3D Reconstruction of the Digestive System in Octopus vulgaris Cuvier, 1797 Embryos and Paralarvae during the First Month of Life.
Octopus vulgaris aquaculture is limited due to poor biological knowledge of the paralarval stages (e.g., digestive system functionality), their nutritional requirements (e.g., adequate live diet) and standardization of rearing techniques. These factors are important in explaining the high mortality rate observed in this developmental stage under culture conditions. For a better understanding of nutrition biology of this species, we investigated the 3D microanatomy of the digestive tract of the embryo and paralarvae during the first month of life. O. vulgaris paralarvae digestive system is similar to that in the adult. The "descending branch" has a dorsal position and is formed by the buccal mass, oesophagus and crop. Ventrally, the "ascending branch" is formed by the intestine and the anus. The digestive gland, the posterior salivary glands and the inner yolk sac (in the case of the embryo and hatched paralarvae) are located between the "ascending" and "descending" branches. In the curve of the U-shaped digestive tract, a caecum and the stomach can be found. The reconstructions reveal that anatomically the digestive system is already complete when the paralarvae hatch. The reconstruction of the buccal mass at different post-hatching days has demonstrated that all the necessary structures for food intake are present. However, the radula surface in contact with the pharynx is very small on the first day of life. Although the digestive system has all the structures to feed, the digestive gland and radula take longer to reach full functionality. We have established four development periods: embryonic, early post-hatching, late post-hatching and juvenile-adult. The differentiation between these periods was done by type of feeding (endogenous or exogenous), the state of maturation and hence functionality of the digestive gland, type of growth (linear, no net, or exponential), and measurement of the arm lengths with respect to the mantle length. 3D reconstruction represents a new tool to study the morphology and functionality of the cephalopod digestive system during the first days of life.


Sculpting the valley in the radius distribution of small exoplanets as a by-product of planet formation: the core-powered mass-loss mechanism.
Recent observations revealed a bimodal radius distribution of small, short-period exoplanets with a paucity in their occurrence, a radius 'valley', around 1.5-2.0 R⊕. In this work, we investigate the effect of a planet's own cooling luminosity on its thermal evolution and atmospheric mass loss (core-powered mass-loss) and determine its observational consequences for the radius distribution of small, close-in exoplanets. Using simple analytical descriptions and numerical simulations, we demonstrate that planetary evolution based on the core-powered mass-loss mechanism alone (i.e. without any photoevaporation) can produce the observed valley in the radius distribution. Our results match the valley's location, shape and slope in planet radius-orbital period parameter space, and the relative magnitudes of the planet occurrence rate above and below the valley. We find that the slope of the valley is, to first order, dictated by the atmospheric mass-loss time-scale at the Bondi radius and given by d logR p/d logP ≃ 1/(3(1 - β)) that evaluates to -0.11 for β ≃ 4, where M c/M⊕ = (R c/R ⊕) β (ρ c*/ρ ⊕) β/3 is the mass-radius relation of the core. This choice for β yields good agreement with observations and attests to the significance of internal compression for massive planetary cores. We further find that the location of the valley scales as ρ c ∗ - 4 ∕ 9 and that the observed planet population must have predominantly rocky cores with typical water-ice fractions of less than ~20 per cent. Furthermore, we show that the relative magnitude of the planet occurrence rate above and below the valley is sensitive to the details of the planet-mass distribution but that the location of the valley is not.


Optimization of the piggyBac transposon using mRNA and insulators: toward a more reliable gene delivery system.
Integrating and expressing stably a transgene into the cellular genome remain major challenges for gene-based therapies and for bioproduction purposes. While transposon vectors mediate efficient transgene integration, expression may be limited by epigenetic silencing, and persistent transposase expression may mediate multiple transposition cycles. Here, we evaluated the delivery of the piggyBac transposase messenger RNA combined with genetically insulated transposons to isolate the transgene from neighboring regulatory elements and stabilize expression. A comparison of piggyBac transposase expression from messenger RNA and DNA vectors was carried out in terms of expression levels, transposition efficiency, transgene expression and genotoxic effects, in order to calibrate and secure the transposition-based delivery system. Messenger RNA reduced the persistence of the transposase to a narrow window, thus decreasing side effects such as superfluous genomic DNA cleavage. Both the CTF/NF1 and the D4Z4 insulators were found to mediate more efficient expression from a few transposition events. We conclude that the use of engineered piggyBac transposase mRNA and insulated transposons offer promising ways of improving the quality of the integration process and sustaining the expression of transposon vectors.


Enhanced radiation and chemotherapy-mediated cell killing of human cancer cells by small inhibitory RNA silencing of DNA repair factors.
Recent developments in the use of small inhibitory RNA molecules (siRNAs) to inhibit specific protein expression have highlighted the potential use of siRNA as a therapeutic agent. The double-strand break signaling/repair proteins ATM, ATR, and DNA-dependent protein kinase catalytic subunit (DNA-PK(cs)) are attractive targets to confer enhanced radio and chemosensitivity to tumor cells. We have designed and exogenously delivered plasmids encoding siRNAs targeting these critical kinases to human cancer cells to assess the feasibility of this concept as a clinically translatable experimental therapeutic. siRNA led to a approximately 90% reduction in target protein expression. siRNAs targeting ATM and DNA-PK(cs) gave rise to a dose-reduction factor of approximately 1.4 compared with untransfected and control vector-transfected cells at the clinically relevant radiation doses. This was greater than the radiosensitivity achieved using the phosphatidylinositol 3'-kinase inhibitor Wortmannin or DNA-PK(cs) competitive inhibitor LY294002. A similar increased sensitivity to the alkylating agent methyl methanesulfonate (MMS) was also observed for siRNA-mediated ATR silencing. Together, these data provide strong evidence for the potential use of siRNA as a novel radiation/chemotherapy-sensitizing agent.


Brain plasticity and intellectual ability are influenced by shared genes.
Although the adult brain is considered to be fully developed and stable until senescence when its size steadily decreases, such stability seems at odds with continued human (intellectual) development throughout life. Moreover, although variation in human brain size is highly heritable, we do not know the extent to which genes contribute to individual differences in brain plasticity. In this longitudinal magnetic resonance imaging study in twins, we report considerable thinning of the frontal cortex and thickening of the medial temporal cortex with increasing age and find this change to be heritable and partly related to cognitive ability. Specifically, adults with higher intelligence show attenuated cortical thinning and more pronounced cortical thickening over time than do subjects with average or below average IQ. Genes influencing variability in both intelligence and brain plasticity partly drive these associations. Thus, not only does the brain continue to change well into adulthood, these changes are functionally relevant because they are related to intelligence.


A comparison of hip fracture incidence rates among elderly in Sweden by latitude and sunlight exposure.
Research has shown that hip fracture risk increases with latitude; hypothetically due to reduced sunlight exposure and its effect on bone quality. Sweden, with large differences in latitude and UV radiation, is ideal to study in order to analyse the association between latitude and UV radiation on age- and sex-specific hip fracture rates among elderly. Aggregated (2006-2008) age- and sex-specific hip fracture data was obtained for each Swedish municipality as well as the municipality's latitudinal coordinates and aggregated (2006-2008) UV radiation levels. Pearson correlations were calculated between hip fracture incidence rates, latitude and UV radiation. Independent t tests were calculated on tertile-categorized latitudinal data in order to investigate the difference in hip fracture risk between these categories. Statistically significant correlations were seen in all groups between hip fracture incidence rates and latitude as well as UV radiation. The independent t tests showed that this correlation was mainly due to high incidence rates in high latitude municipalities. Statistically significant correlations are seen between hip fracture incidence rates and latitude as well as UV radiation in Sweden and the northern parts of Sweden have an increased risk of hip fractures compared to the middle and southern parts. To our knowledge this is the first study using a national discharge register that shows this relationship and provides a starting point for further research to investigate why populations in northern Sweden have a higher risk of hip fractures compared to other Swedish regions.


A comprehensive description of functioning and disability in children with velopharyngeal insufficiency.
Children with velopharyngeal insufficiency (VPI) experience functional impairments in a variety of areas that extend beyond the primary physical impairment associated with this disorder. At present, the physical deficits associated with VPI have been studied extensively; however, a comprehensive description of social and communicative participation in this population is needed. Therefore, a biopsychosocial framework such as the International Classification of Functioning, Disability and Health for Children and Youth (ICF-CY), may offer an enhanced understanding of the daily experiences of children with VPI. Specifically, the ICF-CY framework is intended to model complex nonlinear systems, and as such, to describe functioning as the interaction of multiple components from which a limitation in communicative participation may emerge. This paper describes how the ICF-CY framework can be utilized to comprehensively describe functioning and disability in children with VPI by describing the interaction of components of this framework. As a result of this activity, the reader will be able to: (1) discuss the utility of the ICF-CY in describing the multi-dimensional nature of velopharyngeal insufficiency (VPI); (2) describe interrelationships between functioning and disability in children with VPI; and (3) identify how limitations in communicative participation may emerge from the interaction of components of the ICF-CY in children with VPI.


[Complicated Meckel's diverticulum].
In this study we present our experience in the surgical treatment of complicated Meckel's diverticulum. The data of eight patients who underwent surgery due to complications of Meckel's diverticulum between 1994-2001 was retrospectively assessed. There were six males and two females with a mean age of 31 years (range 13 to 65). Preoperative diagnoses were acute surgical abdomen in six and incarcerated inguinal hernias in two patients. Intraoperative diagnoses were as follows; Littre's hernia in two, diverticulitis in two, perforation of the diverticulum in one and intestinal obstruction in three patients (there was a band extending from diverticulum to the umbilicus in two patients and a mesodiverticular band in the remaining one). While diverticulectomies were performed in five patients, three had small bowel resections. The mean diameter of the diverticula was 2.3 cm (range 2 to 4) and the mean length was 3.5 cm (range 3 to 8). Postoperative intestinal obstruction was observed in one patient who had underwent diverticulectomy and subsequently a small bowel resection was performed. Being aware of the complications of the Meckel's diverticulum is necessary in correct timing of the surgery and selecting the proper incision in patients with acute abdomen. This will result in decreased morbidity.


