Distinct sensitivities to phosphate deprivation suggest that RGF peptides play disparate roles in Arabidopsis thaliana root development.
Growing agricultural demands in the face of impending inorganic phosphate (Pi) shortages underscore a need for a better understanding of plant development under conditions of Pi deprivation. Pi is an essential nutrient that is a major component of fertilizer. Plants have evolved strategies to improve the acquisition of this nutrient by altering root development under shortage conditions. We show that signaling peptides thought to act redundantly in Arabidopsis thaliana development have distinct functions in response to Pi deprivation. Using microscopy and confocal imaging, roots were analyzed for growth rate and cellular composition. Using expression microarrays, genes influencing development in response to phosphate deprivation were identified. ROOT GROWTH FACTOR1 (RGF1) and RGF2 influenced different aspects of root development under conditions of Pi deprivation. We found that RGF2 influenced the longitudinal growth rate in the primary root in response to Pi deprivation, whereas RGF1 affected circumferential cell number in the root meristem. These data suggest that the mechanisms controlling adaptive development can depend on disparate functions of genes thought to act redundantly, thus elucidating new functions for important developmental regulators.


Effect of nicotine expectancy and nicotine dose reduction on cigarette demand, withdrawal alleviation, and puff topography.
Current FDA plans include proposed nicotine reduction mandates by the end of 2023. Most research on reduced nicotine cigarettes has been dose-blinded, while a mandate would be known to the public. Few laboratory studies have examined specifically how low nicotine content labeling impacts behavioral response. The purpose of this within-subject, balanced-placebo, human laboratory study was to evaluate the main and interactive effects of nicotine dose expectancy and dose reduction on cigarette reinforcement, withdrawal alleviation, and puff topography. Participants who smoke daily (N=21; 9 female) completed one practice and four experimental sessions in which expectancy (labeled "average" versus "very low" nicotine) and nicotine dose (0.80mg versus 0.03mg yield) were manipulated. Participants in acute withdrawal sampled experimental cigarettes followed by withdrawal alleviation and puff topography measures. Cigarette demand was measured using an incentivized purchase task. Analyses evaluated main and interactive effects of expectancy and nicotine dose. Nicotine dose manipulation produced expected physiological effects (e.g., heart rate increases) and both reduced nicotine dose and expectation manipulations reduced perceived nicotine content. Expectation of reduced nicotine alone or in combination with reduced nicotine dose did not alter demand, withdrawal alleviation, or topography. Effective withdrawal alleviation was observed in all conditions. These data inform nicotine regulation policy by suggesting limited compensatory harms caused by reduced nicotine expectations. The minimal acute effects of reduced nicotine expectancy or exposure on demand suggests that reduced nicotine standards are likely to generate their greatest public health benefit through the slowing of newly initiating cigarette smoking.


Phytoreovirus T = 1 core plays critical roles in organizing the outer capsid of T = 13 quasi-equivalence.
The structures of the double-shelled rice dwarf virus and of its single-shell core have been determined by cryoelectron microscopy and image reconstruction. The core carries a prominent density located at each of the icosahedral faces of its T = 1 lattice. These protrusions are formed by outer shell trimers, tightly inserted at the threefold positions of the core. Such configuration of the core may guide the assembly of the outer shell, aided by lateral interactions between its subunits, into a T = 13 lattice. The organization of the phytoreovirus capsid elucidates for the first time a general model for assembling two unique T numbers of quasi-equivalence.


Functional analysis of the Alternaria brassicicola non-ribosomal peptide synthetase gene AbNPS2 reveals a role in conidial cell wall construction.
SUMMARY Alternaria brassicicola is a necrotrophic pathogen causing black spot disease on virtually all cultivated Brassica crops worldwide. In many plant pathosystems fungal secondary metabolites derived from non-ribosomal peptide synthetases (NPSs) are phytotoxic virulence factors or are antibiotics thought to be important for niche competition with other micro-organisms. However, many of the functions of NPS genes and their products are largely unknown. In this study, we investigated the function of one of the A. brassicicola NPS genes, AbNPS2. The predicted amino acid sequence of AbNPS2 showed high sequence similarity with A. brassicae, AbrePsy1, Cochliobolus heterostrophus, NPS4 and a Stagonospora nodorum NPS. The AbNPS2 open reading frame was predicted to be 22 kb in length and encodes a large protein (7195 amino acids) showing typical NPS modular organization. Gene expression analysis of AbNPS2 in wild-type fungus indicated that it is expressed almost exclusively in conidia and conidiophores, broadly in the reproductive developmental phase. AbNPS2 gene disruption mutants showed abnormal spore cell wall morphology and a decreased hydrophobicity phenotype. Conidia of abnps2 mutants displayed an aberrantly inflated cell wall and an increase in lipid bodies compared with wild-type. Further phenotypic analyses of abnps2 mutants showed decreased spore germination rates both in vitro and in vivo, and a marked reduction in sporulation in vivo compared with wild-type fungus. Moreover, virulence tests on Brassicas with abnps2 mutants revealed a significant reduction in lesion size compared with wild-type but only when aged spores were used in experiments. Collectively, these results indicate that AbNPS2 plays an important role in development and virulence.


Material characterization of porcine lenticular soluble proteins.
The soluble proteins present in the ocular lens impart important optical and dynamic mechanical properties on the lens. The short-range order of crystallin proteins grants transparency to a very concentrated protein solution. This unique protein system directly enables proper visual function of the eye. These proteins were investigated in steady and oscillatory shear. Steady shear data were fitted with a modified Herschel-Bulkley yield stress model that allows for a Newtonian plateau at low shear rates. The Cox-Merz rule was used in conjunction with large amplitude oscillatory shear to give insight into the degradation of the fluid structure with increasing strain. The shear thinning viscoelastic behavior of these proteins gives rise to beneficial mechanical properties and results from the same short-range order granting optical transparency.


[Comparative study of Parkinson's disease and dementia with Lewy bodies].
Comparative investigation of motor and neuropsychological functions was conducted in 17 patients with clinically established diagnosis of dementia with Lewy bodies (DLB), 21 patients with Parkinson's disease (PD) without dementia and 26 patients with dementia (PDD). No significant differences were found in overall severity of parkinsonian features. However, comparing to PD, patients with DLB rarely had resting tremor, bilateral parkinsonism onset and good response to levadopa medication but more frequently exhibited gaze up palsy and myoclonus. Patients with PDD had more prominent akinesia, rigidity and axial disturbances, as compared to the PD patients without dementia, but there were no significant differences in these variables between patients with DLB and PDD. Comparing to PDD patients, those with DLB poorly performed on the tests measuring attention, verbal fluency and visual-spatial functions. Behavioral disturbances (especially apathy, aspontaneity, euphoria, obsessive-compulsive syndrome, dysinhibition, environment dependence) were severer in the patients with DLB and PDD, but no significant differences were found between these two groups. All the patients with DLB, 14 patients with PDD (53.8%) and 2 patients without dementia (7.4%) had psychotic disorders, which emerged later and were less pronounced in the patients with DLB compared to PDD patients. However, no significant differences were found in motor and neuropsychological impairment between DLB and PDD with psychotic disorders. The results may indicate a typological similarity of clinical manifestations of DLB and PDD that suggest nosologic proximity of these conditions included to synucleinopathies group.


Laccases in Food Industry: Bioprocessing, Potential Industrial and Biotechnological Applications.
Laccase is a multi-copper oxidase that catalyzes the oxidation of one electron of a wide range of phenolic compounds. The enzyme is considered eco-friendly because it requires molecular oxygen as co-substrate for the catalysis and it yields water as the sole by-product. Laccase is commonly produced by fungi but also by some bacteria, insects and plants. Due it is capable of using a wide variety of phenolic and non-phenolic substrates, laccase has potential applications in the food, pharmaceutical and environmental industries; in addition, it has been used since many years in the bleaching of paper pulp. Fungal laccases are mainly extracellular enzyme that can be recovered from the residual compost of industrial production of edible mushrooms as Agaricus bisporus and Pleurotus ostreatus. It has also been isolated from microorganisms present in wastewater. The great potential of laccase lies in its ability to oxidize lignin, one component of lignocellulosic materials, this feature can be widely exploited on the pretreatment for agro-food wastes valorization. Laccase is one of the enzymes that fits very well in the circular economy concept, this concept has more benefits over linear economy; based on "reduce-reuse-recycle" theory. Currently, biorefinery processes are booming due to the need to generate clean biofuels that do not come from oil. In that sense, laccase is capable of degrading lignocellulosic materials that serve as raw material in these processes, so the enzyme's potential is evident. This review will critically describe the production sources of laccase as by-product from food industry, bioprocessing of food industry by-products using laccase, and its application in food industry.


The complete amino acid sequence of a mouse kappa light chain.
The complete amino acid sequence of the kappa-chain of the mouse myeloma protein MOPC 21 was established. The protein was reduced and alkylated with iodo[2-(14)C]acetic acid, and 21 tryptic peptides were isolated, mainly by paper electrophoresis and paper chromatography. Three large tryptic peptides (of 35, 36 and 42 residues), which were difficult to isolate in this manner, were obtained pure and in excellent yields by a combination of Sephadex G-50 gel filtration in 1% (w/v) NH(4)HCO(3) and chromatography on a DEAE-cellulose column in ammonium acetate buffer, pH8.1. Peptides overlapping the tryptic peptides were isolated from a chymotryptic digest. The chain is 214 residues long. Microheterogeneity of two peptides was observed and is believed to be due to deamidation. It was not excluded that such deamidation could occur in serum from which the protein was isolated. The sequence is compared with the sequences of two other mouse kappa-chains, and with the human kappa-chain basic sequences.


The Invs gene encodes a microtubule-associated protein.
Microtubule networks are important for many vital processes such as mitosis, cell polarity, and differentiation. Ciliary architecture and function closely depend on the microtubule cytoskeleton, and recent studies suggest a role of apical cilia of renal epithelia in the pathogenesis of polycystic kidney disease. This study evaluates the localization, potential interacting partners, and functional aspects of the Invs gene product inversin. Only recently, INVS has been identified as the gene that is mutated in nephronophthisis type 2, an autosomal recessive polycystic kidney disease. Using immunoprecipitation and co-pelleting assays, we show that the Invs gene product inversin forms a stable complex with tubulin in cultured renal epithelial cells. Inversin localizes to several components of the cytoskeleton including ciliary, random, and polarized microtubule pools. During cell divison, inversin is recruited to mitotic spindle fibers. After microtubule depolymerization using colcemid inversin and tubulin staining is no longer characterized by a network pattern but by homogeneous, diffuse distribution. Inversin does not coprecipitate with tubulin after addition of colcemid. After removal of colcemid, inversin immunofluorescence reappears together with tubulin in centrioles. Treatment with the microtubule stabilizing agent paclitaxel leads to severe alteration of the microtubule cytoskeleton with bundling and formation of long spindles of tubulin and inversin. In conclusion, inversin is closely associated with the microtubule cytoskeleton, and its spatial distribution is dependent on tubulin polymerization. Hence, altered inversin-tubulin interaction may impair ciliary function and thereby contribute to cyst development in nephronophthisis.


