A comparative study of the pollution status of Sakumo II and Muni lagoons in Ghana.
The pollution status of Sakumo II and Muni Lagoons and Mamahuma and Gbagbla Ankonu feeder streams, which feed Sakumo II along the coast of Ghana, were assessed and compared. As expected of saline waters, pH values of both lagoons fell within a slightly neutral to saline range whilst, the feeder streams were moderately saline. Conductivity distribution also showed saline (>5.0 mS cm(-1)) characteristics of the lagoons, while, the feeder streams are brackish (1.5-5.0 mS cm(-1)). DO concentrations in both lagoons indicated a wide variation. Relatively high BOD and nutrients content of Sakumo II and the feeder streams suggested that, Sakumo II and its feeder streams are moderately to grossly polluted. However, Muni Lagoon was characterized by relatively low BOD and nutrients content suggesting relatively low anthropogenic impact. The ionic dominance patterns for Sakumo II and Muni Lagoons were: Na(+) > Mg(2 + ) > Ca(2 + ) and Cl(-) > HCO(3)(-) > SO(4)(2-), Na(+) > Ca(2 + ) > Mg(2 + ) and Cl(-) > HCO(3)(-) > SO(4)(2-), respectively. Using water quality index (WQI), Sakumo II was found to be grossly polluted and requires mitigation measures, while, Muni Lagoon was of fairly good quality and requires monitoring.


Effect of phosphorus concentration on phosphorus removal and biomass.
Membrane bioreactor (MBR) process was employed to study the effect of biological phosphorus removal (bio-P removal) and P-content in treated sludge with increased phosphorus concentration present in the wastewater. Further, the following four test fractions of raw wastewaters was obtained having different P-concentrations viz., run 1: P-20 mg/L, run 2: P-40 mg/L, run 3: P-60 mg/L, run 4: P-80 mg/L. The effective P-removal obtained for these four test fractions were found to be 23.07 mg/L (98.17%), 41.35 mg/L (88.16%), 45.75 mg/L (72.04%) and 55.80 mg/L (66.82%) respectively for run 1, 2, 3 and 4 fractions. Moreover, the similar increase in phosphorous concentration i.e., from 20 to 80 mg/L caused an apparent increase in total solid (TS) values from 7 to 8.3 g TS/L, whereas the total volatile solid (TVS) content remained constant (i.e. 4.5 g TVS/L). These results inferred that the proportion of TVS in the TS decreased from 70 to 55%. Moreover, by increasing the initial P-concentration from 20 to 80 mg/L, the corresponding P-proportion of excess sludge was increased from 2 to 6.2%.


An international comparison of pulmonary rehabilitation: a systematic review.
Despite the strong evidence base, the perception remains that the provision of pulmonary rehabilitation (PR) services are extremely limited. Imbalances in PR delivery give rise to unnecessary health costs and underserviced populations. We conducted a systematic review to characterize the international provision of PR, comparing its structure and delivery across countries, and gaining insight into the availability of PR in relation to geographical prevalence. Electronic databases were searched from inception to September 2013 using the key words "pulmonary rehabilitation" and "survey." Two authors independently reviewed studies and assessed study quality. Data was extracted and double-checked to ensure accuracy. The majority of programs (55-99%) were offered in an outpatient setting with the exception of Ireland, where the majority of programs were offered in the community (65%). Exercise was the primary component across all programs (77-100%), followed by education (74-100%). Physical therapists were the most common member of the PR team (49-100%). Functional walk tests were the most frequently used outcome measure, although the specific test utilized varied across countries. The current availability of PR services ≤1.2% of individuals with COPD. Components provided in PR are similar, irrespective of country, while patient outcome measures demonstrated variation across countries. Recent surveys report the use of community resources for the delivery of PR programs, although the majority are outpatient based. The small number of potential individuals enrolled in PR suggests that an international increase in access and capacity would improve quality of life and reduce healthcare utilization in this population.


Aerobic exercise 2 hours before a dive to 30 msw decreases bubble formation after decompression.
A single bout of aerobic exercise 24 h before a dive significantly reduces the formation of circulating venous gas emboli (VGE) on decompression. The purpose of this investigation was to determine the effect of aerobic exercise 2 h before a dive. There were 16 trained military divers who were compressed to 30 msw (400 kPa) for 30 min breathing air in a dry hyperbaric chamber at rest, then decompressed at a rate of 10 m x min(-1) with a 9-min stop at 3 msw. Each diver performed two dives 3 d apart, one with and one without exercise that consisted of running for 45 min at 60-80% of maximum heart rate (estimated as 220 - age). VGE were graded according to the Spencer scale using a pulsed Doppler detector on the precordium at 30 min (T30) and 60 min (T60) after surfacing. Mean bubble grades at T60 were 1.25 for control dives and 0.44 for dives preceded by exercise, the difference being highly significant. None of the divers showed an increase in venous bubble grade after exercise. Like exercise 24 h ahead, 45 min of running 2 h before a dive decreases bubble formation after diving, suggesting a protective effect of aerobic exercise against DCS. The threshold of exercise intensity and duration necessary to change venous circulating bubbles is unknown. Mechanisms underlying the protective effect of exercise remain unclear. Rather than altering the nitrogen elimination rate, exercise may affect the population of gaseous nuclei from which bubbles form.


Renal involvement and followup of 130 patients with primary Sjögren's syndrome.
To identify the clinical characteristics, pathological changes, and outcome of patients with primary Sjögren's syndrome (pSS). All patients with pSS and renal involvement who were admitted to Ruijin Hospital from April 1993 to December 2006 were included. All the data of clinical features and pathological changes were retrospectively analyzed. Forty-one patients underwent renal biopsies. RESULTS Our study included 130 patients with pSS: 122 women and 8 men. Ages ranged from 16 to 68 years (mean 44.1 +/- 11.52). Ninety-five patients (73.1%) developed renal tubular acidosis (RTA); 91 were found to have distal RTA. Nine patients presented with hypokalemic paralysis. Four patients developed Fanconi syndrome and 3 were proved to have nephrogenic diabetes insipidus. Twenty-seven of 130 patients (20.8%) developed tubular proteinuria and 18/130 (13.8%) presented glomerular involvement. Thirty-five patients (27.7%) developed renal failure (serum creatinine > 115 micromol/l). Most patients (70.8%) had increased serum IgG levels. The incidence of chronic interstitial nephritis was 80.5% among all the biopsy materials. Immunofluorescent staining was negative in most renal tissue. Ninety-six patients were treated with corticosteroids and/or immunosuppressant. Eighteen recovered renal function. Patients with pSS commonly present with renal impairment, mainly from renal tubular dysfunction. The combination of corticosteroids and immunosuppressors significantly improves the renal function of patients with pSS. There is a correlation between hypergammaglobulinemia and distal RTA. The renal acidification capacity for patients with hypergammaglobulinemia. should be monitored.


A clinical prediction rule for nerve-function impairment in leprosy patients.
Nerve-function impairment (NFI) commonly occurs during or after chemotherapy in leprosy and is the key pathological process leading to disability and handicap. We describe the development of a simple clinical prediction rule for estimating the risk of NFI occurrence. New leprosy cases who presented to a centre in Bangladesh were recruited and followed up for 2 years in a field setting. We used multivariable regression analysis by Cox's proportional hazards model to identify predictive variables for NFI. Discriminative ability was measured by a concordance statistic. Internal validity was assessed with bootstrap resampling techniques. 2510 patients were followed up for 2 years, 166 developed NFI. A simple model was developed with leprosy group (either paucibacillary leprosy [PB] or multibacillary leprosy [MB]) and the presence of any nerve-function loss at registration as predictive variables. Patients with PB leprosy and no nerve-function loss had a 1.3% (95% CI 0.8-1.8%) risk of developing NFI within 2 years of registration; patients with PB leprosy and nerve-function loss, or patients with MB leprosy and no nerve-function loss had a 16.0% (12-20%) risk; and patients with MB leprosy with nerve-function loss had a 65% (56-73%) risk. Our prediction rule can be used to plan surveillance of new leprosy patients. Patients at low risk of NFI may need no follow-up beyond their course of chemotherapy (6 months); patients with intermediate risk need a minimum of 1 year of surveillance; and patients with high risk should have at least 2 years of surveillance for new NFI. Current recommendations for surveillance of patients with leprosy (for the duration of chemotherapy only) exclude an important group of patients who are at risk of developing NFI after completion of treatment.


Contemporary thromboprophylaxis of trauma patients.
The traumatically injured patient is at high risk for developing venous thromboembolism. Clinical practice guidelines developed by the American College of Chest Physicians and the Eastern Association for the Surgery of Trauma recognize the importance of initiating thromboprophylaxis, but the guidelines lack specific recommendations regarding the timing and dose of pharmacologic thromboprophylaxis. We review the literature regarding initiation of thromboprophylaxis in different injuries, the use of inferior vena cava filters, laboratory monitoring, dosing regimens, and the use of antiplatelet therapy. Use of pharmacologic thromboprophylaxis with invasive intracranial monitors is not associated with increased bleeding complications. The initiation of low-molecular-weight heparin (LMWH) prophylaxis 48 h postinjury in blunt solid organ injury is not associated with an increase in the rate of failed nonoperative management. Antiplatelet therapy in conjunction with LMWH may help to prevent venous thromboembolism. In the setting of blunt traumatic brain and solid organ injury, initiation of pharmacologic thromboprophylaxis 48 h after injury is not associated with increased bleeding complications. There is no consensus or clear data showing which dosing regimen of LMWH is most effective or whether routine laboratory measurements are beneficial for determining effective thromboprophylaxis.


Tolerance of Superficial Dose to Setup Error With Tomotherapy: Is a Skin Flash Tool Required?
In cancers of the head and neck, gross tumor or areas at risk of microscopic disease often lie close to the skin, while the skin itself may not be at risk. With intensity-modulated radiotherapy, setup errors can lead to underdosage of superficial structures because the collimator will not by default open beyond the skin surface to apply coverage in the air overlying the skin. Thus, small setup errors can move superficial structures out of field for some beams. Some planning systems allow for manually extrapolating fluence for beams tangential to superficial targets. It is unclear whether this problem is significant with tomotherapy. A head and neck phantom was utilized. A 3-mm bolus was used to represent the skin and allow placement of dosimeters at 3 mm depth. Thermoluminescent dosimeters were placed at reproducible points on the skin surface and at 3 mm depth. The phantom was irradiated, with the target volume deep to the thermoluminescent dosimeters receiving a dose of 5 Gy. This process was repeated with the phantom displaced 2.5 mm and again with a displacement of 5 mm. These displacements simulated setup errors that in clinical practice would correspond to bending or twisting of the neck that could not be corrected with rotations or translations. When the phantom was displaced 2.5 mm, the dose measured at 3 mm depth was 99.2% (95.9%-102.5%) of the control. With a 5-mm displacement, the dose at 3 mm only dropped to 91.1% (88.8%-93.4%) of the control. Dose measured at skin surface decreased to a greater degree with such setup error. Dose at superficial depths degraded only slightly with 2.5-mm and even 5-mm displacements. With the tomotherapy system, superficial dose appears to be robust to clinically relevant setup errors. However, if the skin is at risk, bolus should be used to ensure adequate coverage.