A national survey of surgical activity in hospitals in Malawi.
Malawi is a poor country with few doctors. It has 21 district hospitals, all of which have operating theatres but none of which has a surgeon. Most district hospitals are manned by a single doctor and two or more paramedic clinical officers. In 2003, a total of 28,594 operations were performed in district hospitals. In all, 12,506 (44%) of these were obstetric or gynaecological procedures, but only 821 (3%) were general surgical cases. It appears that district clinicians are happy to attempt emergency obstetric and gynaecological cases but tend to refer emergency general surgical cases.


The choice of contrast agents in the cardiac catheterization laboratory.
The availability of newer and more expensive low osmolar contrast agents for cardiac angiography has forced a comparison with the standard ionic contrast agents. The milder hemodynamic effects of low osmolar agents make them the contrast of choice for the sickest patients; however, the existence of a reduction in the rate of anaphylactoid reactions or death during catheterization seems more doubtful. Guidelines for the selective use of the agents in the cardiac catheterization laboratory are offered, based on one hospital's experience of more than 20,000 procedures.


Synthesis and application of a new antibacterial surfactant from apricot kernel oil.
Food emulsifier are mostly prepared from a lipophilic lipid tail with a hydrophilic sugar head. In this study, the lipophilic tail was obtained from apricot kernels, which are food waste, and the hydrophilic head was gluconic acid instead of sugar, in order to draw attention to the non-cyclic poly hydroxyl compounds. Thus, oleic acid of apricot kernel was used as the lipophilic moiety of the prepared surfactant. So, apricot kernel was grinned and dried, oil was extracted using soxhlet apparatus, Physical and chemical parameters and fatty acids composition of the extracted oil had been determined. The extracted oil was then hydrolyzed into glycerol and a mixture of free fatty acids. The fatty acids mixture was separated. Then, oleic acid was extracted individually in pure form using supercritical CO2 extractor, it was then confirmed according to its melting point, Gas chromatography-mass spectrometry (GC-MS) after esterification, elemental analysis, Proton nuclear magnetic resonance (H1NMR), and mass spectrometry (MS) to detect the corresponding molecular ion peak. The pure individual oleic acid was converted to hydroxy stearic acid, which was then converted to an amphiphilic compound (surfactant) via esterification reaction with the hydrophilic gluconic acid, and afforded a new surfactant known as 2,3,4,5-tetrahydroxy-6-((9-((-2,3,4,5,6-pentahydroxyhexanoyl) oxy)octadecanoyl) oxy)hexanoic acid or stearyl gluconate for simplification. The structures elucidation of all synthesized compound was established according to elemental analysis and spectral data (Fourier transform infrared IR, 1H NMR, 13C NMR and MS). Moreover, the prepared compound was tasted for its antibacterial activity, and showed good activities against some types of bacteria. The surface-active properties, foamability, foaming stability and emulsion stability of stearyl gluconate were studied and compared with the properties of the well-known surfactant sucrose stearate, and it was clear that, the activity of stearyl gluconate as a surfactant was higher than that of sucrose stearate. Moreover, establishment of safety of this compound was performed using albino rats by acute oral toxicity and kidney and liver functions of these mice. On the other hand, the prepared surfactant was used in the production of low fat-free cholesterol mayonnaise as egg replacer. Texture properties and the sensory evaluation of the prepared mayonnaise showed that the properties were improved by using the new prepared surfactant. Thus, the prepared gluconyl stearate can be used as a safe food additive.


Human alpha-galactosidase A: glycosylation site 3 is essential for enzyme solubility.
Human alpha-galactosidase A (EC 188.8.131.52; alpha-Gal A) is the homodimeric glycoprotein that hydrolyses the terminal alpha-galactosyl moieties from glycolipids and glycoproteins. The type, site occupancy and function of the N-linked oligosaccharide chains on this lysosomal hydrolase were determined. Endoglycosidase treatment of the purified recombinant enzyme and mutagenesis studies indicated that three (Asn-139, Asn-192 and Asn-215) of the four potential N-glycosylation consensus sequences were occupied by complex, high-mannose and hybrid-type oligosaccharides respectively. When expressed in COS-1 cells, glycoforms with glycosylation site 1 or 2 obliterated had more than 70% of wild-type activity, and both glycoforms were secreted. In contrast, the glycoform with only site 3 eliminated had decreased activity (less than 40%); little, if any, was secreted. Expressed mutant glycoforms in which site 3 and site 1 or 2 were obliterated had little, if any, intracellular or secreted enzymic activity, and immunofluorescence microscopy revealed that the expressed mutant glycoforms were retained in the endoplasmic reticulum, presumably where they were degraded. Thus glycosylation at site 3 was crucial to the formation of soluble, active enzyme, as well as transport to the lysosome. Absence of the site 3 hybrid-type oligosaccharide exposed an adjacent, normally protected, hydrophobic region, resulting in aggregation of the enzyme polypeptide in the endoplasmic reticulum. In support of this concept, endoglycosidase H-treated enzyme or mannose-terminated enzyme expressed in Autographa californica cells also aggregated when concentrated, emphasizing that site 3 occupancy by a hybrid-type oligosaccharide was required for enzyme solubility.


Effect of litter size and bacitracin administration on tissue protein synthesis of lactating rabbit does.
Bacitracin is an antibiotic used in rabbit husbandry to control microbial digestive pathologies. Collateral effects on absorption and mucosal development have been reported and these may impact on protein metabolism. This study aims to analyse the effect of the antibiotic on protein synthesis in lactating does because mammary gland metabolism and milk output should provide a sensitive index of any undesirable action of bacitracin. Rates of protein synthesis were measured in mammary gland, liver, intestinal mucosa and muscle of lactating rabbits does by injecting a flooding dose of [(2)H(5)]phenylalanine into the auricular artery of two groups (each n = 8) of New Zealand White does fed different experimental diets. The control group (C) received the basal diet and the bacitracin group (B) ingested the same diet but supplemented with bacitracin (100 mg/kg). Animals received the experimental diet from day 28 of pregnancy until day 26 of lactation when they were slaughtered. Just after birth, litter size was adjusted by cross-fostering either to five or nine pups (four does per dietary treatment). The relative weight of the liver tended to be greater in those females receiving the B diet (27 v. 22.5 g/kg BW; P < 0.07), while diet did not affect mammary gland weight (255.7 ± 10.59 g). Fractional protein synthesis rate (FSR) was higher for intestinal mucosa (duodenum; 51.7% ± 2.09%/day) followed by mammary gland and liver (38.29 ± 2.62%/day and 40.2 ± 1.98%/day, respectively), and the lowest value was observed in muscle (2.92 ± 0.26%/day; P < 0.0001). Bacitracin treatment lowered FSR in the mammary gland by 23% (P = 0.024) and this was independent of litter size. Conversely, FSR in the duodenum was not affected by antibiotic treatment but reduced by 15% (P = 0.021) for the larger litter size.


Natural and synthetic flavonoid derivatives as new potential tyrosinase inhibitors: a systematic review.
Tyrosinase is a multifunctional glycosylated and copper-containing oxidase that is highly prevalent in plants and animals and plays a pivotal role in catalyzing the two key steps of melanogenesis: tyrosine's hydroxylation to dihydroxyphenylalanine (DOPA), and oxidation of the latter species to dopaquinone. Melanin guards against the destructive effects of ultraviolet radiation which is known to produce considerable pathological disorders such as skin cancer, among others. Moreover, the overproduction of melanin can create aesthetic problems along with serious disorders linked to hyperpigmented spots or patches on skin. Several skin-whitening products which reduce melanogenesis activity and alleviate hyperpigmentation are commercially available. A few of them, particularly those obtained from natural sources and that incorporate a phenolic scaffold, have been exploited in the cosmetic industry. In this context, synthetic tyrosinase inhibitors (TIs) with elevated efficacy and fewer side effects are direly needed in the pharmaceutical and cosmetic industries owing to their protective effect against pigmentation and dermatological disorders. Furthermore, the biological significance of the chromone skeleton and its associated medicinal and bioactive properties has drawn immense interest and inspired many researchers to design and develop novel anti-tyrosinase agents based on the flavonoid core (2-arylchromone). This review article is oriented to provide an insight and a deeper understanding of the tyrosinase inhibitory activity of an array of natural and bioinspired phenolic compounds with special emphasis on flavonoids to demonstrate how the position of ring substituents and their interaction with tyrosinase could be correlated with their effectiveness or lack thereof against inhibiting the enzyme.


Knowledge and attitudes about transcranial magnetic stimulation among psychiatrists in China.
Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive form of brain stimulation. It has been used in many mental health institutions to treat mental disorders worldwide. However, comprehensive knowledge about rTMS is not yet widespread among psychiatrists. The present study assessed psychiatrists' knowledge and attitudes about rTMS in China and investigated related factors. A quantitative observational cross-sectional study was conducted using an online survey. The sample consisted of 522 psychiatrists. Multinomial logistic regression and multiple linear regression analyses were used to explore factors that contributed to psychiatrists' knowledge about rTMS. We also ascertained psychiatrists' attitudes about rTMS and provide recommendations for the more widespread use of rTMS. The majority of respondents (86.4%) reported having access to rTMS at their institution. A total of 379 psychiatrists (72.6%) knew that rTMS was approved by the United States Food and Drug Administration for treatment-resistant depression. Univariate logistic regression indicated that psychiatrists who were older, had a senior professional title, worked more years, had an onsite clinical rTMS program in their hospital, and received formal training in theory and application (all p < 0.05) were more likely to know that rTMS was approved by the Food and Drug Administration for the treatment of depression. The percentages of respondents who knew most or all indications, the mechanism of action, parameter settings, adverse reactions were 51.9, 40.2, 27.4, and 41.4%. Linear regression showed that formal training in rTMS theory and practice were associated with higher knowledge scores (all p < 0.05). Most of the subjects had negative attitudes about using rTMS to treat mental disorders. When asked about their attitudes about continuing rTMS education, nearly all of the respondents indicated that they were willing to pursue continuing training in rTMS in the future. Many psychiatrists had an insufficient level of knowledge about rTMS and negative attitudes about rTMS. Psychiatrists who had formal rTMS training experience had higher levels of rTMS knowledge. rTMS training and relevant policy making should be strengthened.


