Molecular phylogeny and phylogeography of the Australian freshwater fish genus Galaxiella, with an emphasis on dwarf galaxias (G. pusilla).
The freshwater fauna of Southern Australia is primarily restricted to the southwestern and southeastern corners of the continent, and is separated by a large, arid region that is inhospitable to this biota. This geographic phenomenon has attracted considerable interest from biogeographers looking to explain evolutionary diversification in this region. Here, we employed phylogenetic and phylogeographic approaches to evaluate the effect of this barrier on a group of four galaxiid fish species (Galaxiella) endemic to temperate Southern Australia. We also tested if continental shelf width has influenced connectivity among populations during low sea levels when rivers, now isolated, could have been connected. We addressed these questions by sampling each species across its range using multiple molecular markers (mitochondrial cytochrome b sequences, nuclear S7 intron sequences, and 49 allozyme loci). These data also allowed us to assess species boundaries, to refine phylogenetic affinities, and to estimate species ages. Interestingly, we found compelling evidence for cryptic species in G. pusilla, manifesting as allopatric eastern and western taxa. Our combined phylogeny and dating analysis point to an origin for the genus dating to the early Cenozoic, with three of the four species originating during the Oligocene-Miocene. Each Galaxiella species showed high levels of genetic divergences between all but the most proximate populations. Despite extensive drainage connections during recent low sea levels in southeastern Australia, populations of both species within G. pusilla maintained high levels of genetic structure. All populations experienced Late Pleistocene-Holocene population growth, possibly in response to the relaxation of arid conditions after the last glacial maximum. High levels of genetic divergence and the discovery of new cryptic species have important implications for the conservation of this already threatened group of freshwater species.


Decaffeinated coffee and glucose metabolism in young men.
The epidemiological association between coffee drinking and decreased risk of type 2 diabetes is strong. However, caffeinated coffee acutely impairs glucose metabolism. We assessed acute effects of decaffeinated coffee on glucose and insulin levels. This was a randomized, cross-over, placebo-controlled trial of the effects of decaffeinated coffee, caffeinated coffee, and caffeine on glucose, insulin, and glucose-dependent insulinotropic polypeptide (GIP) levels during a 2-h oral glucose tolerance test (OGTT) in 11 young men. Within the first hour of the OGTT, glucose and insulin were higher for decaffeinated coffee than for placebo (P < 0.05). During the whole OGTT, decaffeinated coffee yielded higher insulin than placebo and lower glucose and a higher insulin sensitivity index than caffeine. Changes in GIP could not explain any beverage effects on glucose and insulin. Some types of decaffeinated coffee may acutely impair glucose metabolism but less than caffeine.


Acute Effects of Battle Rope Exercise on Performance, Blood Lactate Levels, Perceived Exertion, and Muscle Soreness in Collegiate Basketball Players.
Chen, WH, Yang, WW, Lee, YH, Wu, HJ, Huang, CF, and Liu, C. Acute effects of battle rope exercise on performance, blood lactate levels, perceived exertion, and muscle soreness in collegiate basketball players. J Strength Cond Res 34(10): 2857-2866, 2020-This study investigated the acute effects of battle rope (BR) exercise on basketball players' performance, blood lactate levels, rating of perceived exertion (RPE), and perceived muscle soreness. Fifteen well-trained Division-I male basketball players underwent the same test procedure at baseline, before BR exercise (30 minutes of rest after the baseline test), and after BR exercise. The 30-minute experimental protocol comprised 6 BR exercises at a work-to-rest ratio of 1:2 (20-second exercise and 40-second rest). Shooting accuracy, basketball chest pass speed, countermovement jump (CMJ) height, blood lactate levels, RPE (Borg Category-Ratio-10 scale), and perceived muscle soreness (visual analog scale, 0-100 mm) were measured in each test. The results indicated no change for any variables between baseline and before BR exercise. After BR exercise, performance decrements (p < 0.05) were recorded in shooting accuracy (16.9%) and basketball chest pass speed (9.1%), but no significant changes were observed for CMJ height. Battle rope exercise caused increases in blood lactate levels (13.6 mmol·L), RPE (9.9), and perceived muscle soreness (upper-limb: 63-67 mm; trunk: 43-68 mm; and lower-limb: 45-52 mm). In conclusion, BR exercise is physically demanding on the upper body, resulting in decreased performance in shooting accuracy and basketball chest pass speed. Battle rope exercise may not be beneficial before a practice or game because it triggers acute exercise-induced performance decrements and fatigue. However, BR exercise may be suitable for basketball training sessions in which the objective is to strengthen technical skills under fatiguing conditions.


Somatic gain-of-function HIF2A mutations in sporadic central nervous system hemangioblastomas.
Central nervous system hemangioblastomas (CNS-HBs) occur sporadically or as a component of von Hippel-Lindau-VHL syndrome. CNS-HBs share some molecular similarities with pheochromocytomas/paragangliomas (PPGLs) and renal cell carcinomas (RCCs). Recently, hypoxia-inducible factors, particularly somatic HIF2A mutations, have been found to play an important role in the pathogenesis of PPGLs. Somatic mutations in HIF2A have been reported in PPGLs associated with polycythemia, which have been reported to also be present in patients with RCCs and HBs. However, whether CNS-HBs is associated with the presence of a HIF2A mutation is currently uknown. We analyzed somatic HIF2A and VHL mutations in a series of 28 sporadic CNS-HBs. We also investigated the expression of HIF target proteins and hypoxia-associated factor (HAF). Two sporadic CNS-HBs were found to have somatic HIF2A mutations. One tumor had 2 HIF2A missense mutations, one of which was previously described in a PPGL (c.1121 T>A, F374Y). The second patient had coexistence of somatic truncated mutations (c.1669 C>T, Q557*) in HIF2A together with a VHL mutation. Neither of the two patients had polycythemia at the time of diagnosis. We demonstrate that the novel truncated mutation in HIF2A (Q557*) affects HIF-2α prolyl hydroxylation with its reduced ubiquitination but intact transcriptional activity, resulting in an activating effect. Both CNS-HB samples showed positive expression of VEGFR2/CA9/Glut1 and HAF. Our data support the unique central role of the VHL/HIF-2α signaling pathway in the molecular pathogenesis of CNS-HBs and show for the first time the presence of HIF2A mutations in sporadic HB.


Molecular-functional studies of adaptive genetic variation in prokaryotes and eukaryotes.
Knowledge of both prokaryotic and eukaryotic organisms is essential to the study of molecular evolution. Their common ancestry mandates that their molecular functions share many aspects of adaptation and constraint, yet their differences in size, ploidy, and structural complexity also give rise to divergent evolutionary options. We explore the interplay of adaptation, constraint, and neutrality in their evolution by the use of genetic variants to probe molecular function in context of molecular structure, metabolic organization, and phenotype-environment interactions. Case studies ranging from bacteria to butterflies, flies, and vertebrates emphasize, among other points: the importance of moving from initial recording of evolutionary pattern variation to studying the processes underlying the patterns, by experiment, reconstructive inference, or both; the complementarity, not conflict, of finding different performance and fitness impacts of natural variants in prokaryotes or eukaryotes, depending on the nature and magnitude of the variants, their locations and roles in pathways, the nature of molecular function affected, and the resulting organismal phenotype-environment interactions leading to selection or its absence; the importance of adaptive functional interaction of different kinds of variants, as in gene expression variants versus variants altering polypeptide properties, or interaction of changes in enzymes' active sites with complementary changes elsewhere that adjust catalytic function in different ways, or coadaptation of different steps' properties in pathways; the power afforded by combining structural and functional analyses of variants with study of the variants' phenotype-environment interactions to understand how molecular changes affect (or fail to affect) adaptive mechanisms "in the wild." Comparative study of prokaryotes and eukaryotes in this multifaceted way promises to deliver both new insights into evolution and a host of new and productive questions about it.


Thermal management and non-reciprocal control of phonon flow via optomechanics.
Engineering phonon transport in physical systems is a subject of interest in the study of materials, and has a crucial role in controlling energy and heat transfer. Of particular interest are non-reciprocal phononic systems, which in direct analogy to electric diodes, provide a directional flow of energy. Here, we propose an engineered nanostructured material, in which tunable non-reciprocal phonon transport is achieved through optomechanical coupling. Our scheme relies on breaking time-reversal symmetry by a spatially varying laser drive, which manipulates low-energy acoustic phonons. Furthermore, we take advantage of developments in the manipulation of high-energy phonons through controlled scattering mechanisms, such as using alloys and introducing disorder. These combined approaches allow us to design an acoustic isolator and a thermal diode. Our proposed device will have potential impact in phonon-based information processing, and heat management in low temperatures.


Occurrence, genotoxicity, and carcinogenicity of regulated and emerging disinfection by-products in drinking water: a review and roadmap for research.
Disinfection by-products (DBPs) are formed when disinfectants (chlorine, ozone, chlorine dioxide, or chloramines) react with naturally occurring organic matter, anthropogenic contaminants, bromide, and iodide during the production of drinking water. Here we review 30 years of research on the occurrence, genotoxicity, and carcinogenicity of 85 DBPs, 11 of which are currently regulated by the U.S., and 74 of which are considered emerging DBPs due to their moderate occurrence levels and/or toxicological properties. These 74 include halonitromethanes, iodo-acids and other unregulated halo-acids, iodo-trihalomethanes (THMs), and other unregulated halomethanes, halofuranones (MX [3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone] and brominated MX DBPs), haloamides, haloacetonitriles, tribromopyrrole, aldehydes, and N-nitrosodimethylamine (NDMA) and other nitrosamines. Alternative disinfection practices result in drinking water from which extracted organic material is less mutagenic than extracts of chlorinated water. However, the levels of many emerging DBPs are increased by alternative disinfectants (primarily ozone or chloramines) compared to chlorination, and many emerging DBPs are more genotoxic than some of the regulated DBPs. Our analysis identified three categories of DBPs of particular interest. Category 1 contains eight DBPs with some or all of the toxicologic characteristics of human carcinogens: four regulated (bromodichloromethane, dichloroacetic acid, dibromoacetic acid, and bromate) and four unregulated DBPs (formaldehyde, acetaldehyde, MX, and NDMA). Categories 2 and 3 contain 43 emerging DBPs that are present at moderate levels (sub- to low-mug/L): category 2 contains 29 of these that are genotoxic (including chloral hydrate and chloroacetaldehyde, which are also a rodent carcinogens); category 3 contains the remaining 14 for which little or no toxicological data are available. In general, the brominated DBPs are both more genotoxic and carcinogenic than are chlorinated compounds, and iodinated DBPs were the most genotoxic of all but have not been tested for carcinogenicity. There were toxicological data gaps for even some of the 11 regulated DBPs, as well as for most of the 74 emerging DBPs. A systematic assessment of DBPs for genotoxicity has been performed for approximately 60 DBPs for DNA damage in mammalian cells and 16 for mutagenicity in Salmonella. A recent epidemiologic study found that much of the risk for bladder cancer associated with drinking water was associated with three factors: THM levels, showering/bathing/swimming (i.e., dermal/inhalation exposure), and genotype (having the GSTT1-1 gene). This finding, along with mechanistic studies, highlights the emerging importance of dermal/inhalation exposure to the THMs, or possibly other DBPs, and the role of genotype for risk for drinking-water-associated bladder cancer. More than 50% of the total organic halogen (TOX) formed by chlorination and more than 50% of the assimilable organic carbon (AOC) formed by ozonation has not been identified chemically. The potential interactions among the 600 identified DBPs in the complex mixture of drinking water to which we are exposed by various routes is not reflected in any of the toxicology studies of individual DBPs. The categories of DBPs described here, the identified data gaps, and the emerging role of dermal/inhalation exposure provide guidance for drinking water and public health research.


