Novel carrier-mediated membrane-assisted three-phase liquid-liquid extraction coupled with liquid chromatography-mass spectrometry for the determination of eight biogenic amines in foods.
Analysis of biogenic amines (BAs) is of great importance due to their toxicity and usage as indicators of food freshness. In this study, membrane-assisted three-phase liquid-liquid extraction (MA-3pLLE) was proposed to integrate extraction and back-extraction into a single step using a specially designed U-shaped device. Parameters affecting the performance of the extraction method were optimized. Coupled with liquid chromatography-mass spectrometry, the method was successfully applied for the simultaneous determination of eight BAs without derivatization in apple juice, orange juice, red wine, soy sauce and milk granules, with satisfactory recoveries and RSDs. The calibration curve of the method was linear in the range of 1.00~100.00 ng/mL, with a correlation coefficient (r2) ≥ 0.9908. The limits of detection were in the range of 0.03~1.00 ng/mL. MA-3pLLE is efficient, reproducible, and green and has great potential for application in the one-step extraction of analytes from complex matrices.


The role of auditory feedback in the motor learning of music in experienced and novice performers.
Musical learning is related to the development of audio-visuomotor associations linking gestures with musical sounds. To study the role of auditory feedback in learning, 115 students (56 guitarists, 59 pianists) at the beginner, intermediate and advanced levels were recruited. Playing with sound (audio-motor feedback), mute practice (motor feedback), and piece listening (auditory feedback) were compared to first sight reading to assess the role of auditory and motor feedback in procedural learning. The procedure consisted of the execution of a standard piece for determining the students' level and 4 further music executions (every week for 4 weeks), preceded by different practice conditions (for 12 min, once a day, for 5 days). Real musical pieces (e.g., Segovia, Schubert, Bartók) were used. Performance evaluation focused on four macro-categories: note, rhythm, dynamics and smoothness. For both instruments, first-sight reading (A - M -) was associated with the worst performance: silent motor practice (A - M +) resulted in learning the rhythmic structure of the piece and in a smoother performance. Listening to pieces (A + M -) resulted in learning the agogics and in improving articulation and smoothness. Listening during performance (A + M +) resulted in fewer intonation errors. Interestingly, auditory feedback was more relevant for beginners than for advanced students, as evidenced by the greater benefits of listening during practice.


Sesbania rostrata root and stem nodule leghemoglobins: purification, and relationships among the seven major components.
By anion-exchange chromatography, the nitrogen fixing photosynthetic stem nodules and nonphotosynthetic root nodules of Sesbania rostrata are shown to contain the same seven major components of leghemoglobin (Lb), numbered LbI-LbVII in order of elution, although in different proportions. No novel component was found in photosynthetic nodules. All components of Sesbania Lb are monomeric, with molecular weights varying between 15,000 and 17,000, and at least six of them are separate gene products. It is suspected that variable conjugation with nonprotein moieties might be partially responsible for the molecular weight differences and anomalous behavior observed between isoelectric focusing and anion-exchange chromatography.


The significance of amelogenin loci from toothpicks as forensic evidence for sex determination.
DNA analysis has become the gold standard of all identification methods in forensic science. There are several primary and secondary sources of DNA samples in the field of forensics. Primary sources can be obtained directly from individuals, whereas secondary sources of DNA samples include items used by someone such as clothes, toothbrushes, and toothpicks. In Asian countries such as Korea, Japan, and Indonesia, toothpicks are often used, as the tip of toothpicks used to clear food particles in the interdental regions can be utilized to capture DNA samples from saliva. The use of saliva samples for DNA analysis in sex identification is based on short tandem repeats with the amelogenin (AMG) locus. The purpose of this study was to examine the concentration and purity of DNA on toothpicks as evidence in criminal case investigations, as well as its potential as sex determination material. Eight subjects were instructed to clean the interproximal side of their posterior teeth using a toothpick. Each toothpick sample was kept for 0 days (as a control), 14 days, and 20 days. The purity and DNA concentration of each sample were determined through DNA examination. After determining the concentration and purity of DNA from each sample, electrophoresis of the AMG loci was performed for sex determination. This study showed that the average concentration of DNA on toothpicks ranged from 425.25 to 796.25 μg/ml, and the average purity of DNA ranged from 1.09 to 1.13 μg/ml. The AMG gene produces 112 and 116 base pair amplicons from the X and Y chromosomes. Sex determination using DNA can be done using AMG loci, a protein found on the sex chromosomes (X and Y). The value of DNA concentration on toothpicks could be used to support forensic identification after 20 days at room temperature.


