Large transition state stabilization from a weak hydrogen bond.
A series of molecular rotors was designed to study and measure the rate accelerating effects of an intramolecular hydrogen bond. The rotors form a weak neutral O-H⋯O[double bond, length as m-dash]C hydrogen bond in the planar transition state (TS) of the bond rotation process. The rotational barrier of the hydrogen bonding rotors was dramatically lower (9.9 kcal mol-1) than control rotors which could not form hydrogen bonds. The magnitude of the stabilization was significantly larger than predicted based on the independently measured strength of a similar O-H⋯O[double bond, length as m-dash]C hydrogen bond (1.5 kcal mol-1). The origins of the large transition state stabilization were studied via experimental substituent effect and computational perturbation analyses. Energy decomposition analysis of the hydrogen bonding interaction revealed a significant reduction in the repulsive component of the hydrogen bonding interaction. The rigid framework of the molecular rotors positions and preorganizes the interacting groups in the transition state. This study demonstrates that with proper design a single hydrogen bond can lead to a TS stabilization that is greater than the intrinsic interaction energy, which has applications in catalyst design and in the study of enzyme mechanisms.


Gap junctions in developing thalamic and neocortical neuronal networks.
The presence of direct, cytoplasmatic, communication between neurons in the brain of vertebrates has been demonstrated a long time ago. These gap junctions have been characterized in many brain areas in terms of subunit composition, biophysical properties, neuronal connectivity patterns, and developmental regulation. Although interesting findings emerged, showing that different subunits are specifically regulated during development, or that excitatory and inhibitory neuronal networks exhibit various electrical connectivity patterns, gap junctions did not receive much further interest. Originally, it was believed that gap junctions represent simple passageways for electrical and biochemical coordination early in development. Today, we know that gap junction connectivity is tightly regulated, following independent developmental patterns for excitatory and inhibitory networks. Electrical connections are important for many specific functions of neurons, and are, for example, required for the development of neuronal stimulus tuning in the visual system. Here, we integrate the available data on neuronal connectivity and gap junction properties, as well as the most recent findings concerning the functional implications of electrical connections in the developing thalamus and neocortex.


Elimination of manganese(II,III) oxidation in Pseudomonas putida GB-1 by a double knockout of two putative multicopper oxidase genes.
Bacterial manganese(II) oxidation impacts the redox cycling of Mn, other elements, and compounds in the environment; therefore, it is important to understand the mechanisms of and enzymes responsible for Mn(II) oxidation. In several Mn(II)-oxidizing organisms, the identified Mn(II) oxidase belongs to either the multicopper oxidase (MCO) or the heme peroxidase family of proteins. However, the identity of the oxidase in Pseudomonas putida GB-1 has long remained unknown. To identify the P. putida GB-1 oxidase, we searched its genome and found several homologues of known or suspected Mn(II) oxidase-encoding genes (mnxG, mofA, moxA, and mopA). To narrow this list, we assumed that the Mn(II) oxidase gene would be conserved among Mn(II)-oxidizing pseudomonads but not in nonoxidizers and performed a genome comparison to 11 Pseudomonas species. We further assumed that the oxidase gene would be regulated by MnxR, a transcription factor required for Mn(II) oxidation. Two loci met all these criteria: PputGB1_2447, which encodes an MCO homologous to MnxG, and PputGB1_2665, which encodes an MCO with very low homology to MofA. In-frame deletions of each locus resulted in strains that retained some ability to oxidize Mn(II) or Mn(III); loss of oxidation was attained only upon deletion of both genes. These results suggest that PputGB1_2447 and PputGB1_2665 encode two MCOs that are independently capable of oxidizing both Mn(II) and Mn(III). The purpose of this redundancy is unclear; however, differences in oxidation phenotype for the single mutants suggest specialization in function for the two enzymes.


Optimal oxygenation of extremely low birth weight infants: a meta-analysis and systematic review of the oxygen saturation target studies.
The optimal oxygen saturation for extremely low birth weight infants in the postnatal period beyond the delivery room is not known. To summarize and discuss the results of the randomized trials, constituting the NEOPROM (Neonatal Oxygenation Prospective Meta-analysis) collaborative study, examining the effect of low versus high functional oxygen saturation targets in the postnatal period in premature infants with gestational age <28 weeks. A meta-analysis of SUPPORT (Surfactant, Positive Pressure and Pulse Oximetry Randomized Trial), the three BOOST II (Benefits of Oxygen Saturation Targeting) studies and the COT (Canadian Oxygen Trial) was performed including a total of 4,911 infants randomized to either a low (85-89%) or high (91-95%) functional oxygen saturation (SpO2) within the first 24 h after birth. Relative risks (RR; 95% CIs) comparing a low versus a high oxygen saturation target were 1.41 (1.14-1.74) for mortality at discharge or at follow-up, 0.74 (0.59-0.92) for severe retinopathy of prematurity, 0.95 (0.86-1.04) for physiologic bronchopulmonary dysplasia, 1.25 (1.05-1.49) for necrotizing enterocolitis, 1.02 (0.88-1.19) for brain injury, and 1.01 (0.95-1.08) for patent ductus arteriosus. RR >1.0 favors a high oxygen saturation. RRs for mortality and necrotizing enterocolitis are significantly increased and severe retinopathy of prematurity significantly reduced in low compared to high oxygen saturation target infants. There are no differences regarding physiologic bronchopulmonary dysplasia, brain injury or patent ductus arteriosus between the groups. Based on these results, it is suggested that functional SpO2 should be targeted at 90-95% in infants with gestational age <28 weeks until 36 weeks' postmenstrual age. However, there are still several unanswered questions in this field.


Methotrexate Efficacy in the Tight Control in Psoriatic Arthritis Study.
Methotrexate (MTX) is a commonly used disease-modifying antirheumatic drug in psoriatic arthritis, but there is conflicting evidence to support its efficacy. Within the Tight Control of Psoriatic Arthritis (TICOPA) study, patients were treated with MTX as part of the tight control protocol or standard care. Outcomes were recorded at the 12-week visit, including joint counts, skin, nail, enthesitis, dactylitis, and patient-reported measures. Of the 206 patients enrolled, 188 received MTX in the first 12 weeks of the trial with 104 receiving a mean dose > 15 mg/week. The proportions of patients achieving the American College of Rheumatology (ACR) outcomes at 12 weeks were ACR20 40.8%, ACR50 18.8%, and ACR70 8.6%, with 22.4% achieving minimal disease activity. Improvements were seen in psoriasis with 27.2% reaching a Psoriasis Area and Severity Index (PASI) 75. The proportion of patients with dactylitis and Leeds dactylitis instrument (LDI) scores decreased significantly (62.7% decrease in patients with dactylitis, median change LDI -59.7, -157.4 to -26.4, p = 0.033). The decrease in proportion of patients with enthesitis (25.7%) was significant, but the median change in enthesitis score was 0. There was a trend to higher proportions of patients receiving over 15 mg/week achieving ACR20, ACR50, and PASI75. Despite the open-label design of the data, improvements in multiple clinical outcomes were seen. The proportion of patients reaching ACR20 in the TICOPA study was higher than in the Methotrexate in Psoriatic Arthritis study (41% vs 34%), but no comparative data are available for other outcomes. There is a suggestion of a dose response, but this is hard to assess when patients doing well may be maintained on lower doses.


Scanning Tunneling Microscopy Observation of Phonon Condensate.
Using quantum tunneling of electrons into vibrating surface atoms, phonon oscillations can be observed on the atomic scale. Phonon interference patterns with unusually large signal amplitudes have been revealed by scanning tunneling microscopy in intercalated van der Waals heterostructures. Our results show that the effective radius of these phonon quasi-bound states, the real-space distribution of phonon standing wave amplitudes, the scattering phase shifts, and the nonlinear intermode coupling strongly depend on the presence of defect-induced scattering resonance. The observed coherence of these quasi-bound states most likely arises from phase- and frequency-synchronized dynamics of all phonon modes, and indicates the formation of many-body condensate of optical phonons around resonant defects. We found that increasing the strength of the scattering resonance causes the increase of the condensate droplet radius without affecting the condensate fraction inside it. The condensate can be observed at room temperature.


Adipose stromal vascular fraction attenuates TH1 cell-mediated pathology in a model of multiple sclerosis.
The therapeutic efficacy of adipose-derived stem cells (ASCs) has been investigated for numerous clinical indications, including autoimmune and neurodegenerative diseases. Less is known using the crude adipose product called stromal vascular fraction (SVF) as therapy, although our previous studies demonstrated greater efficacy at late-stage disease compared to ASCs in the experimental autoimmune encephalomyelitis (EAE) mouse, a model of multiple sclerosis. In this study, SVF cells and ASCs were administered during the pathogenic progression, designated as early disease, to elucidate immunomodulatory mechanisms when high immune cell activities associated with autoimmune signaling occur. These implications are essential for clinical translation when considering timing of administration for cell therapies. We investigated the effects of SVF cells and ASCs by analyzing the spleens, peripheral blood, and central nervous system tissues throughout the course of EAE disease following administration of SVF cells, ASCs, or vehicle. In vitro, immunomodulatory potentials of SVF cells and ASCs were measured when exposed to EAE-derived splenocytes. Interestingly, treatment with SVF cells and ASCs transiently enhanced the severity of disease directly after administration, substantiating this critical immunomodulatory signaling. More importantly, it was only the EAE mice treated with SVF cells that were able to overcome the advancing pathogenesis and showed improvements by the end of the study. The frequency of lesions in spinal cords following SVF treatment correlated with diminished activities of the T helper type 1 cells, known effector cells of this disease. Co-cultures with splenocytes isolated from EAE mice revealed transcripts of interleukin-10 and transforming growth factor-β, known promoters of regulatory T cells, that were greatly expressed in SVF cells compared to ASCs, and expression levels of signaling mediators related to effector T cells were insignificant in both SVF cells and ASCs. This is the first evidence, to date, to elucidate a mechanism of action of SVF treatment in an inflammatory, autoimmune disease. Our data supports key immunomodulatory signaling between cell therapies and T cells in this T cell-mediated disease. Together, treatment with SVF mediated immunomodulatory effects that diminished effector cell activities, promoted regulatory T cells, and reduced neuroinflammation.


