Endorectal MR imaging and MR spectroscopic imaging for locally recurrent prostate cancer after external beam radiation therapy: preliminary experience.
To evaluate endorectal magnetic resonance (MR) imaging and MR spectroscopic imaging for the depiction of locally recurrent prostate cancer after external beam radiation therapy. Endorectal MR imaging and MR spectroscopic imaging were performed in 21 patients with biochemical failure after external beam radiation therapy for prostate cancer. Two readers independently and retrospectively reviewed MR images and rated the likelihood of recurrent tumor on a five-point scale. Spectroscopic voxels were considered suspicious for malignancy if the choline level was elevated and citrate was absent. Receiver operating characteristic curve analysis was used to assess cancer detection in each side of the prostate with endorectal MR imaging and spectroscopic imaging at different thresholds based on the scores assigned by the two readers and on the number of suspicious voxels in each hemiprostate, respectively. The presence or absence of cancer at subsequent transrectal biopsy was used as the standard of reference. Biopsy demonstrated locally recurrent prostate cancer in nine hemiprostates in six patients. The area under the receiver operating characteristic curve for the detection of locally recurrent cancer with MR imaging was 0.49 and 0.51 for readers 1 and 2, respectively. By using the number of suspicious voxels to define different diagnostic thresholds, the area under the receiver operating characteristic curve for MR spectroscopic imaging was significantly (P < .005) higher, at 0.81. In particular, the presence of three or more suspicious voxels in a hemiprostate showed a sensitivity and specificity of 89% and 82%, respectively, for the diagnosis of local recurrence. Seven hemiprostates demonstrated complete metabolic atrophy at spectroscopic imaging and only postirradiation atrophy at biopsy. Preliminary data suggest that MR spectroscopic imaging, but not endorectal MR imaging, may be of value for the depiction of locally recurrent prostate cancer after radiation therapy.


Piezosurgery in the excision of middle-ear tumors: effects on mineralized and non-mineralized tissues.
Piezosurgery is a new instrument able to cut bone without necrosis and non-mineralized tissue damage. The aim of this study was to test the Piezosurgery as a new and alternative method for the excision of middle-ear tumors and to analyze its effects on soft tissues. The Piezosurgery device was used to excise eight glomus tympanicum tumors and two primary B-cell lymphomas of the middle ear. The piezoelectric device uses low-frequency ultrasonic waves (24.7-29.5 kHz); the applied power can be modulated between 2.8 and 16 W and is programmed in accordance to the density of the bone. The equipment consists of two hand-pieces, two inserts, and two peristaltic pumps. The microvibrations that are created in the piezoelectric hand-piece cause the inserts to vibrate linearly by between 60 and 210 microm. Piezosurgery provided excellent control in all the patients, without bleeding and harmful effects on the adjacent structures of the middle and inner ear. Piezosurgery is a new and revolutionary osteotomy technique utilizing the microvibrations of scalpels at ultrasonic frequency, so that soft tissue will not be damaged even upon accidental contact with the cutting tip. The vibration frequency of Piezosurgery is optimal for mineralized tissue and does not cut the adjacent soft tissue, minimizing the risk of harming the adjacent tissues. This renders the piezoelectric device ideal for application at the border between hard and soft tissues as in the excision of a middle-ear tumor.


Structural and Functional Studies of the Daunorubicin Priming Ketosynthase DpsC.
Daunorubicin is a type II polyketide, one of a large class of polyaromatic natural products with anticancer, antibiotic, and antiviral activity. Type II polyketides are formed by the assembly of malonyl-CoA building blocks, though in rare cases, biosynthesis is initiated by the incorporation of a nonmalonyl derived starter unit, which adds molecular diversity to the poly-β-ketone backbone. Priming mechanisms for the transfer of novel starter units onto polyketide synthases (PKS) are still poorly understood. Daunorubicin biosynthesis incorporates a unique propionyl starter unit thought to be selected for by a subclass ("DpsC type") of priming ketosynthases (KS III). To date, however, no structural information exists for this subclass of KS III enzymes. Although selectivity for self-acylation with propionyl-CoA has previously been implied, we demonstrate that DpsC shows no discrimination for self-acylation or acyl-transfer to the cognate acyl carrier protein, DpsG with short acyl-CoAs. We present five crystal structures of DpsC, including apo-DpsC, acetyl-DpsC, propionyl-DpsC, butyryl-DpsC, and a cocrystal of DpsC with a nonhydrolyzable phosphopantetheine (PPant) analogue. The DpsC crystal structures reveal the architecture of the active site, the molecular determinants for catalytic activity and homology to O-malonyl transferases, but also indicate distinct differences. These results provide a structural basis for rational engineering of starter unit selection in type II polyketide synthases.


An all-silicon passive optical diode.
A passive optical diode effect would be useful for on-chip optical information processing but has been difficult to achieve. Using a method based on optical nonlinearity, we demonstrate a forward-backward transmission ratio of up to 28 decibels within telecommunication wavelengths. Our device, which uses two silicon rings 5 micrometers in radius, is passive yet maintains optical nonreciprocity for a broad range of input power levels, and it performs equally well even if the backward input power is higher than the forward input. The silicon optical diode is ultracompact and is compatible with current complementary metal-oxide semiconductor processing.


Effects of sulindac on renal function and prostaglandin synthesis in patients with moderate chronic renal insufficiency.
The renal effects of therapeutic doses of sulindac were studied in nine patients with stable renal insufficiency, mean creatinine clearance 37.0 +/- 2.2 ml min-1 1.73 m-2 (range 24.7-54.6 ml min-1 1.73 m-2). Nine days' treatment with sulindac produced a small, but significant, reduction in the mean creatinine clearance (37.0 +/- 2.2 to 34.7 +/- 2.2 ml min-1 1.73 m-2; P less than 0.02) and 99mTc diethylenetriaminepenta-acetate (DTPA) clearance (35.5 +/- 3.4 to 31.4 +/- 3.6 ml min-1 1.73 m-2; P less than 0.02) without altering body weight, effective renal plasma flow [131I]hippuran clearance), plasma renin activity (PRA), 24 h urinary volume or electrolyte excretion. After discontinuation of sulindac, creatinine clearance returned to pretreatment values. In five female patients, pretreatment urinary excretion of the 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha), a stable breakdown product of prostacyclin (PGI2), was significantly reduced (P less than 0.02) when compared with four healthy controls, whereas prostaglandin E2 (PGE2) was unchanged. Administration of sulindac did not significantly alter the excretion rate of PGE2 or 6-ketoPGF1 alpha in this group of patients. In chronic renal disease with moderate renal impairment, reduced renal prostacyclin synthesis may be an important predisposing factor to the renal toxicity associated with the use of non-steroidal anti-inflammatory drugs (NSAID). Short term use of sulindac in therapeutic doses does not appear to influence the excretion of prostaglandins and produces only a minor reversible change in renal function; used cautiously it may have advantages over other NSAID in these patients.


P19 embryonal carcinoma cells exhibit high sensitivity to botulinum type C and D/C mosaic neurotoxins.
Botulinum neurotoxins (BoNTs) inhibit neurotransmitter release at peripheral nerve terminals. They are serologically classified from A to G, C/D and D/C mosaic neurotoxins forming further subtypes of serotypes C and D. Cultured primary neurons, as well as neuronal cell lines such as PC12 and Neuro-2a, are often utilized in cell-based experiments on the toxic action of botulinum toxins. However, there are very few reports of the use of neural cell lines for studying BoNTs/C and D. In addition, the differentiated P19 neuronal cell line, which possesses cholinergic properties, has yet to be tested for its susceptibility to BoNTs. Here, the responsiveness of differentiated P19 cells to BoNT/C and BoNT/DC is reported. Both BoNT/C and BoNT/DC were shown to effectively bind to, and be internalized by, neurons derived from P19 cells. Subsequently, the intracellular substrates for BoNT/C and BoNT/DC were cleaved by treatment of the cells with the toxins in a ganglioside-dependent manner. Moreover, P19 neurons exhibited high sensitivity to BoNT/C and BoNT/DC, to the same extent as cultured primary neurons. These findings suggest that differentiated P19 cells possess full sensitivity to BoNT/C and BoNT/DC, thus making them a novel susceptible cell line for research into BoNTs.


[The productive district of textile asbestos in the Lower Iseo Lake area].
In the Bergamo area of Basso Sebino (lower Lake Iseo), for decades there has been a large concentration of small firms (mostly family-run), specialised in the production of rubber gaskets. Within this production field, some companies used to manufacture textile asbestos exclusively or as part of their business. The asbestos goods were therefore marketed as they were or subsequently cut and transformed into gaskets. Among the 5 companies involved, "Manifattura Colombo & C" was the first one that started this production in the district, and the one that engaged the greatest number of employees (considering both the Sarnico and Predore factories). In the Predore factory, operating from the fifties to 1979, the asbestos thread production was carried out exclusively and to the complete technological cycle (i.e. from crashing the raw asbestos that was brought in from Balangero). The whole process was performed in an extremely dusty environment (according to the witness of former workers), expecially during the first production steps. In the Sarnico factory which operated from 1920 to 1993, they produced textile asbestos items (laces, ropes, etc.), as well as rubber and metal gaskets. The latter were coupled with asbestos by means of metal-plastics co-moulding operations, in order to obtain gaskets highly resistant to exhaust vapours, gases, oils, solvents and so on. The environmental data available (referred to the 1980-1992 period), supply evidence of severe exposure in the first years of activity, whilst a sharp reduction in the asbestos-fiber concentration rate was achieved along the years, thanks mainly to the completion and improvement of exhaust systems installed on winding and braiding machines. Finally we shortly describe the work of the four other factories and in more detail that of the "Manifattura Sebina srl" is mentioned herein because, although this is a "typical" textile mill and exclusively manufacturing cotton products, a considerable number of cases of mesothelioma has been detected among its workforce. This has been attributed to the presence of asbestos insulated piping, to maintenance and replacement interventions on looms brake pads, and above all to the nearly 50 asbestos blankets that were employed in the weekly fire-fighting exercises, and usually leaned against the walls of departments, with no protection whatsoever.


