Provider Perspectives on Principle-Adherent Practice in Empirically Supported Interventions for Emerging Adults with Serious Mental Health Conditions.
In recognition of the need to create new treatment approaches that will be appealing to and effective for emerging adults with serious mental health conditions, researchers have begun to create and evaluate programs and interventions that are specifically tailored to reflect the preferences and needs of the population. The literature that describes these new approaches-including both descriptions of interventions and guidelines based on expert consensus-expresses a high degree of agreement regarding practice principles that should guide intervention. However, beyond naming these principles, the literature provides little information about what the principles mean, or how principle-adherent practice can be recognized. This article describes a qualitative investigation of providers' understanding of principle-driven practice in the context of programs and interventions for emerging adults with serious mental health conditions. The goal was to learn about how providers conceptualize the principles that drive their practice, and how they describe principle-adherent practice.


Access to spine care for the poor and near poor.
Access to care for poor/near poor patients is a concerning and growing problem within the American system of medical care. The objective of this study was to examine the relationship between health insurance status and access to spine care among patients below 65 years of age eventually receiving treatment at our tertiary academic medical center. Descriptive study based on chart review and telephone interviews. Two groups of 64 patients each with surgical pathology of limited complexity and limited comorbidities, one with Medicaid insurance and one with private, commercial insurance. Reasons for referral, travel distance, travel time, frequency of visits, and proximity of fellowship-trained spinal surgeons. Two groups, each with 64 consecutive spine surgical patients, were studied and compared. Group One had "Medicaid" coverage and Group Two was privately insured. All patients (both groups) were treated surgically for similar pathology of limited complexity and had limited comorbidities. They were assessed to determine the difficulties they encountered in receiving care before referral to our medical center including factors such as referral by a local provider based on insurance status alone and travel time/distance/frequency to eventually receive care at our center. The availability of local care for these patients (fellowship-trained spine surgeons in their local area) was also assessed. The great majority (78%) of poor/near poor patients with Medicaid coverage from counties at some distance from (and local to) our center were referred/deferred on the basis of insurance status alone given surgical problems which could have comfortably been addressed by orthopedic surgeons, neurosurgeons, or fellowship-trained spine surgeons local to the patient. This difficulty in access to care results in a significant burden (measured in time/travel/costs) for these patients. The poor/near poor with Medicaid insurance have less access to local spine care than those with private, commercial health insurance. The implications (from both surgeon and patient perspectives) of this dilemma are discussed.


Imidazole-amino acids. Conformational switch under tautomer and pH change.
Replacement of the main chain peptide bond by imidazole ring seems to be a promising tool for the peptide-based drug design, due to the specific prototropic tautomeric as well as amphoteric properties. In this study, we present that both tautomer and pH change can cause a conformational switch of the studied residues of alanine (1-4) and dehydroalanine (5-8) with the C-terminal peptide group replaced by imidazole. The DFT methods are applied and an environment of increasing polarity is simulated. The conformational maps (Ramachandram diagrams) are presented and the stability of possible conformations is discussed. The neutral forms, tautomers τ (1) and π (2), adapt the conformations αRτ (φ, ψ = - 75°, - 114°) and C7eq (φ, ψ = - 75°, 66°), respectively. Their torsion angles ψ differ by about 180°, which results in a considerable impact on the peptide chain conformation. The cation form (3) adapts both these conformations, whereas the anion analogue (4) prefers the conformations C5 (φ, ψ = - 165°, - 178°) and β2 (φ, ψ ~ - 165°, - 3°). Dehydroamino acid analogues, the tautomers τ (5) and π (6) as well as the anion form (8), have a strong tendency toward the conformations β2 (φ, ψ = - 179°, 0°) and C5 (φ, ψ = - 180°, 180°). The preferences of the protonated imidazolium form (7) depend on the environment. The imidazole ring, acting as a donor or acceptor of the hydrogen bonds created within the studied residues, has a profound effect on the type of conformation.


Contribution of BRCA1 and BRCA2 to familial ovarian cancer: a gynecologic oncology group study.
The aim of the study was to determine the prevalence of BRCA1 and BRCA2 germline mutations among ovarian cancer patients ascertained to have a family history of ovarian cancer. Ovarian cancer patients were eligible if they had a family history of cancer that met any one of the following criteria: (1) a first-degree relative with ovarian cancer; (2) a second-degree relative with ovarian cancer plus a first-degree relative with breast cancer (diagnosed younger than 50 years of age); or (3) a first- and a second-degree relative with breast cancer (diagnosed younger than 50 years of age). The entire coding sequence of BRCA1 and exon 11 of BRCA2 were screened for germline alterations by single-strand conformation polymorphism analysis. Of 26 eligible patients screened for mutations, 12 had deleterious alterations, 8 in BRCA1 and 4 in BRCA2. A correlation was noted between the presence of a BRCA1 mutation and the strength of family history of breast ovarian cancer, with the likelihood of a mutation increasing with the number of affected relatives (P = 0.0002). No association was detected between the location of mutations in BRCA1 and the ratio of ovarian cancer cases relative to breast cancer (P = 0.28). Mutations in BRCA1 or BRCA2 are present in about 50% of ovarian cancer patients with at least one first-degree relative with disease, and in 70% of patients with two or more relatives with ovarian cancer.


The efficacy of combined estrogen and buspirone treatment in olivopontocerebellar atrophy.
Olivopontocerebellar atrophy (OPCA) is a chronic neurodegenerative disease with symptoms of cerebellar ataxia, parkinsonism, autonomic disturbances and ophthalmoplegia. Buspirone, a 5-HT1(A) agonist could constitute a symptomatic improvement in cerebellar dysfunction whereas estrogen has been investigated for neuroprotection. We conducted an open-labeled pilot trial to assess the efficacy of estrogen with buspirone treatment. Eighteen patients (7 male and 11 female) with OPCA were randomized into the buspirone (15 mg/day, n=9), or the combined treatment group (estrogen, 0.625 mg/d plus buspirone, n=9). For the clinical rating, International Cooperative Ataxia Rating Scale (ICARS) was used and dysarthria, gaze evoked nystagmus, finger to nose, pronation-supination alternating movement, knee-tibia test, and gait speed were evaluated for 12 months. Buspirone-treated group showed improvements in finger to nose and pronation-supination alternating movement test (p=0.046 and p=0.025, respectively). The combination group (Estrogen+buspirone), however, showed no improvement in cerebellar sub-scales compared to the baseline. Buspirone treatment showed feasible efficacies for OPCA, while the combined treatment of estrogen and buspirone failed to improve, suggesting estrogen may not have further benefit in cerebellar dysfunction.


Novel methodology to determine leakage power in standard cell library design.
In the most advanced technology nodes, leakage has become a major concern for integrated circuits designers. In addition, the leakage calculation using SPICE simulations takes a large amount of time for the entire library of standard cells for the different PVT (Process, Voltage, and Temperature) conditions. However IC Designers usually need a fast access to the leakage current estimation in order to validate their designs. In this context, the present paper proposes a new approach of the leakage estimation. The aim of this methodology is to predict the leakage of a standard cell accurately and in a short amount of time. This method is applied to all the cells of the used standard cell library and is based on mathematical models developed from the leakage physical equation. The results of the leakage calculated with this technique shows an average error of 5%. The context of this work is part of a fast generation of a full liberty file of a standard cell library using the curve fitting method.


Medical and social factors associated with the admission and discharge of acutely ill children.
To examine medical and sociodemographic factors involved in acute paediatric admission. To compare outcome of admission with factors present at time of admission. Prospective questionnaire based study of 887 consecutive emergency general paediatric admissions to five Yorkshire hospitals during two separate three week periods in summer and winter. Discharge diagnosis, length of stay. Most admissions (53%) occurred "out of hours" with a peak during the evening. Two thirds (64%) of patients were under 3 years of age and clinical problems varied with age. Self referral via an accident and emergency department occurred in one third and was more likely after a fit in older children and in more socioeconomically deprived children. The most frequent presenting problems were breathing difficulty (24%), fit (16%), and feverish illness (15%). One quarter (24%) were discharged within 24 hours and 61% spent, at most, one night in hospital. Length of stay was shorter for night admissions and longer for children with a discharge diagnosis of asthma. Although most children had mild, self limiting illnesses, serious illness was subsequently found in 13% and could not be predicted from the presenting problems. Current demand on emergency paediatric admission is mainly from young children with mild self limiting illnesses who spend one night or less in hospital. Changes in delivery of care to acutely ill children must take account of the pattern and nature of presenting problems and be rigorously audited to ensure that improvements in the health of children continue.


Synthesis and capillary electrophoretic analysis of enantiomerically enriched reference standards of MDMA and its main metabolites.
Enantiomerically-enriched (S)-3,4-methylenedioxymethamphetamine (MDMA) and its main metabolites (S)-4-hydroxy-3-methoxymethamphetamine (HMMA) and (S)-3,4-dihydroxymethamphetamine (HHMA) were prepared for unequivocal identification of the differential enantioselective metabolism of these compounds as well as for its application in the analysis of biological samples. Capillary electrophoresis with cyclodextrin derivatives and a chemical correlation of (S)-MDMA, (S)-HMMA and (S)-HHMA has been performed to assign the absolute stereochemistry of major isomers in analytical standards enriched with such enantiomers.


Telework: systematic literature review and future research agenda.
Given the work and life conditions imposed by the 'new normal' Covid-19 era, a massive shift towards telework is expected and will likely continue long after the pandemic. Despite the resurgent interest in telework as an important aspect of ensuring business continuity, the literature base remains fragmented and variable. This study presents a taxonomical classification of literature on teleworking along with a comprehensive bibliography and future research agenda. To this aim, a systematic literature review methodology was adopted drawing on an evidence base of 40 articles published in high-ranking journals during the years 2000-2020. Findings capture key developments and synthesize existing areas of research focus. Important insights and gaps in the existing research are also pinpointed. The study may stimulate future research, represent a reference point for scholars interested in telework and at the same time provide an added advantage to managers for understanding crucial dimensions thereof.