Medical marijuana laws and adolescent marijuana use in the USA from 1991 to 2014: results from annual, repeated cross-sectional surveys.
Adolescent use of marijuana is associated with adverse later effects, so the identification of factors underlying adolescent use is of substantial public health importance. The relationship between US state laws that permit marijuana for medical purposes and adolescent marijuana use has been controversial. Such laws could convey a message about marijuana acceptability that increases its use soon after passage, even if implementation is delayed or the law narrowly restricts its use. We used 24 years of national data from the USA to examine the relationship between state medical marijuana laws and adolescent use of marijuana. Using a multistage, random-sampling design with replacement, the Monitoring the Future study conducts annual national surveys of 8th, 10th, and 12th-grade students (modal ages 13-14, 15-16, and 17-18 years, respectively), in around 400 schools per year. Students complete self-administered questionnaires that include questions on marijuana use. We analysed data from 1 098 270 adolescents surveyed between 1991 and 2014. The primary outcome of this analysis was any marijuana use in the previous 30 days. We used multilevel regression modelling with adolescents nested within states to examine two questions. The first was whether marijuana use was higher overall in states that ever passed a medical marijuana law up to 2014. The second was whether the risk of marijuana use changed after passage of medical marijuana laws. Control covariates included individual, school, and state-level characteristics. Marijuana use was more prevalent in states that passed a medical marijuana law any time up to 2014 than in other states (adjusted prevalence 15·87% vs 13·27%; adjusted odds ratio [OR] 1·27, 95% CI 1·07-1·51; p=0·0057). However, the risk of marijuana use in states before passing medical marijuana laws did not differ significantly from the risk after medical marijuana laws were passed (adjusted prevalence 16·25% vs 15·45%; adjusted OR 0·92, 95% CI 0·82-1·04; p=0·185). Results were generally robust across sensitivity analyses, including redefining marijuana use as any use in the previous year or frequency of use, and reanalysing medical marijuana laws for delayed effects or for variation in provisions for dispensaries. Our findings, consistent with previous evidence, suggest that passage of state medical marijuana laws does not increase adolescent use of marijuana. However, overall, adolescent use is higher in states that ever passed such a law than in other states. State-level risk factors other than medical marijuana laws could contribute to both marijuana use and the passage of medical marijuana laws, and such factors warrant investigation. US National Institute on Drug Abuse, Columbia University Mailman School of Public Health, New York State Psychiatric Institute.


The Role of MicroRNA-381 in Chondrogenesis and Interleukin-1-β Induced Chondrocyte Responses.
The molecular pathways regulating cartilage degradation are unclear. miR-381 was identified as a putative regulator of chondrogenesis related genes. Here, we examined its role in chondrogenesis and osteoarthritic cartilage degeneration. miR-381 expression was assessed in vitro in response to IL-1β stimulation in primary human (PHC) and mouse (PMC) chondrocytes, and ATDC5 derived chondrocytes; and in vivo in mouse embryos and human osteoarthritic cartilage. The effects of miR-381 on chondrogenesis and NF-kB signaling were assessed using a synthetic RNA mimic or inhibitor and luciferase assay, respectively. Upstream regulators of miR381 were probed using siRNA or overexpression plasmids for Sox9 and Runx2. miR-381 expression was elevated in chondrogenic and hypertrophic ATDC5 cells. miR-381 was induced in vitro by IL-1β in ATDC5 cells, PMCs, and PHCs, and was expressed in areas of cartilage degradation or absorption in vivo. Overexpression of Runx2 or Sox9 increased miR-381 expression in ATDC5 cells. miR-381 suppressed expression of collagen, type II, alpha 1, and enhanced expression of metalloproteinase-13 (MMP-13), but did not regulate NFKBIA and NKRF activity. miR-381 was highly expressed during chondrogenesis and in arthritic cartilage. It may contribute to absorption of the cartilage matrix by repressing type II collagen and inducing MMP-13.


Platelet glycoprotein IIb/IIIa inhibition with coronary stenting for acute myocardial infarction.
When administered in conjunction with primary coronary stenting for the treatment of acute myocardial infarction, a platelet glycoprotein IIb/IIIa inhibitor may provide additional clinical benefit, but data on this combination therapy are limited. We randomly assigned 300 patients with acute myocardial infarction in a double-blind fashion either to abciximab plus stenting (149 patients) or placebo plus stenting (151 patients) before they underwent coronary angiography. Clinical outcomes were evaluated 30 days and 6 months after the procedure. The angiographic patency of the infarct-related vessel and the left ventricular ejection fraction were evaluated at 24 hours and 6 months. At 30 days, the primary end point--a composite of death, reinfarction, or urgent revascularization of the target vessel--had occurred in 6.0 percent of the patients in the abciximab group, as compared with 14.6 percent of those in the placebo group (P=0.01); at 6 months, the corresponding figures were 7.4 percent and 15.9 percent (P=0.02). The better clinical outcomes in the abciximab group were related to the greater frequency of grade 3 coronary flow (according to the classification of the Thrombolysis in Myocardial Infarction trial) in this group than in the placebo group before the procedure (16.8 percent vs. 5.4 percent, P=0.01), immediately afterward (95.1 percent vs. 86.7 percent, P=0.04), and six months afterward (94.3 percent vs. 82.8 percent, P=0.04). One major bleeding event occurred in the abciximab group (0.7 percent); none occurred in the placebo group. As compared with placebo, early administration of abciximab in patients with acute myocardial infarction improves coronary patency before stenting, the success rate of the stenting procedure, the rate of coronary patency at six months, left ventricular function, and clinical outcomes.


Halocins and sulfolobicins: the emerging story of archaeal protein and peptide antibiotics.
Production of antibiotic peptides and proteins is a near-universal feature of living organisms regardless of phylogenetic classification. Bacteriocins (proteinaceous antimicrobials from the domain Bacteria) have been studied for over 75 years, and the eucaryocins (proteinaceous antimicrobials from the domain Eucarya) since the early 1960s. However, one domain of organisms, the Archaea, containing hyperthermophiles, extreme halophiles and the methanogens, is just beginning to be scrutinized for the production of peptide antibiotics. Production of archaeal proteinaceous antimicrobials (archaeocins) from extreme halophiles (halocins) is a nearly universal feature of the rod-shaped haloarchaea. Halocin activity is first detectable in culture supernatants at the beginning of the transition into stationary phase, concomitant with an induction of transcription of the structural gene. Halocins are diverse in size, consisting of proteins as large as 35 kDa and peptide "microhalocins" as small as 3.6 kDa. The 36 amino acids of microhalocin HalS8 are located in the interior of a 311-residue pro-protein from which they are liberated by an unknown mechanism. Microhalocins are hydrophobic and robust, withstanding heat, desalting and exposure to organic solvents. Unlike the peptide bacteriocins and the eucaryocins, microhalocins possess a large number of neutral residues and are not cationic, leaving their mechanism(s) of action mostly a mystery. While microhalocins affect a variety of haloarchaeal genera (kingdom Euryarchaeota), they also exhibit cross-kingdom toxicity, inhibiting or killing Sulfolobus species (kingdom Crenarchaeota). Finally, archaeocins also are produced by the hyperthermophile "Sulfolobus islandicus". These 20-kDa protein antibiotics are not excreted into the environment, but are associated with small particles apparently derived from the cell's S-layer.


Hydraulic driven fast and precise nonmagnetic tactile stimulator for neurophysiological and MEG measurements.
Electric stimulation of the peripheral nerves is well established as a diagnostic and research tool to analyze the somatosensory system. However, electric stimulation has some disadvantages. Electric stimulation of the median nerve triggers action potentials in all fiber populations of the nerve. Electric stimulation further creates artifacts and courses discomfort which is usually not well tolerated in the awake child. Therefore, the development of a more specific stimulation has constantly been a goal in recent years. There have been several approaches in the past to deliver somatic stimulation. However, all of them failed short in some aspects. In this study, a new type of somatosensory stimulator device was developed and compared against the gold standard of electric stimulation. The stimulation is achieved by repetitive tactile stimulation of the index finger using a blunt needle. In contrast to all previous approaches, we use a hydraulic system to move the needle up and downward. Given that water is very well suited to conduct pressure pulses it is possible to place the tactile stimulator device holding the needle close to the subject and the hydraulic driving system outside a critical area. Using a phantom, we showed that our stimulator is capable of delivering a stimulus precise on the submillisecond time scale. In addition, we test our stimulator on a healthy adult and compare the results against the electric stimulation. We can show the feasibility of measuring the electric responses of the peripheral nerve and while using MEG also the response of the primary somatosensory cortex. The tactile stimulation showed a more spatial focuses activation of the primary somatosensory cortex when compared against the electric stimulation. The proposed high-precision tactile stimulator will make it possible to analyze the somatosensory system noninvasively in children in the future.


The Arabidopsis ELF3 gene regulates vegetative photomorphogenesis and the photoperiodic induction of flowering.
Flowering in Arabidopsis thaliana is promoted by longday (LD) photoperiods such that plants grown in LD flower earlier, and after the production of fewer leaves, than plants grown in short-day (SD) photoperiods. The early-flowering 3 (elf3) mutant of Arabidopsis, which is insensitive to photoperiod with regard to floral initiation has been characterized elf3 mutants are also altered in several aspects of vegetative photomorphogenesis, including hypocotyl elongation. When inhibition of hypocotyl elongation was measured, elf3 mutant seedlings were less responsive than wild-type to all wavelengths of light, and most notably defective in blue and green light-mediated inhibition. When analyzed for the flowering-time phenotype, elf3 was epistatic to mutant alleles of the blue-light receptor encoding gene, HY4. However, when elf3 mutants were made deficient for functional phytochrome by the introduction of hy2 mutant alleles, the elf3 hy2 double mutants displayed the novel phenotype of flowering earlier than either single mutant while still exhibiting photoperiod insensitivity, indicating that a phytochrome-mediated pathway regulating floral initiation remains functional in elf3 single mutants. In addition, the inflorescences of one allelic combination of elf3 hy2 double mutants form a terminal flower similar to the structure produced by tfk1 single mutants. These results suggest that one of the signal transduction pathways controlling photoperiodism in Arabidopsis is regulated, at least in part, by photoreceptors other than phytochrome, and that the activity of the Arabidopsis inflorescence and floral meristem identity genes may be regulated by this same pathway.


