2.0%, respectively. Ultrasound can improve the ability to reduce incarcerated inguinal hernia safely when manual reduction fails and may decrease the rate of emergency surgery.


Improved algorithms for automatic bone histomorphometry on a numerized image analysis system.
Quantitative histological methods have proved to be the most effective methods in bone disease research. Faster and more accurate techniques are currently needed. We have developed a simple digitized image analysis system which allows accurate measurements of trabecular bone mass. The algorithm is based on the 'four-connected sets' mathematical theory. Given a numerized image displayed by a CCD camera, the algorithm recognizes all possible four-connected sets and provides area measurements. The first procedure automatically eliminates small, irrelevant profiles (wrinkles, cell nuclei, etc.) while larger profiles are erased interactively. The second procedure similarly erases the artefactual defects within the trabeculae (artefactual cracks or empty osteocytic lacunae). The method was shown to be very accurate and time-saving.


Agroinfiltration reduces ABA levels and suppresses Pseudomonas syringae-elicited salicylic acid production in Nicotiana tabacum.
Agrobacterium tumefaciens strain GV3101 (pMP90) is widely used in transient gene expression assays, including assays to study pathogen effectors and plant disease resistance mechanisms. However, inoculation of A. tumefaciens GV3101 into Nicotiana tabacum (tobacco) leaves prior to infiltration with pathogenic and non-host strains of Pseudomonas syringae results in suppression of macroscopic symptoms when compared with leaves pre-treated with a buffer control. To gain further insight into the mechanistic basis of symptom suppression by A. tumefaciens we examined the effect of pre-treatment with A. tumefaciens on the growth of P. syringae, the production of the plant signalling molecules salicylic acid (SA) and abscisic acid (ABA), and the presence of callose deposits. Pre-treatment with A. tumefaciens reduced ABA levels, P. syringae multiplication and P. syringae-elicited SA and ABA production, but promoted increased callose deposition. However, pre-treatment with A. tumefaciens did not suppress necrosis or SA production in leaves inoculated with the elicitor HrpZ. Collectively, these results show that inoculation of N. tabacum leaves with A. tumefaciens alters plant hormone levels and plant defence responses to P. syringae, and demonstrate that researchers should consider the impact of A. tumefaciens on plant signal transduction when using A. tumefaciens-mediated transient expression assays to investigate ABA-regulated processes or pathogenicity and plant defence mechanisms.


Spine Oncology - Primary Spine Tumors.
Primary tumors originating from the spine are very complex and challenging entities to treat. Due to their rarity, a multicenter collaborative network is essential to shepherd the best research and contribute to the dissemination of the best evidence possible. Over the last few years, several advances have occurred in many different fields. Surgery is still the cornerstone of treatment in most cases. The occasional suboptimal outcomes and high morbidity of surgical treatment have however encouraged professionals caring for these patients to explore safer treatment options and alternatives or adjuncts to surgical treatment. A number of novel treatment strategies have emerged from the medical, interventional radiology, radiation oncology, and molecular worlds. This has truly positioned primary spine tumors at the forefront of multidisciplinary care. This article discusses these recent advances in detail to equip the oncologic spine surgeon and their team to better counsel and treat these patients. Most of these advances allow for a more tailored, efficient, and, most importantly, less morbid management of primary spine tumors. Some of these advances are still under investigation, however, and evidence-based oncological principles should still be strongly encouraged.


Microwave detection of metastasized breast cancer cells in the lymph node; potential application for sentinel lymphadenectomy.
Metastasis is the leading cause of death in breast cancer patients and an appropriate detection of metastasis can provide better prognosis and quality treatments. Microwaves can reveal the unique electromagnetic properties of materials, and this study aims to unleash the electromagnetic properties of breast cancer cells, especially, metastasized cancer cells in the lymph nodes, using broad-band microwaves in attempts to detect metastases. To distinguish the cancer-specific patterns of cancer tissues, three primary microwave parameters were assessed, i.e., permittivity in mid-band frequency (3-5 GHz), conductivity in high-band frequencies (25-30 GHz) and slope changes of permittivity at high-band frequencies (15-30 GHz). An additional parameter, Cancer Metastasis Index (CMI), was developed to effectively represent all parameters. Broadband microwave scanning can reveal cancer specific electromagnetic behaviors in all three parameters, and these were reliably reflected by CMI. CMI effectively magnified the difference of the electromagnetic properties between normal nodal tissues and cancer tissues. immunohistochemistries were performed to verify the origin of electromagnetic changes represented by CMI values.