Ocular features of CHARGE syndrome.
To detail the presence and severity of ocular and cranial nerve abnormalities found in individuals with CHARGE syndrome in a distinct geographic area. Nine individuals with CHARGE syndrome from Maritime Canada identified from a Canadian database were prospectively examined. Structural and sensorial defects associated with functional visual deficits were defined with ophthalmic and neurological evaluation. Consistent with current diagnostic criteria and the literature, colobomas were the major ophthalmic manifestation. These were typically bilateral chorioretinal colobomas involving the optic nerve. All subjects had bilateral severe sensorineural deafness (cranial nerve VIII), and 8 of 9 (89%) had facial nerve (cranial nerve VII) involvement (7 of 9 had unilateral involvement; 1 of 9 had bilateral involvement). Unique to this group of participants were the findings of anisometropia in 8 of the 9 (89%) patients, severe myopic astigmatism in 13 of the 18 eyes (72%), and limited elevation in adduction in 3 of 9 (33%) participants. Associated findings were strabismus, cataracts, microcornea, keratopathy, staphyloma, reduced stereopsis, superior visual field defects, and reduced visual acuity. The presence of coloboma plus another CHARGE feature warrants further investigation, including genetic screening for the CHD7 gene. Early recognition and management of sensory problems (visual, auditory, and vestibular) are crucial to ensure best psychomotor and cognitive development.


Oögenesis in a marine teleost, Blennius pholis L.
Oögenesis in the oviparous marine teleost, Blennius pholis L., is examined. Eleven developmental stages are identified by ultrastructural observations when changes in the distributions of the organelles and inclusions are described. An exogenous source for the protein yolk precursors is indicated, but less clear is the endogenous contribution. Changes in the follicle epithelium are described together with the formation of the zona which is considered to be follicular in origin. Two types of follicle cell are distinguished and these probably function differently in the process of zona formation. The zona becomes divided into the externa and interna, the latter probably resulting from the chemical ordering by disulphide bonding of the proteinaceous material of the former.


Central Auditory Processing Skills in Individuals with Takayasu Syndrome.
The current study was done to assess the central auditory processing in individuals with Takayasu's Syndrome. The Takayasu Syndrome is a rare disease, hence a case series was conducted with consecutive sampling. In this study, five patients diagnosed with Takayasu's syndrome were evaluated on the central auditory processing tests. The tests were administered using standard operating protocols. It was observed that the score on the dichotic digit test was drastically reduced suggesting difficulty in auditory integration skills in all the participants. There was an observed difficulty in speech perception in noise for all the participants indicating abnormal auditory closure ability. It was concluded that the reduced blood supply to the cortex would have caused a reduction in auditory skills. The current attempts to document the central processing disorder in a rare syndrome. The online version contains supplementary material available at 10.1007/s12070-023-03733-1.


Longer distance to specialized treatment centers does not adversely affect treatment intensity or outcomes in adult acute myeloid leukemia patients. A Danish national population-based cohort study.
Treatment of acute myeloid leukemia (AML) is widely centralized. Longer distances to a specialized treatment center may affect patients' access to curative-intended treatment. Especially during outpatient treatment, distance may also affect survival. The authors conducted a national population-based cohort study including all AML patients diagnosed in Denmark between 2000 and 2014. We investigated effects of distance (<10 kilometers [km; reference], 10-25, 25-50, 50-100, >100) to the nearest specialized treatment facility on the probability of receiving intensive chemotherapy, HSCT, and achieving a complete remission (CR) using logistic regression analysis (odds ratios; ORs). For overall survival, we used Cox proportional hazards regression (hazard ratios [HRs]) and adjusted (a) for relevant baseline characteristics. Of 2,992 patients (median age=68.5 years), 53% received intensive chemotherapy and 12% received low-dose chemotherapy outpatient regimens. The median distance to a specialized treatment center was 40 km (interquartile range=10-77 km). No impact of distance to specialized treatment centers was seen on the probability of receiving intensive chemotherapy (10-25 km, aOR=1.1 (CI=0.7-1.7), 25-50 km, aOR=1.1 (CI=0.7-1.7), 50-100 km, aOR=1.3 (CI=0.9-1.9), and >100 km, aOR=1.4 [CI=0.9-2.2]). Overall survival in patients regardless of therapy (<10 km, aOR=1.0 vs >100 km, aOR=1.0 [CI=0.9-1.2]), in intensive therapy patients, or in patients' choice of post-remission was not affected by distance to specialized treatment center. Distance to a transplant center also did not affect the probability of HSCT or survival post-HSCT. In Denmark, distance to a specialized treatment facility offering remission-induction chemotherapy and HSCT does not negatively affect access to curative-indented therapy, treatment-response, or survival in AML patients.


Fyn Kinase Controls Tau Aggregation In Vivo.
Alzheimer's disease (AD) is a proteinopathy exhibiting aggregation of β-amyloid (Aβ) as amyloid plaques and tau as neurofibrillary tangles (NFTs), whereas primary tauopathies display only a tau pathology. Aβ toxicity is mediated by Fyn kinase in a tau-dependent process; however, whether Fyn controls tau pathology in diseases that lack Aβ pathology remains unexplored. To address this, we generate the Tg/Fyn-/- mouse, which couples mutant tau overexpression with Fyn knockout. Surprisingly, Tg/Fyn-/- mice exhibit a near-complete ablation of NFTs, alongside reduced tau hyperphosphorylation, altered tau solubility, and diminished synaptic tau accumulation. Furthermore, Tg/Fyn-/- brain lysates elicit less tau seeding in tau biosensor cells. Lastly, the fibrillization of tau is boosted by its pseudophosphorylation at the Fyn epitope Y18. Together, this identifies Fyn as a key regulator of tau pathology independently of Aβ-induced toxicity and thereby represents a potentially valuable therapeutic target for not only AD but also tauopathies more generally.


Hormones and metabolites of arctic foxes (Alopex lagopus) in response to season, starvation and re-feeding.
Svalbard's arctic foxes experience large seasonal variations in light, temperature and food supply throughout the year, which may result in periods of starvation. The aim of this work is to investigate if there are seasonal variations in post-absorptive plasma thyroid hormones (free thyroxin (fT(4)), free triiodothyronine (fT(3)) and reverse triiodothyronine (rT(3))) and metabolites (free fatty-acids (FFA) and beta-hydroxybutyrate (beta-OHB)) with season and their response to starvation and re-feeding. The concentrations of post-absorptive free triiodothyronine were significantly higher in November than May, while those of thyroxin, reverse triiodothyronine, free fatty-acids and beta-hydroxybutyrate remained unchanged. Possible explanations for the seasonal variations in free triiodothyronine are discussed. There were no significant changes from post-absorptive concentrations of thyroxin and reverse triiodothyronine in starved and re-fed foxes. However, free triiodothyronine concentrations decreased during starvation and increased again with re-feeding both in May and November. Starvation induced high levels of free fatty acids in both May and November, indicating increased lipolysis. There was a significant increase in beta-hydroxybutyrate in November only, indicating that arctic foxes are capable of protein conservation during starvation.


Exploring the efficacy of 1-amino-cyclopropane-1-carboxylic acid (ACCA) as a natural compound in strengthening maize resistance against biotic and abiotic stressors: an empirical computational study.
This study aims to understand plant-bacteria interactions that enhance plant resistance to environmental stressors, with a focus on maize (Zea mays L.) and its vulnerability to various pathogenic organisms. We examine the potential of 1-amino-cyclopropane-1-carboxylic acid (ACCA) as a compound to boost maize's resilience against stressors and pathogens. With the growing global population and increased food demand, the study of endophytes, comprising bacteria and fungi, becomes crucial. They reside within plant tissues, affecting their hosts either beneficially or detrimentally. Agrobacteria are of specific interest due to their potential to contribute to developing strategies for plant resistance enhancement. We conducted exhaustive research on the defense-related proteins and mechanisms involved in maize-pathogen interactions. The efficacy of ACCA as a natural-compound that could enhance maize's resistance was examined. Our research indicates that ACCA, having a binding energy of -9.98 kcal/mol, successfully strengthens maize resistance against pathogenic assaults and drought stress. It plays a crucial protective role in maize plants as they mature, outperforming other ligands in its effectiveness to improve productivity and increase yield. Applying ACCA to maize plants has considerable potential in enhancing their resilience and tolerance to stress, proving to be an effective strategy to boost crop yield and productivity. This could help address the increasing global food demand. However, more research is needed to optimize ACCA application methods and to gain a comprehensive understanding of its long-term effects on maize cultivations and the environment.


Experimental study on the use of a chlorhexidine-loaded carboxymethylcellulose gel as antibacterial coating for hernia repair meshes.
Biomaterials with an antimicrobial coating could avoid mesh-associated infection following hernia repair. This study assesses the use of a chlorhexidine-loaded carboxymethylcellulose gel in a model of Staphylococcus aureus mesh infection. A 1% carboxymethylcellulose gel containing 0.05% chlorhexidine was prepared and tested in vitro and in vivo. The in vitro tests were antibacterial activity (S. aureus; agar diffusion test) and gel cytotoxicity compared to aqueous 0.05% chlorhexidine (fibroblasts; alamarBlue). For the in vivo study, partial abdominal wall defects (5 × 2 cm) were created in New Zealand white rabbits (n = 15) and inoculated with 0.25 mL of S. aureus (106 CFU/mL). Defects were repaired with a lightweight polypropylene mesh (Optilene) without coating (n = 3) or coated with a carboxymethylcellulose gel (n = 6) or chlorhexidine-loaded carboxymethylcellulose gel (n = 6). Fourteen days after surgery, bacterial adhesion to the implant (sonication, immunohistochemistry), host tissue incorporation (light microscopy) and macrophage reaction (immunohistochemistry) were examined. Carboxymethylcellulose significantly reduced the toxicity of chlorhexidine (p < 0.001) without limiting its antibacterial activity. While control and gel-coated implants were intensely contaminated, the chlorhexidine-gel-coated meshes showed a bacteria-free surface, and only one specimen showed infection signs. The macrophage reaction in this last group was reduced compared to the control (p < 0.05) and gel groups. When incorporated in the carboxymethylcellulose gel, chlorhexidine showed reduced toxicity yet maintained its bactericidal effect at the surgery site. Our findings suggest that this antibacterial gel-coated polypropylene meshes for hernia repair prevent bacterial adhesion to the mesh surface and have no detrimental effects on wound repair.


Nearly Panoramic Neuromorphic Vision with Transparent Photosynapses.
Neuromorphic vision based on photonic synapses has the ability to mimic sensitivity, adaptivity, and sophistication of bio-visual systems. Significant advances in artificial photosynapses are achieved recently. However, conventional photosyanptic devices normally employ opaque metal conductors and vertical device configuration, performing a limited hemispherical field of view. Here, a transparent planar photonic synapse (TPPS) is presented that offers dual-side photosensitive capability for nearly panoramic neuromorphic vision. The TPPS consisting of all two dimensional (2D) carbon-based derivatives exhibits ultra-broadband photodetecting (365-970 nm) and ≈360° omnidirectional viewing angle. With its intrinsic persistent photoconductivity effect, the detector possesses bio-synaptic behaviors such as short/long-term memory, experience learning, light adaptation, and a 171% pair-pulse-facilitation index, enabling the synapse array to achieve image recognition enhancement (92%) and moving object detection.