Muscle strength, volume and activation following 12-month resistance training in 70-year-old males.
In elderly males muscle plantar flexor maximal voluntary contraction (MVC) torque normalised to muscle volume (MVC/VOL) is reduced compared to young males as a result of incomplete muscle activation in the elderly. The aim of the present study was to determine the influence of a 12-month resistance training programme on muscle volume, strength, MVC/VOL, agonist activation and antagonist coactivation of the plantarfexors in elderly males. Thirteen elderly males aged 70 years and over (range 70-82 years), completed a 12-month whole body resistance-training programme (TRN), training three times a week. Another eight males (range 18-30 years), who maintained their habitual physical activity for the same 12-month period as the TRN group acted as controls (CTRL). Isometric plantarflexor maximal voluntary contraction (MVC) torque increased in the TRN group by 20% (P < 0.01), from 113.1 +/- 22.0 Nm to 141.5 +/- 19.2 Nm. Triceps surae volume (TS VOL) assessed using MRI, increased by 12%, from 796.3 +/- 78.9 cm(3) to 916.8 +/- 144.4 cm(3) . PF activation, measured using supramaximal double twitch interpolation, increased from 83.6+/-11.0% pre training, to 92.1 +/- 7.6% post training (P < 0.05). Dorsiflexion MVC and antagonist coactivation (assessed using surface electromyography) did not change with training. Plantarflexor MVC torque normalized for triceps surae muscle volume (MVC/VOL) was 142.6 +/- 32.4 kN m(-2) before training and 157.0 +/- 27.9 kN m(-2) after training (a non-significant increase of 8%). No significant change in any measurement was observed in the CTRL group. This study has shown that the gain in muscle strength in response to long-term (12-month) training in older men is mostly accounted for by an increased muscle volume and activation.


Study of immunoelectron microscopic localization of cathepsin K in osteoclasts and other bone cells in the mouse femur.
The localization of cathepsin K protein in mouse osteoclasts was examined by immunolight and immunoelectron microscopy using the avidin-biotin-peroxidase complex method with anti-cathepsin K (mouse) antibody. With light microscopy, a strong immunoreaction for cathepsin K was found extracellularly along the bone and cartilage resorption lacunae and detected intracellularly in vesicles, granules, and vacuoles throughout the cytoplasm of multinuclear osteoclasts and chondroclasts attached to the surface of the bone or cartilage. Mononuclear cells, probably preosteoclasts, some distance from the bone also contained a few cathepsin K-positive vesicles and granules. Cathepsin K was sometimes found in the cisternal spaces of the rough endoplasmic reticulum and vesicles of the Golgi apparatus with electron microscopy of the basolateral region of the osteoclasts. Cathepsin K-positive vesicles and granules as lysosomal compartments were present in various stages of fusion with vacuoles as endosomal compartments that contained fragmented cathepsin K-negative fibril-like structures. Some of the vacuoles (endolysosomes), which seemed to be formed by this process of fusion, contained cathepsin K-positive vesicles and fibril-like structures that did not show the regular cross striation of type I collagen fibrils. In the apical region of the osteoclasts, cathepsin K-positive vesicles and pits had already fused with or were in the process of fusing with the ampullar extracellular spaces. There were large deposits of cathepsin K on fragmented fibril-like structures without regular cross striation in the extracellular spaces, as well as on and between the cytoplasmic processes of the ruffled border. There were also extensive deposits of cathepsin K on the type I collagen fibrils with cross striation in the bone resorption lacunae. Osteoblasts and osteocytes were negative for cathepsin K. In the immunocytochemical controls, no immunoreaction was found in the osteoclasts or preosteoclasts, or on the collagen fibrils in the resorption lacunae. The results indicate that cathepsin K is produced in mature osteoclasts attached to the bone and secreted into the bone resorption lacunae. The findings suggest that cathepsin K participates in the extracellular degradation of collagen fibrils in the resorption lacunae and in the subsequent degradation of the fragmented fibrils in the endolysosomes. It is also suggested that cathepsin K degrades the organic cartilage matrix.


Microglia protect neurons against ischemia by synthesis of tumor necrosis factor.
Microglia and infiltrating leukocytes are considered major producers of tumor necrosis factor (TNF), which is a crucial player in cerebral ischemia and brain inflammation. We have identified a neuroprotective role for microglial-derived TNF in cerebral ischemia in mice. We show that cortical infarction and behavioral deficit are significantly exacerbated in TNF-knock-out (KO) mice compared with wild-type mice. By using in situ hybridization, immunohistochemistry, and green fluorescent protein bone marrow (BM)-chimeric mice, TNF was shown to be produced by microglia and infiltrating leukocytes. Additional analysis demonstrating that BM-chimeric TNF-KO mice grafted with wild-type BM cells developed larger infarcts than BM-chimeric wild-type mice grafted with TNF-KO BM cells provided evidence that the neuroprotective effect of TNF was attributable to microglial- not leukocyte-derived TNF. In addition, observation of increased infarction in TNF-p55 receptor (TNF-p55R)-KO mice compared with TNF-p75R and wild-type mice suggested that microglial-derived TNF exerts neuroprotective effects through TNF-p55R. We finally report that TNF deficiency is associated with reduced microglial population size and Toll-like receptor 2 expression in unmanipulated brain, which might also influence the neuronal response to injury. Our results identify microglia and microglial-derived TNF as playing a key role in determining the survival of endangered neurons in cerebral ischemia.


Advantages of total parathyroidectomy in patients with secondary hyperparathyroidism induced by end stage renal disease.
Secondary hyperparathyroidism, as a result of chronic kidney disease could be treated medically or surgically. When pharmacotherapy fails, patients undergo surgery - parathyroidectomy, the curative treatment of secondary hyperparathyroidism (SHPT). There are currently 3 accepted surgical techniques, each with supporters or opponents - total parathyroidectomy, subtotal parathyroidectomy and parathyroidectomy with immediate autotransplantation. In this paper we described our experience on a series of 160 consecutive patients diagnosed with secondary hyperparathyroidism who underwent surgery, in 27 cases it was totalization of the intervention (patients with previously performed subtotal parathyroidectomy or with supernumerary glands and SHPT recurrence). We routinely perform total parathyroidectomy, the method that we believe offers the best results. The group of patients was studied according to demographic criteria, paraclinical balance, clinical symptomatology, pre- and postoperative iPTH (intact parathormone) values, SHPT recurrence, number of reinterventions. In 31 cases we found gland ectopy and in 15 cases we discovered supernumerary parathyroids. A percentage of 96.24% of patients with total parathyroidectomy did not show recurrence. After analyzing the obtained results, our conclusion was that total parathyroidectomy is the intervention of choice for patients suffering from secondary hyperparathyroidism when pharmacotherapy fails in order to prevent recurrence of the disease and to correct the metabolic parameters.


Forming design teams to develop healthcare information systems.
Healthcare information systems are assuming an increasingly critical role in providing quality patient care in an effective and efficient manner. However, the success of these systems in achieving these goals remains a lingering concern. Consequently, investigating and devising strategies to enhance the likelihood of success of a healthcare information system continues to draw research interest. One strategy recommended by both researchers and practitioners alike is the participation of the target users in the design and development of the information system. However, practical considerations mandate representative, rather than universal, participation of users. Unfortunately, the information systems literature offers few guidelines for selecting user representatives to serve on a design team. This lack of guidelines easily results in system designers talking with the wrong users or managers assigning the wrong users to the design team. On the basis of the theoretical paradigms underlying the user participation and design team concepts, the authors examined and derived user characteristics that are considered the most critical criteria for selecting user members of a design team. They then report on a field survey they conducted to validate the derived criteria in healthcare information systems context. The authors conclude that the system-related functional expertise should be the primary criterion employed to select healthcare personnel to participate in system design and development. Other criteria, such as users' communication skills, computing backgrounds, and personality traits, should be given secondary considerations. Ignoring these guidelines can render user participation superfluous, resulting in system failures.


Management of metastatic kidney cancer in the era of personalized medicine.
Patients with localized renal cell cancer (RCC) are often cured following surgical resection. However, a significant proportion of patients will experience recurrence or present with metastatic disease at distant sites and may be deemed incurable. The worldwide incidence of RCC is rising, affecting more than 271,000 people and resulting in 116,000 deaths each year. Unfortunately, advanced RCC is typically resistant to classical chemotherapy and radiotherapy. Previously, non-specific immunotherapies such as interleukin-2 and interferon were used in hopes of improving cancer immunity, leading to rare but durable responses. However, enthusiasm for these immunotherapies has waned due to limited patient responses, their excessive toxicities, and the emergence of alternative targeted therapies that have resulted in improved clinical endpoints for patients with metastatic RCC (mRCC). Strides in targeted treatment can be attributed to an improved understanding of the molecular underpinnings that cause and drive the progression of renal cell cancers. More recently, interest in immunotherapies has resurfaced, as agents inhibiting specific checkpoints involved in cancer immune evasion have demonstrated promising activity in patients with mRCC. Here we review the novel targeted agents, biomarkers and immunotherapies that promise to change the clinical outcomes for patients with advanced RCC.


