Promoting Healthy Ageing in South Africa Through Vaccination of the Elderly.
The World Health Organization estimates that globally, the proportion of people aged ≥60 years will more than double by the year 2050, with the majority of elderly people living in low- and middle-income countries such as South Africa. Population ageing is an impending public health concern, potentially negatively impacting on South Africa's economy and health system if the government does not adequately prepare for this change. Globally, many potential solutions to ensure healthy ageing are being discussed and implemented, including adopting a "life-course" approach to vaccination which includes vaccination of the elderly, since they are at considerable risk of severe morbidity and mortality from vaccine-preventable diseases. While vaccines are considered as one of the greatest tools for preventing childhood infectious disease morbidity and mortality, they are under-utilised in strategies for promoting healthy ageing in South Africa, where only influenza vaccination is available free of charge to the elderly accessing public sector healthcare. Population ageing coupled with the high incidence of vaccine-preventable diseases amongst elderly South Africans, necessitates establishing a comprehensive national policy and guidelines for vaccination of the elderly.


Resourse Use and Disease Couse in dementia - Nursing Home (REDIC-NH), a longitudinal cohort study; design and patient characteristics at admission to Norwegian nursing homes.
Earlier studies of nursing home patients show a high prevalence of dementia, neuropsychiatric symptoms (NPS), pain, and dependency in activities of daily living. The REDIC-NH cohort was set up to study the disease course and the resources used in patients with dementia in Norway. The aim of this paper was to describe the methods and the data collection, and to present selected data about patients at admission to a nursing home. We included 696 patients at admission to a nursing home and followed them with biannual assessments until death. Baseline data were collected between March 2012 and November 2014. In October 2016, patients had either completed an 18-month follow-up (n = 349), passed 18 months without assessments (n = 22), or left the study (n = 324). Data on demographics, cognition, NPS, activities of daily living (ADL) functioning, physical health, medication, Quality of Life (QoL), resource use, and caregiver burden, in addition to DNA samples were collected. Mean age of the participants at inclusion was 84.5 years (SD 7.5, range 50 - 105), 63.9% were women. According to data collected in the study, 83.8% had dementia, but only 55.9% of them had a diagnosis of dementia registered in their records. The most frequent dementia diagnosis was Alzheimer's disease, which was present in 71% of those with dementia. Patients with dementia more often experienced delusions, hallucinations, agitation, anxiety, disinhibition, irritability, and aberrant motor behaviour compared to patients without dementia. Depression and anxiety were the most common NPS symptoms. Dementia and NPS were highly prevalent among persons admitted to nursing homes. Only 55.9% of the patients with dementia had a diagnosis of dementia registered in their records.


Anatomic correction of simple transposition of the great arteries in 50 neonates.
From April 1984 to January 1986, anatomic surgical correction was performed in 50 newborn (2- to 23-day-old, mean 8 +/- 5 [SD]) infants with simple transposition of the great arteries. Before surgery, balloon atrial septostomy was performed in all patients, prostaglandin E1 was infused in 42, and left ventricular shape on a two-dimensional echocardiogram was considered "satisfactory" in 48. Surgery was performed in patients on cardiopulmonary bypass without cardiac arrest; the pulmonary artery was reconstructed by end-to-end anastomosis according to Lecompte's maneuver with a pericardial patch. In all but one patient coronary artery transfer was possible regardless of the distribution of these vessels. There were eight early deaths (16%), but only four (10%) of the last 41 patients treated died. There was one late death (2%) due to a secondary myocardial infarction caused by compression of the left coronary artery. Reoperation was successfully performed in two patients for supravalvar pulmonary artery stenosis. The only late medical complication was a transient episode of myocardial ischemia 6 months after surgery. The 41 late survivors were in excellent condition, were in sinus rhythm, and had a normal left ventricle 1 to 22 months after surgery (mean 7.2 +/- 5.4 [SD]). Aortic growth was normal; pulmonary artery supravalvar stenosis occurred in six patients (mild in four). We conclude that anatomic correction can be applied successfully in the first few days of life in newborns with simple transposition of the great arteries, regardless of coronary distribution.


PAMPs and DAMPs in allergy exacerbation models.
Sensitization of mice to real-life allergens or harmless antigen with the use of adjuvants will lead to the induction of DAMPs in the immune system. We have shown that the Th2-inducing adjuvant aluminum hydroxide or exposure of the airways to house dust mite leads to the release of DAMPs: uric acid, ATP, and IL-1. Exposure to DAMPs or PAMPs present in allergens or added to harmless allergens, such as the experimental allergen ovalbumin, induces several immune responses, including cellular influx and activation. Cellular influx can be analyzed by flow cytometry. Likewise, cellular activation can be assessed by measuring increased expression and release of chemokines and cytokines. These inflammatory mediators can be analyzed by ELISA or confocal microscopy. Here, we describe the protocols for these assessments and a protocol that takes advantage of bone marrow chimeric mice to further elucidate mechanism.


Sulfadoxine/pyrimethamine alone or with amodiaquine or artesunate for treatment of uncomplicated malaria: a longitudinal randomised trial.
New antimalarial treatments are urgently needed in sub-Saharan Africa. Improved therapies should decrease failure rates in the short term, but their effect on incidence of subsequent episodes of malaria is little studied. We aimed to compare the short-term and long-term effectiveness of three antimalarial regimens in children from Kampala, Uganda. We randomly allocated healthy children aged 6 months to 5 years to receive 25 mg/kg sulfadoxine and 1.25 mg/kg pyrimethamine plus either placebo, 25 mg/kg amodiaquine, or 12 mg/kg artesunate. Participants were followed up for 1 year and received the same preassigned treatment for every new episode of uncomplicated malaria diagnosed during follow-up. Recrudescent and new infections were distinguished by comparison of polymorphisms in merozoite surface protein 2 (MSP2). Our primary endpoint was the total number of treatments for malaria per time at risk. Analyses were done per protocol. 183 (61%) of 316 participants were diagnosed with at least one episode of uncomplicated malaria. A total of 577 episodes of uncomplicated Plasmodium falciparum malaria were treated with study drugs; all regimens were safe and well tolerated. Clinical treatment failure after 14 days was significantly more frequent in the sulfadoxine/pyrimethamine group (38 of 215, 18%) compared with either the sulfadoxine/pyrimethamine plus amodiaquine group (two of 164, 1%; p<0.0001) or sulfadoxine/pyrimethamine plus artesunate group (one of 198, 1%; p<0.0001). After 28 and 42 days, patients in the sulfadoxine/pyrimethamine plus amodiaquine group were significantly less likely to develop malaria than were those in the other groups. Overall, sulfadoxine/pyrimethamine plus amodiaquine reduced the rate of subsequent treatments for malaria by 54% (95% CI 36-66, p<0.0001) compared with sulfadoxine/ pyrimethamine alone and by 37% (12-54, p=0.007) compared with sulfadoxine/pyrimethamine plus artesunate. Sulfadoxine/pyrimethamine plus amodiaquine could be used as an inexpensive regimen to decrease the rate of subsequent episodes of malaria.


Clinical Presentations of Brucellosis Over a Four-Year Period at Sultan Qaboos University Hospital and Armed Forces Hospital, Muscat, Oman.
Brucellosis is a highly contagious zoonotic disease which can have serious health implications for affected humans and livestock. This study aimed to evaluate the clinical presentation, geographical distribution and risk factors of brucellosis cases admitted over a four-year period to two hospitals in Muscat, Oman. This observational study was conducted from January 2015 to December 2018 at the Sultan Qaboos University Hospital and Armed Forces Hospital in Muscat. All patients with probable or definitive diagnoses of brucellosis according to the diagnostic criteria of the World Health Organization were included. Relevant data were gathered from the patients' medical records, including results from standard agglutination tests, Brucella enzyme-linked immunosorbent assays, bacterial blood or tissue/aspirate cultures and Brucella polymerase chain reaction tests. A total of 64 patients were diagnosed with brucellosis over the study period. The median age was 31.5 years and 73.4% were male. The majority (95.2%) presented with fever, followed by weight loss (51%), transaminitis (48.4%), peripheral arthritis/arthralgia (15.9%) and back pain (spondylodiscitis/sacroiliitis; 23.4%). Overall, 75.5% reported having consumed raw dairy products, while only 25.9% gave a positive history of animal contact. Patients with brucellosis presented with a wide range of clinical features, the most predominant of which was fever. The majority of patients were residents of or had recently visited Salalah and had consumed raw dairy products. These findings highlight the need for healthcare practitioners to maintain a high index of suspicion for this diagnosis. Moreover, further regulatory measures are necessary to oversee the sale of raw/unpasteurised dairy products.


SLX4 gains weight with SUMO in genome maintenance.
Replication stress has emerged as a key driver of oncogenesis but also represents an Achilles' heel of cancer cells. Newly reported SUMO binding and SUMO ligase functions of the DNA repair protein SLX4 that influence the outcome of replication stress open new avenues for investigating the roles played by SLX4 in tumorigenesis.


Field evaluation of baited traps for surveillance of Aedes japonicus japonicus in Switzerland.
The efficacy of Centers for Disease Control (CDC) miniature light traps and ovitraps was tested in the outskirts of the city of Zurich in Switzerland for their use in the surveillance of Aedes (Hulecoeteomyia) japonicus japonicus (Theobald) (Diptera: Culicidae), the invasive Asian bush mosquito. Sets of single CDC traps were run overnight (n = 18) in three different environments (forest, suburban and urban) in 3 × 3 Latin square experimental designs. Traps were baited with: (a) carbon dioxide (CO2 ); (b) CO2 plus light, or (c) CO2 plus lure blend [Combi FRC 3003 (iGu® )]. At the same locations, mosquito eggs were collected weekly using standard ovitraps baited with different infusions (oak, hay or tap water) and equipped with different oviposition substrates (a block of extruded polystyrene, a germination paper strip or a wooden stick). Data were analysed using Poisson and negative binomial general linear models. The use of light (P < 0.001) or lure (P < 0.001) significantly increased the attractiveness of CDC traps baited with CO2 . Oak and hay infusions did not increase the attractiveness of ovitraps compared with standing tap water (P > 0.05), and extruded polystyrene blocks were preferred as an oviposition substrate over wooden sticks (P < 0.05) and seed germination paper (P < 0.05). Carbon dioxide-baited CDC miniature light traps complemented with light or iGu® lure and ovitraps containing standing tap water and polystyrene oviposition blocks can be considered as efficient and simple tools for use in Ae. j. japonicus surveillance programmes.


Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy.
Cystic fibrosis (CF) is a lethal inherited disorder affecting about 1 in 2,000 Caucasians. The major cause of morbidity is permanent lung damage resulting from ion transport abnormalities in airway epithelia that lead to mucus accumulation and bacterial colonization. CF is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene that encodes a cyclic-AMP-regulated chloride channel. Cyclic-AMP-regulated chloride conductances are altered in airway epithelia from CF patients, suggesting that the functional expression of CFTR in the airways of CF patients may be a strategy for treatment. Transgenic mice with a disrupted cftr gene are appropriate for testing gene therapy protocols. Here we report the use of liposomes to deliver a CFTR expression plasmid to epithelia of the airway and to alveoli deep in the lung, leading to the correction of the ion conductance defects found in the trachea of transgenic (cf/cf) mice. These studies illustrate the feasibility of gene therapy for the pulmonary aspects of CF in humans.


Relative effectiveness of insulin pump treatment over multiple daily injections and structured education during flexible intensive insulin treatment for type 1 diabetes: cluster randomised trial (REPOSE).
Objective To compare the effectiveness of insulin pumps with multiple daily injections for adults with type 1 diabetes, with both groups receiving equivalent training in flexible insulin treatment.Design Pragmatic, multicentre, open label, parallel group, cluster randomised controlled trial (Relative Effectiveness of Pumps Over MDI and Structured Education (REPOSE) trial).Setting Eight secondary care centres in England and Scotland.Participants Adults with type 1 diabetes who were willing to undertake intensive insulin treatment, with no preference for pumps or multiple daily injections. Participants were allocated a place on established group training courses that taught flexible intensive insulin treatment ("dose adjustment for normal eating," DAFNE). The course groups (the clusters) were then randomly allocated in pairs to either pump or multiple daily injections.Interventions Participants attended training in flexible insulin treatment (using insulin analogues) structured around the use of pump or injections, followed for two years.Main outcome measures The primary outcomes were a change in glycated haemoglobin (HbA1c) values (%) at two years in participants with baseline HbA1c value of ≥7.5% (58 mmol/mol), and the proportion of participants achieving an HbA1c value of <7.5%. Secondary outcomes included body weight, insulin dose, and episodes of moderate and severe hypoglycaemia. Ancillary outcomes included quality of life and treatment satisfaction.Results 317 participants (46 courses) were randomised (156 pump and 161 injections). 267 attended courses and 260 were included in the intention to treat analysis, of which 235 (119 pump and 116 injection) had baseline HbA1c values of ≥7.5%. Glycaemic control and rates of severe hypoglycaemia improved in both groups. The mean change in HbA1c at two years was -0.85% with pump treatment and -0.42% with multiple daily injections. Adjusting for course, centre, age, sex, and accounting for missing values, the difference was -0.24% (-2.7 mmol/mol) in favour of pump users (95% confidence interval -0.53 to 0.05, P=0.10). Most psychosocial measures showed no difference, but pump users showed greater improvement in treatment satisfaction and some quality of life domains (dietary freedom and daily hassle) at 12 and 24 months.Conclusions Both groups showed clinically relevant and long lasting decreases in HbA1c, rates of severe hypoglycaemia, and improved psychological measures, although few participants achieved glucose levels currently recommended by national and international guidelines. Adding pump treatment to structured training in flexible intensive insulin treatment did not substantially enhance educational benefits on glycaemic control, hypoglycaemia, or psychosocial outcomes in adults with type 1 diabetes. These results do not support a policy of providing insulin pumps to adults with poor glycaemic control until the effects of training on participants' level of engagement in intensive self management have been determined.Trial registration Current Controlled Trials ISRCTN61215213.


Silent stroke in a case of beta-thalassemia major associated with chronic renal failure and diabetes mellitus.
Severe anemia, growth retardation, diabetes mellitus, cardiac disorders, and, infrequently, stroke are well-known complications of thalassemia major. We report a girl, age 7 years, 2 months, with beta-thalassemia major associated with chronic renal failure, diabetes mellitus, and cardiomyopathy in whom a silent stroke was noted during follow-up. She was diagnosed with thalassemia major at age 6 months, chronic renal failure at age 3 years, 3 months, and diabetes mellitus and cardiomyopathy at age 7 years. Although cranial computed tomography was found to be normal at the age of 3 years, 3 months, magnetic resonance imaging showed cerebral infarct in the right frontal region at 7 years, 2 months. A thrombophilic panel revealed increased factor VIII and decreased protein C concentrations. She died from disseminated intravascular coagulation at age 7 years, 9 months. We did not record any clinical findings of stroke during her follow-up. We think that diabetes mellitus, dilated cardiomyopathy, and increased factor VIII and decreased protein C concentrations led to the occurrence of cerebral infarct. In conclusion, we emphasize that children with thalassemia major should be monitored closely for stroke. We also suggest that stroke can show a silent progression in severely affected children, as in our case.


Implementing the 2005 American Heart Association guidelines, including use of the impedance threshold device, improves hospital discharge rate after in-hospital cardiac arrest.
To determine the impact of the 2005 American Heart Association cardiopulmonary resuscitation (CPR) guidelines, including use of an impedance threshold device (ITD), on survival after in-hospital cardiac arrest. Two community hospitals that tracked outcomes after in-hospital cardiac arrest pooled and compared their hospital discharge rate before and after implementing the 2005 American Heart Association CPR guidelines (including ITD) in standardized protocols. In CPR we used the proper ventilation rate, allowed full chest-wall recoil, conducted continuous CPR following intubation, and used an ITD. We compared historical control data from a 12-month period at St Cloud Hospital, St Cloud, Minnesota, to data from a subsequent 18-month intervention phase. We compared historical control data from a 12-month period at St Dominic Hospital, Jackson, Mississippi to a subsequent 12-month intervention phase. 507 patients received CPR during the study period. Patient age and sex were similar in the control and intervention groups. The combined hospital discharge rate for patients with an in-hospital cardiac arrest was 17.5% in the control group (n=246 patients), which is similar to the national average, versus 28% in the intervention group (n=261 patients) (P=.006, odds ratio 1.83, 95% CI 1.17-2.88). The greatest benefit of the intervention was in patients with an initial rhythm of pulseless electrical activity: 14.4% versus 29.7% (P=.014, odds ratio 2.50, 95% CI 1.15, 5.58). Neurological function (as measured with the Cerebral Performance Category scale) in survivors at hospital discharge was similar between the groups. Implementation of improved ways to increase circulation during CPR increased the in-hospital discharge rate by 60%, compared to historical controls in 2 community hospitals. These data demonstrate that immediate care with improved means to circulate blood during CPR significantly reduces hospital mortality from inhospital sudden cardiac arrest.


Purification and characterization of novel fibrinolytic proteases as potential antithrombotic agents from earthworm Perionyx excavatus.
Six protease fractions, namely FI, FII, FIII-1, FIII-2, FIII-3 and FIV, were isolated from Perionyx excavatus earthworm biomass by acetone precipitation, followed by serial chromatography using anion exchange, hydrophobic interaction and size exclusion chromatography. All fractions exhibited strong hydrolytic activity towards casein. The activity of six fractions towards fibrin, determined by fibrin plate assay, ranged from 44 to 831 plasmin unit.mg-1 and ranked as FIII-3 > FIII-2 > FI > FIII-1 > FIV > FII. Casein degradation was optimal at pH 7 and 11, and at 45-60°C. All fractions were considerably stable at high temperature and wide pH range. They were completely inhibited by phenylmethylsulfonyl fluoride (PMSF). The molecular weights (MW) and isoelectric points (pI) determined by 2D-electrophoresis were 27.5-34.5 kDa, and 4.3-5.2, respectively. Tandem mass spectrometry (MS) analysis was used to deduce the amino acid sequences of some peptides from FIII-1 and FIII-2. The sequences shared 16.9% and 13.2% similarity, respectively, with the fibrinolytic enzymes from two related earthworm species, Lumbricus rubellus and Eisenia fetida. The P. excavatus proteases were classified as serine proteases. They could perform rapid hydrolysis on both coagulated fibrous fibrin and soluble fibrinogen monomers without the presence of activators such as tPA or urokinase.


Looking like a leader-facial shape predicts perceived height and leadership ability.
Judgments of leadership ability from face images predict the outcomes of actual political elections and are correlated with leadership success in the corporate world. The specific facial cues that people use to judge leadership remain unclear, however. Physical height is also associated with political and organizational success, raising the possibility that facial cues of height contribute to leadership perceptions. Consequently, we assessed whether cues to height exist in the face and, if so, whether they are associated with perception of leadership ability. We found that facial cues to perceived height had a strong relationship with perceived leadership ability. Furthermore, when allowed to manually manipulate faces, participants increased facial cues associated with perceived height in order to maximize leadership perception. A morphometric analysis of face shape revealed that structural facial masculinity was not responsible for the relationship between perceived height and perceived leadership ability. Given the prominence of facial appearance in making social judgments, facial cues to perceived height may have a significant influence on leadership selection.


Stromal cell-derived factor 1 as a biomarker of heart failure and mortality risk.
CXCL12 encodes stromal cell-derived factor 1α (SDF-1), which binds to the receptor encoded by CXCR4. Variation at the CXCL12 locus is associated with coronary artery disease and endothelial progenitor cell numbers, whereas variation at the CXCR4 locus is associated with leukocyte telomere length, which has been shown to be associated with coronary artery disease. Therefore, we examined the relationships of plasma SDF-1 levels to cardiovascular disease (CVD)-related outcomes, risk factors, leukocyte telomere length, and endothelial progenitor cells. SDF-1 was measured in 3359 Framingham Heart Study participants. We used Cox regression to examine relationships of SDF-1 to new-onset CVD, myocardial infarction, heart failure, and all-cause mortality; we used linear regression to evaluate associations of SDF-1 with risk factors, leukocyte telomere length, and CD34+ cell phenotypes. In multivariable models, higher SDF-1 levels were associated with older age, lower levels of high-density lipoprotein-cholesterol and cigarette smoking. Higher SDF-1 levels were associated with lower CD34+ cell frequency (P=0.02) but not with leukocyte telomere length. During follow-up (median, 9.3 years), there were 263 new-onset CVD events, 160 myocardial infarctions, 200 heart failure events, and 385 deaths. After adjusting for clinical risk factors, SDF-1 levels were associated with heart failure (P=0.04) and all-cause mortality (P=0.003) but not with CVD (P=0.39) or myocardial infarction (P=0.10). The association of SDF-1 levels with myocardial infarction was attenuated after adjustment for high-density lipoprotein-cholesterol. After adjusting for traditional CVD risk factors, SDF-1 is associated with heart failure and all-cause mortality risk. Additional studies are needed to determine whether measurement of SDF-1 levels has clinical use.