Hypothermic aortic arch flush for preservation during exsanguination cardiac arrest of 15 minutes in dogs.
Trauma victims rarely survive cardiac arrest from exsanguination. Survivors may suffer neurologic damage. Our hypothesis was that a hypothermic aortic arch flush of 500 mL of isotonic saline solution at 4 degrees C, compared with 24 degrees C (room temperature), administered at the start of prolonged exsanguination cardiac arrest (CA) would improve functional neurologic outcome in dogs. Seventeen male hunting dogs were prepared under light N2O-halothane anesthesia. The animals were randomized into two groups: group I (n = 9) received 4 degrees C isotonic saline flush and group II (n = 6) received 24 degrees C flush. Two additional dogs received no flush. While spontaneously breathing, the dogs underwent normothermic (tympanic membrane temperature [Ttm] = 37.5 degrees C) exsanguination over 5 minutes to cardiac arrest, assured by electric induction of ventricular fibrillation. After 2 minutes of arrest, the flush was administered over 1 minute into the aortic arch by means of a 13 French balloon-tipped catheter inserted by means of the femoral artery. After 15 minutes of CA, resuscitation was with closed-chest cardiopulmonary bypass, return of shed blood, and defibrillation. For the first 12 hours after CA, core temperature was maintained at 34 degrees C. Mechanical ventilation was continued to 20 hours and intensive care to 72 hours, when final evaluation and perfusion-fixation killing for brain histologic damage scoring were performed. Three dogs in group I were excluded because of extracerebral complications. All 14 dogs that followed protocol survived. During CA, the Ttm decreased to 33.6 +/- 1.2 degrees C in group I and 35.9 +/- 0.4 degrees C in group II (p = 0.002). At 72 hours, in group I, all dogs achieved an overall performance category (OPC) of 1 (normal). In group II, 1 dog was OPC 2 (moderate disability), 3 dogs were OPC 3 (severe disability), and 2 dogs were OPC 4 (coma). Both dogs without flush were OPC 4. Neurologic deficit scores (NDS 0% = normal, 100% = brain death) were 1 +/- 1% in group I and 41 +/- 12% in group II (p < 0.05). The two dogs without flush achieved an NDS of 47% and 59%. Total brain histologic damage scores were 35 +/- 28 in group I and 82 +/- 17 in group II (p < 0.01); and 124 and 200 in the nonflushed dogs. At the start of 15 minutes of exsanguination cardiac arrest in dogs, hypothermic aortic arch flush allows resuscitation to survival with normal neurologic function and histologically almost clean brains.


Mapping the Multiple Graded Contributions of the Anterior Temporal Lobe Representational Hub to Abstract and Social Concepts: Evidence from Distortion-corrected fMRI.
A growing body of recent convergent evidence indicates that the anterior temporal lobe (ATL) has connectivity-derived graded differences in semantic function: the ventrolateral region appears to be the transmodal, omni-category center-point of the hub whilst secondary contributions come from the peripheries of the hub in a manner that reflects their differential connectivity to different input/output modalities. One of the key challenges for this neurocognitive theory is how different types of concept, especially those with less reliance upon external sensory experience (such as abstract and social concepts), are coded across the graded ATL hub. We were able to answer this key question by using distortion-corrected fMRI to detect functional activations across the entire ATL region and thus to map the neural basis of social and psycholinguistically-matched abstract concepts. Both types of concept engaged a core left-hemisphere semantic network, including the ventrolateral ATL, prefrontal regions and posterior MTG. Additionally, we replicated previous findings of weaker differential activation of the superior and polar ATL for the processing of social stimuli, in addition to the stronger, omni-category activation observed in the vATL. These results are compatible with the view of the ATL as a graded transmodal substrate for the representation of coherent concepts.


Outcome of antenatally diagnosed abdominal wall defects.
To examine the natural history and detailed outcome of antenatally diagnosed abdominal wall defects. This was a retrospective review of the antenatal reports, pediatric surgery records and subsequent follow-up information of all cases of omphalocele and gastroschisis diagnosed in a 10-year period in our tertiary referral center. There were 109 cases of abdominal wall defects, including omphalocele in 67 cases and gastroschisis in 42 cases. Of the 67 cases of omphalocele there were 26 (39%) with chromosomal abnormalities and 22 (33%) underwent termination of pregnancy, mainly for associated structural abnormalities. Of the ongoing 19 cases there were five (26%) in-utero deaths, 12 (63%) survivors and two (11%) neonatal deaths, both associated with prematurity. Excluding chromosomal abnormalities, the survival rate in isolated omphalocele was 7/16 (44%) whilst it was 5/25 (20%) in those with associated abnormalities. Gastroschisis was isolated in 40 (95%) cases. Among these 40 isolated cases there were two (5%) terminations. Of the 38 ongoing cases, there were two (5%) in-utero deaths, and 36 (95%) live births. Four of the 36 liveborn infants (11%) died in the postoperative period owing to complications of small bowel atresia. Although only 18% of infants with antenatally diagnosed omphalocele were alive in the neonatal period, postoperative morbidity was low. The majority (90%) of fetuses with antenatally diagnosed gastroschisis survived to delivery, but the mortality in affected newborns was 11%.


Knee arthroplasties have similar results in high- and low-activity patients.
With increased patient demand to return to active lifestyles, total knee arthroplasties have evolved from primarily providing pain relief to allowing for increased function and mobility. We evaluated the influence of activity on the outcome of total knee arthroplasties. The overall satisfaction, rate of revision, and clinical and radiographic results for high-activity patients were compared with a matched group of low-activity patients at a minimum followup of 4 years (mean, 7 years; range, 4-14 years). There were 22 men and 35 women (72 knees) in each group. High-activity patients fared as well as their low-activity counterparts with no differences in clinical outcomes. At the time of last followup, mean Knee Society objective scores were 95 points (range, 70-100 points) and 96 points (range, 80-100 points) for the high-activity and low-activity groups, respectively. The high-activity group had one clinical failure, and neither group had any revisions. The groups had similar radiographic outcomes with no progressive radiolucencies and no evidence of osteolysis. While we await long-term results, these results suggest that low- to moderate-impact sports activities had no effect on the clinical and/or radiographic outcomes of total knee arthroplasties at mid-term followup.


Impact of influenza exposure on rates of hospital admissions and physician visits because of respiratory illness among pregnant women.
Excess deaths have occurred among pregnant women during influenza pandemics, but the impact of influenza during nonpandemic years is unclear. We evaluated the impact of exposure during nonpandemic influenza seasons on the rates of hospital admissions and physician visits because of respiratory illness among pregnant women. We conducted a 13-year (1990-2002) population-based cohort study involving pregnant women in Nova Scotia. We compared rates of hospital admissions and physician office visits because of respiratory illness during the influenza season in each trimester of pregnancy with rates during the influenza season in the year before pregnancy and with rates in non-influenza seasons. Poisson regression analyses were performed to estimate rate ratios and 95% confidence intervals (CIs). Of 134,188 pregnant women in the study cohort, 510 (0.4%) were admitted to hospital because of a respiratory illness during pregnancy and 33,775 (25.2%) visited their physician for the same reason during pregnancy. During the influenza seasons, the rate ratio of hospital admissions in the third trimester compared with admissions in the year before pregnancy was 7.9 (95% CI 5.0-12.5) among women with comorbidities and 5.1 (95% CI 3.6-7.3) among those without comorbidities. The rate of hospital admissions in the third trimester among women without comorbidities was 7.4 per 10,000 woman-months during the influenza season, compared with 5.4 and 3.1 per 10,000 woman-months during the peri-and non-influenza seasons respectively. Corresponding rates among women with comorbidities were 44.9, 9.3 and 18.9 per 10,000 woman-months. Only 6.7% of women with comorbidities had received influenza immunization. Our data support the recommendation that pregnant women with comorbidities should receive influenza vaccination regardless of their stage of pregnancy during the influenza season. Since hospital admissions because of respiratory illness during the influenza season were also increased among pregnant women without comorbidities, all pregnant women are likely to benefit from influenza vaccination.


Arteriovenous malformation (AVM) of the maxilla complicated by a local pathogen infection.
Arteriovenous malformations (AVM) are usually observed on the skin, but they can also rarely affect the visceral organs and the maxillofacial area. In the maxillofacial area, these lesions can spread and they are potentially dangerous, as a biopsy or even a simple tooth extraction can lead to a catastrophic haemorrhage that may eventually cause death. There are several indications for the treatment, including age, size and type of vascular malformation. The purpose of this report is to describe an AVM on the maxilla, first diagnosed as an epulis, which has been treated for several years as a chronic local periodontitis. The lesion was finally stabilised after an antibiotic therapy. Improved knowledge of the development of these lesions and collaboration between dentists, radiologists and surgeons are necessary to improve the diagnosis and treatment of vascular malformations.