Memantine Mitigates Oligodendrocyte Damage after Repetitive Mild Traumatic Brain Injury.
Repetitive mild traumatic brain injury (rmTBI; e.g., sports concussions) is common and results in significant cognitive impairment, white matter injury and increased risk of neurodegeneration. Targeted therapies for rmTBI are lacking, though evidence from other injury models indicates that targeting N-methyl-d-aspartate (NMDA) receptor (NMDAR)-mediated glutamatergic toxicity might mitigate rmTBI-induced injury. We have previously shown that the NMDAR antagonist memantine lessens axonal injury and restores long term potentiation after rmTBI. Here, we evaluated whether the protective effects of memantine include oligodendrocyte specific mechanisms, as prior studies suggest that oligodendrocytes are particularly vulnerable to glutamatergic toxicity. Mice were subjected to rmTBI injury (5 injuries in 5 days) and randomized to treatment with memantine or with vehicle (n = 32/group). At the molecular level, oligodendrocyte counts and function (myelin basic protein, MBP) were assessed by immunohistochemistry and western blot at days 3, 7 and 28 days after the last injury. Axon integrity was assessed by neurofilament light chain (NF-l) expression and axonal ultrastructure was evaluated by electron microscopy. Compared to vehicle-treated mice, memantine-treated mice were protected against oligodendrocyte loss and decreased MBP expression at subacute time points after injury. Memantine treatment also protected against axon damage assessed by NF-l expression. These data suggest that the therapeutic effects of post-concussive NMDAR antagonism may in part work through oligodendrocyte specific mechanisms, which may have implications for long term neurodegenerative sequelae after multiple concussions.


Synchronization of activity rhythms with the tide in a saltmarsh collembolan Anurida maritima.
On a marine saltmarsh, the intertidal collembolan Anurida maritima (Guérin) has a well-defined tidal rhythm of activity on the mud surface between submerging tides: a large proportion of the individuals emerge from their underground refuges shortly after the retreat of the tide, forage there for a few hours, and retire underground at least 1 h before the return of the tide. This activity rhythm has a period of about 12.4 h and it persists in the field during sequences of non-submerging tides and in the laboratory away from the tides in constant light. In the field the level of activity is greatly reduced during the night. It is suggested that the function of this tidal rhythm is to enable individual Anurida to anticipate the return of the tide both during periods of regular, twice-daily tidal submergence and after a period of days when the habitat has not been covered by the tide.


Sulfur determination using the SiS diatomic molecule via HR-CS GF MAS and direct analysis of solid samples: A versatile method for different matrices.
In this work, a simple and reliable method was proposed for sulfur determination in different matrices using the diatomic molecule SiS via high-resolution continuum source graphite furnace molecular absorption spectrometry (HR-CS GF MAS) and direct analysis of solid samples. This investigation was carried out using six sulfur standard solutions, Na2S, Na2SO4, BeSO4, thiourea, l-cysteine and sulfamic acid, and nine certified reference materials (CRMs). All measurements were performed using the SiS analytical line at 282.910 nm with 400 μg Zr as permanent modifier and 20 μg Si in basic media as molecule forming reagent. The optimized pyrolysis and vaporization temperatures were 1200 and 2000 °C, respectively. The investigated sulfur sources presented similar analytical signals and statistically equal Aint values for the SiS molecule. Calibration curves with aqueous sulfur standard solutions were used to achieve the limits of detection (LOD) and quantification (LOQ) and the characteristic mass (m0) of 8.8, 29 ng mg-1 and 9.8 ng, respectively, and to determine sulfur in the CRMs. Considering that the investigated wavelength range contained multiple SiS lines, the LOD (2.5 ng mg-1), LOQ (8.4 ng mg-1), m0 (1.0 ng) and the working range (0.008-2.5 μg) were improved by using ten SiS lines (30 pixels) for the measurements. Despite differences in the CRM matrices, the SiS molecule was successfully employed to determine their sulfur concentrations, which were in agreement with the certified values at a confidence level of 95% through Student or Welch t-tests. Therefore, a simple, versatile and reliable method using the SiS molecule was developed to perform sulfur determination in a great variety of sample matrices via HR-CS GF MAS and direct analysis of solid samples.