Fibromuscular dysplasia and the brain. I. Observations on angiographic, clinical and genetic characteristics.
The angiographic, clinical, and genetic characteristics of fibromuscular dysplasia (FMD) are reviewed in 37 patients (mean age 48 years) selected from a pool of 4000 angiograms of carotid or vertebral arteries. FMD was a neglected pathogenic factor in 28 patients with hemorrhagic or ischemic cerebral lesions. The aneurysms found in 19 patients had conventional appearance and were mainly located in the internal carotid or middle cerebral arteries and on the same side as the most affected cervical artery, which suggests that aneurysms and FMD are pathogenically related. A clinical syndrome is presented where headache, ECG-abnormalities, hypertension, mental distress, tinnitus, vertigo, arrhythmia, TIA, and syncope are frequent components. Hemicrania, sometimes combined with ipsilateral Horner's Syndrome, was found in patients with advanced lesions in the carotid artery of the same side. An associated occurrence of stroke in pedigrees, especially among young and middle aged females, indicates that FMD in the majority of cases in inherited as an autosomal dominant trait with reduced penetrance in males.


Treating alcohol withdrawal with oral baclofen: a randomized, double-blind, placebo-controlled trial.
Abrupt cessation of alcohol intake causes habituated drinkers to experience symptoms of alcohol withdrawal syndrome (AWS). To determine the effect of the gamma-aminobutyric acid (GABA)-B agonist baclofen on the course of acute symptomatic AWS. Prospective, randomized, double-blind, placebo-controlled clinical study. Two tertiary-care hospitals in Duluth, Minnesota. Inpatient adults admitted for any reason (including AWS) judged to be at high risk for AWS. Inpatients who developed symptoms of AWS received symptom-triggered benzodiazepine treatment using lorazepam by standard protocol, and were randomized to receive baclofen 10 mg or placebo, 3 times per day, orally. AWS severity was assessed using the Clinical Institute Withdrawal Assessment of Alcohol Scale, Revised (CIWA-Ar); lorazepam dose was monitored. Seventy-nine subjects were enrolled. The 44 subjects who developed symptoms of AWS were randomized to baclofen or placebo. Thirty-one subjects (18 baclofen, 13 placebo) completed 72 hours of assessments, either entirely as inpatients or with outpatient follow-up. The need for high doses of benzodiazepines (20 mg or more of lorazepam over 72 hours) to control AWS was less likely in the baclofen treatment group (1 of 18) than in the placebo-treated group (7 of 13) (P = 0.004). We found that the use of baclofen was associated with a significant reduction in the use of high doses of benzodiazepine (lorazepam) in the management of symptomatic AWS. The use of low-dose baclofen in the management of AWS deserves further study, as reduced dependence on high-dose benzodiazepines in AWS management could improve patient safety.


Hematologic and serum biochemical reference intervals for free-ranging common bottlenose dolphins (Tursiops truncatus) and variation in the distributions of clinicopathologic values related to geographic sampling site.
To develop robust reference intervals for hematologic and serum biochemical variables by use of data derived from free-ranging bottlenose dolphins (Tursiops truncatus) and examine potential variation in distributions of clinicopathologic values related to sampling sites' geographic locations. 255 free-ranging bottlenose dolphins. Data from samples collected during multiple bottlenose dolphin capture-release projects conducted at 4 southeastern US coastal locations in 2000 through 2006 were combined to determine reference intervals for 52 clinicopathologic variables. A nonparametric bootstrap approach was applied to estimate 95th percentiles and associated 90% confidence intervals; the need for partitioning by length and sex classes was determined by testing for differences in estimated thresholds with a bootstrap method. When appropriate, quantile regression was used to determine continuous functions for 95th percentiles dependent on length. The proportion of out-of-range samples for all clinicopathologic measurements was examined for each geographic site, and multivariate ANOVA was applied to further explore variation in leukocyte subgroups. A need for partitioning by length and sex classes was indicated for many clinicopathologic variables. For each geographic site, few significant deviations from expected number of out-of-range samples were detected. Although mean leukocyte counts did not vary among sites, differences in the mean counts for leukocyte subgroups were identified. Although differences in the centrality of distributions for some variables were detected, the 95th percentiles estimated from the pooled data were robust and applicable across geographic sites. The derived reference intervals provide critical information for conducting bottlenose dolphin population health studies.


Exercise-induced bronchospasm in the elite athlete.
The term exercise-induced bronchospasm (EIB) describes the acute transient airway narrowing that occurs during and most often after exercise in 10 to 50% of elite athletes, depending upon the sport examined. Although multiple factors are unquestionably involved in the EIB response, airway drying caused by a high exercise-ventilation rate is primary in most cases. The severity of this reaction reflects the allergic predisposition of the athlete, the water content of the inspired air, the type and concentration of air pollutants inspired, and the intensity (or ventilation rate) of the exercise. The highest prevalence of EIB is seen in winter-sport populations, where athletes are chronically exposed to cold dry air and/or environmental pollutants found in indoor ice arenas. When airway surface liquid lost during the natural warming and humidification process of respiration is not replenished at a rate equal to the loss, the ensuing osmolarity change stimulates the release of inflammatory mediators and results in bronchospasm; this cascade of events is exacerbated by airway inflammation and airway remodelling. The acute EIB response is characterised by airway smooth muscle contraction, membrane swelling, and/or mucus plug formation. Evidence suggests that histamine, leukotrienes and prostanoids are likely mediators for this response. Although the presence of symptoms and a basic physical examination are marginally effective, objective measures of lung function should be used for accurate and reliable diagnosis of EIB. Diagnosis should include baseline spirometry, followed by an appropriate bronchial provocation test. To date, the best test to confirm EIB may simply be standard pulmonary function testing before and after high-intensity dry air exercise. A 10% post-challenge fall in forced expiratory volume in 1 second is used as diagnostic criteria. The goal of medical intervention is to limit EIB exacerbation and allow the athlete to train and compete symptom free. This is attempted through daily controller medications such as inhaled corticosteroids or by the prophylactic use of medications before exercise. In many cases, EIB is difficult to control. These and other data suggest that EIB in the elite athlete is in contrast with classic asthma.


The alveolate translation initiation factor 4E family reveals a custom toolkit for translational control in core dinoflagellates.
Dinoflagellates are eukaryotes with unusual cell biology and appear to rely on translational rather than transcriptional control of gene expression. The eukaryotic translation initiation factor 4E (eIF4E) plays an important role in regulating gene expression because eIF4E binding to the mRNA cap is a control point for translation. eIF4E is part of an extended, eukaryote-specific family with different members having specific functions, based on studies of model organisms. Dinoflagellate eIF4E diversity could provide a mechanism for dinoflagellates to regulate gene expression in a post-transcriptional manner. Accordingly, eIF4E family members from eleven core dinoflagellate transcriptomes were surveyed to determine the diversity and phylogeny of the eIF4E family in dinoflagellates and related lineages including apicomplexans, ciliates and heterokonts. The survey uncovered eight to fifteen (on average eleven) different eIF4E family members in each core dinoflagellate species. The eIF4E family members from heterokonts and dinoflagellates segregated into three clades, suggesting at least three eIF4E cognates were present in their common ancestor. However, these three clades are distinct from the three previously described eIF4E classes, reflecting diverse approaches to a central eukaryotic function. Heterokonts contain four clades, ciliates two and apicomplexans only a single recognizable eIF4E clade. In the core dinoflagellates, the three clades were further divided into nine sub-clades based on the phylogenetic analysis and species representation. Six of the sub-clades included at least one member from all eleven core dinoflagellate species, suggesting duplication in their shared ancestor. Conservation within sub-clades varied, suggesting different selection pressures. Phylogenetic analysis of eIF4E in core dinoflagellates revealed complex layering of duplication and conservation when compared to other eukaryotes. Our results suggest that the diverse eIF4E family in core dinoflagellates may provide a toolkit to enable selective translation as a strategy for controlling gene expression in these enigmatic eukaryotes.


Rapid colorimetric lactoferrin-based sandwich immunoassay on cotton swabs for the detection of foodborne pathogenic bacteria.
Cotton swab is the conventional swabbing tool that is usually applied for collecting pathogens from contaminated surfaces, followed by cells lysis and DNA extraction before subjecting to genetic analysis. However, such an approach is time consuming as it involves several steps and requires highly trained personnel to perform the experiment. In this study, we developed a new cotton swab-based detection system that involved integrating bacterial collection, preconcentration and detection on Q-tips. The platform is based on a sandwich assay that can detect different pathogens visually by color changes. Lactoferrin-immobilized cotton is used as a general capturing tool to collect various pathogens from surfaces. The presence of particular bacteria is then detected by immersing the cotton in antibodies attached to different colored nanobeads. The target cell is captured between the lactoferrin and specific antibody-conjugated beads which results in certain color development. The effectiveness of this simply fabricated sensor was demonstrated using Salmonella typhimurium, Salmonella enteritidis, Staphylococcus aureus and Campylobacter jejuni. The intensity of the color on the cotton surfaces increased with increasing the concentration of the pathogenic bacteria. The detection limit was as low as 10 cfu/ml for Salmonella typhimurium and Campylobacter jejuni, 100 cfu/ml for Salmonella enteritidis and 100 cfu/ml for Staphylococcus aureus on chicken meat surface. Moreover, this method showed high selectivity and was further confirmed by loop-mediated isothermal amplification (LAMP). The simplicity and the low cost of this colorimetric sensor renders it applicable to a wide range of other pathogens on different surfaces.


Transmission dynamics of heritable silencing induced by double-stranded RNA in Caenorhabditis elegans.
Heritable silencing effects are gene suppression phenomena that can persist for generations after induction. In the majority of RNAi experiments conducted in Caenorhabditis elegans, the silencing response results in a hypomorphic phenotype where the effects recede after the F1 generation. F2 and subsequent generations revert to the original phenotype. Specific examples of transgenerational RNAi in which effects persist to the F2 generation and beyond have been described. In this study, we describe a systematic pedigree-based analysis of heritable silencing processes resulting from initiation of interference targeted at the C. elegans oocyte maturation factor oma-1. Heritable silencing of oma-1 is a dose-dependent process where the inheritance of the silencing factor is unequally distributed among the population. Heritability is not constant over generational time, with silenced populations appearing to undergo a bottleneck three to four generations following microinjection of RNA. Transmission of silencing through these generations can be through either maternal or paternal gamete lines and is surprisingly more effective through the male gametic line. Genetic linkage tests reveal that silencing in the early generations is transmitted independently of the original targeted locus, in a manner indicative of a diffusible epigenetic element.