Unanticipated Stickiness of α-Pinene.
The adsorption of α-pinene to solid surfaces is an important primary step during the chemical conversion of this common terpene over mesoporous materials, as well as during the formation of atmospheric aerosols. We provide evidence of tight and loose physisorbed states of α-pinene bound on amorphous SiO2 as determined by their adsorption entropy, enthalpy, and binding free energies characterized by computational modeling and vibrational sum frequency generation (SFG) spectroscopy. We find that adsorption is partially (40-60%) irreversible over days at 294-342 K and 1 ATM total pressure of helium, which is supported by molecular dynamics (MD) simulations. The distribution of α-pinene orientation remains invariant with temperature and partial pressure of α-pinene. Using the Redlich-Peterson adsorption model in conjunction with a van't Hoff analysis of adsorption isotherms recorded for up to 2.6 Torr α-pinene in 1 ATM total pressure of helium, we obtain ΔS°ads, ΔH°ads, and ΔG°ads values of -57 (±7) J mol-1 K-1, -39 (±2) kJ mol-1, and -22 (±5) kJ mol-1, respectively, associated with the reversibly bound population of α-pinene. These values are in good agreement with density functional theory (DFT)-corrected force field calculations based on configurational sampling from MD simulations. Our findings are expected to have direct implications on the conversion of terpenes by silica-based catalysts and for the synthesis of secondary organic aerosol (SOA) in atmospheric chambers and flow tubes.


A systematic review and meta-analysis of the nutrient content of preterm and term breast milk.
Breast milk nutrient content varies with prematurity and postnatal age. Our aims were to conduct a meta-analysis of preterm and term breast milk nutrient content (energy, protein, lactose, oligosaccharides, fat, calcium, and phosphorus); and to assess the influence of gestational and postnatal age. Additionally we assessed for differences by laboratory methods for: energy (measured vs. calculated estimates) and protein (true protein measurement vs. the total nitrogen estimates). Systematic review results were summarized graphically to illustrate the changes in composition over time for term and preterm milk. Since breast milk fat content varies within feeds and diurnally, to obtain accurate estimates we limited the meta-analyses for fat and energy to 24-hour breast milk collections. Forty-one studies met the inclusion criteria: 26 (843 mothers) preterm studies and 30 (2299 mothers) term studies of breast milk composition. Preterm milk was higher in true protein than term milk, with differences up to 35% (0.7 g/dL) in colostrum, however, after postnatal day 3, most of the differences in true protein between preterm and term milk were within 0.2 g/dL, and the week 10-12 estimates suggested that term milk may be the same as preterm milk by that age. Colostrum was higher than mature milk for protein, and lower than mature milk for energy, fat and lactose for both preterm and term milk. Breast milk composition was relatively stable between 2 and 12 weeks. With milk maturation, there was a narrowing of the protein variance. Energy estimates differed whether measured or calculated, from -9 to 13%; true protein measurement vs. the total nitrogen estimates differed by 1 to 37%. Although breast milk is highly variable between individuals, postnatal age and gestational stage (preterm versus term) were found to be important predictors of breast milk content. Energy content of breast milk calculated from the macronutrients provides poor estimates of measured energy, and protein estimated from the nitrogen over-estimates the protein milk content. When breast milk energy, macronutrient and mineral content cannot be directly measured the average values from these meta-analyses may provide useful estimates of mother's milk energy and nutrient content.


Measurement of Oxygen Status in Arabidopsis Leaves Undergoing the Hypersensitive Response During Pseudomonas Infection.
When plants are infected with pathogens they defend themselves via various processes. Once such process is the development of the hypersensitive response (HR), a kind of programmed cell death (PCD), in which localized cell death takes place in order to prevent pathogen spread to other part of tissue. The Arabidopsis and Pseudomonas syringae system is one of the best known examples to study the HR. Here we used the VisiSens™ oxygen-imaging system to investigate oxygen distributions in Arabidopsis leaves infected with an avirulent strain of P. syringae and undergoing the hypersensitive response. Using this method we observed a change in oxygen status at 6 h post-infection and a drop in oxygen levels at 24 h after infection.


Characterization of Visual Function, Interocular Variability and Progression Using Static Perimetry-Derived Metrics in RPGR-Associated Retinopathy.
To characterize bilateral visual function, interocular variability and progression by using static perimetry-derived volumetric and pointwise metrics in subjects with retinitis pigmentosa associated with mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. This was a prospective longitudinal observational study of 47 genetically confirmed subjects. Visual function was assessed with ETDRS and Pelli-Robson charts; and Octopus 900 static perimetry using a customized, radially oriented 185-point grid. Three-dimensional hill-of-vision topographic models were produced and interrogated with the Visual Field Modeling and Analysis software to obtain three volumetric metrics: VTotal, V30, and V5. These were analyzed together with Octopus mean sensitivity values. Interocular differences were assessed with the Bland-Altman method. Metric-specific exponential decline rates were calculated. Baseline symmetry was demonstrated by relative interocular difference values of 1% for VTotal and 8% with V30. Degree of symmetry varied between subjects and was quantified with the subject percentage interocular difference (SPID). SPID was 16% for VTotal and 17% for V30. Interocular symmetry in progression was greatest when quantified by VTotal and V30, with 73% and 64% of subjects possessing interocular rate differences smaller in magnitude than respective annual progression rates. Functional decline was evident with increasing age. An overall annual exponential decline of 6% was evident with both VTotal and V30. In general, good interocular symmetry exists; however, there was both variation between subjects and with the use of various metrics. Our findings will guide patient selection and design of RPGR treatment trials, and provide clinicians with specific prognostic information to offer patients affected by this condition.


Survival of developing motor neurons mediated by Rho GTPase signaling pathway through Rho-kinase.
A variety of neurons generated during embryonic development survive or undergo programmed cell death (PCD) at later developmental stages. Survival or death of developing neurons is generally considered to depend on trophic support from various target tissues. The small GTPase Rho regulates diverse cellular processes such as cell morphology, cell adhesion, cell motility, and apoptosis. Rho-dependent serine-threonine protein kinase (Rho-kinase-ROK-ROCK), one of the effector proteins, transmits signals for some Rho-mediated processes. Here, we report the in vivo role of the Rho signaling pathway through Rho-kinase during development of motor neurons (MNs) in the spinal cord. We performed conditional expression of a dominant-negative form for RhoA (RhoA DN) or for Rho-kinase (Rho-K DN) in transgenic mice by using the Cre-loxP system to suppress the activity of these signaling molecules in developing MNs. Expression of RhoA DN reduced the number of MNs in the spinal cord because of increased apoptosis while preserving the gross patterning of motor axons. Expression of Rho-K DN produced developmental defects similar to those observed in RhoA DN expression. In addition, analysis of transgenic mice expressing Rho-K DN showed that the increased apoptosis of MNs was induced at the early embryonic stages before the initiation of PCD, and that MN death at the late embryonic stages corresponding to the period of PCD was moderately enhanced in the transgenic mice. These findings indicate that the Rho signaling pathway, primarily through Rho-kinase, plays a crucial role in survival of spinal MNs during embryogenesis, particularly at the early developmental stages.


First dose ChAdOx1 and BNT162b2 COVID-19 vaccinations and cerebral venous sinus thrombosis: A pooled self-controlled case series study of 11.6 million individuals in England, Scotland, and Wales.
Several countries restricted the administration of ChAdOx1 to older age groups in 2021 over safety concerns following case reports and observed versus expected analyses suggesting a possible association with cerebral venous sinus thrombosis (CVST). Large datasets are required to precisely estimate the association between Coronavirus Disease 2019 (COVID-19) vaccination and CVST due to the extreme rarity of this event. We aimed to accomplish this by combining national data from England, Scotland, and Wales. We created data platforms consisting of linked primary care, secondary care, mortality, and virological testing data in each of England, Scotland, and Wales, with a combined cohort of 11,637,157 people and 6,808,293 person years of follow-up. The cohort start date was December 8, 2020, and the end date was June 30, 2021. The outcome measure we examined was incident CVST events recorded in either primary or secondary care records. We carried out a self-controlled case series (SCCS) analysis of this outcome following first dose vaccination with ChAdOx1 and BNT162b2. The observation period consisted of an initial 90-day reference period, followed by a 2-week prerisk period directly prior to vaccination, and a 4-week risk period following vaccination. Counts of CVST cases from each country were tallied, then expanded into a full dataset with 1 row for each individual and observation time period. There was a combined total of 201 incident CVST events in the cohorts (29.5 per million person years). There were 81 CVST events in the observation period among those who a received first dose of ChAdOx1 (approximately 16.34 per million doses) and 40 for those who received a first dose of BNT162b2 (approximately 12.60 per million doses). We fitted conditional Poisson models to estimate incidence rate ratios (IRRs). Vaccination with ChAdOx1 was associated with an elevated risk of incident CVST events in the 28 days following vaccination, IRR = 1.93 (95% confidence interval (CI) 1.20 to 3.11). We did not find an association between BNT162b2 and CVST in the 28 days following vaccination, IRR = 0.78 (95% CI 0.34 to 1.77). Our study had some limitations. The SCCS study design implicitly controls for variables that are constant over the observation period, but also assumes that outcome events are independent of exposure. This assumption may not be satisfied in the case of CVST, firstly because it is a serious adverse event, and secondly because the vaccination programme in the United Kingdom prioritised the clinically extremely vulnerable and those with underlying health conditions, which may have caused a selection effect for individuals more prone to CVST. Although we pooled data from several large datasets, there was still a low number of events, which may have caused imprecision in our estimates. In this study, we observed a small elevated risk of CVST events following vaccination with ChAdOx1, but not BNT162b2. Our analysis pooled information from large datasets from England, Scotland, and Wales. This evidence may be useful in risk-benefit analyses of vaccine policies and in providing quantification of risks associated with vaccination to the general public.