Schizophrenia disorders, substance abuse and prior offending in a sequential series of 435 homicides.
To examine the relationship between committing homicide, the presence of schizophrenia, substance misuse and past criminality. The study employed a data linkage design, using contacts recorded on two statewide databases, one of which recorded public mental health services contacts and the second of which recorded contacts with the police. The estimated rates of schizophrenia disorders, substance abuse and criminal convictions found among a population of 435 homicide offenders were contrasted with estimated rates in two composite comparison samples. Of the 435 offenders, 38 (8.7%) had been diagnosed with a schizophrenia disorder, which was RR 13.11 (95% CI 9.14-18.80) times more likely than a comparison sample. Rates of known substance abuse between homicide offenders with and without schizophrenia and community-dwelling residents with schizophrenia did not differ significantly. However, these rates were higher than those found in the general community. A similar pattern emerged for comparisons regarding offending histories between these same groups. The association between homicidal violence and having a schizophrenia disorder cannot be explained away simply on the basis of either comorbid substance abuse or prior criminal offending.


A comparison of acute and long-term management of stroke patients in Barbados and South London.
To compare health care utilisation between stroke patients living in a middle-income country with similar patients in a high-income country in terms of the type and amount of health care received following a stroke. Data were collected from the population-based South London Stroke Register (SLSR) and the Barbados Register of Strokes (BROS) from January 2001 to December 2004. Differences in management and diagnostic procedures used in the acute phase were adjusted for age, sex, ethnic group, living conditions pre-stroke and socio-economic status by multivariable logistic regression. Comparison of subsequent management was made for 3 months and 1 year post-stroke. Patients in BROS were less likely to be admitted to a hospital ward (OR 0.22; 95% CI 0.13-0.37), but the difference for the lower use of brain scans in BROS was smaller (OR 0.62; 95% CI 0.25-1.52). Additional adjustment for stroke severity (Glasgow Coma Score) showed that BROS patients were more likely to have a swallow test on admission (OR 2.95; 95% CI 1.17-7.45). BROS patients were less likely to be in nursing care at 3 months (OR 0.37; 95% CI 0.17-0.81), and less likely to be receiving speech and language therapy at 3 months (OR 0.10; 95% CI 0.03-0.33) and 1 year (OR 0.05; 95% CI 0.00-0.55). The lower use of hospital admission and nursing care at 3 months suggests that in Barbados, family and friends take greater responsibility for patient care around the time of the stroke and in the medium term thereafter.


Simultaneous measurement of refractive index and wedge angle of optical windows using Fizeau interferometry and a cyclic path optical configuration.
We present a new technique for the simultaneous measurement of refractive index and wedge angle of optical windows using Fizeau interferometry and a cyclic path optical configuration (CPOC). Two laterally separated beams are obtained from an expanded collimated beam using an aperture containing two rectangular openings. The test wedge plate is placed in one of the two separated beams. Using CPOC, these two beams are made to overlap and interfere, producing interference fringes in the overlapping region. The beams reflected from the front and back surfaces of the test wedge plate interfere and produce Fizeau fringes. The refractive index is related to the spacing of the above two beam fringes. The wedge angle is determined from the evaluated values of the refractive index and Fizeau fringe spacing. The results obtained for a BK-7 optical window are presented.


Protocol for the OUTREACH trial: a randomised trial comparing delivery of cancer systemic therapy in three different settings: patient's home, GP surgery and hospital day unit.
The national Cancer Reform Strategy recommends delivering care closer to home whenever possible. Cancer drug treatment has traditionally been administered to patients in specialist hospital-based facilities. Technological developments mean that nowadays, most treatment can be delivered in the out-patient setting. Increasing demand, care quality improvements and patient choice have stimulated interest in delivering some treatment to patients in the community, however, formal evaluation of delivering cancer treatment in different community settings is lacking. This randomised trial compares delivery of cancer treatment in the hospital with delivery in two different community settings: the patient's home and general practice (GP) surgeries. Patients due to receive a minimum 12 week course of standard intravenous cancer treatment at two hospitals in the Anglia Cancer Network are randomised on a 1:1:1 basis to receive treatment in the hospital day unit (control arm), or their own home, or their choice of one of three neighbouring GP surgeries. Overall patient care, treatment prescribing and clinical review is undertaken according to standard local practice. All treatment is dispensed by the local hospital pharmacy and treatment is delivered by the hospital chemotherapy nurses. At four time points during the 12 week study period, information is collected from patients, nursing staff, primary and secondary care teams to address the primary end point, patient-perceived benefits (using the emotional function domain of the EORTC QLQC30 patient questionnaire), as well as secondary end points: patient satisfaction, safety and health economics. The Outreach trial is the first randomised controlled trial conducted which compares delivery of out-patient based intravenous cancer treatment in two different community settings with standard hospital based treatment. Results of this study may better inform all key stakeholders regarding potential costs and benefits of transferring clinical services from hospital to the community. ISRCTN: ISRCTN66219681.


FKBP12-Dependent Inhibition of Calcineurin Mediates Immunosuppressive Antifungal Drug Action in Malassezia.
The genus Malassezia includes yeasts that are commonly found on the skin or hair of animals and humans as commensals and are associated with a number of skin disorders. We have previously developed an Agrobacterium tumefaciens transformation system effective for both targeted gene deletion and insertional mutagenesis in Malassezia furfur and M. sympodialis In the present study, these molecular resources were applied to characterize the immunophilin FKBP12 as the target of tacrolimus (FK506), ascomycin, and pimecrolimus, which are calcineurin inhibitors that are used as alternatives to corticosteroids in the treatment of inflammatory skin disorders such as those associated with Malassezia species. While M. furfur and M. sympodialis showed in vitro sensitivity to these agents, fkb1Δ mutants displayed full resistance to all three of them, confirming that FKBP12 is the target of these calcineurin inhibitors and is essential for their activity. We found that calcineurin inhibitors act additively with fluconazole through an FKBP12-dependent mechanism. Spontaneous M. sympodialis isolates resistant to calcineurin inhibitors had mutations in the gene encoding FKBP12 in regions predicted to affect the interactions between FKBP12 and FK506 based on structural modeling. Due to the presence of homopolymer nucleotide repeats in the gene encoding FKBP12, an msh2Δ hypermutator of M. sympodialis was engineered and exhibited an increase of more than 20-fold in the rate of emergence of resistance to FK506 compared to that of the wild-type strain, with the majority of the mutations found in these repeats.IMPORTANCEMalassezia species are the most abundant fungal components of the mammalian and human skin microbiome. Although they belong to the natural skin commensal flora of humans, they are also associated with a variety of clinical skin disorders. The standard treatment for Malassezia-associated inflammatory skin infections is topical corticosteroids, although their use has adverse side effects and is not recommended for long treatment periods. Calcineurin inhibitors have been proposed as a suitable alternative to treat patients affected by skin lesions caused by Malassezia Although calcineurin inhibitors are well-known as immunosuppressive drugs, they are also characterized by potent antimicrobial activity. In the present study, we investigated the mechanism of action of FK506 (tacrolimus), ascomycin (FK520), and pimecrolimus in M. furfur and M. sympodialis and found that the conserved immunophilin FKBP12 is the target of these drugs with which it forms a complex that directly binds calcineurin and inhibits its signaling activity. We found that FKBP12 is also required for the additive activity of calcineurin inhibitors with fluconazole. Furthermore, the increasing natural occurrence in fungal pathogen populations of mutator strains poses a high risk for the rapid emergence of drug resistance and adaptation to host defense. This led us to generate an engineered hypermutator msh2Δ mutant strain of M. sympodialis and genetically evaluate mutational events resulting in a substantially increased rate of resistance to FK506 compared to that of the wild type. Our study paves the way for the novel clinical use of calcineurin inhibitors with lower immunosuppressive activity that could be used clinically to treat a broad range of fungal infections, including skin disorders caused by Malassezia.


Multiple mechanisms participate in the generation of diversity of human H chain CDR3 regions.
The area of highest variability in the antibody-combining site is the third complementarity determining region or CDR3. Based on our preliminary observation of the tremendous variability of this region in the human system we have studied the potential of CDR3 regions for generating diversity in the human B cell repertoire. To this end we generated CDR3-specific cDNA libraries from tissues collected at several stages of human development. Detailed computer analysis of more than 500 sequences reveals that human CDR3 region have the potential to generate more than 10(14) different peptides. The mechanisms responsible for this diversity include rearrangement by inversion, D-D fusion, gene conversion and the frequent utilization of the recently described DIR genes. The specific recombination mechanisms which may explain aberrant rearrangements as well as differences between fetal and adult repertoires are discussed in detail.


Sleep disturbances in patients with chronic heart failure and their holistic consequences-what different care actions can be implemented?
Sleep disturbances are prevalent among elderly, especially among those with chronic heart failure (CHF) and can affect all dimensions of quality of life (QOL) negatively. To describe the most common causes leading to sleep disturbances in patients with CHF, their consequences from a holistic perspective and different care actions that can be implemented. MEDLINE and CINAHL databases were searched from 1989 to July 2004. Sleep disordered breathing (SDB), and insomnia were the most common causes for sleep disturbances and occurs in 45--82% (SDB) and one-third (insomnia) of all patients with CHF. SDB cause a disturbed sleep structure with frequent awakenings, as well as several adverse effects on the cardiovascular system causing increased morbidity and mortality. Insomnia, caused by anxiety, an unknown life situation in relation to the debut of CHF, or symptoms/deteriorations of CHF can lead to negative effects on all aspects of QOL, as well as daytime sleepiness. The high prevalence of sleep disturbances and their holistic consequences should be taken into account when nurses asses and plan the care for patients with CHF. Randomized studies with large sample sizes evaluating non-pharmacological nursing interventions that improve sleep are needed.


