Transmission of Panulirus argus virus 1 (PaV1) and its effect on the survival of juvenile Caribbean spiny lobster.
The Caribbean spiny lobster Panulirus argus, an important fisheries species, is host to Panulirus argus virus 1 (PaV1), a lethal, unclassified virus--the first found in any species of lobster--prevalent in juvenile lobsters. We describe a series of laboratory experiments aimed at assessing the likely modes of disease transmission, determining the survival of lobsters relative to each transmission pathway and identifying potential alternate hosts. Given evidence for lower prevalence of PaV1 in large lobsters, the effect of lobster size on susceptibility was also examined. Results demonstrated that PaV1 can be transmitted to juvenile lobsters via inoculation, ingestion of diseased tissue, contact with diseased lobsters and--among the smallest juveniles--through water over distances of a few meters. Contact and waterborne transmission, the most likely modes of transmission in the wild, were less efficient than inoculation or ingestion. Nevertheless, about half of the smallest lobsters in contact and waterborne trials contracted the disease and died within 3 mo. Other decapods that co-occur with P. argus (e.g. spotted lobster P. guttatus, stone crab Menippe mercenaria, channel crab Mithrax spinosissimus) did not acquire the disease after inoculation with PaV1-infected hemolymph. Our results confirmed that PaV1 is highly infectious and lethal to juvenile P. argus, particularly early benthic juveniles in the wild, and, hence, is a threat to mariculture.


Celiac disease as a model for the evolution of multifactorial disease in humans.
Celiac disease (CD) is a multifactorial chronic inflammatory condition that results in injury of the mucosal lining of the small intestine upon ingestion of wheat gluten and related proteins from barley and rye. Although the exact mechanisms leading to CD are not fully understood, the genetic basis of CD has been relatively well characterized. In this review we briefly review the history of discovery, clinical presentation, pathophysiology, and current understanding of the genetics underlying CD risk. Then, we discuss what is known about the current distribution and evolutionary history of genes underlying CD risk in light of other evolutionary models of disease. Specifically, we conclude that the set of loci underlying CD risk did not cohesively evolve as a response to a single past selection event such as the development of agriculture. Rather, deterministic and stochastic evolutionary processes have both contributed to the present distribution of variation in CD risk loci. Selection has shaped some components of this network, but this selection appears to have occurred at different points in the past. Other parts of the CD risk network have likely arisen due to stochastic processes such as genetic drift.


Allocation of attentional resources in posthypnotic suggestion.
Highly hypnotizable subjects received a nonhypnotic instruction to respond to a particular digit in a display and a posthypnotic suggestion to respond to a different digit. On some test trials, these 2 responses were tested separately; on others, they were placed in conflict. Overall, subjects were no more responsive to posthypnotic cues than to nonhypnotic cues, nor did their response latencies differ. However, response to posthypnotic cues diminished when they conflicted with the nonhypnotic cues. Analysis of response latencies showed that posthypnotic responding interfered with nonhypnotic responding (and vice versa), even on those trials where there was no procedural conflict. Posthypnotic behavior is not inevitably evoked by the presentation of the prearranged cue. Furthermore, the interference between posthypnotic and nonhypnotic responses indicates that posthypnotic responding consumes attentional resources. Both findings indicate that posthypnotic behavior is not automatic in the technical sense of that term.


Contribution of epididymal epithelial cell functions to sperm epigenetic changes and the health of progeny.
Spermatozoa acquire their motility and fertilizing abilities during their maturation through the epididymis. This process is controlled by epididymal epithelial cells that possess features adapted to sense and respond to their surrounding environment and to communicate with spermatozoa. During the past decade, new intercellular communication processes have been discovered, including the secretion and transport of molecules from the epithelium to spermatozoa via extracellular vesicles (EVs), as well as sensing of the intraluminal milieu by cellular extensions. This review addresses recent findings regarding epididymal epithelial cell features and interactions between spermatozoa and the epididymal epithelium as well as epigenetic modifications undergone by spermatozoa during transit through the epididymal microenvironment. A systematic search was conducted in Pubmed with the keyword 'epididymis'. Results were filtered on original research articles published from 2009 to 2021 and written in the English language. One hundred fifteen original articles presenting recent advancements on the epididymis contribution to sperm maturation were selected. Some additional papers cited in the primary reference were also included. A special focus was given to higher mammalian species, particularly rodents, bovines and humans, that are the most studied in this field. This review provides novel insights into the contribution of epididymal epithelium and EVs to post-testicular sperm maturation. First, new immune cell populations have been described in the epididymis, where they are proposed to play a role in protecting the environment surrounding sperm against infections or autoimmune responses. Second, novel epididymal cell extensions, including dendrites, axopodia and primary cilia, have been identified as sensors of the environment surrounding sperm. Third, new functions have been outlined for epididymal EVs, which modify the sperm epigenetic profile and participate in transgenerational epigenetic inheritance of paternal traits. Although the majority of these findings result from studies in rodents, this fundamental research will ultimately improve our knowledge of human reproductive physiopathologies. Recent discoveries linking sperm epigenetic modifications with paternal environmental exposure and progeny outcome further stress the importance of advancing fundamental research on the epididymis. From this, new therapeutic options for infertile couples and better counseling strategies may arise to increase positive health outcomes in children conceived either naturally or with ART.


TECRL, a new life-threatening inherited arrhythmia gene associated with overlapping clinical features of both LQTS and CPVT.
Genetic causes of many familial arrhythmia syndromes remain elusive. In this study, whole-exome sequencing (WES) was carried out on patients from three different families that presented with life-threatening arrhythmias and high risk of sudden cardiac death (SCD). Two French Canadian probands carried identical homozygous rare variant in TECRL gene (p.Arg196Gln), which encodes the trans-2,3-enoyl-CoA reductase-like protein. Both patients had cardiac arrest, stress-induced atrial and ventricular tachycardia, and QT prolongation on adrenergic stimulation. A third patient from a consanguineous Sudanese family diagnosed with catecholaminergic polymorphic ventricular tachycardia (CPVT) had a homozygous splice site mutation (c.331+1G>A) in TECRL Analysis of intracellular calcium ([Ca2+]i) dynamics in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) generated from this individual (TECRLHom-hiPSCs), his heterozygous but clinically asymptomatic father (TECRLHet-hiPSCs), and a healthy individual (CTRL-hiPSCs) from the same Sudanese family, revealed smaller [Ca2+]i transient amplitudes as well as elevated diastolic [Ca2+]i in TECRLHom-hiPSC-CMs compared with CTRL-hiPSC-CMs. The [Ca2+]i transient also rose markedly slower and contained lower sarcoplasmic reticulum (SR) calcium stores, evidenced by the decreased magnitude of caffeine-induced [Ca2+]i transients. In addition, the decay phase of the [Ca2+]i transient was slower in TECRLHom-hiPSC-CMs due to decreased SERCA and NCX activities. Furthermore, TECRLHom-hiPSC-CMs showed prolonged action potentials (APs) compared with CTRL-hiPSC-CMs. TECRL knockdown in control human embryonic stem cell-derived CMs (hESC-CMs) also resulted in significantly longer APs. Moreover, stimulation by noradrenaline (NA) significantly increased the propensity for triggered activity based on delayed afterdepolarizations (DADs) in TECRLHom-hiPSC-CMs and treatment with flecainide, a class Ic antiarrhythmic drug, significantly reduced the triggered activity in these cells. In summary, we report that mutations in TECRL are associated with inherited arrhythmias characterized by clinical features of both LQTS and CPVT Patient-specific hiPSC-CMs recapitulated salient features of the clinical phenotype and provide a platform for drug screening evidenced by initial identification of flecainide as a potential therapeutic. These findings have implications for diagnosis and treatment of inherited cardiac arrhythmias.


A Performance Improvement Project in Antibiotic Administration for Open Fractures.
To evaluate the effects of a trauma performance improvement project involving standardized protocols for the administration of antibiotics in open fractures at a level one trauma center. This study specifically evaluated the protocol's efficacy for improving the timing of delivery and appropriate therapy administration and sought to identify factors that lead to the delay in antibiotic delivery. Retrospective comparative cohort study comparing patients with open fractures treated at our hospital between January 2013 and September 2015 (group 1) and between April 2016 and June 2017 (group 2). Group 1 was treated before implementation of the performance improvement project and group 2 was treated after implementation. Group 1 consisted of 79 patients and group 2 consisted of 80 patients with open fractures. Each group was statistically similar in patient and injury factors. Group 1 received antibiotics at an average of 97 minutes after arrival to our hospital while group 2 patients received them at an average of 46 minutes (P < 0.0001). Average time from admission to initial evaluation improved from 10 to 3 minutes (P < 0.0001). Average time from evaluation to antibiotic order placement improved from 77 to 26 minutes (P < 0.0001). Average time from order entry to antibiotic administration showed no significant difference (12 versus 15 minutes, P = 0.25). Thirty-four percent (27/79) of group 1 patients and 84% (67/80) of group 2 patients received antibiotics within 1 hour of admission (P < 0.0001), while 91% and 99% received antibiotics within 3 hours, respectively (P = 0.03). The described multifaceted performance improvement protocol was highly effective for producing a more coordinated, efficient, and timely process for administration of antibiotics to patients with open fractures at our hospital. This protocol may be adopted and implemented at other facilities. Therapeutic level III.


Drug use careers and blood-borne pathogen risk behavior in male and female Tanzanian heroin injectors.
Injection drug use in sub-Saharan Africa is a relatively new phenomenon that expands the repertoire of human immunodeficiency virus (HIV)-associated risk behaviors in Africa. We carried out a study of 537 injection drug users (56% men and 44% women) in Dar es Salaam, Tanzania, to examine their HIV risk behaviors and their drug-using careers that had culminated in injecting heroin. Data were collected in 2005-2006 using the Swahili version of the Tanzanian AIDS Prevention Project questionnaire. Marijuana, alcohol, and heroin were the first drugs reported for both men and women. Most drug milestones appeared in a similar order for men and women. Mandrax, khat, and injecting appeared close to one another in chronological time for both men and women, suggesting they were introduced into the country and appeared on the drug scene at about the same (real) time. Drug careers for women were shorter than for men, and time from first use of heroin to first injection was shorter for women. Years of injecting suggested that injecting had increased in males approximately five years prior to data collection, with males injecting earlier, but females being increasingly introduced to injecting in the previous two years. Injecting appears at a mean of five years (men) and three years (women) into their heroin-using career. Heroin use appears to occur in binges, with women being more likely to have sex during a binge. In this sample, more than 90% of women but only 2% of men reported ever trading sex for money. More than 90% of men and women reported using new needles for injection. These data confirm that heroin injecting is well established in large cities in east Africa, and that HIV prevention in the region must now include drug injectors and other drug users.