In vitro dissolution models for the prediction of in vivo performance of an oral mesoporous silica formulation.
Drug release from mesoporous silica systems has been widely investigated in vitro using USP Type II (paddle) dissolution apparatus. However, it is not clear if the observed enhanced in vitro dissolution can forecast drug bioavailability in vivo. In this study, the ability of different in vitro dissolution models to predict in vivo oral bioavailability in a pig model was examined. The fenofibrate-loaded mesoporous silica formulation was compared directly to a commercial reference product, Lipantil Supra®. Three in vitro dissolution methods were considered; USP Type II (paddle) apparatus, USP Type IV (flow-through cell) apparatus and a USP IV Transfer model (incorporating a SGF to FaSSIF-V2 media transfer). In silico modelling, using a physiologically based pharmacokinetic modelling and simulation software package (Gastroplus™), to generate in vitro/in vivo relationships, was also investigated. The study demonstrates that the in vitro dissolution performance of a mesoporous silica formulation varies depending on the dissolution apparatus utilised and experimental design. The findings show that the USP IV transfer model was the best predictor of in vivo bioavailability. The USP Type II (paddle) apparatus was not effective at forecasting in vivo behaviour. This observation is likely due to hydrodynamic differences between the two apparatus and the ability of the transfer model to better simulate gastrointestinal transit. The transfer model is advantageous in forecasting in vivo behaviour for formulations which promote drug supersaturation and as a result are prone to precipitation to a more energetically favourable, less soluble form. The USP IV transfer model could prove useful in future mesoporous silica formulation development. In silico modelling has the potential to assist in this process. However, further investigation is required to overcome the limitations of the model for solubility enhancing formulations.


[Immunochemical studies of salivary lactoferrin].
Immunodiffusion assays were used to examine lactoferrin (LF) in the mixed saliva of 1026 patients and 553 apparently healthy individuals. Elevated salivary LF levels were found in respiratory and digestive diseases and diabetes. It is concluded that this non-invasive immunochemical assay is promising in the screening and monitoring of persons during their health examination.


[Surgical site infection and its risk factors in colon surgeries].
To identify the occurrence of surgical site infection (SSI) and its risk factors in patients undergoing colon surgery in a tertiary hospital located in the countryside of the state of São Paulo. Retrospective cohort study, with collection of information contained in the medical records of patients undergoing colon surgery in the period between January 2010 and December 2013. The studied variables were the possible risk factors related to the patient, to demographic characteristics and the surgical procedure. In total, were evaluated 155 patients with an overall SSI incidence of 16.7%. A statistically significant association was found both in the univariate as in the multivariate analysis between the SSI and the following variables: male gender, Charlson index and mechanical bowel preparation. The understanding of health professionals about the factors that influence the incidence of SSI in colon surgery may contribute to the quality of care provided to surgical patients, from effective actions to minimize the risk of infections.


Dose-Dependent Effects of the Cimicifuga racemosa Extract Ze 450 in the Treatment of Climacteric Complaints: A Randomized, Placebo-Controlled Study.
Extracts from Cimicifuga racemosa (CR, synonym Actaea racemosa) have shown efficacy in trials in women with menopausal symptoms. Yet, dose dependency remains unclear. Therefore, 180 female outpatients with climacteric complaints were treated for 12 weeks in a randomized, double-blind, placebo-controlled, 3-armed trial (CR extract Ze 450 in 6.5 mg or 13.0 mg, or placebo). Primary outcome was the difference in menopausal symptoms (vasomotor, psychological, and somatic), assessed by the Kupperman Menopausal Index between baseline and week 12. Secondary efficacy variables were patients' self-assessments of general quality of life (QoL), responder rates, and safety. Compared to placebo, patients receiving Ze 450 showed a significant reduction in the severity of menopausal symptoms in a dose-dependent manner from baseline to endpoint (mean absolute differences 17.0 (95% CI 14.65-19.35) score points, P < 0.0001 for 13.0 mg; mean absolute differences 8.47 (95% CI 5.55-11.39) score points, P = 0.0003 for 6.5 mg). QoL and responder rates corresponded with the main endpoint. Changes in menopausal symptoms and QoL were inversely correlated. Reported adverse events and clinical laboratory testing did not raise safety concerns. The CR extract Ze 450 is an effective and well-tolerated nonhormonal alternative to hormone treatment for symptom relief in menopausal women.