Regional distribution of cortical interneurons and development of inhibitory tone are regulated by Cxcl12/Cxcr4 signaling.
Interneurons are born in subcortical germinative zones and tangentially migrate in multiple streams above and below the developing cortex, and then, at the appropriate developmental stage, migrate radially into the cortex. The factors that control the formation of and the timing of exit from the streams remain obscure; moreover, the rationale for this complicated developmental plan is unclear. We show that a chemokine, Cxcl12, is an attractant for interneurons during the stage of stream formation and tangential migration. Furthermore, the timing of exit from the migratory streams accompanies loss of responsiveness to Cxcl12 as an attractant. Mice with mutations in Cxcr4 have disorganized migratory streams and deletion of Cxcr4 after the streams have formed precipitates premature entry into the cortical plate. In addition, constitutive deletion of Cxcr4 specifically in interneurons alters the regional distribution of interneurons within the cortex and leads to interneuron laminar positioning defects in the postnatal cortex. To examine the role of interneuron distribution on the development of cortical circuitry, we generated mice with focal defects in interneuron distribution and studied the density of postnatal inhibitory innervation in areas with too many and too few interneurons. Interestingly, alterations in IPSC frequency and amplitude in areas with excess interneurons tend toward normalization of inhibitory tone, but in areas with reduced interneuron density this system fails. Thus, the processes controlling interneuron sorting, migration, regional distribution, and laminar positioning can have significant consequences for the development of cortical circuitry and may have important implications for a range of neurodevelopmental disorders.


Conversion and degradation pathways of sulfoximines.
In recent years, sulfoximines have received significant attention in both academia and industry. This development has been reinforced by the introduction of new methods for preparing and efficiently accessing key sulfur-based reagents allowing further functionalisations. For all fields and in particular those related to pharmaceutical and agrochemical applications, the knowledge of potential degradation pathways of sulfoximines is vital. Being aware of possible metabolites minimises the risks of late stage failures. In this review, we describe chemical transformations in the laboratory leading to sulfoximine degradations. Those include reductive and oxidative pathways as well as C-S bond cleavage reactions. In a wide frame the intention of this summary is to provide a better understanding of decomposition-like reactions of sulfoximines thereby enabling a more focussed search and analysis of byproducts and potential metabolites in biorelevant systems.


Molecular characterization of canine BCR-ABL-positive chronic myelomonocytic leukemia before and after chemotherapy.
Genetic aberrations linked to tumorigenesis have been identified in both canine and human hematopoietic malignancies. While the response of human patients to cancer treatments is often evaluated using cytogenetic techniques, this approach has not been used for dogs with comparable neoplasias. The aim of this study was to demonstrate the applicability of cytogenetic techniques to evaluate the cytogenetic response of canine leukemia to chemotherapy. Cytology and flow cytometric techniques were used to diagnose chronic myelomonocytic leukemia in a dog. High-resolution oligonucleotide array comparative genomic hybridization (oaCGH) and multicolor fluorescence in situ hybridization (FISH) were performed to identify and characterize DNA copy number aberrations (CNAs) and targeted structural chromosome aberrations in peripheral blood WBC at the time of diagnosis and following one week of chemotherapy. At the time of diagnosis, oaCGH indicated the presence of 22 distinct CNAs, of which trisomy of dog chromosome 7 (CFA 7) was the most evident. FISH analysis revealed that this CNA was present in 42% of leukemic cells; in addition, a breakpoint cluster region-Abelson murine leukemia viral oncogene homolog (BCR-ABL) translocation was evident in 17.3% of cells. After one week of treatment, the percentage of cells affected by trisomy of CFA7 and BCR-ABL translocation was reduced to 2% and 3.3%, respectively. Chromosome aberrations in canine leukemic cells may be monitored by molecular cytogenetic techniques to demonstrate cytogenetic remission following treatment. Further understanding of the genetic aberrations involved in canine leukemia may be crucial to improve treatment protocols.


Spatially regularized mixture model for lesion segmentation with application to stroke patients.
In medical imaging, lesion segmentation (differentiation between lesioned and non-lesioned tissue) is a crucial and difficult task. Automated segmentation algorithms based on intensity analysis have been already proposed and recent developments have shown that integrating spatial information enhances automatic image segmentation. However, spatial modeling is often limited to short-range spatial interactions that deal only with noise or small artifacts. Previous tissue alterations (e.g. white matter disease (WMD)) similar in intensity with the lesion of interest require a broader-scale approach to be corrected. On the other hand, imaging techniques offer now a multiparametric voxel characterization that may help differentiating lesioned from non-lesioned voxels. We developed an unsupervised multivariate segmentation algorithm based on finite mixture modeling that incorporates spatial information. We extended the usual spatial Potts model to the regional scale using a 'multi-order' neighborhood potential, with internal adjustment of the regional scale according to the lesion size. We validate the ability of this new algorithm to deal with noise and artifacts (linear and spherical) using artificial data. We then assess its performance on real magnetic resonance imaging brain scans of stroke patients with history of WMD and show that regional regularization was able to remove large-scale WMD artifacts.


Development of transient outward currents coupled with Ca2+-induced Ca2+ release mediates oscillatory membrane potential in ascidian muscle cells.
Isolated ascidian Halocynthia roretzi blastomeres of the muscle lineage exhibit muscle cell-like excitability on differentiation despite the arrest of cell cleavage early in development. This characteristic provides a unique opportunity to track changes in ion channel expression during muscle cell differentiation. Here, we show that the intrinsic membrane property of ascidian cleavage-arrested muscle-type cells becomes oscillatory by expressing transient outward currents (I(to)) activated by Ca(2+)-induced Ca(2+) release (CICR) in a maturation-dependent manner. In current-clamp mode, most day 4 (72 h after fertilization) cleavage-arrested muscle cells exhibited an oscillatory membrane potential of -20 mV at 15 Hz, whereas most day 3 (48 h after fertilization) cells exhibited a spiking pattern. In voltage-clamp mode, the day 4 cells exhibited prominent transient outward currents that were not present in day 3 cells. I(to) was abolished by the application of 10 mM caffeine, implying that CICR was involved in I(to) activation. I(to) was based on K(+) efflux and sensitive to tetraethylammonium and some Ca(2+)-activated K(+) channel inhibitors. We found a 60-pS single channel conductance that was activated by local Ca(2+) release in ascidian muscle cell. Voltage-clamp recording with an oscillatory waveform as a command pulse showed that CICR-activated K(+) currents were activated during the falling phase of the membrane potential oscillation. These results suggest that developmental expression of CICR-activated K(+) current plays a role in the maturation of larval locomotion by modifying the intrinsic membrane excitability of muscle cells.


Allergic sensitization and microbial load--a comparison between Finland and Russian Karelia.
Epidemiological data have indicated that some infections are associated with a low risk of allergic diseases, thus supporting the idea (hygiene hypothesis) that the microbial load is an important environmental factor conferring protection against the development of allergies. We set out to test the hygiene hypothesis in a unique epidemiological setting in two socio-economically and culturally markedly different, although genetically related, populations living in geographically adjacent areas. The study cohorts included 266 schoolchildren from the Karelian Republic in Russia and 266 schoolchildren from Finland. The levels of total IgE and allergen-specific IgE for birch, cat and egg albumen were measured. Microbial antibodies were analysed against enteroviruses (coxsackievirus B4), hepatitis A virus, Helicobacter pylori and Toxoplasma gondii. Although total IgE level was higher in Russian Karelian children compared to their Finnish peers, the prevalence of allergen-specific IgE was lower among Russian Karelian children. The prevalence of microbial antibodies was, in turn, significantly more frequent in the Karelian children, reflecting the conspicuous difference in socio-economic background factors. Microbial infections were associated with lower risk of allergic sensitization in Russian Karelian children, enterovirus showing the strongest protective effect in a multivariate model. The present findings support the idea that exposure to certain infections, particularly in childhood, may protect from the development of atopy. Enterovirus infections represent a new candidate to the list of markers of such a protective environment. However, possible causal relationship needs to be confirmed in further studies.


Bio-Morphological Reaction of Human Periodontal Ligament Fibroblasts to Different Types of Dentinal Derivates: In Vitro Study.
Understanding the biological and morphological reactions of human cells towards different dentinal derivate grafting materials is fundamental for choosing the type of dentin for specific clinical situations. This study aimed to evaluate human periodontal ligament fibroblasts (hPLF) cells exposed to different dentinal derivates particles. The study design included the in vitro evaluation of mineralized dentine (SG), deproteinized and demineralized dentine (DDP), and demineralized dentine (TT) as test materials and of deproteinized bovine bone (BIOS) as the positive control material. The materials were kept with the hPLF cell line, and the evaluations were made after 24 h, 72 h, and 7 days of in vitro culture. The evaluated outcomes were proliferation by using XTT assays, the morphological characteristics by light microscopy (LM) and by the use of scanning electron microscopy (SEM), and adhesion by using confocal microscopy (CLSM). Overall, the experimental materials induced a positive response of the hPLFs in terms of proliferation and adhesion. The XTT assay showed the TT, and the SG induced significant growth compared to the negative control at 7 days follow-up. The morphological data supported the XTT assay: the LM observations showed the presence of densely packed cells with a modified shape; the SEM observations allowed the assessment of how fibroblasts exposed to DDP and TT presented cytoplasmatic extensions; and SG and BIOS also presented the thickening of the cellular membrane. The CLMS observations showed the expression of the proliferative marker, as well as and the expression of cytoskeletal elements involved in the adhesion process. In particular, the vinculin and integrin signals were stronger at 72 h, while the actin signal remained constantly expressed in all the follow-up of the sample exposed to SG material. The integrin signal was stronger at 72 h, and the vinculin and actin signals were stronger at 7 days follow-up in the sample exposed to DDP material. The vinculin and integrin signals were stronger at 72 h follow-up in the sample exposed to TT material; vinculin and integrin signals appear stronger at 24 h follow-up in the sample exposed to BIOS material. These data confirmed how dentinal derivates present satisfying biocompatibility and high conductivity and inductivity properties fundamental in the regenerative processes. Furthermore, the knowledge of the effects of the dentin's degree of mineralization on cellular behavior will help clinicians choose the type of dentine derivates material according to the required clinical situation.