The Impact of Bariatric Surgery on Diverticulitis Outcomes and Risk of Recurrent Hospitalizations in Adults with Clinically Severe Obesity.
Clinically severe obesity (SO) is a known risk factor for worsened outcomes and recurrence of acute diverticulitis. Paucity of data exist on outcomes of diverticulitis after bariatric surgery. The Nationwide Readmissions Database was queried for diverticulitis hospitalizations between the years 2010 and 2014. We restricted analysis to patients with SO and those who had bariatric surgery (BRS). Outcomes of mortality, surgical events, and recurrent diverticulitis admissions were compared using multivariable analysis and one-to-one propensity score matching. Among 52,274 diverticulitis admissions, 91.2% (47,694) patients had SO and 8.8% (4580) had prior BRS. Patients with SO had higher odds of suffering mortality on index diverticulitis admission when compared to those with prior BRS [adjusted odds ratio (aOR): 10.55; 95%CI 1.45,76.75]. Patients with SO were also more likely to undergo emergency surgery (aOR: 1.71; 95%CI 1.25,2.34) and colectomy (aOR: 1.45; 95%CI 1.26,1.68). Rates of recurrent diverticulitis readmissions within 30 days and 6 months were also higher in patients with SO compared to BRS patients (aOR: 7.94; 95%CI 1.09,57.83 and aOR: 1.98; 95%CI 1.14,3.43, respectively). Propensity score matching confirmed our findings of increased rates of mortality (OR: 17.28; 95%CI 2.02,147.6), recurrent diverticulitis, and worsened surgical outcomes within 30 days in patients with SO compared to BRS. This study is first to show improved outcomes and less recurrent hospitalizations for diverticulitis after bariatric surgery compared to patients with clinically severe obesity. Further studies are needed to understand mechanisms leading to this improvement and the role of weight loss in prevention of severe diverticulitis.


Use of lymph node ratio to guide clinical decision-making concerning adjuvant radiotherapy in pT1-2N1 rectal cancer.
Lymph node metastases are uncommon in pT1-2 rectal cancer. pT1-2N1 are often characterized with low tumor burden and intermediate prognosis. Therefore, adjuvant radiotherapy (ART) is controversial in these patients. This study aimed to investigate the value of ART in pT1-2 rectal cancer and evaluate the guiding role of lymph node ratio (LNR) for utilization of ART. pT1-2N1 rectal cancer patients who received surgery without neoadjuvant radiotherapy between 2000 and 2018 with at least 12 lymph node harvest were extracted from the Surveillance, Epidemiology and End Results (SEER) database. We used time-dependent receiver operating characteristic (ROC) analysis to determine the optimal cutoff of LNR. Kaplan-Meier methods and Cox proportional hazards regression models were performed to determine the prognostic value of ART in pT1-2N1 rectal cancer patients and subgroups stratified by LNR. A total of 674 and 1321 patients with pT1N1 and pT2N1 rectal cancer were eligible for analysis. There was no statistical cancer-specific survival (CSS) difference in pT1N1 rectal cancer patients between receiving and not receiving ART (P = 0.464). The 5-year CSS was 89.6% and 83.2% in pT2N1 rectal cancer patients between receiving and not receiving ART, respectively (P = 0.003). A total of 7.0% was identified as the optimal cutoff value of LNR. Survival improvement offered by ART was only found in LNR ≥ 7.0% subgroup (5-year CSS: 89.5% versus 79.6%, P = 0.003) instead of LNR < 7.0% subgroup (5-year CSS: 89.9% versus 86.3%, P = 0.208). ART show substantial survival benefit in pT2N1 rectal cancer patients with LNR ≥ 7.0%, warranting the conventional adoption of ART in this subgroup.


Neuropsychological instruments for bipolar disorders: A systematic review on psychometric properties.
Cognitive deficits are a core feature of bipolar disorder (BD) that persist during the euthymic phase and affect global functioning. However, nowadays, there is no consensus on the optimal tool to capture cognitive deficits in BD. Therefore, this review aims to examine the psychometric properties of tools commonly used to assess cognitive functioning in BD. Literature search was conducted on PubMed and Web of Science databases on August 1, 2022 and on April 20, 2023, yielding 1758 de-duplicated records. Thirteen studies fulfilled the inclusion criteria and were included in the review. All tools examined showed acceptable-to-good psychometric properties suggesting that both brief cognitive screeners and comprehensive batteries may be appropriate for detecting or monitoring cognitive changes in BD. Methodological differences between the included studies precluded a direct comparison of the results. Further research is needed to investigate the psychometric properties of cognitive tools that assess also affective and social cognition. The tools examined appear sensitive enough to distinguish between BD patients with versus without cognitive deficits, however, an optimal tool has not yet been identified. The applicability and clinical utility of the tools may depend on multiple factors such as available resources. That said, web-based instruments are expected to become the first-choice instrument for cognitive screening as they can be applied on a large scale and at an affordable cost. As for second-level assessment instruments, the BACA shows robust psychometric properties and tests both affective and non-affective cognition.