Evaluation of serum protein-based arrival formula and serum protein supplement (Gammulin) on growth, morbidity, and mortality of stressed (transport and cold) male dairy calves.
Previous studies with calves and other species have provided evidence that blood serum-derived proteins and fructooligosaccharides (FOS) may benefit intestinal health. We assessed the effects of supplementing products containing serum proteins as a component of arrival fluid support or serum proteins plus FOS (in addition to additional solids, minerals, and vitamins) in an early life dietary supplement on performance, morbidity, and mortality of stressed (transport, cold) male calves. Male Holstein calves (n=93) <1 wk old were stratified by arrival body weight (BW) and plasma protein concentration, and then randomly assigned to 1 of 4 treatment groups in a 2×2 factorial arrangement of one-time administration of fluid support [either control electrolyte solution (E) or the serum protein-containing arrival formula (AF)] and 14d of either no supplementation (NG) or supplementation with Gammulin (G; APC Inc., Ankeny, IA), which contains serum proteins and FOS in addition to other solids, minerals, and vitamins. Upon arrival at the research facility, calves were orally administered either AF or E. At the next feeding, half of the calves from each fluid support treatment received either milk replacer (20% crude protein, 20% fat) or the same milk replacer supplemented with G (50g/d during the first 14d). Starter and water were freely available. Feed offered and refused was recorded daily. Calf health was assessed by daily assignment of fecal and respiratory scores. Stature measures and BW were determined weekly. Blood samples were obtained at d 0 (before treatments), 2, 7, 14, and 28. Calves were weaned at d 42 and remained in the experiment until d 56. After 2 wk of treatments, calves previously fed AF had greater body length (66.6 vs. 66.0cm), intakes of dry matter (38.7 vs. 23.5g/d) and crude protein (9.2 vs. 5.6g/d) from starter, and cortisol concentration in blood (17.0 vs. 13.9 ng/mL) than calves fed E. Supplementation with G resulted in greater BW gain during the first 2 wk, increased intakes of dry matter and CP, and decreased respiratory scores. For the 8-wk experiment, G supplementation resulted in lower mean fecal score (1.6 vs. 1.8) and fewer antibiotic treatments per calf (1.5 vs. 2.5) than NG. Survival was greater in G than in NG calves (98 vs. 84%). Despite the marked reduction in morbidity and mortality, blood indicators of acute-phase response, urea N, and total protein were not affected by AF or G in transported cold-stressed male calves.


Reduced RNA expression of the FMR1 gene in women with low (CGGn<26) repeats.
Low FMR1 variants (CGGn<26) have been associated with premature ovarian aging, female infertility and poor IVF treatment success. Until now, there is little published information concerning possible molecular mechanisms for this effect. We wished to examine whether relative expression of RNA and the FMR1 gene's fragile X mental retardation protein (FMRP) RNA isoforms differ in women with various FMR1 sub-genotypes (normal, low CGGn<26 and/or high CGGn≥34). This prospective cohort study was conducted between 2014 and 2017 in a clinical research unit of the Center for Human Reproduction in New York City. The study involved a total of 98 study subjects, including 18 young oocyte donors and 80 older infertility patients undergoing routine in vitro fertilization (IVF) cycles. The main outcome measure was RNA expression in human luteinized granulosa cells of 5 groups of FMRP isoforms. The relative expression of FMR1 RNA in human luteinized granulosa cells was measured by real-time PCR and a possible association with CGGn was explored. All 5 groups of FMRP RNA isoforms examined were found to be differentially expressed in human luteinized granulosa cells. The relative expression of four FMR1 RNA isoforms showed significant differences among 6 FMR1 sub-genotypes. Women with at least one low allele expressed significantly lower levels of all 5 sets of FRMP isoforms in comparison to the non-low group. While it would be of interest to see whether FMRP is also decreased in the low-group we recognize that in recent years it has been increasingly documented that information flow of genetics may be regulated by non-coding RNA, that is, without translation to a protein product. We, thus, conclude that various CGG expansions of FMR1 allele may lead to changes of RNA levels and ratios of distinct FMRP RNA isoforms, which could regulate the translation and/or cellular localization of FMRP, affect the expression of steroidogenic enzymes and hormonal receptors, or act in some other epigenetic process and therefore result in the ovarian dysfunction in infertility.


The effects of combined micron-/submicron-scale surface roughness and nanoscale features on cell proliferation and differentiation.
Titanium (Ti) osseointegration is critical for the success of dental and orthopedic implants. Previous studies have shown that surface roughness at the micro- and submicro-scales promotes osseointegration by enhancing osteoblast differentiation and local factor production. Only relatively recently have the effects of nanoscale roughness on cell response been considered. The aim of the present study was to develop a simple and scalable surface modification treatment that introduces nanoscale features to the surfaces of Ti substrates without greatly affecting other surface features, and to determine the effects of such superimposed nano-features on the differentiation and local factor production of osteoblasts. A simple oxidation treatment was developed for generating controlled nanoscale topographies on Ti surfaces, while retaining the starting micro-/submicro-scale roughness. Such nano-modified surfaces also possessed similar elemental compositions, and exhibited similar contact angles, as the original surfaces, but possessed a different surface crystal structure. MG63 cells were seeded on machined (PT), nano-modified PT (NMPT), sandblasted/acid-etched (SLA), and nano-modified SLA (NMSLA) Ti disks. The results suggested that the introduction of such nanoscale structures in combination with micro-/submicro-scale roughness improves osteoblast differentiation and local factor production, which, in turn, indicates the potential for improved implant osseointegration in vivo.


Screening for breast cancer: the Swedish trial.
A randomized, controlled mammographic screening procedure has been initiated in Sweden. The study has been conducted since 1977 in Kopparberg county and since 1978 in Ostergötland county. This paper describes the planning and organization of the study as well as the method of screening used. The results of the first 27 months in Kopparberg county are presented.


Risk behaviors of 15-21 year olds in Mexico lead to a high prevalence of sexually transmitted infections: results of a survey in disadvantaged urban areas.
Due to the fact that adolescents are more likely to participate in high-risk behaviors, this sector of the population is particularly vulnerable to contracting sexually transmitted infections (STIs) and resultant health problems. A survey was carried out among adolescents from poor homes in 204 small-urban areas of Mexico. Information was collected in relation to risk behaviors and socio-economic environment. A sub-group of the participants also provided blood and urine samples which were analyzed to detect sexually transmitted infections. The presence of Chlamydia was detected in nearly 8% of participants who had stated that they were sexually active (18%) and approximately 12% were positive for herpes type 2-specific antibodies. For both, a greater proportion of girls resulted positive compared to boys. The presence of these biological outcomes of sexual risk behavior was associated with other risk behaviors (smoking), but not with self-reported indicators of protected sex (reported use of condom during most recent sexual activity). The results presented in this study show a startlingly high prevalence of HSV-2 among sexually active Mexican adolescents in poor urban areas, suggesting that this group has participated to a great extent in risky sexual practices. The relationships between socioeconomic environment and adolescent risk behavior need to be better understood if we are to design preventive interventions that modify the determinants of risk behaviors.


Reaction to intravenous contrast media mimicking intracerebral hemorrhage after percutaneous coronary intervention.
Cerebroavascular complications after percutaneous coronary interventions, although rare, need immediate diagnostic and treatment in order to minimize the possible sequelae. At the end of a percutaneous coronary intervention a 63-year old man, long standing migraineur, developed severe headache, confusion, nausea, photophobia, slow speech and left hemiparesis. In the presence of hyperdensity of the right parietal area in the brain CT, he was suspected to suffer from intracerebral hemorrhage, and treatment with platelet glycoprotein IIb/IIIa inhibitors was immediately stopped. Revision of the CT and disappearance of the clinical and radiological signs were concordant with the diagnosis of intraparenchymal extravasation of the contrast media. We report a very rare complication of contrast media after coronary angiography with extravasation unilaterally in the anterior circulation. Recognition of this complication can avoid unnecessary investigations.


The relationship between primary health care practitioners' job satisfaction and characteristics of their practice settings.
Today, individuals spend much of their time at the workplace, and theorists argue that an individual's employment experience is an integral part of his or her social identity. Current research emphasizes the importance of job satisfaction to a person's entire life adjustment. Yet little data exists about the job satisfaction of nurses working in the advanced practice role of primary care. The purposes of this article's study were to describe those individual and situational factors that contribute to job satisfaction and dissatisfaction among nurse practitioners, and to explore the relationship between levels of job satisfaction and characteristics of primary health care practitioners' work/practice settings. The study results suggest that, overall, NPs demonstrate a high level of job satisfaction; only 9 percent reported overall dissatisfaction with their jobs. Autonomy, sense of accomplishment and time spent in patient care ranked as the top three factors contributing to job satisfaction. The highest ranked dissatisfier was salary. Differences in satisfaction existed according to several practice-setting characteristics. Fewer factors than initially expected were associated with overall job satisfaction. The correlates that were noted, however, identified number of years of experience and self-perceived skill levels as important variables contributing to overall job satisfaction. Implications of these findings for the early guidance of practitioners assuming the advanced-practice role and for the NP educational curriculum are discussed.


Comparison of an inertial sensor system with a stationary force plate for evaluation of horses with bilateral forelimb lameness.
To assess the analytic sensitivity of an inertial sensor system for detection of the more severely affected forelimb in horses with bilateral lameness. 18 adult horses with forelimb lameness. Horses were fitted with inertial sensors and evaluated for lameness with a stationary force plate as they were trotted in a straight line. Inertial sensor-derived measurements for vertical head movement asymmetry (HMA) and vector sum (VS) of maximum and minimum head height differences between right and left halves of the stride were used to predict differences in mean peak vertical force (PVF) as a percentage of body weight between the right and left forelimbs. Repeatability was compared by calculation of the intraclass correlation coefficient (ICC) for each variable. Correct classification percentages for the lamer forelimb were determined by use of a stationary force plate as the standard. SEs of the prediction of difference in PVF between the right and left forelimbs from HMA and VS were 6.1% and 5.2%, respectively. Head movement asymmetry (ICC, 0.72) was less repeatable than PVF (ICC, 0.86) and VS (ICC, 0.84). Associations were positive and significant between HMA (R(2) = 0.73) and VS (R(2) = 0.81) and the difference in PVF between the right and left forelimbs. Correct classification percentages for HMA and VS for detecting the lamer forelimb were 83.3% and 77.8%, respectively. Results suggested that an inertial sensor system to measure vertical asymmetry (HMA and VS) due to forelimb lameness in horses trotting in a straight line has adequate analytic sensitivity for clinical use. Additional studies are required to assess specificity of the system.