Kinetic stability of the peroxidase activity of unfolded cytochrome c: heme degradation and catalyst inactivation by hydrogen peroxide.
Unfolding converts Paracoccus versutus cytochrome c-550 into a potent peroxidase (Diederix, R. E. M.; Ubbink, M.; Canters, G. W. ChemBioChem 2002, 3, 110-112). The catalytic activity is accompanied by peroxide-driven inactivation that is prevented, in part, by reducing substrate. Here, the kinetics of inactivation are described, and evidence is presented for the occurrence of a labile intermediate on the catalytic peroxidase pathway of unfolded cytochrome c-550. This intermediate represents a branching point, whereby the protein proceeds along either the productive pathway or self-inactivates. Reducing substrate suppresses inactivation by decreasing the steady-state concentration of the labile intermediate. Inactivation is accompanied by heme degradation. Its chemical reactivity, UV-vis, and EPR properties identify the first intermediate as hydroxyheme-cytochrome c-550, i.e. with heme hydroxylated at one of the heme meso positions. The occurrence of this species argues for the peroxo-iron species in the peroxidase mechanism as the labile intermediate leading to inactivated cytochrome c-550.


[Longitudinality in nurses' work: a report of professional experiences].
The present study aimed at describing nurses' perceptions regarding care under the perspective of longitudinality. This descriptive-exploratory qualitative study was performed in April 2010 with twenty family health nurses from cities of the 10th Regional Health District of Paraná, using semi-structured interviews. The data were submitted to content analysis. Results pointed at the following category: benefits and implications of interventions from a perspective of longitudinality, which found that the referenced care occurs with children, clients with chronic disease, families, and through groups. Teamwork, accessibility, client's co-participation, home visits and health prevention actions make longitudinality feasible and have a positive impact on peoples' health. In conclusion, longitudinality improves the quality of life of the population and promotes resolvability in primary healthcare.


Early drinking onset: a study of prevalence and determinants among 13-year-old adolescents in Norway.
Early drinking onset is associated with different psychosocial adjustment problems among adolescents. The aim of this study was to assess determinants associated with early drinking and to identify factors predicting early drinking onset among adolescents. The study included 1,550 eighth-graders with a mean age of 13.5 years from 41 schools. A total of 24% (boys 29%, girls 19%) had ever drunk alcohol, while 14% had drunk some alcohol in the last 30 days. Further, early drinking was associated with gender, religion, school performance, smoking and bullying in the bivariate tests. Predictors of early drinking onset were identified by generalized linear mixed models with two multivariable models created. The first model included social and environmental variables. Entering intentions, expectancies, attitudes and norms into the multivariable analysis resulted in a significant improvement of the model fit constituting 86% in the second model. The percentage correctly classified those (56%) who had been drinking in the second model which was two times higher compared to the first model. Gender, religion and smoking emerged as significant predictors of drinking in both models.


Differences in expression of transcription factor AP-1 in human promyelocytic HL-60 cells during differentiation towards macrophages versus granulocytes.
Commitment of HL-60 cells to macrophage or granulocytic differentiation was achieved by incubation with 4 beta-phorbol 12-myristate 13-acetate (PMA) for 30-60 min or with dimethyl sulphoxide (DMSO) for 24 h respectively. The commitment stage towards PMA-induced macrophage differentiation was associated with increases in jun B and c-fos mRNA levels, as well as with an increase in the binding activity of transcription factor AP-1. Nevertheless, gel retardation analysis indicated that the AP-1 activity detected in untreated cells was drastically reduced during the commitment stage of DMSO-induced HL-60 differentiation towards granulocytes. When HL-60 cells were treated with sodium butyrate, which induced monocytic differentiation, a remarkable increase in AP-1 binding activity was detected. Treatment of HL-60 cells with 1 alpha,25-dihydroxyvitamin D3, another monocytic differentiation agent, induced a weak, but appreciable, increase in AP-1 activity. Furthermore, addition of sodium butyrate or 1 alpha,25-dihydroxyvitamin D3 to HL-60 cells induced the expression of c-fos, c-jun, jun B and jun D proto-oncogenes. In contrast, when HL-60 cells were treated with retinoic acid, a granulocytic differentiation inducer, no enhanced AP-1 binding activity was observed, and only a weak increase in jun D mRNA level was detected. These data indicate that formation of AP-1 is not required for the induction of HL-60 differentiation towards granulocytes, whereas induction of monocytic differentiation is correlated with an increase in AP-1 activity. The differential expression of AP-1 activity may be critical in the differentiation of HL-60 cells towards monocytic or granulocytic lineages.


