Validating the EMCV IRES Secondary Structure with Structure-Function Analysis.
The encephalomyocarditis virus internal ribosome entry site (EMCV IRES) is a structured RNA sequence found in the 5' UTR of the genomic RNA of the encephalomyocarditis virus. The EMCV IRES structure facilitates efficient translation initiation without needing a 5' m7G cap or the cap-binding protein eIF4E. The secondary structure of IRES has been the subject of several previous studies, and a number of different structural models have been proposed. Though some domains of the IRES are conserved across the different secondary structure models, domain I of the IRES varies greatly across them. A literature comparison led to the identification of three regions of interest that display structural heterogeneity within past secondary structure models. To test the accuracy of the secondary structure models in these regions, we employed mutational analysis and SHAPE probing. Mutational analysis revealed that two helical regions within the identified regions of interest are important for IRES translation. These helical regions are consistent with only one of the structure predictions in the literature and do not form in EMCV IRES structures predicted using modern secondary structure prediction methods. The importance of these regions is further supported by multiple SHAPE protections when probing was performed after in vitro translation, indicating that these regions are involved in the IRES translation complex. This work validates a published structure and demonstrates the importance of domain I during EMCV IRES translation initiation.


Design and synthesis of a library of C2-substituted sulfamidoadenosines to probe bacterial permeability.
Antibiotic resistance is a major threat to public health, and Gram-negative bacteria pose a particular challenge due to their combination of a low permeability cell envelope and efflux pumps. Our limited understanding of the chemical rules for overcoming these barriers represents a major obstacle in antibacterial drug discovery. Several recent efforts to address this problem have involved screening compound libraries for accumulation in bacteria in order to understand the structural properties required for Gram-negative permeability. Toward this end, we used cheminformatic analysis to design a library of sulfamidoadenosines (AMSN) having diverse substituents at the adenine C2 position. An efficient synthetic route was developed with installation of a uniform cross-coupling reagent set using Sonogashira and Suzuki reactions of a C2-iodide. The potential utility of these compounds was demonstrated by pilot analysis of selected analogues for accumulation in Escherichia coli.


Pharmacotherapy in anorexia nervosa: A Danish nation-wide register-based study.
No medications have been indicated for the treatment of anorexia nervosa (AN). Nonetheless, individuals with AN are frequently treated pharmacologically. The present study maps nationwide pharmacotherapy two years before to five years after first AN diagnosis. We identified all medication prescriptions in a national register-based study of patients with a first diagnosis of AN between 1998 and 2011, and age and gender matched controls (1:10). Medication classes were compared using odds ratios (OR) between patients and controls; between patients below and above 15 years; between patients with and without comorbidity; and between those diagnosed before or after 2005. The odds of pharmacotherapy were increased in patients for all classes of medication except a small residual class. Highest odds were found for alimentary (OR 2.8, p < 0.001) and psychopharmacological (OR 5.5, p < 0.001) medication. The former peaked one year prior to first diagnosis and the latter one year after. Older patients had increased risk of almost all medication classes with cardiovascular medication showing a fivefold OR (p < 0.001). Patients with psychiatric comorbidity had a threefold OR for psychopharmacological medication (p < 0.001) compared to patients without psychiatric comorbidity. Calendar year showed few and small differences. The extended use of all medication classes both prior to and after first diagnosis of AN highlights the severe cause and complexity of AN. The results encourage clinical caution of pharmacotherapy, highlight the need for pharmacotherapy guidelines for AN, and emphasize the urgency of research in pharmacotherapy in AN.


[Construct validity of a Portuguese version of the Female Sexual Function Index].
This study aimed to evaluate the construct validity of a version of the Female Sexual Function Index (FSFI). The version was inserted into a multidimensional questionnaire and applied face-to-face to 235 sterilized women. Non-response rate was 1.7%. The sample proved to be highly homogeneous, with low income and education. Cronbach's alpha of 0.948 indicated high internal consistency. Factor analysis showed that the instrument was measuring four factors: desire/arousal, lubrication, orgasm/satisfaction, and pain. This may reflect characteristics of the instrument itself, female sexual response, or even the sample's cultural peculiarities, which can affect understanding of the questions or concepts with which the instrument deals. This study emphasizes the need for further research in the general population to determine the psychometric properties of the FSFI, such as its factor composition and definition of appropriate cut-off points for the Brazilian population.


Stacking the deck for success in interprofessional collaboration.
The practice of interprofessional collaboration is increasingly being recognized as an important skill for educators, health professionals, and social service providers. Collaboration involves team building and developing integrated service delivery mechanisms to improve outcomes for recipients of health, education, and social services. The focus on health promotion and illness prevention has led to an increasing demand for health educators to become involved in these collaboratives as members and more often as facilitators. Key concepts associated with early stages of collaborative development are described, with a focus on forming a group identity and weathering the conflicts associated with task and personnel issues. Descriptions and instructions for facilitating activities to move the group successfully through these initial stages are described.


A Systematic Literature Review of Patients With Carotid Web and Acute Ischemic Stroke.
Background and Purpose- Carotid web (CW) is a rare form of focal fibromuscular dysplasia defined as an abnormal shelf-like projection of intimal fibrous tissue into the carotid bulb. It is theorized that CW leads to ischemic stroke secondary to blood flow stasis and subsequent embolization. The natural history and optimal management of CW are unclear. To address this knowledge gap, we performed a systematic literature review (SLR) of CW. Methods- Our librarians performed a SLR for CW and related terminology. Patient-level demographics, stroke risk factors, neuroimaging findings, stroke recurrence or stroke free-duration, and treatment modality were extracted. We used descriptive statistics to characterize our results. When specific patient-level metrics were not reported, the denominators for reporting percentage calculations were adjusted accordingly. Results- Our literature search produced 1150 articles. Thirty-seven articles including 158 patients (median age 46 years [range 16-85], 68% women, 76% symptomatic) met entry criteria and were included in our SLR. Of the symptomatic CW patients: 57% did not have stroke risk factors, 56% who received medical therapy had recurrent stroke (median 12 months, range 0-97), and 72% were ultimately treated with carotid revascularization (50% carotid stenting, 50% carotid endarterectomy). There were no periprocedural complications or recurrent strokes in carotid revascularization patients. Conclusions- CW leads to ischemic stroke in younger patients without conventional stroke risk factors. We found a high stroke recurrence rate in medically managed symptomatic CW patients, whereas carotid revascularization effectively prevented recurrent stroke. Our findings should be interpreted with caution because of risk of publication and reporting bias.


Fetal and neonatal exposure to nicotine disrupts postnatal lung development in rats: role of VEGF and its receptors.
Many women are unable to quit smoking during pregnancy and therefore are prescribed drugs, including nicotine (nicotine replacement therapy [NRT]), to aid with smoking cessation. However, the consequences to the offspring of pregnant NRT users have not been well studied. The goals of this study were to determine the consequences of fetal and neonatal exposure to nicotine on lung development and function. Female rats were exposed to nicotine for 2 weeks prior to mating until weaning. Lungs were collected from saline and nicotine-treated rats from birth to adulthood to assess postnatal lung structure and function. Although nicotine exposure altered alveolarization at weaning, an effect that resolved by adulthood, it did not affect lung function at any of the ages investigated. However, nicotine exposure significantly decreased lung vascularization. The current study suggests that perinatal exposure to nicotine alters lung development, an effect which may be mediated via decreased vascular endothelial growth factor (VEGF) signaling.


Sensitivity of Colorectal Cancer to Arginine Deprivation Therapy is Shaped by Differential Expression of Urea Cycle Enzymes.
Tumors deficient in the urea cycle enzymes argininosuccinate synthase-1 (ASS1) and ornithine transcarbamylase (OTC) are unable to synthesize arginine and can be targeted using arginine-deprivation therapy. Here, we show that colorectal cancers (CRCs) display negligible expression of OTC and, in subset of cases, ASS1 proteins. CRC cells fail to grow in arginine-free medium and dietary arginine deprivation slows growth of cancer cells implanted into immunocompromised mice. Moreover, we report that clinically-formulated arginine-degrading enzymes are effective anticancer drugs in CRC. Pegylated arginine deiminase (ADI-PEG20), which degrades arginine to citrulline and ammonia, affects growth of ASS1-negative cells, whereas recombinant human arginase-1 (rhArg1peg5000), which degrades arginine into urea and ornithine, is effective against a broad spectrum of OTC-negative CRC cell lines. This reflects the inability of CRC cells to recycle citrulline and ornithine into the urea cycle. Finally, we show that arginase antagonizes chemotherapeutic drugs oxaliplatin and 5-fluorouracil (5-FU), whereas ADI-PEG20 synergizes with oxaliplatin in ASS1-negative cell lines and appears to interact with 5-fluorouracil independently of ASS1 status. Overall, we conclude that CRC is amenable to arginine-deprivation therapy, but we warrant caution when combining arginine deprivation with standard chemotherapy.


Dendritic Cell-Based Immunotherapy of Acute Myeloid Leukemia.
Acute myeloid leukemia (AML) is a type of blood cancer characterized by the uncontrolled clonal proliferation of myeloid hematopoietic progenitor cells in the bone marrow. The outcome of AML is poor, with five-year overall survival rates of less than 10% for the predominant group of patients older than 65 years. One of the main reasons for this poor outcome is that the majority of AML patients will relapse, even after they have attained complete remission by chemotherapy. Chemotherapy, supplemented with allogeneic hematopoietic stem cell transplantation in patients at high risk of relapse, is still the cornerstone of current AML treatment. Both therapies are, however, associated with significant morbidity and mortality. These observations illustrate the need for more effective and less toxic treatment options, especially in elderly AML and have fostered the development of novel immune-based strategies to treat AML. One of these strategies involves the use of a special type of immune cells, the dendritic cells (DCs). As central orchestrators of the immune system, DCs are key to the induction of anti-leukemia immunity. In this review, we provide an update of the clinical experience that has been obtained so far with this form of immunotherapy in patients with AML.