The reticulons: a family of proteins with diverse functions.
The reticulon family is a large and diverse group of membrane-associated proteins found throughout the eukaryotic kingdom. All of its members contain a carboxy-terminal reticulon homology domain that consists of two hydrophobic regions flanking a hydrophilic loop of 60-70 amino acids, but reticulon amino-terminal domains display little or no similarity to each other. Reticulons principally localize to the endoplasmic reticulum, and there is evidence that they influence endoplasmic reticulum-Golgi trafficking, vesicle formation and membrane morphogenesis. However, mammalian reticulons have also been found on the cell surface and mammalian reticulon 4 expressed on the surface of oligodendrocytes is an inhibitor of axon growth both in culture and in vivo. There is also growing evidence that reticulons may be important in neurodegenerative diseases such as Alzheimer's disease and amyotrophic lateral sclerosis. The diversity of structure, topology, localization and expression patterns of reticulons is reflected in their multiple, diverse functions in the cell.


Cryptococcus neoformans Induces MCP-1 Release and Delays the Death of Human Mast Cells.
Cryptococcosis, caused by the basidiomycete Cryptococcus neoformans, is a life-threatening disease affecting approximately one million people per year worldwide. Infection can occur when C. neoformans cells are inhaled by immunocompromised people. In order to establish infection, the yeast must bypass recognition and clearance by immune cells guarding the tissue. Using in vitro infections, we characterized the role of mast cells (MCs) in cryptococcosis. We found that MCs recognize C. neoformans and release inflammatory mediators such as tryptase and cytokines. From the latter group MCs released mainly CCL-2/MCP-1, a strong chemoattractant for monocytic cells. We demonstrated that supernatants of infected MCs recruit monocytes but not neutrophils. During infection with C. neoformans, MCs have a limited ability to kill the yeast depending on the serotype. C. neoformans, in turn, modulates the lifespan of MCs both, by presence of its polysaccharide capsule and by secreting soluble modulators. Taken together, MCs might have important contributions to fungal clearance during early stages of cryptocococis where these cells regulate recruitment of monocytes to mucosal tissues.


Target-specific requirements for enhancers of decapping in miRNA-mediated gene silencing.
microRNAs (miRNAs) silence gene expression by suppressing protein production and/or by promoting mRNA decay. To elucidate how silencing is accomplished, we screened an RNA interference library for suppressors of miRNA-mediated regulation in Drosophila melanogaster cells. In addition to proteins known to be required for miRNA biogenesis and function (i.e., Drosha, Pasha, Dicer-1, AGO1, and GW182), the screen identified the decapping activator Ge-1 as being required for silencing by miRNAs. Depleting Ge-1 alone and/or in combination with other decapping activators (e.g., DCP1, EDC3, HPat, or Me31B) suppresses silencing of several miRNA targets, indicating that miRNAs elicit mRNA decapping. A comparison of gene expression profiles in cells depleted of AGO1 or of individual decapping activators shows that approximately 15% of AGO1-targets are also regulated by Ge-1, DCP1, and HPat, whereas 5% are dependent on EDC3 and LSm1-7. These percentages are underestimated because decapping activators are partially redundant. Furthermore, in the absence of active translation, some miRNA targets are stabilized, whereas others continue to be degraded in a miRNA-dependent manner. These findings suggest that miRNAs mediate post-transcriptional gene silencing by more than one mechanism.


Evidence against anandamide as the hyperpolarizing factor mediating the nitric oxide-independent coronary vasodilator effect of bradykinin in the rat.
The mediator of nitric oxide-(NO) independent vasodilation attributed to endothelium-derived hyperpolarizing factor remains unidentified although there is evidence for a cytochrome P450-derived eicosanoid. Anandamide, the ethanolamide of arachidonic acid and an endogenous ligand for cannabinoid receptors, was proposed as an endothelium-derived hyperpolarizing factor-mediating mesenteric vasodilation to acetylcholine and the hypotensive effect of bradykinin. Using pharmacological interventions that attenuate responses to bradykinin, we examined the possibility of anandamide as a mediator of the NO-independent vasodilator effect of bradykinin in the rat perfused heart by determining responses to anandamide and arachidonic acid. Hearts were treated with indomethacin to exclude prostaglandins and nitroarginine to inhibit NO synthesis and elevate perfusion pressure. The cannabinoid receptor antagonist, SR 141716A (2 microM), reduced dose-dependent vasodilator responses to anandamide (1-10 microgram) but was without effect on responses to AA (1-10 microgram), bradykinin (10-1000 ng) or cromakalim (1-10 microgram). Inhibition of voltage-dependent Ca++ channels with nifedipine (5 nM) attenuated vasodilation to anandamide and arachidonic acid whereas inhibition of Ca++-activated K+ channels with charybdotoxin (10 nM) reduced responses to arachidonic acid but had no effect on vasodilation induced by anandamide. Inhibition of cytochrome P450 with clotrimazole (1 microM) greatly reduced vasodilator responses to bradykinin with less effect on those to anandamide. Finally, the time course of the coronary vasodilator responses to anandamide and bradykinin were dissimilar. These results argue against a role of anandamide in the vasodilator effect of bradykinin in the rat heart.


Biomimicking Fiber Scaffold as an Effective In Vitro and In Vivo MicroRNA Screening Platform for Directing Tissue Regeneration.
MicroRNAs effectively modulate protein expression and cellular response. Unfortunately, the lack of robust nonviral delivery platforms has limited the therapeutic application of microRNAs. Additionally, there is a shortage of drug-screening platforms that are directly translatable from in vitro to in vivo. Here, a fiber substrate that provides nonviral delivery of microRNAs for in vitro and in vivo microRNA screening is introduced. As a proof of concept, difficult-to-transfect primary neurons are targeted and the efficacy of this system is evaluated in a rat spinal cord injury model. With this platform, enhanced gene-silencing is achieved in neurons as compared to conventional bolus delivery (p < 0.05). Thereafter, four well-recognized microRNAs (miR-21, miR-222, miR-132, and miR-431) and their cocktails are screened systematically. Regardless of age and origin of the neurons, similar trends are observed. Next, this fiber substrate is translated into a 3D system for direct in vivo microRNA screening. Robust nerve ingrowth is observed as early as two weeks after scaffold implantation. Nerve regeneration in response to the microRNA cocktails is similar to in vitro experiments. Altogether, the potential of the fiber platform is demonstrated in providing effective microRNA screening and direct translation into in vivo applications.


Phylogenetic Analysis of Bean yellow mosaic virus Isolates from Four Continents: Relationship Between the Seven Groups Found and Their Hosts and Origins.
Genetic diversity of Bean yellow mosaic virus (BYMV) was studied by comparing sequences from the coat protein (CP) and genome-linked viral protein (VPg) genes of isolates from four continents. CP sequences compared were those of 17 new isolates and 47 others already on the database, while the VPg sequences used were from four new isolates and 10 from the database. Phylogenetic analysis of the CP sequences revealed seven distinct groups, six polytypic and one monotypic. The largest and most genetically diverse polytypic group, which had intragroup diversity of 0.061 nucleotide substitutions per site, contained isolates from natural infections in eight host species. These original isolation hosts included both wild (four) and domesticated (four) species and were from monocotyledonous and dicotyledonous plant families, indicating a generalized natural host range strategy. Only one of the other five polytypic groups spanned both monocotyledons and dicotyledons, and all contained isolates from fewer species (one to four), all of which were domesticated and had lower intragroup diversity (0.019 to 0.045 nucleotide substitutions per site), indicating host specialization. Phylogenetic analysis of the fewer VPg sequences revealed three polytypic and two monotypic groupings. These groups also correlated with original natural isolation hosts, but the branch topologies were sometimes incongruous with those formed by CPs. Also, intragroup diversity was generally higher for VPgs than for CPs. A plausible explanation for the groups found when the 64 different CP sequences were compared is that the generalized group represents the original ancestral type from which the specialist host groups evolved in response to domestication of plants after the advent of agriculture. Data on the geographical origins of the isolates within each group did not reveal whether the specialized groups might have coevolved with their principal natural hosts where these were first domesticated, but this seems plausible.


Transcriptional and post-transcriptional regulation of tyrosine hydroxylase messenger RNA in PC12 cells during persistent stimulation by VIP and PACAP38: differential regulation by protein kinase A and protein kinase C-dependent pathways.
VIP and PACAP38 are closely related peptides that are released in the adrenal gland and sympathetic ganglia and regulate catecholamine synthesis and release. We used PC12 cells as a model system to examine receptor and second messenger pathways by which each peptide stimulates transcriptional and post-transcriptional mechanisms that regulate the level of the mRNA for tyrosine hydroxylase (TH), the rate-limiting enzymatic step in catecholamine synthesis. Concentration-response studies revealed that PACAP38 had both greater efficacy and potency than VIP. The specific PAC1 receptor antagonist PACAP[6-38] blocked the effects of each peptide on TH mRNA content while the PACAP/VIP type II receptor antagonist (N-AC-Tyr(1)-D-Phe(2))-GRF-(1-29)-NH(2) was without effect. At equipotent concentrations, each peptide stimulated a transient increase in TH gene transcription lasting less than 3h. Continuous VIP treatment stimulated a transient increase in TH mRNA lasting less than 24h. In contrast, continuous exposure to PACAP38 stimulated a stable increase in TH mRNA that persisted for 2 days in the absence of elevated transcription, pointing to different post-transcriptional effects of the two peptides. PACAP38 alone had no effect on the magnitude of TH gene transcription or TH mRNA in A126-1B2 PKA-deficient PC12 cells. However, when combined with dexamethasone, PACAP38 produced a synergistic increase in TH mRNA in the absence of PACAP38-stimulated TH gene transcription. In contrast, VIP had no effect on either TH mRNA content or TH gene transcription in this model. PACAP38, but not VIP, stimulated PKC activity. Calphostin C antagonized the effect of PACAP38 on the persistent post-transcriptional elevation in TH mRNA. Thus, the results support the conclusion that VIP and PACAP38 each stimulate PAC1 receptors to increase TH gene transcription through a PKA-controlled pathway, but their divergent post-transcriptional effects result at least partly from differing abilities to stimulate PKC.