Health-care costs of losartan and candesartan in the primary treatment of hypertension.
A recent study of two widely used angiotensin receptor blockers reported a reduced risk of cardiovascular events (-14.4%) when using candesartan compared with losartan in the primary treatment of hypertension. In addition to clinical benefits, costs associated with treatment strategies must be considered when allocating scarce health-care resources. The aim of this study was to assess resource use and costs of losartan and candesartan in hypertensive patients. Resource use (drugs, outpatient contacts, hospitalizations and laboratory tests) associated with losartan and candesartan treatment was estimated in 14,100 patients in a real-life clinical setting. We electronically extracted patient data from primary care records and mandatory Swedish national registers for death and hospitalization. Patients treated with losartan had more outpatient contacts (+15.6%), laboratory tests (+13.8%) and hospitalizations (+13.8%) compared with the candesartan group. During a maximum observation time of 9 years, the mean total costs per patient were 10,369 Swedish kronor (95% confidence interval: 3109-17,629) higher in the losartan group. In conclusion, prescribing candesartan for the primary treatment of hypertension results in lower long-term health-care costs compared with losartan.


The phylogeny of the four pan-American MtDNA haplogroups: implications for evolutionary and disease studies.
Only a limited number of complete mitochondrial genome sequences belonging to Native American haplogroups were available until recently, which left America as the continent with the least amount of information about sequence variation of entire mitochondrial DNAs. In this study, a comprehensive overview of all available complete mitochondrial DNA (mtDNA) genomes of the four pan-American haplogroups A2, B2, C1, and D1 is provided by revising the information scattered throughout GenBank and the literature, and adding 14 novel mtDNA sequences. The phylogenies of haplogroups A2, B2, C1, and D1 reveal a large number of sub-haplogroups but suggest that the ancestral Beringian population(s) contributed only six (successful) founder haplotypes to these haplogroups. The derived clades are overall starlike with coalescence times ranging from 18,000 to 21,000 years (with one exception) using the conventional calibration. The average of about 19,000 years somewhat contrasts with the corresponding lower age of about 13,500 years that was recently proposed by employing a different calibration and estimation approach. Our estimate indicates a human entry and spread of the pan-American haplogroups into the Americas right after the peak of the Last Glacial Maximum and comfortably agrees with the undisputed ages of the earliest Paleoindians in South America. In addition, the phylogenetic approach also indicates that the pathogenic status proposed for various mtDNA mutations, which actually define branches of Native American haplogroups, was based on insufficient grounds.


Logical Reduction of Biological Networks to Their Most Determinative Components.
Boolean networks have been widely used as models for gene regulatory networks, signal transduction networks, or neural networks, among many others. One of the main difficulties in analyzing the dynamics of a Boolean network and its sensitivity to perturbations or mutations is the fact that it grows exponentially with the number of nodes. Therefore, various approaches for simplifying the computations and reducing the network to a subset of relevant nodes have been proposed in the past few years. We consider a recently introduced method for reducing a Boolean network to its most determinative nodes that yield the highest information gain. The determinative power of a node is obtained by a summation of all mutual information quantities over all nodes having the chosen node as a common input, thus representing a measure of information gain obtained by the knowledge of the node under consideration. The determinative power of nodes has been considered in the literature under the assumption that the inputs are independent in which case one can use the Bahadur orthonormal basis. In this article, we relax that assumption and use a standard orthonormal basis instead. We use techniques of Hilbert space operators and harmonic analysis to generate formulas for the sensitivity to perturbations of nodes, quantified by the notions of influence, average sensitivity, and strength. Since we work on finite-dimensional spaces, our formulas and estimates can be and are formulated in plain matrix algebra terminology. We analyze the determinative power of nodes for a Boolean model of a signal transduction network of a generic fibroblast cell. We also show the similarities and differences induced by the alternative complete orthonormal basis used. Among the similarities, we mention the fact that the knowledge of the states of the most determinative nodes reduces the entropy or uncertainty of the overall network significantly. In a special case, we obtain a stronger result than in previous works, showing that a large information gain from a set of input nodes generates increased sensitivity to perturbations of those inputs.


Declining mortality in American crow (Corvus brachyrhynchos) following natural West Nile virus infection.
The American crow (Corvus brachyrhynchos) is known to suffer 100% mortality from infection with the New York 1999 strain of West Nile virus (WNV). Following the initial detection of WNV in North America in 1999, we measured prevalence of WNV-reactive antibodies ("seroprevalence") in free-ranging American and fish crows (Corvus ossifragus) of central New Jersey after each transmission season through 2005. In 2002, seroprevalence in American crow juveniles increased to 14% from the 5% of the previous year, potentially indicating increased survival in this species. Using the annual seroprevalence measurements and the number of human West Nile neuroinvasive disease cases as a surrogate for WNV transmission intensity, we developed a model to estimate the annual WNV-associated mortality rates among both of these crow species. Our model supports the hypothesis that mortality is changing over time; the WNV-associated mortality rate declined over time by 1.5% for American crow and by 1.1% for fish crow. The probability that the trend in mortality was negative was 90% for the American crow and 60% for the fish crow.


Insulin Resistance Is Associated With Reduced Mitochondrial Oxidative Capacity Measured by 31P-Magnetic Resonance Spectroscopy in Participants Without Diabetes From the Baltimore Longitudinal Study of Aging.
Whether individuals with insulin resistance (IR) but without criteria for diabetes exhibit reduced mitochondrial oxidative capacity is unclear; addressing this question could guide research for new therapeutics. We investigated 248 participants without diabetes from the Baltimore Longitudinal Study of Aging (BLSA) to determine whether impaired mitochondrial capacity is associated with prediabetes, IR, and duration and severity of hyperglycemia exposure. Mitochondrial capacity was assessed as the postexercise phosphocreatine recovery time constant (τPCr) by 31P-magnetic resonance spectroscopy, with higher τPCr values reflecting reduced capacity. Prediabetes was defined using the American Diabetes Association criteria from fasting and 2-h glucose measurements. IR and sensitivity were calculated using HOMA-IR and Matsuda indices. The duration and severity of hyperglycemia exposure were estimated as the number of years from prediabetes onset and the average oral glucose tolerance test (OGTT) 2-h glucose measurement over previous BLSA visits. Covariates included age, sex, body composition, physical activity, and other confounders. Higher likelihood of prediabetes, higher HOMA-IR, and lower Matsuda index were associated with longer τPCr. Among 205 participants with previous OGTT data, greater severity and longer duration of hyperglycemia were independently associated with longer τPC In conclusion, in individuals without diabetes a more impaired mitochondrial capacity is associated with greater IR and a higher likelihood of prediabetes.


The isolated occlusion of the angular gyri artery. A correlative neurological and anatomical study--case report.
We examined a patient who had signs of a cerebral hemisphere lesion: right hemiparesis, facial weakness, right hemianopsy, acustico-mnestic dysphasia, "empty speech," acalculia, visuo-spatial agnosia and constructional apraxia, but without changes in consciousness. Taking into account clinical signs, computed tomography and carotid angiography findings, we concluded that our patient had an infarction zone in the left temporo-parieto-occipital region, as a consequence of the isolated angular gyri artery (ANG) occlusion. Some clinical signs were a direct effect of the ANG's occlusion. Namely, this artery supplies the cortical regions of great functional significance: the planum polare and temporale, the transverse temporal gyri, the superior and middle temporal gyri, the angular and supramarginal gyri, as well as the superior, middle and inferior occipital gyri. But the other symptoms and signs could be explained by the pathophysiological effect of the cerebral edema on regions supplied by the non-occluded branches of the middle cerebral artery.


The C. elegans LIM homeobox gene lin-11 specifies multiple cell fates during vulval development.
LIM homeobox family members regulate a variety of cell fate choices during animal development. In C. elegans, mutations in the LIM homeobox gene lin-11 have previously been shown to alter the cell division pattern of a subset of the 2 degrees lineage vulval cells. We demonstrate multiple functions of lin-11 during vulval development. We examined the fate of vulval cells in lin-11 mutant animals using five cellular markers and found that lin-11 is necessary for the patterning of both 1 degrees and 2 degrees lineage cells. In the absence of lin-11 function, vulval cells fail to acquire correct identity and inappropriately fuse with each other. The expression pattern of lin-11 reveals dynamic changes during development. Using a temporally controlled overexpression system, we show that lin-11 is initially required in vulval cells for establishing the correct invagination pattern. This process involves asymmetric expression of lin-11 in the 2 degrees lineage cells. Using a conditional RNAi approach, we show that lin-11 regulates vulval morphogenesis. Finally, we show that LDB-1, a NLI/Ldb1/CLIM2 family member, interacts physically with LIN-11, and is necessary for vulval morphogenesis. Together, these findings demonstrate that temporal regulation of lin-11 is crucial for the wild-type vulval patterning.