Time-resolved detection of hot electron-induced electrochemiluminescence of fluorescein in aqueous solution.
Strong electrogenerated chemiluminescence (ECL) of fluorescein is generated during cathodic pulse polarization of oxide-covered aluminum electrodes and the resulting decay of emission is so sluggish that time-resolved detection of fluorescein is feasible. The present ECL in aqueous solution is based on the tunnel emission of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons and hydroxyl radicals acting as redox mediators. The successive one-electron redox steps with the primary radicals result in fluorescein in its lowest excited singlet state. The method allows the detection of fluorescein (or its derivatives containing usable linking groups to biomolecules) over several orders of magnitude of concentration with detection limits well below nanomolar concentration level. The detection limits can still be lowered, e.g., by addition of azide or bromide ions as coreactants. The results suggest that the derivatives of fluorescein, such as fluorescein isothiocyanate (FITC), can be detected by time-resolved measurements and thus be efficiently used as electrochemiluminescent labels in bioaffinity assays.


An individual based, multidimensional approach to identify emotional reactivity profiles in inbred mice.
Despite extensive environmental standardization and the use of genetically and microbiologically defined mice of similar age and sex, individuals of the same mouse inbred strain commonly differ in quantitative traits. This is a major issue as it affects the quality of experimental results. Standard analysis practices summarize numerical data by means and associated measures of dispersion, while individual values are ignored. Perhaps taking individual values into account in statistical analysis may improve the quality of results. The present study re-inspected existing data on emotional reactivity profiles in 125 BALB/cJ and 129 mice, which displayed contrasting patterns of habituation and sensitization when repeatedly exposed to a novel environment (modified Hole Board). Behaviors were re-analyzed on an individual level, using a multivariate approach, in order to explore whether this yielded new information regarding subtypes of response, and their expression between and within strains. Clustering individual mice across multiple behavioral dimensions identified two response profiles: a habituation and a sensitization cluster. These retrospect analyses identified habituation and sensitization profiles that were similar to those observed in the original data but also yielded new information such as a more pronounced sensitization response. Also, it allowed for the identification of individuals that deviated from the predominant response profile within a strain. The present approach allows for the behavioral characterization of experimental animals on an individual level and as such provides a valuable contribution to existing approaches that take individual variation into account in statistical analysis.


Corrigendum to: Discriminating electrocardiographic responses to His-bundle pacing using machine learning [Cardiovascular Digital Health Journal 1 (2020) 11-20/5].
[This corrects the article DOI: 10.1016/j.cvdhj.2020.07.001.].


Demonstration of the ascorbate dependence of membrane-bound dopamine beta-monooxygenase in adrenal chromaffin granule ghosts.
Chromaffin granule ghosts from bovine adrenal medullae have been used to examine the ability of membrane-bound dopamine beta-monooxygenase to interact directly with intravesicular ascorbate and to investigate vectorial electron transfer from external ascorbate across the ghost membrane. Ghosts prepared by a modification of published procedures were shown to be fully active in both dopamine uptake and norepinephrine production. Dopamine uptake is dependent on the presence of a magnesium and ATP ionic complex, is abolished by reserpine, and reaches a steady-state level in the presence of dopamine beta-monooxygenase, ascorbate, catalase, and fumarate. Omission of ascorbate either inside or outside the ghosts greatly enhances dopamine accumulation, which reaches levels of approximately 30 nmol/mg under these conditions. Correspondingly, in the presence of all components, norepinephrine production reached approximately 100 nmol/mg in 30 min of incubation. Norepinephrine production was strictly magnesium-ATP-dependent, inhibited by either reserpine or dopamine beta-monooxygenase inactivation, and was markedly reduced when ascorbate was omitted from either inside or outside the ghosts. In the presence of limiting amounts of internal ascorbate, rapid norepinephrine production occurred which corresponded to the amount of initial ascorbate present, followed by a much slower endogenous norepinephrine production observable after complete depletion of internal ascorbate. The endogenous rate of norepinephrine production likely represents epinephrine-supported dopamine beta-monooxygenase turnover. Taken together, the data demonstrate that facile norepinephrine production by membrane-bound dopamine beta-monooxygenase occurs only when internal ascorbate is present, terminates upon depletion of internal ascorbate, and can only be sustained at a significant rate when reducing equivalents from external ascorbate are available.


Oxycodone does not affect placental circulatory physiology during the early first stage of labor-A randomized trial.
Opioids are used for pain relief during the first stage of labor. Oxycodone can cause maternal hypotension that may modify utero- and fetoplacental circulatory physiology. We hypothesized that maternal intravenous (i.v.) oxycodone has no detrimental effect on utero- and fetoplacental hemodynamics during the early first stage of labor. Twenty-two parturients requiring pain relief during the first stage of labor were randomized in a double-blinded and placebo-controlled study. By Doppler ultrasonography, both uterine artery (Ut) and umbilical vein (UV) volume blood flows (Q), Ut pulsatility index (PI), and Ut vascular resistance (RUt) were calculated. Blood flow velocity waveforms were obtained between uterine contractions. After baseline measurements, women received oxycodone 0.05 mg/kg or a placebo intravenous. Doppler ultrasonography was repeated up to 120 min after the first drug administration. The second dose of oxycodone 0.05 mg/kg was allowed at 60 min to all parturients with contraction pain ≥5/10. Maternal plasma samples were collected at each study phase and after delivery with umbilical cord plasma samples, to measure oxycodone concentrations. gov identifier (NCT no. NCT02573831). At baseline, mean QUt and QUV did not differ significantly between the placebo-first (478 mL/min and 57 mL/min/kg) and the oxycodone-first (561 mL/min and 71 mL/min/kg) groups. In addition, RUt and Ut PI were comparable between the groups. Following oxycodone at 60 min, mean QUt and QUV (714 mL/min and 52 mL/min/kg) were similar to the placebo-first (520 mL/min and 55 mL/min/kg) group. Furthermore, all the measured parameters were comparable to the baseline values. At 60 min after the first study drug administration, all the parturients in the placebo-first group needed intravenous oxycodone 0.05 mg/kg. At 120 min, we found no statistically significant change in any of the measured parameters. No significant correlation was found between maternal oxycodone concentration and QUt or QUV. Furthermore, newborn oxycodone concentration did not correlate with QUV. Oxycodone did not have any detrimental effect on either utero- or fetoplacental circulatory physiology during the early first stage of labor. Maternal plasma oxycodone did not correlate with utero- and fetoplacental hemodynamics. No correlation was found between newborn oxycodone concentration and fetoplacental hemodynamics.


Specificity of anti-DNA antibodies in SLE--II. Relative contribution of backbone, secondary structure and nucleotide sequence to DNA binding.
Fine specificity of a population of anti-DNA antibodies which bound both ssDNA and dsDNA with apparently equal affinity was studied in two SLE plasma. Sensitivity of DNA binding to increasing sodium chloride concentration indicated that electrostatic interactions occurred between antibody and phosphate moieties of DNA. Secondary nucleic acid structure was important to DNA binding as double-stranded synthetic deoxynucleotide polymers were more effective inhibitors than their substituent single-stranded polymers. Nucleotide bases were also found to play a role in recognition of DNA by these cross-reactive antibodies, as ssDNA binding was sensitive to increasing temperature which caused unstacking of the nucleotide bases. Differing patterns of reactivity with synthetic deoxynucleotide polymers with similar secondary structures but different nucleotide compositions further indicated the importance of nucleotide bases to dsDNA binding by cross-reactive anti-DNA antibodies in SLE plasma.


Considerations in placement of implants through existing split-thickness skin graft vestibuloplasty: a case report.
Many edentulous patients who previously have undergone vestibuloplasty with split-thickness skin grafting to improve denture stability and function seek additional help with dental implants. Problems encountered following the placement of four endosseous root-form dental implants in one such patient are presented. The anatomic and surgical considerations as well as methods available to circumvent these problems are discussed.


Acid-induced changes of brain protein buffering.
Excessive cellular acidosis is thought to enhance destruction of brain from ischemia. Protein denaturation may contribute to such injury although the behavior of brain proteins to acidosis is poorly defined. As a first approach to detect acid-induced changes in brain proteins and to characterize buffer content, homogenates were acidified for 20 min (as low as pH 3.1), returned to baseline pH (6.9), and then titrated. Titration curves show a significant (P less than 0.0001) and permanent increase in buffer content compared to controls when pH of acid exposure was 4.5-3.7 or less. Since acidity of pH 4.5 is rarely, if ever, achieved in vivo, protein denaturation from acidity alone is unlikely to account for necrosis of brain from ischemia.


A robust robotic high-throughput antibody purification platform.
Monoclonal antibodies (mAbs) have become the fastest growing segment in the drug market with annual sales of more than 40 billion US$ in 2013. The selection of lead candidate molecules involves the generation of large repertoires of antibodies from which to choose a final therapeutic candidate. Improvements in the ability to rapidly produce and purify many antibodies in sufficient quantities reduces the lead time for selection which ultimately impacts on the speed with which an antibody may transition through the research stage and into product development. Miniaturization and automation of chromatography using micro columns (RoboColumns(®) from Atoll GmbH) coupled to an automated liquid handling instrument (ALH; Freedom EVO(®) from Tecan) has been a successful approach to establish high throughput process development platforms. Recent advances in transient gene expression (TGE) using the high-titre Expi293F™ system have enabled recombinant mAb titres of greater than 500mg/L. These relatively high protein titres reduce the volume required to generate several milligrams of individual antibodies for initial biochemical and biological downstream assays, making TGE in the Expi293F™ system ideally suited to high throughput chromatography on an ALH. The present publication describes a novel platform for purifying Expi293F™-expressed recombinant mAbs directly from cell-free culture supernatant on a Perkin Elmer JANUS-VariSpan ALH equipped with a plate shuttle device. The purification platform allows automated 2-step purification (Protein A-desalting/size exclusion chromatography) of several hundred mAbs per week. The new robotic method can purify mAbs with high recovery (>90%) at sub-milligram level with yields of up to 2mg from 4mL of cell-free culture supernatant.