Comparative phylogenetic histories of two louse genera found on Catharus thrushes and other birds.
The louse genera Brueelia (Ischnocera) and Myrsidea (Amblycera) are broadly codistributed on songbirds (Passeriformes), but differ in a variety of life history characteristics. We used mitochondrial and nuclear DNA sequences to assess levels of genetic divergence and reconstruct phylogenies of these 2 genera, focusing especially on Catharus thrushes in North America. We then qualitatively compared the phylogenies and levels of divergence within these 2 genera of codistributed parasites. Neither Brueelia nor Myrsidea appears to cospeciate with Catharus thrushes or passerine birds in general. The Myrsidea phylogeny exhibits significant levels of biogeographic structure, whereas the Brueelia phylogeny does not. Myrsidea and Brueelia also differ in their levels of intra-generic genetic divergence, with Myrsidea showing higher levels of genetic divergence and host specificity than Brueelia. Our genetic data support traditional morphology-based taxonomy in several instances in which the same species of Brueelia has been reported on multiple host taxa, e.g., all migrant Catharus spp. carry B. antiqua, with little haplotype divergence. Myrsidea found on each Catharus sp. are in general genetically distinct, except for M. incerta, which parasitizes both Catharus ustulatus and Catharus minimus. The strong biogeographic signal in the Myrsidea phylogeny and higher relative levels of host specificity of Myrsidea spp. suggest that infrequent host-switching, followed by speciation, is shaping the evolutionary history of this group. In contrast, the relatively lower host specificity of Brueelia spp. suggests that host-switching, combined with more frequent ongoing dispersal, has been more important in the evolutionary history of Brueelia.


Matrix metalloproteinase expression in human retinal microvascular cells.
The degree of hyperglycemia correlates with the development of diabetic retinopathy. We investigated the effect of glucose on the expression of matrix metalloproteinase (MMP)-2 and MMP-9 (72-kDa and 92-kDa type IV collagenases, respectively) by human retinal microvascular endothelial cells (HRECs). Cultured HRECs from nondiabetic and diabetic donors were exposed to 5 or 30 mmol/l glucose. Using gelatin zymography, conditioned medium (CM) from all cultures revealed a gelatinolytic band migrating at 65 kDa (representing the proform of MMP-2 that runs at 72 kDa under reducing conditions). This band was unchanged by glucose exposure or the disease state of the donors. CM from nondiabetic HREC cultures demonstrated an additional proteolytic activity migrating at 90 kDa when cells were exposed to 30 mmol/l glucose, but not when they were exposed to 5 mmol/l glucose. This same activity was seen in CM from HREC cultures of diabetic origin in the presence of both 5 and 30 mmol/l glucose. Western analysis confirmed the identity of the 65-kDa band as MMP-2. The anomalous activity at 90 kDa was identified as MMP-2 associated and co-migrating with a fibronectin fragment. Competition-based reverse transcription-polymerase chain reaction revealed that nondiabetic and diabetic HRECs expressed constitutively mRNA for MMP-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, and fibronectin. After exposure to 5 or 30 mmol/l glucose, no changes were detected in mRNA levels in MMP-2 or MMP-9, their inhibitors TIMP-1 and TIMP-2, or fibronectin in either nondiabetic or diabetic HREC cultures. These results support the notion that modulation of MMP function by extracellular matrix components occurs in response to glucose and may be relevant to the development of diabetic retinopathy.


Screening of Natural Compounds as P-Glycoprotein Inhibitors against Multidrug Resistance.
Multidrug resistance (MDR) is a common problem when fighting cancer with chemotherapy. P-glycoprotein (P-gp, or MDR1) is an active pump responsible for the efflux of xenobiotics out of the cell, including anti-cancer drugs. It is a validated target against MDR. No crystal structure of the human P-gp is available to date, and only recently several cryo-EM structures have been solved. In this paper, we present a comprehensive computational approach that includes constructing the full-length three-dimensional structure of the human P-gp and its refinement using molecular dynamics. We assessed its flexibility and conformational diversity, compiling a dynamical ensemble that was used to dock a set of lignan compounds, previously reported as active P-gp inhibitors, and disclose their binding modes. Based on the statistical analysis of the docking results, we selected a system for performing the structure-based virtual screening of new potential P-gp inhibitors. We tested the method on a library of 87 natural flavonoids described in the literature, and 10 of those were experimentally assayed. The results reproduced the theoretical predictions only partially due to various possible factors. However, at least two of the predicted natural flavonoids were demonstrated to be effective P-gp inhibitors. They were able to increase the accumulation of doxorubicin inside the human promyelocytic leukemia HL60/MDR cells overexpressing P-gp and potentiate the antiproliferative activity of this anti-cancer drug.


Biomimetic noncationic lipid nanoparticles for mRNA delivery.
Although the tremendous progress has been made for mRNA delivery based on classical cationic carriers, the excess cationic charge density of lipids was necessary to compress mRNA through electrostatic interaction, and with it comes inevitably adverse events including the highly inflammatory and cytotoxic effects. How to develop the disruptive technologies to overcome cationic nature of lipids remains a major challenge for safe and efficient mRNA delivery. Here, we prepared noncationic thiourea lipids nanoparticles (NC-TNP) to compress mRNA by strong hydrogen bonds interaction between thiourea groups of NC-TNP and the phosphate groups of mRNA, abandoning the hidebound and traditional electrostatic force to construct mRNA-cationic lipids formulation. NC-TNP was a delivery system for mRNA with simple, convenient, and repeatable preparation technology and showed negligible inflammatory and cytotoxicity side effects. Furthermore, we found that NC-TNP could escape the recycling pathway to inhibit the egress of internalized nanoparticles from the intracellular compartment to the extracellular milieu which was a common fact in mRNA-LNP (lipid nanoparticles) formulation. Therefore, NC-TNP-encapsulated mRNA showed higher gene transfection efficiency in vitro and in vivo than mRNA-LNP formulation. Unexpectedly, NC-TNP showed spleen targeting delivery ability with higher accumulation ratio (spleen/liver), compared with traditional LNP. Spleen-targeting NC-TNP with mRNA exhibited high mRNA-encoded antigen expression in spleen and elicited robust immune responses. Overall, our work establishes a proof of concept for the construction of a noncationic system for mRNA delivery with good inflammatory safety profiles, high gene transfection efficiency, and spleen-targeting delivery to induce permanent and robust humoral and cell-mediated immunity for disease treatments.


Heritability of behavioural tolerance to high CO2 in a coral reef fish is masked by nonadaptive phenotypic plasticity.
Previous studies have demonstrated limited potential for acclimation of adversely affected olfactory behaviours in reef fishes under elevated CO 2, indicating that genetic adaptation will be required to maintain behavioural performance in the future. Adaptation depends on the presence of heritable phenotypic variation in the trait, which may differ between populations and environments. We used parent-offspring regressions to estimate the heritability (h2) of variation in behavioural tolerance to high CO 2 (754 μatm) in both field-collected and laboratory-reared families of Acanthochromis polyacanthus. Tolerance to elevated CO 2 was measured by determining the behavioural response of individuals to chemical alarm cues. Both populations exhibited high heritability of olfactory behaviour phenotype (father-mid-offspring h2 = 0.56 & 0.65, respectively) when offspring were acutely exposed to high CO 2 for 4 days. However, there was no heritability in the behavioural phenotype when juveniles were chronically exposed to high CO 2 for 6 weeks in the laboratory-reared families. Parental exposure to high CO 2 during the breeding season did not alter this relationship between heritability and length of juvenile exposure to high CO 2. These results demonstrate that variation in behavioural tolerance to high CO 2 is heritable, but adaptive potential may be constrained by a loss of phenotypic variation when juveniles permanently experience a high-CO 2 environment, as will occur with rising CO 2 levels in the ocean.


Incidence of retinopathy of prematurity in infants born before 27 weeks' gestation in Sweden.
To determine the incidence of retinopathy of prematurity (ROP) in extremely preterm infants born before 27 weeks' gestation in Sweden during a 3-year period. A national, prospective, population-based study was performed in Sweden from April 1, 2004, to March 31, 2007. The ophthalmologic part of the study was separately organized, and screening for ROP was performed beginning postnatal week 5. The criteria for the treatment of ROP agreed with the recommendations of the Early Treatment for Retinopathy of Prematurity Cooperative Group. During the study, 506 of 707 live-born infants survived until the first eye examination. Of these, 368 (72.7%) had ROP: 37.9% had mild ROP and 34.8% had severe ROP. Ninety-nine infants (19.6%) were treated. Gestational age at birth was a stronger predictor of ROP than was birth weight. A log-linear relationship between severe ROP and gestational age at birth was found in the present cohort, and the risk of ROP was reduced by 50% for each week of increase in gestational age at birth. Today, extremely preterm infants are surviving, and this population-based study with ROP as a primary outcome shows a higher incidence of this condition than in previously reported national cohorts.


Assessing Variations in Areal Organization for the Intrinsic Brain: From Fingerprints to Reliability.
Resting state fMRI (R-fMRI) is a powerful in-vivo tool for examining the functional architecture of the human brain. Recent studies have demonstrated the ability to characterize transitions between functionally distinct cortical areas through the mapping of gradients in intrinsic functional connectivity (iFC) profiles. To date, this novel approach has primarily been applied to iFC profiles averaged across groups of individuals, or in one case, a single individual scanned multiple times. Here, we used a publically available R-fMRI dataset, in which 30 healthy participants were scanned 10 times (10 min per session), to investigate differences in full-brain transition profiles (i.e., gradient maps, edge maps) across individuals, and their reliability. 10-min R-fMRI scans were sufficient to achieve high accuracies in efforts to "fingerprint" individuals based upon full-brain transition profiles. Regarding test-retest reliability, the image-wise intraclass correlation coefficient (ICC) was moderate, and vertex-level ICC varied depending on region; larger durations of data yielded higher reliability scores universally. Initial application of gradient-based methodologies to a recently published dataset obtained from twins suggested inter-individual variation in areal profiles might have genetic and familial origins. Overall, these results illustrate the utility of gradient-based iFC approaches for studying inter-individual variation in brain function.


The Challenge of Measuring UK Wealth Inequality in the 2000s.
The concentration of personal wealth is now receiving a great deal of attention - after having been neglected for many years. One reason is the growing recognition that, in seeking explanations for rising income inequality, we need to look not only at wages and earned income but also at income from capital, particularly at the top of the distribution. In this paper, we use evidence from existing data sources to attempt to answer three questions: (i) What is the share of total personal wealth that is owned by the top 1 per cent, or the top 0.1 per cent? (ii) Is wealth much more unequally distributed than income? (iii) Is the concentration of wealth at the top increasing over time? The main conclusion of the paper is that the evidence about the UK concentration of wealth post-2000 is seriously incomplete and significant investment in a variety of sources is necessary if we are to provide satisfactory answers to the three questions.