Preventive dental care among Medicaid-enrolled senior adults: from community to nursing facility residence.
The primary objective of this study was to determine whether the utilization rate of preventive oral health care services while senior adults were community-dwelling differed from the rate after those same senior adults were admitted to nursing facilities. A secondary objective was to evaluate other significant predictors of receipt of preventive oral health procedures after nursing facility entry. Iowa Medicaid claims from 2007-2014 were accessed for adults who were 68+ years upon entry to a nursing facility and continuously enrolled in Medicaid for at least three years before and at least two years after admission (n = 874). Univariate, bivariate and multivariable analyses were conducted. During the five years that subjects were followed, 52.8% never received a dental exam and 75.9% never received a dental hygiene procedure. More Medicaid-enrolled senior adults received ≥1 preventive dental procedure in the two years while residing in a nursing facility compared to the three years before entry. In multivariable analyses, the strongest predictor of preventive oral health care utilization after entry was the receipt of preventive oral health services before entry (p < 0.01). The strongest predictor of receipt of dental procedures in the two years after nursing facility entry was the receipt of dental procedures in the three years before entry while community-dwelling. This underscores the importance of the senior adult establishing a source of dental care while community-dwelling.


Drought stress alters water relations and expression of PIP-type aquaporin genes in Nicotiana tabacum plants.
Plasma membrane intrinsic proteins (PIPs), a type of aquaporins, mediate water transport in many plant species. In this study, we investigated the relationship between the functions of PIP-type water channels and water relations of tobacco plants (Nicotiana tabacum cv. Samsun) under drought stress. Drought stress treatments have led to reductions in the stomatal conductance, transpiration, water potential and turgor pressure in leaves, and also the sap flow rate and osmotic hydraulic conductance in roots. In contrast, leaf osmotic pressure was increased in response to drought stress. Interestingly, the accumulation of NtPIP1;1 and NtPIP2;1 transcripts was significantly decreased, but only that of the NtAQP1 transcript was increased under drought stress. Functional analysis using Xenopus laevis oocytes revealed that NtPIP2;1 shows marked water transport activity, but the activities of NtAQP1 and NtPIP1;1 are weak or almost negligible, respectively, when expressed alone. However, co-expression of NtPIP1;1 with NtPIP2;1 significantly enhanced water transport activity compared with that of NtPIP1;1- or NtPIP2;1-expressing oocytes, suggesting that these two aquaporins may function as a water channel, forming a heterotetramer. Heteromerization of NtPIP1;1 and NtPIP2;1 was also suggested by co-expression analyses of NtPIP1;1-GFP (green fluorescent protein) and NtPIP2;1 in Xenopus oocytes. Re-watering treatments recovered water relation parameters and the accumulation of the three NtPIP transcripts to levels similar to control conditions. These results suggest that NtPIP1;1 and NtPIP2;1 play an important role in water transport in roots, and that expression of NtPIP1;1 and NtPIP2;1 is down-regulated in order to reduce osmotic hydraulic conductance in the roots of tobacco plants under drought stress.


Extra- and intra-cellular accumulation of platinum group elements by the marine microalga, Chlorella stigmatophora.
To better understand the marine biogeochemistry of the platinum group elements (PGE), Rh(III), Pd(II) and Pt(IV) were added in combination and at ppb concentrations to cultures of the marine microalga, Chlorella stigmatophora, maintained in sea water at 15 °C and under 60 μmol m(-2) s(-1) PAR. The accumulation of PGE was established in short-term (24-h) exposures, and under varying conditions of algal biomass and PGE concentration, and in a longer-term exposure (156-h) by ICP-MS analysis of sea water and nitric acid digests and EDTA washes of the alga. In short-term exposures, and under all conditions, the extent of accumulation by C. stigmatophora was in the order: Rh > Pd >> Pt; and Pd was internalised (or resistant to EDTA extraction) to a considerably greater extent than Rh and Pt. Accumulation isotherms were quasi-linear up to added PGE concentrations of 30 μg L(-1) and all metals displayed a significant reduction in accumulation on a weight-normalised basis with increasing density (biomass) of C. stigmatophora, an effect attributed to the production of exudates able to stabilise metals in sea water through complexation. In the longer-term exposure, kinetic constraints on the reactivities of Rh and, in particular, Pt, resulted in final degrees of accumulation and internalisation by C. stigmatophora that were greatest for Rh and similar between Pd and Pt. Among the PGE, therefore, Rh is predicted to participate in biological removal and transport processes in the marine environment to the greatest extent while decoupling in the biogeochemistries of Pd and Pt is predicted in shorter-term or more transient processes.


Posterior vertebral column subtraction osteotomy: a novel surgical approach for the treatment of multiple recurrences of tethered cord syndrome.
Recurrent tethered cord syndrome (TCS) has been reported to develop in 5-50% of patients following initial spinal cord detethering operations. Surgery for multiple recurrences of TCS can be difficult and is associated with significant complications. Using a cadaveric tethered spinal cord model, Grande and colleagues demonstrated that shortening of the vertebral column by performing a 15-25-mm thoracolumbar osteotomy significantly reduced spinal cord, lumbosacral nerve root, and terminal filum tension. Based on this cadaveric study, spinal column shortening by a thoracolumbar subtraction osteotomy may be a viable alternative treatment to traditional surgical detethering for multiple recurrences of TCS. In this article, the authors describe the use of posterior vertebral column subtraction osteotomy (PVCSO) for the treatment of 2 patients with multiple recurrences of TCS. Vertebral column resection osteotomy has been widely used in the surgical correction of fixed spinal deformity. The PVCSO is a novel surgical treatment for multiple recurrences of TCS. In such cases, PVCSO may allow surgeons to avoid neural injury by obviating the need for dissection through previously operated sites and may reduce complications related to CSF leakage. The novel use of PVCSO for recurrent TCS is discussed in this report, including surgical considerations and techniques in performing PVCSO.


A simple model of transmitter release and facilitation.
We describe a model of synaptic transmitter release and presynaptic facilitation that is based on activation of release sites by single Ca2+ microdomains. Facilitation is due to Ca2+ that remains bound to release sites between impulses. This model is inherently stochastic, but deterministic equations can be derived for the mean release. The number of equations required to describe the mean release is small, so it is practical to use the model with models of neuronal electrical activity to investigate the effects of different input spike patterns on presynaptic facilitation. We use it in conjunction with a model of dopamine-secreting neurons of the basal ganglia to demonstrate that transmitter release is greater when the neuron bursts than when it spikes continuously, due to the greater facilitation generated by the bursting impulse pattern. Finally, a minimal form of the model is described that is coupled to simple models of postsynaptic receptors and passive membrane to compute the postsynaptic voltage response to a train of presynaptic stimuli. This form of the model is appropriate for neural network simulations.


Body mass index and socioeconomic position are associated with 9-year trajectories of multimorbidity: A population-based study.
Multimorbidity is a growing public health problem and is more common in women than men. However, little is known about multimorbidity trajectories, in terms of the accumulation of disease over time, or about the determinants of these trajectories. We sought to identify lifestyle and socioeconomic factors related to multimorbidity trajectories in mid-aged women. Participants were from the Australian Longitudinal Study on Women's Health, a nationally representative population-based study. We included 4865 women born 1946-51, without chronic disease in 1998, followed triennially for 12 years. We used latent class growth analysis to identify 9-year multimorbidity trajectories and multinomial regression to calculate relative risk ratios (RRRs) for associations between baseline lifestyle and socioeconomic factors and trajectories. We identified five multimorbidity trajectories: 'no morbidity, constant'; 'low morbidity, constant'; 'moderate morbidity, constant'; 'no morbidity, increasing'; and 'low morbidity, increasing'. Overweight and obesity were associated with an increased risk of the 'no morbidity, increasing' (RRR 1.70, 95% CI 1.16 to 2.50 and 2.69, 95% CI 1.69 to 4.28, respectively) and the 'low morbidity, increasing' (RRR 2.57, 95% CI 1.56 to 4.24 and 4.28, 95% CI 2.41 to 7.60, respectively) trajectories, as compared to the 'no morbidity, constant' group. Low education and difficulty managing on income were also associated with trajectories of poorer health. Among mid-aged women, overweight/obesity and lower socioeconomic status are major risk factors for trajectories characterised by accumulation of chronic disease. These highlight key target areas for preventive approaches aimed at reducing the risk of accumulation of morbidities in mid-aged women.


Calcium-dependent titin-thin filament interactions in muscle: observations and theory.
Gaps in our understanding of muscle mechanics demonstrate that the current model is incomplete. Increasingly, it appears that a role for titin in active muscle contraction might help to fill these gaps. While such a role for titin is increasingly accepted, the underlying molecular mechanisms remain unclear. The goals of this paper are to review recent studies demonstrating Ca2+-dependent interactions between N2A titin and actin in vitro, to explore theoretical predictions of muscle behavior based on this interaction, and to review experimental data related to the predictions. In a recent study, we demonstrated that Ca2+ increases the association constant between N2A titin and F-actin; that Ca2+ increases rupture forces between N2A titin and F-actin; and that Ca2+ and N2A titin reduce sliding velocity of F-actin and reconstituted thin filaments in motility assays. Preliminary data support a role for Ig83, but other Ig domains in the N2A region may also be involved. Two mechanical consequences are inescapable if N2A titin binds to thin filaments in active muscle sarcomeres: (1) the length of titin's freely extensible I-band should decrease upon muscle activation; and (2) binding between N2A titin and thin filaments should increase titin stiffness in active muscle. Experimental observations demonstrate that these properties characterize wild type muscles, but not muscles from mdm mice with a small deletion in N2A titin, including part of Ig83. Given the new in vitro evidence for Ca2+-dependent binding between N2A titin and actin, it is time for skepticism to give way to further investigation.