Papillary renal cell carcinoma: quantitation of chromosomes 7 and 17 by FISH, analysis of chromosome 3p for LOH, and DNA ploidy.
Papillary renal cell carcinoma (papillary RCC) is an uncommon histologic variant of RCC with distinct gross, microscopic, and immunohistochemical features. Recent karyotypic analyses suggest that papillary RCC differs from other types of RCC at the genetic level as well. Whereas nonpapillary (clear cell, granular cell) RCC is characterized by deletions in chromosome 3p, papillary tumors reportedly exhibit a pattern of chromosomal trisomies, typically including chromosomes 7 and 17. To further examine the relationship between overrepresentation of these chromosomes and papillary histology, archival material from 36 papillary tumors was subjected to fluorescence in situ hybridization (FISH) analysis using alpha-satellite repeat probes specific to 7 and 17. Excess signals for chromosome 17 were detected in 22 of 28 (78%) low-grade papillary tumors (Fuhrman nuclear grades 1 and 2), and in seven of eight (87%) high-grade tumors (grades 3 and 4). Correlation of chromosome 17 FISH signals with karyotypes performed on two low-grade and three high-grade tumors was excellent. Among the cases without evidence of excess chromosome 17 were three unusual papillary tumors with sclerotic and hyalinized fibrovascular cores. In two cases, comparison was made of FISH signals from multiple, separate gross nodules of tumor; concordance for trisomic 17 signals was observed in one case, but not in the other. Chromosome 7 signals were overrepresented in all seven papillary tumors examined. DNA ploidy was determined in 19 of the 36 tumors; a relationship between DNA ploidy and polysomy 7 or 17 was not apparent. To examine the possible role of chromosome 3p deletions in the development of papillary RCC, 11 cases were studied for loss of heterozygosity (LOH) at one or more loci in the region of 3p13-21. Only three of the 11 cases had LOH at these loci. The findings are discussed with respect to the development and progression of papillary RCC.


Telephone interviews can be used to collect follow-up data subsequent to no response to postal questionnaires in clinical trials.
Follow-up data were collected using postal questionnaires and if participants did not respond, then data was collected using telephone interviews. The objectives of this study were to examine, for the two methods, how respondents differed in characteristics and whether the observed treatment difference varied. A large clinical trial of lower back pain. About 60% (98/163) of the nonresponders to postal questionnaire provided data by telephone, which increased the overall response rate by 14% (from 71% to 85%). A consistent treatment difference was found across the methods for the outcome measures at 12 months, implying that the observed treatment effect had not been modified. There were some differences between the participants: responders of postal questionnaire were older, likely to be female, white (ethnic origin), not working, with less disability of back pain, compared with those who responded by a telephone interview. At 12 months, there was greater improvement in back pain, disability, and general health for those who responded by postal questionnaires. Researchers should consider the use of more than one method of collecting data as this increases response rate, participant representativeness, and enhances precision of effect estimates.


Porcine models for the metabolic syndrome, digestive and bone disorders: a general overview.
The aim of this review article is to provide an overview of the role of pigs as a biomedical model for humans. The usefulness and limitations of porcine models have been discussed in terms of metabolic, cardiovascular, digestive and bone diseases in humans. Domestic pigs and minipigs are the main categories of pigs used as biomedical models. One drawback of minipigs is that they are in short supply and expensive compared with domestic pigs, which in contrast cost more to house, feed and medicate. Different porcine breeds show different responses to the induction of specific diseases. For example, ossabaw minipigs provide a better model than Yucatan for the metabolic syndrome as they exhibit obesity, insulin resistance and hypertension, all of which are absent in the Yucatan. Similar metabolic/physiological differences exist between domestic breeds (e.g. Meishan v. Pietrain). The modern commercial (e.g. Large White) domestic pig has been the preferred model for developmental programming due to the 2- to 3-fold variation in body weight among littermates providing a natural form of foetal growth retardation not observed in ancient (e.g. Meishan) domestic breeds. Pigs have been increasingly used to study chronic ischaemia, therapeutic angiogenesis, hypertrophic cardiomyopathy and abdominal aortic aneurysm as their coronary anatomy and physiology are similar to humans. Type 1 and II diabetes can be induced in swine using dietary regimes and/or administration of streptozotocin. Pigs are a good and extensively used model for specific nutritional studies as their protein and lipid metabolism is comparable with humans, although pigs are not as sensitive to protein restriction as rodents. Neonatal and weanling pigs have been used to examine the pathophysiology and prevention/treatment of microbial-associated diseases and immune system disorders. A porcine model mimicking various degrees of prematurity in infants receiving total parenteral nutrition has been established to investigate gut development, amino acid metabolism and non-alcoholic fatty liver disease. Endoscopic therapeutic methods for upper gastrointestinal tract bleeding are being developed. Bone remodelling cycle in pigs is histologically more similar to humans than that of rats or mice, and is used to examine the relationship between menopause and osteoporosis. Work has also been conducted on dental implants in pigs to consider loading; however with caution as porcine bone remodels slightly faster than human bone. We conclude that pigs are a valuable translational model to bridge the gap between classical rodent models and humans in developing new therapies to aid human health.


Effect of trans fatty acid intake on abdominal and liver fat deposition and blood lipids: a randomized trial in overweight postmenopausal women.
Intake of industrially produced trans fatty acids (TFAs) is, according to observational studies, associated with an increased risk of cardiovascular disease, but the causal mechanisms have not been fully elucidated. Besides inducing dyslipidemia, TFA intake is suspected to promote abdominal and liver fat deposition. We examined the effect of a high intake of TFA as part of an isocaloric diet on whole-body, abdominal and hepatic fat deposition, and blood lipids in postmenopausal women. In a 16-week double-blind parallel intervention study, 52 healthy overweight postmenopausal women were randomized to receive either partially hydrogenated soybean oil providing 15.7 g day(-1) of TFA or a control oil with mainly oleic and palmitic acid. Before and after the intervention, body composition was assessed by dual-energy X-ray absorptiometry, abdominal fat by magnetic resonance (MR) imaging, and liver fat by (1)H MR spectroscopy. Compared with the control fat, TFA intake decreased plasma high-density lipoprotein (HDL)-cholesterol by 10%, increased low-density lipoprotein (LDL)-cholesterol by 18% and resulted in an increased LDL/HDL-cholesterol ratio (baseline adjusted mean (95% CI) difference between diet groups 0.41 (0.22; 0.60); P<0.001). TFA tended to increase the body fat (0.46 (-0.20; 1.17) kg; P=0.16) and waist circumference (1.1 (-0.1; 2.4) cm; P=0.08) more than the control fat, whereas neither abdominal nor liver fat deposition was affected by TFA. The adverse effect of dietary TFA on cardiovascular disease risk involves induction of dyslipidemia, and perhaps body fat, whereas weight gain-independent accumulation of ectopic fat could not be identified as a contributory factor during short-term intake.


Phase I and pharmacokinetic studies of CYT-6091, a novel PEGylated colloidal gold-rhTNF nanomedicine.
A novel nanomedicine, CYT-6091, constructed by simultaneously binding recombinant human tumor necrosis factor alpha (rhTNF) and thiolyated polyethylene glycol to the surface of 27-nm colloidal gold particles, was tested in a phase I dose escalation clinical trial in advanced stage cancer patients. CYT-6091, whose dosing was based on the amount of rhTNF in the nanomedicine, was injected intravenously, and 1 cycle of treatment consisted of 2 treatments administered 14 days apart. Doses from 50 μg/m(2) to 600 μg/m(2) were well tolerated, and no maximum tolerated dose (MTD) was reached, as the highest dose exceeded the target dosage of 1-mg rhTNF per treatment, exceeding the previous MTD for native rhTNF by 3-fold. The first 2 patients on the study, each receiving 50 μg/m(2), did not receive any prophylactic antipyretics or H2 blockade. A predicted, yet controllable fever occurred in these patients, so all subsequently treated patients received prophylactic antipyretics and H2 blockers. However, even at the highest dose rhTNF's dose-limiting toxic effect of hypotension was not seen. Using electron microscopy to visualize nanoparticles of gold in patient biopsies of tumor and healthy tissue showed that patient biopsies taken 24 hours after treatment had nanoparticles of gold in tumor tissue. These data indicate that rhTNF formulated as CYT-6091 may be administered systemically at doses of rhTNF that were previously shown to be toxic and that CYT-6091 may target to tumors. Future clinical studies will focus on combining CYT-6091 with approved chemotherapies for the systemic treatment of nonresectable cancers.


High efficiency human memory B cell assay and its application to studying Plasmodium falciparum-specific memory B cells in natural infections.
Memory B cells (MBCs) are a key component of long term humoral immunity to many human infectious diseases. Despite their importance, we know little about the generation or maintenance of antigen-(Ag)-specific MBCs in humans in response to infection. A frequently employed method for quantifying Ag-specific MBCs in human peripheral blood (Crotty et al., 2004) relies on the ability of MBCs but not naïve B cells to differentiate into antibody secreting cells (ASCs) in response to polyclonal activators and Toll-like receptor agonists in vitro and the measurement of Ag-specific ASCs by ELISPOT assays. Here we report on studies to optimize the efficiency of this ELISPOT-based assay and to apply this assay to the detection of Plasmodium falciparum (Pf)-specific MBCs in adults living in a malaria endemic area where immunity to Pf is acquired through natural infection. We show that the addition of IL-10 to in vitro cultures of human peripheral blood mononuclear cells increased the efficiency of the assay from 10% to over 90% without increasing the ASC burst size and without any substantial increase in background from naïve B cells or plasma cells (PCs). Using this assay we were able to quantify the frequency of Pf-specific MBCs in peripheral blood of adults living in a malaria endemic area. Thus, this highly efficient assay appears to be well suited to field studies of the generation and maintenance of MBCs where the volumes of blood obtainable are often limiting.


Mutations affecting RNA polymerase I-stimulated exchange and rDNA recombination in yeast.
HOT1 is a cis-acting recombination-stimulatory sequence isolated from the rDNA repeat unit of yeast. The ability of HOT1 to stimulate mitotic exchange appears to depend on its ability to promote high levels of RNA polymerase I transcription. A qualitative colony color sectoring assay was developed to screen for trans-acting mutations that alter the activity of HOT1. Both hypo-recombination and hyper-recombination mutants were isolated. Genetic analysis of seven HOT1 recombination mutants (hrm) that decrease HOT1 activity shows that they behave as recessive nuclear mutations and belong to five linkage groups. Three of these mutations, hrm1, hrm2, and hrm3, also decrease rDNA exchange but do not alter recombination in the absence of HOT1. Another mutation, hrm4, decreases HOT1-stimulated recombination but does not affect rDNA recombination or exchange in the absence of HOT1. Two new alleles of RAD52 were also isolated using this screen. With regard to HOT1 activity, rad52 is epistatic to all four hrm mutations indicating that the products of the HRM genes and of RAD52 mediate steps in the same recombination pathway. Finding mutations that decrease both the activity of HOT1 and exchange in the rDNA supports the hypothesis that HOT1 plays a role in rDNA recombination.