Bivalirudin.
Bivalirudin is a direct thrombin inhibitor (DTI) frequently used for anticoagulation in the setting of invasive cardiology, particularly percutaneous coronary intervention (PCI). Bivalirudin has a unique pharmacologic profile: unlike other marketed DTIs, it undergoes predominant non-organ elimination (proteolysis), and has the shortest half-life (approximately 25 min). Its affinity for thrombin is intermediate between that of lepirudin (highest) and argatroban (lowest)--this helps explain why it interferes with functional clotting assays to an extent intermediate between that achieved by these two other DTIs. This effect is best known for the PT (INR)--higher affinity for thrombin corresponds to lower molar DTI requirements to prolong the APTT; in turn, lower concentrations required for APTT prolongation (and, presumably, in-vivo effect) result in reduced PT (INR) prolongation. Bivalirudin is primarily used for its first FDA-approved indication, namely anticoagulation during percutaneous transluminal coronary angioplasty ("balloon angioplasty"), the most frequent type of PCI. Bivalirudin is also indicated for PCI with provisional use of glycoprotein IIb/IIIa antagonist therapy, and for patients with, or at risk of, heparin-induced thrombocytopenia (HIT), or HIT with thrombosis syndrome (HITTS), undergoing PCI. The bivalirudin development program has used a "quadruple" endpoint comprising a "triple" efficacy endpoint plus major bleeding - this approach anticipated the subsequent emphasis on strategies to improve clinical outcomes through bleeding reduction. Besides summarizing the key trials evaluating bivalirudin use for acute coronary syndrome (especially employing PCI), we review also the studies of bivalirudin as anticoagulant for "on-" and "off-pump" cardiac surgery, including both HIT and non-HIT situations.


Implementation of Provider-Focused Education to Improve Laboratory Screening for Pediatric Patients With Elevated Body Mass Index in Primary Care: A Quality Improvement Project.
Children with elevated body mass index (BMI) do not consistently receive recommended laboratory screenings. This project aimed to increase provider screening rates and knowledge of screening guidelines for this population. This project utilized the Rosswurm and Larrabee evidence-based practice model. Providers completed education with pretest and posttest design. Laboratory screening rates were measured with retrospective chart reviews, and a project satisfaction survey was conducted. Nine (82%) of 11 providers showed an increase in knowledge of screening for children with elevated BMI. Laboratory screening increased (27% to 39%) above the preintervention median (25%) for children with obesity and was above the median (22%) for one month of the project (15% to 26%) for children with overweight. Project results suggest education improved knowledge and compliance with guidelines for laboratory screening of children with an elevated BMI. Asynchronous education and sharing of compliance rates are adaptable to similar quality improvement projects.


Thermal diffusion and molecular diffusion values for some alkane mixtures: a comparison between thermogravitational column and thermal diffusion forced rayleigh scattering.
In the present work we studied the thermal diffusion behavior of n-decane in various alkanes by thermogravitational column (TC) technique and the thermal diffusion forced Rayleigh scattering (TDFRS) method. The investigated lighter alkanes compared to n-decane are n-pentane, n-hexane, n-heptane, n-octane, and the heavier ones are n-tetradecane, n-pentadecane, n-hexadecane, n-heptadecane, n-octadecane, and n-eicosane. The binary mixture n-decane/ n-pentane we investigated at several different concentrations; all other mixtures were only investigated at a mass fraction of 50%. Even for the volatile n-pentane/ n-decane mixture the deviations between the thermal diffusion coefficients determined by the different methods agreed within the error bars. Typically the agreement between the two methods was in the order of 5%. In comparison to recently published TC and TDFRS data we found deviations in the order of 30% up to 40%. We analyze and discuss the possible reasons for the discrepancies for the present and the past publications.


The combination of exercise training and alpha-lipoic acid treatment has therapeutic effects on the pathogenic phenotypes of Alzheimer's disease in NSE/APPsw-transgenic mice.
Exercise training was suggested as a practical therapeutic strategy for human subjects suffering from Alzheimer's disease (AD) in our previous study. Therefore, the purpose of this study was to investigate the effects of combining exercise training with the administration of antioxidants on the pathological phenotype of AD. To accomplish this, non-transgenic mice (Non-Tg) and NSE/APPsw Tg mice were treated with alpha-lipoic acid and treadmill exercised for 16 weeks, after which their brains were evaluated to determine whether any changes in the pathological phenotype-related factors occurred. The results indicated that (i) the combination-applied (COMA) Tg group with exercise training (ET) and alpha-lipoic acid administration (LA) showed ameliorated spatial learning and memory compared to the sedentary (SED)-Tg and single-treatment groups; (ii) there were no differences in the level of Abeta-42 peptides across groups; (iii) the level of glucose transporter-1 and brain-derived neurotrophic factor proteins were highly increased in the COMA group, (iv) ET and LA did not induce a synergistic effect on the expression of heat shock protein-70 and apoptotic proteins including Bax and caspase-3; (v) the levels of SOD-1 and CAT suppressing oxidative stress were extensively higher in the COMA than in the single-treated groups and (vi) there were no significant differences across groups regarding these serum characteristics, although these levels were lower than the SED-Tg group. Taken together, these results suggest that the combination with ET and LA may contribute to protect the neuron injury induced by Abeta peptides and may be considered an effective therapeutic strategy for human subjects suffering from AD.


Consistent pollen nutritional intake drives bumble bee (Bombus impatiens) colony growth and reproduction across different habitats.
Foraging behavior is a critical adaptation by insects to obtain appropriate nutrients from the environment for development and fitness. Bumble bees (Bombus spp.) form annual colonies which must rapidly increase their worker populations to support rearing reproductive individuals before the end of the season. Therefore, colony growth and reproduction should be dependent on the quality and quantity of pollen resources in the surrounding landscape. Our previous research found that B. impatiens foraging preferences to different plant species were shaped by pollen protein:lipid nutritional ratios (P:L), with foragers preferring pollen species with a ~5:1 P:L ratio. In this study, we placed B. impatiens colonies in three different habitats (forest, forest edge, and valley) to determine whether pollen nutritional quality collected by the colonies differed between areas that may differ in resource abundance and diversity. We found that habitat did not influence the collected pollen nutritional quality, with colonies in all three habitats collecting pollen averaging a 4:1 P:L ratio. Furthermore, there was no difference in the nutritional quality of the pollen collected by colonies that successfully reared reproductives and those that did not. We found however, that "nutritional intake," calculated as the colony-level intake rate of nutrient quantities (protein, lipid, and sugar), was strongly related to colony growth and reproductive output. Therefore, we conclude that B. impatiens colony performance is a function of the abundance of nutritionally appropriate floral resources in the surrounding landscape. Because we did not comprehensively evaluate the nutrition provided by the plant communities in each habitat, it remains to be determined how B. impatiens polylectic foraging strategies helps them select among the available pollen nutritional landscape in a variety of plant communities to obtain a balance of key macronutrients.


The art of noticing: essential to nursing practice.
Noticing is integral to the everyday practice of nurses; it is the pre-cursor for clinical reasoning, informing judgement and the basis of care. By noticing the nurse can pre-empt possible risks or support subtle changes towards recovery. Noticing can be the activity that stimulates action before words are exchanged, pre-empting need. In this article, the art of noticing is explored in relation to nursing practice and how the failure to notice can have serious consequences for those in care.


Temporal and spectral structure of the infrared pulse during the high order harmonic generation.
We present, for the first time, the complete pulse characterization of the infrared pulse after generating harmonics. A systematic study of the high harmonic generation process, and the generating infrared pulse characterization, has been done by changing the focus-gas-jet relative position. We have concluded, supported by nonlinear propagation simulations, that there is a correlation between the spectral and temporal nonlinear evolution of the infrared generating field and the structures shown in the harmonic signal. We have identified two different pressure regimes: the low pressure regime, characterized by the effects produced by the plasma generated by the infrared pulse, and the high pressure regime where the plasma and the Kerr effect generated by the infrared field are both present. These observations highlight the important role played by the nonlinear propagation of the generating field in the high harmonic generation context.


Surgical and nonsurgical correction of uterine torsion in New World camelids: 20 cases (1990-1996).
To report clinical findings for New World comelids with uterine torsion and to compare results of 3 methods of correction. Retrospective case series. 11 llamas and 3 alpacas with 20 uterine torsions. Information concerning history, clinical signs, management, and postpartum complications was retrieved from medical records. Information concerning subsequent reproductive performance was obtained by telephone interview of owners. Uterine torsion was corrected by celiotomy (n = 7); transvaginal manipulation (5), or rolling the dam (8). Direction of 19 of 20 torsions was clockwise when viewed from the rear. Retention of fetal membranes was reported for 5 camelids that underwent celiotomy, but was not reported in camelids after nonsurgical correction. The uterus prolapsed in 1 llama that underwent celiotomy and in another that underwent the rolling technique. Although 2 camelids that underwent celiotomy subsequently failed to conceive, all camelids treated by nonsurgical techniques conceived. Uterine torsion in camelids may be diagnosed by methods similar to those used in cattle. Surgical and nonsurgical methods can be used to correct torsion, and postpartum complications are rare when torsion is corrected by a nonsurgical method.


Comet 41P/Tuttle-Giacobini-Kresak, 45P/Honda-Mrkos-Pajdusakova, and 46P/Wirtanen: Water Production Activity over 21 Years with SOHO/SWAN.
In 2017, 2018, and 2019, comets 46P/Wirtanen, 45P/Honda-Mrkos-Pajdusakova, and 41P/Tuttle-Giacobini-Kresak all had perihelion passages. Their hydrogen comae were observed by the Solar Wind ANisotropies (SWAN) all-sky hydrogen Lyman-alpha camera on the SOlar and Heliospheric Observer (SOHO) satellite: comet 46P for the fourth time and comets 45P and 41P for the third time each since 1997. Comet 46P/Wirtanen is one of a small class of so-called hyperactive comets whose gas production rates belie their small size. This comet was the original target comet of the Rosetta mission. The Solar Wind ANisotropies (SWAN) all-sky hydrogen Lyman-alpha camera on the SOlar and Heliospheric Observer (SOHO) satellite observed the hydrogen coma of comet 46P/Wirtanen during the apparitions of 1997, 2002, 2008, and 2018. Over the 22 years, the activity decreased and its variation with heliocentric distance has changed markedly in a way very similar to that of another hyperactive comet, 103P/Hartley 2. Comet 45P/Honda-Mrkos-Pajdusakova was observed by SWAN during its perihelion apparitions of 2001, 2011, and 2017. Over this time period the activity level has remained remarkably similar, with no long-term fading or abrupt decreases. Comet 41P/Tuttle-Giacobini-Kresak was observed by SWAN in its perihelion apparitions of 2001, 2006, and 2017 and has decreased in activity markedly over the same time period. In 1973 it was known for large outbursts, which continued during the 2001 (2 outbursts) and 2006 (1 outburst) apparitions. However, over the 2001 to 2017 time period covered by the SOHO/SWAN observations the water production rates have greatly decreased by factors of 10-30 over corresponding times during its orbit.