Up-regulation of G-protein-coupled receptors for endothelin and thromboxane by lipid-soluble smoke particles in renal artery of rat.
Up-regulation of G-protein-coupled receptors (GPCR) plays key roles in renal hypertension and cardiovascular disease pathogenesis. The present study was designed to examine if lipid-soluble cigarette smoking particles (DSP), nicotine and endotoxin (LPS), induce GPCR up-regulation for thromboxane A(2) (TP), endothelin type A (ET(A) ) and type B (ET(B) ) receptors in renal artery, and if intracellular signal mechanisms are involved. Renal artery segments of rats were exposed to DSP, nicotine or LPS, in organ culture for up to 24 hr. The GPCR-mediated contractions were recorded by using a myograph system. Expression of the GPCR was examined by real-time PCR and immunohistochemistry at mRNA and protein levels. Sarafatoxin 6c (S6c, selective ET(B) receptor agonist), endothelin-1 (ET-1, non-selective ET(A) and ET(B) receptor agonist) and 9,11-Dideoxy-9a,11a-methanoepoxy prostaglandin F(2a) (U46619, a TP receptor agonist) induced contractions were significantly increased after the arterial segments exposed to DSP in a concentration-dependent (0.1-0.4 μl/ml) manner, and S6c also induced a time-dependent contraction, compared to control (dimethyl sulfoxide). This was in parallel with enhanced mRNA expression for ET(B) receptor but not ET(A) and TP receptors, while increased protein expression for ET(A) , ET(B) and TP receptors was seen. The specific nuclear factor-kappa B (NF-κB) signal pathway inhibitor BMS345541 was applied to link DSP effects to the GPCR up-regulation. It totally abolished ET(B) receptor up-regulation, but not ET(A) and TP receptor up-regulations. Our results suggest that DSP transcriptionally up-regulated ET(B) receptor expression in rat renal artery via NF-κB signal pathways, whereas up-regulation of ET(A) and TP receptor-mediated contraction may involve post-transcriptional mechanisms.


[Sleep disturbances in children and adolescents with psychiatric disorders - affective and anxiety disorders].
Sleep disorders are common in children and adolescents, their incidence is estimated to be within 25-40%. Among children with psychiatric disorders, sleep problems often arise. Sleep disorders are a symptom of many mental illnesses, they have a major impact on the severity of other symptoms of the disease and the treatment is often difficult. Epidemiological and clinical studies have shown that children in this group often suffer from chronic insomnia, but also relatively frequently are seen in them primary sleep disorders such as restless legs syndrome or obstructive sleep apnea. On the other hand, there is evidence that primary insomnia may be a risk factor for many psychiatric diseases especially depression and anxiety disturbances, and may even produce symptoms of mental illness. Researches regarding pharmacotherapy in pediatric sleep disorders are limited. As a first-line treatment nonpharmacologic strategies are usually recommended.


Sleep and childhood maltreatment: A matched-control study of sleep characteristics in Norwegian 16-19-year-olds.
Sleep problems are an important but understudied health problem in adolescents exposed to childhood maltreatment. The current study aimed to examine sleep characteristics and insomnia in a population with maltreatment history and compare them to a general population sample of adolescents. Data from a sample of Norwegian 16-19-year-old adolescents with childhood maltreatment experiences (n = 62) were linked to an age- and sex-matched general population sample (n = 238). Independent samples t tests were used to compare the two groups on several detailed sleep parameters, while general linear models were used to control for mental health problems, assessed by the Strengths and Difficulties Questionnaire. Mental health was examined as a potential mediator between childhood maltreatment and sleep characteristics. Adolescents exposed to childhood maltreatment had longer sleep onset latency (1:15 h vs. 47 min, p < 0.001), longer wake after sleep onset (34 min vs. 15 min, p < 0.003), lower sleep efficiency in weekdays (77.8%, vs. 85%, p < 0.007), and lower sleep efficiency in weekends (81.8% vs. 88.9%, p < 0.001) than the reference group. Mental health problems fully mediated the association between childhood maltreatment exposure and longer sleep onset latency and wake after sleep onset. The rate of insomnia did not significantly differ between the groups. This study suggests that older adolescents exposed to childhood maltreatment have more sleep problems than the general adolescent population, and that these problems may be explained by co-existing mental health problems. These findings highlight the potential utility of assessing and providing sleep-focused treatment to adolescents with a history of childhood maltreatment and mental health problems.


Potential markers of aggressiveness in prostatic intraepithelial neoplasia detected by fluorescence in situ hybridization.
High-grade prostatic intraepithelial neoplasia (PIN) represents the most likely precursor of prostatic adenocarcinoma. In this review, we discuss the utility of different techniques of interphase fluorescence in situ hybridization (FISH) in evaluating the genetic association between PIN and adenocarcinoma. Although the proportion of PIN and prostatic carcinoma foci that have chromosomal anomalies is similar, foci of carcinoma often have more alterations. This supports the hypothesis that PIN is a precursor of carcinoma. In some prostates, however, PIN foci have more alterations than matched carcinoma foci, suggesting that PIN foci can sometimes undergo more extensive chromosome evolution than carcinoma foci. Gain of chromosome 8 is the most common numerical alteration in prostatic carcinoma and PIN foci, and is associated with increasing cancer stage and grade. Thus, genes on chromosome 8 may play a role in the initiation and progression of prostatic carcinoma. One primary tumor focus usually shares chromosomal anomalies with associated lymph node metastases, implying that only one primary lesion acquires genetic alterations that allow it to escape the prostate. PIN and prostatic carcinoma have similar genetic changes. This supports the hypothesis that PIN is often a precursor of carcinoma. Genes on chromosome 8 may play a role in both initiation and progression of prostatic carcinoma.


A Survey on Voiding Complaints in Women Presenting at a Pelvic Care Center.
This article reports the prevalence of self-reported voiding complaints and the relationship with other pelvic floor and bladder dysfunctions (PFD). Women with a variety of PFDs were referred to the pelvic care center. A standardised questionnaire on 6 PFDs was used. Frequencies of patient characteristics, PFDs and voiding complaints were calculated. Cross tabulation was used to investigate correlations and Pearson correlation coefficients to reveal the strength of the association between PFDs and self-reported voiding complaints. Data of 4470 women were included. Prevalence of (self-reported) voiding Lower urinary tract symptoms was 59.5%. Incomplete bladder emptying is the most prevalent voiding complaint. Self-reported voiding complaints are weakly correlated to age (r = 0.15, p < 0.01) and have moderate correlation with self-reported recurrent urinary tract infections (r = 0.34, p < 0.01), pelvic floor, bladder and bowel complaints. However, the correlation between the feeling of incomplete bladder emptying and the presence of recurrent urinary tract infections is weak (r = 0.06, p = 0.02). Voiding complaints have a high prevalence and symptom bother in women visiting a pelvic care center.


Dual signal transduction through delta opioid receptors in a transfected human T-cell line.
Opiates are known to function as immunomodulators, in part by effects on T cells. However, the signal transduction pathways mediating the effects of opiates on T cells are largely undefined. To determine whether pathways that regulate free intracellular calcium ([Ca2+]i) and/or cAMP are affected by opiates acting through delta-type opioid receptors (DORs), a cDNA encoding the neuronal DOR was expressed in a stably transfected Jurkat T-cell line. The DOR agonists, deltorphin and [D-Ala2, D-Leu5]-enkephalin (DADLE), elevated [Ca2+]i, measured by flow cytofluorometry using the calcium-sensitive dye, Fluo-3. At concentrations from 10(-11)-10(-7) M, both agonists increased [Ca2+]i from 60 nM to peak concentrations of 400 nM in a dose-dependent manner within 30 sec (ED50 of approximately 5 x 10(-9) M). Naltrindole, a selective DOR antagonist, abolished the increase in [Ca2+]i, and pretreatment with pertussis toxin was also effective. To assess the role of extracellular calcium, cells were pretreated with EGTA, which reduced the initial deltorphin-induced elevation of [Ca2+]i by more than 50% and eliminated the second phase of calcium mobilization. Additionally, the effect of DADLE on forskolin-stimulated cAMP production was determined. DADLE reduced cAMP production by 70% (IC50 of approximately equal to 10(-11) M), and pertussis toxin inhibited the action of DADLE. Thus, the DOR expressed by a transfected Jurkat T-cell line is positively coupled to pathways leading to calcium mobilization and negatively coupled to adenylate cyclase. These studies identify two pertussis toxin-sensitive, G protein-mediated signaling pathways through which DOR agonists regulate the levels of intracellular messengers that modulate T-cell activation.


Inhibitor of cyclin-dependent kinase (CDK) interacting with cyclin A1 (INCA1) regulates proliferation and is repressed by oncogenic signaling.
The cell cycle is driven by the kinase activity of cyclin·cyclin-dependent kinase (CDK) complexes, which is negatively regulated by CDK inhibitor proteins. Recently, we identified INCA1 as an interaction partner and a substrate of cyclin A1 in complex with CDK2. On a functional level, we identified a novel cyclin-binding site in the INCA1 protein. INCA1 inhibited CDK2 activity and cell proliferation. The inhibitory effects depended on the cyclin-interacting domain. Mitogenic and oncogenic signals suppressed INCA1 expression, whereas it was induced by cell cycle arrest. We established a deletional mouse model that showed increased CDK2 activity in spleen with altered spleen architecture in Inca1(-/-) mice. Inca1(-/-) embryonic fibroblasts showed an increase in the fraction of S-phase cells. Furthermore, blasts from acute lymphoid leukemia and acute myeloid leukemia patients expressed significantly reduced INCA1 levels highlighting its relevance for growth control in vivo. Taken together, this study identifies a novel CDK inhibitor with reduced expression in acute myeloid and lymphoid leukemia. The molecular events that control the cell cycle occur in a sequential process to ensure a tight regulation, which is important for the survival of a cell and includes the detection and repair of genetic damage and the prevention of uncontrolled cell division.