Weak hierarchies associated with similarity measures--an additive clustering technique.
A new and apparently rather useful and natural concept in cluster analysis is studied: given a similarity measure on a set of objects, a sub-set is regarded as a cluster if any two objects a, b inside this sub-set have greater similarity than any third object outside has to at least one of a, b. These clusters then form a closure system which can be described as a hypergraph without triangles. Conversely, given such a system, one may attach some weight to each cluster and then compose a similarity measure additively, by letting the similarity of a pair be the sum of weights of the clusters containing that particular pair. The original clusters can be reconstructed from the obtained similarity measure. This clustering model is thus located between the general additive clustering model of Shepard and Arabie (1979) and the standard hierarchical model. Potential applications include fitting dendrograms with few additional nonnested clusters and simultaneous representation of some families of multiple dendrograms (in particular, two-dendrogram solutions), as well as assisting the search for phylogenetic relationships by proposing a somewhat larger system of possibly relevant "family groups", from which an appropriate choice (based on additional insight or individual preferences) remains to be made.


Chemical-analytical characterization and leaching of heavy metals associated with nanoparticles and microplastics from commercial face masks and the abundance of personal protective equipment (PPE) waste in three metropolitan cities of South America.
In this study, we surveyed the presence of personal protective equipment (PPE) waste on the streets of Bogotá-Colombia, Lima-Perú, and Mar del Plata-Argentina. Furthermore, this work is also focused on the release capacity of Ag, Cu, and Zn metals associated with nanoparticles, and microplastics (MPs) from textile face masks (TFMs) and disposable face masks. According to our results, an association between low-income areas and PPE waste was found, which may be related to the periodicity of waste collection and economic activity. Polymers, like polypropylene, cotton-polyester, and additives, such as CaCO3, MgO, and Ag/Cu as nanoparticles, were identified. TFMs released high levels of Cu (35,900-60,200 μg·L-1), Zn (2340-2380 μg·L-1), and MPs (4528-10,640 particles/piece). Metals associated with nanoparticles leached by face masks did not present any antimicrobial activity against P. aeruginosa. Our study suggests that TFMs may leach large amounts of polluting nano/micromaterials in aquatic environments with potential toxicological effects on organisms.


Development and validation of a sensitive assay for analysis of midazolam, free and conjugated 1-hydroxymidazolam and 4-hydroxymidazolam in pediatric plasma: Application to Pediatric Pharmacokinetic Study.
Pharmacokinetic, pharmacodynamic and pharmacogenomic studies of midazolam are currently being performed in critically ill children to find suitable dose regimens. Sensitive assays using small volumes of plasma are necessary to determine the concentrations of midazolam and its respective metabolites in pediatric studies. Midazolam is metabolized to hydroxylated midazolam isomers, which are present as free as well as the corresponding glucuronide conjugates. A high-performance liquid chromatographic method with tandem mass spectrometry has been developed and validated for the quantification of midazolam, and free and total 1-hydroxymidazolam and 4-hydroxymidazolam metabolites in small volumes of plasma. Cleanup consisted of 96-well μ-elution solid phase extraction (SPE). The analytes were separated by gradient elution using a C18 analytical column with a total run time of 5min. Multiple reaction monitoring was employed using precursor to product ion transitions of m/z 326.2→291.3 for midazolam, m/z 342.1→203.0 for 1-hydroxymidazolam, m/z 342.1→325.1 for 4-hydroxymidazolam and m/z 330.2→295.3 for 2H4-midazolam (internal standard). Since authentic hydroxymidazolamglucuronide standards are not available, samples were hydrolyzed with β-glucuronidase under optimized conditions. Assay conditions were modified and optimized to provide appropriate recovery and stability because 4-hydroxymidazolam was very acid sensitive. Standard curves were linear from 0.5 to 1000ng/mL for all three analytes. Intra- and inter day accuracy and precision for quality control samples (2, 20, 200 and 800ng/mL) were within 85-115% and 15% (coefficient of variation), respectively. Stability in plasma and extracts were sufficient under assay conditions. Plasma samples were processed and analyzed for midazolam, and free 1-hydroxymidazolam and 4-hydroxymidazolam metabolites. Plasma samples that were hydrolyzed with β-glucuronidase were processed and analyzed for midazolam, and total 1-hydroxymidazolam and 4-hydroxymidazolam metabolites under the same assay conditions. The difference in concentration between the total and free hydroxymidazolam metabolites provided an estimate of conjugated hydroxymidazolam metabolites. The combination of 96-well μ-elution SPE and LC-MS/MS allows reliable quantification of midazolam and its metabolites in small volumes of plasma for pediatric patients. This assay is currently being successfully utilized for analysis of samples from ongoing clinical trials.