Working with the team: strategies employed by hospital cancer nurse specialists to implement their role.
This paper describes the strategies used by cancer nurse specialists in the UK to implement their role within the multiprofessional team. The prevalence of cancer increasingly preoccupies the developed world causing concern about the effective use of healthcare resources. The demand to contain healthcare costs and meet the complex needs of patients has led to the development of new and different types of nurse specialist roles. In the UK, these initiatives have been supplemented by reorganizing cancer services to ensure care is delivered by collaborative multiprofessional teams: nurse specialists are considered core team members. While role ambiguity and conflict are acknowledged as barriers to the successful introduction of new roles, little is known about the strategies used by individuals to facilitate role implementation. A grounded theory design using purposive and theoretical sampling. Twenty-nine cancer nurse specialists from five hospitals participated in observation and semistructured interviews. The data were analysed concurrently using the constant comparative method. Acceptance, especially by doctors, was the main problem facing cancer nurse specialists. In addition, they experienced insufficient organizational support for their role. Difficulties with acceptance impaired nurses' ability to provide supportive care to cancer patients. Nurse specialists responded by employing several strategies including building relationships and establishing role boundaries. Some strategies used by nurse specialists are more successful than others in facilitating role implementation. While recommendations exist to assist the introduction of new roles in practice, their implementation by healthcare organizations may be limited. Future approaches should focus on helping nurses develop awareness of the problems they face, why they arise and effective mechanisms for their resolution. The findings highlight the mismatch between cancer policy aspirations and reality and the actions taken by nurse specialists to overcome the problems they encounter.


Factors influencing homelessness in women.
The specific aims of the article were to: (i) compare childhood experiences of intimacy and autonomy as they occurred in the families of origin of women who have and women who have not experienced homelessness; (ii) compare social support, reciprocity, and conflict as they occurred in the childhood support networks and in the current support networks of women who have and women who have not experienced homelessness; (iii) examine the relationships between intimacy and autonomy in families of origin and the social support networks from childhood of women who have and women who have not experienced homelessness. Descriptive correlational design: 255 women were interviewed to determine levels of intimacy, autonomy, social support, reciprocity, and conflict in childhood relationships. The ANCOVA models for each of support, reciprocity, and conflict indicated a significant group effect. The post-hoc analysis for support indicated that the homeless group was significantly lower in support and reciprocity and significantly higher in conflict than the never-homeless groups. The never-homeless, never-abused group scored significantly higher on autonomy and intimacy than the homeless or the never-homeless, abused groups. This study demonstrates the significance of families of origin and learning how to develop and utilize support systems in preventing or reducing homelessness.


BCOR as a novel fusion partner of retinoic acid receptor alpha in a t(X;17)(p11;q12) variant of acute promyelocytic leukemia.
The majority of acute promyelocytic leukemia (APL) cases are characterized by the presence of a promyelocytic leukemia-retinoic acid receptor alpha(RARA) fusion gene. In a small subset, RARA is fused to a different partner, usually involved in regulating cell growth and differentiation. Here, we identified a novel RARA fusion transcript, BCOR-RARA, in a t(X;17)(p11;q12) variant of APL with unique morphologic features, including rectangular and round cytoplasmic inclusion bodies. Although the patient was clinically responsive to all-trans retinoic acid, several relapses occurred with standard chemotherapy and all-trans retinoic acid. BCOR is a transcriptional corepressor through the proto-oncoprotein, BCL6, recruiting histone deacetylases and polycomb repressive complex 1 components. BCOR-RARA was found to possess common features with other RARA fusion proteins. These included: (1) the same break point in RARA cDNA; (2) self-association; (3) retinoid X receptor alpha is necessary for BCOR-RARA to associate with the RARA responsive element; (4) action in a dominant-negative manner on RARA transcriptional activation; and (5) aberrant subcellular relocalization. It should be noted that there was no intact BCOR found in the 45,-Y,t(X;17)(p11;q12) APL cells because they featured only a rearranged X chromosome. These results highlight essential features of pathogenesis in APL in more detail. BCOR appears to be involved not only in human congenital diseases, but also in a human cancer.


Oligomerization of serotonin transporter and its functional consequences.
Two forms of serotonin transporter (SERT) were prepared with different epitope tags. When co-expressed in HeLa cells, the form containing a FLAG tag (Res-FLAG) was associated with the form containing a c-myc tag (Sens-myc). Antibody against c-myc precipitated Res-FLAG from detergent extracts of cells expressing both forms, but not when Res-FLAG was expressed alone. The specificity of the interaction was demonstrated by the observation that anti-myc antibodies did not precipitate the unrelated vesicular stomatitis virus coat glycoprotein when it was co-expressed with Sens-myc. Sens-myc contained a reactive cysteine at position 172, which reacted with both (2-aminoethyl)methanethiosulfonate and N-biotinylaminoethyl methanethiosulfonate on the surface of intact cells. Sens-myc, but not Res-FLAG, was inactivated by these reagents. When co-expressed with Sens-myc, functionally active Res-FLAG was precipitated by immobilized streptavidin from digitonin-solubilized cells that had been treated with N-biotinylaminoethyl methanethiosulfonate. In cells co-expressing mixtures of Sens-myc and Res-FLAG, the amount of inactivation by (2-aminoethyl)methanethiosulfonate was less than expected if the two forms were independent. The results are consistent with a dimeric form of SERT with functional interactions between subunits, and with association of dimers into a higher order complex, possibly a tetramer.


Epistatic and cytonuclear interactions govern outbreeding depression in the autotetraploid Campanulastrum americanum.
The consequences of combining divergent genomes among populations of a diploid species often involve F1 hybrid vigor followed by hybrid breakdown in later recombinant generations. As many as 70% of plant species are thought to have polyploid origins; yet little is known about the genetic architecture of divergence in polyploids and how it may differ from diploid species. We investigated the genetic architecture of population divergence using controlled crosses among five populations of the autotetraploid herb, Campanulastrum americanum. Plants were reciprocally hybridized to produce F1, F2, and F1-backcross generations that were grown with parental types in a greenhouse and measured for performance. In contrast to diploid expectations, most F1 hybrids lacked heterosis and instead showed strong outbreeding depression for early life traits. Recombinant hybrid generations often showed a recovery of performance to levels approximating, or at times even exceeding, the parental values. This pattern was also evident for an index of cumulative fitness. Analyses of line means indicated nonadditive gene action, especially forms of digenic epistasis, often influenced hybrid performance. However, standard diploid genetic models were not adequate for describing the underlying genetic architecture in a number of cases. Differences between reciprocal hybrids indicated that cytoplasmic and/or cytonuclear interactions also contributed to divergence. An enhanced role of epistasis in population differentiation may be the norm in polyploids, which have more gene copies. This study, the first of its kind on a natural autotetraploid, suggests that gene duplication may cause polyploid populations to diverge in a fundamentally different way than diploids.


Practice of gastroenterologists in treating flaring inflammatory bowel disease patients with clostridium difficile: antibiotics alone or combined antibiotics/immunomodulators?
The optimal management of Clostridium difficile infection (CDI) in flaring inflammatory bowel disease (IBD) patients has not been defined. Limited data suggest that coadministration of immunomodulators (IM) with antibiotics (AB) results in a worse outcome. We investigated the prevalent practice among North American gastroenterologists in this scenario. A structured questionnaire presented the clinical cases of two hospitalized patients with ulcerative colitis and concomitant CDI, either with or without prior IM treatment. The questionnaire was distributed to a sample of gastroenterologists at medical centers across North America. Respondents were requested to denote their therapeutic choices for these patients. The survey included 169 gastroenterologists, 122 from the US and 47 from Canada, with an average of 12 ± 10 years of experience in gastroenterology. Forty-two (25%) of the respondents were IBD experts. Seventy-seven (46%) respondents elected to add an IM in combination with AB, whereas 82/169 (54%) treated the flare with AB alone (P = NS). The rate of administering combined AB+IM was similar for the IBD experts and the non-IBD experts. Only 11% of respondents withdrew maintenance azathioprine upon the diagnosis of CDI. More IBD experts stopped azathioprine treatment compared to the non-IBD experts (12/42 versus 6/127, P < 0.001). Overall, 65% of surveyed gastroenterologists stated they believe these patients are afflicted by two simultaneous but separate disease processes. There is significant disagreement among gastroenterologists on whether combination AB+IM or AB alone should be given to IBD patients with CDI-associated flares. Controlled trials are needed to investigate the optimal management approach to this clinical dilemma.


Effect of clearance of bacteria from the blood on the development of systemic bacteraemia in mice.
Clearance of various bacteria isolated from portal and systemic blood of mice was evaluated and compared. All portal blood strains, including Escherichia coli and enterococci were eliminated more rapidly from the circulation than were strains isolated from systemic blood, including Pseudomonas aeruginosa. With mannose-type lectin, mannose or fucose residues that mediated lectinophagocytosis were detected on the surfaces of most portal strains by agglutination tests. Blood clearance of Esch. coli H21 was inhibited by prior injection of mannose into mice, suggesting that the clearance of this strain was mediated by mannose-type lectin on the surface of tissue macrophages. However, no inhibition of clearance of any other strains was observed by the injection of mannose, galactose, or fucose into mice, nor by pre-incubation of bacteria with mannose. Blood clearance of some portal strains was significantly faster in CBA/J mice than in CBA/N mice with B cell immune deficiency, indicating that immunoglobulin was involved in their clearance. Among portal strains only enterococci showed high cell-surface hydrophobicity. These data suggest that initial bacteria blood clearance may be critical in determining whether latent portal bacteraemia progresses to systemic bacteraemia and that the rapid clearance of most strains is multifactorial.


Non-typeable Haemophilus influenzae, an under-recognised pathogen.
Non-typeable Haemophilus influenzae (NTHi) is a major cause of mucosal infections such as otitis media, sinusitis, conjunctivitis, and exacerbations of chronic obstructive pulmonary disease. In some regions, a strong causal relation links this pathogen with infections of the lower respiratory tract. In the past 20 years, a steady but constant increase has occurred in invasive NTHi worldwide, with perinatal infants, young children, and elderly people most at risk. Individuals with underlying comorbidities are most susceptible and infection is associated with high mortality. β-lactamase production is the predominant mechanism of resistance. However, the emergence and spread of β-lactamase-negative ampicillin-resistant strains in many regions of the world is of substantial concern, potentially necessitating changes to antibiotic treatment guidelines for community-acquired infections of the upper and lower respiratory tract and potentially increasing morbidity associated with invasive NTHi infections. Standardised surveillance protocols and typing methodologies to monitor this emerging pathogen should be implemented. International scientific organisations need to raise the profile of NTHi and to document the pathobiology of this microbe.