RNA- and ATAC-sequencing Reveals a Unique CD83+ Microglial Population Focally Depleted in Parkinson's Disease.
All brain areas affected in Parkinson's disease (PD) show an abundance of microglia with an activated morphology together with increased expression of pro-inflammatory cytokines, suggesting that neuroinflammation may contribute to the neurodegenerative process in this common and incurable disorder. We applied a single nucleus RNA- and ATAC-sequencing approach using the 10x Genomics Chromium platform to postmortem PD samples to investigate microglial heterogeneity in PD. We created a multiomic dataset using substantia nigra (SN) tissues from 19 PD donors and 14 non-PD controls (NPCs), as well as three other brain regions from the PD donors which are differentially affected in this disease: the ventral tegmental area (VTA), substantia inominata (SI), and hypothalamus (HypoTs). We identified thirteen microglial subpopulations within these tissues as well as a perivascular macrophage and a monocyte population, of which we characterized the transcriptional and chromatin repertoires. Using this data, we investigated whether these microglial subpopulations have any association with PD and whether they have regional specificity. We uncovered several changes in microglial subpopulations in PD, which appear to parallel the magnitude of neurodegeneration across these four selected brain regions. Specifically, we identified that inflammatory microglia in PD are more prevalent in the SN and differentially express PD-associated markers. Our analysis revealed the depletion of a CD83 and HIF1A- expressing microglial subpopulation, specifically in the SN in PD, that has a unique chromatin signature compared to other microglial subpopulations. Interestingly, this microglial subpopulation has regional specificity to the brainstem in non-disease tissues. Furthermore, it is highly enriched for transcripts of proteins involved in antigen presentation and heat-shock proteins, and its depletion in the PD SN may have implications for neuronal vulnerability in disease.


CYP2D6 catalyzes tamoxifen 4-hydroxylation in human liver.
The major metabolites of tamoxifen (tam) formed by animal and human liver microsomes are mono-N-demethylated tam, 4-hydroxy-tam (4-OH-tam), and tam-N-oxide. The N-desmethylated-tam and 4-OH-tam are formed by P450s, whereas the N-oxide is primarily formed by flavin-containing monooxygenase. Because 4-OH-tam is a highly potent antiestrogen (and possibly is the active anticancer tam metabolite) and is on the path of formation of the reactive intermediate that binds covalently to proteins and DNA, it was of importance to identify the P450(s) catalyzing its formation. In the current study, three different preparations of expressed human P450s in Escherichia coli, lymphoblastoma cells, and insect cell line and livers from several human donors were used to identify the P450 isoform catalyzing the 4-hydroxylation (preliminary results were reported by Dehal et al., Eleventh International Symposium on Microsomes and Drug Oxidations, p. 71. Los Angeles, 1996). Tam metabolism was examined with human CYP2C8, 2C9, 2C18, 2C19, and 2D6 expressed in E. coli. These P450s were reconstituted with P450 reductase and lipid and were incubated with 50 microM [3H]tam and NADPH at 37 degrees C for 60 min. Essentially all of the recombinant P450s catalyzed the N-demethylation to various degrees; however, only 2D6 yielded detectable levels of 4-OH-tam. The inclusion of cytochrome b5 in the reconstituted system of 2D6 and 2C9 did not significantly affect the rate of 4-hydroxylation, indicating that b5 is not essential for this activity. Tam metabolism by CYP1A1, 1A2, 2A6, 2B6, 2C8, 2C9, 2D6, 2E1, and 3A4, expressed in lymphoblastoma cells, revealed that only 2D6 significantly catalyzed the 4-hydroxylation. Tam metabolism by CYP2D6 coexpressed with P450 reductase in a baculovirus infected insect cell line ("supersomes") exhibited marked tam 4-hydroxylation. In an experiment with human liver microsomes, the inclusion of quinidine, a specific 2D6 inhibitor, resulted in approximately 50% inhibition of tam 4-hydroxylation without affecting N-demethylation. Polyclonal antibodies raised against 2D6 moderately inhibited (approximately 30%) the 4-hydroxylation in human liver microsomes. These results demonstrate a significant contribution by CYP2D6 to the catalysis of tam-4-hydroxylation by human liver.


Development of a dynamic water budget model for Abu Dhabi Emirate, UAE.
In this study, a dynamic water budget model is developed for the Emirate of Abu Dhabi (EAD) in the United Arab Emirates (UAE). The model, called Abu Dhabi Water Budget Model (ADWBM), accounts for a number of drivers such as population growth, economic growth, consumption pattern and climatic factors. Model formulation, calibration, validation as well as simulation results for two future situations are presented in this paper. The two water simulations discuss demand-side options in response to different future water conditions until 2050. The first simulation, namely, baseline (BL) simulation examined water balance in the emirate assuming no change in both water production and consumption. BL simulation results highlight the expected shortages in water resources assuming no modification in the supply side. The second simulation, a more conservative and practical simulation considering water conservation options and sustainable improvements to the supply side was developed to achieve a balanced water budget by reducing the baseline consumption rates. The results show that a significant demand reduction is needed in all demand sectors, reaching 60% in the potable sectors and above 70% in non-potable sectors. Overall, results show that the ADWBM can be used as a numerical tool to produce accurate figures of water supply and demand for the sake of planning and decision making in the water sector of the EAD until 2050.


Synthesis and Antimicrobial Evaluation of Amixicile-Based Inhibitors of the Pyruvate-Ferredoxin Oxidoreductases of Anaerobic Bacteria and Epsilonproteobacteria.
Amixicile is a promising derivative of nitazoxanide (an antiparasitic therapeutic) developed to treat systemic infections caused by anaerobic bacteria, anaerobic parasites, and members of the Epsilonproteobacteria (Campylobacter and Helicobacter). Amixicile selectively inhibits pyruvate-ferredoxin oxidoreductase (PFOR) and related enzymes by inhibiting the function of the vitamin B1 cofactor (thiamine pyrophosphate) by a novel mechanism. Here, we interrogate the amixicile scaffold, guided by docking simulations, direct PFOR inhibition assays, and MIC tests against Clostridium difficile, Campylobacter jejuni, and Helicobacter pylori Docking simulations revealed that the nitro group present in nitazoxanide interacts with the protonated N4'-aminopyrimidine of thiamine pyrophosphate (TPP). The ortho-propylamine on the benzene ring formed an electrostatic interaction with an aspartic acid moiety (B456) of PFOR that correlated with improved PFOR-inhibitory activity and potency by MIC tests. Aryl substitution with electron-withdrawing groups and substitutions of the propylamine with other alkyl amines or nitrogen-containing heterocycles both improved PFOR inhibition and, in many cases, biological activity against C. difficile Docking simulation results correlate well with mechanistic enzymology and nuclear magnetic resonance (NMR) studies that show members of this class of antimicrobials to be specific inhibitors of vitamin B1 function by proton abstraction, which is both novel and likely to limit mutation-based drug resistance.


Heat-induced symmetry breaking in ant (Hymenoptera: Formicidae) escape behavior.
The collective egress of social insects is important in dangerous situations such as natural disasters or enemy attacks. Some studies have described the phenomenon of symmetry breaking in ants, with two exits induced by a repellent. However, whether symmetry breaking occurs under high temperature conditions, which are a common abiotic stress, remains unknown. In our study, we deposited a group of Polyrhachis dives ants on a heated platform and counted the number of escaping ants with two identical exits. We discovered that ants asymmetrically escaped through two exits when the temperature of the heated platform was >32.75°C. The degree of asymmetry increased linearly with the temperature of the platform. Furthermore, the higher the temperature of heated platform was, the more ants escaped from the heated platform. However, the number of escaping ants decreased for 3 min when the temperature was higher than the critical thermal limit (39.46°C), which is the threshold for ants to endure high temperature without a loss of performance. Moreover, the ants tended to form small groups to escape from the thermal stress. A preparatory formation of ant grouping was observed before they reached the exit, indicating that the ants actively clustered rather than accidentally gathered at the exits to escape. We suggest that a combination of individual and grouping ants may help to optimize the likelihood of survival during evacuation.


Next-Generation Sequencing Somatic and Germline Assay Troubleshooting Guide Derived From Proficiency Testing Data.
Next-generation sequencing-based assays are increasingly used in clinical molecular laboratories to detect somatic variants in solid tumors and hematologic malignancies and to detect constitutional variants. Proficiency testing data are potential sources of information about challenges in performing these assays. To examine the most common sources of unacceptable results from the College of American Pathologists Next-Generation Sequencing Bioinformatics, Hematological Malignancies, Solid Tumor, and Germline surveys and provide recommendations on how to avoid these pitfalls and improve performance. The College of American Pathologists next-generation sequencing somatic and germline proficiency testing survey results from 2016 to 2019 were analyzed to identify the most common causes of unacceptable results. On somatic and germline proficiency testing surveys, 95.9% (18 815/19 623) and 97.8% (33 890/34 641) of all variants were correctly identified, respectively. The most common causes of unacceptable results related to sequencing were false-negative errors in genomic regions that were difficult to sequence because of high GC content. False-positive errors occurred in the context of homopolymers and pseudogenes. Recurrent errors in variant annotation were seen for dinucleotide and duplication variants and included unacceptable transcript selection and outdated variant nomenclature. A small percentage of preanalytic or postanalytic errors were attributed to specimen swaps and transcription errors. Laboratories demonstrate overall excellent performance for detecting variants in both somatic and germline proficiency testing surveys. Proficiency testing survey results highlight infrequent, but recurrent, analytic and nonanalytic challenges in performing next- generation sequencing-based assays and point to remedies to help laboratories improve performance.


Guidelines for Clinical Target Volume Definition for Perineural Spread of Major Salivary Gland Cancers.
Postoperative radiotherapy is the standard of care for resected major salivary cancers that are at risk of locoregional recurrence. Of the various histological subtypes, perineural invasion is most common in adenoidcystic carcinomas of the three major salivary glands - parotid, submandibular and sublingual. The clinical target volume (CTV) for these cases must include the relevant cranial nerve pathways at risk. A contouring atlas was devised for delineation of the CTV of the nerves supplying the major salivary glands. Using standard anatomy texts and e-anatomy sources the nerves supplying the major salivary glands were identified. Subsequently the pathways of the nerves were drawn on an archived patient's planning computed tomography scan. The innervation of the major salivary glands has been identified and studied. Both bone and soft tissue CTVs have been delineated. A full set of images and CTVs of all the relevant transverse computed tomography slices has been archived, a number of which are printed in this article. Variation in CTV delineation is a recognised problem in a variety of anatomic sites. Guidelines and atlases can standardise practice and may improve the safety and efficacy of therapy. An atlas has been generated to guide clinicians in delineating the CTVs for perineural spread in major salivary gland cancers.