Stop flow lithography in perfluoropolyether (PFPE) microfluidic channels.
Stop Flow Lithography (SFL) is a microfluidic-based particle synthesis method for creating anisotropic multifunctional particles with applications that range from MEMS to biomedical engineering. Polydimethylsiloxane (PDMS) has been typically used to construct SFL devices as the material enables rapid prototyping of channels with complex geometries, optical transparency, and oxygen permeability. However, PDMS is not compatible with most organic solvents which limit the current range of materials that can be synthesized with SFL. Here, we demonstrate that a fluorinated elastomer, called perfluoropolyether (PFPE), can be an alternative oxygen permeable elastomer for SFL microfluidic flow channels. We fabricate PFPE microfluidic devices with soft lithography and synthesize anisotropic multifunctional particles in the devices via the SFL process--this is the first demonstration of SFL with oxygen lubrication layers in a non-PDMS channel. We benchmark the SFL performance of the PFPE devices by comparing them to PDMS devices. We synthesized particles in both PFPE and PDMS devices under the same SFL conditions and found the difference of particle dimensions was less than a micron. PFPE devices can greatly expand the range of precursor materials that can be processed in SFL because the fluorinated devices are chemically resistant to most organic solvents, an inaccessible class of reagents in PDMS-based devices due to swelling.


Transcranial direct current stimulation and working memory: Comparison of effect on learning shapes and English letters.
We present the results of a study investigating whether there is an effect of Anodal-Transcranial Direct Current Stimulation (A-tDCS) on working memory (WM) performance. The relative effectiveness of A-tDCS on WM is investigated using a 2-back test protocol using two commonly used memory visual stimuli (shapes and letters). In a double-blinded, randomised, crossover, sham-controlled experiment, real A-tDCS and sham A-tDCS were applied separately to the left dorsolateral prefrontal cortex (L-DLPFC) of twenty healthy subjects. There was a minimal interval of one week between sham and real A-tDCS sessions. For the letters based stimulus experiment, 2-back test recall accuracy was measured for a set of English letters (A-L) which were presented individually in a randomised order where each was separated by a blank interval. A similar 2-back protocol was used for the shapes based stimuli experiment where instead of letters, a set of 12 geometric shapes were used. The working memory accuracy scores measured appeared to be significantly affected by memory stimulus type used and by the application of A-tDCS (repeated measures ANOVA p<0.05). A large effect size (d = 0.98) and statistical significance between sham and real A-tDCS WM scores (p = 0.01) was found when shapes were used as a visual testing stimulus, while low (d = 0.38) effect size and insignificant difference (p = 0.15) was found when letters were used. This results are important as they show that recollection different stimuli used in working memory can be affected differently by A-tDCS application. This highlights the importance of considering using multiple methods of WM testing when assessing the effectiveness of A-tDCS.


In vitro hematopoiesis produces a distinct class of immature dendritic cells from spleen progenitors with limited T cell stimulation capacity.
The study of dendritic cells (DC) has been hampered by the difficulty of isolating rare cells for analysis of their phenotype and function. Interpretation of the DC lineage has been largely influenced by studies on cell populations which can be readily isolated and amplified in the presence of cytokines. Long term cultures (LTC) from murine spleen have been shown to support continuous in vitro hematopoiesis of DC dependent on interaction with a stromal cell monolayer. LTC-DC represent a single, stable class of DC derived by constant turnover of spleen DC progenitors maintained within stroma. They represent a resident DC population in spleen. The functional characteristics of LTC-DC have been studied in terms of capacity to stimulate T cells and response to activation by environmental stimuli. LTC-DC have many morphological, phenotypic and functional properties reflecting an immature or partially mature, marginal zone-like CD4(-)CD8(-) splenic DC subset. They are highly endocytic and can process and present protein antigen to naive hen egg lysozyme (HEL)-specific MHC-II-restricted TCR-Tg CD4(+) T cells. They do not, however, induce T cell proliferation in a mixed lymphocyte reaction. LTC-DC do not respond in a typical fashion to common DC activators like LPS and CD40L. They upregulate MHC-I and CD80/CD86 but not MHC-II and CD40. They reflect an endogenous, immature DC subset in spleen with properties distinct from immature DC located in peripheral tissues.


Multigenerational evaluation of sodium fluoride in rats.
Since the mid 1940s, fluoride has been added to tap water in American communities in an effort to reduce the incidence of dental caries in the population. When the levels of fluoride in drinking water were tested and set, water was the only measurable source of fluoride for most communities. Now, adults and children ingest fluoride with foods and beverages prepared with fluoridated water, and they are exposed to fluoride-containing dental products. As a result, exposure to fluoride is greater than had been anticipated. In the early 1990s, the existing reproductive studies were reviewed in several reports and were considered to be inadequate to determine potential reproductive or developmental hazards. The effects of sodium fluoride ingestion at 0, 25, 100, 175 or 250 ppm in drinking water measured in rats throughout three generations are reported here. Feed and fluid consumption, body weights and clinical signs were recorded at regular intervals. Decreased fluid consumption observed at 175 and 250 ppm was attributed to decreased palatability and did not affect reproduction. No cumulative effects were observed in the three generations. Mating, fertility and survival indices were not affected. Organ-to-body-weight ratios and organ-to-brain weight ratios were not affected. Sodium fluoride up to 250 ppm did not affect reproduction in rats.


Fenamates stimulate BKCa channel osteoblast-like MG-63 cells activity in the human.
The fenamates, a family of nonsteroidal anti-inflammatory drugs that are derivatives of N-phenylanthranilic acid, are the inhibitors of cyclo-oxygenase. The ionic mechanism of actions of these compounds in osteoblasts is not well understood. The effects of the fenamates on ionic currents were investigated in a human osteoblast-like cell line (MG-63) with the aid of the whole-cell and inside-out configurations of the patch-clamp technique. In MG-63 cells, niflumic acid and meclofenamic acid increased K+ outward currents (IK). The niflumic acid-stimulated IK was reversed by subsequent application of iberiotoxin or paxilline, yet not by that of glibenclamide or apamin. In the inside-out configuration, niflumic acid (30 micromol/L) added to the bath did not modify single-channel conductance but increased the activity of large-conductance Ca2+-activated K+ (BKCa) channels. The EC50 values for niflumic acid- and meclofenamic acid-induced channel activity were 22 and 24 micromol/L, respectively. Niflumic acid (30 micromol/L) and meclofenamic acid (30 micromol/L) shifted the activation curve of BKCa channels to less positive membrane potentials. Membrane stretch potentiated niflumic acid-stimulated channel activity. The rank order of potency for the activation of BKCa channels in these cells was niflumic acid = meclofenamic acid > tolfenamic acid > flufenamic acid > nimesulide. Evans blue and nordihydroguaiaretic acid increased channel activity; however, indomethacin, piroxicam, and NS-398 had no effect on it. The fenamates can stimulate BKCa channel activity in a manner that seems to be independent of the action of these drugs on the prostaglandin pathway. The activation of the BKCa channel may hyperpolarize the osteoblast, thereby modulating osteoblastic function.


Monte Carlo calculations of absorbed doses in tumours using a modified MOBY mouse phantom for pre-clinical dosimetry studies.
Clinical treatment with radionuclides is usually preceded by biokinetic and dosimetry studies in small animals. Evaluation of the therapeutic efficacy is essential and must rely on accurate dosimetry, which in turn must be based on a realistic geometrical model that properly describes the transport of radiation. It is also important to include the source distribution in the dosimetry calculations. Tumours are often implanted subcutaneously in animals, constituting an important additional source of radiation that often is not considered in the dosimetry models. The aims of this study were to calculate S values of the mouse, and determine the absorbed dose contribution to and from subcutaneous tumours inoculated at four different locations. The Moby computer program generates a three dimensional (3D) voxel-based phantom. Tumours were modelled as half-spheres on the body surface, and the radius was varied to study different tumour masses. The phantoms were used as input for Monte Carlo simulations of absorbed fractions and S factors with the radiation transport code MCNPX 2.6f. Calculations were performed for monoenergetic photons and electrons, and the radionuclides (125)I, (131)I, (111)In, (177)Lu and (90)Y. Electron energy and tumour size are important for both self- and cross-doses. If the activity is non-uniformly distributed within the body, the position of the tumour must be considered in order to calculate the tumour absorbed dose accurately. If the uptake in the tumour is high compared with that in adjacent organs the absorbed dose contribution to organs from the tumour cannot be neglected. In order to perform accurate tumour dosimetry in mouse models it is necessary to take the additional contribution from the activity distribution within the body of the mouse into account. This may be of significance in the interpretation of radiobiological tumour response in pre-clinical studies.


Self-reported willingness to share political news articles in online surveys correlates with actual sharing on Twitter.
There is an increasing imperative for psychologists and other behavioral scientists to understand how people behave on social media. However, it is often very difficult to execute experimental research on actual social media platforms, or to link survey responses to online behavior in order to perform correlational analyses. Thus, there is a natural desire to use self-reported behavioral intentions in standard survey studies to gain insight into online behavior. But are such hypothetical responses hopelessly disconnected from actual sharing decisions? Or are online survey samples via sources such as Amazon Mechanical Turk (MTurk) so different from the average social media user that the survey responses of one group give little insight into the on-platform behavior of the other? Here we investigate these issues by examining 67 pieces of political news content. We evaluate whether there is a meaningful relationship between (i) the level of sharing (tweets and retweets) of a given piece of content on Twitter, and (ii) the extent to which individuals (total N = 993) in online surveys on MTurk reported being willing to share that same piece of content. We found that the same news headlines that were more likely to be hypothetically shared on MTurk were also shared more frequently by Twitter users, r = .44. For example, across the observed range of MTurk sharing fractions, a 20 percentage point increase in the fraction of MTurk participants who reported being willing to share a news headline on social media was associated with 10x as many actual shares on Twitter. We also found that the correlation between sharing and various features of the headline was similar using both MTurk and Twitter data. These findings suggest that self-reported sharing intentions collected in online surveys are likely to provide some meaningful insight into what content would actually be shared on social media.