Expression of transformation-associated protease(s) that degrade fibronectin at cell contact sites.
Virus-transformed fibroblasts show an increased production of proteases as well as loss of extracellular adhesive proteins. To determine whether these transformation-associated events are related, we investigated the capacity of Rous sarcoma virus-transformed cells (embryonic chick fibroblasts and mouse BALB/c 3T3) to degrade fibronectin by using a novel cross-linked protein substratum: fluorescence-labeled or radiolabeled fibronectin covalently linked to the surface of a fixed gelatin film. In serum-containing medium, the coupled fibronectin was not released when incubated without cells, and only a small amount was released when incubated with nontransformed cells. However, when transformed cells were seeded on the radiolabeled fibronectin-coupled substratum, there was a threefold increase in the time-dependent release of radioactivity into the medium. The released material was characterized as peptides with molecular sizes of less than 30,000 daltons. Correspondingly, growth of transformed cells on the rhodamine-fibronectin substratum resulted in the appearance of discrete negative fluorescent spots beneath the cells and along their migratory paths, whereas a uniform fluorescent carpet was detected with nontransformed cells. The release of radioactivity was partially inhibited by protease inhibitors, including alpha 2-macroglobulin, leupeptin, and benzamidine, but the negative fluorescent spots appeared unaffected by any of these inhibitors. However, both the release of radiolabeled peptides and the appearance of fluorescence-negative spots were inhibited by 1,10-phenanthroline at concentrations that did not affect cellular attachment and protein synthesis, thus supporting a role for proteases in localized degradation of fibronectin substratum. These fluorescence-negative spots coincided with sites of fibronectin disappearance as judged by indirect labeling with antibodies to cellular fibronectin. In addition, immunofluorescent analyses showed a correlation between vinculin localization and the negative fibronectin spots found under transformed cells, indicating that degradation occurs at cell substratum contact sites. These results can be correlated with other transformation-associated phenotypic changes, and are discussed in terms of the invasion of tumor cells into the extracellular matrix.


Father mental health during the early parenting period: results of an Australian population based longitudinal study.
The primary objective of this study was to report on the occurrence of mental health difficulties for a large national sample of Australian fathers of children aged 0-5 years (n = 3,471). Secondary objectives were to compare fathers' mental health against normative data for the general male adult population, and to examine the course of mental health problems for fathers across the early childhood period. Secondary analysis of data from the infant cohort of the Longitudinal Study of Australian Children at three waves when children were 0-12 months, 2-3 and 4-5 years. Comparative data on the prevalence of psychological distress in the Australian adult male population sourced from the National Survey of Mental Health and Wellbeing. Approximately nine per cent of fathers reported symptomatic or clinical psychological distress at each wave, as measured by the Kessler-6. Approximately 30 % reporting distress at wave 1 continued to report distress at a similar or worse level across waves 2 and 3. Fathers not living with their children also had high rates of distress (14 % at wave 1 and 10 % at wave 2). Finally, fathers in the present study had 1.38 increased odds (95 % CI 1.12-1.69) for psychological distress compared with the Australian adult male population. Fathers are at risk of experiencing postnatal mental health difficulties, which may persist across the early childhood period for some fathers. The results suggest routine assessment of fathers' wellbeing should be undertaken in the postnatal period with mental health interventions and support provided across the early childhood period.


Computed tomography myelographic findings in dogs with cervical spondylomyelopathy.
Computed tomography (CT) myelography is used occasionally in the diagnosis of cervical spondylomyelopathy, but the type of lesion found in large- versus giant-breed dogs using this modality has not been characterized. Our purpose was to report the frequency of compressive lesions in large- and giant-breed dogs with cervical spondylomyelopathy and imaged using CT myelography. Fifty-eight dogs were retrospectively studied, 23 large-breed and 35 giant-breed dogs. Multiple sites of compression were found in 12 large-breed dogs (52.2%) compared to 30 (85.8%) giant-breed dogs. The main site of compression was at C5-6 and C6-7 in both large-breed (91.3%) and giant-breed (72.4%) dogs. The main cause and direction of compression was disc-associated and ventral in 19 (82.6%) of the large-breed dogs while osseous changes were the primary cause of compression in 27 (77.2%) of the giant-breed dogs, with most compressions being lateral (51.4%), followed by dorsolateral (14.2%). Osseous compression was observed at C7-T1 in eight giant-breed dogs (22.8%), and at T1-T2 or T2 only in five dogs (14.3%). Four of 23 large-breed dogs (17.4%), and seven (20%) of 35 giant-breed dogs had spinal cord atrophy. Therefore, giant-breed dogs often have multiple compressions, usually caused by osseous changes causing lateralized compressions. In large-breed dogs most compressions are disc-associated and located ventrally. Considering the number of giant-breed dogs with compressions at C7-T1, T1-2, and T2, it is important to include the cranial thoracic region when imaging dogs suspected of having cervical spondylomyelopathy.


Solid-state NMR covariance of homonuclear correlation spectra.
Direct covariance NMR spectroscopy, which does not involve a Fourier transformation along the indirect dimension, is demonstrated to obtain homonuclear correlation two-dimensional (2D) spectra in the solid state. In contrast to the usual 2D Fourier transform (2D-FT) NMR, in a 2D covariance (2D-Cov) spectrum the spectral resolution in the indirect dimension is determined by the resolution along the detection dimension, thereby largely reducing the time-consuming indirect sampling requirement. The covariance method does not need any separate phase correction or apodization along the indirect dimension because it uses those applied in the detection dimension. We compare in detail the specifications obtained with 2D-FT and 2D-Cov, for narrow and broad resonances. The efficiency of the covariance data treatment is demonstrated in organic and inorganic samples that are both well crystallized and amorphous, for spin -1/2 nuclei with 13C, 29Si, and 31P through-space or through-bond homonuclear 2D correlation spectra. In all cases, the experimental time has been reduced by at least a factor of 10, without any loss of resolution and signal to noise ratio, with respect to what is necessary with the 2D-FT NMR. According to this method, we have been able to study the silicate network of glasses by 2D NMR within reasonable experimental time despite the very long relaxation time of the 29Si nucleus. The main limitation of the 2D-Cov data treatment is related to the introduction of autocorrelated peaks onto the diagonal, which does not represent any actual connectivity.


The effect of external calcium and magnesium depletion on single nerve fibers.
The three types of motor axons found in the walking legs of the lobster were shown to respond differently upon exposure to calcium-free solutions. While all fiber types became more excitable initially in calcium-free solutions, only openers became spontaneously active. Fast closers showed the least reduction in rheobase value upon calcium depletion. After 5 minutes in calcium-free solution all fibers showed a rise in rheobase value, and more rapid accommodation. A natural period for spontaneous firing of opener fibers was disclosed. Following such a spontaneous discharge, low amplitude rhythmical potentials were recorded. These small potentials had the same period as the spontaneous spikes. The role of calcium ion in the excitable process was discussed. Magnesium ion was shown to act synergistically with calcium ion. All fiber types became spontaneously active in solutions deprived of both calcium and magnesium. Subsequent hypoexcitability was more pronounced in calcium- and magnesium-depleted solutions than it was in only calcium-depleted solutions.


Involvement of leucine residues at positions 107, 112, and 115 in a leucine-rich repeat motif of human Toll-like receptor 2 in the recognition of diacylated lipoproteins and lipopeptides and Staphylococcus aureus peptidoglycans.
S-(2,3-bispalmitoyloxypropyl)Cys-Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe (FSL-1) derived from Mycoplasma salivarium stimulated NF-kappaB reporter activity in human embryonic kidney 293 (HEK293) cells transfected with Toll-like receptor 2 (TLR2) or cotransfected with TLR2 and TLR6, but not in HEK293 cells transfected with TLR6, in a dose-dependent manner. The activity was significantly higher in HEK293 cells transfected with both TLR2 and TLR6 than in HEK293 cells transfected with only TLR2. The deletion mutant TLR2(DeltaS40-I64) (a TLR2 mutant with a deletion of the region of Ser(40) to Ile(64)) failed to activate NF-kappaB in response to FSL-1. The deletion mutant TLR2(DeltaC30-S39) induced NF-kappaB reporter activity, but the level of activity was significantly reduced compared with that induced by wild-type TLR2. A TLR2 point mutant with a substitution of Glu(178) to Ala (TLR2(E178A)), TLR2(E180A), TLR2(E190A), and TLR2(L132E) induced NF-kappaB activation when stimulated with FSL-1, M. salivarium lipoproteins, and Staphylococcus aureus peptidoglycans, but TLR2(L107E), TLR2(L112E) (a TLR2 point mutant with a substitution of Leu(112) to Glu), and TLR2(L115E) failed to induce NF-kappaB activation, suggesting that these residues are essential for their signaling. Flow cytometric analysis demonstrated that TLR2(L115E), TLR2(L112E), and TLR2(DeltaS40-I64) were expressed on the cell surface of the transfectants as wild-type TLR2 and TLR2(E190A) were. In addition, these mutants, except for TLR2(E180A), functioned as dominant negative form of TLR2. This study strongly suggested that the extracellular region of Ser(40)-Ile(64) and leucine residues at positions 107, 112, and 115 in a leucine-rich repeat motif of TLR2 are involved in the recognition of mycoplasmal diacylated lipoproteins and lipopeptides and in the recognition of S. aureus peptidoglycans.


Candidate tumor suppressor BTG3 maintains genomic stability by promoting Lys63-linked ubiquitination and activation of the checkpoint kinase CHK1.
B-cell translocation gene 3 (BTG3) is a member of the antiproliferative BTG/ Transducer of ErbB2 gene family and is induced by genotoxic stress in a p53- and Checkpoint kinase 1 (CHK1)-dependent manner. Down-regulation of BTG3 has been observed in human cancers, suggesting that it plays an important role in tumor suppression, although the underlying mechanisms are unclear. Here, we report that BTG3 interacts with CHK1, a key effector kinase in the cell cycle checkpoint response, and regulates its phosphorylation and activation. Upon interaction, BTG3 mediates K63-linked ubiquitination of CHK1 at Lys132 through the cullin-RING ligase 4(Cdt2) E3 complex, thus facilitating CHK1 chromatin association. We show that BTG3-depleted cells phenocopy those CHK1-deficient cells, exhibiting increased cell death after replication block and impaired chromosome alignment and segregation. These defects could be corrected by wild-type BTG3 but not by a mutant impaired in CHK1 interaction. We propose that BTG3-dependent CHK1 ubiquitination contributes to its chromatin localization and activation and that a defect in this regulation may increase genome instability and promote tumorigenesis.