Optical genome mapping, a promising alternative to gold standard cytogenetic approaches in a series of acute lymphoblastic leukemias.
Acute lymphoblastic leukemias (ALL) are characterized by a large number of cytogenetic abnormalities of clinical interest that require the use of several complementary techniques. Optical genome mapping (OGM) is based on analysis of ultra-high molecular weight DNA molecules that provides a high-resolution genome-wide analysis highlighting copy number and structural anomalies, including balanced translocations. We compared OGM to standard techniques (karyotyping, fluorescent in situ hybridization, single nucleotide polymorphism-array and reverse transcription multiplex ligation-dependent probe amplification) in 10 selected B or T-ALL. Eighty abnormalities were found using standard techniques of which 72 (90%) were correctly detected using OGM. Eight discrepancies were identified, while 12 additional anomalies were found by OGM. Among the discrepancies, four were detected in raw data but not retained because of filtering issues. However, four were truly missed, either because of a low variant allele frequency or because of a low coverage of some regions. Of the additional anomalies revealed by OGM, seven were confirmed by another technique, some of which are recurrent in ALL such as LMO2-TRA and MYC-TRB fusions. Despite false positive anomalies due to background noise and a case of inter-sample contamination secondarily identified, the OGM technology was relatively simple to use with little practice. Thus, OGM represents a promising alternative to cytogenetic techniques currently performed for ALL characterization. It enables a time and cost effective analysis allowing identification of complex cytogenetic events, including those currently inaccessible to standard techniques.


Image processing for cameras with fiber bundle image relay.
Some high-performance imaging systems generate a curved focal surface and so are incompatible with focal plane arrays fabricated by conventional silicon processing. One example is a monocentric lens, which forms a wide field-of-view high-resolution spherical image with a radius equal to the focal length. Optical fiber bundles have been used to couple between this focal surface and planar image sensors. However, such fiber-coupled imaging systems suffer from artifacts due to image sampling and incoherent light transfer by the fiber bundle as well as resampling by the focal plane, resulting in a fixed obscuration pattern. Here, we describe digital image processing techniques to improve image quality in a compact 126° field-of-view, 30 megapixel panoramic imager, where a 12 mm focal length F/1.35 lens made of concentric glass surfaces forms a spherical image surface, which is fiber-coupled to six discrete CMOS focal planes. We characterize the locally space-variant system impulse response at various stages: monocentric lens image formation onto the 2.5 μm pitch fiber bundle, image transfer by the fiber bundle, and sensing by a 1.75 μm pitch backside illuminated color focal plane. We demonstrate methods to mitigate moiré artifacts and local obscuration, correct for sphere to plane mapping distortion and vignetting, and stitch together the image data from discrete sensors into a single panorama. We compare processed images from the prototype to those taken with a 10× larger commercial camera with comparable field-of-view and light collection.


Reduced Lung-Cancer Mortality with Volume CT Screening in a Randomized Trial.
There are limited data from randomized trials regarding whether volume-based, low-dose computed tomographic (CT) screening can reduce lung-cancer mortality among male former and current smokers. A total of 13,195 men (primary analysis) and 2594 women (subgroup analyses) between the ages of 50 and 74 were randomly assigned to undergo CT screening at T0 (baseline), year 1, year 3, and year 5.5 or no screening. We obtained data on cancer diagnosis and the date and cause of death through linkages with national registries in the Netherlands and Belgium, and a review committee confirmed lung cancer as the cause of death when possible. A minimum follow-up of 10 years until December 31, 2015, was completed for all participants. Among men, the average adherence to CT screening was 90.0%. On average, 9.2% of the screened participants underwent at least one additional CT scan (initially indeterminate). The overall referral rate for suspicious nodules was 2.1%. At 10 years of follow-up, the incidence of lung cancer was 5.58 cases per 1000 person-years in the screening group and 4.91 cases per 1000 person-years in the control group; lung-cancer mortality was 2.50 deaths per 1000 person-years and 3.30 deaths per 1000 person-years, respectively. The cumulative rate ratio for death from lung cancer at 10 years was 0.76 (95% confidence interval [CI], 0.61 to 0.94; P = 0.01) in the screening group as compared with the control group, similar to the values at years 8 and 9. Among women, the rate ratio was 0.67 (95% CI, 0.38 to 1.14) at 10 years of follow-up, with values of 0.41 to 0.52 in years 7 through 9. In this trial involving high-risk persons, lung-cancer mortality was significantly lower among those who underwent volume CT screening than among those who underwent no screening. There were low rates of follow-up procedures for results suggestive of lung cancer. (Funded by the Netherlands Organization of Health Research and Development and others; NELSON Netherlands Trial Register number, NL580.).


Nuclear genes affecting albinism in wheat (Triticum aestivum L.) anther culture.
Inheritance of the ability to respond in wheat anther culture was studied from 6×2 reciprocal crosses between six varieties with high and two varieties with low capacity for green plant formation and their parents, replicated in two environments. Effects of genotypes dominated embryo formation and percentages of green plants, accounting for 78.4% and 85.4% of total variation, respectively, while smaller genetic effects were indicated for regeneration. Nuclear genes could explain almost all the genotype effects in this material. Embryo formation showed heterosis over high parent for 5 of the 12 hybrids, while percentages of green plants from the hybrids were intermediate to the parents. General Combining Ability (GCA) could explain 78.8% of the variation for embryo formation among the hybrids, whereas differences in percentage of green plants were dominated by Specific Combining Ability (SCA), accounting for 67.9% of hybrid variation. A positive correlation (r=0.81(**)) was observed between the genetic capacity for regeneration and green plant formation. Analysis of covariance indicated that effects causing GCA for green plant formation were mainly responsible for this correlation. A regression model with two parallel lines divided the six parent lines with high green plant formation into three groups with respect to their reactions with the two testers. The results are discussed with regard to possible involvement of two sets of nuclear genes affecting the percentage of green plants obtained in wheat anther culture: one set consisting of mainly additive effects affecting green plant percentage through an initial effect on regeneration ability, and another set of two or a few more major genes with dominance or epistatic effects uncorrelated with regeneration.


Controlled intoxication: the self-monitoring of excessive alcohol use within a New Zealand tertiary student sample.
Drawn from a study aimed at exploring students' drinking behaviour and attitudes, this article focuses upon findings that revealed how heavy-drinking students monitored and managed their experiences of alcohol intoxication. 819 students residing within three university student residences were invited to participate in three phases of data collection. Utilising a combination of qualitative and quantitative research methods, a total of 15 focus group interviews and 18 indepth interviews were undertaken, and 501 students (61%) completed a written survey questionnaire. Sixty percent of students agreed with the statement "I usually know beforehand if I am going to get drunk". One-half of male drinkers and one-third of female drinkers reported they were intoxicated on a weekly basis. When drinking to intoxication, the majority of students monitored a range of drinking effects (a total of 14 were identified) which they considered were signals for the need to either slow down or stop drinking. The majority of drinkers in this study who consume alcohol with the intention of getting intoxicated, typically drink to a predetermined level of intoxication, and maintain that level by monitoring a range of drinking effects--this behaviour has been termed controlled intoxication. Future harm-minimisation strategies could be developed that encourage heavy-drinkers to adopt 'safer' drinking-effect signals as indicators to slow down or stop drinking.


Autocorrelation evaluation from clipped photon detection.
We present two new techniques for estimating the autocorrelation function, based on the measurement of the mean number of clipped photocounts and on the calculation of the second-order factorial moment of a series of clipped data. The first procedure has the advantage of simplicity, although it produces a poor signal-to-noise ratio. On the other hand, the second technique gives a very good signal-to-noise ratio, when compared with other known methods.


Secondary wastewater disposal for crop irrigation with minimal risks.
A critical objective for any wastewater reuse program is to close the gap between supply of and demand for water and to minimize health and environmental hazards. Thus, the effects of treated effluent on crops, soils and community health must be considered carefully. When applying wastewater to soil-plant systems, it is to be noted that the passage of water through the soil reduces considerably the number of microorganisms carried out by the reclaimed wastewater. Nevertheless, there is a need to study the real rate of organism decay subject to water quality, soil and vegetable characteristics, and irrigation method. The aim of this work is to determine the fate of the fecal coliforms, coliphages F+ and CN13, and helminth eggs survival during the application of reclaimed wastewater in a vineyard orchard near the City of Arad (Israel) via onsurface and subsurface drip irrigation systems. Wastewater obtained from a stabilization pond, and soil samples were tested and an important decrease of microorganisms was reached in both cases, with the better values obtained with the sub-surface drip irrigation system.


Unifying the essential concepts of biological networks: biological insights and philosophical foundations.
Over the last decades, network-based approaches have become highly popular in diverse fields of biology, including neuroscience, ecology, molecular biology and genetics. While these approaches continue to grow very rapidly, some of their conceptual and methodological aspects still require a programmatic foundation. This challenge particularly concerns the question of whether a generalized account of explanatory, organizational and descriptive levels of networks can be applied universally across biological sciences. To this end, this highly interdisciplinary theme issue focuses on the definition, motivation and application of key concepts in biological network science, such as explanatory power of distinctively network explanations, network levels and network hierarchies. This article is part of the theme issue 'Unifying the essential concepts of biological networks: biological insights and philosophical foundations'.


A trauma resource allocation model for ambulances and hospitals.
To develop a mathematical model for the location of trauma care resources. Severely injured patients queried from Maryland hospital discharge and vital statistics data. A spatial injury profile was created by parsing these patients into ZIP codes. The Trauma Resource Allocation Model for Ambulances and Hospitals (TRAMAH) was formulated using integer and heuristic programming. To maximize coverage of severely injured patients, trauma centers and aeromedical depots were simultaneously sited using TRAMAH. A severe injury was considered covered if at least one trauma center was sited within a time standard by ground, or if an aeromedical depot-trauma center pair was sited in such a way that the sum of the flying time from the aeromedical depot to the scene of injury plus the flying time from the scene of injury to the trauma center was within the same time standard. From 1992 to 1994, 26,774 severe injuries were considered for coverage. Across Maryland, 94.8 percent of severely injured residents had access to trauma system resources within 30 minutes and 70.3 percent had access within 15 minutes. For the same number of resources as the existing Maryland Trauma System, TRAMAH achieved a coverage objective of 99.97 percent within 30 minutes. This translated into an additional 461 severely injured people covered each year. Holding in place the trauma centers of the existing system, approximately the same percentage of coverage as that of the existing system was achieved within 15 minutes by optimally locating six fewer aeromedical depots. TRAMAH will allow trauma systems planners to better locate their resources with respect to spatial needs and response times.