The GH3 family in plants: genome wide analysis in rice and evolutionary history based on EST analysis.
The GH3 gene family in Arabidopsis, implicated in hormonal homeostasis through the conjugation of indolacetic and jasmonic acids to amino acids, is involved in a broad range of plant growth and development processes. In this work, the analysis of the GH3 family in the genome of Oryza sativa identified 13 hypothetical ORFs. EST analysis and RT-PCR assays demonstrated that 12 of them were active genes. An extensive EST analysis of the GH3 family performed on 26 plant species was used to estimate the minimum number of GH3 genes en each one. The data indicated that the members of the GH3 family progressively increased in the different plant divisions from Chlorophyta (0), Bryophyta (3), and Coniferophyta (4), to Magnoliophyta (7-19). Phylogenetic analyses showed a high degree of conservation between Arabidopsis and rice GH3 proteins and, in general, in the plant kingdom. The data revealed a homology clustering consistent with the functional classification of the Arabidopsis proteins, since most of the 110 sequences analyzed grouped into 2 main clusters, corresponding to the Arabidopsis functional groups I (jasmonic acid adenylation) and II (indolacetic acid adenylation). And additional cluster including group III (non-adenylation ability) was exclusively composed of proteins from Arabidopsis thaliana, Brassica napus and Gossypium hirsutum.


Trends in quality of care and dying perceived by family caregivers of nursing home residents with dementia 2005-2019.
Dementia palliative care is increasingly subject of research and practice improvement initiatives. To assess any changes over time in the evaluation of quality of care and quality of dying with dementia by family caregivers. Combined analysis of eight studies with bereaved family caregivers' evaluations 2005-2019. Family caregivers of nursing home residents with dementia in the Netherlands (n = 1189) completed the End-of-Life in Dementia Satisfaction With Care (EOLD-SWC; quality of care) and Comfort Assessment in Dying (EOLD-CAD, four subscales; quality of dying) instruments. Changes in scores over time were analysed using mixed models with random effects for season and facility and adjustment for demographics, prospective design and urbanised region. The mean total EOLD-SWC score was 33.40 (SD 5.08) and increased by 0.148 points per year (95% CI, 0.052-0.244; adjusted 0.170 points 95% CI, 0.055-0.258). The mean total EOLD-CAD score was 30.80 (SD 5.76) and, unadjusted, there was a trend of decreasing quality of dying over time of -0.175 points (95% CI, -0.291 to -0.058) per year increment. With adjustment, the trend was not significant (-0.070 EOLD-CAD total score points, 95% CI, -0.205 to 0.065) and only the EOLD-CAD subscale 'Well being' decreased. We identified divergent trends over 14 years of increased quality of care, while quality of dying did not increase and well-being in dying decreased. Further research is needed on what well-being in dying means to family. Quality improvement requires continued efforts to treat symptoms in dying with dementia.


Effects of Short-Term Exercise Training With and Without Milk Intake on Cardiometabolic and Inflammatory Adaptations in Obese Adolescents.
There is some evidence that a combination of factors can reduce inflammation and associated metabolic risk factors. We studied the early cardiometabolic and inflammatory adaptations to a short-term exercise intervention with and without milk in obese adolescents. Fifty-four adolescents were randomized to consume milk post exercise (MILK) or a carbohydrate beverage (CONT) during one-week of daily exercise. Insulin levels were not different between the groups post training. Glucose was reduced over time in both groups (-9 ± 13 mg/ dl MILK and -6 ± 14 mg/dl CONT, p < .05) but not different between groups. There was a greater decrease in mean arterial pressure (MAP) in the MILK group (-3 ± 6 mmHg MILK vs. 2 ± 7 mmHg CONT, p < .04). Milk provided postexercise did not affect C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) or interleukin-6 (IL-6). The exercise intervention led to an increase in TNF-α in both groups (0.27 ± 0.7 pg/ml MILK and 0.48 ± 0.6 pg/ml CONT, p < .001). The early adaptations to a short-term exercise intervention in obese adolescents include a reduction in MAP and an increase in some inflammatory markers.


Reducing the expression of implicit stereotypes: reflexive control through implementation intentions.
The authors tested the effectiveness of implementation intentions as a strategy for limiting the behavioral expression of implicit stereotypes. Implementation intentions are if-then plans that link an intended response to an anticipated situational cue, thereby enabling a reflexive form of control. The authors examined whether two different types of implementation intentions could improve response accuracy on the Shooter Task, a reaction time measure of implicit stereotyping. In Study 1, participants used a distraction-inhibiting implementation intention designed to engage control over the perception of goal-irrelevant stimuli (e.g., race). In Study 2, participants used a response-facilitating implementation intention designed to promote goal-directed action. Across studies, implementation intentions improved accuracy, thereby limiting the behavioral expression of implicit stereotypes. Furthermore, process dissociation analyses indicated that the distraction-inhibiting implementation intention increased controlled processing while reducing automatic stereotype activation, whereas the response-facilitating implementation intention increased only controlled processing. Implications for goal strategy approaches to reducing prejudice are discussed.


Role of autophagy in IL-1β export and release from cells.
The autophagy pathway known also as macroautophagy (herein referred to as autophagy) is characterized by the formation of double-membrane organelles that capture cytosolic material. Based on pathway termination alternatives, autophagy has been divided into degradative and secretory. During degradative autophagy, autophagosomes typically fuse with lysosomes upon which the sequestered material is degraded. During secretory autophagy, instead of degradation the sequestered cargo is subjected to active secretion or passive release. In this review, we focus on the mechanisms of secretion/passive release of the potent pro-inflammatory cytokine IL-1β, as a prototypical leaderless cytosolic protein cargo studied in the context of secretory autophagy.


Effect of cations on the hydrated proton.
We report on a strong nonadditive effect of protons and other cations on the structural dynamics of liquid water, which is revealed using dielectric relaxation spectroscopy in the frequency range of 1-50 GHz. For pure acid solutions, protons are known to have a strong structuring effect on water, leading to a pronounced decrease of the dielectric response. We observe that this structuring is reduced when protons are cosolvated with salts. This reduction is exclusively observed for combinations of protons with other ions; for all studied solutions of cosolvated salts, the effect on the structural dynamics of water is observed to be purely additive, even up to high concentrations. We derive an empirical model that quantitatively describes the nonadditive effect of cosolvated protons and cations. We argue that the effect can be explained from the special character of the proton in water and that Coulomb fields exerted by other cations, in particular doubly charged cations like Mg(2+)aq and Ca(2+)aq, induce a localization of the H(+)aq hydration structures.


Development of interventions for the secondary prevention of Alzheimer's dementia: the European Prevention of Alzheimer's Dementia (EPAD) project.
Alzheimer's dementia affects more than 40 million people worldwide with substantial increases in prevalence anticipated. Interventions that either modify risk or reduce the development of early disease could delay the onset of dementia or reduce the rate of cognitive and functional decline. The European Prevention of Alzheimer's Dementia (EPAD) is a public-private consortium, funded by the Innovative Medicines Initiative, designed to increase the likelihood of successful development of new treatments for the secondary prevention of Alzheimer's dementia. EPAD will help with testing of different agents in this pre-dementia population through four components: improvement of access to existing cohorts and registries, development of the EPAD Registry of approximately 24,000 people who might be at increased risk of developing Alzheimer's dementia, establishment of the EPAD Longitudinal Cohort Study of 6000 people at any one time, and establishment of an adaptive, proof-of-concept trial including 1500 participants at any given time. The need for EPAD and its key design elements are described, and we discuss EPAD in relation to similar projects in progress. These parallel efforts reflect the need for a coordinated, worldwide battle against dementia, in which EPAD will play a crucial role.


A retrospective study of HIV pre-exposure prophylaxis counselling among non-Hispanic Black youth diagnosed with bacterial sexually transmitted infections in the United States, 2014-2019.
Youth account for a disproportionate number of new HIV infections; however, pre-exposure prophylaxis (PrEP) use is limited. We evaluated PrEP counselling rates among non-Hispanic Black youth in the United States after a bacterial sexually transmitted infection (STI) diagnosis. We conducted a retrospective cohort study of Black youth receiving care at two academically affiliated clinics in Philadelphia between June 2014 and June 2019. We compared PrEP counselling for youth who received primary care services versus those who did not receive primary care services, all of whom met PrEP eligibility criteria due to STI diagnosis per U.S. Centers for Disease Control and Prevention clinical practice guidelines. Two logistic regression models for receipt of PrEP counselling were fit: Model 1 focused on sexual and gender minority (SGM) status and Model 2 on rectal STIs with both models adjusted for patient- and healthcare-level factors. Four hundred and sixteen patients met PrEP eligibility criteria due to STI based on sex assigned at birth and sexual partners. Thirty patients (7%) had documentation of PrEP counselling. Receipt of primary care services was not significantly associated with receipt of PrEP counselling in either Model 1 (adjusted OR (aOR) 0.10 [95% CI 0.01, 0.99]) or Model 2 (aOR 0.52 [95% CI 0.10, 2.77]). Receipt of PrEP counselling was significantly associated with later calendar years of STI diagnosis (aOR 6.80 [95% CI 1.64, 29.3]), assigned male sex at birth (aOR 26.2 [95% CI 3.46, 198]) and SGM identity (aOR 317 [95% CI 39.9, 2521]) in Model 1 and later calendar years of diagnosis (aOR 3.46 [95% CI 1.25, 9.58]), assigned male sex at birth (aOR 18.6 [95% CI 3.88, 89.3]) and rectal STI diagnosis (aOR 28.0 [95% CI 8.07, 97.5]) in Model 2. Fourteen patients (3%) started PrEP during the observation period; 12/14 (86%) were SGM primary care patients assigned male sex at birth. PrEP counselling and uptake among U.S. non-Hispanic Black youth remain disproportionately low despite recent STI diagnosis. These findings support the need for robust investment in PrEP-inclusive sexual health services that are widely implemented and culturally tailored to Black youth, particularly cisgender heterosexual females.