Characterization of Salmonella isolates from captive lizards.
Reptile-associated salmonellosis in humans is an increasing public health issue. This study aimed at characterizing Salmonella isolates from captive lizards and to compare them to human isolates. Salmonella was isolated from 25 of 33 cloacal and 47 of 79 faecal samples from captive lizards (75.8 and 59.5%, respectively). The strains belonged to 44 serotypes of subspecies I (27 serotypes), II (9), IIIb (3) and IV (5). Two strains, one of serotype Enteritidis and one of serotype Amsterdam, were resistant to nitrofurantoin. Invasion assays in Caco-2 cells were performed with 40 saurian isolates of subspecies I, 15 isolates of subspecies II, 4 strains of subspecies IIIb, 6 subspecies IV isolates and 17 human isolates of corresponding serotypes of subspecies I. Saurian isolates belonging to subspecies I invaded the Caco-2 cells to a higher extent than those from the other subspecies. The human isolates invaded the Caco-2 cells to a lesser degree compared to their saurian counterparts. In the same strains, the presence of virulence genes agfA, shdA, spvR, pefA and sopE was determined using PCR. Whereas agfA was detected in all strains, pefA was only detected in one saurian and in the human serotype Enteritidis strains. The spvR gene was detected in the same serotype Enteritidis strains and in 33% of the subspecies IV strains. The shdA gene was present in all the human isolates and in 86% of subspecies I saurian isolates. SopE was found in 17% of the human isolates, in 24% of the saurian subspecies I strains and in all of the subspecies IV strains.


Reliability of aortic MRI at 3 Tesla in patients with acute cryptogenic stroke.
To evaluate a new three dimensional (3D) MRI protocol for the reliable detection of aortic high risk plaques compared with transoesophageal echocardiography (TOE) and to test the reliability of additional MRI in stroke of undetermined aetiology. 74 acute stroke patients were examined by both TOE and MRI at 3 Tesla with special regard to aortic high risk plaques (ie, > OR = 4 mm, superimposed thrombi). ECG synchronised pre- and post-contrast T1 weighted 3D imaging (spatial resolution approximately 1 mm3) covering the thoracic aorta was employed. In plaques > OR = 3 mm, additional two dimensional T2 imaging and time resolved (CINE) imaging sequences were performed. Aetiology of brain ischaemia was classified according to modified TOAST (Trial of Org 10172 in Acute Stroke Treatment) criteria. Aortic high risk embolic sources detected by MRI in patients with cryptogenic stroke were evaluated. Differences in maximum aortic wall thickness for TOE and MRI were not statistically significant for all aortic segments. The overall number of high risk plaques detected by MRI (n = 74) was substantially higher compared with TOE (n = 47). Most noticeably, MRI identified aortic high risk pathologies in 8/26 (30.8%) patients with cryptogenic stroke after standard diagnostics, including TOE (n = 2: dissection or thrombus; n = 6: plaques > OR = 4 mm). Our results demonstrate the feasibility of this 3D MRI protocol for the reliable detection of aortic high risk plaques in acute stroke patients. Because of improved visualisation of the aortic arch and the detection of additional embolic sources not seen by standard diagnostics, this novel technique may become a valuable tool for future patients with cryptogenic stroke.


A comparison of plastic and plankton in the north Pacific central gyre.
The potential for ingestion of plastic particles by open ocean filter feeders was assessed by measuring the relative abundance and mass of neustonic plastic and zooplankton in surface waters under the central atmospheric high-pressure cells of the North Pacific Ocean. Neuston samples were collected at 11 random sites, using a manta trawl lined with 333 u mesh. The abundance and mass of neustonic plastic was the largest recorded anywhere in the Pacific Ocean at 334271 pieces km2 and 5114 g km2, respectively. Plankton abundance was approximately five times higher than that of plastic, but the mass of plastic was approximately six times that of plankton. The most frequently sampled types of identifiable plastic were thin films, polypropylene/monofilament line and unidentified plastic, most of which were miscellaneous fragments. Cumulatively, these three types accounted for 99% of the total number of plastic pieces.


Changes in expression of mesothelial BBS genes in 2D and 3D after lithium chloride and ammonium sulphate induction of primary cilium disturbance: a pilot study.
Malignant pleural mesothelioma (MPM), a rare and aggressive pleural tumor, has significant histological and molecular heterogeneity. Primary Cilium (PC), an organelle of emerging importance in malignancies, has been scarcely investigated in MPM. A critical molecular complex for the PC function is the BBSome and here we aimed at assessing its expression patterns in ordinary 2D and spheroid 3D cell cultures. A human benign mesothelial cell line (MeT-5A), MPM cell lines (M14K, epithelioid MPM; MSTO, biphasic MPM), and primary MPM cells (pMPM) were used. Primers specific for the human BBS1, 2, 4, 5, 7, 9, 18 transcripts were designed, and quantitative real-time PCR (qRT-PCR) was done with β-actin as the gene of reference. The relative gene expression across 2D and 3D cultures was analyzed by the expression factor (mean of 1/ΔCt values). With the 2-∆∆Ct method the gene expression fold changes were assessed from qRT-PCR data. Molecular changes using the PC-modulating drugs ammonium sulfate (AS) and lithium chloride (LC) were also determined. PC was present in all cells used in the study at approximately 15% of the observed area. BBSome transcripts were differentially expressed in different dimensions of cell culture (2D vs. 3D) in all cell lines and pMPM. Treatment with AS and LC affected the expression of the ciliary BBS2 and BBS18 genes in the benign as well as in the MPM cells. These data indicate distinct BBSome molecular profiles in human benign and MPM cells cultured in 2D and 3D dimensions and support the notion that PC genes should be investigated as potential MPM therapeutic targets.


The implications of bilingualism in the study and treatment of psychiatric disorders: a review.
This review of the data on bilinguals in relation to hypnosis, psychotherapy and psychotic disorders presents evidence for the bilateral representation of language in the cerebral hemispheres of bilinguals, and outlines possible links between the neuropsychology of psychotic disorders and the lateralization of language in these patients. The relationship between affective states and the retrieval of memories and language is also explored.


Persistent antigen and germinal center B cells sustain T follicular helper cell responses and phenotype.
T follicular helper (Tfh) cells provide help to B cells and are crucial for establishment of germinal center (GC) reactions, including production of high-affinity antibodies and generation of memory B cells and long-lived plasma cells. Here we report that the magnitude of the Tfh cell response was dictated by the amount of antigen and directly correlated with the magnitude of the GC B cell response. In addition, maintenance of the Tfh cell phenotype required sustained antigenic stimulation by GC B cells. In lymphopenic conditions, a strong and prolonged Tfh cell response led to bystander B cell activation, hypergammaglobulinemia, and production of poly- and self-reactive antibodies. These data demonstrate that antigen dose determines the size and duration of the Tfh cell response and GC reaction, highlight the transient nature of the Tfh cell phenotype, and suggest a link between overstimulation of Tfh cells and the development of dysregulated humoral immune responses.


A highly sensitive and reproducible HCV RNA hybridization method valuable for antiviral drug discovery.
Real-time RT-PCR and Northern blot are employed for the measurement of HCV RNA but suffer from multiple purification steps, high cost, and relatively large variability. In this study, a hybridization method for HCV RNA detection is described. This method does not need RNA purification, and is sensitive enough to detect HCV RNA present in replicon cellular lysates harvested from a single well of a 96-well plate. Fixation of RNA by UV cross-linking is crucial for this sensitivity. A linear relationship exists between hybridization signal and cell density ranging from 10(5) to as few as 300 cells per well. The signal-to-background ratio is greater than 40 and the Z factor is above 0.7. Using several known anti-HCV agents, dose-response curves and EC(50) values generated from hybridization were similar to those obtained from a luciferase assay. This method has been successfully applied to replicons of different HCV subtypes and hepatitis B virus in our laboratory. In summary, this hybridization assay is sensitive, highly reproducible, easy to handle, and a valuable tool for antiviral drug discovery.


Return to work improves quality of life and reduces psychosocial distress after radical cystectomy: data from a contemporary series of 230 German patients.
This study aims to evaluate health-related quality of life (HRQoL), psychosocial distress, and return to work (RTW) 2 years after radical cystectomy (RC) and inpatient rehabilitation (IR). The study relied on prospectively collected data for 842 patients, who underwent 3 weeks of IR after RC and creation of an ileal conduit (IC) or ileal neobladder (INB). Validated questionnaires surveyed patients on HRQoL and psychosocial distress (EORTC QLQ-C30, QSC-R10). Furthermore, employment status was evaluated. Regression was performed to identify predictors for HRQol, psychosocial distress, and RTW. Two-hundred and thirty patients were employed pre-surgery (77.8% INB, 22.2% IC). Patients with an IC suffered significantly more often from locally advanced disease (≥ pT3: 43.1% vs 22.9%; p = 0.004). Two years after surgery, 16.1% of patients had died (median days of survival 302 (IQR 204-482). Global HRQoL improved steadily, while high psychosocial distress was present in 46.5% of patients 2 years after surgery. Employment was reported by 68.2% of patients, of which 90.3% worked full-time. Retirement was reported by 18.5%. Multivariate logistic regression analysis identified age ≤ 59 years as the only positive predictor for RTW 2 years after surgery (OR 7.730; 95% CI 3.369-17.736; p < 0.001). Gender, surgical technique, tumor stage, and socioeconomic status did not influence RTW in this model. In multivariate linear regression analysis, RTW was identified as an independent predictor of better global HRQoL (p = 0.018) and lower psychosocial distress (p < 0.001), whereas younger patient age was identified as an independent predictor for higher psychosocial distress (p = 0.002). Global HRQoL and RTW are high among patients two years after RC. However, role and emotional, cognitive, and social functioning were significantly impaired, while high psychosocial distress persists in a material number of patients. Our study highlights how a successful RTW decreases psychosocial distress and increases QoL in patients after RC for urothelial cancer. Nonetheless, additional efforts by employers and healthcare providers are needed in aftercare after creation of an INB or IC.