Quality of motivational interviewing matters: the effect on participation in health-promotion activities in a cluster randomized controlled trial.
Objectives This study aims to evaluate the effectiveness of a "blended" workplace health-promotion intervention, alongside identification of key components beneficial for future implementation strategies. Methods Within a cluster randomized controlled trial, 491 employees at increased risk of cardiovascular disease were allocated to the limited (N=217; 9 clusters) or extensive (N=274; 8 clusters) intervention. The extensive intervention consisted of motivational interviewing (MI) within the framework of a web-based health risk assessment (HRA), a blended care approach. The limited intervention received solely the web-based HRA. Occupational health physicians (N=21) within three organizations delivered the intervention. Implementation components investigated included: HRA (reach and participation), newsletters (percentage read), and MI sessions (number and quality). MI quality was determined by scoring audiotaped MI sessions, using the MI treatment integrity code. After 6 and 12 months, effects on participation in health-promotion activities and its associations with components of implementation were determined by mixed-effects models. Results Over 80% of employees participated in health-promotion activities, with an additional 8% in the extensive compared to the limited group. In the extensive intervention, those with more or better quality MI sessions were more likely to participate in health-promotion activities. Increased MI quality was associated with sustained participation. Conclusions This study suggests that participation in health-promotion activities can be increased by adding MI to a web-based approach and improving the quality of the MI delivered. Interventions with MI should include optimized delivery and quality of MI sessions.


Assessing the statistical significance of the achieved classification error of classifiers constructed using serum peptide profiles, and a prescription for random sampling repeated studies for massive high-throughput genomic and proteomic studies.
Peptide profiles generated using SELDI/MALDI time of flight mass spectrometry provide a promising source of patient-specific information with high potential impact on the early detection and classification of cancer and other diseases. The new profiling technology comes, however, with numerous challenges and concerns. Particularly important are concerns of reproducibility of classification results and their significance. In this work we describe a computational validation framework, called PACE (Permutation-Achieved Classification Error), that lets us assess, for a given classification model, the significance of the Achieved Classification Error (ACE) on the profile data. The framework compares the performance statistic of the classifier on true data samples and checks if these are consistent with the behavior of the classifier on the same data with randomly reassigned class labels. A statistically significant ACE increases our belief that a discriminative signal was found in the data. The advantage of PACE analysis is that it can be easily combined with any classification model and is relatively easy to interpret. PACE analysis does not protect researchers against confounding in the experimental design, or other sources of systematic or random error. We use PACE analysis to assess significance of classification results we have achieved on a number of published data sets. The results show that many of these datasets indeed possess a signal that leads to a statistically significant ACE.


Bergenin, a PPARγ agonist, inhibits Th17 differentiation and subsequent neutrophilic asthma by preventing GLS1-dependent glutaminolysis.
Bergenin is a natural PPARγ agonist that can prevent neutrophil aggregation, and often be used in clinics for treating respiratory diseases. Recent data show that Th17 cells are important for neutrophil aggregation and asthma through secreting IL-17A. In this study, we investigated the effects of bergenin on Th17 differentiation in vitro and subsequent neutrophilic asthma in mice. Naïve T cells isolated from mouse mesenteric lymph nodes were treated with IL-23, TGF-β, and IL-6 to induce Th17 differentiation. We showed that in naïve T cells under Th17-polarizing condition, the addition of bergenin (3, 10, 30 μM) concentration-dependently decreased the percentage of CD4+ IL-17A+ T cells and mRNA expression of specific transcription factor RORγt, and function-related factors IL-17A/F, IL-21, and IL-22, but did not affect the cell vitality and apoptosis. Furthermore, bergenin treatment prevented GLS1-dependent glutaminolysis in the progress of Th17 differentiation, slightly affected the levels of SLC1A5, SLC38A1, GLUD1, GOT1, and GPT2. Glutamine deprivation, the addition of glutamate (1 mM), α-ketoglutarate (1 mM), or GLS1 plasmid all significantly attenuated the above-mentioned actions of bergenin. Besides, we demonstrated that bergenin (3, 10, and 30 μM) concentration-dependently activated PPARγ in naïve T cells, whereas PPARγ antagonist GW9662 and siPPARγ abolished bergenin-caused inhibition on glutaminolysis and Th17 differentiation. Furthermore, we revealed that bergenin inhibited glutaminolysis by regulating the level of CDK1, phosphorylation and degradation of Cdh1, and APC/C-Cdh1-mediated ubiquitin-proteasomal degradation of GLS1 after activating PPARγ. We demonstrated a correlation existing among bergenin-affected GLS1-dependent glutaminolysis, PPARγ, "CDK1-APC/C-Cdh1" signaling, and Th17 differentiation. Finally, the therapeutic effect and mechanisms for bergenin-inhibited Th17 responses and neutrophilic asthma were confirmed in a mouse model of neutrophilic asthma by administration of GW9662 or GLS1 overexpression plasmid in vivo. In conclusion, bergenin repressed Th17 differentiation and then alleviated neutrophilic asthma in mice by inhibiting GLS1-dependent glutaminolysis via regulating the "CDK1-APC/C-Cdh1" signaling after activating PPARγ.


Effect of topical beta blockers on corneal epithelial wound healing in the rabbit.
Topical beta adrenergic receptor blocking agents (beta blockers) are among the most frequently prescribed ophthalmic drugs. It has been suggested that some of these agents have a toxic effect on the corneal epithelium. In the present study, four beta blockers in common therapeutic concentrations, as well as their vehicles, were applied to rabbit corneas that had undergone mechanical removal of epithelium from a 6 mm diameter corneal wound. The tested drugs (0.25% timolol, 0.25% levobunolol, 0.25% betaxolol, and 0.3% metipranolol) were found significantly to accelerate wound closure, compared with saline treatment in controls. Eyes treated with two of the vehicles (betaxolol vehicle and metipranolol vehicle) also demonstrated more rapid healing than controls, but the magnitude of the effect was not as great as that seen with the drugs. Only the beta blockers were responsible for wound closure before 60 hours, whereas the saline treated controls and vehicle treated eyes required longer times for wound closure. In this model of wound healing, beta blockers appear to have no deleterious effect on corneal epithelial wound healing.


Perspective: Coarse-grained models for biomolecular systems.
By focusing on essential features, while averaging over less important details, coarse-grained (CG) models provide significant computational and conceptual advantages with respect to more detailed models. Consequently, despite dramatic advances in computational methodologies and resources, CG models enjoy surging popularity and are becoming increasingly equal partners to atomically detailed models. This perspective surveys the rapidly developing landscape of CG models for biomolecular systems. In particular, this review seeks to provide a balanced, coherent, and unified presentation of several distinct approaches for developing CG models, including top-down, network-based, native-centric, knowledge-based, and bottom-up modeling strategies. The review summarizes their basic philosophies, theoretical foundations, typical applications, and recent developments. Additionally, the review identifies fundamental inter-relationships among the diverse approaches and discusses outstanding challenges in the field. When carefully applied and assessed, current CG models provide highly efficient means for investigating the biological consequences of basic physicochemical principles. Moreover, rigorous bottom-up approaches hold great promise for further improving the accuracy and scope of CG models for biomolecular systems.


A Fluorescent Readout for the Oxidation State of Electron Transporting Proteins in Cell Free Settings.
Pathways involving sequential electron transfer between multiple proteins are ubiquitous in nature. Here, we demonstrate a new class of fluorescent protein-based reporters for monitoring electron transport through such multistage cascades, specifically those involving ferredoxin-like electron transporters. We created protein fusions between mammalian Adrenodoxin (Adx) and plant Ferredoxin (Fdx) with fluorescent proteins of different colors and found that the fluorescence of such fusions is highly sensitive to the redox state of the electron transporter. The increase in fluorescence from the oxidized to the reduced state was inversely proportional to the linker length between the fusion partners. We first used our approach to quantitatively characterize electron transfer from NADPH through Adrenodoxin Reductase (AdR) to Adrenodoxin (Adx). Our data allowed us to build a detailed mathematical model of this mitochondrial electron transfer chain and validate previously proposed mechanisms. Then, we showed that an Adx-GFP fusion could serve as a sensor for the activity of bacterial Type I Cytochrome P450s (CYPs), a very large class of enzymes with important roles in biotechnology. We further showed that fluorescence of a direct fusion between CYP and GFP was sensitive to CYP activity, suggesting that our approach is applicable to an even broader class of proteins, which undergo a redox state change during their work cycle.


Subcellular Localization of Survivin Determines Its Function in Cardiomyocytes.
Rationale: Reducing cardiomyocyte death and enhancing their proliferation after myocardial infarction is perhaps the single largest challenge for cardiac tissue regeneration. Survivin (SVV) is the smallest member of the inhibitor of apoptosis (IAP) family but plays two important roles; inhibiting caspase-9 activation in the intrinsic apoptosis pathway, and regulating microtubule dynamics and chromosome segregation during cell division. Genetic depletion of cardiac SVV leads to incomplete cardiomyocyte division and abnormal heart development. However, the function of SVV in adult hearts after myocardial infarction remains unclear. Methods: A homozygous inducible cardiomyocyte-specific SVV knockout transgenic mouse model was established through crossbreeding SVVflox/flox and αMHC-MCM transgenic mice. Adult mice received consecutive intraperitoneal injection of tamoxifen to induce genetic removal of SVV in cardiomyocytes. A SVV overexpressing model was established via local delivery of SVV in wild-type mouse hearts. Results: We found that 30.82% of cardiomyocytes in the peri-infarct region of SVV knockout mice were apoptotic, significantly higher than the 22.18% in control mice. In addition, ejection fraction was 29.00±0.40% in knockout mice compared to 38.04±0.50% in control mice 21 days after myocardial infarction. On the contrary, locally overexpressing SVV in the heart improved cardiac functions. Unexpectedly, we found that altering the subcellular localization of SVV overexpression produced different outcomes. Overexpression of SVV in the cytoplasm decreased cardiomyocyte apoptosis, whereas overexpression of SVV in the nucleus enhanced cardiac regeneration. The ejection fraction of mice overexpressing SVV was 36.58±0.91%, significantly higher than 28.18±1.70% in the GFP control group. Apoptotic cardiomyocytes were only 4.63% in mouse overexpressing cytosolic SVV, compared to 9.31% in the GFP group, and activation of caspase-3 was also reduced. Moreover, mice overexpressing NLS-SVV exhibited a better ejection fraction (36.19±1.02%,) than GFP controls (26.69±0.75%). NLS-SVV enhanced H3P-positive cardiomyocytes in the border zone to 0.28%, compared to only 0.08% in GFP group, through interacting with Aurora B. Conclusions: We demonstrate the importance of SVV subcellular localization in regulating post-MI cardiac repair and regeneration. We hope that this will open new translational approaches through targeted delivery of SVV.