Forced mobilization accelerates pathogenesis: characterization of a preclinical surgical model of osteoarthritis.
Preclinical osteoarthritis (OA) models are often employed in studies investigating disease-modifying OA drugs (DMOADs). In this study we present a comprehensive, longitudinal evaluation of OA pathogenesis in a rat model of OA, including histologic and biochemical analyses of articular cartilage degradation and assessment of subchondral bone sclerosis. Male Sprague-Dawley rats underwent joint destabilization surgery by anterior cruciate ligament transection and partial medial meniscectomy. The contralateral joint was evaluated as a secondary treatment, and sham surgery was performed in a separate group of animals (controls). Furthermore, the effects of walking on a rotating cylinder (to force mobilization of the joint) on OA pathogenesis were assessed. Destabilization-induced OA was investigated at several time points up to 20 weeks after surgery using Osteoarthritis Research Society International histopathology scores, in vivo micro-computed tomography (CT) volumetric bone mineral density analysis, and biochemical analysis of type II collagen breakdown using the CTX II biomarker. Expression of hypertrophic chondrocyte markers was also assessed in articular cartilage. Cartilage degradation, subchondral changes, and subchondral bone loss were observed as early as 2 weeks after surgery, with considerable correlation to that seen in human OA. We found excellent correlation between histologic changes and micro-CT analysis of underlying bone, which reflected properties of human OA, and identified additional molecular changes that enhance our understanding of OA pathogenesis. Interestingly, forced mobilization exercise accelerated OA progression. Minor OA activity was also observed in the contralateral joint, including proteoglycan loss. Finally, we observed increased chondrocyte hypertrophy during pathogenesis. We conclude that forced mobilization accelerates OA damage in the destabilized joint. This surgical model of OA with forced mobilization is suitable for longitudinal preclinical studies, and it is well adapted for investigation of both early and late stages of OA. The time course of OA progression can be modulated through the use of forced mobilization.


Sugar-sweetened beverage intake and serum testosterone levels in adult males 20-39 years old in the United States.
This population-based study was designed to investigate whether consumption of sugar-sweetened beverages (SSB) is associated with lower serum total testosterone concentration in men 20-39 years old. All data for this study were retrieved from the National Health and Nutrition Examination Survey (NHANES) 2011-2012. The primary outcome was serum testosterone concentration, and main independent variable was SSB intake. Other variables included age, race/ethnicity, poverty/income ratio, body mass index (BMI), serum cotinine, heavy drinking, and physical activity. Among all subjects (N = 545), 486 (90.4%) had normal testosterone levels (defined as ≥231 ng/dL) and 59 (9.6%) had low testosterone levels (defined as < 231 ng/dL). Multivariate logistic regression revealed the odds of low testosterone was significantly greater with increasing SSB consumption (Q4 [≥442 kcal/day] vs. Q1 [≤137 kcal/day]), adjusted odds ratio [aOR] = 2.29, p = 0.041]. After adjusting for possible confounding variables, BMI was an independent risk factor for low testosterone level; subjects with BMI ≥ 25 kg/m2 had a higher risk of having a low testosterone level than those with BMI < 25 kg/m2 (aOR = 3.68, p = 0.044). SSB consumption is significantly associated with low serum testosterone in men 20-39 years old in the United States.


Upconversion fluorescent and X-ray-sensitive bifunctional nanoprobes for assessing the penetrability of inorganic nanoparticles in the digestive system.
Nanotechnology is receiving increasing attention due to its fantastic advantages and potential applications in nanofood and nanomedicine. However, the safety of touching manufactured nanoparticles is still uncertain for human beings. Here, we track inorganic nanoparticles in the digestive system of the mouse through upconversion fluorescence and X-ray imaging, and try to demonstrate whether or not the inorganic nanoparticles will penetrate the digestive system to enter the blood system. Lanthanide-doped upconversion nanoparticles, which can convert infrared light to visible light and are simultaneously sensitive to X-rays, were selected as model inorganic nanoparticles. The investigation clarifies that even the ultrathin nanoparticles (∼5 nm) could not penetrate the digestive tract to enter the bloodstream or surrounding tissues, but were gradually excreted out. Our results help assess the safety of inorganic nanoparticles potentially used in nanofood and nanomedicine.


Repeatability and Diagnostic Value of Nasal Potential Difference in a Genetically Admixed Population.
The genetic diversity of the Brazilian population results from three ethnic groups admixture: Europeans, Africans and Amerindians, thus increasing the difficulty of performing cystic fibrosis (CF) diagnosis. The nasal potential difference (NPD) evaluates the cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC) activity. Despite being a useful CF diagnostic test and a biomarker of CFTR-modulator drugs, it is also highly operator dependent. Therefore, it may be difficult to get accurate results and to interpret them. Wilschanski and Sermet scores were proposed to address these issues. This study aimed to evaluate repeatability and diagnostic value of NPD parameters and Wilschanski and Sermet scores in a CF center in Rio de Janeiro. NPD was performed in 78 subjects. Maximal PD, amiloride response, total chloride response, and Wilschanski and Sermet scores were explored as means (confidence interval, CI). One-way ANOVA was used to compare mean differences and Scheffe test was used to pair-wise comparisons. Repeatability was evaluated by scatter and Bland-Altman plots. The Ethics Committee of the CF Center has approved the study protocol. Parents and adult participants signed an informed consent form. Forty-eight healthy-volunteers, 19 non-CF and 11 CF patients were enrolled in this study. Significant differences were found when comparing CF patients' NPD parameters to the other two groups (P = 0.000). Moreover, no significant differences were found when parameters from non-CF patients were compared with those from healthy volunteers (P > 0.05). The means of NPD parameters and diagnostic scores of each group were in concordance with disease/non-disease conditions. The repeatability data - Wilschanski and Sermet and NPD - allow NPD to be performed in this Brazilian CF Center. The present study gathered consistent data for Bland-Altman plots. The results of Wilschanski and Sermet diagnostic scores suggest that they were concordant with CF/non-CF conditions. More NPD tests should be performed in the Rio de Janeiro CF dynamic cohort to contribute to international NPD validation studies and to provide NPD as a biomarker in Brazil.


Sequence analysis and distribution in Salmonella enterica serovars of IS3-like elements.
The genome of Salmonella enterica serovar Enteritidis was shown to possess three IS3-like insertion elements, designated IS1230A, B and C, and each was cloned and their respective deoxynucleotide sequences determined. Mutations in elements IS1230A and B resulted in frameshifts in the open reading frames that encoded a putative transposase to be inactive. IS1230C was truncated at nucleotide 774 relative to IS1230B and therefore did not possess the 3' terminal inverted repeat. The three IS1230 derivatives were closely related to each other based on nucleotide sequence similarity. IS1230A was located adjacent to the sef operon encoding SEF14 fimbriae located at minute 97 of the genome of S. Enteritidis. IS1230B was located adjacent to the umuDC operon at minute 42.5 on the genome, itself located near to one terminus of an 815-kb genome inversion of S. Enteritidis relative to S. Typhimurium. IS1230C was located next to attB, the bacteriophage P22 attachment site, and proB, encoding gamma-glutamyl phosphate reductase. A truncated 3' remnant of IS1230, designated IS1230T, was identified in a clinical isolate of S. Typhimurium DT193 strain 2391. This element was located next to attB adjacent to which were bacteriophage P22-like sequences. Southern hybridisation of total genomic DNA from eighteen phage types of S. Enteritidis and eighteen definitive types of S. Typhimurium showed similar, if not identical, restriction fragment profiles in the respective serovars when probed with IS1230A.


Fluoride exposure and dental fluorosis in Newburgh and Kingston, New York: policy implications.
This analysis was conducted to determine the changes in the effect of exposure to fluoridation and other sources of fluoride on dental fluorosis in children attending Newburgh and Kingston school districts in New York State. Data for this analysis were obtained from two surveys conducted in the 1986 and 1995 school years. Analyses were limited to 3500, 7-14-year-old lifelong residents of a fluoridated or a nonfluoridated community. Dean's classification and DMFS index were used for recording dental fluorosis and caries, respectively. A questionnaire was used to collect fluoride exposure data. Regression procedures were used to estimate the effect of fluoridation, fluoride supplements, and brushing before the age of 2 years on dental fluorosis. Children examined in 1996 were at higher risk for both questionable and very mild to severe dental fluorosis if they received fluoride from water or daily tablet use, or started brushing before the age of 2 years. The increase in risk from 1986 to 1995 was greater for African-American children. This analysis showed that the risk of developing dental fluorosis did not decline over time in these communities. Continuous exposure to water fluoridation had an observable effect on dental fluorosis. However, implementation of fluoridation in Newburgh Town did not result in an increase in dental fluorosis prevalence.


Functional and compositional succession of bacterioplankton in response to a gradient in bioavailable dissolved organic carbon.
Studies indicate that bacterial taxa utilize different fractions of the dissolved organic carbon (DOC) pool, while others suggest functional redundancy among constituents of bacterioplankton, implying only a weak coupling between community structure and function. We examined bacterial compositional and functional [ectoenzymatic activities and growth efficiency; bacterial growth efficiency (BGE)] responses to a gradient in bioavailable DOC (bDOC). This was achieved over 10 days in DOC utilization assays containing Baltic Sea water with variable amounts of natural bDOC. Measurements of bacterial growth, O2 and DOC consumption in the assays using non-invasive sampling showed that BGE changed over time and that the bDOC utilized accounted for 4-13% of the DOC pool. Pyrosequencing of 16S rRNA genes demonstrated minor differences at the phylum level between samples, whereas larger successional differences were discernible at lower phylogenetic levels. Our study suggests that changes in concentrations of bDOC affect bacterioplankton BGE and community structure by selecting for some taxa while the relative abundance of most taxa remained unaffected. Ectoenzymes activities suggested preferential degradation of protein-rich compounds by bacteria, switching to carbohydrate-rich DOC when proteins were depleted. Hence, there was a fairly weak linkage between bacterial community composition and DOC utilization suggesting that overall bacterioplankton community structure only to some extent has predictive power for processing of the DOC pool.