Medullary neural circuit regeneration after trigeminal nerve transection.
The inferior alveolar nerve (IAN) comprises several types of sensory fibers. To clarify whether each type of primary afferent is regenerated comparably after injury, we developed a model of complete IAN transection (IANX) in mice. A retrograde tracer, fluoro-gold, injected into the mental skin was transferred to the cell bodies of a subset of isolectin B4 (IB4)-binding (non-peptidergic C) or CGRP-positive (peptidergic C) neurons at 2 weeks post-axotomy, indicating that the injured C afferents had regenerated anatomically. IANX led to a decrease of IB4-binding and CGRP immunoreactivity (IR) in the trigeminal ganglion (TG) and within the trigeminal spinal subnucleus caudalis (Vc) (i.e. terminals of the central branch of TG neurons). Two weeks after IANX, the reduction in IB4-binding activity and CGRP expression in the TG recovered to the control level; however, IB4-binding within the Vc did not, suggesting that central branch non-peptidergic neurons remained impaired. Two weeks after IANX, pinching or heat stimulus-induced extracellular signal-regulated kinase phosphorylation (pERK) was restored to the control level, but in the case of pinch stimulation the distribution pattern of pERK-IR cells was altered in the Vc. Taken together, our results support the possibility that peptidergic neurons regenerate more efficiently than non-peptidergic neurons after trigeminal nerve injury.


Pattern sequence recognition using a time-varying Hopfield network.
This paper presents a novel continuous-time Hopfield-type network which is effective for temporal sequence recognition. The fundamental problem of recalling pattern sequences by neural networks is first reviewed. Since it is difficult to implement a desired flow vector field distribution by using conventional matrix encoding scheme, a time-varying Hopfield model (TVHM) is proposed. The weight matrix of the TVHM is constructed in such a way that its auto-correlation and cross-correlation parts are encoded from two different sets of patterns. With this mechanism, flow vectors between any two adjacent stored patterns are of the same directions. Moreover, the flow vector field distribution around a stored pattern can be modulated by the time variable. Then, theoretical results regarding the radii of attraction and the recalling dynamics of the TVHM are presented. The proposed approach is different from the existing methods because neither synchronous dynamics nor interpolated training patterns are required. A way of increasing the storage capacity of the TVHM is proposed. Finally, experimental results are presented to illustrate the validity, capacity, recall capability, and the applications of the proposed model.


Oxygen saturation targeting in preterm infants receiving continuous positive airway pressure.
The precision of oxygen saturation (SpO2) targeting in preterm infants on continuous positive airway pressure (CPAP) is incompletely characterized. We therefore evaluated SpO2 targeting in infants solely receiving CPAP, aiming to describe their SpO2 profile, to document the frequency of prolonged hyperoxia and hypoxia episodes and of fraction of inspired oxygen (FiO2) adjustments, and to explore the relationships with neonatal intensive care unit operational factors. Preterm infants <37 weeks' gestation in 2 neonatal intensive care units were studied if they were receiving CPAP and in supplemental oxygen at the beginning of each 24-hour recording. SpO2, heart rate, and FiO2 were recorded (sampling interval 1-2 seconds). We measured the proportion of time spent in predefined SpO2 ranges, the frequency of prolonged episodes (≥30 seconds) of SpO2 deviation, and the effect of operational factors including nurse-patient ratio. A total of 4034 usable hours of data were recorded from 45 infants of gestation 30 (27-32) weeks (median [IQR]). When requiring supplemental oxygen, infants were in the target SpO2 range (88%-92%) for only 31% (19%-39%) of total recording time, with 48 (6.9-90) episodes per 24 hours of severe hyperoxia (SpO2 ≥98%), and 9.0 (1.6-21) episodes per 24 hours of hypoxia (SpO2 <80%). An increased frequency of prolonged hyperoxia in supplemental oxygen was noted when nurses were each caring for more patients. Adjustments to FiO2 were made 25 (16-41) times per day. SpO2 targeting is challenging in preterm infants receiving CPAP support, with a high proportion of time spent outside the target range and frequent prolonged hypoxic and hyperoxic episodes.


Moringa oleifera-rich diet and T cell calcium signaling in spontaneously hypertensive rats.
Moringa oleifera is a plant whose fruits, roots and leaves have been advocated for traditional medicinal uses. The physicochemical analysis shows that Moringa oleifera contains more dietary polyunsaturated fatty acids (PUFA) than saturated fatty acids (SFA). The consumption of an experimental diet enriched with Moringa oleifera extracts lowered blood pressure in spontaneously hypertensive rats (SHR), but not in normotensive Wistar-Kyoto (WKY) rats as compared to rats fed an unsupplemented control diet. Anti-CD3-stimulated T cell proliferation was diminished in both strains of rats fed the Moringa oleifera. The experimental diet lowered secretion of interleukin-2 in SHR, but not in WKY rats compared with rats fed the control diet. Studies of platelets from patients with primary hypertension and from SHR support the notion that the concentration of intracellular free calcium [Ca(2+)](i) is modified in both clinical and experimental hypertension. We observed that the basal, [Ca(2+)](i) was lower in T cells of SHR than in those of WKY rats fed the control diet. Feeding the diet with Moringa oleifera extracts to WKY rats did not alter basal [Ca(2+)](i) in T cells but increased basal [Ca(2+)](i) in SHR. Our study clearly demonstrated that Moringa oleifera exerts antihypertensive effects by inhibiting the secretion of IL-2 and modulates T cell calcium signaling in hypertensive rats.


Difference in Diffusion-Weighted Magnetic Resonance Imaging and Diffusion Tensor Imaging Parameters Between Endometrioid Endometrial Adenocarcinoma and Uterine Serous Adenocarcinoma: A Retrospective Study.
This study aimed to elucidate the difference in diffusion-weighted magnetic resonance imaging (DWI) and diffusion tensor imaging (DTI) parameters between endometrioid endometrial adenocarcinoma (EEA) and uterine serous adenocarcinoma (SA). Data of patients with pathologically confirmed EEA or SA who underwent DWI and DTI scanning between May 2013 and July 2016 were retrospectively analyzed. Apparent diffusion coefficient (ADC) value from DWI and ADC from DTI (ADC) map and fractional anisotropy (FA) values from DTI were analyzed and compared statistically. The correlation between ADC and ADC was analyzed by Pearson correlation analysis. Apparent diffusion coefficient, ADC, and FA between the 2 groups were compared using independent t test. The effect of ADC, ADC, and FA in distinguishing EEA and SA was evaluated by receiver operator characteristic curve. Thirty-three patients were enrolled into the study, including 13 cases of SA and 20 cases of EEA. Pearson correlation analysis suggested that the value of ADC was highly related with ADC in both the SA group (r = 0.812, P = 0.001) and the EEA group (r = 0.858, P < 0.001). The value of ADC and ADC in the SA group was significantly lower than that in the EEA group; FA was significantly higher than that in the EEA group. Receiver operator characteristic curve analysis showed that ADC and ADC have high sensitivity and specificity; FA has low sensitivity and high specificity. We suggest that both DWI and DTI could be used in distinguishing EEA from SA. Apparent diffusion coefficient and ADC possess potential diagnostic value with high sensitivity and specificity.


Transanal Mesenteric Resection in Hirschsprung's Disease Using ICG under Concept of NOTES Technique.
Laparoscopic surgery has been applied for Hirschsprung's disease (HD). We herein report our approach to mesenteric processing for laparoscopic-assisted transanal endorectal pull-through (L-TERPT). Following mucosectomy and entering the abdominal cavity, a vessel sealing system is transanally inserted into the abdominal cavity for mesenteric processing based on concept of Natural Orifice Translumenal Endoscopic Surgery. Since the transanal axis is parallel to the dissected mesentery, it makes easier to operate in comparison to when the procedure is performed through the abdominal working port and can reduce the additional abdominal trocar wound. We also use indocyanine green (ICG) fluorescence navigation. Fluorescing the vessels with ICG allows intraoperative visualization of the blood flow in the retrieved intestine. With these innovative combined techniques, L-TERPT for HD can be safely performed, even in infants with small intraabdominal cavities.


Evaluation of prostate cancer characteristics in four populations worldwide.
Prostate cancer is common around the world, but rates of advanced disease differ substantially by race and geography. Although a major health issue, little is known about prostate cancer presentation in West Africa and India compared to the United States (US). The aim of this study was to compare prostate tumor characteristics in four populations of men from the US, Senegal and India. We recruited prostate cancer patients from four hospital-based populations. The sample included 338 African-Americans, 1265 European-Americans, 122 Asian Indians, and 72 Senegalese. Questionnaire and medical record data were collected on each participant. We found significant differences in age at diagnosis, BMI, and PSA levels across the groups. Senegalese and Indian men had a higher probability of high stage (T3/T4) disease compared to US men. Gleason grade was significantly higher in Asian Indians compared to other populations. African-Americans, Senegalese, and Asian Indians had a significantly higher probability of metastatic disease compared to European Americans. The odds ratios (OR) for metastasis were consistently higher in Asian Indians compared to American cases. As only 19/72 Senegalese were assessed for metastasis, OR could not be determined for metastasis. These results suggest that there are significant geographical and ethnic differences in the presentation of prostate cancer. Men in developing countries tend to present with advanced disease compared to US men. Identifying risk factors for advanced disease may help to decrease the rate of poor prostate cancer outcomes and associated mortality worldwide.


Evaluation of attendance and weight loss in an intensive weight management clinic compared to standard dietetic care.
With increasing rates of obesity, the effectiveness of weight reduction programmes come under increasing scrutiny. This nonrandomized study aimed to review the effectiveness of two weight loss clinics in terms of percent and rate of weight loss, and attendance. Data were collected on consecutive obese patients, attending either an intensive weight management clinic (IWMC) or a general dietetic outpatient clinic. The IWMC had a structured approach with six once-a-month appointments, a signed agreement to attend, an initial screening of readiness to change and consistent advice from one dietitian. The general clinic was less structured, had more ad hoc follow up and did not guarantee one dietitian. Seventy percent of patients referred were female [mean (SD) age 48 (14.2) years]. Thirty-three percent (103/313) of all patients referred did not book an appointment. Of those attending with a body mass index > or = 32 kg m(-2), 55% were seen in the general and 45% in the intensive clinic, but only 19% and 53%, respectively, completed the programmes. The total amount and rate of weight loss did not differ significantly between clinics. However, analysis using the last recorded weight revealed a median weight reduction of 1.8% (interquartile range = -5.6-0) at the median rate of -0.4 kg month(-1) (-1-0) in the intensive clinic, compared to no overall weight loss in the general clinic (P < or = 0.001). A more structured approach and initial screening of readiness to change is likely to achieve better weight loss results and therefore will comprise a better use of dietetic time than including obese patients in general clinics.