Evolution of IgM, IgE and IgG(1-4 )antibody responses in early childhood monitored with recombinant allergen components: implications for class switch mechanisms.
The formation of IgE antibodies against environmental allergens represents the hallmark of type I allergy. Data from in vitro cultured cells and experimental animal models provide controversial evidence for isotype switching from IgM to IgE production via sequential as well as non-sequential (i.e. direct) class switch. We analyzed the evolution of IgE responses in 11 children developing birch pollen and/or grass pollen allergy during the first 7 years of life using purified recombinant allergen molecules (major birch pollen allergen, Bet v 1; major timothy grass pollen allergens, Phl p 1, Phl p 2, Phl p 5). Demographic, clinical and serological data indicated a postnatal sensitization to pollen allergens. A parallel development of IgG(1-4) and IgE responses to recombinant allergen molecules compatible with a strictly sequential class switch to IgE was observed only in one child. The only partly synchronized and dissociated development of allergen-specific antibody responses found in all other cases can be best explained by a partly sequential class switch involving few switch stations or, more likely, by direct class switching. Kinetics and courses of allergen-specific antibody responses (IgM, IgG(1-4), IgE) during the first years of life suggest that, once established, allergen-specific IgE responses are driven by antigen contact rather than by cytokines controlling class switch to IgE.


Wide local excision, Mohs micrographic surgery, and reconstructive options for treatment of dermatofibrosarcoma protuberans of the breast: A retrospective case series from Mayo Clinic.
Dermatofibrosarcoma protuberans (DFSP) of the breast is a dermal fibroblastic neoplasm requiring wide excisional margins due to recurrence rates ranging from 26 to 60%. The current literature on reconstructive options and utility of Mohs micrographic surgery for DFSP of the breast is scarce. We describe surgical management of DFSP of the breast at our institution with the largest case series reported to date. A retrospective review was performed of women who underwent surgery for DFSP of the breast at our institution between 1990 and 2019. Continuous data was summarized using mean, median, and range; categorical data was summarized with frequency count and percentage. Preoperative lesion size and postoperative defect size were evaluated using 2-sided Fisher exact test, and p-values < 0.05 were considered statistically significant. Nine patients underwent wide local excision (WLE) with reconstruction including pedicled latissimus dorsi flaps (n = 2), local flap advancement (n = 2), mastectomy with implant (n = 1), oncoplastic breast reduction (n = 1), and skin grafts (n = 3). Nine underwent Mohs micrographic surgery (MMS) with complex primary closure. Mean postoperative maximum wound defect size for WLE was 10.8 cm versus 7.0 cm for MMS with no statistical significance (p = 0.77). Mean preoperative maximum lesion size for WLE was 6.4 cm versus 3.3 cm for MMS with no statistical significance (p = 0.07). Complications with WLE included wound dehiscence in three patients and seroma in one patient. No complications were reported with MMS and primary closure. Recurrence was reported in one WLE patient, which was successfully detected despite flap coverage and resected without complications. Median follow-up for the patients without recurrence was 5.0 years, with two patients in MMS cohort lost to follow-up. Five-year overall survival was 100%. MMS and WLE are both viable surgical options for managing DFSP of the breast. MMS could potentially minimize reconstructive needs due to smaller average defect size and result in fewer complications but may also result in asymmetry. Immediate flap reconstruction, especially in larger defects, can achieve excellent aesthetic outcomes for patients with DFSP of the breast without compromising detection of disease recurrence.


Influence of N and K fertilization and growth temperature on 13C/12C ratios of timothy (Phleum pratense L.).
Measurements have been reported of carbon isotope ratios of timothy grown at different temperatures and with varying nitrogen and potassium supplies. Both total plant tissue and extracted plant tissue have been analyzed. The 13C/12C ratios were found to vary both with temperature and with nutrient level; the highest values of δ13C were found under the most optimum growth conditions.