Improving diagnostic recognition of primary hyperparathyroidism with machine learning.
Parathyroidectomy offers the only cure for primary hyperparathyroidism, but today only 50% of primary hyperparathyroidism patients are referred for operation, in large part, because the condition is widely under-recognized. The diagnosis of primary hyperparathyroidism can be especially challenging with mild biochemical indices. Machine learning is a collection of methods in which computers build predictive algorithms based on labeled examples. With the aim of facilitating diagnosis, we tested the ability of machine learning to distinguish primary hyperparathyroidism from normal physiology using clinical and laboratory data. This retrospective cohort study used a labeled training set and 10-fold cross-validation to evaluate accuracy of the algorithm. Measures of accuracy included area under the receiver operating characteristic curve, precision (sensitivity), and positive and negative predictive value. Several different algorithms and ensembles of algorithms were tested using the Weka platform. Among 11,830 patients managed operatively at 3 high-volume endocrine surgery programs from March 2001 to August 2013, 6,777 underwent parathyroidectomy for confirmed primary hyperparathyroidism, and 5,053 control patients without primary hyperparathyroidism underwent thyroidectomy. Test-set accuracies for machine learning models were determined using 10-fold cross-validation. Age, sex, and serum levels of preoperative calcium, phosphate, parathyroid hormone, vitamin D, and creatinine were defined as potential predictors of primary hyperparathyroidism. Mild primary hyperparathyroidism was defined as primary hyperparathyroidism with normal preoperative calcium or parathyroid hormone levels. After testing a variety of machine learning algorithms, Bayesian network models proved most accurate, classifying correctly 95.2% of all primary hyperparathyroidism patients (area under receiver operating characteristic = 0.989). Omitting parathyroid hormone from the model did not decrease the accuracy significantly (area under receiver operating characteristic = 0.985). In mild disease cases, however, the Bayesian network model classified correctly 71.1% of patients with normal calcium and 92.1% with normal parathyroid hormone levels preoperatively. Bayesian networking and AdaBoost improved the accuracy of all parathyroid hormone patients to 97.2% cases (area under receiver operating characteristic = 0.994), and 91.9% of primary hyperparathyroidism patients with mild disease. This was significantly improved relative to Bayesian networking alone (P < .0001). Machine learning can diagnose accurately primary hyperparathyroidism without human input even in mild disease. Incorporation of this tool into electronic medical record systems may aid in recognition of this under-diagnosed disorder.


HtrA stress protein is involved in intramacrophagic replication of adherent and invasive Escherichia coli strain LF82 isolated from a patient with Crohn's disease.
Adherent and invasive Escherichia coli (AIEC) bacteria isolated from Crohn's disease patients are able to greatly replicate within macrophages without escaping from the phagosome and without inducing macrophage death. In the present study, evidence is provided that in AIEC strain LF82 the htrA gene encoding the stress protein HtrA is essential for intracellular replication within J774-A1 macrophages. Deletion of the htrA gene in strain LF82 induced increased sensitivity of the isogenic mutant to oxidative stress caused by hydrogen peroxide and a reduced rate of growth in an acid and nutrient-poor medium partly reproducing the microenvironment of the phagosome. In vitro experiments using an LF82 htrA gene promoter fusion with the lacZ gene revealed a 38-fold activation of the promoter in AIEC LF82 intramacrophagic bacteria. The CpxRA two-component signaling pathway was not involved in this activation. In addition, the activation of the LF82 htrA gene promoter was not observed in the nonpathogenic E. coli K-12 intramacrophagic bacteria, indicating that the AIEC LF82 genetic background is crucial for induction of htrA gene transcription during phagocytosis.


Testing strategies to establish the safety of nanomaterials: conclusions of an ECETOC workshop.
The European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) convened a workshop in Barcelona, Spain, in November 2005 to develop testing strategies to establish the safety of nanomaterials. It brought together about 70 scientific and clinical experts from industry, academia, government agencies, research institutes, and nongovernmental organizations. The primary questions to be addressed were the following: What can we do today, and what do we need tomorrow? The three major themes of the workshop were: (1) the need for enhanced efforts in nanomaterial characterization; (2) methodologies for assessments of airborne and internal exposures to nanomaterials; and (3) evaluation of the hazard potential--primarily focusing on pulmonary or dermal routes of exposures. Some of the summary conclusions of the workshop included the following: For the development of nanoparticle characterization, the working definition of nanoparticles was defined as < 100 nm in one dimension or < 1000 nm to include aggregates and agglomerates. Moreover, it was concluded that although many physical factors can influence toxicity, including nanoparticle composition, it is dissolution, surface area and characteristics, size, size distribution, and shape that largely determine the functional, toxicological and environmental impact of nanomaterials. In addition, most of the information on potential systemic effects has thus far been derived from combustion-generated particles. With respect to the assessment of external exposures and metrics appropriate for nanoparticles, the general view of the meeting was that currently it is not possible or desirable to select one form of dose metric (i.e., mass, surface area, or particle number) as the most appropriate measure source. However, it was clear that the surface area metric was likely to be of interest and requires further development. In addition, there is a clear and immediate need to develop instruments which are smaller, more portable, and less expensive than the currently available state of the art instrumentation. With regard to a general testing approach for human health hazard evaluation of nanoparticles, a first step to determine potency may include a prioritization-related in vitro screening strategy to assess the possible reactivity, biomarkers of inflammation and cellular uptake of nanoparticles; however this process should be validated using in vivo techniques. A Tier 1 in vivo testing strategy could include a short-term inhalation or intratracheal instillation of nanoparticles as the route of exposure in the lungs of rats or mice. The endpoints that should be assessed include indices of lung inflammation, cytotoxicity, and cell proliferation, as well as histopathology of the respiratory tract and the major extrapulmonary organs. For Tier 2 in vivo testing for hazard identification, a longer term inhalation study is recommended, and this would include more substantive mechanistic endpoints such as determination of particle deposition, translocation, and disposition within the body. Additional studies could be designed with specific animal models to mimic sensitive populations. With regard to dermal exposures, currently there is little evidence that nanoparticles at a size exceeding 100 nm penetrate through the skin barrier into the living tissue (i.e., dermal compartment). The penetration of nanoparticles at a size less than 100 nm should be a topic of further investigation. Moreover, considering the impacts of dermal exposures and corresponding hazard potential of nanoparticles, it must be taken into consideration that the dermal uptake of nanoparticles will be an order of magnitude smaller than the uptake via the inhalation or oral routes of exposure. For the evaluation of the health risk of nanoparticles, it has to be determined whether they are harmful to living cells and whether, under real conditions, they penetrate through the skin barrier into the living tissue. For the evaluation of the penetration processes, in principle, three methods are available. Using the method of differential stripping, the penetration kinetics of nanoparticles in the stratum corneum and the hair follicles can be evaluated. This analysis can be carried out in vivo. Diffusion cell experiments are an efficient method for in vitro penetration studies. Also, laser scanning microscopy is well suited to test penetration kinetics, although requiring fluorescent-labeled nanoparticles. Emerging topics such as (1) environmental safety testing, (2) applications of nanoparticles for medical purposes, and (3) pathways of inhaled nanoparticles to the central nervous system were also briefly addressed during this workshop. However, it has become clear that these topics should be the subjects of separate workshops and they are not further addressed in this report.


Association of a common dog leucocyte antigen class II haplotype with canine primary immune-mediated haemolytic anaemia.
Immune-mediated haemolytic anaemia (IMHA) is the commonest immune-mediated disease of the dog, representing a major health concern to this species. The aim of this investigation was to determine whether genetic susceptibility to IMHA is associated with genes of the canine major histocompatibility complex (MHC; dog leucocyte antigen system, DLA). Samples were collected from 108 dogs with primary idiopathic, Coombs' positive IMHA. This diseased population was subdivided on the basis of Coombs' test results into two groups: 1) dogs with dominant warm-reactive immunoglobulin (Ig) G haemagglutinins and (2) dogs with an additional or dominant cold-reactive IgM haemagglutinin. The DLA class II alleles and haplotypes of the diseased population were characterised, and these data were compared with those derived from a breed-matched control cohort and a much larger group of DLA-typed dogs. Two haplotypes were increased in the patient group: DLA-DRB1*00601/DQA1*005011/DQB1*00701 (in the group with warm-reactive IgG haemagglutinins only) and DLA-DRB1*015/DQA1*00601/DQB1*00301 (in both groups, but more so in the group with cold-reactive IgM haemagglutinins). One haplotype, DLA-DRB1*001/DQA1*00101/DQB1*00201, was decreased in the total patient group, but this decrease was limited to the warm-reactive IgG haemagglutinins group, and it was actually increased in the cold-reactive IgM haemagglutinins group. A second haplotype, DLA-DRB1*015/DQA1*00601/DQB1*02301, was also decreased in the total patient group, and this decrease was found in both subgroups. In addition, all haplotypes carrying DLA-DRB1*001 were significantly increased in the cold-reactive IgM haemagglutinins group. When the overall patient group was divided on the basis of individual breeds with more than six animals represented, each of the haplotypes could be shown to be implicated in one of the breeds. Thus, it was apparent that different breeds had different MHC associations with canine IMHA, which is similar to the observation that different human ethnic groups can have different HLA associations with the same immune-mediated disease.


Oligomeric bile acid-mediated oral delivery of low molecular weight heparin.
Intestinal transporters are limited to the transport of small molecular substrates. Here, we describe the development of apical sodium-dependent bile acid transporter (ASBT)-targeted high-affinity oligomeric bile acid substrates that mediate the transmembrane transport of low molecular weight heparin (LMWH). Several oligomers of deoxycholic acid (oligoDOCA) were synthesized to investigate the substrate specificity of ASBT. To see the binding of oligoDOCA on the substrate-binding pocket of ASBT, molecular docking was used and the dissociation rate constants (KD) were measured using surface plasmon resonance. The KD for tetrameric DOCA (tetraDOCA) was 50-fold lower than that for monomeric DOCA, because tetraDOCA interacted with several hydrophobic grooves in the substrate-binding pocket of ASBT. The synthesized oligoDOCA compounds were subsequently chemically conjugated to macromolecular LMWH. In vitro, tetraDOCA-conjugated LMWH (LHe-tetraD) had highest selectivity for ASBT during its transport. Orally administered LHe-tetraD showed remarkable systemic anticoagulation activity and high oral bioavailability of 33.5±3.2% and 19.9±2.5% in rats and monkeys, respectively. Notably, LHe-tetraD successfully prevented thrombosis in a rat model of deep vein thrombosis. These results represent a major advancement in ASBT-mediated LMWH delivery and may facilitate administration of many important therapeutic macromolecules through a non-invasive oral route.


Enhancement of photoacoustic tomography by ultrasonic computed tomography based on optical excitation of elements of a full-ring transducer array.
Photoacoustic computed tomography (PACT) is a hybrid technique that combines optical excitation and ultrasonic detection to provide high-resolution images in deep tissues. In the image reconstruction, a constant speed of sound (SOS) is normally assumed. This assumption, however, is often not strictly satisfied in deep tissue imaging, due to acoustic heterogeneities within the object and between the object and the coupling medium. If these heterogeneities are not accounted for, they will cause distortions and artifacts in the reconstructed images. In this Letter, we incorporated ultrasonic computed tomography (USCT), which measures the SOS distribution within the object, into our full-ring array PACT system. Without the need for ultrasonic transmitting electronics, USCT was performed using the same laser beam as for PACT measurement. By scanning the laser beam on the array surface, we can sequentially fire different elements. As a first demonstration of the system, we studied the effect of acoustic heterogeneities on photoacoustic vascular imaging. We verified that constant SOS is a reasonable approximation when the SOS variation is small. When the variation is large, distortion will be observed in the periphery of the object, especially in the tangential direction.