No evidence that selection for egg production persistency causes loss of bone quality in laying hens.
The physiological adaptations that have evolved for egg laying make hens susceptible to bone fractures and keel bone damage. In modern laying hen breeds, longer periods of egg laying could result in a greater risk of poor bone quality, and selection for increased egg production has frequently been stated to be a cause. However, the existing literature does not support this hypothesis. To test the hypothesis that egg production is associated with quality, breaking strength and density of bone, genetic correlations between these traits were estimated in White Leghorn and Rhode Island Red breeds. Genetic correlations of cortical and medullary bone material chemical properties with bone quality were also estimated, in order to identify methods to improve bone quality with appropriately targeted measurement of key traits. Estimates of heritability for bone quality traits were moderate (0.19-0.59) for both White Leghorn and Rhode Island Red breeds, except for the keel bone trait, which had a heritability estimate equal to zero. There was no evidence for genetic or phenotypic relationships between post-peak egg production and bone quality. In the White Leghorn breed, the estimate of the genetic correlation between pre-peak production/age at first egg and bone quality was significant and negative (- 0.7 to - 0.4). Estimates of heritability of thermogravimetric measurements of tibial medullary bone mineralisation were significant (0.18-0.41), as were estimates of their genetic correlations with tibia breaking strength and density (0.6-0.9). The low genetic correlation of post-peak egg production with bone quality suggests that selection for increased persistency of egg production may not adversely affect bone quality. Onset of puberty and mineralisation of the medullary bone, which is a specialised adaptation for egg laying, were identified as important factors associated with the quality of the skeleton later during egg production. These are traits for which genetic, as well as environmental and management factors can positively impact the overall quality of the skeleton of laying hens.


Adsorption of HMF from water/DMSO solutions onto hydrophobic zeolites: experiment and simulation.
The adsorption of 5-hydroxymethylfurfural (HMF), DMSO, and water from binary and ternary mixtures in hydrophobic silicalite-1 and dealuminated Y (DAY) zeolites at ambient conditions was studied by experiments and molecular modeling. HMF and DMSO adsorption isotherms were measured and compared to those calculated using a combination of grand canonical Monte Carlo and expanded ensemble (GCMC-EE) simulations. A method based on GCMC-EE simulations for dilute solutions combined with the Redlich-Kister (RK) expansion (GCMC-EE-RK) is introduced to calculate the isotherms over a wide range of concentrations. The simulations, using literature force fields, are in reasonable agreement with experimental data. In HMF/water binary mixtures, large-pore hydrophobic zeolites are much more effective for HMF adsorption but less selective because large pores allow water adsorption because of H2 O-HMF attraction. In ternary HMF/DMSO/water mixtures, HMF loading decreases with increasing DMSO fraction, rendering the separation of HMF from water/DMSO mixtures by adsorption difficult. The ratio of the energetic interaction in the zeolite to the solvation free energy is a key factor in controlling separation from liquid mixtures. Overall, our findings could have an impact on the separation and catalytic conversion of HMF and the rational design of nanoporous adsorbents for liquid-phase separations in biomass processing.


Serum basal tryptase levels in healthy children: correlation between age and gender.
Tryptase is used as a marker in many clinical conditions such as mast cell activation and systemic anaphylaxis. Normal levels of the serum basal tryptase (sBT) are determined in adulthood; however, data about nonallergic healthy children is limited. This study was designed to determine the normal sBT levels in healthy children. Total sBT levels were measured in 113 sera from healthy nonallergic children, for routine follow-up or diagnosis of illnesses that are not known to induce changes in serum tryptase levels. One hundred thirteen children aged 0.16-9.91 years (male/female subjects, 68/45 [60%/40%]) with a median (interquartile) age of 3.36 years (1.94-5.68 years) were evaluated. The sBT level was determined to be 3.30 ng/mL (2.38-4.36) median (interquartile) for the whole group. There was a tendency of higher sBT levels for boys than girls (3.49 [2.56-4.64 ng/mL] and 2.91 ng/mL [2.16-4.16 ng/mL], respectively); however, these findings were not significant (p = 0.12). The analysis of the whole group revealed that sBT levels were inversely related with age (r = -0.259; p = 0.006); and the decrease of sBT with age was more prominent in girls (r = -0.282; p = 0.02) than in boys. The highest sBT levels were found in the 0- to 1-year age group (all, 4.67 ng/mL [4.04-6.39 ng/mL]; boys, 5.34 ng/mL [4.04-6.39 ng/mL]; girls, 4.48 ng/mL [3.23-16.26 ng/mL]). The sBT levels in healthy children are similar to those in adults except in infancy. Interestingly, sBT levels tend to decrease slightly with age. Although they did not reach significance, sBT levels were found to be higher in boys compared with the girls.


Bending of conjugated molecular wires and its effect on electron conduction properties.
The electronic structure and electron transport properties of simple conjugated molecular wires like oligophenylene ethynylene (OPE) and oligophenylene vinylene (OPV) are studied under compression. If artificially confined to a given shorter length, the oligomers tend to bend and bending causes a loss in the overlap of the conjugated molecular orbitals. Theoretical modeling of electronic transport has been carried out for all undistorted and compressed OPE/OPV oligomers. OPV exists in step-like or V-like conformations and they have the same stability with very similar frontier molecular orbitals. The conductances of these molecular wires are calculated when inserted between two gold probes and the conductances for OPV are found to be comparable to OPE when the interfaces are same. The conductance decreases with bending due to the gradual loss in overlap of the molecular orbitals. It is also found that the conductances of the molecular wires decrease very strongly if the terminal sulfur atom is simultaneously bonded to hydrogen and a gold surface, thus reflecting the importance of the interface in determining the conductance in two-probe systems. From the conductance studies it may be concluded that if one or more benzene rings of OPE are rotated from coplanar conditions, the orthogonal molecular orbitals may completely block the electronic transport, rendering the molecule insulating.


A rapidly produced 125I labelled autologous fibrinogen: in vitro properties and preliminary metabolic studies in man.
The properties of fibrinogen extracted by a precipitation method using glycine at ambient temperatures near neutral pH are described. The simple and reproducible method gives a 73% yield of high purity plasminogen-free fibrinogen in 45 minutes from small volumes of plasma. The protein extract was labelled with 125I using chloramine-T under conditions optimal for fibrinogen stability. The extraction procedure, radio-iodination, desalting, and sterilization take only 70 minutes for completion from the time donor blood is received in the laboratory. The methods, using a specially developed extraction vessel and desalting/sterilizing column, can be used in a small hospital laboratory. Autologous fibrinogen can thus be extracted from patients' blood, eliminating the risk of transmitting hepatitis when it is re-administered. The autologous material, which is 97% clottable and contains less than 0-05% free iodide, is being routinely used as a diagnostic tool in the detection of deep vein thrombosis. The high purity of the preparation facilitates metabolic studies and in vitro experimental work. In vivo results show a mean half-life in three normal volunteers of 3-95 days and a catabolic rate of 25-23% per day with the extravascular space estimated as 24-86%. In 30 surgical patients an expected reduced half-life in plasma was determined with a mean of 3-1 days.


Functional, bioactive and antigenicity properties of blue whiting protein hydrolysates: effect of enzymatic treatment and degree of hydrolysis.
Fish discards represent an important under-utilisation of marine resources. This study evaluated the up-grading of the protein fraction of blue whiting (Micromesistius poutassou) discards by the production of fish protein hydrolysates (FPHs) exhibiting functional, antioxidant, angiotensin-I converting enzyme (ACE)-inhibitory and antigenicity properties. FPHs with low DH (4%) showed better emulsifying, foaming and oil binding capacities, particularly those obtained using only trypsin. FPHs with DH 4% exhibited also the stronger antioxidant activity, especially the one obtained using only subtilisin (IC50 = 1.36 mg protein mL-1 ). The presence of hydrophobic residues at the C-terminal of the FPH produced using subtilisin also led to the stronger ACE-inhibitory activity. However, FPHs with high DH (12%), which implies a higher proportion of short peptides, was required to enhance ACE-inhibition (IC50 = 172 µg protein mL-1 ). The antigenic levels of the FPH were also reduced with DH independently of the enzymatic treatment. Nevertheless, the highest degradation of fish allergens (e.g. parvalbumin) was also obtained when using only subtilisin. These results suggest that added-value products for food applications can be produced from the protein fraction of discards. © 2016 Society of Chemical Industry.


Forecasting deforestation and carbon emissions in tropical developing countries facing demographic expansion: a case study in Madagascar.
Anthropogenic deforestation in tropical countries is responsible for a significant part of global carbon dioxide emissions in the atmosphere. To plan efficient climate change mitigation programs (such as REDD+, Reducing Emissions from Deforestation and forest Degradation), reliable forecasts of deforestation and carbon dioxide emissions are necessary. Although population density has been recognized as a key factor in tropical deforestation, current methods of prediction do not allow the population explosion that is occurring in many tropical developing countries to be taken into account. Here, we propose an innovative approach using novel computational and statistical tools, including R/GRASS scripts and the new phcfM R package, to model the intensity and location of deforestation including the effect of population density. We used the model to forecast anthropogenic deforestation and carbon dioxide emissions in five large study areas in the humid and spiny-dry forests of Madagascar. Using our approach, we were able to demonstrate that the current rapid population growth in Madagascar (+3.39% per year) will significantly increase the intensity of deforestation by 2030 (up to +1.17% per year in densely populated areas). We estimated the carbon dioxide emissions associated with the loss of aboveground biomass to be of 2.24 and 0.26 tons per hectare and per year in the humid and spiny-dry forest, respectively. Our models showed better predictive ability than previous deforestation models (the figure of merit ranged from 10 to 23). We recommend this approach to reduce the uncertainty associated with deforestation forecasts. We also underline the risk of an increase in the speed of deforestation in the short term in tropical developing countries undergoing rapid population expansion.