Ocular myopathy without ophthalmoplegia can be a form of mitochondrial myopathy.
To characterize muscle pathology in 3 cases affected by ocular myopathy with eyelid ptosis and upper facial weakness, but without ophthalmoplegia, light microscopy and ultrastructural study were performed on levator palpebrae, orbicularis oculi and deltoid muscle biopsies. While levator palpebrae proved uninformative because of the massive fibrous degeneration of muscle, orbicularis oculi biopsies showed histochemical and ultrastructural alterations indicating a mitochondrial involvement, resembling that reported in ocular mitochondrial myopathies (OMM). On the other hand very mild aspecific findings were observed in deltoid. We suggest that these cases with ocular myopathy and without ophthalmoplegia should be considered a partial or initial form of OMM.


In vivo evaluation of nicotine lyophilised nasal insert in sheep.
The nasal route offers an attractive means of delivering a drug directly to the systemic circulation and avoiding hepatic first-pass metabolism, although rapid mucociliary clearance can be detrimental to nasal absorption. The in vitro and in vivo characteristics of a nasal insert formulation prepared by lyophilisation of a viscous HPMC gel solution designed to overcome this problem were studied. In vitro release of nicotine from the lyophilised insert was compared with powder and spray formulations. Stability and characterisation studies were carried out using dynamic vapour sorption, scanning electron microscopy and HPLC analysis. Nicotine formulations were administered to eight wether sheep in a randomised four-way cross-over study, and plasma nicotine assessed comparing the nasal insert formulation with conventional nasal powder, nasal spray and IV doses. In vitro release studies demonstrated prolonged nicotine release from the nasal insert formulation compared to a powder and liquid. In vivo plasma profiles appeared to show prolonged plasma nicotine levels compared to the conventional formulations, although T(max), C(max) and AUC parameters for the insert were not significantly different due to high variability in the pharmacokinetic data. In conclusion, the nasal insert displayed a promising prolonged plasma profile, which must be investigated further to provide statistical significance to prove the effect.


The effect of a locally tailored intervention on the uptake of preconception care in the Netherlands: a stepped-wedge cluster randomized trial (APROPOS-II study).
The preconception period provides a window of opportunity for interventions aiming to reduce unhealthy lifestyle behaviours and their negative effect on pregnancy outcomes. This study aimed to assess the effectiveness of a locally tailored preconception care (PCC) intervention in a hybrid-II effectiveness implementation design. A stepped-wedge cluster randomized controlled trial was performed in four Dutch municipalities. The intervention contained a social marketing strategy aiming to improve the uptake (prospective parents) and the provision (healthcare providers) of PCC. Prospective parents participated by administering a questionnaire in early pregnancy recalling their preconceptional behaviours. Experiences of healthcare providers were also evaluated through questionnaires. The composite primary outcome was adherence to at least three out of four preconceptional lifestyle recommendations (early initiation of folic acid supplements, healthy nutrition, no smoking or alcohol use). Secondary outcomes were preconceptional lifestyle behaviour change, (online) reach of the intervention and improved knowledge among healthcare providers. A total of 850 women and 154 men participated in the control phase and 213 women and 39 men in the intervention phase. The composite primary outcome significantly improved among women participating in the municipality where the reach of the intervention was highest (Relative Risk (RR) 1.57 (95% Confidence Interval (CI) 1.11-2.22). Among women, vegetable intake had significantly improved in the intervention phase (RR 1.82 (95%CI 1.14-2.91)). The aimed online reach- and engagement rate of the intervention was achieved most of the time. Also, after the intervention, more healthcare providers were aware of PCC-risk factors (54.5% vs. 47.7%; p = 0.040) and more healthcare providers considered it easier to start a conversation about PCC (75.0% vs. 47.9%; p = 0.030). The intervention showed some tentative positive effects on lifestyle behaviours among prospective parents. Primarily on vegetable intake and the knowledge and competence of healthcare providers. The results of this study contribute to the evidence regarding successfully implementing PCC-interventions to optimize the health of prospective parents and future generations. Dutch Trial Register: NL7784 (Registered 06/06/2019).