HLA-DR alpha gene regulation in immortalized human thyroid cancer cells.
The induction of HLA class II antigens on human thyroid epithelial cells has been shown to be an intimate part of the pathology of autoimmune thyroid disease and may also be relevant to the natural history of thyroid neoplasia since thyroid cancer cells may show spontaneous HLA class II antigen expression. We have, therefore, analyzed the regulation of HLA-DR-alpha-specific mRNA transcripts, as a model for HLA class II antigen induction, in three established human thyroid cancer cell lines (papillary thyroid cell line NPA, and follicular thyroid cell lines RO-82-W1 and MRO-87-1). Each of the lines expressed 1.4 kb HLA-DR-alpha-specific mRNA transcripts, either constitutively or after cytokine induction, but showed markedly different regulatory characteristics. For example, the induction of HLA-DR alpha mRNA in response to recombinant human interferon-gamma (IFN-gamma) was inversely proportional to the degree of constitutive DR alpha mRNA expression; when there was constitutive absence there was a greater degree of induction. Similarly, the half-lives of the HLA-DR alpha mRNA transcripts varied from 80 to 420 min with the longest degradation time occurring in those cells lacking constitutive expression of HLA-DR alpha mRNA. These data indicated that the degradation rate was not a determinant of their constitutive expression. We also sequenced the 5' promoter region of the HLA-DR alpha gene (nucleotides -88 to -277 with respect to the translation start site) in each of the three cell lines. Sequence analyses revealed identity to the previously published normal human genomic sequence. Taken together, therefore, these data indicate that transactivating factors, rather than changes in mRNA degradation or promoter abnormalities, are the likely causes of variation in constitutive and cytokine-induced HLA-DR alpha gene expression in human thyroid cancer cells.


Common tolerance mechanisms, but distinct cross-reactivities associated with gp41 and lipids, limit production of HIV-1 broad neutralizing antibodies 2F5 and 4E10.
Developing an HIV-1 vaccine has been hampered by the inability of immunogens to induce broadly neutralizing Abs (BnAbs) that protect against infection. Previously, we used knockin (KI) mice expressing a prototypical gp41-specific BnAb, 2F5, to demonstrate that immunological tolerance triggered by self-reactivity of the 2F5 H chain impedes BnAb induction. In this study, we generate KI models expressing H chains from two other HIV-1 Abs, 4E10 (another self-/polyreactive, anti-gp41 BnAb) and 48d (an anti-CD4 inducible, nonpolyreactive Ab), and find a similar developmental blockade consistent with central B cell deletion in 4E10, but not in 48d VH KI mice. Furthermore, in KI strains expressing the complete 2F5 and 4E10 Abs as BCRs, we find that residual splenic B cells arrest at distinct developmental stages, yet exhibit uniformly low BCR densities, elevated basal activation, and profoundly muted responses to BCR ligation and, when captured as hybridoma mAb lines, maintain their dual (gp41/lipid) affinities and capacities to neutralize HIV-1, establishing a key role for anergy in suppressing residual 2F5- or 4E10-expressing B cells. Importantly, serum IgGs from naive 2F5 and 4E10 KI strains selectively eliminate gp41 and lipid binding, respectively, suggesting B cells expressing 2F5 or 4E10 as BCRs exhibit specificity for a distinct spectrum of host Ags, including selective interactions by 2F5 BCR(+) B cells (i.e., and not 4E10 BCR(+) B cells) with those mimicked by its gp41 neutralization epitope.


Comparison of X-ray crystal structures of a tetradecamer sequence d(CCCGGGTACCCGGG)2 at 1.7 Å resolution.
We present here a comparison of three different X-ray crystal structures of DNA tetradecamer sequence d(CCCGGGTACCCGGG)2 all at about 1.7 Å resolution. The sequence was designed as an attempt to form a DNA four-way junction with A-type helical arms. However, in the presence of zinc, magnesium, and in the absence of any metal ion, it does not take up the junction structure, but forms an A-type double helix. This allowed us to study possible conformational changes in the double helix due to the presence of metal ions. Upon addition of the zinc ion, there is a change in the space group from P41212 to P41. The overall conformation of the duplex remains the same. There are small changes in the interaction of the metal ions with the DNA. In the zinc-bound structure, there are two zinc ions that show direct interaction with the N7 atoms of terminal G13 bases at either end of the molecule. There are small changes in the interhelical contacts. The consequence of these differences is to break some of the symmetry and change the space group.


A novel transcriptional factor important for pathogenesis and ascosporogenesis in Fusarium graminearum.
Fusarium head blight or scab caused by Fusarium graminearum is an important disease of wheat and barley. The pathogen not only causes severe yield losses but also contaminates infested grains with mycotoxins. In a previous study, we identified several pathogenicity mutants by random insertional mutagenesis. One of these mutants was disrupted in the ZIF1 gene, which encodes a b-ZIP transcription factor unique to filamentous ascomycetes. The Δzif1 mutant generated by gene replacement was significantly reduced in deoxynivalenol (DON) production and virulence on flowering wheat heads. It was defective in spreading from inoculated florets to the rachis and other spikelets. Deletion of the ZIF1 ortholog MoZIF1 in the rice blast fungus also caused reductions in virulence and in invasive growth. In addition, the Δzif1 mutant is defective in sexual reproduction. Although it had normal male fertility, when selfed or mated as the female in outcrosess, the Δzif1 mutant produced small, pigmented perithecia that were sterile (lack of asci and ascospores), suggesting a female-specific role for ZIF1 during fertilization or ascus development. Similar female-specific defects in sexual reproduction were observed in the ΔMozif1 mutant. When mated as the female, the ΔMozif1 perithecia failed to develop long necks and asci or ascospores. The ZIF1 gene is well conserved in filamentous ascomycetes, particularly in the b-ZIP domain, which is essential for its function. Expression of ZIF1 in Magnaporthe oryzae complemented the defects of the ΔMozif1 mutant. These results indicate that this b-ZIP transcription factor is functionally conserved in these two fungal pathogens for plant infection and sexual reproduction.


Policy statement--health equity and children's rights.
Many children in the United States fail to reach their full health and developmental potential. Disparities in their health and well-being result from the complex interplay of multiple social and environmental determinants that are not adequately addressed by current standards of pediatric practice or public policy. Integrating the principles and practice of child health equity-children's rights, social justice, human capital investment, and health equity ethics-into pediatrics will address the root causes of child health disparities. Promoting the principles and practice of equity-based clinical care, child advocacy, and child- and family-centered public policy will help to ensure that social and environmental determinants contribute positively to the health and well-being of children. The American Academy of Pediatrics and pediatricians can move the national focus from documenting child health disparities to advancing the principles and practice of child health equity and, in so doing, influence the worldwide practice of pediatrics and child health. All pediatricians, including primary care practitioners and medical and surgical subspecialists, can incorporate these principles into their practice of pediatrics and child health. Integration of these principles into competency-based training and board certification will secure their assimilation into all levels of pediatric practice.


Regulatory T cells and IgE expression in duodenal mucosa of Strongyloides stercoralis and human T lymphotropic virus type 1 co-infected patients.
Strongyloides stercoralis is an intestinal nematode unique in its ability to replicate in the human host, allowing ongoing cycles of autoinfection, persisting for decades within the same host. Although usually asymptomatic, overwhelming infections can occur in Strongyloides and HTLV-1 co-infected individuals (SS/HTLV-1). Regulatory T cells (Tregs) are able to blunt specific Th2 responses necessary to control the parasite. We previously reported that peripheral blood Tregs are increased in SS/HTLV-1 and correlate with low Th2 responses. We hypothesized that Tregs are also increased at the site of infection in duodenal mucosa. Paraffin embedded duodenal biopsies were obtained from 10 SS/HTLV-1 patients, 3 controls with non-parasitic chronic duodenitis, and 2 healthy controls. Immunohistochemistry was performed using monoclonal antibodies against human CD3, CD8, IgE and FoxP3. The number of cells were counted using a conventional light microscope. The number of CD3+, CD8+, FoxP3+ and IgE positive cells per 0.35 mm2 was measured using ImagePro Plus software comparing areas adjacent or distant from parasite material. In patients with SS/HTLV-1, T lymphocyte counts and CD8+ cells were lower in areas adjacent to the parasite compared to non-adjacent areas (CD3+: adjacent: 6.5 [Interquartile range (IQR: 2.8-12.3)]; non-adjacent: 24.5 [IQR: 20.9-34.4]; Mann-Whitney p = 0.0003; CD8+: adjacent: 4.5 [IQR: 2.3-11.8]; non-adjacent: 21 [IQR: 15.3-42.9]; Mann-Whitney p = 0.0011). Tregs cells in the intestines (FoxP3+ expressing cells) were increased in patients with SS/HTLV-1 compared with patients with chronic duodenitis (SS/HTLV-1: 1.5 [IQR: 0.7-2.3]; duodenitis controls: 0 [range 0-0.7]; healthy controls: 0; Mann-Whitney p = 0.034). There was also a trend towards fewer eosinophils adjacent to the parasites. Among SS/HTLV-1 patients the number of IgE expressing cells was increased for in areas not adjacent to the parasite compared to non-adjacent areas (ANOVA, p = 0.001). Our data shows increased Treg cell numbers localized adjacent to the parasites in the duodenum SS/HTLV-1 patients. In addition, other T lymphocytes and IgE expressing cells were decreased adjacent to the parasites, suggesting an important role for Tregs in down-regulating local parasite effector responses.


Single-ion heat capacities, C(p)(298)ion, of solids: with a novel route to heat-capacity estimation of complex anions.
Single-ion heat capacities, C(p)(298)(ion), are additive values for the estimation of room-temperature (298 K) heat capacities of ionic solids. They may be used for inferring the heat capacities of ionic solids for which values are unavailable and for checking reported values, thus complementing our independent method of estimation from formula unit volumes (termed volume-based thermodynamics, VBT). Analysis of the reported heat-capacity data presented here provides a new self-consistent set of heat capacities for both cations and anions that is compatible (and thus may be combined) with an extensive set developed by Spencer. The addition of a large range of silicate species permits the estimation of the heat capacities of many silicate minerals. The single-ion heat capacities of individual silicate anions are observed to be strictly proportional to the total number of atoms (Si plus O), n, contained within the silicate anion complex itself (e.g., for the anion Si(2)O(7)(2-), n = 9, for SiO(4)(2-), n = 5), C(p)(silicate anion)/J K(-1) mol(-1) = 13.8n, in a new rule that is an extension of the Neumann-Kopp relationship. The same linear relationship applies to other homologous anion series (for example, oxygenated heavy-metal anion complexes such as niobates, bismuthates, and tantalates), although with a different proportionality constant. A similar proportionality, C(p)(complex anion)/J K(-1) mol(-1) ≈ 17.5n, which may be regarded as a convenient "rule of thumb", also applies, although less strictly, to complex anions in general. The proportionality constants reflect the rigidity of the complex anion, being always less than the Dulong-Petit value of 25 J K(-1) mol(-1). An emergent feature of our VBT and single-ion approaches to an estimation of the thermodynamic properties is the identification of anomalies in measured values, as is illustrated in this paper.


Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor.
Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/ threonine kinase Akt, which then phosphorylates and inactivates components of the apoptotic machinery, including BAD and Caspase 9. In this study, we demonstrate that Akt also regulates the activity of FKHRL1, a member of the Forkhead family of transcription factors. In the presence of survival factors, Akt phosphorylates FKHRL1, leading to FKHRL1's association with 14-3-3 proteins and FKHRL1's retention in the cytoplasm. Survival factor withdrawal leads to FKHRL1 dephosphorylation, nuclear translocation, and target gene activation. Within the nucleus, FKHRL1 triggers apoptosis most likely by inducing the expression of genes that are critical for cell death, such as the Fas ligand gene.


Visualization of Lymphatic Vessels Using Photoacoustic Imaging.
Lymphedema occurs when interstitial fluid and fibroadipose tissues accumulate abnormally because of decreased drainage of lymphatic fluid as a result of injury, infection, or congenital abnormalities of the lymphatic system drainage pathway. An accurate anatomical map of the lymphatic vasculature is needed not only for understanding the pathophysiology of lymphedema but also for surgical planning. However, because of their limited spatial resolution, no imaging modalities are currently able to noninvasively provide a clear visualization of the lymphatic vessels. Photoacoustic imaging is an emerging medical imaging technique that provides unique scalability of optical resolution and acoustic depth of penetration. Moreover, light-absorbing biomolecules, including oxy- and deoxyhemoglobin, lipids, water, and melanin, can be imaged. Using exogenous contrast agents that are taken up by lymphatic vessels, e.g., indocyanine green, photoacoustic lymphangiography, which has a higher spatial resolution than previous imaging modalities, is possible. Using a new prototype of a photoacoustic imaging system with a wide field of view developed by a Japanese research group, high-resolution three-dimensional structural information of the vasculatures was successfully obtained over a large area in both healthy and lymphedematous extremities. Anatomical information on the lymphatic vessels and adjacent veins provided by photoacoustic lymphangiography is helpful for the management of lymphedema. In particular, such knowledge will facilitate the planning of microsurgical lymphaticovenular anastomoses to bypass the excess fluid component by joining with the circulatory system peripherally. Although challenges remain to establish its implementation in clinical practice, photoacoustic lymphangiography may contribute to improved treatments for lymphedema patients in the near future.


Selective substrate-based inhibitors of mammalian dimethylarginine dimethylaminohydrolase.
The enzyme DDAH metabolizes methylarginines that are inhibitors of nitric oxide synthase (NOS). Substrate-based inhibitors of mammalian DDAH have been synthesized, with optimization to give selective inhibition of DDAH with no significant direct effect on NOSs. These are the first examples of reversible DDAH inhibitors with significant activity and selectivity. In vivo administration increases plasma ADMA levels, giving proof of concept that these inhibitors can be used to probe the physiological effects of DDAH inhibition, with potential for pharmaceutical use of DDAH inhibitors in diseases where excess NO production is implicated.


PDIA, an Iminosugar Compound with a Wide Biofilm Inhibitory Spectrum Covering Both Gram-Positive and Gram-Negative Human Bacterial Pathogens.
Many difficult-to-treat human infections related to catheters and other indwelling devices are caused by bacteria residing in biofilms. One of the key properties of microorganisms residing in a biofilm is decreased susceptibility towards antimicrobial agents. Therefore, many different approaches have been researched to destroy or inhibit biofilm production by bacteria. Different iminosugars (IS) were reported to inhibit biofilm formation in S. mutans, S. aureus, and P. aeruginosa. The aim of this study was to look for a spectrum of the activity in one of these IS. The iminosugar PDIA beta-1-C-propyl-1,4-dideoxy-1,4-imino-L-arabinitol was tested in vitro at the same concentration against 30 different strains of the most important Gram-negative and Gram-positive human pathogens looking for their biofilm production and viability at different time intervals. It appeared that PDIA inhibited biofilm production of Enterobacter spp., P. aeruginosa, Enterococcus spp. and S. aureus in 8 h, and Klebsiella spp., Acinetobacter spp. and S.epidermidis in 24 h. PDIA caused no growth inhibition of the tested bacteria at a concentration of 0.9 mM. Our results indicate a broad-spectrum biofilm inhibitory activity of PDIA. which may be the basis for future application studies that will help in control of the associated device and biofilm-related infections caused by a wide spectrum of the causative agents.


Pediatric Neuromuscular Disorders.
Neuromuscular disorders are pathologies that can severely affect the quality of life as well as longevity of patients. The most common disorders include cerebral palsy and myelodysplasia. The orthopedic manifestations of these disorders can be treated operatively or nonoperatively. Both focus on the prolongation of mobility and preservation of ambulatory capacity for patients.


Obesity and the labor market: A fresh look at the weight penalty.
This paper applies semiparametric regression models to shed light on the relationship between body weight and labor market outcomes in Germany. We find conclusive evidence that these relationships are poorly described by linear or quadratic OLS specifications. Women's wages and employment probabilities do not follow a linear relationship and are highest at a body weight far below the clinical threshold of obesity. This indicates that looks, rather than health, is the driving force behind the adverse labor market outcomes to which overweight women are subject. Further support is lent to this notion by the fact that wage penalties for overweight and obese women are only observable in white-collar occupations. On the other hand, bigger appears to be better in the case of men, for whom employment prospects increase with weight, albeit with diminishing returns. However, underweight men in blue-collar jobs earn lower wages because they lack the muscular strength required in such occupations.


Study on coccidiosis of scavenging indigenous chickens in Central Ethiopia.
An investigation was made into coccidiosis of 190 scavenging indigenous chickens between September 2000 and April 2001 in three selected agroclimatic zones, in central Ethiopia. This was done through clinical, postmortem and microscopic examinations. Data were processed by chi-square and Mantel-Haenzel test. The study indicated that 25.8% (49/190) of the chickens were infected with coccidiosis and found to harbour one to four different species of Eimeria. Of these infected chickens, 30 (15.8%) and 19 (10.0%) were positive for clinical and sub-clinical coccidiosis, respectively. There was a significant altitude difference (chi2 = 14.7, p <0.001) in coccidiosis prevalence: 42.2% in chickens from highland region followed by 21.5% in mid-altitude and 13.1% in low-altitude areas. When quantified, the prevalence of coccidiosis was 2.66 and 4.83 times higher in the high-altitude than in mid-altitude (odds ratio, OR = 2.66, p<0.05) and low-altitude (OR = 4.83, p<0.001) chickens. The pathogenic Eimeria species responsible for clinical coccidiosis were E. necatrix, E. acervulina, E. maxima and E. tenella. With increasing demand for poultry products in developing countries, knowledge of production constraints in traditional management practices could help devise control strategies for constraints on backyard poultry production systems.


Autoantibodies in the sera of breast cancer patients: Antinuclear and anti-double stranded DNA antibodies as example.
Inflammation and cell necrosis are one of the consequences that accompany breast cancer. However, inflammation and cell necrosis are well known to be involved in stimulation of cellular and humeral immunity. The aim of this study is to investigate the immune response to the inflammation that accompanies cancer through measuring plasma concentration of antinuclear antibodies (ANAs) and anti-double stranded deoxyribonucleic acid antibodies (ADSDAs). Thirty-five newly diagnosed breast cancer patients were involved in this study from the Radiation Isotopes Center Khartoum (RICK) compared to 18 age- and sex-matched control subjects. Intravenous blood sample was obtained from each study subject and Enzyme Linked Immuno Sorbent Assay (ELISA) technique was used to determine the concentration of the two antibodies. Regarding the ANA concentration in the patients; the range was 0.7-1.8 IU/ml, mean was 0.96, and the standard deviation (SD) was 0.25; while the range of the concentration in the control subjects was 0.3-0.6 IU/ml, mean was 0.47, and SD was 0.07. However, when the means of patients and controls were compared, the difference was significant (P < 0.000). Concerning the result anti-double stranded DNA (anti-dsDNA), its concentration range in the patients was 2.6-151.9 IU/ml, the mean was 55.2, and SD was 25.6, while in healthy people concentration range was 26.1-97.3 IU/ml, the mean was 50.3, and SD was 16.9. There was no significant change between the patients and controls (P = 0.46). The ANA concentration in the patients was significantly increased, while there was no significant difference between the results of ADSDAs in the patients and the control subjects.


1,2,4-Dithiazole-5-ones and 5-thiones as efficient sulfurizing agents of phosphorus(III) compounds--a kinetic comparative study.
The sulfurization efficiency of 25 3-substituted-1,2,4-dithiazole-5-ones and 5-thiones towards triphenyl phosphite in acetonitrile, DCM, THF and toluene at 25 °C was evaluated. All the 1,2,4-dithiazoles are much better sulfurizing reagents than commercially available agents (PADS, TETD, Beaucage's reagent). The most efficient sulfurizing agents in all solvents are 3-phenoxy (4), 3-phenylthio (5) and 3-ethoxy-1,2,4-dithiazole-5-one (1) whose reactivity is at least two orders of magnitude higher than that of other 1,2,4-dithiazoles. Contrary to a previous report, the sulfurization with 1 does not yield carbonylsulfide and ethyl cyanate as the additional reaction products but unstable ethoxythiocarbonyl isocyanate which has been trapped with 4-methoxyaniline. Similar trapping experiments have proven that the site of attack is at the sulfur adjacent to the C=O group for compounds 4 and 5. The reaction pathway involves rate-limiting initial nucleophilic attack of the phosphorus at sulfur followed by decomposition of the phosphonium intermediate to the corresponding phosphorothioate and isocyanate/isothiocyanate species. The existence of the phosphonium intermediate during sulfurization of triphenyl phosphine with 3-phenyl-1,2,4-dithiazole-5-thione (7a) was proven using kinetic studies. From the Hammett and Brønsted correlations and from other kinetic measurements it was concluded that the transition-state structure is almost apolar for the most reactive 1,2,4-dithiazoles whereas a polar structure resembling a zwitter-ionic intermediate may be more appropriate for the least reactive 1,2,4-dithiazoles. The extent of P–S bond formation and S–S bond cleavage is very similar in all reaction series but it gradually decreases with the reactivity of the 1,2,4-dithiazole derivatives.