In situ observation of the internal structure and composition of biomineralized Emiliania huxleyi calcite by solid-state NMR Spectroscopy.
Biomineralization, particularly the formation of calcium carbonate structures by organisms under ambient conditions, is of vast fundamental and applied interest. Organisms finely control all aspects of the formation of the biomaterials: composition, polymorph, morphology, and macroscopic properties. While in situ molecular-level characterization of the resulting biominerals is a formidable task, solid-state magic angle spinning NMR is one of the most powerful analytical techniques for this purpose. It is employed in this study to elucidate the structure and composition of biogenic calcite formed by Emiliania huxleyi, a unicellular alga distinguished by its exquisitely sculptured calcite cell coverings known as coccoliths. Strain 371 (CCMP) was grown and harvested from (15)N- and (13)C-enriched growth medium, with biosynthetic labeling to enhance the sensitivity of the NMR measurements. Crystalline and interfacial calcite environments were selectively probed using direct and indirect (cross-polarized) (13)C excitation, respectively. Different crystalline environments, in particular structural defect sites at concentrations of up to 1.4% with P and N moieties incorporated, were identified using (13)C rotational-echo double-resonance (REDOR) NMR. REDOR-derived geometrical constraints show that the P and N atoms at the defect sites are 3.2 and 2.3 (+/-0.2) A apart from a crystalline carbon carbonate. The phosphorus and nitrogen moieties within the biogenic calcite are identified as small, non-protonated moieties, attributed to inorganic ions such as PO4(3-) and NO3(-). The carbonates adjacent to these defects are chemically indistinguishable from bulk crystalline carbonates, yet their immediate environments experience reduced rigidity, as reflected by substantial T1((13)CO3(2-)) shortening. Interfacial carbonates, on the other hand, reside in structurally/chemically perturbed environments, as reflected by heterogeneous line broadening. This study is the first to directly unravel evidence on the incorporation of P/N moieties as structural defects within E. huxleyi biogenic calcite, and on the state of the adjacent crystalline carbonates.


Comparing caudal and intravenous ketamine for supplementation of analgesia after Salter innominate osteotomy.
Previous studies claim that caudal administration of ketamine causes effective analgesia. The aim of this study was to assess the clinical effectiveness of ketamine after caudal or intravascular administration in pediatric patients that underwent orthopedic surgery to distinguish between local and systemic analgesia. After the induction of general anesthesia, 36 patients, aged 18 months to 10 years, assigned to undergo orthopedic surgery, received a caudal injection of bupivacaine and were randomly blinded into two groups: one group received 1 mg/kg S(+)-ketamine as the caudal group and the other group received 1 mg/kg S(+)-ketamine as the intravascular group. Postsurgical measurements included the effectiveness of postsurgical analgesia, which was assessed by using the observational pain scale (OPS), duration of analgesia, sedation score, and hemodynamic and respiratory monitoring. The mean time to first analgesia was clearly longer in the caudal ketamine group (13.35 h) than in the intravenous ketamine (9.93 h) group (P < 0.01). During the 24-h observation time, fewer children asked for additional analgesic drugs in the caudal group (8 of 18, 44.4 %) than in the intravenous group (12 of 18, 66.6 %; P = 0.01). The times to first micturation and spontaneous leg movements and the incidence of nausea and vomiting were similar in the two groups. The OPS and sedation scores after operation showed no obvious differences between the groups at any time. Although caudal ketamine provides good postsurgical analgesia due to its potential neurotoxicity and only small clinical differences with intravenous ketamine, the administration of intravenous ketamine might be a reasonable option to potentially extend the postsurgical analgesic effect of the caudal administration of local anesthetics in children undergoing Salter osteotomy.


Stroke Severity Affects Timing: Time From Stroke Code Activation to Initial Imaging is Longer in Patients With Milder Strokes.
Optimizing the time it takes to get a potential stroke patient to imaging is essential in a rapid stroke response. At our hospital, door-to-imaging time is comprised of 2 time periods: the time before a stroke is recognized, followed by the period after the stroke code is called during which the stroke team assesses and brings the patient to the computed tomography scanner. To control for delays due to triage, we isolated the time period after a potential stroke has been recognized, as few studies have examined the biases of stroke code responders. This "code-to-imaging time" (CIT) encompassed the time from stroke code activation to initial imaging, and we hypothesized that perception of stroke severity would affect how quickly stroke code responders act. In consecutively admitted ischemic stroke patients at The Mount Sinai Hospital emergency department, we tested associations between National Institutes of Health Stroke Scale scores (NIHSS), continuously and at different cutoffs, and CIT using spline regression, t tests for univariate analysis, and multivariable linear regression adjusting for age, sex, and race/ethnicity. In our study population, mean CIT was 26 minutes, and mean presentation NIHSS was 8. In univariate and multivariate analyses comparing CIT between mild and severe strokes, stroke scale scores <4 were associated with longer response times. Milder strokes are associated with a longer CIT with a threshold effect at a NIHSS of 4.


Monomer-dimer equilibrium constants of RNA in the dimer initiation site of human immunodeficiency virus type 1.
The genome of the human immunodeficiency virus (HIV) exists as a dimer of two identical RNA molecules hydrogen bonded to each other near their 5' ends. The dimer, known to be important for viral infectivity, is formed by two monomers interacting through a stem-loop structure called the dimer initiation site (DIS). An initially formed intermediate, the "kissing" dimer, is unstable and rearranges to the stable, duplex form. In this report we use nondenaturing polyacrylamide gel electrophoresis to measure the monomer-dimer equilibrium constant of three RNA sequences, 41-, 27-, and 19-mers, located in the DIS of the MAL isolate of HIV-1. Experiments in which the RNA was equilibrated at various temperatures before electrophoresis revealed that interconversion is rapid for all the sequences, so that they reach equilibrium in the loading well of the gel at 5 degrees C before they enter the gel proper. However, interconversion kinetics in the gel are slow, so autoradiographic spot intensities can be used to measure the amounts of monomer and dimer present when the sample entered the gel. After correction for the amount of RNA added with the radiolabel and dilution of samples in the loading well of the gel, dimerization equilibrium constants were calculated from spot intensities. The calculated values of the dimerization constant K at 5 degrees C were approximately 10(5), approximately 10(6), and approximately 10(8) M(-1) for the 41-, 27-, and 19-mers, respectively, in solutions of ionic strength, I, of about 100 mM. The decrease in K by three orders of magnitude between the 19-mer and 41-mer is due in part to the change in rotational entropy of rodlike molecules on dimerization and in part to the increased conformational entropy of the monomers. As expected, increased ionic strength increases the dimerization constant for all three RNAs. For the 41-mer, however, K has a maximum value at I approximately 140 mM. The origin of the decrease in K for higher I is unknown but it may be due to formation of species (perhaps higher order oligomers) that do not enter the gel. The 41-mer exists in two dimeric forms assigned to the kissing and duplex dimers. The ratio of kissing to duplex form at 5 degrees C is 0.48 +/- 0.22 at I = 113 mM and 0.91 +/- 0.35 at I = 183 mM. The observed decrease in K with RNA length suggests that the dimerization constant of the packaging region of HIV-1 is small, < approximately 10(5) M(-1), implying that the nucleocapsid protein is important in promoting dimerization in the capsid of the virus.


Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis.
Elementary bodies (EB) of Chlamydia trachomatis serotypes C, E, and L2 were extrinsically radioiodinated, and whole-cell lysates of these serotypes were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Autoradiography of the polypeptide profiles identified a major surface protein with an apparent subunit molecular weight of 39,500 that was common to each C. trachomatis serotype. The abilities of nonionic (Triton X-100), dipolar ionic (Zwittergent TM-314), mild (sodium deoxycholate and sodium N-lauroyl sarcosine), and strongly anionic (SDS) detergents to extract this protein from intact EB of the L2 serotype were investigated by SDS-PAGE analysis of the soluble and insoluble fractions obtained after each detergent treatment. Only SDS readily extracted this protein from intact EB. Sarkosyl treatment selectively solubilized the majority of other EB proteins, leaving the 39,500-dalton protein associated with the Sarkosyl-insoluble fraction. Ultrastructural studies of the Sarkosyl-insoluble EB pellet showed it to consist of empty EB particles possessing an apparently intact outer membrane. No structural evidence for a peptidoglycan-like cell wall was found. Morphologically these chlamydial outer membrane complexes (COMC) resembled intact chlamydial EB outer membranes. The 39,500-dalton outer membrane protein was quantitatively extracted from COMC by treating them with 2% SDS at 60 degrees C. This protein accounted for 61% of the total COMC-associated protein, and its extraction resulted in a concomitant loss of the COMC membrane structure and morphology. The soluble extract obtained from SDS-treated COMC was adsorbed to a hydroxylapatite column and eluted with a linear sodium phosphate gradient. The 39,500-dalton protein was eluted from the column as a single peak at a phosphate concentration of approximately 0.3 M. The eluted protein was nearly homogeneous by SDS-PAGE and appeared free of contaminating carbohydrate, glycolipid, and nucleic acid. Hyperimmune mouse antiserum prepared against the 39,500-dalton protein from serotype L2 reacted with C. trachomatis serotypes Ba, E, D, K, L1, L2, and L3 by indirect immunofluorescence with EB but failed to react with serotypes A, B, C, F, G, H, I, and J, with the C. trachomatis mouse pneumonitis strain, or with the C. psittaci feline pneumonitis, guinea pig inclusion conjunctivitis, or 6BC strains. Thus, the 39,500-dalton major outer membrane protein is a serogroup antigen of C. trachomatis organisms.