["Activate your Wait" project : pilot study carried out at a thermal centre]
Physical activity is recognised as a major health determinant, with positive effects on health, environmental sustainability and economy. National surveillance data show that one out of three Italians - adult and elderly alike - declares to be sedentary, with a progressively increasing trend. From the urgent need to implement strategies to promote physical activity the "Activate your Wait" ("Attiva l'Attesa") project was born, aimed at transforming the waiting pauses during day to day life into opportunities to perform simple stretching and active mobilization exercises. The pilot study was carried out at the Terme S. Egidio, Suio Terme Castelforte (Latina). The results of the questionnaire distributed in the pre-intervention phase, aimed at assessing the users' interest in the project and physical activity in general, are reported. The questionnaire was administrated in the waiting rooms in September 2018 and September 2019. A total of 129 subjects responded to the questionnaire: 43% declared themselves sedentary, 73% reported willingness to perform simple physical exercises while waiting, and 76% believed that physical activity during waiting moments could have a positive impact on health. The project's subsequent goal is to identify suitable exercises to be proposed during the intervention period, which can be easily reproduced independently by users in their everyday life.


Evaluation of chromosome 12 copy number in ovarian granulosa cell tumors using interphase cytogenetics.
Trisomy 12 is frequently observed in karyotypes of ovarian sex cord-stromal tumors, including adult and juvenile granulosa cell tumors (AGCTs and JGCTs). We assessed the ability to detect this abnormality in deparaffinized tissue sections of 19 ovarian GCTs (17 AGCTs, two JGCTs) and in one fibrothecoma by simultaneous in situ hybridization with fluorescent-labeled centromeric probes to chromosomes 12 (fluorescein isothiocyanate conjugated) and 17 (rhodamine conjugated). In order to quantitate the artifact introduced by nuclear slicing, such analyses were performed both on intact tissue sections and on cytospins of nuclei prepared by enzymatic dissociation from the corresponding tissue block. The series was also evaluated for numerical abnormalities of chromosome X, a less common cytogenetic finding in GCT. Twelve of 19 cases (63%) displayed evidence of trisomy 12 (defined as signal gain in > or = 10% of nuclei) in the intact section, the cytospin, or both. Trisomy for chromosome 17 was present in one case, and trisomy X was present in two cases. In tissue sections the incidence of signal gain for the chromosome 12 probe varied from 0-45% of nuclei (mean 19%). In cytospin preparations, the percentage signal gain for chromosome 12 ranged from 0 to 84% (mean 33%). This study supports the presence of trisomy 12 as a common, but not defining, cytogenetic anomaly in ovarian GCTs. Its presence, however, within only a minority of tumor cells may be partly explained by slicing artifact associated with intact tissue sections, although partial involvement of intact nuclei suggests that trisomy 12 may also be encountered as a heterogeneous abnormality within neoplastic populations of GCTs.


Allele and genotype frequencies for Primary Hereditary Cataract, Multifocal Retinopathy 1, and Degenerative Myelopathy in Pyrenean Mountain Dog from Italy.
Pyrenean Mountain Dog (PMD) is an ancient dog breed firstly described in XIV century in the Pyrenees Region and nowadays diffused both in Europe and in the US. Hereditary Cataract (HC), defined as the inherited opacity of the lens, involves clinical signs ranging from reduced vision to glaucoma. A molecular basis of HC was firstly described in Staffordshire Bull Terriers and then reported in multiple canine breeds. The HC-associated variation is a single nucleotide deletion in HSF4 gene that introduces a premature stop codon (c.962del, p.Ala321*). Multifocal Retinopathy 1 (MR) is an ocular disorder characterized by multiple areas of retinal degeneration, caused in various dog breeds (including PMD) by a single nucleotide variant (SNV) in BEST1 gene that generates a premature stop codon (c.73G>A, p.Arg25*). Degenerative Myelopathy (DM) is an adult-onset, progressive neurodegenerative disease and it is associated to a SNV in SOD1 gene causing a change in aminoacidic sequence of the protein (c.118G>A, p.Glu40Lys). This causative variant has been described in various dog breeds, including PMD. Aim of this study was to determine the allele frequencies for the abovementioned three genetic diseases in the Italian breeding PMD population. The survey found no dogs carrying the allele (deletion) associated with HC, while three dogs (6%) were heterozygous (G/A) for the MR-associated variant, and seven dogs (13%) were heterozygous (G/A) for the DM-associated alteration, indicating that the variant alleles frequency were 0%, 3%, and 7%, respectively. Appropriate mating management is suggested for the prevention of genetic diseases spreading in the PMD population.


Application of the deflection criterion to classification of radar SAR images.
An original application of the weighted deflection is proposed for radar target classification by quadratic filters. An explicit formulation of optimal filters is derived. We analyse the impact of the weighting parameter on real data recognition, and show that performances are better when the deflection coincides with the Fisher ratio.


Shear-induced endothelial mechanotransduction: the interplay between reactive oxygen species (ROS) and nitric oxide (NO) and the pathophysiological implications.
Hemodynamic shear stress, the blood flow-generated frictional force acting on the vascular endothelial cells, is essential for endothelial homeostasis under normal physiological conditions. Mechanosensors on endothelial cells detect shear stress and transduce it into biochemical signals to trigger vascular adaptive responses. Among the various shear-induced signaling molecules, reactive oxygen species (ROS) and nitric oxide (NO) have been implicated in vascular homeostasis and diseases. In this review, we explore the molecular, cellular, and vascular processes arising from shear-induced signaling (mechanotransduction) with emphasis on the roles of ROS and NO, and also discuss the mechanisms that may lead to excessive vascular remodeling and thus drive pathobiologic processes responsible for atherosclerosis. Current evidence suggests that NADPH oxidase is one of main cellular sources of ROS generation in endothelial cells under flow condition. Flow patterns and magnitude of shear determine the amount of ROS produced by endothelial cells, usually an irregular flow pattern (disturbed or oscillatory) producing higher levels of ROS than a regular flow pattern (steady or pulsatile). ROS production is closely linked to NO generation and elevated levels of ROS lead to low NO bioavailability, as is often observed in endothelial cells exposed to irregular flow. The low NO bioavailability is partly caused by the reaction of ROS with NO to form peroxynitrite, a key molecule which may initiate many pro-atherogenic events. This differential production of ROS and RNS (reactive nitrogen species) under various flow patterns and conditions modulates endothelial gene expression and thus results in differential vascular responses. Moreover, ROS/RNS are able to promote specific post-translational modifications in regulatory proteins (including S-glutathionylation, S-nitrosylation and tyrosine nitration), which constitute chemical signals that are relevant in cardiovascular pathophysiology. Overall, the dynamic interplay between local hemodynamic milieu and the resulting oxidative and S-nitrosative modification of regulatory proteins is important for ensuing vascular homeostasis. Based on available evidence, it is proposed that a regular flow pattern produces lower levels of ROS and higher NO bioavailability, creating an anti-atherogenic environment. On the other hand, an irregular flow pattern results in higher levels of ROS and yet lower NO bioavailability, thus triggering pro-atherogenic effects.


90Y SPECT scatter estimation and voxel dosimetry in radioembolization using a unified deep learning framework.
90Y SPECT-based dosimetry following radioembolization (RE) in liver malignancies is challenging due to the inherent scatter and the poor spatial resolution of bremsstrahlung SPECT. This study explores a deep-learning-based absorbed dose-rate estimation method for 90Y that mitigates the impact of poor SPECT image quality on dosimetry and the accuracy-efficiency trade-off of Monte Carlo (MC)-based scatter estimation and voxel dosimetry methods. Our unified framework consists of three stages: convolutional neural network (CNN)-based bremsstrahlung scatter estimation, SPECT reconstruction with scatter correction (SC) and absorbed dose-rate map generation with a residual learning network (DblurDoseNet). The input to the framework is the measured SPECT projections and CT, and the output is the absorbed dose-rate map. For training and testing under realistic conditions, we generated a series of virtual patient phantom activity/density maps from post-therapy images of patients treated with 90Y-RE at our clinic. To train the scatter estimation network, we use the scatter projections for phantoms generated from MC simulation as the ground truth (GT). To train the dosimetry network, we use MC dose-rate maps generated directly from the activity/density maps of phantoms as the GT (Phantom + MC Dose). We compared performance of our framework (SPECT w/CNN SC + DblurDoseNet) and MC dosimetry (SPECT w/CNN SC + MC Dose) using normalized root mean square error (NRMSE) and normalized mean absolute error (NMAE) relative to GT. When testing on virtual patient phantoms, our CNN predicted scatter projections had NRMSE of 4.0% ± 0.7% on average. For the SPECT reconstruction with CNN SC, we observed a significant improvement on NRMSE (9.2% ± 1.7%), compared to reconstructions with no SC (149.5% ± 31.2%). In terms of virtual patient dose-rate estimation, SPECT w/CNN SC + DblurDoseNet had a NMAE of 8.6% ± 5.7% and 5.4% ± 4.8% in lesions and healthy livers, respectively; compared to 24.0% ± 6.1% and 17.7% ± 2.1% for SPECT w/CNN SC + MC Dose. In patient dose-rate maps, though no GT was available, we observed sharper lesion boundaries and increased lesion-to-background ratios with our framework. For a typical patient data set, the trained networks took ~ 1 s to generate the scatter estimate and ~ 20 s to generate the dose-rate map (matrix size: 512 × 512 × 194) on a single GPU (NVIDIA V100). Our deep learning framework, trained using true activity/density maps, has the potential to outperform non-learning voxel dosimetry methods such as MC that are dependent on SPECT image quality. Across comprehensive testing and evaluations on multiple targeted lesions and healthy livers in virtual patients, our proposed deep learning framework demonstrated higher (66% on average in terms of NMAE) estimation accuracy than the current "gold-standard" MC method. The enhanced computing speed with our framework without sacrificing accuracy is highly relevant for clinical dosimetry following 90Y-RE.