A randomized trial of N-acetylcysteine for prevention of trimethoprim-sulfamethoxazole hypersensitivity reactions in Pneumocystis carinii pneumonia prophylaxis (CTN 057). Canadian HIV Trials Network 057 Study Group.
Hydroxylamine derivatives of sulfamethoxazole may be the reactive metabolites that cause adverse reactions to trimethoprim-sulfamethoxazole (TMP-SMX). The increased frequency of reactions observed in HIV-positive individuals is hypothesized to be due to systemic glutathione deficiency and a decreased ability to scavenge these metabolites. Two hundred and thirty-eight patients were randomized to receive or not receive N-acetylcysteine (3 g of the 20% liquid solution) 1 hour before each dose of TMP-SMX (trimethoprim 80 mg, sulfamethoxazole 400 mg) twice daily, which was initiated as primary Pneumocystis carinii pneumonia prophylaxis. Forty-five patients had to discontinue TMP-SMX within 2 months because of fever, rash, or pruritus including 25 of 102 patients (25%) who were receiving TMP-SMX alone and 20 of 96 patients (21%) who were randomized to TMP-SMX and N-acetylcysteine. The difference between treatment groups is 4% (95% confidence interval [CI]: -16%, +9%). No independent association was found with the hypersensitivity reaction and age, gender, race, HIV risk factor, prior AIDS, concurrent use of fluconazole, or baseline CD4. N-acetylcysteine at a dose of 3 g twice daily could not be shown to prevent TMP-SMX hypersensitivity reactions in patients with HIV infection.


Prediction of early recurrence of hepatocellular carcinoma within the Milan criteria after radical resection.
Approximately 50% hepatocellular carcinoma patients meeting the Milan criteria utilized to develop an improved prognostic model for predicting the recurrence in these patients. Using univariate and multivariate analysis, cytokeratin-19 and glypican-3 expression patterns, tumor number and histological grading from eight putative prognostic factors comprised the risk factor scoring model to predict the tumor recurrence. In the training cohort, the area under roc curve (AUC) value of the model was 0.715 [95% confidence interval (CI) = 0.645-0.786, P<0.001], which was the highest among all the parameters. The performance of the model was assessed using an independent validation cohort, wherein the AUC value was 0.760 (95% CI=0.647-0.874, P<0.001), which was higher than the other factors. The results indicated that model had high performance with adequate discrimination ability. Moreover, it significantly improved the predictive capacity for the recurrence in patients with hepatocellular carcinoma within the Milan criteria after radical resection.


A randomized, double-blind multicentre clinical trial comparing the efficacy of calcium dobesilate with placebo in the treatment of chronic venous disease.
To assess the efficacy of calcium dobesilate on the quality-of-life (QoL) of patients with chronic venous disease (CVD). Randomised, parallel, double blind, placebo-controlled clinical trial. Patients were recruited from vascular surgery clinics and randomised to 500mg capsules of calcium dobesilate twice a day for 3 months or placebo. The primary outcome measure was 'QoL after 3 months' treatment measured by the specific Chronic Insufficiency Venous International Questionnaire (CIVIQ). Secondary outcomes were QoL at 12 months and assessment of the CVD signs and symptoms. The principal analysis was undertaken on the intention-to-treat (ITT) data. Five hundred and nine patients were recruited (246 to calcium dobesilate and 263 to placebo). The analysis of the 'QoL after 3 months' showed no significant differences between groups (p=0.07). For secondary outcomes, oedema and symptoms of CVD, there were no significant differences between groups. In a multi-factorial analysis, the 'QoL at 12 months' was better in the calcium dobesilate group than in placebo group (p=0.02). Treatment with calcium dobesilate was not found to be superior to placebo on the QoL of CVD patients. The sustained effect of calcium dobesilate observed after treatment should be confirmed in future studies.


The development of COMRADE--a patient-based outcome measure to evaluate the effectiveness of risk communication and treatment decision making in consultations.
Some instruments have been developed to evaluate 'decision effectiveness' such as the 'satisfaction with decision (SWD)' and 'decision conflict (DCS)' scales and are validated for the US context. Patients identify further outcome domains that are not fully covered in these scales. We developed a patient-based outcome measure to evaluate risk communication and decision making effectiveness, addressing these further domains and validated for use in the UK. Formulation of items was based on literature review and key informant interviews. The instrument was piloted and developed (including psychometric assessment) in the context of a trial of decision making and risk communication interventions in general practice. Patients were recruited to the trial with known atrial fibrillation, prostatism, menorrhagia or menopausal symptoms. High response rates (96% in first phase) indicated acceptability to patients. A 20 item patient-based outcome measure, with two sub-scales for 'risk communication' and 'confidence in decision', was produced. It allows for paternalistic, shared or informed choice decision making models. Usage in further studies will facilitate systematic reviews. Consequently, lessons for wider application of these interventions in practice, and policy implications regarding greater patient involvement should then be clearer.