Choosing the optimal wall shear parameter for the prediction of plaque location-A patient-specific computational study in human left coronary arteries.
While the correlation of atherosclerotic plaque locations with local wall shear stress magnitude has been evaluated previously by other investigators in both right (RCA) and left coronary arteries (LCA), the relative performance of average wall shear stress (AWSS), average wall shear stress gradient (AWSSG), oscillatory shear index (OSI) and relative residence time (RRT) as indicators of potential atherosclerotic plaque locations has not been studied for the LCA. Here we determine the performance of said wall shear parameters in the LCA for the prediction of plaque development locations and compare these results to those previously found in the RCA. We obtained 30 patient-specific geometries (mean age 67.1 (± 9.2) years, all with stable angina) of the LCA using dual-source computed tomography and virtually removed any plaque present. We then performed computational fluid dynamics simulations to calculate the wall shear parameters. For the 96 total plaques, AWSS had a higher sensitivity for the prediction of plaque locations (86 ± 25%) than AWSSG (65 ± 37%, p<0.05), OSI (67 ± 32%, p<0.01) or RRT (48 ± 38%, p<0.001). RRT had a higher PPV (49 ± 36%) than AWSS (31 ± 20%, p<0.05) or AWSSG (16 ± 12%, p<0.001). Segment 5 of the LCA presented with overall low values for sensitivity and PPV. Parameter performance in the remainder of the LCA was comparable to that in the RCA. AWSS features remarkably high sensitivity, but does not reach the PPV of RRT. This may indicate that while low wall shear stress is necessary for plaque formation, its presence alone is not sufficient to predict future plaque locations. Time dependent factors have to be taken into account as well.


CSSI-PRO: a method for secondary structure type editing, assignment and estimation in proteins using linear combination of backbone chemical shifts.
Estimation of secondary structure in polypeptides is important for studying their structure, folding and dynamics. In NMR spectroscopy, such information is generally obtained after sequence specific resonance assignments are completed. We present here a new methodology for assignment of secondary structure type to spin systems in proteins directly from NMR spectra, without prior knowledge of resonance assignments. The methodology, named Combination of Shifts for Secondary Structure Identification in Proteins (CSSI-PRO), involves detection of specific linear combination of backbone (1)H(alpha) and (13)C' chemical shifts in a two-dimensional (2D) NMR experiment based on G-matrix Fourier transform (GFT) NMR spectroscopy. Such linear combinations of shifts facilitate editing of residues belonging to alpha-helical/beta-strand regions into distinct spectral regions nearly independent of the amino acid type, thereby allowing the estimation of overall secondary structure content of the protein. Comparison of the predicted secondary structure content with those estimated based on their respective 3D structures and/or the method of Chemical Shift Index for 237 proteins gives a correlation of more than 90% and an overall rmsd of 7.0%, which is comparable to other biophysical techniques used for structural characterization of proteins. Taken together, this methodology has a wide range of applications in NMR spectroscopy such as rapid protein structure determination, monitoring conformational changes in protein-folding/ligand-binding studies and automated resonance assignment.


Community health workers adherence to referral guidelines: evidence from studies introducing RDTs in two malaria transmission settings in Uganda.
Many malaria-endemic countries have implemented national community health worker (CHW) programmes to serve remote populations that have poor access to malaria diagnosis and treatment. Despite mounting evidence of CHWs' ability to adhere to malaria rapid diagnostic tests (RDTs) and treatment guidelines, there is limited evidence whether CHWs adhere to the referral guidelines and refer severely ill children for further management. In southwest Uganda, this study examined whether CHWs referred children according to training guidelines and described factors associated with adherence to the referral guideline. A secondary analysis was undertaken of data collected during two cluster-randomized trials conducted between January 2010 and July 2011, one in a moderate-to-high malaria transmission setting and the other in a low malaria transmission setting. All CHWs were trained to prescribe artemisinin-based combination therapy (ACT) and recognize symptoms in children that required immediate referral to the nearest health centre. Intervention arm CHWs had additional training on how to conduct an RDT; CHWs in the control arm used a presumptive diagnosis for malaria using clinical signs and symptoms. CHW treatment registers were reviewed to identify children eligible for referral according to training guidelines (temperature of ≥38.5 °C), to assess whether CHWs adhered to the guidelines and referred them. Factors associated with adherence were examined with logistic regression models. CHWs failed to refer 58.8% of children eligible in the moderate-to-high transmission and 31.2% of children in the low transmission setting. CHWs using RDTs adhered to the referral guidelines more frequently than CHWs not using RDTs (moderate-to-high transmission: 50.1 vs 18.0%, p = 0.003; low transmission: 88.5 vs 44.1%, p < 0.001). In both settings, fewer than 20% of eligible children received pre-referral treatment with rectal artesunate. Children who were prescribed ACT were very unlikely to be referred in both settings (97.7 and 73.3% were not referred in the moderate-to-high and low transmission settings, respectively). In the moderate-to-high transmission setting, day and season of visit were also associated with the likelihood of adherence to the referral guidelines, but not in the low transmission setting. CHW adherence to referral guidelines was poor in both transmission settings. However, training CHWs to use RDT improved correct referral of children with a high fever compared to a presumptive diagnosis using sign and symptoms. As many countries scale up CHW programmes, routine monitoring of reported data should be examined carefully to assess whether CHWs adhere to referral guidelines and take remedial actions where required.


Anodic Voltammetry of Thioacetamide and its Amperometric Determination in Aqueous Media.
TAA is a harmful, presumptive pollutant in tap waters and waste waters. Several alternatives have been tested as new possibilities for the anodic determination of TAA in aqueous solutions, simulated waste waters and tap water. The electrochemical behaviour of thioacetamide (TAA) was investigated at a boron-doped diamond (BDD) electrode both in unbuffered 0.1 M Na₂SO₄ and buffered solutions as supporting electrolytes. The anodic oxidation of TAA showed well-defined limiting currents or current peaks and a good linearity of the amperometric signal vs. concentration plots. The analytical parameters of sensitivity, RSD and LOD, obtained under various experimental conditions, suggest the suitability of the BDD electrode for electroanalytical purposes. Low fouling effects, good reproducibility and stability, as well as the sharpness of the amperometric signals in both unbuffered/ buffered acidic or neutral media, highly superior to those obtained using a glassy carbon (GC) electrode, recommend the unmodified BDD electrode as a promising potential amperometric sensor for environmental applications, regarding the direct anodic determination of TAA in aqueous media.


Therapy of lupus nephritis: lessons learned from clinical research and daily care of patients.
Despite numerous randomized clinical trials over the last three decades for identifying the optimal treatment option for lupus nephritis, renal involvement still significantly impacts the survival and quality of life of patients with lupus and the search for the ideal immunosuppressive regimen is far from complete. The purpose of this review is to summarize the major recent achievements in the field. More specifically, the following topics will be discussed: intravenous cyclophosphamide versus mycophenolate mofetil (MMF) for induction; azathioprine versus MMF for maintenance; targeted therapies. The review will address clues for optimal global care, such as the need for complete initial evaluation, the importance of patient education, the unmasking of non-compliance to therapy, the reason for an early treatment switch in non-responding patients, the need for prolonged immunosuppression, optimal renal protection, and prevention of cardiovascular disease and other comorbidities.


Three-dimensional fiber deposition of cell-laden, viable, patterned constructs for bone tissue printing.
Organ or tissue printing, a novel approach in tissue engineering, creates layered, cell-laden hydrogel scaffolds with a defined three-dimensional (3D) structure and organized cell placement. In applying the concept of tissue printing for the development of vascularized bone grafts, the primary focus lies on combining endothelial progenitors and bone marrow stromal cells (BMSCs). Here we characterize the applicability of 3D fiber deposition with a plotting device, Bioplotter, for the fabrication of spatially organized, cell-laden hydrogel constructs. The viability of printed BMSCs was studied in time, in several hydrogels, and extruded from different needle diameters. Our findings indicate that cells survive the extrusion and that their subsequent viability was not different from that of unprinted cells. The applied extrusion conditions did not affect cell survival, and BMSCs could subsequently differentiate along the osteoblast lineage. Furthermore, we were able to combine two distinct cell populations within a single scaffold by exchanging the printing syringe during deposition, indicating that this 3D fiber deposition system is suited for the development of bone grafts containing multiple cell types.


Delta9-tetrahydrocannabinol inhibits cell cycle progression in human breast cancer cells through Cdc2 regulation.
It has been proposed that cannabinoids are involved in the control of cell fate. Thus, these compounds can modulate proliferation, differentiation, and survival in different manners depending on the cell type and its physiopathologic context. However, little is known about the effect of cannabinoids on the cell cycle, the main process controlling cell fate. Here, we show that Delta(9)-tetrahydrocannabinol (THC), through activation of CB(2) cannabinoid receptors, reduces human breast cancer cell proliferation by blocking the progression of the cell cycle and by inducing apoptosis. In particular, THC arrests cells in G(2)-M via down-regulation of Cdc2, as suggested by the decreased sensitivity to THC acquired by Cdc2-overexpressing cells. Of interest, the proliferation pattern of normal human mammary epithelial cells was much less affected by THC. We also analyzed by real-time quantitative PCR the expression of CB(1) and CB(2) cannabinoid receptors in a series of human breast tumor and nontumor samples. We found a correlation between CB(2) expression and histologic grade of the tumors. There was also an association between CB(2) expression and other markers of prognostic and predictive value, such as estrogen receptor, progesterone receptor, and ERBB2/HER-2 oncogene. Importantly, no significant CB(2) expression was detected in nontumor breast tissue. Taken together, these data might set the bases for a cannabinoid therapy for the management of breast cancer.


Clinical course and factors associated with remission in pemphigus vulgaris patients in Puerto Rico.
To describe and identify those factors associated with remission on pemphigus vulgaris (PV) patients in Puerto Rico. This retrospective cohort study evaluated PV patients followed at the University Puerto Rico (UPR) Bullous Diseases Clinic during the 2000-2010 period. Patients included in the study had clinical and pathologic findings consistent with PV and had a disease duration of at least 3 years. Variables including gender, date of birth, time of disease onset, and date of first partial or complete remission were collected from the medical chart for each study participant. The primary outcome was to determine the number of patients who achieved partial or complete remission. Other secondary outcomes were to identify if partial/complete remission were associated to gender, disease duration, and age at onset of disease. Among 35 patients included in this study, 6 (17%) achieved complete remission and 28 (80%) achieved partial remission. A statistically-significant association was found between duration of disease and remission, predicting a 52% probability of remission after ten years of disease duration. Age at onset of disease showed a trend association with remission, although it was not statistically significant. Gender was not associated with remission. These findings provide insights into the clinical course of PV and can be of value in the management and care of this patient population.