Metallicity retained by covalent functionalization of graphene with phenyl groups.
To resolve the controversy over the functionalization effect on conductivity, we systematically investigate the structural and electronic properties of graphene covalently functionalized with phenyl groups. Using first-principles calculations combined with the model Hamiltonian analysis, we find that the structural stability, electronic and transport properties of the functionalized graphene are strongly dependent on the adsorption site of the phenyl groups. In detail, double-side functionalized graphene is energetically more favorable than single-side functionalized graphene, and more importantly, they exhibit an exotic non-magnetic metallic state and a magnetic semiconducting state, respectively. For covalently double-side functionalized graphene, two bands contributed by π electrons of graphene cross at the Fermi level with the preserved electron-hole symmetry, and the Fermi velocity of carriers could be flexibly tuned by changing the coverage of the phenyl groups. These results provide an insight into the experimental observation [ACS Nano 2011, 5, 7945], interpreting the origin of the increase in the conductivity of graphene covalently functionalized with phenyl groups. Our work reveals the great potential of these materials in future nanoelectronics or sensors by controlling the attachment of phenyl groups.


Validity of an ecometric neighborhood physical disorder measure constructed by virtual street audit.
Neighborhood physical disorder is thought to affect mental and physical health, but it has been difficult to measure objectively and reliably across large geographical areas or multiple locales. Virtual street audits are a novel method for assessing neighborhood characteristics. We evaluated the ecometric properties of a neighborhood physical disorder measure constructed from virtual street audit data. Eleven trained auditors assessed 9 previously validated items developed to capture physical disorder (e.g., litter, graffiti, and abandoned buildings) on 1,826 block faces using Google Street View imagery (Google, Inc., Mountain View, California) dating from 2007-2011 in 4 US cities (San Jose, California; Detroit, Michigan; New York, New York; and Philadelphia, Pennsylvania). We constructed a 2-parameter item response theory scale to estimate latent levels of disorder on each block face and defined a function using kriging to estimate physical disorder levels, with confidence estimates, for any point in each city. The internal consistency reliability of the resulting scale was 0.93. The final measure of disorder was positively correlated with US Census data on unemployment and housing vacancy and negatively correlated with data on owner-occupied housing. These results suggest that neighborhood physical disorder can be measured reliably and validly using virtual audits, facilitating research on possible associations between physical disorder and health.


Microbial colonization of the peri-implant environment and its relevance to long-term success of osseointegrated implants.
This article provides a critical examination of the literature on the colonization of the peri-implant environment and its impact on clinically significant infection, in the context of a comparison with the microbial etiology of periodontitis. Previous journal article bibliographies and MEDLINE searches provided comprehensive access to current literature on this topic. References that provided significant or novel advancements in knowledge were included in this review. Since it was prepared for a workshop that would examine this issue critically, this review offers provocative commentary. There are many similarities in peri-implant and periodontal microbiology, both in health and in infection leading to progressive deterioration. Periodontal pockets serve as reservoirs for the colonization of implants by periodontal pathogens in partially edentulous patients. Implant design and material composition affect the colonization of implant surfaces. Infections around implants can be treated by similar strategies used for periodontitis. Investigators are beginning to adopt research designs used in periodontology to study the pathogenesis and treatment of peri-implant infections. Research in this area is impeded by the lack of surrogate variables for crestal bone resorption. Some implants fail as a result of infection, but the frequency of failure by infection or any other cause is unknown. This paper suggests areas where much more additional research is needed, including critical consideration of strategies that have led to the past few decades' advancements in the pathogenesis of periodontitis.


Polyoxyethylene as the stationary phase in ion chromatography.
The present article reviews the use of polyethylene glycol (PEG) or polyoxyethylene (POE) as the stationary phase for the separation of inorganic anions in ion chromatography and discusses about the retention mechanisms involved in the separation of anions on the novel stationary phases. PEG permanently coated on a hydrophobic stationary phase retained anions in the partition mode and allowed us to use high-concentration eluents because the retention of anions increased with increasing eluent concentration for most of the eluents. This situation was convenient to determine trace anions contained in seawater samples without any disturbance due to matrices. Chemically bonded POE stationary phases retained not only anions but also cations. Anions were retained in the ion-exchange mode, although POE chains possess no ion exchange sites. The retention behavior suggested that eluent cations could be trapped among multiple POE chains via ion-dipole interaction, and that the trapped cations worked as the anion-exchange sites. Anions could be separated using crown ether, i.e., cyclic POE, as the eluent additive with a hydrophobic stationary phase, where analyte anions were retained via electrostatic interaction with the eluent cation trapped on the crown ether.


Expanding the disaster risk management framework: Measuring the constructed level of national identity as a factor of political risk.
Political risk is identified as a dominant risk category of disaster risk management (DRM) which could negatively affect the success of those measures implemented to reduce disaster risk. Key to political risk is the construct of national identity which, if poorly constructed, could greatly contribute to political risk. This article proposed a tool to measure the construct of national identity and to provide recommendations to strengthen the construct in order to mitigate the exacerbating influence it may have on political risk and ultimately on DRM. The design of the measurement tool consisted of a mixed methodological approach employing both quantitative and qualitative data. The data collection instruments included a literature review (which is shortly provided in the previous sections) and an empirical study that utilised data obtained through structured questionnaires. Although the results of the proposed measuring instrument did not include a representative sample of all the cultures in South Africa, the results alluded to different levels for the construction of national identity among black and white respondents, possibly because of different ideological expectations among these groups. The results of the study should be considered as a validation of the measuring tool and not necessarily of the construct of national identity in South Africa. The measuring tool is thus promising for future studies to reduce political risk and ultimately disaster risk.


The reliability of the Glasgow Coma Scale: a systematic review.
The Glasgow Coma Scale (GCS) provides a structured method for assessment of the level of consciousness. Its derived sum score is applied in research and adopted in intensive care unit scoring systems. Controversy exists on the reliability of the GCS. The aim of this systematic review was to summarize evidence on the reliability of the GCS. A literature search was undertaken in MEDLINE, EMBASE and CINAHL. Observational studies that assessed the reliability of the GCS, expressed by a statistical measure, were included. Methodological quality was evaluated with the consensus-based standards for the selection of health measurement instruments checklist and its influence on results considered. Reliability estimates were synthesized narratively. We identified 52 relevant studies that showed significant heterogeneity in the type of reliability estimates used, patients studied, setting and characteristics of observers. Methodological quality was good (n = 7), fair (n = 18) or poor (n = 27). In good quality studies, kappa values were ≥0.6 in 85%, and all intraclass correlation coefficients indicated excellent reliability. Poor quality studies showed lower reliability estimates. Reliability for the GCS components was higher than for the sum score. Factors that may influence reliability include education and training, the level of consciousness and type of stimuli used. Only 13% of studies were of good quality and inconsistency in reported reliability estimates was found. Although the reliability was adequate in good quality studies, further improvement is desirable. From a methodological perspective, the quality of reliability studies needs to be improved. From a clinical perspective, a renewed focus on training/education and standardization of assessment is required.


Embodied changes and the search for gynecological cancer diagnosis.
The detection and successful treatment of cancers is dependent on timely presentation with abnormal and often subtle symptoms. In this article, we draw on research conducted with Australian immigrant women in 2001-02 who experienced delays in diagnosis of gynecological cancer. Data from in-depth interviews with women with gynecological cancer indicated a common trajectory of an "illness career" whereby the search for diagnosis was often painful and lengthy, either because women normalized the abnormal signs or because their experiences of bodily abnormality, pain, and dysfunction contrasted with medical explanations. This delay was sometimes exacerbated by structural barriers. As a result, diagnosis was often protracted, during which time women presented with symptoms that were dismissed or resulted in the misdiagnosis, with a resultant loss of confidence in clinical services.


Prenatal detection of trisomy 21 by second-trimester ultrasound examination and maternal age in a non-selected population of 49 314 births in Norway.
To assess the contribution of the second-trimester routine ultrasound examination and maternal age (>or= 38 years) to the prenatal detection of trisomy 21 in a large non-selected population in which no other screening methods were carried out. A prospective follow-up study was carried out in a non-selected Norwegian population from 1987 to 2004, including a total of 49 314 births. Data from all cases of trisomy 21, detected prenatally by karyotyping following amniocentesis performed for abnormal ultrasound findings or because of maternal age >or= 38 years, or postnatally after delivery, were registered. The study period was divided into three 6-year periods. Eighty-eight cases of trisomy 21 were registered. The prenatal detection rate was 43% (38/88). No significant change in the detection rate was observed over the 18 years. Fourteen percent (12/88) of cases were detected as a result of advanced maternal age and 30% (26/88) by prenatal ultrasound examination. Of all women with a trisomy 21 fetus, 72% (63/88) were under 38 years of age. The percentage of women >or= 38 years opting for karyotyping during the three time periods decreased significantly from 51% to 50% and 36%, respectively. The termination rate of trisomy 21 fetuses was 84%, with no significant change over time. Our data can be considered as a reference standard for population screening for trisomy 21 based solely on maternal age and second-trimester ultrasound imaging. The prenatal detection rate of trisomy 21 cases was poor and remained unchanged throughout the 18-year study period. If improvement in detection rates is desired, additional programs are necessary.


Hepatic SRC-1 activity orchestrates transcriptional circuitries of amino acid pathways with potential relevance for human metabolic pathogenesis.
Disturbances in amino acid metabolism are increasingly recognized as being associated with, and serving as prognostic markers for chronic human diseases, such as cancer or type 2 diabetes. In the current study, a quantitative metabolomics profiling strategy revealed global impairment in amino acid metabolism in mice deleted for the transcriptional coactivator steroid receptor coactivator (SRC)-1. Aberrations were hepatic in origin, because selective reexpression of SRC-1 in the liver of SRC-1 null mice largely restored amino acids concentrations to normal levels. Cistromic analysis of SRC-1 binding sites in hepatic tissues confirmed a prominent influence of this coregulator on transcriptional programs regulating amino acid metabolism. More specifically, SRC-1 markedly impacted tyrosine levels and was found to regulate the transcriptional activity of the tyrosine aminotransferase (TAT) gene, which encodes the rate-limiting enzyme of tyrosine catabolism. Consequently, SRC-1 null mice displayed low TAT expression and presented with hypertyrosinemia and corneal alterations, 2 clinical features observed in the human syndrome of TAT deficiency. A heterozygous missense variant of SRC-1 (p.P1272S) that is known to alter its coactivation potential, was found in patients harboring idiopathic tyrosinemia-like disorders and may therefore represent one risk factor for their clinical symptoms. Hence, we reinforce the concept that SRC-1 is a central factor in the fine orchestration of multiple pathways of intermediary metabolism, suggesting it as a potential therapeutic target that may be exploitable in human metabolic diseases and cancer.