Disulphide bonds in wheat gluten: cystine peptides derived from gluten proteins following peptic and thermolytic digestion.
Gluten from the wheat variety Rektor was extracted with 70% aqueous ethanol. The insoluble portion (whole glutenin) was partially hydrolysed with trypsin at pH 6.5 and separated on a Sephadex G25 column. The high molecular weight fraction 1 was further hydrolysed with pepsin at pH 2.0. To remove low molecular weight proteins, a portion of whole glutenin was extracted with dilute acetic acid. The residue (enriched glutenin), which contained mostly LMW and HMW subunits of glutenin, was hydrolysed with thermolysin at pH 6.5. The peptic and tryptic hydrolysates were separated on a Sephadex G25 column and the peptide fractions with the highest cystine content were separated further by reversed-phase high-performance liquid chromatography (RP-HPLC). Cystine peptides were detected by differential chromatography (RP-HPLC prior to and after reduction of disulphide bonds) and then isolated by preparative RP-HPLC. After reduction, cysteine peptides were alkylated and analysed for their amino acid sequence. Altogether, 19 cystine peptides were characterized and assigned to known sequences of gluten proteins; 16 peptides confirmed the positions of disulphide bonds present in LMW subunits and gamma-gliadins, as described previously. For the first time, a cystine peptide has been isolated, representing an intermolecular disulphide bond between the y-type of HMW and LMW subunits. Furthermore, a cystine peptide was assigned to gamma-gliadins; thus, all cysteine residues of gamma-gliadins are documented by at least one cystine peptide. One peptide analysed came from the alpha-amylase inhibitor CM 16. Altogether the results indicate that the intramolecular linkages of gluten proteins are not formed at random, but are strongly directed.(ABSTRACT TRUNCATED AT 250 WORDS)


[Atherogenesis and immune inflammation].
A universal approach to atherosclerosis pathogenesis is discussed according to which humoral factors, first of all modified lipoproteins and cells of the vascular wall, are considered as an entity. Although the arterial wall is estimated as a target organ realizing leading pathogenetic factors, the process in the arterial wall is a trigger of cell reactions resulting in the formation of an atherosclerotic plaque. Immune inflammation developing in the vascular wall as a response to deposition or formation of the autoimmune complex that includes a modified lipoprotein as an antigen, is regarded as an integral part of atherogenesis. Cytokines produced by the activated macrophages and lymphocytes purposefully affect parietal cells promoting the formation of foam cells and phenotypic modification of GMK with their production of the components of the connective tissue matrix.


Vertebral artery dissections with and without cervical spine fractures: Analysis of 291 patients.
Vertebral artery dissections occur when tears in the intimal layer of the vertebral artery and are associated with trauma, infection, and spontaneous etiologies. We aimed to identify differences in predisposing factors and outcomes in vertebral artery dissections associated with cervical spine fractures compared to those not associated with cervical spine fractures. We conducted a retrospective chart review of patients with vertebral artery dissections who presented to our institution at the time of dissection and had a minimum of 3 month follow-up and collected data on demographics, event characteristics, treatments, and outcomes in the form of modified Rankin scale scores. In total, 291 patients with VAD were included in this study. Thirty-nine patients with VADs had associated fracture, while 252 patients had VADs without fracture. VAD patients with associated cervical fractures were more likely to be male (p < 0.001), have a greater number of comorbid conditions (p < 0.01), be smokers (p = 0.045), or have violence (p < 0.001) or motor vehicle accidents (p < 0.001) as the cause of their VADs. VAD patients with associated cervical fractures were less likely to have associated aneurysms or pseudoaneurysms (p = 0.002). VAD patients with associated cervical fractures were more likely to have higher mRS at discharge from the hospital (p < 0.001), 3 month follow-up (p < 0.001), and last follow-up (p < 0.001). Cervical spine fracture is likely the primary driver of poor neurological outcomes following vertebral artery dissection with associated cervical spine fracture.


A reappraisal of the role of cyclic AMP in the physiological action of glucagon.
Effects of the metabolic hormone glucagon can be physiological or supraphysiological, based on agonist concentration and the mediating cellular signal. The threshold concentration (TC) for activating the AC/cAMP signal pathway in liver is ≥ 100 pM. By contrast, mean plasma concentrations are around 20-45 pM, depending on the vascular bed. Accordingly, effects produced at TCs below 100 pM are physiological and mediated by cellular signal pathways other than AC/cAMP. Effects generated at concentrations above 100 pM are supraphysiological, often mediated by simultaneous activation of cAMP-independent and -dependent pathways. Physiological responses, and their established or implicated signal pathways, include stimulation of: glucose mobilization, fatty acid oxidation, and urea synthesis in liver (PLC/IP3/Ca2+/CaM); lipolysis in white and brown adipose tissue and oxygen consumption in brown adipose of the rat but not in humans (PLC/IP3/Ca2+/CaM); renal potassium and phosphate excretion in rodents and GFR in humans (signal undetermined); and glucose utilization in rat heart (PI3K/akt). Supraphysiological responses involve the AC/cAMP pathway and include: enhanced stimulation of glucose mobilization and stimulation of urea synthesis in liver; further stimulation of white and brown adipose lipolysis and thermogenesis in brown adipose tissue; stimulation of renal Cl- transport; and increased rat heart contractility. The AC/cAMP pathway is likely recruited when plasma glucagon rises above 100 pM during periods of elevated metabolic stress and systemic glucose demand, such as in the early neonate or strenuously exercising adult. The current cAMP-centered model should therefore be reconsidered and replaced with one that places more emphasis on cAMP-independent pathways.


Effects of amiodarone and dronedarone on voltage-dependent sodium current in human cardiomyocytes.
Effect of Dronedarone on Cardiac Na Current. Amiodarone (AM) is a highly effective antiarrhythmic agent used in the management of both atrial and ventricular arrhythmias. Its noniodinated analogue dronedarone (SR) may have fewer side effects than AM. In this study, we compared the effects of AM and SR on the sodium current I(Na) in human atrial myocytes. INa was studied with the whole-cell, patch clamp technique. Both AM and SR induced a dose-dependent inhibition of I(Na) recorded at -40 mV from a holding potential of -100 mV. AM inhibited I(Na) by 41%+/- 11% (n = 4) at 3 microM, and by 80%+/- 7% (n = 5) at 30 microM. SR produced more potent block, inhibiting INa significantly at only 0.3 microM (23%+/- 10%, n = 4) and completely (97%+/- 4%, n = 4) at 3 microM. Both AM and SR had only moderate effects on voltage-dependent properties of I(Na) (current-voltage relationship, availability for activation) and had no effect on the current decay kinetics. Both AM and SR inhibit I(Na) significantly in single human atrial cells, showing that the two drugs have Class I antiarrhythmic properties. The acute effects of SR are more potent than those of AM. The study supports the idea that the iodinated form of the molecule has no part in the acute effect of AM on Na+ channels.


Characterisation and evaluation of UK websites on attention deficit hyperactivity disorder.
To identify, characterise and evaluate UK websites providing information about attention deficit hyperactivity disorder (ADHD) and its pharmacological management. Cross-sectional survey of websites identified by entering "ADHD" into five search engines. 48 websites. Each website was scored against 26 criteria using a bespoke instrument to evaluate (a) quality of information on the disorder and its drug treatment and (b) physical characteristics of the site. Most sites (n = 22) were hosted by charities and support groups, 12 were by commercial organisations, nine were from government or professional bodies, and five were categorised as miscellaneous. Mean total scores per host category ranged from 18.8 to 21 out of 46, with mean (SD) scores of 5.5 (4.2) out of 28 for content and 14.8 (3.0) out of 18 for physical properties. The government/professional sites scored highest for both content and physical properties. Descriptions of the disorder and its drug treatment were poor and lacking in detail. Although most sites mentioned stimulants, only eight discussed atomoxetine and described how both types of drug worked. Ten sites provided detailed information about side effects. The role of different stimulant brands and formulations was discussed on six sites. Authorship details were generally vague. Physical properties related to navigation and layout performed well. Only four sites used language deemed suitable for consumer-orientated health information. Information on UK websites about drug treatment for ADHD is basic and incomplete. Websites by government and professional bodies perform better than those in other categories.


Investigating genetic polymorphism in E. histolytica isolates with distinct clinical phenotypes.
Amoebiasis is an infection caused by enteric protozoa, most commonly Entamoeba histolytica, and is globally considered a potentially severe and life-threatening condition. To understand the impact of the parasite genome on disease outcomes, it is important to study the genomes of infecting strains in areas with high disease prevalence. These studies aim to establish correlations between parasite genotypes and the clinical presentation of amoebiasis. We employ a strain typing approach that utilizes multiple loci, including SREHP and three polymorphic non-coding loci (tRNA-linked array N-K2 and loci 1-2 and 5-6), for high-resolution analysis. Distinct clinical phenotype isolates underwent amplification and sequencing of studied loci. The nucleotide sequences were analysed using Tandem Repeats Finder to detect short tandem repeats (STRs). These patterns were combined to assign a genotype, and the correlation between clinical phenotypes and repetitive patterns was statistically evaluated. This study found significant polymorphism in the size and number of PCR fragments at SREHP and 5-6 locus, while the 1-2 locus and NK2 locus showed variations in PCR product sizes. Out of 41 genotypes, two (I6 and I41) were significantly associated with their respective disease outcomes and were found in multiple isolates. We observed that I6 was linked with a symptomatic outcome, with a statistically significant p-value of 0.0183. Additionally, we found that I41 was associated with ALA disease outcome, with a p-value of 0.0089. Our study revealed new repeat units not previously reported, unveiling the genetic composition of E. histolytica strains in India, associated with distinct disease manifestations.


C-reactive protein and prognosis in women and men with coronary artery disease after percutaneous coronary intervention.
Sex-based differences in the association between C-reactive protein (CRP) and cardiovascular events in patients with coronary artery disease (CAD) are incompletely investigated. We investigated whether there are gender differences in the association between CRP and outcome in patients with CAD after percutaneous coronary intervention (PCI). This study included 13,170 consecutive patients with CAD: 10,098 men and 3072 women. CRP was measured on admission in all patients. The primary outcome was 1-year mortality. CRP level (median [25th-75th percentiles]) was higher in women than in men (3.08 [1.30-8.37] mg/L vs 2.30 [0.92-6.47] mg/L; P<0.001). CRP was >3mg/L in 4250 men (42.1%) and 1554 women (50.6%; P<0.001). One-year mortality was 4.9% (n=641 deaths). Deaths occurred in 318 men with CRP >3mg/L and 122 men with CRP ≤3mg/L (mortality estimates 7.7% and 2.1%, P<0.001) and in 154 women with CRP >3mg/L and 47 women with CRP ≤3mg/L (mortality estimates 10.1% and 3.2%, P<0.001). After adjustment in the Cox model, CRP was associated with increased risk of mortality in women (adjusted hazard ratio [HR]=1.03, 95% confidence interval [CI] 1.01-1.04, P<0.001 for each 5mg/L increase) and in men (adjusted HR=1.02 [1.01-1.03], P<0.001, for each 5mg/L increase). CRP predicted mortality with an area under the receiver-operating characteristic curve =0.721, [0.683-0.760] in women and 0.732, [0.707-0.757] in men (P=0.659). Elevated CRP levels provide similar prognostic information in men and women with CAD after PCI which is independent and supplementary to that provided by conventional cardiovascular risk factors.