Arrhythmogenic cardiomyopathy: pathology, genetics, and concepts in pathogenesis.
Arrhythmogenic cardiomyopathy (ACM) is a rare, heritable heart disease characterized by fibro-fatty replacement of the myocardium and a high degree of electric instability. It was first thought to be a congenital disorder, but is now regarded as a dystrophic heart muscle disease that develops over time. There is no curative treatment and current treatment strategies focus on attenuating the symptoms, slowing disease progression, and preventing life-threatening arrhythmias and sudden cardiac death. Identification of mutations in genes encoding desmosomal proteins and in other genes has led to insights into the disease pathogenesis and greatly facilitated identification of family members at risk. The disease phenotype is, however, highly variable and characterized by incomplete penetrance. Although the reasons are still poorly understood, sex, endurance exercise and a gene-dosage effect seem to play a role in these phenomena. The discovery of the genes and mutations implicated in ACM has allowed animal and cellular models to be generated, enabling researchers to start unravelling it's underlying molecular mechanisms. Observations in humans and in animal models suggest that reduced cell-cell adhesion affects gap junction and ion channel remodelling at the intercalated disc, and along with impaired desmosomal function, these can lead to perturbations in signalling cascades like the Wnt/β-catenin and Hippo/YAP pathways. Perturbations of these pathways are also thought to lead to fibro-fatty replacement. A better understanding of the molecular processes may lead to new therapies that target specific pathways involved in ACM.


Simultaneous analysis of six amphetamines and analogues in hair, blood and urine by LC-ESI-MS/MS. Application to the determination of MDMA after low ecstasy intake.
A rapid and sensitive method using LC-MS/MS triple stage quadrupole for the determination of traces of amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy"), 3,4-methylenedioxyethamphetamine (MDEA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in hair, blood and urine has been developed and validated. Chromatography was carried out on an Uptisphere ODB C(18) 5 microm, 2.1 mm x 150 mm column (Interchim, France) with a gradient of acetonitrile and formate 2 mM pH 3.0 buffer. Urine and blood were extracted with Toxitube A (Varian, France). Segmented scalp hair was treated by incubation 15 min at 80 degrees C in NaOH 1M before liquid-liquid extraction with hexane/ethyl acetate (2/1, v/v). The limits of quantification (LOQ) in blood and urine were at 0.1 ng/mL for all analytes. In hair, LOQ was <5 pg/mg for MA, MDMA, MDEA and MBDB, at 14.7 pg/mg for AP and 15.7 pg/mg for MDA. Calibration curves were linear in the range 0.1-50 ng/mL in blood and urine; in the range 5-500 pg/mg for MA, MDMA, MDEA and MBDB, and 20-500 pg/mg for AP and MDA. Inter-day precisions were <13% for all analytes in all matrices. Accuracy was <20% in blood and urine at 1 and 50 ng/mL and <10% in hair at 20 and 250 pg/mg. This method was applied to the determination of MDMA in a forensic case of single administration of ecstasy to a 16-year-old female without her knowledge during a party. She suffered from hyperactivity, sweating and agitation. A first sample of urine was collected a few hours after (T+12h) and tested positive to amphetamines by immunoassay by a clinical laboratory. Blood and urine were sampled for forensic purposes at day 8 (D+8) and scalp hair at day 60 (D+60). No MDMA was detected in blood, but urine and hair were tested positive, respectively at 0.42 ng/mL and at 22 pg/mg in hair only in the segment corresponding to the period of the offence, while no MDA was detectable. This method allows the detection of MDMA up to 8 days in urine after single intake.


Cosmetic powder suspensions in compliant, fingerprintlike contacts.
Cosmetic powders are regularly employed in skin creams and cosmetic formulations to improve performance and enhance skin feel. A previous study investigated the effect of particle concentration and size on the lubricating properties of powder suspensions in smooth, compliant contacts [Timm et al., Tribol. Int. (2011)]. In this paper the tribological properties of cosmetic powder suspensions are investigated in compliant contacts having model fingerprintlike surface topography. Friction coefficients were measured for a series of powder suspensions with varying particle size and concentration in a polydimethylsiloxane (PDMS)/PDMS contact. A commercial tribometer (MTM, PCS Instruments) was employed to measure friction as a function of rubbing time (20 min), under pure sliding (50 mm/s) and low load (0.5 N) conditions. Compared to results using smooth surfaces, it was clear that surface topography has a pronounced affect on the time-dependent tribological behavior of the cosmetic powder suspensions studied. A two-stage friction coefficient versus time curve was observed. By varying the particle size and concentration it was shown that the duration and magnitude of each stage can be controlled.


In vivo sodium release and saltiness perception in solid lipoprotein matrices. 2. Impact of oral parameters.
This study aimed to investigate the relationships between sodium release, saltiness, and oral parameters during the eating of lipoprotein matrices (LPM). Sodium release and saltiness relative to 10 LPM were recorded during normal mastication by five subjects with differing oral parameters (chewing efficiency and salivary flow rate). The LPM samples varied in composition (dry matter, fat, salt, and pH levels) and represented a broad range of hardness. Mastication was recorded using electromyography simultaneously with sensory assessment. Differences in chewing behavior could explain most of the variability in sodium release and saltiness among subjects. Subjects with a higher chewing force and lower salivary flow rate experienced higher levels of sodium release and saltiness. In terms of the LPM, sodium release and saltiness were affected by either chewing behavior or food composition.


Indirect DNA readout on the protein side: coupling between histidine protonation, global structural cooperativity, dynamics, and DNA binding of the human papillomavirus type 16 E2C domain.
DNA sequence recognition by the homodimeric C-terminal domain of the human papillomavirus type 16 E2 protein (E2C) is known to involve both direct readout and DNA-dependent indirect readout mechanisms, while protein-dependent indirect readout has been deduced but not directly observed. We have investigated coupling between specific DNA binding and the dynamics of the unusual E2C fold, using pH as an external variable. Nuclear magnetic resonance and isothermal titration calorimetry show that pH titration of His318 in the complex interface and His288 in the core of the domain is coupled to both binding and the dynamics of the beta-barrel core of E2C, with a tradeoff between dimer stability and function. Specific DNA binding is, in turn, coupled to the slow dynamics and amide hydrogen exchange in the entire beta-barrel, reaching residues far apart from the DNA recognition elements but not affecting the two helices of each monomer. The changes are largest in the dimerization interface, suggesting that the E2C beta-barrel acts as a hinge that regulates the relative position of the DNA recognition helices. In conclusion, the cooperative dynamics of the human papillomavirus type 16 E2C beta-barrel is coupled to sequence recognition in a protein-dependent indirect readout mechanism. The patterns of residue substitution in genital papillomaviruses support the importance of the protonation states of His288 and His318 and suggest that protein-dependent indirect readout and histidine pH titration may regulate DNA binding in the cell.


Mutations disrupting the ordering and topographic mapping of axons in the retinotectal projection of the zebrafish, Danio rerio.
Retinal ganglion cells connect to their target organ, the rectum, in a highly ordered fashion. We performed a large-scale screen for mutations affecting the retinotectal projection of the zebrafish, which resulted in the identification of 114 mutations. 44 of these mutations disturb either the order of RGC axons in the optic nerve and tract, the establishment of a topographic map on the tectum, or the formation of proper termination fields. Mutations in three genes, boxer, dackel and pinscher, disrupt the sorting of axons in the optic tract but do not affect mapping on the tectum. In these mutants, axons from the dorsal retina grow along both the ventral and the dorsal branch of the optic tract. Mutations in two genes, nevermind and who-cares, affect the dorsoventral patterning of the projection. In embryos homozygous for either of these mutations, axons from dorsal retinal ganglion cells terminate ventrally and dorsally in the tectum. In nevermind, the retinotopic order of axons along the optic nerve and tract is changed in a characteristic way as well, while it appears to be unaffected in who-cares. Two mutations in two complementation groups, gnarled and macho, affect the anteroposterior patterning of the projection. In these mutants, nasodorsal axons branch and terminate too soon in the anterior tectum. In 27 mutants belonging to six complementation groups, retinal axons do not form normal termination fields. Some implications for models concerning the formation of topographic projections are discussed.


Escherichia coli-mediated impairment of ureteric contractility is uropathogenic E. coli specific.
Ureters are fundamental for keeping kidneys free from uropathogenic Escherichia coli (UPEC), but we have shown that 2 strains (J96 and 536) can subvert this role and reduce ureteric contractility. To determine whether this is (1) a widespread feature of UPEC, (2) exhibited only by UPEC, and (3) dependent upon type 1 fimbriae, we analyzed strains representing epidemiologically important multilocus sequence types ST131, ST73, and ST95 and non-UPEC E. coli. Contractility and calcium transients in intact rat ureters were compared between strains. Mannose and fim mutants were used to investigate the role of type 1 fimbriae. Non-UPEC had no significant effect on contractility, with a mean decrease after 8 hours of 8.8%, compared with 8.8% in controls. UPEC effects on contractility were strain specific, with decreases from 9.47% to 96.7%. Mannose inhibited the effects of the most potent strains (CFT073 and UTI89) but had variable effects among other UPEC strains. Mutation and complementation studies showed that the effects of the UTI89 cystitis isolate were fimH dependent. We find that (1) non-UPEC do not affect ureteric contractility, (2) impairment of contractility is a common feature of UPEC, and (3) the mechanism varies between strains, but for the most potent UPEC type 1 fimbriae are involved.