Long-term costs and health impact of continued global fund support for antiretroviral therapy.
By the end of 2011 Global Fund investments will be supporting 3.5 million people on antiretroviral therapy (ART) in 104 low- and middle-income countries. We estimated the cost and health impact of continuing treatment for these patients through 2020. Survival on first-line and second-line ART regimens is estimated based on annual retention rates reported by national AIDS programs. Costs per patient-year were calculated from country-reported ARV procurement prices, and expenditures on laboratory tests, health care utilization and end-of-life care from in-depth costing studies. Of the 3.5 million ART patients in 2011, 2.3 million will still need treatment in 2020. The annual cost of maintaining ART falls from $1.9 billion in 2011 to $1.7 billion in 2020, as a result of a declining number of surviving patients partially offset by increasing costs as more patients migrate to second-line therapy. The Global Fund is expected to continue being a major contributor to meeting this financial need, alongside other international funders and domestic resources. Costs would be $150 million less in 2020 with an annual 5% decline in first-line ARV prices and $150-370 million less with a 5%-12% annual decline in second-line prices, but $200 million higher in 2020 with phase out of stavudine (d4T), or $200 million higher with increased migration to second-line regimens expected if all countries routinely adopted viral load monitoring. Deaths postponed by ART correspond to 830,000 life-years saved in 2011, increasing to around 2.3 million life-years every year between 2015 and 2020. Annual patient-level direct costs of supporting a patient cohort remain fairly stable over 2011-2020, if current antiretroviral prices and delivery costs are maintained. Second-line antiretroviral prices are a major cost driver, underscoring the importance of investing in treatment quality to improve retention on first-line regimens.


Topology of surfaces for molecular Stark energy, alignment, and orientation generated by combined permanent and induced electric dipole interactions.
We show that combined permanent and induced electric dipole interactions of linear polar and polarizable molecules with collinear electric fields lead to a sui generis topology of the corresponding Stark energy surfaces and of other observables - such as alignment and orientation cosines - in the plane spanned by the permanent and induced dipole interaction parameters. We find that the loci of the intersections of the surfaces can be traced analytically and that the eigenstates as well as the number of their intersections can be characterized by a single integer index. The value of the index, distinctive for a particular ratio of the interaction parameters, brings out a close kinship with the eigenproperties obtained previously for a class of Stark states via the apparatus of supersymmetric quantum mechanics.


Acoustically tuned erbium-doped fiber ring laser.
We report what is to our knowledge the first narrow-linewidth, continuously tunable laser that uses an acousto-optic filter to achieve a broad tuning range. The unidirectional ring-laser configuration incorporates an erbium-doped fiber amplifier as the gain medium and an integrated, frequency-shift-compensated acousto-optic filter as the tuning element. Using a 980-nm optical pump with 60 mW of launched power, we obtained nearly 2 mW of laser output with 10% output coupling. A tuning range of 40 nm was achieved, centered about 1545 nm. Single-longitudinal-mode operation with a 10-kHz linewidth was demonstrated, although the laser wavelength hopped between nearby longitudinal modes because of path-length fluctuations. Wavelength switching at rates up to 100 kHz was examined.


Pseudomonas stutzeri YPL-1 Genetic Transformation and Antifungal Mechanism against Fusarium solani, an Agent of Plant Root Rot.
An actively antagonistic bacterium that could be used as a biocontrol agent against Fusarium solani, which causes root rots with considerable losses in many important crops, was isolated from a ginseng rhizosphere and identified as a strain of Pseudomonas stutzeri. In several biochemical tests with culture filtrates of P. stutzeri YPL-1 and in mutational analyses of antifungal activities of reinforced or defective mutants, we found that the anti-F. solani mechanism of the bacterium may involve a lytic enzyme rather than a toxic substance or antibiotic. P. stutzeri YPL-1 produced extracellular chitinase and laminarinase when grown on different polymers such as chitin, laminarin, or F. solani mycelium. These lytic extracellular enzymes markedly inhibited mycelial growth rather than spore germination and also caused lysis of F. solani mycelia and germ tubes. Scanning electron microscopy revealed degradation of the F. solani mycelium. Abnormal hyphal swelling and retreating were caused by the lysing agents from P. stutzeri YPL-1, and a penetration hole was formed on the hyphae in the region of interaction with the bacterium; the walls of this region were rapidly lysed, causing leakage of protoplasm. Genetically bred P. stutzeri YPL-1 was obtained by transformation of the bacterium with a broad-host-range vector, pKT230. Also, the best conditions for the transformation were investigated.


Immunogenetic studies of maternal-fetal relationships: a review: why newborn rhesus monkeys don't get hemolytic disease.
The discovery of the Rh blood group factor in humans was made using the red blood cells of rhesus monkeys. Because of its importance to human medicine and immunogenetics, this finding contributed greatly to the appreciation of the importance of nonhuman primates in research. It is now widely recognized that blood group incompatibility between mother and fetus can lead to differential fertility, fetal death, and hemolytic disease of the newborn (HDN). The blood group systems of several nonhuman primate species have been studied in detail and found to be analogous, although not identical, to those of humans. It is therefore surprising that HDN has been reported in only four nonhuman primate species--marmosets, sacred baboons, chimpanzees, and orangutans. Maternal-fetal blood group incompatibility and its consequences have been extensively studied in rhesus monkeys, and these macaques may well be representative of many nonhuman primates. Rhesus monkeys exhibit all five of the conditions that lead to HDN in humans: (1) blood group incompatible matings; (2) transplacental hemorrhage; (3) maternal immunization to blood group alloantigens on fetal erythrocytes; (4) transplacental transfer of maternal antibodies; and (5) coating of the newborn's erythrocytes. Yet, newborns show no clinical or hematological evidence of HDN. We have shown that the rhesus alloantibodies engendered by transplacental immunization do not mediate immune elimination of the newborn's erythrocytes. Evaluation of the maternal antibodies demonstrated that they have low titers and low avidities and perhaps belong to IgG subclasses that do not bind effectively to receptors on phagocytic cells of the rhesus reticuloendothelial system. The newborn's genotype may also affect the expression of allogeneic blood group antigens and thereby help protect the newborn's cells from destruction. These factors together undoubtedly play a major role in the survival of the antibody-coated newborn's RBC and are thus able to account for the absence of HDN in this species.


E2F-4 switches from p130 to p107 and pRB in response to cell cycle reentry.
The E2F transcription factor couples the coordinate expression of cell cycle proteins to their appropriate transition points. Its activity is controlled by the cell cycle regulators pRB, p107, and p130. These bind to E2F at defined but distinct stages of the cell cycle. Using specific antisera, we have identified the DP and E2F components of each of these species. Although present at very different levels, DP-1 and DP-2 are evenly distributed among each of these complexes. In contrast, the individual E2Fs have distinctly different binding profiles. Consistent with previous studies, E2F-1, E2F-2, and E2F-3 bind specifically to the retinoblastoma protein. In each case, their expression and DNA binding activity are restricted to post-G1/S fractions. Surprisingly, E2F-1 and E2F-3 make unequal contributions to the pRB-associated and free E2F activity, suggesting that these proteins perform different cell cycle functions. Most significantly, this study showed E2F-4 accounts for the vast majority of the endogenous E2F activity. In arrested cells, E2F-4 is sequestered by the p130 protein. However, as the cells pass the G1-to-S transition, the levels of pRB and p107 increase and E2F-4 now associates with both of these regulators. Despite this, a considerable amount of E2F-4 exists as free E2F. In G1 cells, this accounts for almost all of the free activity. Once the cells enter S phase, free E2F is composed of an equal mixture of E2F-4 and E2F-1.


Anti-obesity effects of alpha-lipoic acid mediated by suppression of hypothalamic AMP-activated protein kinase.
AMP-activated protein kinase (AMPK) functions as a fuel sensor in the cell and is activated when cellular energy is depleted. Here we report that alpha-lipoic acid (alpha-LA), a cofactor of mitochondrial enzymes, decreases hypothalamic AMPK activity and causes profound weight loss in rodents by reducing food intake and enhancing energy expenditure. Activation of hypothalamic AMPK reverses the effects of alpha-LA on food intake and energy expenditure. Intracerebroventricular (i.c.v.) administration of glucose decreases hypothalamic AMPK activity, whereas inhibition of intracellular glucose utilization through the administration of 2-deoxyglucose increases hypothalamic AMPK activity and food intake. The 2-deoxyglucose-induced hyperphagia is reversed by inhibiting hypothalamic AMPK. Our findings indicate that hypothalamic AMPK is important in the central regulation of food intake and energy expenditure and that alpha-LA exerts anti-obesity effects by suppressing hypothalamic AMPK activity.


Transapical transcatheter aortic valve implantation vs conventional aortic valve replacement in high-risk patients with previous cardiac surgery: a propensity-score analysis.
The present analysis compared clinical and mid-term outcomes of patients with previous cardiac surgery undergoing transapical transcatheter aortic valve implantation (TAVI) with propensity-matched patients undergoing conventional redo aortic valve replacement (cAVR). Since 2008, 508 patients were treated with TAVI. Fifty-three of these patients presented with a history of cardiac surgery and underwent transapical TAVI using the Edwards SAPIEN bioprosthesis. A propensity-matched control group of 53 patients receiving cAVR was generated out of the hospital's database. The mean age for all the patients was 77.8 ± 4.5 years. The logistic EuroSCORE was 28.4 ± 13.6% in mean, and mean EuroSCORE II was 8.56 ± 3.93%. The mean follow-up time was 245 ± 323 days, which equated to a total of 700 patient-months. The observed hospital mortality did not differ significantly between TAVI and cAVR (TAVI: 9.4% and cAVR: 5.7%; P = 0.695). Six-month survival was 83.0% for the TAVI and 86.8% for the cAVR patients (P = 0.768). Postoperative bleedings (TAVI: 725 ± 1770 ml and cAVR: 1884 ± 6387; P = 0.022), the need for transfusion (TAVI: 1.7 ± 5.3 vs cAVR: 6.2 ± 13.7 units packed red blood cells (PRBC); P = 0.030), consecutive rethoracotomy (TAVI: 1.9% vs cAVR: 16.9%; P = 0.002) and postoperative delirium (TAVI: 11.5% vs cAVR: 28.3%; P = 0.046) were more common in the cAVR patients. The TAVI patients suffered more frequently from respiratory failure (TAVI: 11.3% vs cAVR: 0.0%; P = 0.017) and mean grade of paravalvular regurgitation (TAVI: 0.8 ± 0.2 vs cAVR: 0.0; P = 0.047). Although primary ventilation time (P = 0.020) and intensive care unit stay (P = 0.022) were shorter in the TAVI patients, mean hospital stay did not differ significantly (P = 0.108). Transapical TAVI as well as surgical aortic valve replacement provided good clinical results. The pattern of postoperative morbidity and mortality was different for both entities, but the final clinical outcome did not differ significantly. Both techniques can be seen as complementary approaches by means of developing a tailor-made and patient-orientated surgery.