EWSR1-WT1 Target Genes and Therapeutic Options Identified in a Novel DSRCT In Vitro Model.
Desmoplastic small round cell tumor (DSRCT) is a rare and aggressive soft tissue sarcoma with a lack of effective treatment options and a poor prognosis. DSRCT is characterized by a chromosomal translocation, resulting in the EWSR1-WT1 gene fusion. The molecular mechanisms driving DSRCT are poorly understood, and a paucity of preclinical models hampers DSRCT research. Here, we establish a novel primary patient-derived DSRCT in vitro model, recapitulating the original tumor. We find that EWSR1-WT1 expression affects cell shape and cell survival, and we identify downstream target genes of the EWSR1-WT1 fusion. Additionally, this preclinical in vitro model allows for medium-throughput drug screening. We discover sensitivity to several drugs, including compounds targeting RTKs. MERTK, which has been described as a therapeutic target for several malignancies, correlates with EWSR1-WT1 expression. Inhibition of MERTK with the small-molecule inhibitor UNC2025 results in reduced proliferation of DSRCT cells in vitro, suggesting MERTK as a therapeutic target in DSRCT. This study underscores the usefulness of preclinical in vitro models for studying molecular mechanisms and potential therapeutic options.


Ceramic Heads Decrease Metal Release Caused by Head-taper Fretting and Corrosion.
Metal release resulting from taper fretting and corrosion is a clinical concern, because wear and corrosion products may stimulate adverse local tissue reactions. Unimodular hip arthroplasties have a conical taper between the femoral head (head bore taper) and the femoral stem (stem cone taper). The use of ceramic heads has been suggested as a way of reducing the generation of wear and corrosion products from the head bore/stem cone taper junction. A previous semiquantitative study found that ceramic heads had less visual evidence of fretting-corrosion damage compared with CoCr heads; but, to our knowledge, no studies have quantified the volumetric material loss from the head bore and stem cone tapers of a matched cohort of ceramic and metal heads. We asked: (1) Do ceramic heads result in less volume of material loss at the head-stem junction compared with CoCr heads; (2) do stem cone tapers have less volumetric material loss compared with CoCr head bore tapers; (3) do visual fretting-corrosion scores correlate with volumetric material loss; and (4) are device, patient, or intraoperative factors associated with volumetric material loss? A quantitative method was developed to estimate volumetric material loss from the head and stem taper in previously matched cohorts of 50 ceramic and 50 CoCr head-stem pairs retrieved during revision surgery for causes not related to adverse reactions to metal particles. The cohorts were matched according to (1) implantation time, (2) stem flexural rigidity, and (3) lateral offset. Fretting corrosion was assessed visually using a previously published four-point, semiquantitative scoring system. The volumetric loss was measured using a precision roundness machine. Using 24 equally spaced axial traces, the volumetric loss was estimated using a linear least squares fit to interpolate the as-manufactured surfaces. The results of this analysis were considered in the context of device (taper angle clearance, head size, head offset, lateral offset, stem material, and stem surface finish) and patient factors that were obtained from the patients' operative records (implantation time, age at insertion, activity level, and BMI). The cumulative volumetric material losses estimated for the ceramic cohort had a median of 0.0 mm(3) per year (range, 0.0-0.4 mm(3)). The cumulative volumetric material losses estimated for the CoCr cohort had a median of 0.1 mm(3) per year (range, 0.0-8.8 mm(3)). An order of magnitude reduction in volumetric material loss was found when a ceramic head was used instead of a CoCr head (p < 0.0001). In the CoCr cohort, the femoral head bore tapers had a median material loss of 0.02 mm(3) (range, 0.0-8.7 mm(3)) and the stem cone tapers had a median material loss of 0.0 mm(3) (range, 0.0-0.32 mm(3)/year). There was greater material loss from femoral head bore tapers compared with stem cone tapers in the CoCr cohort (p < 0.001). There was a positive correlation between visual scoring and volumetric material loss (Spearman's ρ = 0.67, p < 0.01). Although visual scoring was effective for preliminary screening to separate tapers with no or mild damage from tapers with moderate to severe damage, it was not capable of discriminating in the large range of material loss observed at the taper surfaces with moderate to severe fretting-corrosion damage, indicated with a score of 3 or 4. We observed no correlations between volumetric material loss and device and patient factors. The majority of estimated material loss from the head bore-stem cone junctions resulting from taper fretting and corrosion was from the CoCr head bore tapers as opposed to the stem cone tapers. Additionally, the total material loss from the ceramic cohort showed a reduction in the amount of metal released by an order of magnitude compared with the CoCr cohort. We found that ceramic femoral heads may be an effective means by which to reduce metal release caused by taper fretting and corrosion at the head bore-stem cone modular interface in THAs.


Effect of anthocyanins on expression of matrix metalloproteinase-2 in naproxen-induced gastric ulcers.
Non-steroidal anti-inflammatory drugs cause gastric ulceration through a number of mechanisms including inhibition of PG synthesis, generation of reactive oxygen species (ROS) and induction of apoptosis. Recently, matrix metalloproteinases (MMP) have been suggested to play a crucial role in these mechanisms. The present study investigated the protective effect of anthocyanins isolated from black rice bran (Heugjinjubyeo) against naproxen-induced gastric mucosal injury in rats. The oral administration of anthocyanins (5, 25 or 50 mg/kg body weight) showed significant protection against naproxen (80 mg/kg body weight)-induced gastric ulcer and inhibited lipid peroxidation in the gastric mucosa. In addition, pretreatment with anthocyanins resulted in a significant increase in the activities of radical-scavenging enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Also biochemical and zymographic analyses suggested that the administration of anthocyanins gives a significant protection against naproxen-induced gastric antral ulcer through scavenging ROS and regulation of matrix metalloproteinase-2 (MMP-2) activity. The results of intracellular radical activation show that anthocyanins suppress the generation of intracellular ROS and attenuate the suppression of MMP-2 activity by naproxen. These results suggest that anthocyanins extracted from black rice may offer potential remedy of gastric antral ulceration.


A theoretical study of low-lying singlet and triplet excited states of quinazoline, quinoxaline and phthalazine: insight into triplet formation.
Quinazoline, quinoxaline and phthalazine are nitrogen containing heterocyclic aromatic molecules which belong to the class diazanaphthalenes. These isomers have low-lying nπ* and naphthalene-like ππ* states that interact via spin-orbit coupling. In this contribution, we study their structure and electronic states by means of a coupled-cluster method. The computed properties are compared to those of cinnoline which were obtained in our previous study [Etinski et al., Phys. Chem. Chem. Phys., 2014, 16, 4740]. The excited state features of these isomers are dependent on the position of the nitrogen atoms. We find that quinazoline and quinoxaline exhibit similarities in the ordering and character of the excited states. In contrast, a marked difference in the electronic and geometric structures of the lowest excited triplet states of cinnoline and phthalazine is noticed, although both are orthodiazanaphthalenes. Our findings suggest that the S1 [radiolysis arrow - arrow with voltage kink] T1 channel is responsible for the rapid intersystem crossing in quinazoline and quinoxaline, whereas the S1 [radiolysis arrow - arrow with voltage kink] T2 pathway is active in phthalazine.


A comprehensive system for evaluation of remote sequence similarity detection.
Accurate and sensitive performance evaluation is crucial for both effective development of better structure prediction methods based on sequence similarity, and for the comparative analysis of existing methods. Up to date, there has been no satisfactory comprehensive evaluation method that (i) is based on a large and statistically unbiased set of proteins with clearly defined relationships; and (ii) covers all performance aspects of sequence-based structure predictors, such as sensitivity and specificity, alignment accuracy and coverage, and structure template quality. With the aim of designing such a method, we (i) select a statistically balanced set of divergent protein domains from SCOP, and define similarity relationships for the majority of these domains by complementing the best of information available in SCOP with a rigorous SVM-based algorithm; and (ii) develop protocols for the assessment of similarity detection and alignment quality from several complementary perspectives. The evaluation of similarity detection is based on ROC-like curves and includes several complementary approaches to the definition of true/false positives. Reference-dependent approaches use the 'gold standard' of pre-defined domain relationships and structure-based alignments. Reference-independent approaches assess the quality of structural match predicted by the sequence alignment, with respect to the whole domain length (global mode) or to the aligned region only (local mode). Similarly, the evaluation of alignment quality includes several reference-dependent and -independent measures, in global and local modes. As an illustration, we use our benchmark to compare the performance of several methods for the detection of remote sequence similarities, and show that different aspects of evaluation reveal different properties of the evaluated methods, highlighting their advantages, weaknesses, and potential for further development. The presented benchmark provides a new tool for a statistically unbiased assessment of methods for remote sequence similarity detection, from various complementary perspectives. This tool should be useful both for users choosing the best method for a given purpose, and for developers designing new, more powerful methods. The benchmark set, reference alignments, and evaluation codes can be downloaded from ftp://iole.swmed.edu/pub/evaluation/.


Reduced substrate oxidation in postischemic myocardium: 13C and 31P NMR analyses.
13C and 31P nuclear magnetic resonance (NMR) spectra were used to assess substrate oxidation and high-energy phosphates in postischemic (PI) isolated rabbit hearts. Phosphocreatine (PCr) increased in nonischemic controls on switching from glucose perfusion to either 2.5 mM [3-13C]pyruvate (120%, n = 7) or [2-13C]acetate (114%, n = 8, P less than 0.05). ATP content, oxygen consumption (MVO2), and hemodynamics (dP/dt) were not affected by substrate availability in control or PI hearts. dP/dt was 40-60% lower in PI hearts during reperfusion after 10 min ischemia. Hearts reperfused with either pyruvate (n = 11) or acetate (n = 8) regained preischemic PCr levels within 45 s. Steady-state ATP levels were 55-70% of preischemia with pyruvate and 52-60% with acetate. Percent maximum [4-13C]glutamate signal showed reduced conversion of pyruvate to glutamate via the tricarboxylic acid (TCA) cycle at 4-min reperfusion (PI = 24 +/- 4%, means +/- SE; Control = 48 +/- 4%). The increase in 13C signal from the C-4 position of glutamate was similar to control hearts within 10.5 min. The increase in [4-13C]glutamate signal from acetate was not different between PI and control hearts. The ratio of [2-13C]Glu:[4-13C]Glu, reflecting TCA cycle activity, was reduced in PI hearts with acetate for at least 10 min (Control = 0.76 +/- 0.03; PI = 0.51 +/- 0.09) until steady state was reached. Despite rapid recovery of oxidative phosphorylation, contractility remained impaired and substrate oxidation was significantly slowed in postischemic hearts.