Physical symptoms 14 months after a natural disaster in individuals with or without injury are associated with different types of exposure.
To investigate whether different types of exposure to the 2004 tsunami were associated with physical symptoms 14 months after the disaster and to study correlations between survivors' physical and psychological symptoms. Using a cross-sectional design, 1505 survivors from the 2004 Indian Ocean Tsunami, tourists from Stockholm, who had been present in the disaster areas, responded to a postal questionnaire. Eight groups based on type of exposure were created. Physical symptoms occurring on a daily or weekly basis over the past year were investigated in four indices: musculoskeletal, cardiorespiratory, neurological, and gastrointestinal. Mental health symptoms (General Health Questionnaire-12) and posttraumatic stress symptoms (Impact of Event Scale-Revised) were also investigated. Multiple logistic regression analyses were conducted with controls for background variables and exposure, with physical symptoms as outcome variables. The association between physical and psychological symptoms was studied with the Spearman Rank Order Correlation. Different types of exposure during the disaster were associated with physical symptoms 14 months later for survivors both with and without severe physical injury. The single exposure of life threat, also in combination with other exposures, was associated with a higher risk for reporting of physical symptoms. Physical symptoms showed modest yet significant correlation with psychological symptoms. It is important to pay attention to both physical and psychological symptoms among disaster survivors whether they have been injured or not. A relatively simple questionnaire about physical symptoms may be a good complement to the scales used to assess psychological problems after disaster.


The timing and role of initiation rites in preparing young people for adolescence and responsible sexual and reproductive behaviour in Malawi.
This paper examines timing of puberty and mechanisms through which society prepares adolescents to understand and deal with it in Malawi. Data from a national representative survey of adolescents and in-depth interviews also conducted with adolescents are used. SPSS was used to analyse survey data while N6 was used to analyse qualitative data. Results show that the onset of menarche in girls and various pubertal body changes in boys can be a cause of joy, excitement, or distress depending on how adolescents understand what this means to them at this critical stage when they start defining and comprehending their sexuality. Much more emphasis is put on educating girls about reproductive implications of menarche than on what is expected of boys as sexual beings, which may contribute to boys' greater indulgence in risky sexual behaviors than girls. The significance of initiation ceremonies in some communities provides an important platform through which programs can reach many adolescents and intervene, particularly in addressing the widely held notion among initiates that attending these ceremonies symbolizes that one is not a child anymore and can have sex.


Determination of total iodine in infant formula and nutritional products by inductively coupled plasma/mass spectrometry: single-laboratory validation.
A single-laboratory validation by inductively coupled plasma-mass spectrometry was developed for the determination total iodine (m/z 127) in infant formula and adult nutritional products. All samples were digested in nitric acid using a closed vessel microwave oven system; Te (m/z 130) was used as an internal standard. To prevent loss of iodine, ammonium hydroxide solution was added to the samples immediately after digestion. The method quantitation limit for total iodine was 0.3 ng/mL, but a practical LOQ was used at 1.0 ng/mL, a concentration at which there was a negligible bias due to nonlinearity. The total iodine concentrations (112-1900 ng/g) in 14 out of 15 nutritional products were within specification limits. Within-day and day-to-day (6 independent days) precision values were < 10% RSD. The observed precision for the overall mean (18 independent days) of a control sample was approximately 4% RSD. In two National Institute of Standards and Technology standard reference materials, total iodine results were within certified limits. Sample spike recoveries for all 15 nutritional products were 92-105%. The data show that a conventional microwave oven digestion procedure can be used to prepare samples for iodine determination. Therefore, this technique is very compatible with other methods being proposed as modern official methods for the analysis of minerals in nutritional products.


A collection of genetically engineered Populus trees reveals wood biomass traits that predict glucose yield from enzymatic hydrolysis.
Wood represents a promising source of sugars to produce bio-based renewables, including biofuels. However, breaking down lignocellulose requires costly pretreatments because lignocellulose is recalcitrant to enzymatic saccharification. Increasing saccharification potential would greatly contribute to make wood a competitive alternative to petroleum, but this requires improving wood properties. To identify wood biomass traits associated with saccharification, we analyzed a total of 65 traits related to wood chemistry, anatomy and structure, biomass production and saccharification in 40 genetically engineered Populus tree lines. These lines exhibited broad variation in quantitative traits, allowing for multivariate analyses and mathematical modeling. Modeling revealed that seven wood biomass traits associated in a predictive manner with saccharification of glucose after pretreatment. Four of these seven traits were also negatively associated with biomass production, suggesting a trade-off between saccharification potential and total biomass, which has previously been observed to offset the overall sugar yield from whole trees. We therefore estimated the "total-wood glucose yield" (TWG) from whole trees and found 22 biomass traits predictive of TWG after pretreatment. Both saccharification and TWG were associated with low abundant, often overlooked matrix polysaccharides such as arabinose and rhamnose which possibly represent new markers for improved Populus feedstocks.


Functional dissection of the ARGONAUTE7 promoter.
ARGONAUTES are the central effector proteins of RNA silencing which bind target transcripts in a small RNA-guided manner. Arabidopsis thaliana has 10 ARGONAUTE (AGO) genes, with specialized roles in RNA-directed DNA methylation, post-transcriptional gene silencing, and antiviral defense. To better understand specialization among AGO genes at the level of transcriptional regulation we tested a library of 1497 transcription factors for binding to the promoters of AGO1,AGO10, and AGO7 using yeast 1-hybrid assays. A ranked list of candidate DNA-binding TFs revealed binding of the AGO7 promoter by a number of proteins in two families: the miR156-regulated SPL family and the miR319-regulated TCP family, both of which have roles in developmental timing and leaf morphology. Possible functions for SPL and TCP binding are unclear: we showed that these binding sites are not required for the polar expression pattern of AGO7, nor for the function of AGO7 in leaf shape. Normal AGO7 transcription levels and function appear to depend instead on an adjacent 124-bp region. Progress in understanding the structure of this promoter may aid efforts to understand how the conserved AGO7-triggered TAS3 pathway functions in timing and polarity.


Translational research in pediatrics II: blood collection, processing, shipping, and storage.
Translational research often involves tissue sampling and analysis. Blood is by far the most common tissue collected. Due to the many difficulties encountered with blood procurement from children, it is imperative to maximize the quality and stability of the collected samples to optimize research results. Collected blood can remain whole or be fractionated into serum, plasma, or cell concentrates such as red blood cells, leukocytes, or platelets. Serum and plasma can be used for analyte studies, including proteins, lipids, and small molecules, and as a source of cell-free nucleic acids. Cell concentrates are used in functional studies, flow cytometry, culture experiments, or as a source for cellular nucleic acids. Before initiating studies on blood, a thorough evaluation of practices that may influence analyte and/or cellular integrity is required. Thus, it is imperative that child health researchers working with human blood are aware of how experimental results can be altered by blood sampling methods, times to processing, container tubes, presence or absence of additives, shipping and storage variables, and freeze-thaw cycles. The authors of this review, in an effort to encourage and optimize translational research using blood from pediatric patients, outline best practices for blood collection, processing, shipment, and storage.


SMO algorithm for least-squares SVM formulations.
This article extends the well-known SMO algorithm of support vector machines (SVMs) to least-squares SVM formulations that include LS-SVM classification, kernel ridge regression, and a particular form of regularized kernel Fisher discriminant. The algorithm is shown to be asymptotically convergent. It is also extremely easy to implement. Computational experiments show that the algorithm is fast and scales efficiently (quadratically) as a function of the number of examples.


Temporal and local appearance of alkaline phosphatase activity in early stages of guided bone regeneration. A descriptive histochemical study in humans.
Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters and it seems to be a prerequisite for normal skeletal mineralization. Also, ALP is the most widely recognized marker of osteoblast phenotypes. By a tissue regenerative technique called Guided Bone Regeneration (GBR), it is possible nowadays to regenerate small bony defects. The aim of the present study was to investigate early events in bone healing and neogenesis by studying histochemically the temporal and local appearance of the marker Alkaline Phosphatase (ALP) in a GBR model system. Nine healthy volunteers (5 males, 4 females, mean age 31.7 years) participated in the experiment. After raising a mucoperiosteal flap from the mandibular second molar to the retromolar area in each volunteer, a hollow titanium test cylinder was placed into a congruent bony bed and the coronal end of the cylinder was closed with an ePTFE-membrane. Then the flap was adapted and sutured to obtain primary wound closure. After 2, 7 and 12 weeks, the regenerated tissue within the cylinders was harvested. Histologically, ALP activity was observed associated with the osteoid seams in the very basal part of the regenerate where new bone trabeculae were in the process of being formed. More coronally, large round cells seemed to secrete an ALP-positive substance since in the center of such cell clusters strong ALP activity located extracellularly was detected. In the present experiment, ALP seemed to have been an early sign of osteoblast secretion of a matrix which subsequently was determined to become osteoid. ALP activity was never seen isolated within connective tissue and away from bone. This is an indication that its source is linked to existing bone. The present study has documented for the first time the appearance of ALP activity in guided bone regenerations in humans. It has revealed that: 1) Osteogenesis in guided bone regeneration is preceded by localized, marked expression of ALP in an organized connective tissue environment. 2) Bone neogenesis is an early event in this experimental setup and may be detected already 2 weeks after wounding. 3) Expression of ALP and subsequent bone neogenesis is originating from and topographically linked to pre-existing bone structures.


Combined covalent-electrostatic model of hydrogen bonding improves structure prediction with Rosetta.
Interactions between polar atoms are challenging to model because at very short ranges they form hydrogen bonds (H-bonds) that are partially covalent in character and exhibit strong orientation preferences; at longer ranges the orientation preferences are lost, but significant electrostatic interactions between charged and partially charged atoms remain. To simultaneously model these two types of behavior, we refined an orientation dependent model of hydrogen bonds [Kortemme et al. J. Mol. Biol. 2003, 326, 1239] used by the molecular modeling program Rosetta and then combined it with a distance-dependent Coulomb model of electrostatics. The functional form of the H-bond potential is physically motivated and parameters are fit so that H-bond geometries that Rosetta generates closely resemble H-bond geometries in high-resolution crystal structures. The combined potentials improve performance in a variety of scientific benchmarks including decoy discrimination, side chain prediction, and native sequence recovery in protein design simulations and establishes a new standard energy function for Rosetta.