Properties of glycerol-75Se-triether: A lipid-soluble marker for the estimation of intestinal fat absorption.
The properties of a 75Se-labeled glycerol triether were investigated in rat experiments designed to test this substance as a nonabsorbable marker for the assessment of intestinal fat absorption. After oral administration of 75Se-triether, the radioactivity was excreted almost completely with the feces. Amounts in excess of the quantity required tor clinical use did not interfere with overall fat absorption. No evidence for toxicity of 75Se-triether was observed. 131l-triolein was used as tracer fat and fat absorption was calculated by the following methods: (1) isotope balance method-oral intake minus fecal excretion of 131L; (2) isotope ratio method-comparison of the 131L to 75Se ratios in the test dose and in a stool sample. Results obtained from the isotope ratio method were in close agreement with those of the isotope balance method over a range of fat absorption of 80 to 95 per cent, thus indicating that the marker and the radioactive fat pass the gastrointestinal tract at the same rate under these experimental conditions. These results show that 75Se-triether possesses several of the properties of an ideal marker for fat absorption studies. Its advantages over other proposed markers for fat absorption studies are discussed. Simultaneous administration of 131L-TRIOLEIN AND 75Se-triether in a single dose may provide a reliable, rapid, and simple method to estimate intestinal fat absorption in man.


MI-Environment: Geospatial patterns and inequality of relative heat stress vulnerability in Michigan.
Heat stress causes morbidity and mortality and is increasing with climate change. Heat stress can pose particular challenges in northern regions not well adapted to heat. To assist decision makers, we identified the relative vulnerability of census tracts within Michigan to factors that increase exposure to heat stress or reflect susceptibilities in the population based on a California heat vulnerability index. In the MI-Environment assessment, we used a Geographic Information System (GIS) to combine future ensemble climate model projections to create a total of 9 geospatial and demographic variables. As part of a broader planned cumulative environmental exposure assessment, the statewide heat vulnerability index (HVI) maps display the location and relative magnitude of exposure on three metrics: built environment (Place), future expected long-term temperature averages (Temperature), and population susceptibility (People). We observed varied and distinct patterns for each of the three component indices. We assessed how equitably those exposures are distributed by racial and socioeconomic factors. This analysis showed that each of the component indices and the aggregate HVI are disproportionately distributed along racial and socioeconomic lines in Michigan. Census tracts with higher percentages of people of color had larger exposure to HVI factors with a deviation from equity of -0.115 [95% CI -0.108, -0.122]. Similarly, for census tracts with higher percentage of people experiencing poverty, the deviation from equity was -0.101 [95% CI -0.094, -0.107]. The MI-Environment visualization tool can help communities prepare for climate change and resolve inequities by identifying census tracts with the most vulnerable residents and highest potential exposures.


Identification and fine-mapping of QYrAS286-2BL conferring adult-plant resistance to stripe rust in cultivated emmer wheat.
A novel major adult-plant stripe rust resistance QTL derived from cultivated emmer wheat was mapped to a 123.6-kb region on wheat chromosome 2BL. Stripe rust, caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is one of the most devastating diseases of wheat. Identification of new sources of resistance and their utilization in breeding programs is the effectively control strategy. The objective of this study was to identify and genetically characterize the stripe rust resistance derived from the cultivated emmer accession AS286. A recombinant inbred line population, developed from a cross between the susceptible durum wheat line langdon and AS286, was genotyped using the Wheat55K single nucleotide polymorphism array and evaluated in field conditions with a mixture of the prevalent Chinese Pst races (CYR32, CYR33, CYR34, Zhong4, and HY46) and in growth chamber with race CYR34. Three QTLs conferring resistance were mapped on chromosomes 1BS, 2BL, and 5BL, respectively. The QYrAS286-1BS and QYrAS286-2BL were stable with major effects, explaining 12.91% to 18.82% and 11.31% to 31.43% of phenotypic variation, respectively. QYrAS286-5BL was only detected based on growth chamber seedling data. RILs harboring both QYrAS286-1BS and QYrAS286-2BL showed high levels of stripe rust resistance equal to the parent AS286. The QYrAS286-2BL was only detected at the adult-plant stage, which is different from previously named Yr genes and inherited as a single gene. It was further mapped to a 123.6-kb region using KASP markers derived from SNPs identified by bulked segregant RNA sequencing (BSR-Seq). The identified loci enrich our stripe rust resistance gene pool, and the flanking markers developed here could be useful in marker-assisted selection for incorporating QYrAS286-2BL into wheat cultivars.


Integrated mental health atlas of the Western Sydney Local Health District: gaps and recommendations.
Objective Australian mental health care remains hospital centric and fragmented; it is riddled with gaps and does little to promote recovery. Reform must be built on better knowledge of the shape of existing services. Mental health atlases are an essential part of this knowledge base, enabling comparison with other regions and jurisdictions, but must be based on a rigorous classification of services. The main aim of this study is to create an integrated mental health atlas of the Western Sydney LHD in order to help decision makers to better plan informed by local evidence. Methods The standard classification system, namely the Description and Evaluation of Services and Directories in Europe for Long-term Care model, was used to describe and classify adult mental health services in the Western Sydney Local Health District (LHD). This information provided the foundation for accessibility maps and the analysis of the provision of care for people with a lived experience of mental illness in Western Sydney LHD. All this data was used to create the Integrated Mental Health Atlas of Western Sydney LHD. Results The atlas identified four major gaps in mental health care in Western Sydney LHD: (1) a lack of acute and sub-acute community residential care; (2) an absence of services providing acute day care and non-acute day care; (3) low availability of specific employment services for people with a lived experience of mental ill-health; and (4) a lack of comprehensive data on the availability of supported housing. Conclusions The integrated mental health atlas of the Western Sydney LHD provides a tool for evidence-informed planning and critical analysis of the pattern of adult mental health care. What is known about the topic? Several reports have highlighted that the Australian mental health system is hospital based and fragmented. However, this knowledge has had little effect on actually changing the system. What does this paper add? This paper provides a critical analysis of the pattern of adult mental health care provided within the boundaries of the Western Sydney LHD using a standard, internationally validated tool to describe and classify the services. This provides a good picture of the availability of adult mental health care at the local level that was hitherto lacking. What are the implications for practitioners? The data presented herein provide a better understanding of the context in which mental health practitioners work. Managers and planners of services providing care for people with a lived experience of mental illness can use the information herein for better planning informed by local evidence.


Cell types, network homeostasis, and pathological compensation from a biologically plausible ion channel expression model.
How do neurons develop, control, and maintain their electrical signaling properties in spite of ongoing protein turnover and perturbations to activity? From generic assumptions about the molecular biology underlying channel expression, we derive a simple model and show how it encodes an "activity set point" in single neurons. The model generates diverse self-regulating cell types and relates correlations in conductance expression observed in vivo to underlying channel expression rates. Synaptic as well as intrinsic conductances can be regulated to make a self-assembling central pattern generator network; thus, network-level homeostasis can emerge from cell-autonomous regulation rules. Finally, we demonstrate that the outcome of homeostatic regulation depends on the complement of ion channels expressed in cells: in some cases, loss of specific ion channels can be compensated; in others, the homeostatic mechanism itself causes pathological loss of function.


SINCERITIES: inferring gene regulatory networks from time-stamped single cell transcriptional expression profiles.
Single cell transcriptional profiling opens up a new avenue in studying the functional role of cell-to-cell variability in physiological processes. The analysis of single cell expression profiles creates new challenges due to the distributive nature of the data and the stochastic dynamics of gene transcription process. The reconstruction of gene regulatory networks (GRNs) using single cell transcriptional profiles is particularly challenging, especially when directed gene-gene relationships are desired. We developed SINCERITIES (SINgle CEll Regularized Inference using TIme-stamped Expression profileS) for the inference of GRNs from single cell transcriptional profiles. We focused on time-stamped cross-sectional expression data, commonly generated from transcriptional profiling of single cells collected at multiple time points after cell stimulation. SINCERITIES recovers directed regulatory relationships among genes by employing regularized linear regression (ridge regression), using temporal changes in the distributions of gene expressions. Meanwhile, the modes of the gene regulations (activation and repression) come from partial correlation analyses between pairs of genes. We demonstrated the efficacy of SINCERITIES in inferring GRNs using in silico time-stamped single cell expression data and single cell transcriptional profiles of THP-1 monocytic human leukemia cells. The case studies showed that SINCERITIES could provide accurate GRN predictions, significantly better than other GRN inference algorithms such as TSNI, GENIE3 and JUMP3. Moreover, SINCERITIES has a low computational complexity and is amenable to problems of extremely large dimensionality. Finally, an application of SINCERITIES to single cell expression data of T2EC chicken erythrocytes pointed to BATF as a candidate novel regulator of erythroid development. MATLAB and R version of SINCERITIES are freely available from the following websites: http://www.cabsel.ethz.ch/tools/sincerities.html and https://github.com/CABSEL/SINCERITIES. The single cell THP-1 and T2EC transcriptional profiles are available from the original publications (Kouno et al., 2013; Richard et al., 2016). The in silico single cell data are available on SINCERITIES websites. Supplementary data are available at Bioinformatics online.


Stroma-derived HGF drives metabolic adaptation of colorectal cancer to angiogenesis inhibitors.
The role of paracrine Hepatocyte Growth Factor (HGF) in the resistance to angiogenesis inhibitors (AIs) is hidden in xenograft models because mouse HGF fails to fully activate human MET. To uncover it, we compared the efficacy of AIs in wild-type and human HGF knock-in SCID mice bearing orthotopic human colorectal tumors. Species-specific HGF/MET signaling dramatically impaired the response to anti-angiogenic agents and boosted metastatic dissemination. In cell-based assays mimicking the consequences of anti-angiogenic therapy, colorectal cancer cells were completely resistant to hypoxia but extremely sensitive to nutrient deprivation. Starvation-induced apoptosis could be prevented by HGF, which promoted GLUT1-mediated glucose uptake, sustained glycolysis and activated autophagy. Pharmacological inhibition of GLUT1 in the presence of glucose killed tumor cells as effectively as glucose deprivation, and this effect was antagonized by HGF. Concomitant targeting of GLUT1 and HGF potently suppressed growth and dissemination of AI-resistant human tumors in human HGF knock-in SCID mice without exacerbating tumor hypoxia. These data suggest that stroma-derived HGF protects CRC cells against glucose starvation-induced apoptosis, promoting resistance to both AIs and anti-glycolytic agents. Combined inhibition of glucose metabolism and HGF/MET signaling ('anti-METabolic therapy') may represent a more effective CRC treatment compared to utterly blocking tumor blood supply.