Generation of autologous tumor-specific T cells for adoptive transfer based on vaccination, in vitro restimulation and CD3/CD28 dynabead-induced T cell expansion.
Adoptive cell transfer (ACT) of in vitro expanded autologous tumor-infiltrating lymphocytes (TIL) has been shown to exert therapeutic efficacy in melanoma patients. We aimed to develop an ACT protocol based on tumor-specific T cells isolated from peripheral blood and in vitro expanded by Dynabeads® ClinExVivo™CD3/CD28. We show here that the addition of an in vitro restimulation step with relevant peptides prior to bead expansion dramatically increased the proportion of tumor-specific T cells in PBMC-cultures. Importantly, peptide-pulsed dendritic cells (DCs) as well as allogeneic tumor lysate-pulsed DCs from the DC vaccine preparation could be used with comparable efficiency to peptides for in vitro restimulation, to increase the tumor-specific T-cell response. Furthermore, we tested the use of different ratios and different types of Dynabeads® CD3/CD28 and CD3/CD28/CD137 T-cell expander, for optimized expansion of tumor-specific T cells. A ratio of 1:3 of Dynabeads® CD3/CD28 T-cell expander to T cells resulted in the maximum number of tumor-specific T cells. The addition of CD137 did not improve functionality or fold expansion. Both T-cell expansion systems could generate tumor-specific T cells that were both cytotoxic and effective cytokine producers upon antigen recognition. Dynabeads®-expanded T-cell cultures shows phenotypical characteristics of memory T cells with potential to migrate and expand in vivo. In addition, they possess longer telomeres compared to TIL cultures. Taken together, we demonstrate that in vitro restimulation of tumor-specific T cells prior to bead expansion is necessary to achieve high numbers of tumor-specific T cells. This is effective and easily applicable in combination with DC vaccination, by use of vaccine-generated DCs, either pulsed with peptide or tumor-lysate.


Sequence-non-specific effects generated by various types of RNA interference triggers.
RNA interference triggers such as short interfering RNA (siRNA) or genetically encoded short hairpin RNA (shRNA) and artificial miRNA (sh-miR) are widely used to silence the expression of specific genes. In addition to silencing selected targets, RNAi reagents may induce various side effects, including immune responses. To determine the molecular markers of immune response activation when using RNAi reagents, we analyzed the results of experiments gathered in the RNAimmuno (v 2.0) and GEO Profiles databases. To better characterize and compare cellular responses to various RNAi reagents in one experimental system, we designed a reagent series in corresponding siRNA, D-siRNA, shRNA and sh-miR forms. To exclude sequence-specific effects the reagents targeted 3 different transcripts (Luc, ATXN3 and HTT). We demonstrate that RNAi reagents induce a broad variety of sequence-non-specific effects, including the deregulation of cellular miRNA levels. Typical siRNAs are weak stimulators of interferon response but may saturate the miRNA biogenesis pathway, leading to the downregulation of highly expressed miRNAs, whereas plasmid-based reagents induce known markers of immune response and may alter miRNA levels and their isomiR composition.


The effects of community-based interventions on the uptake of selected maternal and child health services: experiences of the IMCHA project in Iringa Tanzania, 2015-2020.
Maternal and child health (MCH) improvement has been prioritised in resource-constrained countries. This is due to the desire to meet the global sustainable development goals of achieving a maternal mortality rate of 70 per 100000 live births by 2030. The uptake of key maternal and child health services is crucial for reducing maternal and child health mortalities. Community-Based Interventions (CBIs) have been regarded as among the important strategies to improve maternal and child health service uptake. However, a paucity of studies examines the impacts of CBIs and related strategies on maternal and child health. This paper unveils the contribution of CBIs toward improving MCH in Tanzania. Convergent mixed method design was employed in this study. Questionnaires were used to examine the trajectory and trend of the selected MCH indicators using the baseline and end-line data for the implemented CBI interventions. Data was also collected through in-depth interviews and focus group discussions, mainly with implementers of the interventions from the community and the implementation research team. The collected quantitative data was analysed using IBM SPSS, while qualitative data was analysed thematically. Antenatal care visits increased by 24% in Kilolo and 18% in Mufindi districts, and postnatal care increased by 14% in Kilolo and 31% in Mufindi districts. Male involvement increased by 5% in Kilolo and 13% in Mufindi districts. The uptake of modern family planning methods increased by 31% and 24% in Kilolo and Mufindi districts, respectively. Furthermore, the study demonstrated improved awareness and knowledge on matters pertaining to MCH services, attitude change amongst healthcare providers, and increased empowerment of women group members. Community-Based Interventions through participatory women groups are vital for increasing the uptake of MCH services. However, the success of CBIs depends on the wide array of contextual settings, including the commitment of implementers of the interventions. Thus, CBIs should be strategically designed to enlist the support of the communities and implementers of the interventions.


Prevalence of and factors related to anemia among Japanese adult women: Secondary data analysis using health check-up database.
The issue of anemia is important in terms of a woman's preconception health. This study aimed to conduct an exploratory investigation of the prevalence of and factors related to anemia in non-pregnant Japanese women. Secondary data analysis was conducted using a database of women aged 20-49 years old who had attended an annual health check-up at a hospital in Tokyo (n = 10,598). A multiple logistic regression analysis was performed to identify factors related to anemia in two age groups: women aged 34 and under and those aged 35 and over. Anemia was defined as hemoglobin concentration levels <12.0 g/dL. The overall proportion of women with Hb <12.0 g/dL was 17.1%. Women aged 35 and over with a current medical history of uterine myoma were found to be at a higher risk of anemia. Women aged 35 and over who were overweight had a lower risk of anemia than women with normal weights. Current and past smoking habits affected hemoglobin levels among women aged 35 and over. In both age groups, those who drank alcohol habitually were at a lower risk than those who did not. Related factors of anemia should take into consideration a woman's age and lifestyle.


Epidermal penetration and protoporphyrin IX formation of two different 5-aminolevulinic acid formulations in ex vivo human skin.
Photosensitizer formation and epidermal penetration depth represent basic predictors of drug efficacy in dermatological Photodynamic Therapy (PDT). Different drug formulations and application standards are used to perform PDT in clinical practice. Thus, we developed a human ex vivo model suitable to explore drug permeation in human skin and compared in 10 patients the penetration of nanoemulsion formulation (BF-200 ALA) with that of a 20% ALA cream formulation frequently used in clinical practice. Protoporphyrin IX (PpIX) formation was assessed according to different durations of incubation with both preparations. BF-200 ALA led to more intense PpIX fluorescence than the 20% ALA cream formulation as assessed by fluorescence microscopy: after 12h of incubation, total measured fluorescence was at 101,995 fluorescence units with BF-200 ALA and 40,960 fluorescence units with 20% ALA cream, respectively. This could be reproduced using quantitative fluorimetric measurements in tissue lysates. After the clinically relevant incubation time of 3h the PpIX concentration induced by BF-200 ALA was more than three-fold higher than that induced by the 20% ALA formulation (7.1±5.5 and 1.9±1.8nmol/l, p<0.05, Mann-Whitney U-test) and four-fold higher after 12h (30.0±4.6 and 6.7±2.0nmol/l, p<0.01, Mann-Whitney U-test). In spite of the 50% lower ALA content BF-200 ALA triggers significantly higher PpIX concentrations than the 20% ALA formulation, indicating that clinical efficacy with BF-200 ALA may be higher. Moreover, the ex vivo eyelid skin model may represent a useful tool to investigate drug permeation in human skin.


Sacubitril/Valsartan in Dialysis Patients: Update on Current Perspectives.
Sacubitril/Valsartan is a combination of neprilysin inhibitor and angiotensin II receptor blocker that proved its own efficacy and safety in heart failure patients to ameliorate cardiovascular morbidity and mortality compared to angiotensin II-converting enzyme inhibitors alone. However, end-stage renal disease patients have not been included in the randomized controlled trials, so the beneficial effects as well as the risk profile of this association remain still undefined in these patients. Only observational studies on this drug association have been carried out in end-stage renal disease patients investigating mostly biohumoral or echocardiographic markers. Therefore, its application is still controversial and not free of complications due to the potential risk of hypotension and hyperkaliemia. The efficacy to improve biohumoral markers and cardiac function in dialysis patients and the potential application especially in those patients with severe and resistant hypertension and/or left ventricular dysfunction could be crucial in end-stage renal disease patients. Ongoing long-term randomized controlled trials should thoroughly define the effective benefits and/or adverse effects in patients on substitutive treatment.


Mixed shell mesoporous silica nanoparticles for controlled drug encapsulation and delivery.
Smart mesoporous silica nanoparticles (MSNs) with mixed polymeric shell (MS-MSNs) were prepared to realize controlled encapsulation and responsive delivery of anticancer drugs. Two kinds of polymers, including nonthermoresponsive poly(ethylene glycol) and thermoresponsive poly(N-isopropyl acrylamide), were grafted onto the outlets of the MSNs through acidic liable Schiff base bonds. Poly(N-isopropyl acrylamide) chains could control the release rate of drugs through phase transition, while poly(ethylene glycol) chains could maintain the colloid stability of MSNs. Drugs can be released through the gradual hydrolysis of Schiff base bonds in tumor acidic environment. The MS-MSNs gave consideration to both the responsiveness and stability of carriers, and could realize the release of drugs as much as possible in tumor tissues.


Nature exposure might be the intervention to improve the self-regulation and skilled performance in mentally fatigue athletes: A narrative review and conceptual framework.
Due to causing inability of self-regulation (ego depletion) and executive functions such as directed attention and visual searching for relevant information (e.g., the ball location and the position of teammates), mental fatigue impairs skilled performance in various sports. On the other hand, natural scenes could improve directed attention, which may considerably benefit visual searching ability and self-regulation. However, nature exposure as a potential intervention to improve skilled performance among mentally fatigued athletes has not been discussed thoroughly. To propose the potential intervention for the impairment of skilled performance among mentally fatigued athletes and generate a framework for future studies. A narrative review was applied to search broadly across disciplines, retrieving literature from several databases (PubMed, Web of Science, Scopus, and EBSCOhost). Thirty-three works of literature including 39 experiments (mental fatigue 15; ego depletion 5; and nature exposure 19) were obtained. Finally, a conceptual framework was built regarding the effect of nature exposure intervention on skilled performance in athletes for future studies. Three theories (the psychobiological model of exercise performance, the strength model of self-regulation, and attention restoration theory) could be potentially integrated to be a conceptual framework and explain the mechanism of preventing prior mental exertion (e.g., mental fatigue and ego depletion). Future studies could examine more on the duration of the intervention.