Data Integrity-Based Methodology and Checklist for Identifying Implementation Risks of Physiological Sensing in Mobile Health Projects: Quantitative and Qualitative Analysis.
Mobile health (mHealth) technologies have the potential to bring health care closer to people with otherwise limited access to adequate health care. However, physiological monitoring using mobile medical sensors is not yet widely used as adding biomedical sensors to mHealth projects inherently introduces new challenges. Thus far, no methodology exists to systematically evaluate these implementation challenges and identify the related risks. This study aimed to facilitate the implementation of mHealth initiatives with mobile physiological sensing in constrained health systems by developing a methodology to systematically evaluate potential challenges and implementation risks. We performed a quantitative analysis of physiological data obtained from a randomized household intervention trial that implemented sensor-based mHealth tools (pulse oximetry combined with a respiratory rate assessment app) to monitor health outcomes of 317 children (aged 6-36 months) that were visited weekly by 1 of 9 field workers in a rural Peruvian setting. The analysis focused on data integrity such as data completeness and signal quality. In addition, we performed a qualitative analysis of pretrial usability and semistructured posttrial interviews with a subset of app users (7 field workers and 7 health care center staff members) focusing on data integrity and reasons for loss thereof. Common themes were identified using a content analysis approach. Risk factors of each theme were detailed and then generalized and expanded into a checklist by reviewing 8 mHealth projects from the literature. An expert panel evaluated the checklist during 2 iterations until agreement between the 5 experts was achieved. Pulse oximetry signals were recorded in 78.36% (12,098/15,439) of subject visits where tablets were used. Signal quality decreased for 1 and increased for 7 field workers over time (1 excluded). Usability issues were addressed and the workflow was improved. Users considered the app easy and logical to use. In the qualitative analysis, we constructed a thematic map with the causes of low data integrity. We sorted them into 5 main challenge categories: environment, technology, user skills, user motivation, and subject engagement. The obtained categories were translated into detailed risk factors and presented in the form of an actionable checklist to evaluate possible implementation risks. By visually inspecting the checklist, open issues and sources for potential risks can be easily identified. We developed a data integrity-based methodology to assess the potential challenges and risks of sensor-based mHealth projects. Aiming at improving data integrity, implementers can focus on the evaluation of environment, technology, user skills, user motivation, and subject engagement challenges. We provide a checklist to assist mHealth implementers with a structured evaluation protocol when planning and preparing projects.


Critical assessment of genome-scale metabolic models of Arabidopsis thaliana.
Genome-scale metabolic models (GEMs) have enabled researchers to perform systems-level studies of living organisms. Flux balance analysis (FBA), as a constraint-based technique, enables computation of reaction fluxes and prediction of the metabolic phenotypes of a cell under a set of specified conditions. The quality of a GEM is important for obtaining accurate predictions. In this study, we evaluated the quality of five available GEMs for Arabidopsis thaliana from various points of views. To do this, we inspected some of their important features, including the number of reactions with well-defined gene-protein-reaction rules, number of blocked reactions, mass-unbalanced reactions, prediction accuracy in the simulation of key metabolic functions and existence of erroneous energy generating cycles (EGCs). All of the models were found to include some mass-unbalanced reactions. Moreover, four out of five models were found to include EGCs. However, Aracell includes the maximum number of blocked reactions, which suggests the presence of several incomplete pathways. These results clearly show that simulation by using these models may result in erroneous predictions and all of the publicly available GEMs for A. thaliana require extensive curations before being applied in practice.


Predictive modelling of complex agronomic and biological systems.
Biological systems are tremendously complex in their functioning and regulation. Studying the multifaceted behaviour and describing the performance of such complexity has challenged the scientific community for years. The reduction of real-world intricacy into simple descriptive models has therefore convinced many researchers of the usefulness of introducing mathematics into biological sciences. Predictive modelling takes such an approach another step further in that it takes advantage of existing knowledge to project the performance of a system in alternating scenarios. The ever growing amounts of available data generated by assessing biological systems at increasingly higher detail provide unique opportunities for future modelling and experiment design. Here we aim to provide an overview of the progress made in modelling over time and the currently prevalent approaches for iterative modelling cycles in modern biology. We will further argue for the importance of versatility in modelling approaches, including parameter estimation, model reduction and network reconstruction. Finally, we will discuss the difficulties in overcoming the mathematical interpretation of in vivo complexity and address some of the future challenges lying ahead.


Kinetics of glucose transport by the perfused mid-gut of the freshwater prawn Macrobrachium rosenberg ii.
1. Mucosal influx of [3H]glucose was examined in the mid-gut of a freshwater prawn, Macrobrachium rosenbergii, using an in vitro perfusion technique. 2. [3H]glucose transfer across the apical cell membrane of the epithelium exhibited Michaelis-Menten kinetics (Jmax.in = 0-15 mumole glucose equiv/g. min, Kt = 0-17 mM). Under Na-free conditions, glucose influx was significantly reduced and a linear function of substrate concentration, indicative of either slow cellular diffusion (KD = 7-6 X 10(3) mumole glucose equiv/g. min. mM) or a facilitated process with a low carrier affinity for the sugar. 3. Phlorizin was a potent competitive inhibitor of glucose influx (K1 = 3-6 X10(-3) mM), galactose and 3-O-methylglucose (3-O-MG) were weak inhibitors, and fructose had no evident effect on glucose uptake. Azide, but not iodoacetate (IAA), significantly depressed influx. 4. Absorbed [3H]glucose was rapidly metabolized by the mid-gut. The majority of accumulated activity within the tissue was in the form of phosphorylated compounds and tritiated water (THO), while only 0-3% was recovered as a free-glucose. 5. Preliminary studies examining transmural [3-H]glucose transport, however, demonstrated a significant net mucosal to serosal free-glucose flux across the prawn mid-gut which was Na-dependent and IAA- and phlorizin-sensitive. Two alternative interpretations of the data are advanced as possible mechanisms for transepithelial glucose transport: (1) group translocation, or (2) the operation of an energized, high affinity, baso-lateral sugar transport carrier.


Imaging of a clinically relevant stroke model: glucose hypermetabolism revisited.
Ischemic stroke has been shown to cause hypermetabolism of glucose in the ischemic penumbra. Experimental and clinical data indicate that infarct-related systemic hyperglycemia is a potential therapeutic target in acute stroke. However, clinical studies aiming for glucose control in acute stroke have neither improved functional outcome nor reduced mortality. Thus, further studies on glucose metabolism in the ischemic brain are warranted. We used a rat model of stroke that preserves collateral flow. The animals were analyzed by [2-(18)F]-2-fluoro-2-deoxy-d-glucose positron emission tomography or magnetic resonance imaging during 90-minute occlusion of the middle cerebral artery and during 60 minutes after reperfusion. Results were correlated to magnetic resonance imaging of cerebral blood flow, diffusion of water, lactate formation, and histological data on cell death and blood-brain barrier breakdown. We detected an increased [2-(18)F]-2-fluoro-2-deoxy-d-glucose uptake within ischemic regions succumbing to infarction and in the peri-infarct region. Magnetic resonance imaging revealed impairment of blood flow to ischemic levels in the infarct and a reduction of cerebral blood flow in the peri-infarct region. Magnetic resonance spectroscopy revealed lactate in the ischemic region and absence of lactate in the peri-infarct region. Immunohistochemical analyses revealed apoptosis and blood-brain barrier breakdown within the infarct. The increased uptake of [2-(18)F]-2-fluoro-2-deoxy-d-glucose in cerebral ischemia most likely reflects hypermetabolism of glucose meeting increased energy needs of ischemic and hypoperfused brain tissue, and it occurs under both anaerobic and aerobic conditions measured by local lactate production. Infarct-related systemic hyperglycemia could serve to facilitate glucose supply to the ischemic brain. Glycemic control by insulin treatment could negatively influence this mechanism.


Using Machine Learning to Predict Young People's Internet Health and Social Service Information Seeking.
Machine learning creates new opportunities to design digital health interventions for youth at risk for acquiring HIV (YARH), capitalizing on YARH's health information seeking on the internet. To date, researchers have focused on descriptive analyses that associate individual factors with health-seeking behaviors, without estimating of the strength of these predictive models. We developed predictive models by applying machine learning methods (i.e., elastic net and lasso regression models) to YARH's self-reports of internet use. The YARH were aged 14-24 years old (N = 1287) from Los Angeles and New Orleans. Models were fit to three binary indicators of YARH's lifetime internet searches for general health, sexual and reproductive health (SRH), and social service information. YARH responses regarding internet health information seeking were fed into machine learning models with potential predictor variables based on findings from previous research, including sociodemographic characteristics, sexual and gender minority identity, healthcare access and engagement, sexual behavior, substance use, and mental health. About half of the YARH reported seeking general health and SRH information and 26% sought social service information. Areas under the ROC curve (≥ .75) indicated strong predictive models and results were consistent with the existing literature. For example, higher education and sexual minority identification was associated with seeking general health, SRH, and social service information. New findings also emerged. Cisgender identity versus transgender and non-binary identities was associated with lower odds of general health, SRH, and social service information seeking. Experiencing intimate partner violence was associated with higher odds of seeking general health, SRH, and social service information. Findings demonstrate the ability to develop predictive models to inform targeted health information dissemination strategies but underscore the need to better understand health disparities that can be operationalized as predictors in machine learning algorithms.


Cruise ship pathologies in remote regions.
Navigations on cruise ships are rising and tend to spread to remote areas like polar regions. Our aim was to assess the prevalence of pathologies encountered on a cruise ship navigating in remote areas including Polar Seas. A prospective observational, descriptive and cross-sectional study was conducted aboard a cruise ship with an overall capacity of 200-264 passengers and 140 crewmembers, sailing in remote areas as the Arctic Ocean and the Antarctic Peninsula over a period of 205 days. The database was built on all first consultations for passengers and crewmembers done by the onboard physician. Each symptom and diagnosis was coded according to the "International Classification of Primary Care, 2nd edition". For statistical analysis, the quantitative data were expressed as mean ± standard deviation and qualitative data as percentages. The percentages were compared using a c2 test corrected according to the Yates' method or by a Fisher test when appropriate. A total of 446 diagnoses were studied on the 910 consultations originally included (13.7% of the people on board). The median age for the passengers and the crewmembers was respectively 68 (age ranging from 12 to 90) years and 31 (18-62) years. Likewise, the sex ratio (male/female) was 0.98 and 3.23. Infectious diseases were predominant (prevalence of 43.7%). Among them, respiratory infections were the most common and gastroenteritis seemed to be more frequent in passengers (prevalence of 11.5% vs. 5%, p = 0.10). Cutaneous pathologies were more frequent in crewmembers (prevalence of 26.6% vs. 18.7%, p = 0.04) and allergic dermatitis was the second most frequent in this group of patients (prevalence of 7.2%). Cardiovascular diseases, more common in passengers (p = 0.05), represented 4% of all diagnoses. Two cases of phlebitis, one stroke and one subacute heart failure were diagnosed. Among traumatic injuries, cutaneous traumas were the commonest (prevalence of 76.5%). Musculoskeletal traumas were more common in passengers (p = 0.04). An acute gastrointestinal haemorrhage required a medical evacuation from the Antarctic Peninsula. The physician should be prepared to face emergency cases by developing personal expertise specific to maritime medicine in remote areas. Highlighting the particularity of cases handled in remote areas, our results should also pave the way of the development of medical protocols for ships lacking physician.