Two Loci Contribute Epistastically to Heterospecific Pollen Rejection, a Postmating Isolating Barrier Between Species.
Recognition and rejection of heterospecific male gametes occurs in a broad range of taxa, although the complexity of mechanisms underlying these components of postmating cryptic female choice is poorly understood. In plants, the arena for postmating interactions is the female reproductive tract (pistil), within which heterospecific pollen tube growth can be arrested via active molecular recognition and rejection. Unilateral incompatibility (UI) is one such postmating barrier in which pollen arrest occurs in only one direction of an interspecific cross. We investigated the genetic basis of pistil-side UI between Solanum species, with the specific goal of understanding the role and magnitude of epistasis between UI QTL. Using heterospecific introgression lines (ILs) between Solanum pennellii and S. lycopersicum, we assessed the individual and pairwise effects of three chromosomal regions (ui1.1, ui3.1, and ui12.1) previously associated with interspecific UI among Solanum species. Specifically, we generated double introgression ('pyramided') genotypes that combined ui12.1 with each of ui1.1 and ui3.1, and assessed the strength of UI pollen rejection in the pyramided lines, compared to single introgression genotypes. We found that none of the three QTL individually showed UI rejection phenotypes, but lines combining ui3.1 and ui12.1 showed significant pistil-side pollen rejection. Furthermore, double ILs (DILs) that combined different chromosomal regions overlapping ui3.1 differed significantly in their rate of UI, consistent with at least two genetic factors on chromosome three contributing quantitatively to interspecific pollen rejection. Together, our data indicate that loci on both chromosomes 3 and 12 are jointly required for the expression of UI between S. pennellii and S. lycopersicum, suggesting that coordinated molecular interactions among a relatively few loci underlie the expression of this postmating prezygotic barrier. In addition, in conjunction with previous data, at least one of these loci appears to also contribute to conspecific self-incompatibility (SI), consistent with a partially shared genetic basis between inter- and intraspecific mechanisms of postmating prezygotic female choice.


Causes and methods to estimate cryptic sources of fishing mortality.
Cryptic, not readily detectable, components of fishing mortality are not routinely accounted for in fisheries management because of a lack of adequate data, and for some components, a lack of accurate estimation methods. Cryptic fishing mortalities can cause adverse ecological effects, are a source of wastage, reduce the sustainability of fishery resources and, when unaccounted for, can cause errors in stock assessments and population models. Sources of cryptic fishing mortality are (1) pre-catch losses, where catch dies from the fishing operation but is not brought onboard when the gear is retrieved, (2) ghost-fishing mortality by fishing gear that was abandoned, lost or discarded, (3) post-release mortality of catch that is retrieved and then released alive but later dies as a result of stress and injury sustained from the fishing interaction, (4) collateral mortalities indirectly caused by various ecological effects of fishing and (5) losses due to synergistic effects of multiple interacting sources of stress and injury from fishing operations, or from cumulative stress and injury caused by repeated sub-lethal interactions with fishing operations. To fill a gap in international guidance on best practices, causes and methods for estimating each component of cryptic fishing mortality are described, and considerations for their effective application are identified. Research priorities to fill gaps in understanding the causes and estimating cryptic mortality are highlighted.


The Effects of Hyperbaric Oxygen at Different Pressures on Oxidative Stress and Antioxidant Status in Rats.
Background: The optimal use of oxygen at greater than atmospheric pressures in any operational or therapeutic application (hyperbaric oxygen, HBO2) requires awareness of the fact that the beneficial effects of oxygen coexist with toxic effects depending on the pressure and duration of exposure. In this study, we aimed to investigate the effect of HBO2 therapy on oxidative stress and antioxidant status in commonly used protocol for acute HBO2 indications, such as carbon monoxide intoxication, central retinal artery occlusion, crush injury, gas gangrene, and to compare it with normobaric oxygen (NBO2) in healthy rats. Materials and Methods: Fifty-six male, young adult Wistar albino rats were randomly divided into seven groups and named as Group I through Group VII. Plasma malondialdehyde (MDA), superoxide dismutase (SOD), and erythrocyte glutathione (GSH) levels in control group were compared to the levels in other groups. Results: The increases in MDA levels and the decrease in SOD activities were statistically significant in HBO2 groups at the end of the first 24 h when compared to the control group, and the significant decrease in erythrocyte GSH level was only at 2.4 atmospheres absolute. Conclusions: The present study showed that pressure and frequency of exposure are important factors to consider when investigating HBO2-induced oxidative stress and antioxidant response.


Effect of ganglioside GT1b on the in vitro maturation of porcine oocytes and embryonic development.
Ganglioside is an acidic glycosphingolipid with sialic acids residues. This study was performed to investigate the effect and mechanism of ganglioside GT1b in porcine oocytes in the process of in vitro maturation (IVM) and preimplantation development. Metaphase II (MII) rates were significantly (P < 0.05) different between the control group and the 5 nM GT1b treatment group. Intracellular glutathione (GSH) levels in oocytes matured with 5 nM and 20 nM and GT1b decreased significantly (P < 0.05). The 10 nM group showed a significant (P < 0.05) decrease in intracellular reactive oxygen species (ROS) levels compared with the control group. Subsequently, the level of intracellular Ca(2+) in oocytes treated with different concentrations of GT1b was measured. Intracellular Ca(2+) was significantly (P < 0.05) increased with a higher concentration of GT1b in a dose-dependent manner. Real-time PCR was performed and showed that the expression of bradykinin 2 receptor (B2R) and calcium/calmodulin-dependent protein kinase II delta (CaMKIIδ) in cumulus cells was significantly (P < 0.05) decreased in the 20 nM GT1b treatment group. Treatment with 5 nM GT1b significantly (P < 0.05) decreased the expression of CaMKIIδ. In oocytes, treatment with 5 nM GT1b significantly (P < 0.05) decreased CaMKIIγ and POU5F1 (POU domain, class 5, transcription factor 1). However, treatment with 20 nM GT1b significantly (P < 0.05) increased the expression of POU5F1. Finally, embryonic developmental data showed no significant differences in the two experiments (parthenogenesis and in vitro fertilization). In conclusion, the results of the present study indicated that GT1b plays an important role in increasing the nuclear maturation rate and decreasing the intracellular ROS levels during IVM. However, GT1b inhibited maturation of the cytoplasm by maintaining intracellular Ca(2+) in the process of oocyte maturation regardless of the cell cycle stage. Therefore, GT1b is thought to act on another mechanism that controls intracellular Ca(2+).


Owned and unowned dog population estimation, dog management and dog bites to inform rabies prevention and response on Lombok Island, Indonesia.
Although Indonesia has been rabies-infected since at least the 1880s, some islands remain rabies-free, such as Lombok. However, due to its adjacency to rabies-infected islands such as Bali and Flores, there is considerable risk of a rabies incursion. As part of a rabies risk assessment project, surveys were conducted to estimate the size of the dog population and to describe dog management practices of households belonging to different ethnic groups. A photographic-recapture method was employed and the number of unowned dogs was estimated. A total of 400 dog owning households were interviewed, 300 at an urban site and 100 at a rural site. The majority of the interviewed households belonged to the Balinese ethnic group. Owned dogs were more likely male, and non-pedigree or local breed. These households kept their dogs either fully restricted, semi-free roaming or free-roaming but full restriction was reported only at the urban site. Dog bite cases were reported to be higher at the urban site, and commonly affected children/young adults to 20 years old and males. A higher number of unowned dogs was observed at the urban site than at the rural site. Data generated within these surveys can inform rabies risk assessment models to quantify the probability of rabies being released into Lombok and resulting in the infection of the local dog population. The information gained is critical for efforts to educate dog owners about rabies, as a component of preparedness to prevent the establishment of rabies should an incursion occur.


Angiocrine signals regulate quiescence and therapy resistance in bone metastasis.
Bone provides supportive microenvironments for hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) and is a frequent site of metastasis. While incidences of bone metastases increase with age, the properties of the bone marrow microenvironment that regulate dormancy and reactivation of disseminated tumor cells (DTCs) remain poorly understood. Here, we elucidate the age-associated changes in the bone secretome that trigger proliferation of HSCs, MSCs, and DTCs in the aging bone marrow microenvironment. Remarkably, a bone-specific mechanism involving expansion of pericytes and induction of quiescence-promoting secretome rendered this proliferative microenvironment resistant to radiation and chemotherapy. This bone-specific expansion of pericytes was triggered by an increase in PDGF signaling via remodeling of specialized type H blood vessels in response to therapy. The decline in bone marrow pericytes upon aging provides an explanation for loss of quiescence and expansion of cancer cells in the aged bone marrow microenvironment. Manipulation of blood flow - specifically, reduced blood flow - inhibited pericyte expansion, regulated endothelial PDGF-B expression, and rendered bone metastatic cancer cells susceptible to radiation and chemotherapy. Thus, our study provides a framework to recognize bone marrow vascular niches in age-associated increases in metastasis and to target angiocrine signals in therapeutic strategies to manage bone metastasis.