Orderly and nonstochastic acquisition of CD94/NKG2 receptors by developing NK cells derived from embryonic stem cells in vitro.
In mice there are two families of MHC class I-specific receptors, namely the Ly49 and CD94/NKG2 receptors. The latter receptors recognize the nonclassical MHC class I Qa-1(b) and are thought to be responsible for the recognition of missing-self and the maintenance of self-tolerance of fetal and neonatal NK cells that do not express Ly49. Currently, how NK cells acquire individual CD94/NKG2 receptors during their development is not known. In this study, we have established a multistep culture method to induce differentiation of embryonic stem (ES) cells into the NK cell lineage and examined the acquisition of CD94/NKG2 by NK cells as they differentiate from ES cells in vitro. ES-derived NK (ES-NK) cells express NK cell-associated proteins and they kill certain tumor cell lines as well as MHC class I-deficient lymphoblasts. They express CD94/NKG2 heterodimers, but not Ly49 molecules, and their cytotoxicity is inhibited by Qa-1(b) on target cells. Using RT-PCR analysis, we also report that the acquisition of these individual receptor gene expressions during different stages of differentiation from ES cells to NK cells follows a predetermined order, with their order of acquisition being first CD94; subsequently NKG2D, NKG2A, and NKG2E; and finally, NKG2C. Single-cell RT-PCR showed coexpression of CD94 and NKG2 genes in most ES-NK cells, and flow cytometric analysis also detected CD94/NKG2 on most ES-NK cells, suggesting that the acquisition of these receptors by ES-NK cells in vitro is nonstochastic, orderly, and cumulative.


Chaperones rescue luciferase folding by separating its domains.
Over the last 50 years, significant progress has been made toward understanding how small single-domain proteins fold. However, very little is known about folding mechanisms of medium and large multidomain proteins that predominate the proteomes of all forms of life. Large proteins frequently fold cotranslationally and/or require chaperones. Firefly (Photinus pyralis) luciferase (Luciferase, 550 residues) has been a model of a cotranslationally folding protein whose extremely slow refolding (approximately days) is catalyzed by chaperones. However, the mechanism by which Luciferase misfolds and how chaperones assist Luciferase refolding remains unknown. Here we combine single-molecule force spectroscopy (atomic force microscopy (AFM)/single-molecule force spectroscopy) with steered molecular dynamic computer simulations to unravel the mechanism of chaperone-assisted Luciferase refolding. Our AFM and steered molecular dynamic results show that partially unfolded Luciferase, with the N-terminal domain remaining folded, can refold robustly without chaperones. Complete unfolding causes Luciferase to get trapped in very stable non-native configurations involving interactions between N- and C-terminal residues. However, chaperones allow the completely unfolded Luciferase to refold quickly in AFM experiments, strongly suggesting that chaperones are able to sequester non-natively contacting residues. More generally, we suggest that many chaperones, rather than actively promoting the folding, mimic the ribosomal exit tunnel and physically separate protein domains, allowing them to fold in a cotranslational-like sequential process.


Reflections on an unsolved problem of biology: the evolution of senescence and death.
The evolutionary theory of senescence is based largely on principles first outlined by Williams in 1957, and consists of two relatively independent parts. The first part builds on ideas first put forward by Medawar, Haldane and others, to explain how something as negative as senescence could have been positively selected in evolution, particularly since most animals in the wild do not reach an age where senescence is expressed. Williams proposed that the genes responsible for the negative effects of senecence (senescence effector genes) were fixed in evolution by a process he called antagonistic pleiotropy, wherein a subset of genes selected because they confer a reproductive advantage early in life may have harmful effects in the post-reproductive period; negative selection against these harmful effects fails because, as pointed out by Medawar, the force of natural selection declines with age. The evolutionary history of senescence-causing genes is seen as a nondirected accumulation of genes selected on a basis independent of senescence per se. In the second portion of his paper, Williams made a series of predictions about how the age of organisms at reproductive maturity, fecundity, lifespan and the timing of the onset of senescence would all interact in the life history of a species. These latter predictions, which do not depend at all on details of the mechanisms of selection of senescence effector genes, have been validated by numerous experiments over the past several decades. On the other hand, it has become increasingly evident that the senescence effector genes did not, as would be predicted by antagonistic pleiotropy, accumulate in a random, non-directed fashion in various species over evolutionary time. Rather, everything we know about these genes suggests they were present in eukaryotic founder cells shortly after, or even congruent with, the emergence of eukaryotes from their prokaryotic ancestors, and have been stringently conserved ever since. Complicated explanations of how so-called "death genes" may have evolved in eukaryotes are thus not required. It is suggested that the evolutionary theory of senescence should be focused on those evolutionary principles that have been validated experimentally, and that the notion of antagonistic pleiotropy--which cannot be experimentally validated--be dropped from our thinking about the evolution of senescence.


Novel insights into the unfolded protein response using Pichia pastoris specific DNA microarrays.
DNA Microarrays are regarded as a valuable tool for basic and applied research in microbiology. However, for many industrially important microorganisms the lack of commercially available microarrays still hampers physiological research. Exemplarily, our understanding of protein folding and secretion in the yeast Pichia pastoris is presently widely dependent on conclusions drawn from analogies to Saccharomyces cerevisiae. To close this gap for a yeast species employed for its high capacity to produce heterologous proteins, we developed full genome DNA microarrays for P. pastoris and analyzed the unfolded protein response (UPR) in this yeast species, as compared to S. cerevisiae. By combining the partially annotated gene list of P. pastoris with de novo gene finding a list of putative open reading frames was generated for which an oligonucleotide probe set was designed using the probe design tool TherMODO (a thermodynamic model-based oligoset design optimizer). To evaluate the performance of the novel array design, microarrays carrying the oligo set were hybridized with samples from treatments with dithiothreitol (DTT) or a strain overexpressing the UPR transcription factor HAC1, both compared with a wild type strain in normal medium as untreated control. DTT treatment was compared with literature data for S. cerevisiae, and revealed similarities, but also important differences between the two yeast species. Overexpression of HAC1, the most direct control for UPR genes, resulted in significant new understanding of this important regulatory pathway in P. pastoris, and generally in yeasts. The differences observed between P. pastoris and S. cerevisiae underline the importance of DNA microarrays for industrial production strains. P. pastoris reacts to DTT treatment mainly by the regulation of genes related to chemical stimulus, electron transport and respiration, while the overexpression of HAC1 induced many genes involved in translation, ribosome biogenesis, and organelle biosynthesis, indicating that the regulatory events triggered by DTT treatment only partially overlap with the reactions to overexpression of HAC1. The high reproducibility of the results achieved with two different oligo sets is a good indication for their robustness, and underlines the importance of less stringent selection of regulated features, in order to avoid a large number of false negative results.


The future of fish in Africa: Employment and investment opportunities.
One of the most pressing challenges facing food systems in Africa is ensuring availability of a healthy and sustainable diet to 2.4 billion people by 2050. The continent has struggled with development challenges, particularly chronic food insecurity and pervasive poverty. In Africa's food systems, fish and other aquatic foods play a multifaceted role in generating income, and providing a critical source of essential micronutrients. To date, there are no estimates of investment and potential returns for domestic fish production in Africa. To contribute to policy debates about the future of fish in Africa, we applied the International Model for Policy Analysis of Agriculture Commodities and Trade (IMPACT) to explore two Pan-African scenarios for fish sector growth: a business-as-usual (BAU) scenario and a high-growth scenario for capture fisheries and aquaculture with accompanying strong gross domestic product growth (HIGH). Post-model analysis was used to estimate employment and aquaculture investment requirements for the sector in Africa. Africa's fish sector is estimated to support 20.7 million jobs in 2030, and 21.6 million by 2050 under the BAU. Approximately 2.6 people will be employed indirectly along fisheries and aquaculture value chains for every person directly employed in the fish production stage. Under the HIGH scenario, total employment in Africa's fish food system will reach 58.0 million jobs, representing 2.4% of total projected population in Africa by 2050. Aquaculture production value is estimated to achieve US$ 3.3 billion and US$ 20.4 billion per year under the BAU and HIGH scenarios by 2050, respectively. Farm-gate investment costs for the three key inputs (fish feeds, farm labor, and fish seed) to achieve the aquaculture volumes projected by 2050 are estimated at US$ 1.8 billion per year under the BAU and US$ 11.6 billion per year under the HIGH scenario. Sustained investments are critical to sustain capture fisheries and support aquaculture growth for food system transformation towards healthier diets.


Effect of acute compartmental pressure change on response to vibratory stimuli in primates.
This study investigated the possibility that a vibratory stimulus could discriminate the effect of relatively low pressures on nerve function in an acute compartmental syndrome in a primate model. The first phase of the study utilized somatosensory-evoked potentials to determine the outcome of varying pressures in an acute carpal tunnel syndrome in the anesthetized monkey. For increasing carpal tunnel pressures above 30 mmHg, the amplitude of the compound action potential of the A beta wave (touch fiber) generated in the median nerve by an electric stimulus to the index finger progressively decreased to a complete conduction block. It took progressively less time to achieve the conduction block at higher compartmental pressures. When compartmental pressure was released, the time required for the conduction block to return to normal was in direct relationship to the time required to complete the block. For carpal tunnel pressure between 15 and 30 mmHg, an increase in the height of this compound action potential was observed. The second phase of the study utilized the perceptual judgments of an awake monkey trained to discriminate differences in amplitude of a 10-Hz vibratory stimulus to the hairs of the dorsum of the foot. An anterior compartment pressure of 37 mmHg for 1 1/2 hours significantly decreased the monkey's ability to discriminate between the two amplitudes. At 50 mmHg in the anterior compartment, ability to discriminate between vibratory stimuli was further impaired. This study supports the use of noninvasive vibratory stimuli, such as a tuning fork, to evaluate acute compartmental syndromes.