Somite Division and New Boundary Formation by Mechanical Strain.
Somitogenesis, the primary segmentation of the vertebrate embryo, is associated with oscillating genes that interact with a wave of cell differentiation. The necessity of cell-matrix adherence and embryonic tension, however, suggests that mechanical cues are also involved. To explicitly investigate this, we applied surplus axial strain to live chick embryos. Despite substantial deformations, the embryos developed normally and somite formation rate was unaffected. Surprisingly, however, we observed slow cellular reorganizations of the most elongated somites into two or more well-shaped daughter somites. In what appeared to be a regular process of boundary formation, somites divided and fibronectin was deposited in between. Cell counts and morphology indicated that cells from the somitocoel underwent mesenchymal-epithelial transition; this was supported by a Cellular Potts model of somite division. Thus, although somitogenesis appeared to be extremely robust, we observed new boundary formation in existing somites and conclude that mechanical strain can be morphologically instructive.


Radiation hybrid map, physical map, and low-pass genomic sequence of the canine prcd region on CFA9 and comparative mapping with the syntenic region on human chromosome 17.
Progressive rod-cone degeneration (prcd) is a canine retinal disease that maps to the centromeric end of CFA9 in a region of synteny with the distal part of HSA17q. As such, prcd has been postulated as the only animal model of RP17, a human retinitis pigmentosa locus that maps to 17q22. In an effort to establish more detailed regions of synteny between dog CFA9 and the HSA17q-ter region, we created a robust gene-enriched CFA9-RH08(3000) map with 34 gene-based markers and 12 microsatellites, with the highest resolution and number of markers for the centromeric end of CFA9. Furthermore, we built an approximately 1.5-Mb physical map containing both GRB2 and GALK1, genes so far identified by meiotic linkage analysis as being closest to the prcd locus, and generated about 1.2 Mb low-pass (3.2x) canine sequence. Canine to human comparative sequence analysis identified 49 transcripts that had been previously mapped to the HSA17q25 region. The generated low-pass canine sequence was annotated with a working draft of human sequence from HSA17q25, and we used this scaffold to order and orient the canine sequence against human. This order and orientation are preliminary, as high-throughput genomic sequencing of HSA17q-ter has not been fully completed.


Incidence and Trends of Sepsis in US Hospitals Using Clinical vs Claims Data, 2009-2014.
Estimates from claims-based analyses suggest that the incidence of sepsis is increasing and mortality rates from sepsis are decreasing. However, estimates from claims data may lack clinical fidelity and can be affected by changing diagnosis and coding practices over time. To estimate the US national incidence of sepsis and trends using detailed clinical data from the electronic health record (EHR) systems of diverse hospitals. Retrospective cohort study of adult patients admitted to 409 academic, community, and federal hospitals from 2009-2014. Sepsis was identified using clinical indicators of presumed infection and concurrent acute organ dysfunction, adapting Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3) criteria for objective and consistent EHR-based surveillance. Sepsis incidence, outcomes, and trends from 2009-2014 were calculated using regression models and compared with claims-based estimates using International Classification of Diseases, Ninth Revision, Clinical Modification codes for severe sepsis or septic shock. Case-finding criteria were validated against Sepsis-3 criteria using medical record reviews. A total of 173 690 sepsis cases (mean age, 66.5 [SD, 15.5] y; 77 660 [42.4%] women) were identified using clinical criteria among 2 901 019 adults admitted to study hospitals in 2014 (6.0% incidence). Of these, 26 061 (15.0%) died in the hospital and 10 731 (6.2%) were discharged to hospice. From 2009-2014, sepsis incidence using clinical criteria was stable (+0.6% relative change/y [95% CI, -2.3% to 3.5%], P = .67) whereas incidence per claims increased (+10.3%/y [95% CI, 7.2% to 13.3%], P < .001). In-hospital mortality using clinical criteria declined (-3.3%/y [95% CI, -5.6% to -1.0%], P = .004), but there was no significant change in the combined outcome of death or discharge to hospice (-1.3%/y [95% CI, -3.2% to 0.6%], P = .19). In contrast, mortality using claims declined significantly (-7.0%/y [95% CI, -8.8% to -5.2%], P < .001), as did death or discharge to hospice (-4.5%/y [95% CI, -6.1% to -2.8%], P < .001). Clinical criteria were more sensitive in identifying sepsis than claims (69.7% [95% CI, 52.9% to 92.0%] vs 32.3% [95% CI, 24.4% to 43.0%], P < .001), with comparable positive predictive value (70.4% [95% CI, 64.0% to 76.8%] vs 75.2% [95% CI, 69.8% to 80.6%], P = .23). In clinical data from 409 hospitals, sepsis was present in 6% of adult hospitalizations, and in contrast to claims-based analyses, neither the incidence of sepsis nor the combined outcome of death or discharge to hospice changed significantly between 2009-2014. The findings also suggest that EHR-based clinical data provide more objective estimates than claims-based data for sepsis surveillance.


A new video pachometer.
Many studies of the cornea would benefit from a simple, objective method to measure corneal thickness. In this study, a new optical pachometer based on video and computer technology was designed and tested. The slit beam of a photographic slit lamp was monitored with a video camera through one half of the biomicroscope. When the slit was properly aligned with the cornea, the operator triggered a flash, and one video frame that included the flash was captured. A custom software package detected epithelial and endothelial edges. Corneal thickness was calculated from the median corneal image width and image widths from similar measurements of contact lenses with known thicknesses. Corneal thickness was measured in 25 subjects by using this new instrument and was compared to thickness measured by using a conventional Haag-Streit pachometer. Corneal thickness in the 25 subjects measured on the new instrument was 512+/-20 microm and 515-/+21 microm in the right and left eyes, respectively (mean+/-SD). Thickness of the same corneas measured on the Haag-Streit pachometer was 530+/-22 microm and 534+/-20 microm in right and left eyes, respectively. The average SD of 10 consecutive measurements was 6.6 microm and 6.7 microm on the video and Haag-Streit pachometers, respectively (n = 50 corneas). The video pachometer provides a fast, objective means of measuring corneal thickness. It is simple to use and provides precision equal to that of the Haag-Streit pachometer.


Significance of premature stop codons in env of simian immunodeficiency virus.
The location of the translational termination codon for the transmembrane protein (TMP) varies in three infectious molecular clones of simian immunodeficiency virus from macaques (SIVmac). The SIVmac251 and SIVmac142 infectious clones have premature stop signals that differ in location by one codon; transfection of these DNAs into human HUT-78 cells yielded virus with a truncated TMP (28 to 30 kilodaltons [kDa]). The SIVmac239 infectious clone does not have a premature stop codon in its TMP-coding region. Transfection of HUT-78 cells with this clone initially yielded virus with a full-length TMP (41 kDa). At 20 to 30 days posttransfection, SIVmac239 virus with a 41-kDa TMP gradually disappeared coincident with the emergence of a virus with a 28-kDa TMP. Virus production dramatically increased in parallel with the emergence of a virus with a 28-kDa TMP. Sequence analysis of viral DNAs from these cultures showed that premature stop codons arising by point mutation were responsible for the change in size of the TMP with time. A similar selective pressure for truncated forms of TMP was observed when the SIVmac239 clone was transfected into human peripheral blood lymphocytes (PBL). In contrast, no such selective pressure was observed in macaque PBL. When the SIVmac239 clone was transfected into macaque PBL and the resultant virus was serially passaged in macaque PBL, the virus replicated very well and maintained a 41-kDa TMP for 80 days in culture. Macaque monkeys were infected with SIVmac239 having a 28-kDa TMP; virus subsequently recovered from T4-enriched lymphocytes of peripheral blood showed only the 41-kDa form of TMP. These results indicate that the natural form of TMP in SIVmac is the full-length 41-kDa TMP, just as in human immunodeficiency virus type 1. Viruses with truncated forms of TMP appear to result from mutation and selection during propagation in unnatural human cells.


Competitive dominance in plant communities: Modeling approaches and theoretical predictions.
Quantitative predictions about the processes that promote species coexistence are a subject of active research in ecology. In particular, competitive interactions are known to shape and maintain ecological communities, and situations where some species out-compete or dominate over some others are key to describe natural ecosystems. Here we develop ecological theory using a stochastic, synthetic framework for plant community assembly that leads to predictions amenable to empirical testing. We propose two stochastic, continuous-time Markov models that incorporate competitive dominance through a hierarchy of species heights. The first model, which is spatially implicit, predicts both the expected number of species that survive and the conditions under which heights are clustered in realized model communities. The second one allows spatially-explicit interactions of individuals and alternative mechanisms that can help shorter plants overcome height-driven competition, and it demonstrates that clustering patterns remain, not only locally but also across increasing spatial scales. Moreover, although plants are actually height-clustered in the spatially-explicit model, plant species abundances are not necessarily skewed to taller plants.


Conserved mechanism of Wnt signaling function in the specification of vulval precursor fates in C. elegans and C. briggsae.
The C. elegans hermaphrodite vulva serves as a paradigm for understanding how signaling pathways control organ formation. Previous studies have shown that Wnt signaling plays important roles in vulval development. To understand the function and evolution of Wnt signaling in Caenorhabditis nematodes we focused on C. briggsae, a species that is substantially divergent from C. elegans in terms of the evolutionary time scale yet shares almost identical morphology. We isolated mutants in C. briggsae that display multiple pseudo-vulvae resulting from ectopic VPC induction. We cloned one of these loci and found that it encodes an Axin homolog, Cbr-PRY-1. Our genetic studies revealed that Cbr-pry-1 functions upstream of the canonical Wnt pathway components Cbr-bar-1 (beta-catenin) and Cbr-pop-1(tcf/lef) as well as the Hox target Cbr-lin-39 (Dfd/Scr). We further characterized the pry-1 vulval phenotype in C. briggsae and C. elegans using 8 cell fate markers, cell ablation, and genetic interaction approaches. Our results show that ectopically induced VPCs in pry-1 mutants adopt 2° fates independently of the gonad-derived inductive and LIN-12/Notch-mediated lateral signaling pathways. We also found that Cbr-pry-1 mutants frequently show a failure of P7.p induction. A similar, albeit low penetrant, defect is also observed in C. elegans pry-1 mutants. The genetic analysis of the P7.p induction defect revealed that it was caused by altered regulation of lin-12 and its transcriptional target lip-1 (MAP kinase phosphatase). Thus, our results provide evidence for LIN-12/Notch-dependent and independent roles of Wnt signaling in promoting 2 degrees VPC fates in both nematode species.