Kv1.1-containing channels are critical for temporal precision during spike initiation.
Low threshold, voltage-gated potassium currents (Ikl) are widely expressed in auditory neurons that can fire temporally precise action potentials (APs). In the medial nucleus of the trapezoid body (MNTB), channels containing the Kv1.1 subunit (encoded by the Kcna1 gene) underlie Ikl. Using pharmacology, genetics and whole cell patch-clamp recordings in mouse brain slices, we tested the role of Ikl in limiting AP latency-variability (jitter) in response to trains of single inputs at moderate to high stimulation rates. With dendrotoxin-K (DTX-K, a selective blocker of Kv1.1-containing channels), we blocked Ikl maximally (approximately 80% with 100 nM DTX-K) or partially (approximately 50% with 1-h incubation in 3 nM DTX-K). Ikl was similar in 3 nM DTX-K-treated cells and cells from Kcna1(-/-) mice, allowing a comparison of these two different methods of Ikl reduction. In response to current injection, Ikl reduction increased the temporal window for AP initiation and increased jitter in response to the smallest currents that were able to drive APs. While 100 nM DTX-K caused the largest increases, latency and jitter in Kcna1(-/-) cells and in 3 nM DTX-K-treated cells were similar to each other but increased compared with +/+. The near-phenocopy of the Kcna1(-/-) cells with 3 nM DTX-K shows that acute blockade of a subset of the Kv1.1-containing channels is functionally similar to the chronic elimination of all Kv1.1 subunits. During rapid stimulation (100-500 Hz), Ikl reduction increased jitter in response to both large and small inputs. These data show that Ikl is critical for maintaining AP temporal precision at physiologically relevant firing rates.


Sugar-containing beverage intake in toddlers and body composition up to age 6 years: the Generation R study.
Intake of sugar-containing beverages (SCBs) has been associated with higher body mass index (BMI) in childhood. The potential effect of SCB intake during infancy is unclear. We examined the association of SCB intake at 13 months with BMI development until 6 years and body composition at age 6 years. This study included 2371 Dutch children from a population-based prospective cohort study. SCB intake at 13 months was assessed using a Food Frequency Questionnaire with validation against 24-h recalls and was standardized for total energy. BMI was calculated from repeated weight and height measurements, and age- and sex-specific s.d. scores were calculated. Adiposity was measured using Dual-energy X-ray absorptiometry. In girls, higher SCB intake at 13 months was significantly associated with higher BMI at ages 2, 3, 4 and 6 years (at age 6 years BMI (s.d. score) increase 0.11 (95% confidence interval (CI) +0.00; 0.23), high versus low intake). We observed a tendency towards higher android/gynoid fat ratio in girls with high intake (s.d. increase 0.14 (95% CI -0.02; 0.29), versus low intake) but not with body fat percentage. In boys, there was no association with BMI or body composition, but boys with high SCB intake at 13 months were taller at age 6 years (s.d. increase 0.14 (95% CI +0.00; 0.27), versus low intake). Higher SCB intake at 13 months was associated with higher BMI up to age 6 years in girls but not in boys. Our results imply that the unfavorable effects of SCB intake start early in life and that dietary advice regarding limiting SCB intake should already be given early in life.


Control of B cell development by the histone H2A deubiquitinase MYSM1.
Epigenetic histone modifications play critical roles in the control of gene transcription. Recently, an increasing number of histone H2A deubiquitinases have been identified and characterized. However, the physiological functions for this entire group of histone H2A deubiquitinases remain unknown. In this study, we revealed that the histone H2A deubiquitinase MYSM1 plays an essential and intrinsic role in early B cell development. MYSM1 deficiency results in a block in early B cell commitment and a defect of B cell progenitors in expression of EBF1 and other B lymphoid genes. We further demonstrated that MYSM1 derepresses EBF1 transcription in B cell progenitors by orchestrating histone modifications and transcription factor recruitment to the EBF1 locus. Thus, this study not only uncovers the essential role for MYSM1 in gene transcription during early B cell development but also underscores the biological significance of reversible epigenetic histone H2A ubiquitination.


Pattern evoked potentials in awake rhesus monkeys.
The pattern evoked potential (EP) in man to checkerboard stimulation has been shown to consist of various components originating in different regions of the visual cortex. Surface recordings, however, cannot unambiguously localize the sources of these components; for precision, depth recordings seem to be indicated. Considering the close correspondence in cortical architecture to man, rhesus monkeys (Macaca mulatta) seem to be a suitable animal for such experiments. However, Padmos et al. demonstrated that in monkeys anesthetized by pentobarbital (Nembutal), no pattern EP as found in man could be recorded. The present experiments were carried out to investigate whether the lack of contrast-specific EPs in monkey can be attributed to the effects of anesthesia. Four rhesus monkeys were trained to fixate at a television screen on which checkerboard and bar patterns of various sizes could be presented. The results from this study demonstrate that in monkey as in man, pattern EPs can be obtained that can be distinguished from those evoked by luminance variations. Therefore the awake rhesus monkey seems to be a suitable experimental model in the search for the origin of the pattern EP.


The facial flow concept: An organic orofacial analysis-the vertical component.
Orofacial analysis has been used by dentists for many years. The process involves applying mathematical rules, geometric principles, and straight lines to create either parallel or perpendicular references based on the true horizon and/or natural head position. These reference lines guide treatment planning and smile design for restorative treatments to achieve harmony between the new smile and the face. The goal is to obtain harmony and not symmetry. Faces are asymmetrical entities and because of that cannot be analyzed using purely straight lines. In this article, a more natural, organic, and dynamic process of evaluation is presented to minimize errors and generate harmoniously balanced smiles instead of perfect, mathematical smiles.


Synthesis and Structure-Activity Relationships of Arylsulfonamides as AIMP2-DX2 Inhibitors for the Development of a Novel Anticancer Therapy.
AIMP2-DX2, a splicing variant of AIMP2, is up-regulated in lung cancer, possesses oncogenic activity, and results in tumorigenesis. Specifically inhibiting the interaction between AIMP2-DX2 and HSP70 to suppress AIMP2-DX2-dependent cancers with small molecules is considered a promising avenue for cancer therapeutics. Optimization of hit BC-DXI-04 (IC50 = 40.1 μM) provided new potent sulfonamide based AIMP2-DX2 inhibitors. Among these, BC-DXI-843 showed improved inhibition against AIMP2-DX2 (IC50 = 0.92 μM) with more than 100-fold selectivity over AIMP2 in a luciferase assay. Several binding assays indicated that this compound effectively induces cancer cell apoptosis by specifically interrupting the interaction between DX2 and HSP70, which leads to the degradation of DX2 via Siah1-mediated ubiquitination. More importantly, BC-DXI-843 demonstrated in vivo efficacy in a tumor xenograft mouse model (H460 cells) at a dosage of 50 mg/kg, suggesting it as a promising lead for development of novel therapeutics targeting AIMP2-DX2 in lung cancer.


Fbxo45 forms a novel ubiquitin ligase complex and is required for neuronal development.
Fbxo45 is an F-box protein that is restricted to the nervous system. Unlike other F-box proteins, Fbxo45 was found not to form an SCF complex as a result of an amino acid substitution in the consensus sequence for Cul1 binding. Proteomics analysis revealed that Fbxo45 specifically associates with PAM (protein associated with Myc), a RING finger-type ubiquitin ligase. Mice deficient in Fbxo45 were generated and found to die soon after birth as a result of respiratory distress. Fbxo45(-)(/)(-) embryos show abnormal innervation of the diaphragm, impaired synapse formation at neuromuscular junctions, and aberrant development of axon fiber tracts in the brain. Similar defects are also observed in mice lacking Phr1 (mouse ortholog of PAM), suggesting that Fbxo45 and Phr1 function in the same pathway. In addition, neuronal migration was impaired in Fbxo45(-)(/)(-) mice. These results suggest that Fbxo45 forms a novel Fbxo45-PAM ubiquitin ligase complex that plays an important role in neural development.


Exon skipping as a therapeutic strategy applied to an RYR1 mutation with pseudo-exon inclusion causing a severe core myopathy.
Central core disease is a myopathy often arising from mutations in the type 1 ryanodine receptor (RYR1) gene, encoding the sarcoplasmic reticulum calcium release channel RyR1. No treatment is currently available for this disease. We studied the pathological situation of a severely affected child with two recessive mutations, which resulted in a massive reduction in the amount of RyR1. The paternal mutation induced the inclusion of a new in-frame pseudo-exon in RyR1 mRNA that resulted in the insertion of additional amino acids leading to the instability of the protein. We hypothesized that skipping this additional exon would be sufficient to restore RyR1 expression and to normalize calcium releases. We therefore developed U7-AON lentiviral vectors to force exon skipping on affected primary muscle cells. The efficiency of the exon skipping was evaluated at the mRNA level, at the protein level, and at the functional level using calcium imaging. In these affected cells, we observed a decreased inclusion of the pseudo-exon, an increased RyR1 protein expression, and a restoration of calcium releases of normal amplitude either upon direct RyR1 stimulation or in response to membrane depolarization. This study is the first demonstration of the potential of exon-skipping strategy for the therapy of central core disease, from the molecular to the functional level.


Sustained effects of once-a-week gait training with hybrid assistive limb for rehabilitation in chronic stroke: case study.
[Purpose] The purpose of this study was to investigate the accumulated and sustained effects of once-a-week gait training with a powered exoskeleton suit, Hybrid Assistive Limb, in a subject with chronic stroke. [Subject and Methods] The subject was a woman in her early sixties who had stroke onset approximately 5 years ago. A single-case ABA design was used. A 2-month baseline period was followed by an 8-week period of weekly gait training and a subsequent 2-month follow-up period. Throughout the study period, she underwent conventional physiotherapy. Outcome measures were the 10-meter walking test, timed up and go test, functional reach test, two-step test, and Berg Balance Scale. [Results] Significant improvements were seen in all outcome measures during the gait training period. Improvements in all outcome measures except walking speed were maintained at follow-up. [Conclusion] Continued gait training with Hybrid Assistive Limb once a week can improve gait and balance performance in patients with chronic stroke, and these improvements are maintained at least for two months.