Antibiotic treatment for Burkholderia cepacia complex in people with cystic fibrosis experiencing a pulmonary exacerbation.
Chronic pulmonary infection is a hallmark of lung disease in cystic fibrosis. Infections dominated by organisms of the Burkholderia cepacia complex, a group of at least 18 closely-related species of gram-negative bacteria, are particularly difficult to treat. These infections may be associated with a fulminant necrotising pneumonia. Burkholderia cepacia complex bacteria are resistant to many common antibiotics and able to acquire resistance against many more. Following patient segregation in cystic fibrosis medical care, the more virulent epidemic strains are not as frequent, and new infections are more likely to be with less virulent environmentally-acquired strains. Although evidence-based guidelines exist for treating respiratory exacerbations involving Pseudomonas aeruginosa, these cannot be extended to Burkholderia cepacia complex infections. This review, which is an update of a previous review, aims to assess the available trial evidence for the choice and application of treatments for these infections. To assess the effectiveness and safety of different antibiotic regimens in people with cystic fibrosis experiencing an exacerbation and chronically infected with organisms of the Burkholderia cepacia complex. We searched the Cochrane Cystic Fibrosis Trials Register, compiled from electronic database searches and handsearching of journals and conference abstract books. We also searched the reference lists of relevant articles and reviews.Date of latest search: 28 August 2015. Randomised and quasi-randomised controlled trials of treatments for exacerbations of pulmonary symptoms in people with cystic fibrosis chronically infected with organisms of the Burkholderia cepacia complex. No relevant trials were identified. No trials were included in this review. Burkholderia cepacia complex infections present a significant challenge for people with cystic fibrosis and their clinicians. The incidence is likely to increase as the cystic fibrosis population ages; and managing and treating these infections will become more important. There is a lack of trial evidence to guide decision making and no conclusions can be drawn from this review about the optimal antibiotic regimens for people with cystic fibrosis who have chronic Burkholderia cepacia complex infections. Clinicians must continue to assess each person individually, taking into account in vitro antibiotic susceptibility data, previous clinical responses and their own experience. Multicentre randomised clinical trials are needed to assess the effectiveness of different antibiotic regimens in people with cystic fibrosis infected with organisms of the Burkholderia cepacia complex.


IFAP 300 is common to desmosomes and hemidesmosomes and is a possible linker of intermediate filaments to these junctions.
The distribution of IFAP 300, a protein previously characterized as cross-linking vimentin intermediate filaments (IF), has been investigated in epithelial cells. In frozen sections of bovine tongue epithelium the staining obtained with IFAP 300 antibodies is concentrated in the peripheral cytoplasm of keratinocytes, including the entire peripheral region of basal cells. Further immunofluorescence studies reveal that in primary cultures of mouse keratinocytes the distribution of IFAP 300 is similar to that of the desmosomal protein desmoplakin. In rat bladder carcinoma 804G cells the staining pattern of IFAP 300 antibodies coincides with that obtained with antibodies against the hemidesmosomal protein BP 230. By immunogold electron microscopy IFAP 300 is mainly located at sites where IF appear to attach to desmosomes and hemidesmosomes. Morphometric analyses of the distribution of the gold particles show that IFAP 300 overlaps with desmoplakin and BP 230, but also that it extends deeper into the cytoplasm than these latter two proteins. The staining reaction seen in epithelial cells by immunofluorescence and immunogold is specific for IFAP 300 as shown by immunoblotting. Immunoblotting also reveals that IFAP 300 is present in both cell-free preparations of desmosomes and hemidesmosomes. These morphological and biochemical results are intriguing since, in recent years, the proteins appearing in these two types of junctions have been found to be different. One possible exception is plectin, a protein that has been suggested to be very similar to IFAP 300. However, we show here that IFAP 300 differs from plectin in several respects, including differences at the primary sequence level. We also show that purified IFAP 300 pellets with in vitro polymerized IF prepared from desmosome-associated keratins under conditions in which IFAP 300 alone is not sedimentable. This indicates that IFAP 300 can associate with keratin IF. These data, taken together with the immunogold results, suggest that IFAP 300 functions in epithelial cells as a linker protein connecting IF to desmosomes as well as to hemidesmosomes, possibly through structurally related proteins such as desmoplakin and BP 230, respectively.


Cannabidiol Inhibition of Murine Primary Nociceptors: Tight Binding to Slow Inactivated States of Nav1.8 Channels.
The nonpsychoactive phytocannabinoid cannabidiol (CBD) has been shown to have analgesic effects in animal studies but little is known about its mechanism of action. We examined the effects of CBD on intrinsic excitability of primary pain-sensing neurons. Studying acutely dissociated capsaicin-sensitive mouse DRG neurons at 37°C, we found that CBD effectively inhibited repetitive action potential firing, from 15-20 action potentials evoked by 1 s current injections in control to 1-3 action potentials with 2 μm CBD. Reduction of repetitive firing was accompanied by a reduction of action potential height, widening of action potentials, reduction of the afterhyperpolarization, and increased propensity to enter depolarization block. Voltage-clamp experiments showed that CBD inhibited both TTX-sensitive and TTX-resistant (TTX-R) sodium currents in a use-dependent manner. CBD showed strong state-dependent inhibition of TTX-R channels, with fast binding to inactivated channels during depolarizations and slow unbinding on repolarization. CBD alteration of channel availability at various voltages suggested that CBD binds especially tightly [Kd (dissociation constant), ∼150 nm] to the slow inactivated state of TTX-R channels, which can be substantially occupied at voltages as negative as -40 mV. Remarkably, CBD was more potent in inhibiting TTX-R channels and inhibiting action potential firing than the local anesthetic bupivacaine. We conclude that CBD might produce some of its analgesic effects by direct effects on neuronal excitability, with tight binding to the slow inactivated state of Nav1.8 channels contributing to effective inhibition of repetitive firing by modest depolarizations.SIGNIFICANCE STATEMENT Cannabidiol (CBD) has been shown to inhibit pain in various rodent models, but the mechanism of this effect is unknown. We describe the ability of CBD to inhibit repetitive action potential firing in primary nociceptive neurons from mouse dorsal root ganglia and analyze the effects on voltage-dependent sodium channels. We find that CBD interacts with TTX-resistant sodium channels in a state-dependent manner suggesting particularly tight binding to slow inactivated states of Nav1.8 channels, which dominate the overall inactivation of Nav1.8 channels for small maintained depolarizations from the resting potential. The results suggest that CBD can exert analgesic effects in part by directly inhibiting repetitive firing of primary nociceptors and suggest a strategy of identifying compounds that bind selectively to slow inactivated states of Nav1.8 channels for developing effective analgesics.


Improved Saunders method for the analysis of lateral shearing interferograms.
An interferogram obtained by use of ordinary interferometers, such as Fizeau and Twyman-Green interferometers, will show a contour map of the wave front under test. A lateral-shearing interferogram, however, will show a contour map of the difference between the wave front under test and a sheared wave front, that is, a contour map of the derivative of the wave front under test. Therefore one can reconstruct the shape of the wave front under test by analyzing that difference. Many methods for reconstructing a wave front have been proposed. The Saunders method reconstructs a wave front; rapidly however the wave-front data are reconstructed only at intervals of the amount of shear along the direction of the shear. Therefore the method has low spatial resolution. A method for reconstructing a wave front that is based on the Saunders method and has high spatial resolution is proposed. The method analyzes the differences that are produced by shearing of the wave front under test in many directions. This method requires a large number of interferograms for reconstructing the wave front. Here the method is described, and its validity is confirmed by simulation.


Linkage of a gene causing high bone mass to human chromosome 11 (11q12-13)
The purpose of this paper is to report the linkage of a genetic locus (designated "HBM") in the human genome to a phenotype of very high spinal bone density, using a single extended pedigree. We measured spinal bone-mineral density, spinal Z(BMD), and collected blood from 22 members of this kindred. DNA was genotyped on an Applied Biosystems model 377 (ABI PRISM Linkage Mapping Sets; Perkin Elmer Applied Biosystems), by use of fluorescence-based marker sets that included 345 markers. Both two-point and multipoint linkage analyses were performed, by use of affected/unaffected and quantitative-trait models. Spinal Z(BMD) for affected individuals (N = 12) of the kindred was 5.54 +/- 1.40; and for unaffected individuals (N = 16) it was 0.41 +/- 0.81. The trait was present in affected individuals 18-86 years of age, suggesting that HBM influences peak bone mass. The only region of linkage was to a series of markers on chromosome 11 (11q12-13). The highest LOD score (5.21) obtained in two-point analysis, when a quantitative-trait model was used, was at D11S987. Multipoint analysis using a quantitative-trait model confirmed the linkage, with a LOD score of 5.74 near marker D11S987. HBM demonstrates the utility of spinal Z(BMD) as a quantitative bone phenotype that can be used for linkage analysis. Osteoporosis pseudoglioma syndrome also has been mapped to this region of chromosome 11. Identification of the causal gene for both traits will be required for determination of whether a single gene with different alleles that determine a wide range of peak bone densities exists in this region.


Artificial cytochrome b: computer modeling and evaluation of redox potentials.
We generated atomic coordinates of an artificial protein that was recently synthesized to model the central part of the native cytochrome b (Cb) subunit consisting of a four-helix bundle with two hemes. Since no X-ray structure is available, the structural elements of the artificial Cb were assembled from scratch using all known chemical and structural information available and avoiding strain as much as possible. Molecular dynamics (MD) simulations applied to this model protein exhibited root-mean-square deviations as small as those obtained from MD simulations starting with the crystal structure of the native Cb subunit. This demonstrates that the modeled structure of the artificial Cb is relatively rigid and strain-free. The model structure of the artificial Cb was used to determine the redox potentials of the two hemes by calculating the electrostatic energies from the solution of the linearized Poisson-Boltzmann equation (LPBE). The calculated redox potentials agree within 20 meV with the experimentally measured values. The dependence of the redox potentials of the hemes on the protein environment was analyzed. Accordingly, the total shift in the redox potentials is mainly due to the low dielectric medium of the protein, the protein backbone charges, and the salt bridges formed between the arginines and the propionic acid groups of the hemes. The difference in the shift of the redox potentials is due to the interactions with the hydrophilic side chains and the salt bridges formed with the propionic acids of the hemes. For comparison and to test the computational procedure, the redox potentials of the two hemes in the native Cb from the cytochrome bc(1) (Cbc(1)) complex were also calculated. Also in this case the computed redox potentials agree well with experiments.