Quality of Life in Different Chronic Diseases and Its Related Factors.
Quality of life (QoL) is an important tool for evaluating the effect of a disease as well as effects of treatment interventions. The present study is aimed to investigate QoL of the patients with various prevalent chronic diseases in Iran. In a cross-sectional study, 625 patients with one of the prevalent chronic diseases were assessed. Participants were recruited via convenient sampling method in special clinics of chronic diseases in three educational hospitals in Qazvin, Iran, from May to December 2016. Data were collected using WHOQOL-BREF and demographic questionnaire through interview. Data were analyzed using independent t-test, ANOVA, Pearson's correlation coefficient, and multiple linear regression method. Significant difference in QoL scores was found in different chronic disease; patients with diabetes and hypertension had higher scores and patients with asthma and chronic obstructive pulmonary disease (COPD) had the lowest scores. Multivariate analysis showed that age, economic status, and type of disease were among the significant variables in predicting the QoL of the patients with chronic diseases. Some patients, such as those with asthma and COPD, had less QoL than other patients and should be prioritized while planning for the promotion of healthcare services. Due to the negative effects of economic status on QoL, the financial support programs should be among the major priorities of the patients' QoL improvement programs.


Detection of β-alanyl aminopeptidase as a biomarker for Pseudomonas aeruginosa in the sputum of patients with cystic fibrosis using exogenous volatile organic compound evolution.
A novel, rapid and sensitive analytical method has been developed and applied to 105 sputum samples from patients with cystic fibrosis, including 5 samples from post-lung transplant patients. This new method is specifically targeted to measure β-alanyl aminopeptidase activity which is characteristic of some important Gram-negative pathogens. Of relevance to this study are Pseudomonas aeruginosa and pathogens of the Burkholderia cepacia complex both of which are commonly associated with respiratory infections as well as increased morbidity and mortality in adult cystic fibrosis patients. The analytical method involves the addition of a novel enzyme substrate (i.e. 3-amino-N-(3-fluorophenyl)propanamide) that interacts with β-alanyl aminopeptidase to generate an exogenous volatile organic compound 3-fluoroaniline (LOD 0.02 μg mL-1; LOQ 0.06 μg mL-1). 3-Fluoroaniline was determined at 20 times above its calculated limit of quantification in the sputum samples by HS-SPME-GC-MS and then the results compared with standard culture methods and bacterial identification using MALDI-TOF-MS. Detection of 3-fluoroaniline was possible after only 8 h incubation of the sputum samples with a 95% success rate; this increased to 100% at 24 h which was well within the typical routine timeframe of 48 h. To our knowledge, this is the first demonstration of detection of P. aeruginosa by use of a custom-designed substrate to liberate a detectable and unique VOC. The very high negative predictive value (100% in this study) means such an assay could be appropriate as a screening technique for patients who are not yet colonized by this pathogen.


Fractional Brownian motion and multivariate-t models for longitudinal biomedical data, with application to CD4 counts in HIV-positive patients.
Longitudinal data are widely analysed using linear mixed models, with 'random slopes' models particularly common. However, when modelling, for example, longitudinal pre-treatment CD4 cell counts in HIV-positive patients, the incorporation of non-stationary stochastic processes such as Brownian motion has been shown to lead to a more biologically plausible model and a substantial improvement in model fit. In this article, we propose two further extensions. Firstly, we propose the addition of a fractional Brownian motion component, and secondly, we generalise the model to follow a multivariate-t distribution. These extensions are biologically plausible, and each demonstrated substantially improved fit on application to example data from the Concerted Action on SeroConversion to AIDS and Death in Europe study. We also propose novel procedures for residual diagnostic plots that allow such models to be assessed. Cohorts of patients were simulated from the previously reported and newly developed models in order to evaluate differences in predictions made for the timing of treatment initiation under different clinical management strategies. A further simulation study was performed to demonstrate the substantial biases in parameter estimates of the mean slope of CD4 decline with time that can occur when random slopes models are applied in the presence of censoring because of treatment initiation, with the degree of bias found to depend strongly on the treatment initiation rule applied. Our findings indicate that researchers should consider more complex and flexible models for the analysis of longitudinal biomarker data, particularly when there are substantial missing data, and that the parameter estimates from random slopes models must be interpreted with caution.


Mental health service use in a representative sample of National Guard soldiers.
With Operation Iraqi Freedom and Operation Enduring Freedom winding down, large numbers of National Guard members have recently returned from active deployment. No prospective, longitudinal studies have examined predictors of mental health service use in a representative sample of National Guard soldiers. This study investigated the prevalence and predictors of mental health service use in a representative sample of National Guard soldiers. A representative sample (N=1,189) of Ohio Army National Guard participants was enrolled. Demographic characteristics, mental health problems, and deployment status in 2009-2010 and mental health service use during the subsequent one-year period (2010-2011) were assessed. Approximately 16% of National Guard members used mental health services during the one-year period. Among those with depression, posttraumatic stress disorder, anxiety, alcohol use disorders, or suicidal ideation, a little over one-third (37%) reported using services in the subsequent 12 months. Among all National Guard soldiers, being female, black, Hispanic, and insured and having comorbid general medical and mental health problems predicted mental health service use. Among those with mental health problems, only black race predicted mental health service use. Results suggest that there is a substantial level of unmet need for mental health treatment among National Guard soldiers. Efforts to identify National Guard soldiers with a need for mental health services and improving access to care might be warranted.


Psychometric properties of the Hare Psychopathy Checklist-Revised (PCL-R) in a representative sample of Canadian federal offenders.
The Hare Psychopathy Checklist-Revised (PCL-R; Hare, 2003) is a commonly used psychological test for assessing traits of psychopathic personality disorder. Despite the abundance of research using the PCL-R, the vast majority of research used samples of convenience rather than systematic methods to minimize sampling bias and maximize the generalizability of findings. This potentially complicates the interpretation of test scores and research findings, including the "norms" for offenders from the United States and Canada included in the PCL-R manual. In the current study, we evaluated the psychometric properties of PCL-R scores for all male offenders admitted to a regional reception center of the Correctional Service of Canada during a 1-year period (n = 375). Because offenders were admitted for assessment prior to institutional classification, they comprise a sample that was heterogeneous with respect to correctional risks and needs yet representative of all offenders in that region of the service. We examined the distribution of PCL-R scores, classical test theory indices of its structural reliability, the factor structure of test items, and the external correlates of test scores. The findings were highly consistent with those typically reported in previous studies. We interpret these results as indicating it is unlikely any sampling limitations of past research using the PCL-R resulted in findings that were, overall, strongly biased or unrepresentative.


Prostate cancer support groups: Canada-based specialists' perspectives.
To understand prostate cancer (PCa) specialists' views about prostate cancer support groups (PCSGs), a volunteer sample of Canada-based PCa specialists (n = 150), including urologists (n = 100), radiation oncologists (n = 40), and medical oncologists (n = 10) were surveyed. The 56-item questionnaire used in this study included six sets of attitudinal items to measure prostate cancer specialists' beliefs about positive and negative influences of PCSGs, reasons for attending PCSGs, the attributes of effective PCSGs, and the value of face-to-face and web-based PCSGs. In addition, an open-ended question was included to invite additional input from participants. Results showed that PCSGs were positively valued, particularly for information sharing, education and psychosocial support. Inclusivity, privacy, and accessibility were identified as potential barriers, and recommendations were made for better marketing PCSGs to increase engagement. Findings suggest prostate cancer specialists highly valued the role and potential benefits of face-to-face PCSGs. Information provision and an educational role were perceived as key benefits. Some concerns were expressed about the ability of web-based PCSGs to effectively engage and educate men who experience prostate cancer.


Intracranial basal cell carcinoma with extensive invasion of the skull base.
Intracranial invasion of basal cell carcinoma is very rare. A case of a 71-year-old male patient is presented with extensive invasion of middle and posterior fossae, with nearly complete destruction of the petrous bone, involving cerebellopontine angle and cranial nerves by recurrent basal cell carcinoma of the auricular region on the right side. The clinical manifestation was cerebrospinal fluid otorrhoea, facial nerve palsy and trigeminal nerve impairment with hypoesthesia. The patient underwent surgery by combined retroauricular and temporal approach to the skull base. Adjuvant radiation of residual tumor and former tumor recurrence region was performed. MRI studies performed annually show no progress of the tumor. Our patient reports a good quality of life without new neurological deficits 6 years after surgery. Attention should be paid to the malignant nature of basal cell carcinoma making follow up care necessary. In indicated cases in which incomplete excision cannot be excepted and risk factors exist, follow up with CT or/and MRI should be performed to evaluate the infiltrative and invasive character of aggressive basal cell carcinoma and to rule out bone or cerebral infiltration.


Synthesis, pharmacology, and molecular modeling studies of semirigid, nicotinic agonists.
Eight nicotinic agonists were synthesized, and their potencies were estimated by contracture of the frog rectus abdominis muscle. The most potent, 1-methyl-4-acetyl-1,2,3,6-tetrahydropyridine methiodide (3b), 50 times as potent as carbamylcholine, served as a template for the rest. Although all of the agonists could easily conform to the putative nicotinic pharmacophore, their potencies spanned a nearly 10,000-fold range. This pharmacophore, therefore, may be necessary but deficient. Computer-assisted molecular modeling studies helped to delineate additional factors that may contribute to potency. The factors are (1) the ground-state conformation, (2) superimposability of the hydrogen bond acceptor and the cationic head onto the template, (3) electrostatic potential at the cationic head and at the hydrogen bond acceptor site, and (4) the presence of a methyl group bonded to the carbon atom that bears the hydrogen bond acceptor. A new program, ARCHEM, was used to calculate and to visualize electrostatic potentials at the van der Waals surfaces of the agonists.