Outbreak of a Leaf Disease Caused by Pseudocercospora ceratoniae on Carob in Sicily.
Carob (Ceratonia siliqua L.), an evergreen tree typical of the Mediterranean flora, has been grown in Sicily (Italy) from time immemorial for its fruits, used mainly as food for cattle and horses as well as for industrial production of alcohol. Although the economic importance of carob as a commercial crop has declined over the last decades, carob trees still are a characteristic aspect of the landscape in southeastern Sicily. In early April 1998, a severe outbreak of a foliar disease was noted on carob trees in the Ragusa province. Symptoms initially consisted of small (2 to 3 mm wide) dark brown, vein-limited spots, visible on both sides of the leaf and, later in the season, surrounded by a pale halo. In a humid atmosphere, spots scattered over the leaf blade but usually were most numerous along the midrib, enlarged, and coalesced, forming large blotches. Severely affected leaflets dropped, leaving the petiole attached to the tree. As a result, the trees appeared defoliated. Severely defoliated trees did not produce fruits. The causal agent of this disease was identified as Pseudocercospora ceratoniae (Pat. & Trab.) Deighton (1), an hyphomycetous fungus reported previously as a pathogen of carob under the name Cercospora ceratoniae Pat. & Trab. (3). On carob leaflets, P. ceratoniae formed grayish caespituli, confined to the lower surface of the necrotic spots. Caespituli consisted of dense fascicles of conidiophores (up to more than 50 conidiophores per fascicle) emerging through the stomata. Conidiophores were simple, slightly ampulliform, and geniculate at the conidial scars, which were conspicuous and unthickened. Old scars often were situated laterally on a short denticle. Conidia, borne singly as terminal blastospores and varying considerably in length, were pluriseptate, filiform, substraight or slightly curved, frequently slightly obclavate, with an obtuse apex and a short constriction at the base toward the truncate, unthickened hilum. Conidia from pure cultures of the fungus grown on water agar under black light were suspended in water and sprayed onto pot-grown carob plants. Inoculated plants were kept in a moist chamber for 48 h and subsequently transferred to the greenhouse. After 12 to 14 days leaf spots similar to those observed on naturally infected trees developed on inoculated plants and the pathogen was reisolated. Control plants sprayed with distilled water remained symptomless. C. ceratoniae had been recorded previously on carob in various Mediterranean countries, including Italy (2), but since has attracted little attention, being regarded as a sporadically occurring pathogen. Both mild temperatures during the winter and exceptionally frequent and persistent rain during the spring may have favored the epidemic outbreak of the disease caused by this fungus. References: (1) F. C. Deighton. 1976. Mycol. Pap. No. 140. Commonw. Mycol. Inst., Kew, England. (2) R. Parisi. Boll. Orto Bot. R. Univ. Napoli 10:155, 1932. (3) L. Roger. 1953. Phytopathologie des Pays Chauds. Vol. 2. P. Lechevalier, Paris.


Protein conjugate polysaccharide vaccines: challenges in development and global implementation.
Pneumonia and meningitis caused by Haemophilus influenzae type b, Streptococcus pneumoniae, and Neisseria meningitidis are among the leading causes of under five mortality and morbidity. Polysaccharide vaccines to prevent these infections are available since 1980s, but these are not effective in infants and children who are the common targets; therefore, protein conjugated were developed. The aim of this article is to understand the need for peumococcal protein conjugate vaccines, the challenges related to their development and global implementation, and the impact of these vaccines on global child health. Challenges in development of new vaccines are as follows:While pneumonia is a major threat in developing countries, available vaccine 7-valent pneumococcal conjugate vaccine (PCV7) protects against only 30% of invasive disease.Serogroup B of Neisseria meningitidis causes 32% of the cases in the USA and 45-80% or more in Europe. Due to similarity of its capsular polysaccharide with the cell surface glycoprotein on fetal brain tissue, developing a vaccine against this bacterium remains a challenge.Challenges in implementation are as follows:Replacement by nonvaccine serotypes;capsule switching;time duration of the antibody protective effect following vaccination;costs of the vaccines, programme costs, lack of knowledge of the disease burden, and targeting population groups for vaccination.


Distinct control sites located upstream from the levansucrase gene of Bacillus subtilis.
The sacR regulatory region, which modulates the expression of sacB, the structural gene for levansucrase, was separated into two parts: an upstream region which carries a constitutive promoter and a downstream region which carries a palindromic structure. Three types of fusions were constructed in which the aphA3 gene coding for kanamycin resistance of Streptococcus faecalis was placed downstream from different deleted sacR regions. Other fusions were constructed by inserting a promoter from phage SPO1 upstream from the sacB gene and part of the sacA region. A third kind of fusion was constructed in which the palindromic structure was flanked by a heterologous promoter and a heterologous structural gene. After introduction of these fusions into the chromosomal DNA of mutants affected in sacB regulation, it was possible to reveal different targets for the regulatory genes sacU, sacQ and sacS: the sacU and sacQ genes act on a region located near or just upstream from the promoter, and the sacS gene, which is involved in the induction process, acts on the palindromic structure.


Simplified immobilisation method for histidine-tagged enzymes in poly(methyl methacrylate) microfluidic devices.
Poly(methyl methacrylate) (PMMA) microfluidic devices have become promising platforms for a wide range of applications. Here we report a simple method for immobilising histidine-tagged enzymes suitable for PMMA microfluidic devices. The 1-step-immobilisation described is based on the affinity of the His-tag/Ni-NTA interaction and does not require prior amination of the PMMA surface, unlike many existing protocols. We compared it with a 3-step immobilisation protocol involving amination of PMMA and linking NTA via a glutaraldehyde cross-linker. These methods were applied to immobilise transketolase (TK) in PMMA microfluidic devices. Binding efficiency studies showed that about 15% of the supplied TK was bound using the 1-step method and about 26% of the enzyme was bound by the 3-step method. However, the TK-catalysed reaction producing l-erythrulose performed in microfluidic devices showed that specific activity of TK in the device utilising the 1-step immobilisation method was approximately 30% higher than that of its counterpart. Reusability of the microfluidic device produced via the 1-step method was tested for three cycles of enzymatic reaction and at least 85% of the initial productivity was maintained. The device could be operated for up to 40 h in a continuous flow and on average 70% of the initial productivity was maintained. The simplified immobilisation method required fewer chemicals and less time for preparation of the immobilised microfluidic device compared to the 3-step method while achieving higher specific enzyme activity. The method represents a promising approach for the development of immobilised enzymatic microfluidic devices and could potentially be applied to combine protein purification with immobilisation.


Duplex ultrasound screening detects high rates of deep vein thromboses in critically ill trauma patients.
American College of Chest Physician (ACCP) guidelines stratify deep venous thrombosis (DVT) risk in trauma patients based on injury pattern and pharmacologic prophylaxis. Screening is only recommended for patients with high-risk injuries who are unable to receive pharmacologic prophylaxis. However, the prevalence of lower extremity DVT (LEDVT) in trauma patients may be higher than reported in previous studies as many studies on DVT screening have not investigated calf vein DVTs (CVDVT) and have not exclusively targeted critically ill patients. Given that current ACCP guidelines recommend treatment of CVDVTs, we investigated the efficacy of duplex ultrasound (DUS) screening in critically ill trauma patients for all LEDVTs, including CVDVT, regardless of injury pattern, risk factors, or pharmacologic prophylaxis. Medical records of 264 intensive care unit trauma patients who received DUS screening for LEDVT were retrospectively examined for the presence of injuries conferring high risk for LEDVT, patient specific DVT risk factors, and low molecular weight heparin (LMWH) prophylaxis. Forty (15.2%) patients had LEDVTs found on DUS screening, 24 (60%) were CVDVT, and 30% of all DVTs were diagnosed within 1 week of admission. Patients without high-risk injuries receiving LMWH had a 13.5% DVT rate, which did not differ significantly from the 19.7% DVT rate in high-risk injury patients not receiving LMWH (P = .667). Lower extremity DVT is common in critically ill trauma patients, particularly in the first week following injury, regardless of injury pattern, DVT risk factors, or pharmacologic prophylaxis. Previous studies have underestimated DVT rates by not investigating CVDVTs and not exclusively targeting critically ill patients. We recommend early and continued DUS DVT screening of all critically ill trauma patients.


Sustained attention in a counting task: normal performance and functional neuroanatomy.
We examined changes in relative cerebral flood flow (relCBF) using PET during a sustained attention paradigm which included auditory stimulation and different tasks of mental counting. Ten normal volunteers underwent PET (15O water) during a baseline state and under experimental conditions which included listening to clicks, serial counting with auditory stimulation, counting with no auditory stimulation, and an additional component of working memory and time estimation. All subjects performed within normal limits in a battery of neurocognitive tests, which included measures of attention and working memory. Both counting with auditory stimulation and counting with no auditory stimulation engaged motor cortex, putamen, cerebellum, and anterior cingulate. Furthermore, counting with no auditory stimulation relative to counting while listening resulted in significantly increased relCBF in the inferior parietal, dorsolateral prefrontal, and anterior cingulate. The findings obtained in this study support the notion that the parietal and dorsolateral prefrontal cortex are involved when time estimation and working memory are taking part in a task requiring sustained attention.


Ion distribution measured by electron probe X-ray microanalysis in apoplastic and symplastic pathways in root cells in sunflower plants grown in saline medium.
Little is known about how salinity affects ions distribution in root apoplast and symplast. Using x-ray microanalysis, ions distribution and the relative contribution of apoplastic and symplastic pathways for delivery of ions to root xylem were studied in sunflower plants exposed to moderate salinity (EC=6). Cortical cells provided a considerably extended Na(+) and Cl(-) storage facility. Their contents are greater in cytoplasm (root symplast) as compared to those in intercellular spaces (root apoplast). Hence, in this level of salinity, salt damage in sunflower is not dehydration due to extracellular accumulation of sodium and chloride ions, as suggested in the Oertli hypothesis. On the other hand, reduction in calcium content due to salinity in intercellular space is less than reduction in the cytoplasm of cortical cells. It seems that sodium inhibits the radial movement of calcium in symplastic pathway more than in the apoplastic pathway. The cell wall seems to have an important role in providing calcium for the apoplastic pathway. Redistribution of calcium from the cell wall to intercellular space is because of its tendency towards xylem through the apoplastic pathway. This might be a strategy to enhance loading of calcium to xylem elements and to reduce calcium deficiency in young leaves under salinity. This phenomenon may be able to increase salt tolerance in sunflower plants. Supplemental calcium has been found to be effective in reducing radial transport of Na(+) across the root cells and their loading into the xylem, but not sodium absorption. Supplemental calcium enhanced Ca(2+) uptake and influx into roots and transport to stele.