Expressional changes of genes and miRNA in common megakaryocyte-erythroid progenitors from lower-risk myelodysplastic syndrome.
Myelodysplastic syndrome (MDS) is a stem cell tumor characterized by dysplastic features and ineffective hematopoiesis in the early phase and leukemic progression in the late phase. Speculating that differences in the expression of genes and microRNA (miRNA) in control and MDS-derived erythroid progenitors may cause ineffective erythropoiesis, we sorted common megakaryocyte-erythroid progenitors (MEPs) in bone marrow cells from three lower-risk MDS patients, and compared expression levels of genes and miRNA with those from controls. In apoptosis-related pathways, the expression of some pro-apoptotic genes, such as cell death-inducing DFFA-like effector A, caspase 5, and Fas ligand, was elevated in MDS-derived MEPs, while those of anti-apoptotic CD40 and tumor necrosis factor were lower. In hematopoiesis-regulating pathways, RUNX1 and ETV6 genes showed reduced expression. Expression profiling revealed that three and 35 miRNAs were significantly up- and down-regulated in MDS-derived MEPs. MIR9 exhibited robust expression in MEPs and CD71+GlyA+ erythroid cells derived from one of the three patients. Interestingly, overexpression of MIR9 inhibited the accumulation of hemoglobin in UT-7/GM cells. Some of these alterations in gene and miRNA expression may contribute to the pathogenesis of ineffective hematopoiesis in lower-risk MDS and provide molecular markers for sub-classification and making a prognosis.


Aging restricts the ability of mesenchymal stem cells to promote the generation of oligodendrocytes during remyelination.
Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) that leads to severe neurological deficits. Due to their immunomodulatory and neuroprotective activities and their ability to promote the generation of oligodendrocytes, mesenchymal stem cells (MSCs) are currently being developed for autologous cell therapy in MS. As aging reduces the regenerative capacity of all tissues, it is of relevance to investigate whether MSCs retain their pro-oligodendrogenic activity with increasing age. We demonstrate that MSCs derived from aged rats have a reduced capacity to induce oligodendrocyte differentiation of adult CNS stem/progenitor cells. Aging also abolished the ability of MSCs to enhance the generation of myelin-like sheaths in demyelinated cerebellar slice cultures. Finally, in a rat model for CNS demyelination, aging suppressed the capability of systemically transplanted MSCs to boost oligodendrocyte progenitor cell (OPC) differentiation during remyelination. Thus, aging restricts the ability of MSCs to support the generation of oligodendrocytes and consequently inhibits their capacity to enhance the generation of myelin-like sheaths. These findings may impact on the design of therapies using autologous MSCs in older MS patients.


Relationship between the small cervical vertebral body and the morbidity of cervical spondylosis.
This study aimed to determine the relationship between the size of the cervical vertebral body and the morbidity of cervical spondylosis, and to examine the characteristics of spondylosis patients with small cervical vertebral bodies.The clinical data and the sagittal reconstructions of computed tomography images of 182 patients with cervical spondylosis were collected retrospectively. Patients included 74 males and 108 females, with a mean age of 31.8 years (range 20-40 years). The Torg-Pavlov ratio and the sagittal diameter of the vertebral body were measured. A Torg-Pavlov ratio above 1.2 was regarded as a small cervical vertebral body (SCVB), and below 1.2 as a nonsmall vertebral body (NSCVB).The NSCVB group was more prone to neurological symptoms than was the SCVB group (P < .05). There was no significant difference in neck pain between the 2 groups (P > .05). Conservative treatment achieved similar recovery rates in the SCVB group and the NSCVB group (81.8% vs 93.6%; P > .05). The rate of symptom (eg, axial neck pain) recurrence and persistence in the SCVB group was significantly higher than in the NSCVB group (P < .05).Our study found that smaller size of the cervical vertebral body is an attributing factor for cervical spondylosis. Patients with smaller cervical vertebral bodies are prone to persistent axial neck pain, but not neurological symptoms.


Modeling ozone removal to indoor materials, including the effects of porosity, pore diameter, and thickness.
We develop an ozone transport and reaction model to determine reaction probabilities and assess the importance of physical properties such as porosity, pore diameter, and material thickness on reactive uptake of ozone to five materials. The one-dimensional model accounts for molecular diffusion from bulk air to the air-material interface, reaction at the interface, and diffusive transport and reaction through material pore volumes. Material-ozone reaction probabilities that account for internal transport and internal pore area, γ(ipa), are determined by a minimization of residuals between predicted and experimentally derived ozone concentrations. Values of γ(ipa) are generally less than effective reaction probabilities (γ(eff)) determined previously, likely because of the inclusion of diffusion into substrates and reaction with internal surface area (rather than the use of the horizontally projected external material areas). Estimates of γ(ipa) average 1 × 10(-7), 2 × 10(-7), 4 × 10(-5), 2 × 10(-5), and 4 × 10(-7) for two types of cellulose paper, pervious pavement, Portland cement concrete, and an activated carbon cloth, respectively. The transport and reaction model developed here accounts for observed differences in ozone removal to varying thicknesses of the cellulose paper, and estimates a near constant γ(ipa) as material thickness increases from 0.02 to 0.16 cm.


Essential fatty acids in health and disease.
Essential fatty acids (EFAs) form an important component of cell membranes, are eicosanoid precursors and are therefore required for both the structure and function of every cell in the body. EFAs can modulate the activity of protein kinase C, T and B cell response, free radical generation and lipid peroxidation, lymphokine secretion and cell proliferation. EFAs also have anti-mutagenic, anti-bacterial, anti-fungal and anti-viral properties. EFAs and their metabolites lower serum cholesterol, triglycerides and blood pressure. EFAs appear to be of benefit in atopic eczema, premenstrual syndrome, psoriasis, auto-immune disorders especially rheumatoid arthritis and systemic lupus erythematosus, prevention of target organ damage in diabetes mellitus, peptic ulcer disease, ulcerative colitis, coronary heart disease and atherosclerosis. EFAs and their metabolites can selectively kill tumor cells both in vitro and in vivo without harming normal cells. In addition, EFAs seem to play a fundamental role in inflammation and immune response. In view of their actions and relative safety, it is anticipated that EFAs may be useful in the management of several diseases.


Treatment and outcomes of patients with primary breast sarcoma.
Surgery is the main treatment for primary breast sarcoma (PBS). Here we characterize this disease and determine factors associated with use of adjuvant therapy. Records of patients with PBS from 1986 to 2006 were reviewed. Overall survival (OS) was estimated by Kaplan-Meier. Relationships between patient variables and OS were determined using univariate Cox proportional hazard models. Thirteen patients with PBS were identified; 10 patients underwent mastectomy, and 3 underwent partial mastectomy. Six patients underwent axillary staging; none were positive. Patients with tumors >5 cm were more likely to undergo radiation therapy (P <.05). Local recurrence occurred in 7 patients. Metastatic disease was present in 2 patients at diagnosis, and 6 patients developed metastatic disease; all 8 patients died from their disease. Five patients remained disease free. Five-year OS was 67% (83% for tumors <5 cm and 42% for tumors >5 cm). Tumor size was significantly associated with OS (relative risk = 1.1/1 cm increase in size > 5 cm). Treatment for PBS is excision to clear margins. Axillary staging is not indicated. Tumor size >5 cm is the only significant prognostic indicator of overall survival.


[The use of the direct basophil degranulation test for the diagnosis of allergy].
Basophil degranulation test (BDT) was used in examinations of patients suffering from allergies. A modified technique was employed with toluidine blue staining and basophil scintillation in a counter chamber. The examinees were divided into three groups. Group 1 (n = 34) consisted of patients suffering from atopic bronchial asthma with suspected sensitization to home dust. Group 2 included 118 patients with a history of allergy to penicillin. Group 3 consisted of 51 patients with drug allergies (streptomycin, vitamin B1, procaine, analgin intolerance). In all the cases specific diagnosis was confirmed by skin and provocation test. 71.4 percent of the patients with sensitization to home dust allergen showed positive results in BDT, as did 68.6 percent of Group 3 subjects. Of the cases with penicillin sensitization confirmed by application test 91.7 percent were positive in BDT, of those with positive intradermal tests, 66.7 percent were BDT-positive, this being probably explained by individual sensitivity of the patients. These results recommend BDT, an informative and simple technique, for wide practice.


[Influence of ionizing radiation on the parameters of oxidative stress markers in the rat liver and thymus].
Influence of ionizing radiation on the parameters of oxidative stress markers in the liver and thymus of the rats exposed to gamma-radiation 60Co at a doze of 4 Gy was investigated. The animals were decapitated on the 1, 4, 7, 10, 14, 22 and 30th day after irradiation and cell suspensions from the liver and thymus were obtained. After centrifugation, the content of MDA, the spontaneous and NADH-induced synthesis of superoxide anion radical of oxygen, the content of total and free iron were determined in the cellular sediment and centrifugate containing intercellular fluid. It is shown that the content of MDA and the levels of spontaneous and NADH-induced synthesis of superoxide anion radical of oxygen increases in intercellular fluid and thymus and liver cells on the 1st day after radiation exposure. In the liver, these parameters are normalized by the 4th day and do not significantly differ from the control level in the period of time following radiation exposure. In thymus, as compared with liver, the level of oxidative stress parameters increases by the 4th day after radiation and remains at the raised level within 22 days after irradiation exposure. It is shown that the content of free iron in thymus cells of irradiated animals increases 3.6 times by the 4th day and reliably exceeds the control level within the next 22 days. Radiation does not lead to any changes in the content of free iron in liver cells. Different levels of the free iron content can serve the reason for various sensitivity of oxidative stress markers in thymus and liver cells to radiation exposure.


Evaluation of the serum level of estrogen, progesterone, prolactin, and testosterone in patients with trigeminal neuralgia compared to a healthy population.
The goal of this study is to measure and compare the hormonal serum levels (estrogen, progesterone, testosterone, prolactin, dihydrotestosterone [DHT]) in trigeminal neuroglia (TN) menopausal women and healthy women. This cross-sectional and case-control study was performed in 2018 and 2019. For this study, menopausal women with confirmed TN were enrolled. Twenty-two healthy women in the control group and 19 in the case group participated. Blood samples were taken from participants for assessment of hormonal serum levels (estrogen, progesterone, testosterone, prolactin, DHT). Data were analyzed by SPSS version 18. Mann-Whitney, T-test, kormography test, nonmetric, χ2 test, and odds ratios have been used. In patients with TN, the serum level of testosterone was significantly higher (p = .036), and the serum level of prolactin (p = .016) was significantly lower. Other evaluated hormones' serum level was identical in the two groups. Patients with abnormal estrogen levels were more in the TN group in comparison with the healthy group. The abnormality of progesterone in TN patients was more in comparison to the healthy control group. Estrogen and progesterone serum levels in TN patients are higher in comparison with the healthy group, while prolactin and testosterone serum levels are lower in the control group. Moreover, the DHE serum level is similar in both groups.