Bioassays for the characterisation and control of therapeutic cytokines; determination of potency.
The primary value of bioassays is that they alone directly assess the biological activity of bioactive substances and products like cytokines. Appropriately designed bioassays reflect the fundamental aspects of the biological activity of a cytokine molecule, including ligand-receptor binding, signal transduction processes (often poorly understood) and the final observed biological effects. Biological assays therefore complement physicochemical and biochemical procedures which normally only assess precise molecular structural features of cytokines. Bioassays provide valuable information concerning the potency of cytokine products. This is essential for evaluating batch-to-batch consistency, appropriate formulations and stability. Bioassay data are crucial at all stages in the development of cytokine products, from early research to final quality control of finished product. However, the type and design of bioassays may differ according to the information required and its intended use. The assays may or may not directly relate to the clinical use of the product. Bioassays can be difficult to perform and time consuming, although this often reflects bad assay choice and/or design. Correct analysis of the assay results is essential if valid data are to be obtained. Standardisation, using correctly calibrated primary and secondary standards, is essential. In vivo bioassays are normally more unreliable than in vitro procedures, but in some cases in vitro systems are either not available or do not address important biological characteristics of a product. Bioassays must be validated for their intended purpose and for the types of samples to be measured. Appropriate statistical analysis should be used to derive the significance and specifications of results. This needs to address both variability in samples and assay performance. Specifications (limits) for product acceptability need to be derived from real data using several batches of cytokine product.


Clinical types of proarrhythmic response to antiarrhythmic drugs.
The problem of drug-induced or drug-aggravated cardiac arrhythmias has been recognized for many years. Digitalis glycosides and class I, II, III, or IV antiarrhythmic drugs can cause severe sinus node dysfunction or atrioventricular block. Digitalis can cause a variety of supraventricular arrhythmias; atrial tachycardia with block and nonparoxysmal atrioventricular junctional tachycardia are the most characteristic. Recognition that class IA and class III antiarrhythmic drugs can aggravate arrhythmias or cause new arrhythmias in patients being treated for potentially malignant or malignant ventricular arrhythmias has intensified the interest in proarrhythmia in recent years. Several characteristic types of proarrhythmic response have been described. Torsades de pointes (multiform) ventricular tachycardia (VT) accompanied by prolongation of the QT interval can be caused by class IA and class III antiarrhythmic drugs as well as other drugs that bind to the membrane sodium channels of ventricular cells, for example, tricyclic antidepressants. Uniform VT in patients with malignant ventricular arrhythmias and poor left ventricular function is a characteristic proarrhythmic response to class IC antiarrhythmic drugs. Bidirectional VT or accelerated idioventricular rhythm are characteristic of digitalis toxicity. More difficult to establish as proarrhythmic responses are increased frequency of ventricular premature depolarizations and increased ease of inducing VT with programmed ventricular stimulation.


The head inducer Cerberus is a multifunctional antagonist of Nodal, BMP and Wnt signals.
Embryological and genetic evidence indicates that the vertebrate head is induced by a different set of signals from those that organize trunk-tail development. The gene cerberus encodes a secreted protein that is expressed in anterior endoderm and has the unique property of inducing ectopic heads in the absence of trunk structures. Here we show that the cerberus protein functions as a multivalent growth-factor antagonist in the extracellular space: it binds to Nodal, BMP and Wnt proteins via independent sites. The expression of cerberus during gastrulation is activated by earlier nodal-related signals in endoderm and by Spemann-organizer factors that repress signalling by BMP and Wnt. In order for the head territory to form, we propose that signals involved in trunk development, such as those involving BMP, Wnt and Nodal proteins, must be inhibited in rostral regions.


A resurrection study reveals rapid adaptive evolution within populations of an invasive plant.
The future spread and impact of an introduced species will depend on how it adapts to the abiotic and biotic conditions encountered in its new range, so the potential for rapid evolution subsequent to species introduction is a critical, evolutionary dimension of invasion biology. Using a resurrection approach, we provide a direct test for change over time within populations in a species' introduced range, in the Asian shade annual Polygonum cespitosum. We document, over an 11-year period, the evolution of increased reproductive output as well as greater physiological and root-allocational plasticity in response to the more open, sunny conditions found in the North American range in which the species has become invasive. These findings show that extremely rapid adaptive modifications to ecologically-important traits and plastic expression patterns can evolve subsequent to a species' introduction, within populations established in its introduced range. This study is one of the first to directly document evolutionary change in adaptive plasticity. Such rapid evolutionary changes can facilitate the spread of introduced species into novel habitats and hence contribute to their invasive success in a new range. The data also reveal how evolutionary trajectories can differ among populations in ways that can influence invasion dynamics.


Smad3 dosage determines androgen responsiveness and sets the pace of postnatal testis development.
The establishment and maturation of the testicular Sertoli cell population underpins adult male fertility. These events are influenced by hormones and endocrine factors, including FSH, testosterone and activin. Activin A has developmentally regulated effects on Sertoli cells, enhancing proliferation of immature cells and later promoting postmitotic maturation. These differential responses correlate with altered mothers against decapentaplegic (SMAD)-2/3 signaling: immature cells signal via SMAD3, whereas postmitotic cells use both SMAD2 and SMAD3. This study examined the contribution of SMAD3 to postnatal mouse testis development. We show that SMAD3 production and subcellular localization are highly regulated and, through histological and molecular analyses, identify effects of altered Smad3 dosage on Sertoli and germ cell development. Smad3(+/-) and Smad3(-/-) mice had smaller testes at 7 d postpartum, but this was not sustained into adulthood. Juvenile and adult serum FSH levels were unaffected by genotype. Smad3-null mice displayed delayed Sertoli cell maturation and had reduced expression of androgen receptor (AR), androgen-regulated transcripts, and Smad2, whereas germ cell and Leydig cell development were essentially normal. This contrasted remarkably with advanced Sertoli and germ cell maturation and increased expression of AR and androgen-regulated transcripts in Smad3(+/-) mice. In addition, SMAD3 was down-regulated during testis development and testosterone up-regulated Smad2, but not Smad3, in the TM4 Sertoli cell line. Collectively these data reveal that appropriate SMAD3-mediated signaling drives normal Sertoli cell proliferation, androgen responsiveness, and maturation and influences the pace of the first wave of spermatogenesis, providing new clues to causes of altered pubertal development in boys.


Fatal intoxication by 5F-ADB and diphenidine: Detection, quantification, and investigation of their main metabolic pathways in humans by LC/MS/MS and LC/Q-TOFMS.
Despite the implementation of a new blanket scheduling system in 2013, new psychoactive substance (NPS) abuse remains a serious social concern in Japan. We present a fatal intoxication case involving 5F-ADB (methyl 2-[1-(5-fluoropentyl)-1H-indazole-3-carboxamido]-3,3-dimethylbutanoate) and diphenidine. Postmortem blood screening by liquid chromatography/quadrupole time-of-flight mass spectrometry (LC/Q-TOFMS) in the information-dependent acquisition mode only detected diphenidine. Further urinary screening using an in-house database containing NPS and metabolites detected not only diphenidine but also possible 5F-ADB metabolites; subsequent targeted screening by LC/tandem mass spectrometry (LC/MS/MS) allowed for the detection of a very low level of unchanged 5F-ADB in postmortem heart blood. Quantification by standard addition resulted in the postmortem blood concentrations being 0.19 ± 0.04 ng/mL for 5F-ADB and 12 ± 2.6 ng/mL for diphenidine. Investigation of the urinary metabolites revealed pathways involving ester hydrolysis (M1) and oxidative defluorination (M2), and further oxidation to the carboxylic acid (M3) for 5F-ADB. Mono- and di-hydroxylated diphenidine metabolites were also found. The present case demonstrates the importance of urinary metabolite screening for drugs with low blood concentration. Synthetic cannabinoids (SCs) fluorinated at the terminal N-alkyl position are known to show higher cannabinoid receptor affinity relative to their non-fluorinated analogues; 5F-ADB is no exception with high CB1 receptor activity and much greater potency than Δ9 -THC and other earlier SCs, thus we suspect its acute toxicity to be high compared to other structurally related SC analogues. The low blood concentration of 5F-ADB may be attributed to enzymatic and/or non-enzymatic degradation, and further investigation into these possibilities is underway.


Theoretical Effects of Substituting Butter with Margarine on Risk of Cardiovascular Disease.
Several recent articles have called into question the deleterious effects of high animal fat diets due to mixed results from epidemiologic studies and the lack of clinical trial evidence in meta-analyses of dietary intervention trials. We were interested in examining the theoretical effects of substituting plant-based fats from different types of margarine for animal-based fat from butter on the risk of atherosclerosis-related cardiovascular disease (CVD). We prospectively studied 71,410 women, aged 50-79 years, and evaluated their risk for clinical myocardial infarction (MI), total coronary heart disease (CHD), ischemic stroke, and atherosclerosis-related CVD with an average of 13.2 years of follow-up. Butter and margarine intakes were obtained at baseline and year 3 by means of a validated food frequency questionnaire. Cox proportional hazards regression using a cumulative average diet method was used to estimate the theoretical effect of substituting 1 teaspoon/day of three types of margarine for the same amount of butter. Substituting butter or stick margarine with tub margarine was associated with lower risk of MI (HRs = 0.95 and 0.91). Subgroup analyses, which evaluated these substitutions among participants with a single source of spreadable fat, showed stronger associations for MI (HRs = 0.92 and 0.87). Outcomes of total CHD, ischemic stroke, and atherosclerosis-related CVD showed wide confidence intervals but the same trends as the MI results. This theoretical dietary substitution analysis suggests that substituting butter and stick margarine with tub margarine when spreadable fats are eaten may be associated with reduced risk of myocardial infarction.


Seroprevalence of Toxoplasma gondii in donkeys (Equus asinus) in Italy.
Toxoplasmosis, an important zoonosis, can be transmitted by eating meat or drinking milk of animals infected with Toxoplasma gondii. Samples were collected from 238 donkeys in the year 2010 in Italy, which included 207 females and 31 males of five breeds and crossbreeds with the average age 9 years (1 month-24 years). Sera were tested for T. gondii antibodies using a latex agglutination test and the indirect fluorescent antibody test; 5 and 8% seropositivity were recorded, respectively. We found significant correlation between the presence of T. gondii antibodies and sex, age, grazing and presence of cats on the farms and their access to donkey feed. This is the first detection of T. gondii antibodies in donkeys in Italy.