A newly classified vertebrate calpain protease, directly ancestral to CAPN1 and 2, episodically evolved a restricted physiological function in placental mammals.
The most studied members of the calpain protease superfamily are CAPN1 and 2, which are conserved across vertebrates. Another similar family member called mu/m-CAPN has been identified in birds alone. Here, we establish that mu/m-CAPN shares one-to-one orthology with CAPN11, previously described only in eutherians (placental mammals). We use the name CAPN11 for this family member and identify orthologues across vertebrate lineages, which form a monophyletic phylogenetic clade directly ancestral to CAPN1 and 2. In lineages branching before therians (live-bearing mammals), the CAPN11 coding region has evolved under strong purifying selection, with low nonsynonymous (d(N)) versus synonymous (d(S)) substitution rates (d(N)/d(S) = 0.076 across pretherians), and its transcripts were detected widely across different tissues. These characteristics are present in CAPN1 and 2 across vertebrate lineages and indicate that pretherian CAPN11 likewise has conserved a wide physiological function. However, an approximately 7-fold elevation in d(N)/d(S) is evident along the CAPN11 branch splitting eutherians from platypus, paralleled by a shift to "testis-specific" gene regulation. Estimates of d(N)/d(S) in eutherians were approximately 3-fold elevated compared with pretherians and coding and transcriptional-level evidence suggests that CAPN11 is functionally absent in marsupials. Many CAPN11 sites are functionally constrained in eutherians to conserve a residue with radically different biochemical properties to a fixed state shared between pretherian CAPN11 and CAPN1 and 2. Protein homology modeling demonstrated that many such eutherian-specific residue replacements modify or ablate interactions with the calpain inhibitor calpastatin that are observed in both pretherian orthologues and CAPN1/2. We propose a model akin to the Dykhuizen-Hartl effect, where inefficient purifying selection and increased genetic drift associated with a reduction in effective population size, drove the fixation of mutations in regulatory and coding regions of CAPN11 of a common marsupial-eutherian ancestor. A subset of these changes had a cumulative adaptive advantage in a eutherian ancestor because of lineage-specific aspects of sperm physiology, whereas in marsupials, no advantage was realized and the gene was disabled. This work supports that functional divergence among gene family member orthologues is possible in the absence of widespread positive selection.


Detecting mitoses in time-lapse images of embryonic epithelia using intensity analysis.
Although the frequency and orientation of mitoses can significantly affect the mechanics of early embryo development, these data have not been available due to a shortage of suitable automated techniques. Fluorescence imaging, though popular, requires biochemical intervention and is not always possible or desirable. Here, a new technique that takes advantage of a localized intensity change that occurs in bright field images is used to identify mitoses. The algorithm involves mapping a deformable, sub-cellular triangular mesh from one time-lapse image to the next so that corresponding regions can be identified. Triangles in the mesh that undergo darkening of a sufficient degree over a period consistent with mitosis are flagged. Mitoses are assumed to occur along the short axis of elliptical areas fit to suitably sized clusters of flagged triangles. The algorithm is less complex than previous approaches and it has strong discrimination characteristics. When applied to 15 image sets from neurulation-stage axolotl (Ambystoma mexicanum) embryos, it was able to correctly detect 86% of the manually identified mitoses, had less than 5% false positives and produced average angular errors of only 15 degrees . The new algorithm is simpler to implement than those previously available, is substantially more accurate, and provides data that is important for understanding the mechanics of morphogenetic movements.


Type 2 immunity in asthma.
Type 2-immunity represents the typical adaptive response to allergen exposure in atopic individuals. It mainly involves Th2 cells and immunoglobulin E, as the main orchestrators of type 2-inflammation. Recently, it has been highlighted that allergens may be responsible for a Th2 response beside specific IgE activation and that a number of other environmental stimuli, such as viruses and pollutants, can trigger the same pattern of inflammation beyond atopy. Emerging data sustain a substantial role of the so-called epithelial dysfunction in asthma pathogenesis, both from anatomic and functional point of view. Furthermore an increasing amount of evidence demonstrates the relevance of innate immunity in polarizing a Th2 impaired response in asthmatic patients. Under this perspective, the complex cross-talking between airway epithelium, innate and adaptive immunity is emerging as a major determinant of type 2-inflammation beyond allergens. This review will include an update on the relevance of dysregulation of innate and adaptive type 2-immunity in asthma pathogenesis, particularly severe asthma, and on the role of the allergens that are associated with severe asthma. Type 2-immunity also will be reviewed in the light of the current and upcoming targeted treatments for severe asthma.


Quantification of peptide m/z distributions from 13C-labeled cultures with high-resolution mass spectrometry.
Isotopic labeling studies of primary metabolism frequently utilize GC/MS to quantify (13)C in protein-hydrolyzed amino acids. During processing some amino acids are degraded, which reduces the size of the measurement set. The advent of high-resolution mass spectrometers provides a tool to assess molecular masses of peptides with great precision and accuracy and computationally infer information about labeling in amino acids. Amino acids that are isotopically labeled during metabolism result in labeled peptides that contain spatial and temporal information that is associated with the biosynthetic origin of the protein. The quantification of isotopic labeling in peptides can therefore provide an assessment of amino acid metabolism that is specific to subcellular, cellular, or temporal conditions. A high-resolution orbital trap was used to quantify isotope labeling in peptides that were obtained from unlabeled and isotopically labeled soybean embryos and Escherichia coli cultures. Standard deviations were determined by estimating the multinomial variance associated with each element of the m/z distribution. Using the estimated variance, quantification of the m/z distribution across multiple scans was achieved by a nonlinear fitting approach. Observed m/z distributions of uniformly labeled E. coli peptides indicated no significant differences between observed and simulated m/z distributions. Alternatively, amino acid m/z distributions obtained from GC/MS were convolved to simulate peptide m/z distributions but resulted in distinct profiles due to the production of protein prior to isotopic labeling. The results indicate that peptide mass isotopologue measurements faithfully represent mass distributions, are suitable for quantification of isotope-labeling-based studies, and provide additional information over existing methods.


Multimodal Breast Phantoms for Microwave, Ultrasound, Mammography, Magnetic Resonance and Computed Tomography Imaging.
The aim of this work was to develop multimodal anthropomorphic breast phantoms suitable for evaluating the imaging performance of a recently-introduced Microwave Imaging (MWI) technique in comparison to the established diagnostic imaging modalities of Magnetic Resonance Imaging (MRI), Ultrasound (US), mammography and Computed Tomography (CT). MWI is an emerging technique with significant potential to supplement established imaging techniques to improve diagnostic confidence for breast cancer detection. To date, numerical simulations have been used to assess the different MWI scanning and image reconstruction algorithms in current use, while only a few clinical trials have been conducted. To bridge the gap between the numerical simulation environment and a more realistic diagnostic scenario, anthropomorphic phantoms which mimic breast tissues in terms of their heterogeneity, anatomy, morphology, and mechanical and dielectric characteristics, may be used. Key in this regard is achieving realism in the imaging appearance of the different healthy and pathologic tissue types for each of the modalities, taking into consideration the differing imaging and contrast mechanisms for each modality. Suitable phantoms can thus be used by radiologists to correlate image findings between the emerging MWI technique and the more familiar images generated by the conventional modalities. Two phantoms were developed in this study, representing difficult-to-image and easy-to-image patients: the former contained a complex boundary between the mammary fat and fibroglandular tissues, extracted from real patient MRI datasets, while the latter contained a simpler and less morphologically accurate interface. Both phantoms were otherwise identical, with tissue-mimicking materials (TMMs) developed to mimic skin, subcutaneous fat, fibroglandular tissue, tumor and pectoral muscle. The phantoms' construction used non-toxic materials, and they were inexpensive and relatively easy to manufacture. Both phantoms were scanned using conventional modalities (MRI, US, mammography and CT) and a recently introduced MWI radar detection procedure called in-coherent Multiple Signal Classification (I-MUSIC). Clinically realistic artifact-free images of the anthropomorphic breast phantoms were obtained using the conventional imaging techniques as well as the emerging technique of MWI.


North Carolina physician-based preventive oral health services improve access and use among young Medicaid enrollees.
To combat disparities in oral health and access to dental care among infants and toddlers, most state Medicaid programs now reimburse physician-based preventive oral health services such as fluoride varnish applications. We used geospatial data to examine the distribution of dental and medical Medicaid providers of pediatric oral health services throughout North Carolina to determine if these services have improved access to care for Medicaid enrollees younger than age three. We then used claims data to examine the association between distance from these practices and use of dental services for a cohort of approximately 1,000 young children. Among one hundred counties, four counties had no physician-based preventive oral health services, and nine counties had no dental practice. While children who lived farther from the nearest dental practice were less likely to make dental visits, distance from physician-based preventive oral health services did not predict utilization. For young Medicaid enrollees, oral health services provided in medical offices can improve access and increase utilization.


[Significance of psychiatrists' personal variables in treatment of persons suffering from schizophrenia].
The aim of the study is to investigate the significance of psychiatrists' personal variables in treatment and functioning of persons suffering from schizophrenia. 150 patients with schizophrenia indicated by 30 psychiatrists were included in the study. Data was collected during single interview with the use of Dyadic Questionnaire of the Therapist-Patient Relationship, Questionnaire of Pharmacological Treatment Assessment, Life Satisfaction Scale, Emotion Control Scale, Life Orientation Test, Overall Self-Efficacy Scale, Social Functioning Scale and Sociodemographic Questionnaire. Stepwise regression was used to determine the psychiatrist's related variables which explain variables associated with a patient. Psychiatrists' personal variables explained up to 20% of variance in social functioning of the patients with schizophrenia. Variables describing psychiatrists were most significant in explaining assessment of psychiatrist's professionalism and rejection of a psychiatrist by a patient. The abilities to identify medication symptoms and adherence to pharmacotherapy were only slightly explained by patient - psychiatrist collaboration length. Results indicate that psychiatrists' personal characteristics are significant for the development of the relationship between persons with schizophrenia and their psychiatrists.