User-centered development and testing of a monitoring system that provides feedback regarding physical functioning to elderly people.
To involve elderly people during the development of a mobile interface of a monitoring system that provides feedback to them regarding changes in physical functioning and to test the system in a pilot study. The iterative user-centered development process consisted of the following phases: (1) selection of user representatives; (2) analysis of users and their context; (3) identification of user requirements; (4) development of the interface; and (5) evaluation of the interface in the lab. Subsequently, the monitoring and feedback system was tested in a pilot study by five patients who were recruited via a geriatric outpatient clinic. Participants used a bathroom scale to monitor weight and balance, and a mobile phone to monitor physical activity on a daily basis for six weeks. Personalized feedback was provided via the interface of the mobile phone. Usability was evaluated on a scale from 1 to 7 using a modified version of the Post-Study System Usability Questionnaire (PSSUQ); higher scores indicated better usability. Interviews were conducted to gain insight into the experiences of the participants with the system. The developed interface uses colors, emoticons, and written and/or spoken text messages to provide daily feedback regarding (changes in) weight, balance, and physical activity. The participants rated the usability of the monitoring and feedback system with a mean score of 5.2 (standard deviation 0.90) on the modified PSSUQ. The interviews revealed that most participants liked using the system and appreciated that it signaled changes in their physical functioning. However, usability was negatively influenced by a few technical errors. Involvement of elderly users during the development process resulted in an interface with good usability. However, the technical functioning of the monitoring system needs to be optimized before it can be used to support elderly people in their self-management.


Concerns About Claiming, Postclaim Support, and Return to Work Planning: The Workplace's Impact on Return to Work.
The aim of this study was to determine how injured Australian workers perceived employer emotional (eg, empathy) and instrumental [eg, return-to-work (RTW) planning] support during the RTW process and examine associations between support and RTW. Using data from the 2014 National Return to Work Survey of injured workers with a workers' compensation claim, multinomial regression models examined relationships between support and RTW. Receiving support and developing RTW plans were significantly associated with a greater likelihood of RTW. When controlled for one another in a single model, postclaim support had the strongest association with RTW, with RTW planning also significantly and positively associated with RTW. Provision of both emotional and instrumental support are important employer-led work disability management interventions. Research is required to develop strategies for increasing employer support to lead to improved RTW outcomes for injured workers.


Cabozantinib Inhibits Abiraterone's Upregulation of IGFIR Phosphorylation and Enhances Its Anti-Prostate Cancer Activity.
Abiraterone improves the overall survival of men with metastatic castration-resistant prostate cancer. However, de novo or adaptive resistance to abiraterone limits its activity. Rational combinations of drugs with different mechanisms of action that overcome resistance mechanisms may improve the efficacy of therapy. To that end, we studied the molecular and phenotypic effects of the combination of cabozantinib plus abiraterone. Three prostate cancer cell lines were used to interrogate the in vitro molecular and antiproliferative effects of the single agents and combination of cabozantinib and abiraterone. The in vivo impact of the combination was assessed using the LAPC4-CR xenograft mouse model. In vitro proliferation studies demonstrated single-agent doses between 2 μmol/L and 10 μmol/L for abiraterone and cabozantinib inhibit prostate cancer cell proliferation in a dose-dependent manner, and the anticancer activity of abiraterone is enhanced when combined with cabozantinib. In vivo LAPC4-CR xenograft mouse studies also showed that cabozantinib can improve the antitumor activity of abiraterone. Cabozantinib, a multiple receptor tyrosine kinase inhibitor, enhances the ability of abiraterone to inhibit AR activity in a cell line-dependent manner. In addition, our cell line studies demonstrate abiraterone-stimulated insulin-like growth factor I receptor (IGFIR) phosphorylation with downstream activation of MEK1/2 and ERK1/2, and that this potential adaptive resistance mechanism was inhibited by cabozantinib. Cabozantinib can enhance the efficacy of abiraterone by blocking multiple compensatory survival mechanisms, including IGFIR activation, and supports the assessment of the combination in a clinical trial.


Plasmid associations with residual nuclear structures in Saccharomyces cerevisiae.
Acentric yeast plasmids are mitotically unstable, apparently because they cannot freely diffuse after replicating and therefore are not included in the daughter nucleus. This behavior could result if plasmids remain attached to structural elements of the nucleus after replicating. Since DNA replication is believed to take place on the nuclear matrix, we tested whether there was a correlation between the mitotic stability of a given plasmid and the extent to which it was found associated with residual nuclear structures. Residual nuclei were prepared from yeast nuclei by extraction with either high salt, 2 M NaCl, or low salt, 10 mM lithium diiodosalicylate (LIS). Hybridization analysis was used to estimate the fraction of plasmid molecules remaining after nuclei were extracted. We examined the extent of matrix association of three ARS1 plasmids, Trp1-RI circle (1.45 kb), YRp7 (5.7 kb) and p lambda BAT (45.1 kb) with mitotic loss rates ranging from 3-25%. In addition we examined the matrix binding of the endogenous 2 micron plasmid and the 2 micron-derived YEp13 which is relatively stable in the presence of 2 micron and less stable in cir degree strains. Among the ARS1 plasmids we observed a negative correlation between stability and matrix association, consistent with models in which binding to the nuclear matrix prevents passive segregation of ARS1 plasmid molecules. No such correlation was observed among the 2 micron plasmids. Among all plasmids examined there is a positive correlation between size and matrix association.


Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins.
A new silver stain for electrophoretically separated polypeptides can be rapidly and easily used and can detect as little as 0.01 nanogram of protein per square millimeter. When employed with two-dimensional electrophoresis, it should permit qualitative and quantitative characterization of protein distributions in body fluids and tissues. It has been used to demonstrate regional variations in cerebrospinal fluid proteins.


Opioid growth factor enhances tumor growth inhibition and increases the survival of paclitaxel-treated mice with squamous cell carcinoma of the head and neck.
Paclitaxel is used as a single agent, and in combination with other drugs, as a standard of care in the treatment of squamous cell carcinoma of the head and neck (SCCHN). However, the use of paclitaxel for therapy of SCCHN may be accompanied by serious side effects. Paclitaxel is a known cytotoxic inhibitor of cell proliferation that acts by stabilizing microtubules and inducing apoptosis. Opioid growth factor (OGF), [Met(5)]-enkephalin, is an endogenous peptide that has tonically active inhibitory effects on the growth of SCCHN in vitro and in vivo. OGF action is rapid, reversible, mediated by the nuclear-associated OGF receptor (OGFr), and is not cytotoxic (nor apoptotic related). The present study was designed to examine whether a combination of chemotherapy with paclitaxel and biotherapy with OGF is more effective than either agent alone in inhibiting tumor growth. Moreover, focus was placed on whether there are changes in the side effects known to occur with paclitaxel alone, following this combined therapy. Human SCC-1 cells, derived from a well differentiated SCCHN, were transplanted into athymic mice. The mice were randomized to receive intraperitoneal (i.p.) injections of sterile saline (controls), OGF (10 mg/kg, daily), paclitaxel (8 mg/kg, every other day), or both paclitaxel (8 mg/kg, every other day) and OGF (10 mg/kg, daily) beginning on the day of tumor inoculation. OGF, but not paclitaxel, delayed measurable and visible tumor appearance of mice with SCCHN. Treatment with paclitaxel, but not with other agents, had a marked effect on the body weights. Survival only was reduced in the paclitaxel group, with an average life span of 34.3+/-3.1 days recorded, in comparison to the 50-day survival (date of termination) for all other groups. Beginning after week 4 of tumor inoculation and drug treatment, the tumor weight of the paclitaxel/OGF group was significantly reduced from the control, OGF, and paclitaxel-exposed mice. The OGFr number of the SCCHN tumors was 2.1-fold greater in the animals exposed to OGF or paclitaxel, and elevated 38% in the paclitaxel/OGF group; significant differences from the control group were found for the OGF and paclitaxel groups. These data suggest that combined chemotherapy (i.e., paclitaxel) and biotherapy (OGF) provides a valuable alternative to the standard of care for SCCHN patients.


Multiple forms of short-term plasticity at excitatory synapses in rat medial prefrontal cortex.
Short-term synaptic plasticity, in particular short-term depression and facilitation, strongly influences neuronal activity in cerebral cortical circuits. We investigated short-term plasticity at excitatory synapses onto layer V pyramidal cells in the rat medial prefrontal cortex, a region whose synaptic dynamic properties have not been systematically examined. Using intracellular and extracellular recordings of synaptic responses evoked by stimulation in layers II/III in vitro, we found that short-term depression and short-term facilitation are similar to those described previously in other regions of the cortex. In addition, synapses in the prefrontal cortex prominently express augmentation, a longer lasting form of short-term synaptic enhancement. This consists of a 40-60% enhancement of synaptic transmission which lasts seconds to minutes and which can be induced by stimulus trains of moderate duration and frequency. Synapses onto layer III neurons in the primary visual cortex express substantially less augmentation, indicating that this is a synapse-specific property. Intracellular recordings from connected pairs of layer V pyramidal cells in the prefrontal cortex suggest that augmentation is a property of individual synapses that does not require activation of multiple synaptic inputs or neuromodulatory fibers. We propose that synaptic augmentation could function to enhance the ability of a neuronal circuit to sustain persistent activity after a transient stimulus. This idea is explored using a computer simulation of a simplified recurrent cortical network.


Immunoglobulin M-capture biotin-streptavidin enzyme-linked immunosorbent assay for detection of antibodies to dengue viruses.
A biotin-streptavidin system was adapted to an IgM-capture ELISA for detection of dengue antibodies in human sera. To develop this assay, high titers of antibodies to flavivirus were purified by ion-exchange chromatography (DEAE-cellulose) and labeled with biotin. Heavy chain-specific goat anti-human IgM was first bound to the wells of a polystyrene microtiter plate, followed by binding of IgM in test specimens, and the use of tetravalent dengue antigens (dengue 1-4), biotin-labeled anti-flavivirus IgG, and streptavidin-peroxidase conjugate. The sensitivity and specificity of the IgM-capture biotin-streptavidin ELISA (IgM-BS-ELISA) in acute sera were 83.3% of patients with dengue infection and 95.3% of nondengue-infected cases, respectively. The positive predictive value was 92.4% and the negative predictive value was 89.2%. The efficiency of test was 90.4%. In convalescent sera, the sensitivity and specificity of IgM-BS-ELISA were 100% and 92.6%, respectively. The predictive values of positive and negative results were 90.3% and 100%, respectively. The efficiency of test was 95.6%. The agreement rate of IgM-BS-ELISA and standard hemagglutination inhibition test was good: kappa (kappa) values were 0.79 for acute sera and 0.91 for convalescent sera. The correlation between two methods was quite good, with correlation coefficients (r) of 0.76 for acute sera and 0.85 for convalescent sera (P < 0.001). The results indicate that the IgM-BS-ELISA is highly sensitive, specific, simple to perform, and rapid.