Singlet oxygen photosensitisation by the fluorescent protein Pp2FbFP L30M, a novel derivative of Pseudomonas putida flavin-binding Pp2FbFP.
Flavin-binding fluorescent proteins (FbFPs) are a class of fluorescent reporters that have been increasingly used as reporters in the study of cellular structures and dynamics. Flavin's intrinsic high singlet oxygen ((1)O2) quantum yield (ΦΔ = 0.51) provides a basis for the development of new FbFP mutants capable of photosensitising (1)O2 for mechanistic and therapeutic applications, as recently exemplified by the FbFP miniSOG. In the present work we report an investigation on the (1)O2 photoproduction by Pp2FbFP L30M, a novel derivative of Pseudomonas putida Pp2FbFP. Direct detection of (1)O2 through its phosphorescence at 1275 nm yielded the value ΦΔ = 0.09 ± 0.01, which is the highest (1)O2 quantum yield reported to date for any FP and is approximately 3-fold higher than the ΦΔ for miniSOG. Unlike miniSOG, transient absorption measurements revealed the existence of two independent triplet states each with a different ability to sensitise (1)O2.


Is there different risk of cancer among end-stage renal disease patients undergoing hemodialysis and peritoneal dialysis?
Cancer is a global issue in recent decade. Despite this alarming increase in the incidence of cancer, to date, whether the risk of developing cancer differs among peritoneal dialysis (PD) and hemodialysis (HD) patients is still uncertain. In this retrospective cohort study, data were obtained from the National Health Insurance Research Database of Taiwan, which provides coverage to almost 99% of the nation's population. After matching, a total of 4491 (or 3369) incident PD patients and 8982 (or 6738) incident HD patients between 2000 and 2009 were enrolled from the database. In addition, 22,455 (or 16,845) nondialysis patients were selected as a control group. The patients were monitored for the occurrence of cancer until 2010, and their data were analyzed using several different models. In general, the results showed that the risks of hepatocellular, kidney, bladder, extra kidney/bladder urinary tract, and thyroid cancers were higher in dialysis patients. We also compared the risk of cancer between two dialysis groups by using the HD patients as the reference group. The result showed that there is no significant different for each cancer risk between two dialysis groups. In conclusion, dialysis patients had a higher risk of certain types of cancer than those in the nonuremia group. However, there was no significant difference in the cancer risk between the two dialysis groups when compared directly.


Endovascular occlusion of the carotid or vertebral artery with temporary proximal flow arrest and microcoils: clinical results.
To determine the clinical effectiveness of parent artery occlusion of the carotid or vertebral artery by means of temporary proximal flow arrest and microcoils. Nineteen parent artery occlusions (15 carotid, four vertebral) were performed in 19 patients who successfully passed a balloon test occlusion. In these patients, endovascular occlusion of the carotid or vertebral artery was accomplished with the use of temporary proximal flow arrest and microcoils. All 19 parent arteries were occluded. Eighteen patients (95%) had good outcomes and one (5%) had a poor outcome. Fourteen patients (74%) had no complications and five (26%) had complications, of whom only one was left with a permanent neurologic deficit. Three (60%) of the complications were the result of delayed ischemic events after parent artery occlusion and were not predicted by balloon test occlusion. Endovascular occlusion with temporary proximal flow arrest and microcoils can be done effectively and successfully. The predictive value of the balloon test occlusion is the major complicating factor, as it is with balloon occlusion. This technique offers an additional tool that can be used for endovascular occlusion of the carotid or vertebral artery and seems to be less difficult technically. It is our primary technique for parent artery occlusion.


Surface dynamics of aerolysin on the plasma membrane of living cells.
Aerolysin secreted by the human pathogen Aeromonas hydrophila belongs to a group of bacterial toxins that are hemolytic and form channels in biological membranes. The toxin is secreted as an inactive precursor proaerolysin that must be proteolytically processed at its C-terminus to become active. The toxin then polymerizes into a heptameric ring that is amphipathic and can insert into a lipid bilayer and form a pore. We have examined these various steps at the surface of target cells. The toxin binds to specific receptors. Various receptors have been identified, all of which are anchored to the plasma membrane via a glycosylphosphatidyl inositol (GPI)-anchored moiety. The GPI anchor confers to the protein that is linked to it two usual properties: (i) the protein has a higher lateral mobility in a phospholipid bilayer than its transmembrane counterpart, (ii) the protein has the capacity to transiently associate with cholesterol-glycosphingolipid-rich microdomains. We have shown that both these properties of GPI-anchored proteins are exploited by proaerolysin bound to its receptor. The high lateral mobility within the phosphoglyceride region of the plasma membrane favors the encounter of the protoxin with its converting enzyme furin. The ability to associate with microdomains on the other hand favors the oligomerization process presumably by concentrating the toxin locally.


Joint 2D and 3D phase processing for quantitative susceptibility mapping: application to 2D echo-planar imaging.
Quantitative susceptibility mapping (QSM) measures tissue magnetic susceptibility and typically relies on time-consuming three-dimensional (3D) gradient-echo (GRE) MRI. Recent studies have shown that two-dimensional (2D) multi-slice gradient-echo echo-planar imaging (GRE-EPI), which is commonly used in functional MRI (fMRI) and other dynamic imaging techniques, can also be used to produce data suitable for QSM with much shorter scan times. However, the production of high-quality QSM maps is difficult because data obtained by 2D multi-slice scans often have phase inconsistencies across adjacent slices and strong susceptibility field gradients near air-tissue interfaces. To address these challenges in 2D EPI-based QSM studies, we present a new data processing procedure that integrates 2D and 3D phase processing. First, 2D Laplacian-based phase unwrapping and 2D background phase removal are performed to reduce phase inconsistencies between slices and remove in-plane harmonic components of the background phase. This is followed by 3D background phase removal for the through-plane harmonic components. The proposed phase processing was evaluated with 2D EPI data obtained from healthy volunteers, and compared against conventional 3D phase processing using the same 2D EPI datasets. Our QSM results were also compared with QSM values from time-consuming 3D GRE data, which were taken as ground truth. The experimental results show that this new 2D EPI-based QSM technique can produce quantitative susceptibility measures that are comparable with those of 3D GRE-based QSM across different brain regions (e.g. subcortical iron-rich gray matter, cortical gray and white matter). This new 2D EPI QSM reconstruction method is implemented within STI Suite, which is a comprehensive shareware for susceptibility imaging and quantification. Copyright © 2016 John Wiley & Sons, Ltd.


Radiographic, biomechanical, and histological evaluation of rhBMP-2 in a 3-level intertransverse process spine fusion: an ovine study.
OBJECTIVE The objective of this study was to evaluate bone grafts consisting of rhBMP-2 on an absorbable collagen sponge with a ceramic composite bulking agent, rhBMP-2, directly on a ceramic-collagen sponge carrier or iliac crest bone graft (ICBG) in combination with local bone graft to effect fusion in a multisegmental instrumented ovine lumbar intertransverse process fusion model. METHODS Thirty-six sheep had a single treatment at 3 spinal levels in both the right and left intertransverse process spaces. Group 1 sheep were treated with 7.5 cm3 of autograft consisting of ICBG plus local bone for each intertransverse process space. For Groups 2-4, 4 cm3 of local bone was placed within the intertransverse process space followed by 4.5-5 cm3 of the rhBMP-2 graft material. Group 2 animals received 1.5 mg/cm3 rhBMP-2 on an absorbable collagen sponge with a commercial bone void filler consisting of Type I lyophilized collagen with a biphasic hydroxyapatite/β-tricalcium phosphate ceramic with local bone. Group 3 animals received 0.75 mg/m cm3 of rhBMP-2 on a collagen ceramic sponge carrier with local bone. Group 4 animals received 1.35 mg/cm3 of rhBMP-2 on the same collagen ceramic sponge carrier with local bone. Sheep were euthanized 6 months postoperatively. Manual palpation, biomechanical testing, CT, radiography, and undecalcified histology were performed to assess the presence of fusion associated with the treatments. RESULTS All animals in Groups 2-4 that received grafts containing rhBMP-2 achieved radiographic and CT fusion at all 3 levels. In Group 1 (bone autograft alone), only 19% of the levels demonstrated radiographic fusion, 14% resulted in possible radiographic fusion, and 67% of the levels demonstrated radiographic nonfusion. Biomechanical testing showed that Groups 2-4 demonstrated similar stiffness of the L2-5 segment in all 6 loading directions, with each of the 3 groups having significantly greater stiffness than the autograft-only group. In Group 1, only 2 of 18 levels were rated as achieving bilateral histological fusion, with an additional 3 levels showing a unilateral fusion. The majority of the treated levels (13/18) in Group 1 were scored as histological nonfusions. There were no histological nonfusions in Groups 2 through 4. All 18 levels in Group 2 were rated as bilateral histological fusions. A majority (34/36) of the levels in Group 3 were rated as bilateral histological fusions, with 2 levels showing a unilateral fusion. A majority (35/36) of the levels in Group 4 were rated as bilateral histological fusions, with 1 level showing a unilateral fusion. CONCLUSIONS In the ovine multilevel instrumented intertransverse process fusion model, rhBMP-2 was able to consistently achieve CT, radiographic, biomechanical, and histological fusion. Compared with ICBG, the gold standard for bone grafting, rhBMP-2 was statistically superior at achieving radiographic and histological fusion.