Invasive Haemophilus influenzae disease and epiglottitis among Swiss children from 1980 to 1993: evidence for herd immunity among older age groups. The Swiss Haemophilus Influenzae Study Group.
We analysed time trends in the incidence of invasive Haemophilus influenzae disease in Switzerland between 1980 and 1993 to investigate whether conjugated H. influenzae type b vaccines, licensed in Switzerland in May 1990, confer indirect protection to children in older, non-vaccinated age groups. Data were obtained from the records of 39 Swiss paediatric clinics for 2,857 children 0-16 years old with invasive H. influenzae disease. Incidence time trends were analysed by Poisson regression. The diseases incidence decreased by 80% among 0-4-year-old children (i.e. those eligible for vaccination) between 1990 and 1993. Among children aged 5-16 years, there also was an abrupt 50% fall in the incidence of H. influenzae meningitis after 1990. However, the incidence of epiglottitis in this age group had started to decline before conjugated vaccines became available, with no additional decline thereafter. While our results suggest some indirect protection conferred to older, non-vaccinated children through the administration of conjugated vaccines to younger children, they also argue that underlying time trends of invasive H. influenzae disease need not be considered when interpreting incidence rates in the vaccine era.


Population genomics of an outbreak of the potato late blight pathogen, Phytophthora infestans, reveals both clonality and high genotypic diversity.
An outbreak of the potato late blight pathogen Phytophthora infestans in Denmark was characterized in order to resolve the population structure and determine to what extent sexual reproduction was occurring. A standard set of microsatellite simple sequence repeats (SSRs) and single nucleotide polymorphism (SNP) markers generated using restriction site-associated DNA sequencing (RAD-seq) were employed in parallel. A total of 83 individuals, isolated from seven different potato fields in 2014, were analysed together with five Danish whole-genome sequenced isolates, as well as two Mexican individuals used as an outgroup. From a filtered dataset of 55 288 SNPs, population genomics analyses revealed no sign of recombination, implying clonality. In spite of this, multilocus genotypes were unique to individual potato fields, with little evidence of gene flow between fields. Ploidy analysis performed on the SNPs dataset indicated that the majority of isolates were diploid. These contradictory results with clonality and high genotypic diversity may suggest that rare sexual events likely still contribute to the population. Comparison of the results generated by SSRs vs SNPs data indicated that large marker sets, generated by RAD-seq, may be advised going forward, as it provides a higher level of genetic discrimination than SSRs.


Cross-Sectional Study to Determine the Prevalence of Hepatitis B and C Virus Infection in High Risk Groups in the Northeast Region of Brazil.
Background: HBV (Hepatitis B Virus) and HCV (Hepatitis C Virus) infections are more prevalent in vulnerable populations than the general population. The objective of this study was to investigate the prevalence of HBV and HCV infection in HIV-positive patients (GI), chronic renal failure (CRF) patients (GII) and coagulation disorder individuals (GIII). Methods: A cross-sectional study was conducted from June 2014 to March 2015. Serum samples were tested for markers of hepatitis B and C by enzyme-linked immunosorbent assay (ELISA). Sociodemographic, epidemiological, clinical and laboratory data and accompanying statistical analyses were performed using Epi Info™ 7. Results: A total of 348 individuals were recruited, i.e., 154 HIV-positive, 143 CRF and 51 coagulopathy patients. Among them, more than 66% were men, and the predominant age group was 26-35 years in GI and 56-65 years in GIII. Most patients had more than 8 years of education (66.2% in GI, 60.6% in GIII and 46.1% in GII), with a family income between 100-400 dollars in more than 48% of patients. The prevalence of the HBsAg marker was 3.9%, 7% and 3.9%, total anti-HBc was 28.6%, 55.9% and 31.4%, and anti-HCV was 1.3%, 12.6% and 47% for GI, GII and GIII, respectively. However, the prevalence of anti-HBs was greater than 70% in all groups. Conclusions: This study shows a high prevalence of HBV and HCV among specific groups compared to the general population. Factors such as age, income, number of sexual partners, sexually transmitted disease burden, blood transfusion history or blood products and blood transfusions before 1994 were associated with a higher prevalence for these infections.


Validation of 3D ultra-short TE (UTE) Phase-Contrast MRI for imaging of steady flow: initial phantom experiments.
Assessment of blood flow is an important factor in diagnosis of cardiovascular disease. Vascular stenosis result in disturbed blood flow, flow recirculation, turbulence, and flow jet. These types of flows cause erroneous quantification of blood flow using conventional Phase contrast (PC) MRI techniques. Previous investigations have revealed that shorter Echo Times (TE) can decrease the quantification errors. In this paper, we performed phantom studies under steady flow to validate the UTE technique. Investigation of three different constant flow rates revealed a significant improvement in flow quantification and reduction of flow artifacts in comparison to Cartesian Phase-Contrast MRI.


A general framework for estimating volume-outcome associations from longitudinal data.
Recently, there has been much interest in using volume-outcome data to establish causal associations between measures of surgical experience or quality and patient outcomes following a surgical procedure, such as coronary artery bypass graft, total hip replacement, and radical prostatectomy. However, there does not appear to be a standard approach to a volume-outcome analysis with respect to specifying a volume measure and selecting an estimation method. We establish the recurrent marked point process as a general framework from which to approach a longitudinal volume-outcome analysis and examine the statistical issues associated with using longitudinal data analysis methods to model aggregate volume-outcome data. We review assumptions to ensure that linear or generalized linear mixed models and generalized estimating equations provide valid estimates of the volume-outcome association. In addition, we provide theoretical and empirical evidence that bias may be introduced when an aggregate volume measure is used to address a scientific question regarding the effect of cumulative experience. We conclude with the recommendation that analysts carefully specify a volume measure that most accurately reflects their scientific question of interest and select an estimation method that is appropriate for their scientific context.


Genetics of decayed sexual traits in a parasitoid wasp with endosymbiont-induced asexuality.
Trait decay may occur when selective pressures shift, owing to changes in environment or life style, rendering formerly adaptive traits non-functional or even maladaptive. It remains largely unknown if such decay would stem from multiple mutations with small effects or rather involve few loci with major phenotypic effects. Here, we investigate the decay of female sexual traits, and the genetic causes thereof, in a transition from haplodiploid sexual reproduction to endosymbiont-induced asexual reproduction in the parasitoid wasp Asobara japonica. We take advantage of the fact that asexual females cured of their endosymbionts produce sons instead of daughters, and that these sons can be crossed with sexual females. By combining behavioral experiments with crosses designed to introgress alleles from the asexual into the sexual genome, we found that sexual attractiveness, mating, egg fertilization and plastic adjustment of offspring sex ratio (in response to variation in local mate competition) are decayed in asexual A. japonica females. Furthermore, introgression experiments revealed that the propensity for cured asexual females to produce only sons (because of decayed sexual attractiveness, mating behavior and/or egg fertilization) is likely caused by recessive genetic effects at a single locus. Recessive effects were also found to cause decay of plastic sex-ratio adjustment under variable levels of local mate competition. Our results suggest that few recessive mutations drive decay of female sexual traits, at least in asexual species deriving from haplodiploid sexual ancestors.


Announcing Swine Flu and the Interpretation of Pandemic Anxiety.
This paper discusses the ways in which 2009 novel swine-origin influenza A (H1N1) was announced and resonated with current pandemic anxieties. In particular, the US Centers for Disease Control and Prevention (CDC) are used as a lens through which recent pandemic anxieties can be analysed and understood. This entails a closer look at the securitisation of public health and the challenges and struggles this may have caused within public health agencies. In that light, CDC' formal entanglement with global health security and its announcement of the H1N1 pandemic are interpreted, followed by an ethnographically informed focus on various people who were engaged in the H1N1 emergency response and their practices and practical struggles in the face of pandemic anxiety.


Measurement of human surfactant protein-B turnover in vivo from tracheal aspirates using targeted proteomics.
We describe a method to measure protein synthesis and catabolism in humans without prior purification and use the method to measure the turnover of surfactant protein-B (SP-B). SP-B, a lung-specific, hydrophobic protein essential for fetal-neonatal respiratory transition, is present in only picomolar quantities in tracheal aspirate samples and difficult to isolate for dynamic turnover studies using traditional in vivo tracer techniques. Using infusion of [5,5,5-(2)H(3)] leucine and a targeted proteomics method, we measured both the quantity and kinetics of SP-B tryptic peptides in tracheal aspirate samples of symptomatic newborn infants. The fractional synthetic rate (FSR) of SP-B measured using the most abundant proteolytic fragment, a 10 amino acid peptide from the carboxy-terminus of proSP-B (SPTGEWLPR), from the circulating leucine pool was 0.035 +/- 0.005 h(-1), and the fractional catabolic rate was 0.044 +/- 0.003 h(-1). This technique permits high-throughput and sensitive measurement of turnover of low abundance proteins with minimal sample preparation.


Physical transfer of membrane and cytoplasmic components as a general mechanism of cell-cell communication.
Recent evidence from different research areas has revealed a novel mechanism of cell-cell communication by spontaneous intercellular transfer of cellular components (ICT). Here we studied this phenomenon by co-culturing different cells that contain distinct levels of proteins or markers for the plasma membrane or cytoplasm. We found that a variety of transmembrane proteins are transferable between multiple cell types. Membrane lipids also show a high efficiency of intercellular transfer. Size-dependent cytoplasmic transfer allows exchange of cytoplasmic macromolecules up to 40 kDa between somatic cells, and up to 2000 kDa between uncommitted human precursor cells and human umbilical vein endothelial cells. Protein transfer, lipid transfer and cytoplasmic component transfer can occur simultaneously and all require direct cell-cell contact. Analyses of the properties of ICT, together with a close examination of cell-cell interactions, suggest that the spontaneous ICT of different cellular components might have a common underlying process: transient local membrane fusions formed when neighboring cells undergo close cell-cell contact.