Combinational application of metal-organic frameworks-based nanozyme and nucleic acid delivery in cancer therapy.
The rapid development of nanotechnology has generated numerous ideas for cancer treatment, and a wide variety of relevant nanoparticle platforms have been reported. Metal-organic frameworks (MOFs) have been widely investigated as an anti-cancer drug delivery vehicle owing to their unique porous hybrid structure, biocompatibility, structural tunability, and multi-functionality. MOF materials with catalytic activity, known as nanozymes, have applications in photodynamic and chemodynamic therapy. Nucleic acids have also attracted increasing research attention owing to their programmability, ease of synthesis, and versatility. A variety of functional DNAs and RNAs have been applied both therapeutically (gene-targeting drugs for cancer treatment) and nontherapeutically (used as modified materials to enhance the therapeutic effects of other nanomedicines). The combined use of MOFs and functional nucleic acids have been extensively investigated and has been associated with excellent tumor-suppressor activity in various treatment methods. In this review, we summarize the progress in the research and development of tumor therapy based on MOFs and nucleic acid delivery over recent years, focusing on the combinational use of different delivery and design strategies for MOF/therapeutic nucleic acid platforms. We further summarize the strategies for combining MOFs (universal carrier, functional carrier) and nucleic acids (therapeutic nucleic acids, nontherapeutic nucleic acids) and discuss the corresponding therapeutic effects in cancer treatment. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology Therapeutic Approaches and Drug Discovery > Emerging Technologies.


Hexaminolevulinate Blue-Light Cystoscopy in a Patient with Metastatic Melanoma of the Bladder.
Although bladder cancer is one of the most frequently diagnosed tumors worldwide, metastatic melanoma of the bladder is a rare occurrence with only 29 cases reported in the literature. We present the case of a 60-year-old male with a medical history significant for metastatic melanoma, who was referred to the urology department for gross hematuria. Transurethral resection of bladder tumor (TURBT) was performed with the assistance of hexaminolevulinate acid (HAL) with blue-light cystoscopy (BLC). Subsequent histopathologic analysis of the specimen confirmed a diagnosis of metastatic melanoma of the bladder. To our knowledge, this is the first reported case of metastatic bladder melanoma diagnosed with the assistance of HAL-BLC in a patient undergoing a TURBT. Although HAL-BLC is only indicated for use in the cystoscopic detection of papillary nonmuscle invasive bladder cancer, it may aid in the detection of nonconventional bladder pathologies, such as melanoma.


Structural analysis of the major immediate early gene of human cytomegalovirus.
The most abundant species of human cytomegalovirus (Towne) immediate early polysome-associated RNA originates from a region of ca. 2.8 kilobases (0.739 to 0.755 map units) within the XbaI-E DNA fragment. These sequences code for a 1.95-kilobase mRNA and are referred to as immediate early coding region one (M. F. Stinski, D. R. Thomsen, R. M. Stenberg, and L. C. Goldstein, J. Virol. 46:1-14, 1983). We have utilized the nuclease mapping technique of Berk and Sharp (A. J. Berk and P. A. Sharp, Cell 12:721-732, 1977) to examine this gene in detail. Cloned fragments of human cytomegalovirus DNA, either labeled with 32P in vivo or end labeled in vitro at the 5' or 3' termini, were hybridized to immediate early polysome-associated RNA. The hybrids were treated with single-strand-specific nuclease and subjected to electrophoresis in either neutral or denaturing gels. The major transcript was shown to be a spliced molecule containing a 3' terminal exon of 1,341 nucleotides. Upstream of the major body of the mRNA are three small exon sequences of 185, 88, and 121 nucleotides. The sequence of the exons as well as the locations of the intron-exon splice junctions were determined. Based on the DNA sequence, the viral mRNA molecule has one open reading frame which begins within the second exon and extends for 491 amino acid residues. The predicted molecular weight of the polypeptide originating from this region was estimated to be 64,000. It is hypothesized that this viral gene codes for the major regulatory protein controlling transcription of the viral genome at early times. The properties of the viral gene and its protein product are discussed.


Shoulder injuries in the athlete.
Most shoulder injuries in the athlete can be managed nonoperatively; prior to any management, however, they require careful evaluation for accurate diagnosis. Once the diagnosis has been made, few of these injuries will actually require surgery and most will obtain an excellent result with full return to function. Several specific shoulder injuries are discussed and their treatment is outlined.


Bacterial Cooperation Causes Systematic Errors in Pathogen Risk Assessment due to the Failure of the Independent Action Hypothesis.
The Independent Action Hypothesis (IAH) states that pathogenic individuals (cells, spores, virus particles etc.) behave independently of each other, so that each has an independent probability of causing systemic infection or death. The IAH is not just of basic scientific interest; it forms the basis of our current estimates of infectious disease risk in humans. Despite the important role of the IAH in managing disease interventions for food and water-borne pathogens, experimental support for the IAH in bacterial pathogens is indirect at best. Moreover since the IAH was first proposed, cooperative behaviors have been discovered in a wide range of microorganisms, including many pathogens. A fundamental principle of cooperation is that the fitness of individuals is affected by the presence and behaviors of others, which is contrary to the assumption of independent action. In this paper, we test the IAH in Bacillus thuringiensis (B.t), a widely occurring insect pathogen that releases toxins that benefit others in the inoculum, infecting the diamondback moth, Plutella xylostella. By experimentally separating B.t. spores from their toxins, we demonstrate that the IAH fails because there is an interaction between toxin and spore effects on mortality, where the toxin effect is synergistic and cannot be accommodated by independence assumptions. Finally, we show that applying recommended IAH dose-response models to high dose data leads to systematic overestimation of mortality risks at low doses, due to the presence of synergistic pathogen interactions. Our results show that cooperative secretions can easily invalidate the IAH, and that such mechanistic details should be incorporated into pathogen risk analysis.


What do people with MS want and expect from health-care services?
To determine the health-care preferences of people with Multiple Sclerosis (MS). Cross-sectional survey using a postal questionnaire comprising standardized measures of health related quality of life (SF-36) and of depression (BDI) and original questions about issues linked to health-care, prioritized by people with MS during an earlier qualitative phase. A stratified sample of 318 people with MS in Scotland and England. Questionnaires were sent to 471 people with multiple sclerosis. The 318 respondents (68%) reported a wide variation in quality of life, but on average scored significantly lower than normative values in all dimensions of the SF-36. The most commonly used home treatment during the previous year was evening primrose oil, taken by almost half of the sample (47%). Use of cannabis was acknowledged by 8%. There was evidence that many preferences concerning health services were not satisfied. For instance lack of advice about at least one MS related issue was reported lacking for three-quarters of the sample and 17% lacked advice in at least five areas. Advice about exercise was the single most requested area. The most frequently consulted health professional was the GP (78%) followed by the hospital consultant (50%). The professional that respondents most commonly said they would like to have seen was a specialist MS nurse (30%). Forty-three percent of the sample said they had attended a special meeting for people with MS or disabilities. They were equally divided between those who did and who did not find their last attendance helpful. People with MS display a wide variation in their preference for services and unmet needs. Information about management (both conventional and unconventional), relevant tailored advice and access to appropriately skilled professionals should be feasible components of high quality care. This work has highlighted the value of involving people with MS in the identification of their preferences; further research is needed to show how these might be provided most effectively.


Myxedema.
This review will discuss generalized myxedema as it develops in hypothyroidism. First, the precipitating conditions (thyroprivic trophoprivic + goitrous forms) and the clinical manifestations of thyroid hormone deficiency are presented. Pathobiochemical and pathophysiological factors that lead to the main manifestations include retention of fluid, retention of sodium and hyponatremia. In particular are primary and direct consequences of reduced thyroid hormone levels, and secondary or indirect consequences, such as cardiovascular and renal derangements. In hypothyroidism many biochemical disturbances result. Most important is the interstitial deposition of hydrophilic mucopolysaccharides, which in turn lead to fluid and Na retention and impairment of blood circulation and lymphatic drainage. Myxedema, therefore, is to a large extent a lymphatic edema. Hyponatremia is an indirect consequence of the lack of T3 and is directly caused by impaired renal Na reabsorption. Renal Na,K-ATPase is reduced in specific segments. The often discussed role of inappropriate elevation of circulating ADH does not seem to be a key factor in myxedema. Impaired capacity of renal water excretion is caused by reduced GFR. We discuss the time dependent development of the derangement of different organ systems, and include recently published biochemical results, according to which the lack of T3 interferes not only with the metabolism of numerous compounds of the interstitial matrix, but also with cell surface proteins and intracellular proteins of microfilaments. Finally, we refer briefly to pretibial myxedema in states of hyperthyroidism, that is, infiltrative dermopathy in Graves' disease, which is caused by poorly understood autoimmune processes.


Rhino-orbital-cerebral zygomycosis in solid organ transplant recipients.
Rhino-orbital-cerebral disease is a significant manifestation of zygomycosis in solid organ transplant (SOT) recipients. However, its characteristics and outcome are not well addressed. SOT recipients with zygomycosis as per the European Organization for Research and Treatment in Cancer and the Mycoses Study Group criteria in a cohort study at our centers published previously and those identified with a PubMed search from the 1950s to November 2009 were studied. Patients with mycosis involving the sinuses, orbits, or central nervous system (CNS) were included. Patients comprised a total of 90 SOT recipients with rhino-orbital-cerebral zygomycosis, including 13 in our cohort and 77 in the literature. CNS disease occurred in 57% (51 of 90). Overall mortality was 52.3% (46 of 88), and the mortality in patients with CNS disease was 73.5% (36 of 49). In logistic regression analysis, older age (odds ratio [OR] 1.12, 95% confidence interval [CI] 1.04-1.21, P=0.002) was associated with a higher mortality rate, whereas lipid formulations of amphotericin B compared with amphotericin B deoxycholate (OR 0.09, 95% CI 0.02-0.50, P=0.006) and surgery (OR 0.12, 95% CI 0.01-0.94, P=0.043) were independently associated with an improved survival even when controlled for CNS involvement and the era of diagnosis of disease. Rhino-orbital-cerebral zygomycosis, particularly CNS disease, is associated with substantial mortality rate in SOT recipients. Older age is a significant risk factor for mortality, whereas lipid formulations of amphotericin B and surgery improved outcomes.