Identification of children at risk of Attention Deficit/Hyperactivity Disorder: a school-based intervention.
In many countries, the majority of children who have Attention Deficit/Hyperactivity Disorder (ADHD) are undiagnosed and there is limited recognition of child mental health problems in primary care. Teachers may be well placed to identify unrecognised children and to facilitate their referral to specialist services. Despite this, there has been limited intervention research addressing teacher identification of ADHD. This study aims to examine whether an educational intervention about ADHD for teachers improves their recognition of children at risk of ADHD. A before and after investigation of an educational intervention about ADHD for teachers took place in 6 primary schools (involving 96 class teachers and 2672 pupils). Teacher recognition was compared against a diagnostic algorithm for ADHD caseness that utilised both parent and teacher ratings. Changes in teacher recognition of children with probable ADHD, as well as predictors of recognition, were examined. Following the intervention, there was an increase in the proportion of children regarded by teachers as having probable ADHD. There was also improved agreement between teacher recognition and the diagnostic algorithm. Teacher views that a child had probable ADHD were based on both the severity of symptoms and the impact of these problems on the teacher and the class. It is feasible to deliver an educational intervention addressing teacher identification of ADHD in routine practice. This was associated with an improvement in the ability of teachers to more accurately identify children at risk of ADHD. The provision of a brief educational intervention for teachers could assist in improving the identification of undiagnosed children with ADHD in the community.


Detection of human cytomegalovirus pp67 late gene transcripts in cerebrospinal fluid of human immunodeficiency virus type 1-infected patients by nucleic acid sequence-based amplification.
This study examined the clinical correlation between the presence of human cytomegalovirus (HCMV) pp67 mRNA in cerebrospinal fluid (CSF) and active HCMV central nervous system (CNS) disease in patients with human immunodeficiency virus type 1 (HIV-1). In total, 76 CSF specimens collected from 65 HIV-1-positive patients diagnosed with HCMV CNS disease, other non-HCMV-related CNS diseases, or no CNS disease were tested for the presence of HCMV pp67 mRNA using the NucliSens cytomegalovirus (CMV) pp67 assay (Organon Teknika, Durham, N.C.). The results were compared to those of a nested PCR for the detection of HCMV glycoprotein B DNA and to those obtained by viral culture (54 samples). CSF specimens collected from patients without HCMV CNS disease yielded the following results: pp67 assay negative, 62 of 62 specimens; culture negative, 41 of 41 specimens; and PCR negative, 56 of 62 specimens (6 specimens were positive). CSF specimens collected from patients with HCMV CNS disease yielded the following results: pp67 assay positive, 9 of 13 specimens; PCR positive, 13 of 13 specimens; and culture positive, 2 of 13 specimens. After resolution of the discordant results, the following positive and negative predictive values (PPV and NPV, respectively) for the diagnosis of HCMV CNS disease were determined. The PPV for PCR, pp67 assay, and culture were 68.4, 100, and 100%, respectively, and the NPV for PCR, pp67 assay, and culture were 100, 97.0, and 82. 7%, respectively. The sensitivities for DNA PCR, pp67 assay, and culture for the detection of HCMV were 100, 84.6, and 18%, respectively, and the clinical specificities were 90.5, 100, and 100%, respectively. This study indicates that the detection of HCMV pp67 mRNA in CSF has good correlation with active HCMV CNS disease, whereas CSF culture is insensitive and qualitative DNA PCR may detect latent nonreplicating virus in CSF from patients without HCMV CNS disease.