Amitriptyline- and mianserin-induced changes in acquisition of paired-association learning-task.
1 The double-blind study on twenty healthy students was an attempt at assessing the effects of 2-week's treatment with amitriptyline (25 mg three times a day) and mianserin (10 mg three times a day), each alone or separatively inbibed with alcohol (0.5 g/kg) on the immediate memory and on the acquisition of a paired-association learning-task. 2 Amitriptyline impaired both the short-term memory-span and acquisition, and alcohol potentiated these effects. The action of mianserin did not deviate significantly from that of the placebo, and it also failed to interact with alcohol. 3 It is concluded that the decrement in learning capacity, that occurs after the 2-week's treatment with therapeutic doses of amitriptyline, reflects changes in both the intrinsic and the regulatory mechanisms of learning.


Slow maturation of arterio-venous fistula in seven uremic patients: use of Ash Split Cath(R) as temporary, prolonged vascular access.
The temporary vascular access is the essential condition required to perfrom hemodialysis in uremic patients in the absence of a permanent and utilizable vascular access. The cannulation of a central vein with a dual lumen catheter may be useful when a few weeks are required for the maturation of AVF. Longer times for AVF maturation (such as in diabetic patients and in aged patients) impose the use of a tunnelled catheter such as Tesio Catheter or Hickman Catheter which lead to minor complications and more efficient dialysis treatment. The Ash Split Cath(R), a recently introduced chronic hemodialysis catheter, provides dialysis via a transcutaneous portion containing a 14 French cylindrical shaped catheter with D-shaped lumens and a dacron cuff. Due to the slow maturing of AVF, in our Department the Ash Split Cath has been used in 7 uremic patients (3 males and 4 females) who required hemodialysis. The cannulation of the internal jugular vein was performed by an ultrasound assisted technique and the correct catheter position was verified by standard chest X-rays. The average blood flowrates were 250 ml/min, and the mean KT/V calculated in all patients one month after the beginning of the dialytic therapy was 1.09 +/- 0.02. In six patients the catheter was utilized for at least 4 months, in one patient for 8 months. The devices were easily removed when the patient's AVF was functional and usable. We found that the use of the Ash Split Cath as a temporary, prolonged vascular access in uremic patients was op-timal allowing for flexibility in organizing the dialysis treatment schedule and in yielding a good performance in the initial dialysis therapy. Moreover, this device allows, in these patients, a satisfactory dialysis efficiency.


Exemplars in vaccine delivery protocol: a case-study-based identification and evaluation of critical factors in achieving high and sustained childhood immunisation coverage in selected low-income and lower-middle-income countries.
Increases in global childhood vaccine delivery have led to decreases in morbidity from vaccine-preventable diseases. However, these improvements in vaccination have been heterogeneous, with some countries demonstrating greater levels of change and sustainability. Understanding what these high-performing countries have done differently and how their decision-making processes will support targeted improvements in childhood vaccine delivery. We studied three countries-Nepal, Senegal, Zambia-with exemplary improvements in coverage between 2000 and 2018 as part of the Exemplars in Global Health Programme. We apply established implementation science frameworks to understand the 'how' and 'why' underlying improvements in vaccine delivery and coverage. Through mixed-methods research, we will identify drivers of catalytic change in vaccine coverage and the decision-making process supporting these interventions and activities. Methods include quantitative analysis of available datasets and in-depth interviews and focus groups with key stakeholders in the global, national and subnational government and non-governmental organisation space, as well as community members and local health delivery system personnel. Working as a multinational and multidisciplinary team, and under oversight from all partner and national-level (where applicable) institutional review boards, we collect data from participants who provided informed consent. Findings are disseminated through a variety of forms, including peer-reviewed manuscripts related to country-specific case studies and vaccine system domain-specific analyses, presentations to key stakeholders in the global vaccine delivery space and narrative dissemination on the Exemplars.Health website.


Bone Metastasis of Prostate Cancer Can Be Therapeutically Targeted at the TBX2-WNT Signaling Axis.
Identification of factors that mediate visceral and bone metastatic spread and subsequent bone remodeling events is highly relevant to successful therapeutic intervention in advanced human prostate cancer. TBX2, a T-box family transcription factor that negatively regulates cell-cycle inhibitor p21, plays critical roles during embryonic development, and recent studies have highlighted its role in cancer. Here, we report that TBX2 is overexpressed in human prostate cancer specimens and bone metastases from xenograft mouse models of human prostate cancer. Blocking endogenous TBX2 expression in PC3 and ARCaPM prostate cancer cell models using a dominant-negative construct resulted in decreased tumor cell proliferation, colony formation, and invasion in vitro Blocking endogenous TBX2 in human prostate cancer mouse xenografts decreased invasion and abrogation of bone and soft tissue metastasis. Furthermore, blocking endogenous TBX2 in prostate cancer cells dramatically reduced bone-colonizing capability through reduced tumor cell growth and bone remodeling in an intratibial mouse model. TBX2 acted in trans by promoting transcription of the canonical WNT (WNT3A) promoter. Genetically rescuing WNT3A levels in prostate cancer cells with endogenously blocked TBX2 partially restored the TBX2-induced prostate cancer metastatic capability in mice. Conversely, WNT3A-neutralizing antibodies or WNT antagonist SFRP-2 blocked TBX2-induced invasion. Our findings highlight TBX2 as a novel therapeutic target upstream of WNT3A, where WNT3A antagonists could be novel agents for the treatment of metastasis and for skeletal complications in prostate cancer patients. Cancer Res; 77(6); 1331-44. ©2017 AACR.


Reachability analysis of real-time systems using time Petri nets.
Time Petri nets (TPNs) are a popular Petri net model for specification and verification of real-time systems. A fundamental and most widely applied method for analyzing Petri nets is reachability analysis. The existing technique for reachability analysis of TPNs, however, is not suitable for timing property verification because one cannot derive end-to-end delay in task execution, an important issue for time-critical systems, from the reachability tree constructed using the technique. In this paper, we present a new reachability based analysis technique for TPNs for timing property analysis and verification that effectively addresses the problem. Our technique is based on a concept called clock-stamped state class (CS-class). With the reachability tree generated based on CS-classes, we can directly compute the end-to-end time delay in task execution. Moreover, a CS-class can be uniquely mapped to a traditional state class based on which the conventional reachability tree is constructed. Therefore, our CS-class-based analysis technique is more general than the existing technique. We show how to apply this technique to timing property verification of the TPN model of a command and control (C2) system.


Intact tactile anisotropy despite altered hand perception in complex regional pain syndrome: rethinking the role of the primary sensory cortex in tactile and perceptual dysfunction.
Complex Regional Pain Syndrome (CRPS) is characterised by pain, autonomic, sensory and motor abnormalities. It is associated with changes in the primary somatosensory cortex (S1 representation), reductions in tactile sensitivity (tested by two-point discrimination), and alterations in perceived hand size or shape (hand perception). The frequent co-occurrence of these three phenomena has led to the assumption that S1 changes underlie tactile sensitivity and perceptual disturbances. However, studies underpinning such a presumed relationship use tactile sensitivity paradigms that involve the processing of both non-spatial and spatial cues. Here, we used a task that evaluates anisotropy (i.e., orientation-dependency; a feature of peripheral and S1 representation) to interrogate spatial processing of tactile input in CRPS and its relation to hand perception. People with upper limb CRPS (n = 14) and controls with (n = 15) or without pain (n = 19) judged tactile distances between stimuli-pairs applied across and along the back of either hand to provide measures of tactile anisotropy. Hand perception was evaluated using a visual scaling task and questionnaires. Data were analysed with generalised estimating equations. Contrary to our hypotheses, tactile anisotropy was bilaterally preserved in CRPS, and the magnitude of anisotropic perception bias was comparable between groups. Hand perception was distorted in CRPS but not related to the magnitude of anisotropy or bias. Our results suggest against impairments in spatial processing of tactile input, and by implication S1 representation, as the cause of distorted hand perception in CRPS. Further work is warranted to elucidate the mechanisms of somatosensory dysfunction and distorted hand perception in CRPS.


The envelope-associated 22K protein of human respiratory syncytial virus: nucleotide sequence of the mRNA and a related polytranscript.
We recently determined that respiratory syncytial virus (strain A2) encodes a fourth unique envelope-associated virion protein that has molecular weight of approximately 24,000, as estimated by gel electrophoresis. The nucleotide sequence of the mRNA encoding this novel protein has now been determined from five cDNA clones, including three that contain the complete mRNA sequence. The complete mRNA sequence is 957 nucleotides, exclusive of polyadenylate, and contains two partially overlapping open reading frames. The 5'-proximal open reading frame is favored for utilization by the criteria of the location and sequence of its translational start site. Furthermore, the calculated molecular weight of the encoded protein, 22,153, is in agreement with the previous estimate of 24,000 for the authentic protein identified by hybrid selection and in vitro translation. The sequence of the predicted protein, now designated the 22K protein, contains 194 amino acids, is relatively hydrophilic, and appears to be the most basic of the respiratory syncytial virus proteins. The mRNA also contains a second, internal open reading frame which would encode a protein of 90 amino acids. However, no evidence for this translation product is known. The first nine nucleotides in the mRNA sequence, 5'-GGGGCAAAU, are identical to the conserved sequence identified previously at the 5' termini of seven other respiratory syncytial viral mRNAs; the sequence at the 3' end of the 22K mRNA, 5'. . . AGUUAUUU-polyadenylate, contains the elements of the previously identified 3'-terminal consensus sequence for respiratory syncytial virus mRNAs, AGUUAA(N)1-4-polyadenylate (P. L. Collins, Y. T. Huang, and G. W. Wertz, Proc. Natl. Acad. Sci. U.S.A. 81:7683-7687). In addition, we present and describe the intergenic sequence of a dicistronic RNA derived from readthrough of the F and 22K protein genes.