Sleep quality during alcohol withdrawal with bright light therapy.
1. Alcohol withdrawal is a complex syndrome that ranges from anxiety, insomnia to delirium tremens. Common treatment is the application of sedative medication. Exposure to bright light in the daytime should advance the normal sleep/wake cycle and moreover it should improve the availability of man's adaptive behavior during alcohol withdrawal. 2. This pilot study describes bright light therapy (BL) during alcohol withdrawal in ten alcohol dependent patients (DSM-III-R: 291.80) without any sedative medication. BL (3000 Lux) was administered on day 3 of abstinence between 7.00-9.00 a.m. and 5.00-9.00 p.m. Total-sleep-polysomnography (recordings between 10.30 p.m.-6.00 a.m.) and self-rating scale were performed to compare intraindividual changes during three nights. After one adaptation night (immediately after alcohol withdrawal), one baseline night and one "BL-night" and one "post-BL night" were analysed. 3. At baseline, total sleep time and sleep efficiency were severely deteriorated, but tended to improve in the following nights after BL. Sleep onset latency showed a significant decline after BL. Stages 3 and 4 were reduced at baseline. Latencies to slow wave sleep were significantly shortened after BL. REM increased in the nights after BL. Subjective sleep quality improved after BL. Although the present results, bright light having a possible stabilizing effect on sleep maintenance and sleep architecture during acute alcohol withdrawal, the authors could only derive hypotheses for further ongoing controlled investigations using placebo light, to receive final verification.


Cine-CMR partial voxel segmentation demonstrates increased aortic stiffness among patients with Marfan syndrome.
Standard cine-cardiac magnetic resonance (CMR) imaging is commonly used to evaluate cardiac structure, geometry and function. Prior studies have shown that automated segmentation via partial voxel interpolation (PVI) accurately quantifies phantom-based cardiac chamber volumes and necropsy left ventricular myocardial mass. Despite this, the applicability and usefulness of PVI in the determination of physiologic parameters of the aorta such as aortic stiffness has yet to be investigated. Routine CMR was conducted with a 1.5T (GE) scanner with pulse sequences similar to that of standard CMR (parameters: TR 3.4 msec, TE 1.14 msec, flip angle 60°, temporal resolution ~30-40 msec). Views were obtained in standard cardiac-oriented longitudinal or axial views (2, 3 and 4 chambers). Within non-dilated regions of the descending thoracic aorta, aortic area was quantified via a novel PVI automated process (LV-METRIC), which discerns relative amounts of blood pool in each voxel. Aortic stiffness, as calculated from brachial artery pulse pressure and aortic area at maximal and minimal dimensions, was evaluated in 60 total segments (one segment per patient). All segments were in the descending aorta and were not aneurysmal. Sixty patients in total were studied, including 50 that had genetically-related aortic disorder [35 bicuspid aortic valve (BAV), 15 Marfan syndrome (MFS)]. Ten normal controls without aortic disease were included for comparison purposes. All patients (n=60) had evaluable CMR images for assessment of the descending aorta with use of automated segmentation. Patients with BAV and MFS were similar to controls in age, systolic blood pressure, brachial artery pulse pressure, smoking status or hypercholesterolemia (all P=NS). There were more women (P<0.001), lower body mass index (P=0.008), and greater height (P<0.001) in the MFS cohort compared to BAV and controls. Descending aortic area in either systole (maximal) or diastole (minimal) was similar among all three cohorts. However, change in aortic area (ΔArea) throughout the cardiac cycle was substantially lower in MFS than control subjects (P<0.001). In contrast, change in aortic area throughout the cardiac cycle was not significantly different between BAV vs. controls (P=0.62). Aortic stiffness was increased among MFS patients versus control subjects (P=0.014). When comparing MFS to BAV subjects, a comparable trend was observed (P=0.09). No statistical difference was evident in aortic stiffness in patients with BAV versus control subjects (P=0.29). The application of PVI to standard CMR imaging can assess abnormal descending aorta functional indices in normal caliber segments in MFS subjects. Future prospective studies with larger subject populations are warranted to further determine the overall utility of automated aortic segmentation as a possible early biomarker of aortic dysfunction before overt dilatation.


Gas6 Promotes Oligodendrogenesis and Myelination in the Adult Central Nervous System and After Lysolecithin-Induced Demyelination.
A key aim of therapy for multiple sclerosis (MS) is to promote the regeneration of oligodendrocytes and remyelination in the central nervous system (CNS). The present study provides evidence that the vitamin K-dependent protein growth arrest specific 6 (Gas6) promotes such repair in in vitro cultures of mouse optic nerve and cerebellum. We first determined expression of Gas6 and TAM (Tyro3, Axl, Mer) receptors in the mouse CNS, with all three TAM receptors increasing in expression through postnatal development, reaching maximal levels in the adult. Treatment of cultured mouse optic nerves with Gas6 resulted in significant increases in oligodendrocyte numbers as well as expression of myelin basic protein (MBP). Gas6 stimulation also resulted in activation of STAT3 in optic nerves as well as downregulation of multiple genes involved in MS development, including matrix metalloproteinase-9 (MMP9), which may decrease the integrity of the blood-brain barrier and is found upregulated in MS lesions. The cytoprotective effects of Gas6 were examined in in vitro mouse cerebellar slice cultures, where lysolecithin was used to induce demyelination. Cotreatment of cerebellar slices with Gas6 significantly attenuated demyelination as determined by MBP immunostaining, and Gas6 activated Tyro3 receptor through its phosphorylation. In conclusion, these results demonstrate that Gas6/TAM signaling stimulates the generation of oligodendrocytes and increased myelin production via Tyro3 receptor in the adult CNS, including repair after demyelinating injury. Furthermore, the effects of Gas6 on STAT3 signaling and matrix MMP9 downregulation indicate potential glial cell repair and immunoregulatory roles for Gas6, indicating that Gas6-TAM signaling could be a potential therapeutic target in MS and other neuropathologies.


Reduced insulin and IGF-I signaling, not hyperglycemia, underlies the diabetes-associated depletion of interstitial cells of Cajal in the murine stomach.
Damage to interstitial cells of Cajal (ICC), pacemakers, and mediators of neuromuscular neurotransmission in the gastrointestinal tract contributes to the pathogenesis of diabetic gastroenteropathy in both patients and animal models. ICC depletion in diabetes may result from chronic hyperglycemia or lost/ineffective insulin signaling. Because independent control of insulin and glucose concentrations is difficult in chronic in vivo studies, we used long-term organotypic cultures to address this problem. Murine gastric muscles were cultured in normoglycemic or hyperglycemic basal media with or without insulin or IGF-I for 1-3 months, the time required for gastroparesis and ICC damage to develop in diabetic mice. ICC were assessed by c-Kit immunohistochemistry and quantitative analysis of c-kit expression. Electrical pacemaking was studied by intracellular recording of slow waves. ICC survived for at least 34 days in unsupplemented normoglycemic media, but their networks, c-kit expression, and slow waves were profoundly reduced after 68 days. These changes could be entirely prevented by insulin or IGF-I supplementation. ICC networks were completely resistant to hyperglycemia for at least 72 days. Thus, hyperglycemia is unlikely to be responsible for the diabetes-associated depletion of ICC. In contrast, maintenance of ICC requires insulin or IGF-I, which are reduced or ineffective in diabetes.


E2A promotes the survival of precursor and mature B lymphocytes.
The basic helix-loop-helix transcription factor E2A is an essential regulator of B lymphocyte lineage commitment and is required to activate the expression of numerous B lineage-specific genes. Studies involving ectopic expression of Id proteins, which inhibit E2A as well as other basic helix-loop-helix proteins such as HEB, suggest additional roles of E2A at later stages of B cell development. We use E2A-deficient and E2A and HEB double-deficient pre-B cell lines to directly assess the function of E2A and HEB in B cell development after lineage commitment. We show that, in contrast to the established role of E2A in lineage commitment, elimination of E2A and HEB in pre-B cell lines has only a modest negative impact on B lineage gene expression. However, E2A single and E2A and HEB double-deficient but not HEB single-deficient cell lines show dramatically enhanced apoptosis upon growth arrest. To address the possible role of E2A in the regulation of B cell survival in vivo, we crossed IFN-inducible Cre-transgenic mice to E2A conditional mice. Cre-mediated E2A deletion resulted in a block in bone marrow B cell development and a significant reduction in the proportion and total number of splenic B cells in these mice. We show that Cre-mediated deletion of E2A in adoptively transferred mature B cells results in the rapid depletion of the transferred population within 24 h of Cre induction. These results reveal that E2A is not required to maintain B cell fate but is essential in promoting pre-B and B cell survival.


Peripheral neuropathy during bortezomib treatment of multiple myeloma: a review of recent studies.
Treatment-emergent peripheral neuropathy (PN) is an important dose-limiting toxicity during treatment of multiple myeloma (MM). Bortezomib-induced PN (BIPN) occurred in 37-44% of clinical trial patients with MM, with the cumulative treatment dose as its single most significant predictor. This review discusses the clinical profile of BIPN in the treatment of MM and guidelines for its management. Lower rates of BIPN observed during treatment of solid tumors compared with rates of hematologic cancers are also discussed. Several areas of research are reviewed that may improve the management of BIPN, including co-therapies with the novel heat shock protein inhibitor tanespimycin, which appears to reduce the incidence of BIPN, and recent studies with second-generation proteasome inhibitors such as carfilzomib and NPI-0052. Adherence to the National Cancer Institute dose-modification algorithm is the most effective method for mitigating BIPN. Reversal of BIPN after treatment cessation occurs in most cases, but recovery in some patients takes as long as 1.7 years, and some individuals fail to return to baseline neurologic function. BIPN can cause a significant reduction in quality of life, primarily due to severe treatment-emergent pain. Ongoing research may provide additional information about the mechanism of BIPN and strategies to reduce PN.


Technetium Tc 99m medronate bone scanning in rhabdomyolysis.
We used technetium Tc 99m medronate (methylene diphosphonate) scanning in the treatment of seven patients with rhabdomyolysis. In three patients the intense, diffuse, symmetric nature of the scans was suggestive of an underlying metabolic muscle disorder. The scans defined the extent of the muscle injury in three patients and demonstrated muscle injury that was not initially suspected in two patients. The scan also demonstrated clinically unsuspected metastatic pulmonary and gastric calcifications in one patient. All scans showed evidence of renal dysfunction. In conclusion, these cases demonstrate that Tc 99m medronate scanning helps to define the extent and location of muscle injury in rhabdomyolysis, and that diffuse muscle uptake of Tc 99m medronate may be associated with metabolic muscle disease.