Mapping of the World Health Organization Quality of Life Brief (WHOQOL-BREF) to the EQ-5D-5L in the General Thai Population.
This study aimed at mapping the World Health Organization Quality of Life Brief (WHOQOL-BREF) and the EQ-5D-5L in the general Thai population and to determine the impact on the incremental cost-utility ratio (ICUR) through five hypothetical scenarios. A total of 1,200 Thai participants were randomly allocated into the 'estimation' and 'validation' groups. A curve estimation with nine regression models was performed to identify the best-fit regression model of significant WHOQOL-BREF dimension scores for the EQ-5D-5L index score predictions in the estimation group. The identified model was then used for the calculation of the predicted EQ-5D-5L index scores in the validation group. The percentage change from the hypothetical base-case scenario with predefined parameters was used to determine the impact on the ICUR. An inverse model was the best-fit regression model to predict the EQ-5D-5L index scores. The absolute difference between the predicted and observed index scores was 0.064, and the percentage of the sample that was mispredicted by ≥ 0.05 and ≥ 0.1 was 43.8% and 16.8%, respectively. Moreover, the percentage change in ICUR ranged between 0.13 and 1.84% from the hypothetical base-case scenario. An inverse relationship between the studied scores was identified. The minimal impact on the ICUR suggests that the Health Utility Index of the mapped equation can be applied to economic analyses.


Impact of sequencing depth and read length on single cell RNA sequencing data of T cells.
Single cell RNA sequencing (scRNA-seq) provides great potential in measuring the gene expression profiles of heterogeneous cell populations. In immunology, scRNA-seq allowed the characterisation of transcript sequence diversity of functionally relevant T cell subsets, and the identification of the full length T cell receptor (TCRαβ), which defines the specificity against cognate antigens. Several factors, e.g. RNA library capture, cell quality, and sequencing output affect the quality of scRNA-seq data. We studied the effects of read length and sequencing depth on the quality of gene expression profiles, cell type identification, and TCRαβ reconstruction, utilising 1,305 single cells from 8 publically available scRNA-seq datasets, and simulation-based analyses. Gene expression was characterised by an increased number of unique genes identified with short read lengths (<50 bp), but these featured higher technical variability compared to profiles from longer reads. Successful TCRαβ reconstruction was achieved for 6 datasets (81% - 100%) with at least 0.25 millions (PE) reads of length >50 bp, while it failed for datasets with <30 bp reads. Sufficient read length and sequencing depth can control technical noise to enable accurate identification of TCRαβ and gene expression profiles from scRNA-seq data of T cells.


Dioctophyme renale: in vitro culture, production of excretory and secretory antigens, and their use in immunodiagnosis.
The nematode Dioctophyme renale affects mainly the right kidney of domestic and wild mammals, in addition to having zoonotic potential. Therefore, this study aimed to produce excretion and secretion antigens of adult D. renale (DES) to diagnose canine dioctophimosis. To obtain the excretion and secretion antigens (DES), five D. renale adults were maintained for three weeks in supplemented RPMI medium with weekly supernatant collections. After the DES concentration, 2200 mL of the collected supernatant was used, which was centrifuged, followed by two filtrations and dialyzed, resulting in 12.5 mL of DES with a protein concentration of 0.59 mg/mL. The polyacrylamide gel electrophoresis (SDS-PAGE) of the DES antigen showed fractions with molecular weights ranging from 30 to 250 Kda. In the indirect ELISA with the DES antigen, the mean absorbance of the positive sera (38) was 0.839 ± 0.267, and the mean of the negative control sera (7) was 0.208 ± 0.083; the specificity and sensitivity of the ELISA were 100 and 97.4 %, respectively, being as effective as the surgical method in the presence of viable parasites. Thus, for the first time, this study describes the production of excretion and secretion antigens of adult D. renale and standardizes an indirect ELISA to diagnose dioctophimosis.


Understanding the prevalence and manifestation of anxiety and other socio-emotional and behavioural difficulties in children with Developmental Language Disorder.
It is well-documented that children with Developmental Language Disorder (DLD) have a higher likelihood of experiencing anxiety, as well as other socio-emotional and behavioural (SEB) difficulties. Despite this, there is little consensus as to how these difficulties manifest. This study aims to understand the prevalence of broader SEB difficulties and anxiety, informing intervention development by understanding the relationships between them. A mixed-methods, case-control study was conducted. First, an online survey was completed by 107 parents of either children with DLD ("DLD sample"; n = 57) or typically developing children ("typical sample"; n = 50), aged 6-12 years old. Binary SEB statements informed by previous qualitative work (e.g. "my child requires routine/sameness"; "my child has frequent tantrums") provided an insight into the prevalence of SEB difficulties in both DLD and typical samples. Validated measures of anxiety, emotion regulation, intolerance of uncertainty, insistence on sameness, family stress and coping mechanisms were also collected. Correlation and mediation analyses were run using these validated measures to understand the manifestation of anxiety in children with DLD in more detail. Qualitative interviews were then carried out with a select panel of survey respondents (n = 4). The DLD sample scored significantly higher on all binary SEB statements than the typical sample: experiencing anxiety (80.7%, p < .05), requiring routine and sameness (75.4%, p < .001) and emotional dysregulation (75.4%; p < .001) were the most common difficulties reported for children with DLD. Using the validated scales, family stress and coping mechanisms were found to only correlate with the manifestation of anxiety in the typical group, not the DLD group. "Intolerance of uncertainty" and "insistence on sameness" were found to fully mediate the relationship between DLD diagnosis and symptoms of anxiety. Parent's interviews provided contextual support for the analysis, as well as highlighting sensory sensitivities as a focus for future research. Parents of children with DLD appear to cope well with their children's complex SEB needs. Intervention focussing on intolerance of uncertainty may help the management of difficulties with anxiety. Behaviours such as insistence on sameness should be investigated further, as potential indicators for anxiety amongst children with DLD.


Effect of a high fat/cholesterol diet with or without eicosapentaenoic acid on plasma lipids, lipoproteins and lipid transfer protein activity in the marmoset.
Marmosets fed a diet supplemented with 0.2% cholesterol and 10% sheep fat (by weight) developed hypercholesterolemia with a 4-fold increase in plasma cholesterol (4.28 +/- 0.57-16.38 +/- 4.22 mmol/l, mean +/- SD, P less than 0.001). This was due mainly to a 5-fold increase in the intermediate density lipoprotein (IDL) and low density lipoprotein (LDL) fraction (d = 1.006-1.063 g/ml). The proportion of plasma cholesterol in high density lipoproteins (HDL) decreased from 56% to 25% although HDL cholesterol increased from 2.40 +/- 0.42 to 4.09 +/- 0.92 mmol/l (P less than 0.001), and HDL particle radius increased from 5.10 +/- 0.18 nm to 6.06 +/- 0.73 nm (P less than 0.05). Plasma lipid transfer protein (LTP) activity increased 2.5-fold in whole plasma and 2-fold in lipoprotein-deficient plasma. The atherogenic lipoprotein profile was attenuated by adding 0.8% eicosapentaenoic acid (EPA, 20:5 n - 3, as the ethyl ester) to the atherogenic diet. Plasma cholesterol increased only 55% to 6.64 +/- 2.55 mmol/l with only an 80% increase in lipoproteins in the d = 1.006-1.063 g/ml fraction and a more favourable proportion of plasma cholesterol in HDL (44%) than without EPA. LTP activity was reduced to 1.7-fold above control in whole plasma by addition of EPA to the atherogenic diet. There was a positive correlation between plasma cholesterol and LTP activity in whole plasma (r = 0.89, P less than 0.001) and in lipoprotein-deficient plasma (r = 0.67, P less than 0.001). EPA therefore attenuated some of the adverse effects of a 0.2% cholesterol, 10% sheep fat diet on plasma lipids and lipoproteins and induced a less atherogenic profile.


Safety and efficacy of atorvastatin in heart transplant recipients.
Pravastatin and simvastatin prolong survival and reduce transplant-related coronary vasculopathy, although low-density lipoprotein (LDL) lowering with these agents is only modest. The objective of this study was to assess the safety of moderate dose atorvastatin and its efficacy when prior treatment with another statin had failed to lower LDL to < 100 mg/dl. Data from 185 patients were retrospectively evaluated for adverse events, duration of exposure (person-days), and the mean atorvastatin dose exposure. Changes in lipid parameters, and prednisone and cyclosporine doses were determined. 48 patients received atorvastatin for 24,240 person-days at a mean dose exposure of 21 +/- 10 mg. Rhabdomyolysis, myositis, myalgias, and hepatotoxicity occurred in 0, 2, 2, and 0 patients, respectively. All events occurred at the 10-mg dose, within the first 3 months, and were rapidly reversible with atorvastatin discontinuation. Thirty-four patients evaluable for efficacy analyses had a pre-atorvastatin LDL of 145 +/- 38 mg/dl on the following statins: pravastatin (n = 30, 40 +/- 0mg), fluvastatin (n = 3, 33 +/- 12 mg), simvastatin (n = 1, 40 mg). After atorvastatin (21 +/- 9 mg/day) for 133 +/- 67 days, LDL was reduced to 97 +/- 24 mg/dl (relative reduction 31 +/- 20%, p < 0.0001). At the end of the observation period (418 +/- 229 days, atorvastatin final dose 24 +/- 14 mg/day), LDL was further decreased to 88 +/- 23 mg (relative reduction 37 +/- 17%, p < 0.0001). Atorvastatin, when used at moderate doses and with close biochemical and clinical monitoring, appears to be safe and is effective in aggressively lowering LDL in heart transplant recipients when treatment with other statins has failed to achieve LDL goals.


Investigation of various adsorbents for their ability to bind aflatoxin B1.
The contamination of animal feed with mycotoxins represents a worldwide problem for the animal industry. The most applied method for protecting animals against aflatoxicosis is the utilization of clay minerals. In the course of a research project adsorption experiments were performed in buffer solutions in order to evaluate the ability to bind Aflatoxin B1 (AfB1) at various pH-values. In order to investigate the strength of binding, the chemisorption index was calculated. Isothermal analysis was used to determine the values for the maximum adsorption capacity. Adsorption experiments in simulated gastrointestinal fluid and real gastric juice were carried out. Furthermore binding capability of the materials regarding selected vitamins was examined. Special attention was paid to the formation of AfB2a during experimental conditions. Based on the obtainedin vitro results, highly promising sorbent materials were ranked for furtherin vivo studies. Some adsorbing bentonites were also analysed mineralogically, but the results did not indicate which smectite property influences the adsorption process for AfB1.