Evaluation of a multidisciplinary pediatric mock trauma code educational initiative: a pilot study.
Our aim was to develop and evaluate a multidisciplinary pediatric mock trauma code orientation program for residents on their pediatric hospital rotation. A before and after evaluation of trauma team residents from various programs was conducted. The 1-hour educational session consisted of a 15-minute mock trauma code, a debriefing and teaching intervention, and then a new, postintervention mock trauma code. Before and after each session, residents completed a self-assessment questionnaire. All codes were videotaped and later evaluated by two blinded observers using a standardized evaluation tool to assess patient management and team functioning. Thirty-seven prequestionnaire (pre) and postquestionnaire (post) pairs were completed. Residents reported a significant improvement in their (1) comfort in managing pediatric trauma (median pre 3, post 5, p < 0.001); (2) understanding of their role on the trauma team (median pre 4.5, post 6, p < 0.001); (3) familiarity with the resuscitation room (median pre 4, post 5, p = 0.001); (4) comfort with procedural skills (median pre 4, post 5, p = 0.001); (5) awareness of pediatric trauma resuscitation management priorities (median pre 5, post 6, p = 0.007). Postintervention, residents reported lower knowledge scores in locating equipment in the resuscitation room (p < 0.001). There was no significant difference in team performance on the videotaped assessments (premean score = 79.8, postmean score = 79.5). The pediatric mock trauma code educational initiative improved residents' self-reported confidence, knowledge, and comfort level in managing pediatric trauma. The experience also raised residents' awareness of knowledge gaps. We were unable to measure a significant change in team functioning post intervention.


Immobilization of free-ranging nine-banded and great long-nosed armadillos with three anesthetic combinations.
Nine-banded (n = 47) and great (n = 31) long-nosed armadillos (Dasypus novemcinctus and Dasypus kappleri) were immobilized for clinical examination and collection of biological samples as part of a wildlife rescue during the filling of a hydroelectric dam (Petit Saut, French Guiana) from May 1994 to April 1995. Three intramuscular (i.m.) anesthetic combinations were evaluated: (1) tiletamine/zolazepam (T/Z) at a dose of 8.5 mg/kg in 12 nine-banded long-nosed armadillos (NBA) and 10 great long-nosed armadillos (GLA), (2) ketamine at 40 mg/kg combined with xylazine at 1.0 mg/kg (K/X) in 18 NBA and nine GLA, and (3) ketamine at 7.5 mg/kg combined with medetomidine at 75 microng/kg (K/M) in 17 NBA and 12 GLA, antagonized by 375 microg/kg atipamezole. Induction was smooth, ranged from mean +/- SD = 2.8+/-0.6 to 4.3+/-1.8 min, and did not differ significantly between protocols, species, or sex. In NBA, immobilization time ranged from 43.8+/-27.8 to 66.5+/-40.0 min and did not differ between protocols or sex. Muscle relaxation was judged to be better with K/X and K/M versus T/Z. In GLA, the response to the anesthetic protocols was more variable and immobilization time ranged from 30.4+/-6.2 to 98.4+/-33.7 min. The main difference was observed in GLA females receiving the T/Z combination, in which immobilization time was significantly longer versus males, but also versus GLA K/M group, and versus NBA T/Z group. Effects on body temperature, heart rate and respiratory rate were limited. Thirty six to 50% of the individuals showed hypoxemia (SpO2 < 85%) throughout anesthesia and values <80% also were recorded but the hypoxemia was not associated with clinical signs. With T/Z and K/X, recovery was irregular and prolonged up to 2 to 3 hr in some individuals. In K/M groups, first standing was observed 1.0 to 16.4 min after i.m. atipamezole injection without adverse effects. Finally, the three anesthetic combinations used in this study were effective and safe agents for 30 to 40 min immobilizations including minor surgery procedures. The ability to antagonize the medetomidine-induced sedation with atipamezole significantly reduces the recovery time, making the K/M combination preferable, especially in field conditions.


Interleukin 4 or oncostatin M induces a prolonged increase in P-selectin mRNA and protein in human endothelial cells.
During acute inflammation, P-selectin is transiently mobilized from Weibel-Palade bodies to the surface of histamine-activated endothelial cells, where it mediates rolling adhesion of neutrophils under hydrodynamic flow. During chronic or allergic inflammation, sustained expression of P-selectin on the endothelial cell surface has been observed. We found that the cytokines interleukin 4 (IL-4) or oncostatin M (OSM) induced a five- to ninefold increase in P-selectin messenger RNA (mRNA) in human umbilical vein endothelial cells (HUVEC) that persisted as long as 72 h. IL-4 elevated P-selectin mRNA by increasing its transcription rate rather than by prolonging its already long half-life. Stimulation of P-selectin transcription by IL-4 or OSM required new protein synthesis and tyrosine phosphorylation of cellular proteins. Tumor necrosis factor alpha, IL-1 beta, lipopolysaccharide, or IL-3 did not increase P-selectin mRNA in HUVEC, and did not augment the IL-4-induced increase in P-selectin transcripts. IL-4 or OSM increased P-selectin protein on the cell surface as well as in Weibel-Palade bodies. Under flow conditions, neutrophils rolled on P-selectin expressed by IL-4-treated HUVEC, and even more neutrophils rolled on P-selectin after IL-4-treated HUVEC were stimulated with histamine. These data demonstrate that IL-4 or OSM stimulates endothelial cells to synthesize more P-selectin over prolonged periods. The increased expression of P-selectin may facilitate the emigration of leukocytes into sites of chronic or allergic inflammation.


Electrically detected magnetic resonance in a W-band microwave cavity.
We describe a low-temperature sample probe for the electrical detection of magnetic resonance in a resonant W-band (94 GHz) microwave cavity. The advantages of this approach are demonstrated by experiments on silicon field-effect transistors. A comparison with conventional low-frequency measurements at X-band (9.7 GHz) on the same devices reveals an up to 100-fold enhancement of the signal intensity. In addition, resonance lines that are unresolved at X-band are clearly separated in the W-band measurements. Electrically detected magnetic resonance at high magnetic fields and high microwave frequencies is therefore a very sensitive technique for studying electron spins with an enhanced spectral resolution and sensitivity.


Expression of functional bacterial undecaprenyl pyrophosphate synthase in the yeast rer2{Delta} mutant and CHO cells.
During evolution the average chain length of polyisoprenoid glycosyl carrier lipids increased from C55 (prokaryotes) to C75 (yeast) to C95 (mammalian cells). In this study, the ability of the E. coli enzyme, undecaprenyl pyrophosphate synthase (UPPS), to complement the loss of the yeast cis-isoprenyltransferase in the rer2Δ mutant was tested to determine if (55)dolichyl phosphate (Dol-P) could functionally substitute in the protein N-glycosylation pathway for (75)Dol-P, the normal isoprenologue synthesized in S. cerevisiae. First, expression of UPPS in the yeast mutant was found to complement the growth and the hypoglycosylation of carboxypeptidase Y defects suggesting that the (55)polyprenyl-P-P intermediate was converted to (55)Dol-P and that (55)Dol-P could effectively substitute for (75)Dol-P in the biosynthesis and function of Man-P-Dol, Glc-P-Dol and Glc(3)Man(9)GlcNAc(2)-P-P-Dol (mature DLO) in the protein N-glycosylation pathway and glycosylphosphatidylinositol anchor assembly. In support of this conclusion, mutant cells expressing UPPS (1) synthesized (55)Dol-P based on MS analysis, (2) utilized (55)Dol-P to form Man-P-(55)Dol in vitro and in vivo, and (3) synthesized N-linked glycoproteins at virtually normal rates as assessed by metabolic labeling with [(3)H]mannose. In addition, an N-terminal GFP-tagged construct of UPPS was shown to localize to the endoplasmic reticulum of Chinese hamster ovary cells. Consistent with the synthesis of (55)Dol-P by the transfected cells, microsomes from the transfected cells synthesized the [(14)C](55)polyprenyl-P-P intermediate when incubated with [(14)C]isopentenyl pyrophosphate and [(3)H]Man-P-(55)Dol when incubated with GDP-[(3)H]Man. These results indicate that (C55)polyisoprenoid chains, significantly shorter than the natural glycosyl carrier lipid, can function in the transbilayer movement of DLOs in the endoplasmic reticulum of yeast and mammalian cells, and that conserved sequences in the cis-isoprenyltransferases are recognized by, yet to be identified, binding partners in the endoplasmic reticulum of mammalian cells.


Value of throat swab in diagnosis of melioidosis.
Throat swab (TS) cultures were performed for 1,011 patients with melioidosis and 3,524 healthy subjects or patients with other diseases. The specificity of TS culture for the diagnosis of melioidosis was 100%, and the overall sensitivity was 36% (24% for sputum-negative patients and 79% for sputum-positive patients). Direct plating of the TS specimen on Ashdown's medium was rapid (colonies were usually evident within 24 h) but only 63% sensitive compared to the results of primary culture in a selective broth. A throat swab should be cultured in all cases of suspected melioidosis.


Novel insight into the genetic basis of high-altitude pulmonary hypertension in Kyrgyz highlanders.
The Central Asian Kyrgyz highland population provides a unique opportunity to address genetic diversity and understand the genetic mechanisms underlying high-altitude pulmonary hypertension (HAPH). Although a significant fraction of the population is unaffected, there are susceptible individuals who display HAPH in the absence of any lung, cardiac or hematologic disease. We report herein the analysis of the whole-genome sequencing of healthy individuals compared with HAPH patients and other controls (total n = 33). Genome scans reveal selection signals in various regions, encompassing multiple genes from the first whole-genome sequences focusing on HAPH. We show here evidence of three candidate genes MTMR4, TMOD3 and VCAM1 that are functionally associated with well-known molecular and pathophysiological processes and which likely lead to HAPH in this population. These processes are (a) dysfunctional BMP signaling, (b) disrupted tissue repair processes and (c) abnormal endothelial cell function. Whole-genome sequence of well-characterized patients and controls and using multiple statistical tools uncovered novel candidate genes that belong to pathways central to the pathogenesis of HAPH. These studies on high-altitude human populations are pertinent to the understanding of sea level diseases involving hypoxia as a main element of their pathophysiology.


Intraspecific vertical stratification as a mate-finding mechanism in tropical cockroaches.
Cockroaches in a tropical forest stratify vertically both inter- and intraspecifically along micrometeorological gradients. At night, low wind speeds and unstable atmospheric conditions result in efficient vertical mixing of the air near the ground. Convective ascent of warm air imparts directionality to the pheromone- dispersion process. The occurrence of males at greater heights than pheromone- emitting conspecific females appears to be a mate-finding strategy.