[Burch-Schneider Ring in Revision Surgery of THR.].
The authors evaluate the results of 16 revision surgeries of the hip joint with the application of Burch-Schneider ring in 3A and 3B Paprosky type of acetabular defect on average 4 years after the primary surgery. According to the subjective evaluation 9 of them were very good, 6 of them were good and 1 result was satisfactory. The average Harris Hip Score was 88 points. They did not encounter any evident or probable loosening of the cup. In all patients bone allografts were fully integrated. The new bone created under the Burch-Schneider ring can be in future used for the revision surgery, if need be. Burch-Schneider ring can be successfully used for the treatment of significant defects of acetabulum in revision surgery of the hip joint. Key words: THR, revision surgery.


Intestinal permeability tests in coeliac disease.
Intestinal permeability tests have been used to screen for a wide range of small intestinal diseases, including coeliac disease and enteric infections. Several probe molecules have been used to investigate intestinal permeability including monosaccharides, disaccharides, 51Cr-EDTA and polyethyleneglycol. While many factors may affect intestinal permeability tests, the use of two probe molecules, for example, lactulose and mannitol, and the expression of the result as a ratio minimises the effects of these extraneous factors. Rendering the test solution hyperosmolar was also found to increase the sensitivity of the test in detecting coeliac disease. Intestinal permeability is characteristically elevated in untreated coeliac disease, with a sensitivity of up to 96% for the dual sugar techniques. The reason for this is a consistent increase in the absorption of lactulose (via the paracellular route) due to increased "leakiness" of the intestine and a reduction in the absorption of mannitol (via the transcellular route) due to a reduction in surface area as a result of villous atrophy. The intestinal permeability test allows subjects to be selected for jejunal biopsy in whom the clinical features are compatible with coeliac disease and in timing a follow-up biopsy. It has been postulated that raised intestinal permeability may be involved in the pathogenesis of coeliac disease. Recently, serum measurements of the probe molecules may have a valuable role, particularly in paediatric patients. Sucrose permeability has also been proposed as an accurate marker of adult coeliac disease and shows promise as a noninvasive test.


Indolethylamine-N-methyltransferase activity in psychiatric patients and controls.
The level of INMT activity was determined in the sera of 29 psychiatric patients and 11 healthy controls from St. Louis; and in 13 psychiatric patients and 15 healthy controls from Chicago. The level of enzyme activity in the serum of paranoid schizophrenics in the St. Louis group was significantly higher than in other types of schizophrenics. The mean level of enzyme activity in the serum in nonschizophrenic psychiatric patients in the Chicago group was significantly higher than that in the same group of patients from St. Louis. The serum level of INMT activity in all psychiatric patients and schizophrenic patients from St. Louis was positively correlated with severity of delusions. The only significant difference in the Chicago patients was that the occurrence of depressive features was greater in the group of patients with a low serum INMT level than in the group with a high enzyme level.


Longitudinal changes in infant body composition: association with childhood obesity.
Rapid weight gain in infancy has been established as a risk factor for the development of later obesity. We aimed to investigate the role of changes in infant body composition (assessed via total body electrical conductivity) on the development of overweight/obesity in mid-childhood. Fifty-three term infants were evaluated at birth, three times during infancy and in mid-childhood. Logistic regression was used to determine associations between rates of total weight gain, fat mass gain and lean mass gain during infancy and later overweight/obesity (defined as body mass index [BMI] ≥85th percentile), adjusted for birth weight and parent education. At follow-up (age 9.0 ± 1.8 years), 30% were overweight/obese. More rapid total weight gain from 0 to 4 months was associated with twofold odds (odds ratio [OR] 1.98, 95% confidence interval [CI] 1.05-3.74, P = 0.04) of overweight/obesity in mid-childhood. From 0 to 8 months, more rapid weight gain was associated with nearly fivefold odds (OR 4.76, 95% CI 1.05-21.5, P = 0.04), and more rapid fat mass gain was associated with eightfold odds (OR 8.03, 95% CI 1.11-58.2, P = 0.04) of later overweight/obesity. This exploratory study suggests that rapid weight gain, especially fat mass gain, in earlier infancy predisposes to mid-childhood overweight/obesity.


Immunologic factors in primary progressive multiple sclerosis.
Primary progressive multiple sclerosis (PPMS) is clinically characterized by progression without remission or relapse, in contrast to relapsing forms of MS. Pathologic and imaging findings also indicate that PPMS differs from relapsing forms. Recent studies examining potential immunologic differences among MS forms suggest that cytokine and adhesion molecule expression profiles, but not chemokine receptor profiles, in PPMS patients resemble those in healthy controls more than those in patients with relapsing MS. However, the significance of these findings remains to be fully elucidated, and there is little additional evidence as yet of marked differences among MS forms based on immunologic characteristics. Of interest are the recent demonstrations that activated immune cells produce brain-derived neurotrophic factor (BDNF), a neuroprotective factor, in MS lesions, that BDNF receptors are located in MS tissue, and that glatiramer acetate (GA)-specific T cells produce BDNF irrespective of T helper cell Th1 or Th2 phenotype. These findings both support the concept of a protective inflammation in MS and suggest an additional mechanism of action for the clinical effects of GA therapy in MS. Whether and to what extent this mechanism is present in different MS forms remain to be clarified.


Analysis of EPA and DHA in the viscera of marine fish using gas chromatography.
The viscera of 10 kinds of marine fishes were collected for fish oil extraction and detection of DHA and EPA, two most important polyunsaturated fatty acids. The fish oil extraction ratio for the evaluated fishes varied from 0.95% to 10.18% (wt%). Pseudosciaena crocea presented the highest fish oil yield, followed by Mustelus manazo, Hippoglossus and Sciaenopsocellatus. A gas chromatography method was then established for analysis of EPA/DHA. The EPA concentration (in methyl ester form) in the fish oil varied from 1.39 to 10.65(mg/g). Epinephelus awoara presented the highest EPA concentration (p<0.05), followed by Epinephelussp, Sciaenopsocellatus and Hippoglossus. The DHA concentration (in methyl ester form) in the fish oil varied from 0.58 to 37.02 (mg/g). Epinephelus awoara presented the highest DHA concentration (p<0.05), followed by Sciaenopsocellatus, Pseudosciaena crocea and Hippoglossus. No strict positive correlation between the EPA/DHA concentration and the sea depth where the fish live was observed. The fishes living in middle depth presented highest EPA/DHA concentration.


Utility of the sonographic whirlpool sign in diagnosing midgut volvulus in patients with atypical clinical presentations.
To illustrate cases of midgut volvulus in 7 children, of whom only 2 were neonates, diagnosed initially by sonography using the whirlpool sign, and to emphasize the importance of considering malrotation and midgut volvulus in the differential diagnosis of acute or chronic abdominal pain in those beyond the neonatal period, the usual time of presentation for midgut volvulus. The sonography computer database of a large pediatric hospital was reviewed over a 26-month period for sonographic reports that contained the key words midgut volvulus and whirlpool sign. From November 1999 until January 2002, 7 patients had a sonographic diagnosis of midgut volvulus on the basis of the whirlpool sign. Sonographic and surgical records of these patients were reviewed retrospectively. In addition, the department computer database of all contrast-enhanced upper gastrointestinal examinations performed over the same period was searched using the key words midgut volvulus, and an additional 2 cases of midgut volvulus were found, neither of which had undergone a prior or subsequent sonographic study. Seven patients, ranging in age from 1 day to 5 1/2 years were found to have the whirlpool sign on sonography performed for acute abdominal symptoms, including projectile nonbilious emesis, colicky abdominal pain with suspicion of intussusception, palpable abdominal masses, right lower quadrant abdominal pain with suspected appendicitis, and chronic diarrhea. Six of the 7 cases of midgut volvulus were confirmed with upper gastrointestinal examinations. One patient was taken directly to surgery on the basis of the sonographic findings alone. One patient had subsequent computed tomography of the abdomen for continued suspicion of an abdominal mass. All 7 cases were confirmed as midgut volvulus at surgery. The sonographic whirlpool sign is a valid and highly sensitive sign for the diagnosis of midgut volvulus secondary to malrotation, and sonographers should be aware that it frequently occurs beyond the neonatal period with symptoms other than bilious emesis.


A generalization of the state-specific complete-active-space coupled-cluster method for calculating electronic excited states.
The complete-active-space coupled-cluster approach with single and double excitations (CASCCSD) based on the ansatz of Oliphant and Adamowicz [J. Chem. Phys. 94, 1229 (1991); 96, 3739 (1992)] is used to derive an approach termed XCASCCSD for calculating potential energy surfaces of ground and excited electronic states with different multiplicities and symmetries. The XCASCCSD approach explicitly includes a procedure for spin and spatial orbital-momentum symmetry adaptation of the wave function that has allowed us to consider states with degenerate formal references. The XCASCCSD method is applied to calculate potential energy surfaces of the ground and some lowest singlet and triplet excited states of the FH and C(2) molecules. Some states of C(2) are known to have a very strong "multireference" character making their description difficult with single-reference methods. The problem of the change of the formal reference determinant along the potential energy surface is discussed. Also, vertical excitation energies of formaldehyde calculated with the XCASCCSD approach are presented.


Pilot study on the clinical and microbiological effect of subgingival glycine powder air polishing using a cannula-like jet.
To assess the efficacy of subgingival glycine powder air polishing (GPAP) during supportive periodontal therapy (SPT). Each quadrant of 25 subjects was randomly assigned to the following treatments: subgingival scaling with hand instruments (SRP), GPAP, subgingival ultrasonic debridement (UD) and no subgingival treatment (NT). Clinical recordings included the following: probing pocket depth (PPD), gingival recession (RE), clinical attachment level (CAL), Gingival and Plaque Index. Subgingival plaque samples were taken from two sites >4 mm per quadrant. Therapy, recordings and microbial sampling were performed at baseline, 3 and 6 months, while at 1 month only clinical recordings and sampling were performed. Subgingival samples were analysed using 'checkerboard' DNA-DNA hybridization for Porphyromonas gingivalis, Tannerrella forsythia and Treponema denticola. All groups were homogeneous at baseline for the clinical parameters assessed. The GPAP group displayed statistically significant higher PPD compared to SRP and UD at 1, 3 and 6 months and no statistical differences with the 'no treatment' group at all time points. At 1 month, the GPAP group displayed statistically significantly higher levels of CAL compared to SRP, while at 3 and 6 months statistically significant differences were observed with groups assigned to SRP and UD. No differences were observed among groups for RE, PI, GI and numbers of the investigated bacteria at any time point. On the basis of clinical and microbiological data, this study does not support the superiority of GPAP as sole treatment over SRP or subgingival ultrasonic scaling.