Rafoxanide promotes apoptosis and autophagy of gastric cancer cells by suppressing PI3K /Akt/mTOR pathway.
Rafoxanide is commonly used as anti-helminthic medicine in veterinary medicine, a main compound of salicylanilide. Previous studies have reported that rafoxanide, as an inhibitor of BRAF V600E mutant protein, inhibits the growth of colorectal cancer, multiple myeloma, and skin cancer. However, its therapeutic effect on gastric cancer (GC) and the potential mechanism has not been investigated. Here, we have found that rafoxanide inhibited the proliferation of GC cells in vitro, arrested the cell cycle in the G0/G1 phase, and promoted apoptosis and autophagy in GC cells. Treatment with specific autophagy inhibitor 3-methyladenine drastically inhibited the apoptotic cell death effect by suppressing the switch from autophagy to apoptosis. Mechanistically, we found that rafoxanide inhibited the growth of GC cells in vitro by inhibiting the activity of the PI3K/Akt/mTOR signaling pathway. This process induced autophagy, which essentially resulted in the apoptosis of GC cells. Results from subcutaneous implanted tumor models in nude mice also indicated that rafoxanide inhibited the growth of GC cells in vivo. Taken together, our findings revealed that rafoxanide inhibited the growth of GC cells both in vitro and vivo, indicating a potential drug candidate for the treatment of GC.


Fine specificity of a monoclonal anti-testicular cell antibody for glycolipids with terminal N-acetyl-D-glucosamine structure.
The specificity of a mouse anti-testicular cell monoclonal antibody, J1, was investigated. Previous studies suggested that N-acetyl-D-glucosamine (GlcNAc) was a constituent of the determinant recognized by J1. When the antibody was tested against a variety of purified glycolipids containing this saccharide in terminal, penultimate or internal positions, J1 reacted only with species expressing terminal GlcNAc. The influence of oligosaccharide chain length, branch substitution, and haptenic valence on J1 binding was examined using glycolipids prepared by a weak acid hydrolysis and exoglycosidase digestion of bovine I-active ganglioside. Degree of binding was inversely proportional to chain length and was proportional to hapten valence. Failure of J1 to bind partially deglycosylated transferrin implied binding preference for GlcNAc beta 1----3Gal over GlcNAc beta 1----2Man. Immunofluorescence analysis of J1 binding to human neutrophils failed to detect lactotriosylceramide on their surface, although this glycolipid has previously been isolated from these cells, suggesting that this structure exists in a cryptic or intracellular form. Binding results were consistent with J1 having low affinity for GlcNAc or GlcNAc beta 1----3Gal on a variety of lacto-series glycolipids.


Synergistic and additive properties of the beta-globin locus control region (LCR) revealed by 5'HS3 deletion mutations: implication for LCR chromatin architecture.
Deletion of the 234-bp core element of the DNase I hypersensitive site 3 (5'HS3) of the locus control region (LCR) in the context of a human beta-globin locus yeast artificial chromosome (beta-YAC) results in profound effects on globin gene expression in transgenic mice. In contrast, deletion of a 2.3-kb 5'HS3 region, which includes the 234-bp core sequence, has a much milder phenotype. Here we report the effects of these deletions on chromatin structure in the beta-globin locus of adult erythroblasts. The 234-bp 5'HS3 deletion abolished histone acetylation throughout the beta-globin locus; recruitment of RNA polymerase II (pol II) to the LCR and beta-globin gene promoter was reduced to a basal level; and formation of all the 5' DNase I hypersensitive sites of the LCR was disrupted. The 2.3-kb 5'HS3 deletion mildly reduced the level of histone acetylation but did not change the profile across the whole locus; the 5' DNase I hypersensitive sites of the LCR were formed, but to a lesser extent; and recruitment of pol II was reduced, but only marginally. These data support the hypothesis that the LCR forms a specific chromatin structure and acts as a single entity. Based on these results we elaborate on a model of LCR chromatin architecture which accommodates the distinct phenotypes of the 5'HS3 and HS3 core deletions.


De-escalation of surgery for occult breast cancer with axillary metastasis.
Occult breast cancer presenting as axillary metastasis is rare and remains a diagnostic and therapeutic challenge. Evidence to guide clinical management is limited, and locoregional treatment remains nonstandardized and highly varied nationally. Historically, occult breast cancer was managed with modified radical mastectomy ± radiotherapy; however, equivalent local control and survival are observed with breast preservation and adjuvant whole breast radiotherapy. Axillary lymph node dissection remains the standard surgical approach to the axilla for occult breast cancer patients. De-escalating axillary surgery in a subset of occult breast cancer patients treated with neoadjuvant chemotherapy with good response to treatment may be appropriate, similar to the management of clinically node-positive patients in a known primary setting. As in other clinically node-positive breast cancer cases, thoughtful integration and tailoring of axillary surgery and regional nodal radiotherapy (for the varying extent of nodal burden) is an area of continued controversy and active investigation.


Characterization of antibody responses to purified HIV-1 gp120 glycoproteins fused with the molecular adjuvant C3d.
The HIV-1 exterior envelope glycoprotein gp120 binds receptor (CD4) and co-receptors (CCR5/CXCR4) and is a major target for neutralizing antibodies. The two functionally conserved regions of gp120 involved in receptor binding are conformational in nature. It is likely that the elicitation of neutralizing antibodies to these targets will benefit by presentation of these sites to the humoral immune system under physiologic conditions. Initially, we investigated the ability of the molecular adjuvant C3d to enhance antibody responses to variant gp120 glycoproteins in phosphate-buffered saline (PBS). We utilized a gp120 variant glycoprotein deleted of N- and C-terminal sequences (gp120DeltaC1/C5) originally designed to eliminate immunodominant, non-neutralizing epitopes and characterized this protein when fused to two C3d elements (gp120DeltaC1/C5(C3d)(2)). In PBS, the gp120DeltaC1/C5(C3d)(2) proteins are able to elicit gp120 binding antibodies more efficiently than gp120 lacking C3d moieties. We then asked if we could observe C3d-enhanced immunogenicity of gp120 in the presence of the classical oil-in-water adjuvant, Ribi. In the presence of the Ribi, which contains the TLR-4 agonist monophospholipid A (MPL), antibodies elicited by the gp120DeltaC1/C5(C3d)(2) were of higher titer than those elicited by the identical protein in PBS. To determine if the elicited secondary response was due to a synergy between the C3d repeats and the Ribi, we then inoculated gp120DeltaC1/C5 protein in Ribi and observed that similar titers of anti-gp120 antibodies were elicited in comparison to the gp120DeltaC1/C5(C3d)(2) protein also inoculated in Ribi adjuvant. In Ribi, there was a small but consistent increase in gp120-specific antibody titer of a gp120DeltaC1/C5(C3d)(2) prime followed by two gp120DeltaC1/C5 boosts compared to three inoculations of either the gp120DeltaC1/C5 proteins or the gp120DeltaC1/C5(C3d)(2) proteins alone. We conclude that the molecular adjuvant C3d demonstrates utility in conditions where physiologic presentation of native protein structures is desired, but may have less benefit in the context of a relatively potent protein adjuvant such as Ribi.


Differential distribution of alternatively spliced somatostatin receptor 2 isoforms (sst2A and sst2B) in rat spinal cord.
We have recently shown that the cytoplasmic tail of the somatostatin receptor sst2 undergoes alternative splicing giving rise to two isoforms, SSt2A and sst2B. In the present study, we have raised polyclonal antibodies that specifically detect either sst2A or sst2B and used these antisera for immunocytochemical localization of the receptor proteins in the rat spinal cord. sst2A-immunoreactivity formed a dense network consisting of neuronal perikarya and dendrites in the superficial layers of the dorsal horn. In contrast, prominent sst2B-immunoreactivity was found on neuronal perikarya and proximal dendrites throughout the gray matter of the spinal cord. Taken together, we show that alternative carboxy-terminal splicing is involved in cell-specific expression of somatostatin receptor sst2 isoforms in rat spinal cord, and that sst2A and sst2B mediate effects of somatostatin at different cellular sites.


Single versus double arterial anastomoses in combined second- and third-toe transplantation.
Combined second and third toe transplantation is one good option for reconstruction of multiple digit amputation. However, the use of one or two arteries for pedicle anastomoses, which may influence the vascular complication and success rate, has never been addressed in the literature. This study includes a retrospective review of 57 combined second and third toe transplantation in 54 patients performed from February of 1983 through December of 1996. Group I, composed of 41 transplantations, underwent one arterial anastomosis, and group II, composed of 16 transplantations, underwent double arterial anastomoses during surgery if there were two recipient arteries available or whenever the second and third toes showed inadequate blood perfusion after one arterial anastomosis. In group I, 10 transplantations (24.4 percent) required re-exploration with a success rate of 92.7 percent (38 out of 41 transplantations). In group II, only one transplantation (6.2 percent) required re-exploration with successful flap salvage. The success rate was 100 percent for group II. Because the re-exploration and success rates between groups I and II were not statistically significant according to two-tailed Fisher's exact test, the combined second and third toe transplantation is a reliable procedure using either single or double arterial anastomoses.


Statistical learning and spelling: Evidence from Brazilian prephonological spellers.
We analyzed the spelling attempts of Brazilian children (age 3 years, 3 months to 6 years, 0 months) who were prephonological spellers, in that they wrote using letters that did not reflect the phonemes in the words they were asked to spell. We tested the hypothesis that children use their statistical-learning skills to learn about the appearance of writing and that older prephonological spellers, who have had on average more exposure to writing, produce more wordlike spellings than younger prephonological spellers. We found that older prephonological spellers produced longer spellings and were more likely to use letters and digrams in proportion to their frequency of occurrence in Portuguese. There were also some age-related differences in children's tendency to use letters from their own names when writing other words. The results extend previous findings with learners of English to children who are learning a more transparent orthography.


Adamantyl-ureas with pyrazoles substituted by fluoroalkanes as soluble epoxide hydrolase inhibitors.
A series of soluble epoxide hydrolase (sEH) inhibitors containing halogenated pyrazoles was developed. Inhibition potency of the obtained compounds ranges from 0.8 to 27.5 nM. 1-Adamantyl-3-[(4,5-dichloro-1-methyl-1Н-pyrazol-3-yl)methyl]urea (3f, IC50 = 0.8 nM) and 1-[(Adamantan-1-yl)methyl]-3-[(4,5-dichloro-1-methyl-1Н-pyrazol-3-yl)methyl]urea (4f, IC50 = 1.2 nM) were found to be the most potent sEH inhibitors within the described series.