Comparison of short-term results between the modified Karydakis flap and the modified Limberg flap in the management of pilonidal sinus disease: a randomized controlled study.
The modified Karydakis flap and the modified Limberg flap are commonly used in the surgical management of sacrococcygeal pilonidal sinus disease. The study aimed to compare short- term results of these techniques. The modified Karydakis flap and the modified Limberg flap were compared in a randomized controlled trial. The study was conducted in the day-case surgery department of a large academic teaching hospital in Alexandria, Egypt, from February 2009 through September 2011. Patients undergoing surgery for chronic sacrococcygeal pilonidal sinus disease were eligible for the study. Patients with sepsis were eligible only after aggressive treatment to eliminate sepsis. Patients were randomly allocated to undergo surgery with either the modified Karydakis flap or the modified Limberg flap. The primary end point was operative time. Secondary endpoints included proportion of patients with postoperative complications, proportion of patients who were satisfied with the cosmetic results, and the rate of recurrence. Follow-up times were not sufficient to evaluate long-term recurrence rates. Of 154 patients screened, 125 were enrolled, and 120 patients completed the study. The median operative time was significantly shorter in patients with the modified Karydakis flap than in those with the modified Limberg flap: 33 (range, 28-40) min vs 52 (range, 48-62) min; p < 0.001). No significant differences were found between study groups regarding overall complication rate (23% vs 40%, p = 0.08), wound infection (3% vs 5%, p > 0.99), subcutaneous fluid collection (5% vs 0%, p = 0.24), or hypoesthesia (10% vs 23%, p = 0.09). Full-thickness wound disruption was encountered in 9 patients (15%) in the modified Limberg group vs no patient in the modified Karydakis group (p = 0.003). The median follow-up duration was 20.5 months in each study group. One patient (2%) in the modified Karydakis group developed recurrent disease vs 2 patients (3%) in the modified Limberg group (p > 0.99). In the modified Karydakis group, 58 patients (97%) were satisfied with the cosmetic outcome and were willing to recommend the operation to others vs 43 patients (72%) in the modified Limberg group (p < 0.001). Both techniques provide effective treatment for pilonidal sinus disease and can be performed safely as day-case surgery. The modified Karydakis flap is associated with significantly shorter operative time, a lower full-thickness wound disruption rate, and a higher patient satisfaction rate.


ORR viability of alumina-supported platinum nanocluster: exploring oxidation behaviour by DFT.
While alumina-supported platinum particles are versatile for several oxidation reactions, their viability as ORR catalysts has not been explored to date. Therefore, to assess the prospects of alumina-supported platinum nanoclusters in ORRs, a systematic DFT study has been carried out to explore the oxidation behavior of a Ptn@Al2O3 (n = 1-7, 10) cluster. The results are compared with the oxidation behavior of the corresponding gas phase platinum cluster and that of an extended Pt(111) slab. Both supported and unsupported clusters activate adsorbed oxygen molecules and energetically favor dissociative chemisorption of oxygen, leading to stable oxide formation with Pt-O-Pt linkages. However, the influence of the alumina substrate downshifts the d-band centre of the platinum cluster, which not only reduces the reaction enthalpy of oxidation by 8-10%, but also elongates the Pt-O bond of the oxide product by 3-8%. These observations indicate that removal of oxide will be relatively easier for supported clusters than for unsupported clusters. Cluster binding is found to sustain during oxidation, as oxidation of the platinum host cluster results in reduction of the distance between the cluster and support surface. While the gas phase Pt10 cluster does not show any similarity to the oxidation behavior shown by THE Pt(111) slab, the Pt10@Al2O3 cluster reveals close resemblance. Both the Pt(111) slab and Pt10@Al2O3 cluster form similar oxide products, having tri-coordinated oxygen with comparable Pt-O bond distances. The observed resemblance has been attributed to the similarity in the electronic structure and d-band centre position of the platinum surface and alumina-supported Pt10 cluster. Whilst this similar oxidation behaviour of the Pt10@Al2O3 cluster endorses its viability as an ORR catalyst, further modulation of this catalyst is desirable to improve its potential.


Structure and activity of alpha-conotoxin PeIA at nicotinic acetylcholine receptor subtypes and GABA(B) receptor-coupled N-type calcium channels.
α-Conotoxins are peptides from cone snails that target the nicotinic acetylcholine receptor (nAChR). RgIA and Vc1.1 have analgesic activity in animal pain models. Both peptides target the α9α10 nAChR and inhibit N-type calcium channels via GABA(B) receptor activation, but the mechanism of action of analgesic activity is unknown. PeIA has previously been shown to inhibit the α9α10 and α3β2 nAChRs. In this study, we have determined the structure of PeIA and shown that it is also a potent inhibitor of N-type calcium channels via GABA(B) receptor activation. The characteristic α-conotoxin fold is present in PeIA, but it has a different distribution of surface-exposed hydrophobic and charged residues compared with Vc1.1. Thus, the surface residue distribution, rather than the overall fold, appears to be responsible for the 50-fold increase in selectivity at the α3β2 nAChR by PeIA relative to Vc1.1. In contrast to their difference in potency at the nAChR, the equipotent activity of PeIA and Vc1.1 at the GABA(B) receptor suggests that the GABA(B) receptor is more tolerant to changes in surface residues than is the nAChR. The conserved Asp-Pro-Arg motif of Vc1.1 and RgIA, which is crucial for potency at the α9α10 nAChR, is not required for activity at GABA(B) receptor/N-type calcium channels because PeIA has a His-Pro-Ala motif in the equivalent position. This study shows that different structure-activity relationships are associated with the targeting of the GABA(B) receptor versus nAChRs. Furthermore, there is probably a much more diverse range of conotoxins that target the GABA(B) receptor than currently realized.


Cutting edge: Decreased accumulation and regulatory function of CD4+ CD25(high) T cells in human STAT5b deficiency.
We show that STAT5b is important for the in vivo accumulation of CD4+ CD25(high) T cells with regulatory cell function. A patient homozygous for a missense A630P STAT5b mutation displayed immune dysregulation and decreased numbers of CD4+ CD25(high) T cells. STAT5b(A630P/A630P) CD4+ CD25(high) T cells had low expression of forkhead box P3 and an impaired ability to suppress the proliferation of or to kill CD4+ CD25- T cells. Expression of CD25, a component of the high-affinity IL-2R, was also reduced in response to IL-2 or after in vitro propagation. The impact of the STAT5b mutation was selective in that IL-2-mediated up-regulation of the common gamma-chain cytokine receptor and perforin, and activation-induced expressions of CD154 and IFN-gamma were normal. These results indicate that STAT5b propagates an important IL-2-mediated signal for the in vivo accumulation of functional regulatory T cells.


Location of knee pain in medial knee osteoarthritis: patterns and associations with self-reported clinical symptoms.
To (1) document pain location in medial tibiofemoral osteoarthritis (OA) using the patient-administered Photographic Knee Pain Map (PKPM); (2) compare pain severity, nature and likelihood of neuropathic-like symptoms, physical dysfunction and presence of symptoms at other sites across the most common pain patterns. Baseline data were analysed from 164 participants with medial tibiofemoral OA (TFJOA) participating in a randomised controlled trial (RCT). Participants completed the PKPM indicating all relevant pain zones of their most painful knee. Pain zones were collapsed into regions to determine patterns of pain. Symptoms were quantified using numeric rating scales (NRSs) of pain severity, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Intermittent and Constant Osteoarthritis Pain (ICOAP) and painDETECT questionnaires. Symptoms at other joints were categorised as present/absent. The medial joint line (n = 123, 75%), patellar tendon (n = 62, 38%) and posterior knee (n = 61, 37%) were the most frequently reported pain zones. The most frequent patterns were diffuse (41%), isolated medial (16%), anterior-medial (12%) and medial-posterior (11%) pain. WOMAC and ICOAP scores were higher in the diffuse compared to anterior-medial patterns. Mean PainDETECT scores were higher with both diffuse and medial-posterior pain relative to anterior-medial pain. Only 16% of the cohort indicated isolated medial knee pain, whilst a diffuse pain pattern was most common. People with diffuse knee pain reported more severe pain and physical dysfunction than those with anterior-medial pain. Prevalence of possible/likely neuropathic-like symptoms tended to be more frequent in diffuse and posterior-medial patterns compared to anterior-medial pain.


In-service light path PMD (polarization mode dispersion) monitoring by PMD compensation.
We describe a novel method of measuring PMD (polarization mode dispersion) of an in-service DWDM system by PMD compensation. We successfully demonstrate the method in a 1500-km ultra-long haul DWDM test bed. We further verify the feasibility of the method for in-service light path PMD monitoring in a field trial in a revenue-generating route in Verizon Network, and obtain an accurate PMD value without impacting live network traffic. The discrepancy between the measured and expected PMD values is less than 6% for all cases tested.


Transient floral change and rapid global warming at the Paleocene-Eocene boundary.
Rapid global warming of 5 degrees to 10 degrees C during the Paleocene-Eocene Thermal Maximum (PETM) coincided with major turnover in vertebrate faunas, but previous studies have found little floral change. Plant fossils discovered in Wyoming, United States, show that PETM floras were a mixture of native and migrant lineages and that plant range shifts were large and rapid (occurring within 10,000 years). Floral composition and leaf shape and size suggest that climate warmed by approximately 5 degrees C during the PETM and that precipitation was low early in the event and increased later. Floral response to warming and/or increased atmospheric CO2 during the PETM was comparable in rate and magnitude to that seen in postglacial floras and to the predicted effects of anthropogenic carbon release and climate change on future vegetation.


MOPSA: A microfluidics-optimized particle simulation algorithm.
Computer simulation plays a growing role in the design of microfluidic chips. However, the particle tracers in some existing commercial computational fluid dynamics software are not well suited for accurately simulating the trajectories of particles such as cells, microbeads, and droplets in microfluidic systems. To address this issue, we present a microfluidics-optimized particle simulation algorithm (MOPSA) that simulates the trajectories of cells, droplets, and other particles in microfluidic chips with more lifelike results than particle tracers in existing commercial software. When calculating the velocity of a particle, MOPSA treats the particle as a two-dimensional rigid circular object instead of a single point. MOPSA also checks for unrealistic interactions between particles and channel walls and applies an empirical correcting function to eliminate these errors. To validate the performance of MOPSA, we used it to simulate a variety of important features of microfluidic devices like channel intersections and deterministic lateral displacement (DLD) particle sorter chips. MOPSA successfully predicted that different particle sizes will have different trajectories in six published DLD experiments from three research groups; these DLD chips were used to sort a variety of different cells, particles, and droplets. While some of these particles are not actually rigid or spherical, MOPSA's approximation of these particles as rigid spheres nonetheless resulted in lifelike simulations of the behaviors of these particles (at least for the particle sizes and types shown here). In contrast, existing commercial software failed to replicate these experiments. Finally, to demonstrate that MOPSA can be extended to simulate other properties of particles, we added support for simulating particle density to MOPSA and then used MOPSA to simulate the operation of a microfluidic chip capable of sorting cells by their density. By enabling researchers to accurately simulate the behavior of some types of particles in microfluidic chips before fabricating the chips, MOPSA should accelerate the development of new microfluidic devices for important applications.