- invited review - calcium digestibility and metabolism in pigs.
Calcium (Ca) and phosphorus (P) are minerals that have important physiological functions in the body. For formulation of diets for pigs, it is necessary to consider an appropriate Ca:P ratio for an adequate absorption and utilization of both minerals. Although both minerals are important, much more research has been conducted on P digestibility than on Ca digestibility. Therefore, this review focuses on aspects that are important for the digestibility of Ca. Only values for apparent total tract digestibility (ATTD) of Ca have been reported in pigs, whereas values for both ATTD and standardized total tract digestibility (STTD) of P in feed ingredients have been reported. To be able to determine STTD values for Ca it is necessary to determine basal endogenous losses of Ca. Although most Ca is absorbed in the small intestine, there are indications that Ca may also be absorbed in the colon under some circumstances, but more research to verify the extent of Ca absorption in different parts of the intestinal tract is needed. Most P in plant ingredients is usually bound to phytate. Therefore, plant ingredients have low digestibility of P due to a lack of phytase secretion by pigs. During the last 2 decades, inclusion of microbial phytase in swine diets has improved P digestibility. However, it has been reported that a high inclusion of Ca reduces the efficacy of microbial phytase. It is possible that formation of insoluble calcium-phytate complexes, or Ca-P complexes, not only may affect the efficacy of phytase, but also the digestibility of P and Ca. Therefore, Ca, P, phytate, and phytase interactions are aspects that need to be considered in Ca digestibility studies.


Behavioural, brain and cardiac responses to hypobaric hypoxia in broiler chickens.
A novel approach to pre-slaughter stunning of chickens has been developed in which birds are rendered unconscious by progressive hypobaric hypoxia. Termed Low Atmospheric Pressure Stunning (LAPS), this approach involves application of gradual decompression lasting 280s according to a prescribed curve. We examined responses to LAPS by recording behaviour, electroencephalogram (EEG) and electrocardiogram (ECG) in individual male chickens, and interpreted these with regard to the welfare impact of the process. We also examined the effect of two temperature adjusted pressure curves on these responses. Broiler chickens were exposed to LAPS in 30 triplets (16 and 14 triplets assigned to each pressure curve). In each triplet, one bird was instrumented for recording of EEG and ECG while the behaviour of all three birds was observed. Birds showed a consistent sequence of behaviours during LAPS (ataxia, loss of posture, clonic convulsions and motionless) which were observed in all birds. Leg paddling, tonic convulsions, slow wing flapping, mandibulation, head shaking, open bill breathing, deep inhalation, jumping and vocalisation were observed in a proportion of birds. Spectral analysis of EEG responses at 2s intervals throughout LAPS revealed progressive decreases in median frequency at the same time as corresponding progressive increases in total power, followed later by decreases in total power as all birds exhibited isoelectric EEG and died. There was a very pronounced increase in total power at 50-60s into the LAPS cycle, which corresponded to dominance of the signal by high amplitude slow waves, indicating loss of consciousness. Slow wave EEG was seen early in the LAPS process, before behavioural evidence of loss of consciousness such as ataxia and loss of posture, almost certainly due to the fact that it was completely dark in the LAPS chamber. ECG recordings showed a pronounced bradycardia (starting on average 49.6s into LAPS), often associated with arrhythmia, until around 60s into LAPS when heart rate levelled off. There was a good correlation between behavioural, EEG and cardiac measures in relation to loss of consciousness which collectively provide a loss of consciousness estimate of around 60s. There were some effects of temperature adjusted pressure curves on behavioural latencies and ECG responses, but in general responses were consistent and very similar to those reported in previous research on controlled atmosphere stunning with inert gases. The results suggest that the process is humane (slaughter without avoidable fear, anxiety, pain, suffering and distress). In particular, the maintenance of slow wave EEG patterns in the early part of LAPS (while birds are still conscious) is strongly suggestive that LAPS is non-aversive, since we would expect this to be interrupted by pain or discomfort.


Oxidative stress and nutritional status in chronic obstructive pulmonary disease.
Oxidative stress and malnutrition are shown to have pathogenic effect in Chronic Obstructive Pulmonary Disease (COPD). This study was done to assess the burden of oxidative stress in COPD and to determine its relation to their nutritional status. In this cross-sectional study, 100 COPD cases from emergency and medical ward and meeting inclusion criteria, along with age, sex and occupation (mainly farmers, housewives and drivers) matched 100 controls without COPD and meeting inclusion criteria were enrolled. Oxidative stress was assessed by measuring lipid peroxidation product, Malondialdehyde (MDA) and antioxidants, like Vitamin C, E and Red Blood Cell Catalase (RBCC). Mini Nutritional Assessment (MNA) tool and Body Mass Index (BMI) were used to assess nutritional status. Chi-square test was applied for categorical variable. Student t-test was applied for comparison of means. Analysis of Variance (ANOVA) was applied for comparison between groups followed by Bonferroni post hoc analysis. Pearson correlation method was used for quantitative variables. Statistical significance was defined as p< 0.05 (two tailed). COPD cases had significantly high MDA level with low level of Vitamin E and catalase as compared to controls (p < 0.001). Most of the COPD cases were underweight (BMI ≤ 18.5 Kg/m(2)) and malnourished (MNA score less than 7). Bonferroni post-hoc analysis, showed significantly high burden of oxidative stress in underweight and malnourished cases as compared to normal weight (p < 0.05) among COPD cases. Highly significant correlation was seen between BMI and plasma MDA level (r = -0.27, p = 0.008) in COPD cases. This study shows impaired oxidant/antioxidant balance along with malnutrition and underweight in COPD, which signals for considering antioxidant therapy along with nutritional management.


Heterozygosity and instability of amplified chromosomal insertions in the radioresistant bacterium Deinococcus radiodurans.
Natural transformation, duplication insertion, and plasmid transformation in Deinococcus radiodurans, a bacterium that contains 4 to 10 chromosomes per cell, were studied. Duplication insertions were often heterozygous, with some chromosomes containing highly amplified insertions and others containing no insertions. Large amplified regions were apparently deleted by intrachromosomal recombination, generating as by-products extrachromosomal circles consisting of multiple tandem repeats of the amplified sequence. The circles were of heterogenous integer sizes, containing as many as 10 or more amplification units. Two strains that are defective in natural transformation and sensitive to DNA-damaging agents were further characterized. Both strains were defective in duplication insertion. While on strain was normal for plasmid transformation, the other was totally defective in this regard, suggesting that plasmid transfer in D. radiodurans may require recombinational functions.


Cotransfer of circular and linear prokaryotic and eukaryotic DNA sequences into mouse cells.
We have attempted to introduce some eukaryotic and prokaryotic DNA sequences into mouse fibroblasts. Purified herpes thymidine kinase gene (tk) was introduced into mouse cells. The presence of the herpes tk gene was established by gel electrophoresis, sensitivity to the purine analog acyloguanosine, and Southern blot hybridization. We utilized two different methods to introduce nonselectable markers into mouse cells. Bacterial plasmid pBR322 was ligated to herpes tk and used for transfection. All cells that were TK+ also contained the plasmid sequences. In the second method, pBR322 DNA was mixed with herpes tk DNA and presented to mouse cells. TK+ cells were tested for pBR322 sequences by blot hydridization. The frequency of unlinked cotransfer was greater than 40%. When the circular plasmid containing pBR322 and tk was used for transfection, each of the resulting transfectants acquired several copies of the plasmid. Most of the copies were associated with high molecular weight DNA in the cell. In addition, we found that some of the plasmid molecules may exist as free circular molecules. Using the nonligated cotransfer method, we introduced purified human beta-globin sequences into the recipient cells. We were unable to detect any transcripts of the human beta-globin gene at a level greater than or equal to 10 molecules per cell.


Adsorption of double-stranded ribonucleic acids (dsRNA) to iron (oxyhydr-)oxide surfaces: comparative analysis of model dsRNA molecules and deoxyribonucleic acids (DNA).
Double-stranded ribonucleic acid (dsRNA) molecules are novel plant-incorporated protectants expressed in genetically modified RNA interference (RNAi) crops. Ecological risk assessment (ERA) of RNAi crops requires a heretofore-missing detailed understanding of dsRNA adsorption in soils, a key fate process. Herein, we systematically study the adsorption of a model dsRNA molecule and of two double-stranded deoxyribonucleic acid (DNA) molecules of varying lengths to three soil iron (oxyhydr-)oxides - goethite, lepidocrocite, and hematite - over a range of solution pH (4.5-10), ionic strength (I = 10-100 mM NaCl) and composition (0.5, 1, and 3 mM MgCl2) and in the absence and presence of phosphate (0.05-5 mM) as co-adsorbate. We hypothesized comparable adsorption characteristics of dsRNA and DNA based on their structural similarities. Consistently, the three nucleic acids (NAs) showed high adsorption affinities to the iron (oxyhydr-)oxides with decreasing adsorption in the order goethite, lepidocrocite, and hematite, likely reflecting a decrease in the hydroxyl group density and positive charges of the oxide surfaces in the same order. NA adsorption also decreased with increasing solution pH, consistent with weakening of NA electrostatic attraction to and inner-sphere complex formation with the iron (oxyhydr-)oxides surfaces as pH increased. Adsorbed NA concentrations increased with increasing I and in the presence of Mg2+, consistent with adsorbed NA molecules adopting more compact conformations. Strong NA-phosphate adsorption competition demonstrates that co-adsorbates need consideration in assessing dsRNA fate in soils. Comparable adsorption characteristics of dsRNA and DNA molecules to iron (oxyhydr-)oxides imply that information on DNA adsorption to soil particle surfaces can inform dsRNA ERA.


Investigation of static structure factor in dense suspensions by use of multiply scattered light.
Near-infrared, frequency-domain photon migration measurements of phase shift are used to derive accurate values of isotropic scattering coefficients in concentrated, interacting suspensions of aqueous polystyrene microspheres with volume concentrations ranging from 1% to 45% by solids and mean diameters ranging from 135 to 500 nm. Under conditions of high ionic strength, the isotropic scattering coefficient can be quantitatively predicted by the Percus-Yevick model for hard-sphere interactions and Mie theory. In addition, the attractive interactions between scatterers arising from the addition of soluble poly(ethylene glycol) with molecular weights of 100 and 600 K cause hindered scattering. The increases in static structure and decreases in isotropic scattering coefficient agree with that predicted by Mie theory and the depletion interaction model developed by Asakura and Oosawa [J. Chem. Phys. 22, 1255 (1954)]. These results demonstrate the success of monitoring interaction between particles by use of multiple-scattered light and the necessity of incorporating models for these interactions when predicting scattering of dense, concentrated suspensions.