Ifenprodil is a novel type of N-methyl-D-aspartate receptor antagonist: interaction with polyamines.
We have investigated the interactions of polyamines and the N-methyl-D-aspartate (NMDA) receptor antagonist ifenprodil with the binding of [3H]MK801 to the NMDA receptor. Spermine and spermidine but not putrescine substantially increase [3H]MK801 binding to well washed rat brain membranes in the absence or presence of saturating concentrations of glutamate and glycine. Spermine also increased the association and dissociation of [3H]MK801 from its binding site, suggesting that polyamines activate the NMDA receptor in a similar manner to glycine. Ifenprodil inhibited the binding of [3H]MK801 in a biphasic fashion. The high affinity phase of binding (Ki of approximately 15 nM) accounted for 50-60% of total [3H]MK801 binding in the nominal absence of glutamate, glycine, and polyamines or in the presence of 100 microM glutamate. This fraction was reduced to 20% by the addition of 30 microM glycine and could be abolished by the addition of 50 microM spermine. However, ifenprodil apparently did not act by binding to the polyamine recognition site. The low affinity phase (Ki of 20-40 microM) was insensitive to the presence of positive modulators and may represent binding to the Zn2+ regulatory site. Ifenprodil decreased NMDA and glycine-induced Ca2+ influx into cultured rat brain neurons. The potency of ifenprodil suggests that spermine may activate NMDA receptors in vivo. These data indicate that ifenprodil may bind to the NMDA receptor in a state-dependent fashion and preferentially stabilize an inactivated form of the channel.


Electrogenerated chemiluminescence of Ru(bpy)32+ at a black phosphorus quantum dot modified electrode and its sensing application.
Black phosphorus quantum dots (BPQDs) with an average size of 8.2 nm were synthesized through a liquid exfoliation method. The surface morphology and the thickness of the BPQDs were identified by high-resolution transmission microscopy (HRTEM), atomic force microscopy (AFM), and Raman spectroscopy. The electrogenerated chemiluminescence (ECL) behavior of Ru(bpy)32+ was investigated at a BPQD modified glassy carbon electrode under neutral conditions. A strong anodic ECL signal was obtained at the modified electrode in the absence of a coreactant, which is nearly three orders of magnitude larger than that at the bare electrode. Electrochemical results revealed that the oxidation process of Ru(bpy)32+ can be significantly catalyzed by BPQDs, suggesting that BPQDs can act as the coreactant of Ru(bpy)32+ to generate strong light emission. Dopamine could react with the oxidation product of Ru(bpy)32+ and exhibited an apparent inhibiting effect on ECL emission. As a result, it can be sensitively detected in the range of 0.1 nM to 50 nM with a detection limit of 0.022 nM. The present work revealed that BPQDs are a potential ECL platform, and which are promising in the fabrication of a novel ECL biosensor.


Effects of ageing and chronic dietary restriction on the morphology of fast and slow muscles of the rat.
The soleus and extensor digitorum longus muscles of the hindlimb and the flexor digitorum profundus muscle of the forelimb were studied in ad libitum-fed control and age-matched diet-restricted male rats at various ages from weaning to senescence. Growth of individual muscles was accomplished by fibre hypertrophy and not hyperplasia. Between weaning and one year, fibre numbers remained constant in the soleus but fell by 50% in the extensor digitorum longus. Both muscles displayed increasingly oxidative fibre type profiles with advancing age, irrespective of dietary status. This was particularly noticeable in the soleus, which transformed its fibre population from one containing 35% fast fibres at weaning to one with no fast fibres at 91 weeks. In senility, however, the fibre type population again displayed 25% fast fibres. The capillary: fibre ratio and the capillary density were correlated with muscle fibre size in both hindlimb muscles. Although capillarity increased with age, expected differences between fast and slow muscles were probably minimised by the high proportion of FOG fibres in the extensor digitorum longus. Both hindlimb muscles displayed significant increases in the ratio of connective: muscle tissue with increasing age. The soleus invariably contained more connective tissue than the extensor digitorum longus. Dietary restriction reduced the rate of increase, so that the connective tissue content was approximately one half that found in control muscles at one year. Various pathological features associated with old age were delayed considerably in the muscles of the diet-restricted rats. It is concluded that chronic dietary restriction imposed directly after weaning has a dramatic effect on the normal growth and ageing of skeletal muscle.


Robust cue integration: a Bayesian model and evidence from cue-conflict studies with stereoscopic and figure cues to slant.
Most research on depth cue integration has focused on stimulus regimes in which stimuli contain the small cue conflicts that one might expect to normally arise from sensory noise. In these regimes, linear models for cue integration provide a good approximation to system performance. This article focuses on situations in which large cue conflicts can naturally occur in stimuli. We describe a Bayesian model for nonlinear cue integration that makes rational inferences about scenes across the entire range of possible cue conflicts. The model derives from the simple intuition that multiple properties of scenes or causal factors give rise to the image information associated with most cues. To make perceptual inferences about one property of a scene, an ideal observer must necessarily take into account the possible contribution of these other factors to the information provided by a cue. In the context of classical depth cues, large cue conflicts most commonly arise when one or another cue is generated by an object or scene that violates the strongest form of constraint that makes the cue informative. For example, when binocularly viewing a slanted trapezoid, the slant interpretation of the figure derived by assuming that the figure is rectangular may conflict greatly with the slant suggested by stereoscopic disparities. An optimal Bayesian estimator incorporates the possibility that different constraints might apply to objects in the world and robustly integrates cues with large conflicts by effectively switching between different internal models of the prior constraints underlying one or both cues. We performed two experiments to test the predictions of the model when applied to estimating surface slant from binocular disparities and the compression cue (the aspect ratio of figures in an image). The apparent weight that subjects gave to the compression cue decreased smoothly as a function of the conflict between the cues but did not shrink to zero; that is, subjects did not fully veto the compression cue at large cue conflicts. A Bayesian model that assumes a mixed prior distribution of figure shapes in the world, with a large proportion being very regular and a smaller proportion having random shapes, provides a good quantitative fit for subjects' performance. The best fitting model parameters are consistent with the sensory noise to be expected in measurements of figure shape, further supporting the Bayesian model as an account of robust cue integration.


Transcribing pancreas.
For approximately 30-35 years, our insight into some of the fundamental aspects of pancreas development has been based mainly on two independent studies performed in the 1960s by Golosow and Grobstein and Wessells and Cohen. By performing classical embryological experiments, these two reports described the morphogenesis of the pancreas and the epitheliomesenchymal interactions that are required for proper pancreas development. In the 1970s, the groups of LeDourain and associates and Rutter and associates showed, importantly, that despite their similarities with neurons, the pancreatic endocrine cells, like the exocrine and ductual cells, were of an endodermal origin. Then during the 1980s, studies pioneered by Rutter, but also performed by many other groups, were focused on the transcriptional regulation of endocrine and exocrine genes. This eventually lead to the cloning of various transcription factors. By using a genetic approach to study the function of these transcription factors, new insights into pancreas development have now emerged that, on a molecular level, are beginning to explain some of the earlier observations. This review discusses our current knowledge of the mechanisms by which the various pancreatic cell types are generated.


Does surface anodisation of titanium implants change osseointegration and make their extraction from bone any easier?
Titanium implants have a tendency for high bone-implant bonding, and, in comparison to stainless steel implants are more difficult to remove. The current study was carried out to evaluate, i) the release strength of three selected anodized titanium surfaces with increased nanohardness and low roughness, and ii) bone-implant bonding in vivo. These modified surfaces were intended to give improved anchorage while facilitating easier removal of temporary implants. The new surfaces were referenced to a stainless steel implant and a standard titanium implant surface (TiMAX). In a sheep limb model, healing period was 3 months. Bone-implant bonding was evaluated either biomechanically or histologically. The new surface anodized screws demonstrated similar or slightly higher bone-implant-contact (BIC) and torque release forces than the titanium reference. The BIC of the stainless steel implants was significant lower than two of the anodized surfaces (p = 0.04), but differences between stainless steel and all titanium implants in torque release forces were not significant (p = 0.06). The new anodized titanium surfaces showed good bone-implant bonding despite a smooth surface and increased nanohardness. However, they failed to facilitate implant removal at 3 months.


Genomic organization of mouse and human Bruton's agammaglobulinemia tyrosine kinase (Btk) loci.
Btk is a cytoplasmic protein tyrosine kinase (PTK) that has been directly implicated in the pathogenesis of X-linked agammaglobulinaemia (XLA) in humans and X-linked immunodeficiency (Xid) in mice. We have isolated phage and cosmid clones that allowed us to deduce the genomic structure of mouse and human Btk loci. The mouse and human genes are contained within genomic regions that span approximately 43.5 kb and 37.5 kb, respectively. Both loci contain 18 coding exons ranging between 55 and 560 bp in size with introns ranging in size from 164 bp to approximately 9 kb. The 5'-untranslated regions are encoded by single exons located approximately 9 kb upstream of the first coding exon. Exon 18 encodes for the last 23 carboxyl-terminal amino acids and the entire 3'-untranslated region. The location of intron/exon boundaries in the catalytic domains of the mouse and human Btk loci differs from that found in other described sub-families of intracellular PTKs, namely that of Src, Fes/Fer, Csk, and Abl/Arg. This observation is consistent with the classification of Btk together with the recently characterized kinases, Tec and Itk, into a separate sub-family of cytoplasmic PTKs. Putative transcription initiation sites in the mouse and human Btk loci have been determined by using the rapid amplification of cDNA ends assay. Similar to many other PTK specific genes, the putative Btk promoters lack obvious TATAA and CAAAT motifs. Putative initiator elements and potential binding sites for Ets (PEA-3), zeste, and PuF transcription factors are located within the 300 bp which are located upstream of the major transcription start site in both species. These sequences can mediate promoter activity when placed upstream of a promotorless chloramphenicol acetyl transferase reporter gene in an orientation-dependent manner. The present analysis will significantly facilitate the mutational analyses of patients with XLA and the further characterization of the function and regulation of the Btk molecule.