The application of Aquaplast Thermoplastic as a bolus material in the radiotherapy of a patient with classic Kaposi's sarcoma at the lower extremity.
The classic Kaposi's sarcoma (KS) patients usually present with progressive skin lesions over the lower extremities, following an indolent course. Although radiotherapy is an effective treatment for KS, radiation over these lesions is not without difficulty. The intrinsic difficulty resides in how a homogenous radiation dose over superficial lesions involving large areas and irregular surface is properly delivered. Several bolus techniques have thus been invented. The Aquaplast RT Thermoplastic is a new type of bolus material that can be easily molded and conformed to the curvature of skin, with the equivalence to soft tissue in radiation interaction. This material was applied as the bolus for the irradiation of a classic KS patient, whose disease involved multiple skin areas over the right heel and ankle. Large parallel-opposed irradiation fields delivered by 60Co were used. Computed tomography demonstrated a close conformity of the bolus built-up by Aquaplast RT Thermoplastic to the surface of the ankle and foot. A dosimetry measurement further confirmed an adequate and homogenous distribution of desired dose around the lesions of the lower extremity. After a total dose of 39 Gy, divided in 13 fractions, the lesions remitted completely. Our data suggest that the use of Aquaplast RT Thermoplastic as a bolus material is helpful in delivering adequate dose to skin lesions of the lower extremities.


Patterns of functional improvement after revision knee arthroplasty.
Despite the increase in the number of total knee arthroplasty revisions, outcomes of such surgery and their correlates are poorly understood. The aim of this study was to characterize patterns of functional improvement after revision total knee arthroplasty over a two-year period and to investigate factors that affect such improvement patterns. Three hundred and eight patients in need of revision surgery were enrolled into the study, conducted at seventeen centers, and 221 (71.8%) were followed for two years. Short Form-36 (SF-36), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and Lower-Extremity Activity Scale (LEAS) scores were collected at baseline and every six months for two years postoperatively. A piecewise general linear mixed model, which models correlation between repeated measures and estimates separate slopes for different follow-up time periods, was employed to examine functional improvement patterns. Separate regression slopes were estimated for the zero to twelve-month and the twelve to twenty-four-month periods. The slopes for zero to twelve months showed significant improvement in all measures in the first year. The slopes for twelve to twenty-four months showed deterioration in the scores of the WOMAC pain subscale (slope = 0.67 +/- 0.21, p < 0.01) and function subscale (slope = 1.66 +/- 0.63, p < 0.05), whereas the slopes of the other measures had plateaued. A higher number of comorbidities was consistently the strongest deterrent of functional improvement across measures. The modes of failure of the primary total knee arthroplasty were instrument-specific predictors of outcome (for example, tibial bone lysis affected only the SF-36 physical component score [coefficient = -5.46 +/- 1.91, p < 0.01], while malalignment affected both the SF-36 physical component score [coefficient = 5.41 +/- 2.35, p < 0.05] and the LEAS score [coefficient = 1.42 +/- 0.69, p < 0.05]). Factors related to the surgical technique did not predict outcomes. The onset of worsening pain and knee-specific function in the second year following revision total knee arthroplasty indicates the need to closely monitor patients, irrespective of the mode of failure of the primary procedure or the surgical technique for the revision. This information may be especially important for patients with multiple comorbidities.


Changes in the Disparity Vergence Main Sequence after Treatment of Symptomatic Convergence Insufficiency in Children.
This study investigates the underlying physiological mechanisms that may lead to improved outcomes for symptomatic convergence insufficiency (Cl) patients after 12 weeks of office-based vergence/accommodation therapy (OBVAT) by evaluating the change in the main sequence of vergence and saccadic eye movements. In this prospective trial, 12 participants with symptomatic Cl were recruited and treated with 12 weeks of OBVAT. Outcome measures included the objective assessment of the following: peak velocity, time to peak velocity, latency, response amplitude, and clinical changes in the near point of convergence (NPC), positive fusional vergence (PFV) and symptoms via the Convergence Insufficiency Symptom Survey (CISS). Ten of the twelve participants (83%) were categorized as "successful" and two were "improved" based on pre-determined published criteria (CISS, NPC, PFV). There were statistically significant changes in peak velocity, time to peak velocity, and response amplitude for both 4° and 6° symmetrical convergence and divergence eye movements. There was a significant change in the main sequence ratio for convergence post-OBVAT compared to baseline measurements (P=0.007) but not for divergence or saccadic responses. Phasic/step vergence movements adjust the underlying neural control of convergence and are critical within a vision therapy program for Cl patients.


A stochastic model of latently infected cell reactivation and viral blip generation in treated HIV patients.
Motivated by viral persistence in HIV+ patients on long-term anti-retroviral treatment (ART), we present a stochastic model of HIV viral dynamics in the blood stream. We consider the hypothesis that the residual viremia in patients on ART can be explained principally by the activation of cells latently infected by HIV before the initiation of ART and that viral blips (clinically-observed short periods of detectable viral load) represent large deviations from the mean. We model the system as a continuous-time, multi-type branching process. Deriving equations for the probability generating function we use a novel numerical approach to extract the probability distributions for latent reservoir sizes and viral loads. We find that latent reservoir extinction-time distributions underscore the importance of considering reservoir dynamics beyond simply the half-life. We calculate blip amplitudes and frequencies by computing complete viral load probability distributions, and study the duration of viral blips via direct numerical simulation. We find that our model qualitatively reproduces short small-amplitude blips detected in clinical studies of treated HIV infection. Stochastic models of this type provide insight into treatment-outcome variability that cannot be found from deterministic models.


jmzTab-M: A Reference Parser, Writer, and Validator for the Proteomics Standards Initiative mzTab 2.0 Metabolomics Standard.
mzTab 2.0 for metabolomics (mzTab-M) is the most recent standard format developed in collaboration by the Proteomics and Metabolomics Standards Initiatives including contributions by the recently founded Lipidomics Standards Initiative. mzTab-M is a redesign of the original mzTab format which was geared toward reporting of proteomics results and, as such, provided only limited support for metabolites. As a tab-delimited, spreadsheet-like format, mzTab-M captures experimental metadata, summary information on small molecules across assays, MS features as a basis for quantitation, and evidence to support the reporting of individual or feature group identifications. Here, we present the Java reference implementation for reading, writing, and validating mzTab-M files. Furthermore, we provide a web application for conveniently validating mzTab-M files by a graphical user interface, and a command line validator that accompanies the library. The jmzTab-M library, version 1.0.4 ( https://doi.org/10.5281/zenodo.3362151 ), is available at https://github.com/lifs-tools/jmzTab-m and from Maven Central at https://search.maven.org/search?q=jmztabm under the terms of the open source Apache License 2.0. The web application as well as the Python and R implementations are available at https://github.com/lifs-tools . The respective Web sites link to additional API documentation, as well as to usage examples.


The B subunit of Escherichia coli heat-labile toxin alters the development and antigen-presenting capacity of dendritic cells.
Escherichia coli's heat-labile enterotoxin (Etx) and its non-toxic B subunit (EtxB) have been characterized as adjuvants capable of enhancing T cell responses to co-administered antigen. Here, we investigate the direct effect of intravenously administered EtxB on the size of the dendritic and myeloid cell populations in spleen. EtxB treatment appears to enhance the development and turnover of dendritic and myeloid cells from precursors within the spleen. EtxB treatment also gives a dendritic cell (DC) population with higher viability and lower activation status based on the reduced expression of MHC-II, CD80 and CD86. In this respect, the in vivo effect of EtxB differs from that of the highly inflammatory mediator lipopolysaccharide. In in vitro bone marrow cultures, EtxB treatment was also found to enhance the development of DC from precursors dependent on Flt3L. In terms of the in vivo effect of EtxB on CD4 and CD8 T cell responses in mice, the interaction of EtxB directly with DC was demonstrated following conditional depletion of CD11c(+) DC. In summary, all results are consistent with EtxB displaying adjuvant ability by enhancing the turnover of DC in spleen, leading to newly mature myeloid and DC in spleen, thereby increasing DC capacity to perform as antigen-presenting cells on encounter with T cells.


Partial genome sequence of Thioalkalivibrio thiocyanodenitrificans ARhD 1(T), a chemolithoautotrophic haloalkaliphilic sulfur-oxidizing bacterium capable of complete denitrification.
Thioalkalivibrio thiocyanodenitrificans strain ARhD 1(T) is a motile, Gram-negative bacterium isolated from soda lakes that belongs to the Gammaproteobacteria. It derives energy for growth and carbon fixation from the oxidation of sulfur compounds, most notably thiocyanate, and so is a chemolithoautotroph. It is capable of complete denitrification under anaerobic conditions. The draft genome sequence consists of 3,746,647 bp in 3 scaffolds, containing 3558 protein-coding and 121 RNA genes. T. thiocyanodenitrificans ARhD 1(T) was sequenced as part of the DOE Joint Genome Institute Community Science Program.


Time of Transfer of Admitted Patients from the ED: A Contributor to ED Boarding in High-Volume Community Hospitals.
The twin challenges of bed boarding and "hallway medicine" have emerged in recent years as key healthcare issues. Many hospitals, challenged with increasing demand and limited resources, have tried to find efficiencies within their operations. One such strategy is that of early morning discharges and expedited bed turnaround times. We conducted a retrospective study within three high-volume hospitals in the Greater Toronto Area looking at discharge times of in-patients and transfer times of admitted, Emergency Department (ED)-boarded patients. We discovered a consistent pattern of late-in-the-day discharges, and even later-in-the-day transfers of boarded ED patients, indicating that this may be a potential source of increased efficiency for overburdened hospitals.