Psychedelic Communitas: Intersubjective Experience During Psychedelic Group Sessions Predicts Enduring Changes in Psychological Wellbeing and Social Connectedness.
Background: Recent years have seen a resurgence of research on the potential of psychedelic substances to treat addictive and mood disorders. Historically and contemporarily, psychedelic studies have emphasized the importance of contextual elements ('set and setting') in modulating acute drug effects, and ultimately, influencing long-term outcomes. Nevertheless, current small-scale clinical and laboratory studies have tended to bypass a ubiquitous contextual feature of naturalistic psychedelic use: its social dimension. This study introduces and psychometrically validates an adapted Communitas Scale, assessing acute relational experiences of perceived togetherness and shared humanity, in order to investigate psychosocial mechanisms pertinent to psychedelic ceremonies and retreats. Methods: In this observational, web-based survey study, participants (N = 886) were measured across five successive time-points: 2 weeks before, hours before, and the day after a psychedelic ceremony; as well as the day after, and 4 weeks after leaving the ceremony location. Demographics, psychological traits and state variables were assessed pre-ceremony, in addition to changes in psychological wellbeing and social connectedness from before to after the retreat, as primary outcomes. Using correlational and multiple regression (path) analyses, predictive relationships between psychosocial 'set and setting' variables, communitas, and long-term outcomes were explored. Results: The adapted Communitas Scale demonstrated substantial internal consistency (Cronbach's alpha = 0.92) and construct validity in comparison with validated measures of intra-subjective (visual, mystical, challenging experiences questionnaires) and inter-subjective (perceived emotional synchrony, identity fusion) experiences. Furthermore, communitas during ceremony was significantly correlated with increases in psychological wellbeing (r = 0.22), social connectedness (r = 0.25), and other salient mental health outcomes. Path analyses revealed that the effect of ceremony-communitas on long-term outcomes was fully mediated by communitas experienced in reference to the retreat overall, and that the extent of personal sharing or 'self-disclosure' contributed to this process. A positive relationship between participants and facilitators, and the perceived impact of emotional support, facilitated the emergence of communitas. Conclusion: Highlighting the importance of intersubjective experience, rapport, and emotional support for long-term outcomes of psychedelic use, this first quantitative examination of psychosocial factors in guided psychedelic settings is a significant step toward evidence-based benefit-maximization guidelines for collective psychedelic use.


Disseminated tetracycline resistance in oral streptococci: implication of a conjugative transposon.
A DNA sequence specifying tetracycline resistance (Tcr) has been previously cloned from a clinical isolate of Streptococcus mutans designated U202 (J. A. Tobian and F. L. Macrina, J. Bacteriol. 152:215-222, 1982). We used this sequence as a molecular probe in studying the dissemination of Tcr among oral streptococcal species isolated from patients treated with tetracycline. Eleven strains (including S. sanguis I, S. sanguis II, S. mitis, and S. salivarius) from seven patients were examined by Southern blot analysis. Seven strains showed strong hybridization to the Tcr probe, two showed weak hybridization, and two did not display detectable hybridization. Based on previous characterization of the cloned sequence, our data suggest the dissemination of the tetM class of resistance determinants among these oral streptococci. One of the clinical S. sanguis I isolates studied was able to transfer its Tcr phenotype to other oral streptococci and to enteric streptococci in the absence of plasmid DNA. This transfer appeared to be conjugation-like on the basis of its insensitivity to DNase and its dependence on intimate cell-to-cell contact. Using the cloned Tcr sequence, we were able to study the progeny of the matings. Our data suggest that this resistance transfer element occupies a chromosomal location in streptococcal cells and that it strongly resembles the conjugative transposon Tn916 in its behavior.


Mechanical pump dacryoreservoirs.
The two most important characteristics of the natural tear are its chemophysical properties and the continuity of its delivery. The chemical characteristics of tears are very difficult to reproduce since they contain more than 200 different components. In severe dry eyes - due to their relative short residence - manual application of artificial tears from small bottles to the eye has to be performed every 5-10 min, while at night no lubrication at all is provided. Previously, for continuous lubrication of the ocular surface, dacryoreservoirs attached to spectacles or placed in pockets of the patient's clothes were used, but were often complicated by infection. A new option is the implantable pump dacryoreservoir. In 21 patients with a Schirmer test without anaesthesia of <2mm in 5 min, a Medtronic 60-ml reservoir was implanted into a pocket under the subcutaneous tissues of the anterior abdominal wall and connected to a silicone catheter that ascended subcutaneously along the chest, neck and temple to the upper conjunctival fornix. The results of this procedure and complications associated with the implantation of reservoirs in general - as evaluated on the basis of a Medline search - are presented. Postoperatively all patients reported a dramatic improvement of dry eye symptoms. Slit-lamp microscopy revealed a substantially prolonged break-up time and a reduction of signs of ocular surface disease such as superficial punctuate keratopathy and conjunctival hyperaemia. A penetrating keratoplasty was successfully performed in 2 eyes with an implanted dacryoreservoir and remained clear throughout the follow-up of up to 3.5 years. No infections of the catheter reservoir were observed. Prominent parts of the catheter induced skin ulceration, but no infection, in 2 patients which was managed successfully by removal of the catheter, systemic antibiotics and subsequent re-implantation of a new catheter. A literature review showed that infection is not a frequent problem with implantable and external reservoirs. At present, this is the only safe and effective method able to maintain a continuous lubrication of the ocular surface with artificial tears, and the only one that allows corneal, conjunctival or limbal transplantations in severe dry eyes.


Application of an Integrated and Open Source Workflow for LC-HRMS Plant Metabolomics Studies. Case-Control Study: Metabolic Changes of Maize in Response to Fusarium verticillioides Infection.
Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) represents the most powerful metabolomics platform to investigate biological systems. Reproducible and standardized workflows allow obtaining a meaningful biological interpretation. The purpose of this study was to set up and apply an open-source workflow for LC-HRMS plant metabolomics studies. Key steps of the proposed workflow were as follows: (1) experimental design, (2) sample preparation, (3) LC-HRMS analysis, (4) data processing, (5) custom database search, (6) statistical analysis, (7) compound identification, and (8) biochemical interpretation. Its applicability was evaluated through the study of metabolomics changes of two maize recombinant inbred lines with contrasting phenotypes with respect to disease severity after Fusarium verticillioides infection of seedlings. Analysis of data from the case-control study revealed abundance change in metabolites belonging to different metabolic pathways, including two amino acids (L-tryptophan and tyrosine), five flavonoids, and three N-hydroxynnamic acid amides.


Fever-range hyperthermia dynamically regulates lymphocyte delivery to high endothelial venules.
Fever is associated with increased survival during acute infection, although its mechanism of action is largely unknown. This study found evidence of an unexpectedly integrated mechanism by which fever-range temperatures stimulate lymphocyte homing to secondary lymphoid tissues by increasing L-selectin and alpha4beta7 integrin-dependent adhesive interactions between circulating lymphocytes and specialized high endothelial venules (HEV). Exposure of splenic lymphocytes in vivo to fever-like whole-body hyperthermia (WBH; 39.8 +/- 0.2 degrees C for 6 hours) stimulated both L-selectin and alpha4beta7 integrin-dependent adhesion of lymphocytes to HEV under shear conditions in lymph nodes and Peyer patches. The adhesiveness of HEV ligands for L-selectin and alpha4beta7 integrin (ie, peripheral lymph node addressin and mucosal addressin cell adhesion molecule-1) also increased during WBH or febrile responses associated with lipopolysaccharide-induced or turpentine-induced inflammation. Similar increases in HEV adhesion occurred during hyperthermia treatment of lymph node and Peyer patch organ cultures in vitro, indicating that the local lymphoid tissue microenvironment is sufficient for the hyperthermia response. In contrast, WBH did not augment adhesion in squamous endothelium of nonlymphoid tissues. Analysis of homing of alpha4beta7(hi) L-selectin(lo) murine TK1 cells and L-selectin(hi) alpha4beta7 integrin-negative 300.19/L-selectin transfectant cells showed that fever-range temperatures caused a 3- to 4-fold increase in L-selectin and alpha4beta7 integrin-dependent trafficking to secondary lymphoid tissues. Thus, enhanced lymphocyte delivery to HEV by febrile temperatures through bimodal regulation of lymphocyte and endothelial adhesion provides a novel mechanism to promote immune surveillance.


Voltammetric and pharmacological characterization of dopamine release from single exocytotic events at rat pheochromocytoma (PC12) cells.
Although rat pheochromocytoma (PC12) neurotransmitter storage vesicles are known to contain a variety of neurotransmitters including catecholamines, there is little evidence that the molecular species detected during amperometric monitoring of exocytosis is a catecholamine. Rather, as these are catecholamine-containing cells, one assumes catecholamines are released. Additionally, although the total amount of transmitter released can be quantified, it has been extremely difficult to evaluate the concentration at the point of release for each exocytosis event. Interpreting voltammograms obtained in the attoliter volume affected between the electrode and the cell and defined by the size of the exocytosis pore during exocytosis is an extreme analytical challenge. Here we use voltammetry of approximately 10(-19) mol released from individual exocytosis events to identify, along with pharmacological evidence, the released compound at PC12 cells as a catecholamine, most likely dopamine. The area of the electrode at which oxidation occurs following an exocytosis event is proportional to the temporal delay prior to acquisition of a voltammogram. This model allows determination of relative concentrations from individual release events and has been used to examine events at control cells and cells incubated with the dopamine precursor, L-3,4-dihydroxyphenylalanine (L-DOPA). Exposure to L-DOPA (100 microM for 1 h) results in 145 detectable events for 11 cells compared to 77 events for 29 control cells, clearly indicating that vesicles can be "loaded" with dopamine. However, the concentrations measured at the electrode surface provide similar distributions for both L-DOPA-treated and control cells. Cyclic voltammetric measurements of relative concentration for zeptomole levels of transmitter in attoliter volumes provide evidence that loading vesicles by increased transmitter synthesis does not lead to elevated concentrations at individual release sites.