Mixed connective tissue disease, Sjögren's syndrome, and abdominal pseudolymphoma.
A young female with mixed connective tissue disease (MCTD) presented with the unusual occurrence of abdominal pseudolymphoma. Although the pseudolymphoma could be related to concomitant sicca complex, some data suggest that MCTD, as such, could carry a high incidence of malignancy.


Interleukin-2 promotes survival and neurite extension of cultured neurons from fetal rat brain.
We investigated the effects of interleukin-2 (IL-2) on the survival and morphology of primary cultured neurons from fetal rat brain. Addition of recombinant human IL-2 significantly supported the survival of brain neurons in high cell density culture, but did not show any effect on the neuronal survival in low cell density culture. In addition, IL-2 significantly promoted the neurite elongation and branching of hippocampal neurons in low cell density culture. These results suggest that IL-2 supports neuronal survival indirectly and promotes neuritogenesis by directly acting on brain neurons.


Detection of DNA strand breaks in Escherichia coli treated with platinum(IV) antitumor compounds.
DNA strand breaks were observed in bacteria treated with Pt(IV) but not Pt(II) antitumor compounds by two methods. First, compounds which cause DNA strand breaks produced an SOS induction signal which was detected by a rapid bacterial assay. In addition, the capacity of these compounds to cut DNA in vivo was directly measured by agarose gel electrophoresis of pBR322 DNA extracted from bacteria treated with these drugs. cis-Diamminetetrachloroplatinum(IV) (cis-DTP) and cis-dichloro-trans-dihydroxo-cis-bis(isopropylamine)-platinum(IV) (iproplatin) produced strand breaks in both assays while cis-diamminedichloroplatinum(II) (cisplatin) did not. These results indicate that Pt(IV) antitumor complexes may cause DNA damage in vivo which is not produced by Pt(II) compounds.


Identification and Analysis of Essential Genes in Streptococcus mutans with Transposon Sequencing.
Transposon sequencing (Tn-seq) has greatly accelerated the rate at which gene function can be profiled in microbial organisms. This technique has been applied to the study of the dental caries pathogen Streptococcus mutans where it has been used to generate large transposon mutant libraries. Coupled with high-throughput sequencing and bioinformatics tools, culture of these transposon mutant libraries has facilitated the identification of essential and conditional essential genes. In this chapter, we describe a procedure for performing Tn-seq studies in S. mutans that covers pooled transposon mutant construction, in vitro culture, and DNA library sequencing and data analysis.


Jhe in Gryllus assimilis: cloning, sequence-activity associations and phylogeny.
The 458 amino acid sequence of a mature JHE protein from the cricket Gryllus assimilis was identified after isolating the partial cDNA sequence encoding this protein from a fat body and midgut cDNA library. This hemimetabolan JHE sequence shows over 40% amino acid similarity to the known JHE sequences of several holometabolous insects. It also includes previously determined peptide sequences for G. assimilis JHE as well as two other motifs associated with JHE enzymes in holometabolous insects. The predicted molecular weight of the protein agrees with that of the JHE previously purified from G. assimilis. Partial genomic sequence encoding the Jhe contains two large (1330 and 2918bp) introns. No coding DNA sequence variation was observed over a 1293bp region between selected lines differing six to eight-fold in hemolymph JHE activity. However, a 19bp indel was found in one of the introns; the insertion was strongly associated with elevated hemolymph activity, both in the selected lines and in the F(2) progeny of crosses between them. Phylogenetic analyses localised the G. assimilis JHE to a clade containing dipteran and coleopteran JHEs, with lepidopteran JHEs occurring in a separate clade.


Excitatory neurons in paraventricular hypothalamus contributed to the mechanism underlying acupuncture regulating the swallowing function.
Paraventricular hypothalamus (PVH) is demonstrated to regulate stress, feeding behaviors, and other related homeostatic processes. However, no direct evidence has been investigated for the role of PVH in swallowing function. Acupuncture therapy at Lianquan (CV23) acupoint has been reported to improve the swallowing function in clinical trials, but its underlying mechanism still needs to be uncovered. Thus, we aimed to explore whether PVH involved the acupuncture mediated regulating swallowing function. Chemogenetics, electromyography (EMG) recording, and immunofluorescence staining methods were combined to demonstrate that neurons in PVH could be activated by electroacupuncture (EA) stimulation at CV23, and this neuronal cluster was represented as excitatory neurons. Furthermore, we mapped both the inputs and outputs of PVH neurons using viral tracing. The neurons in PVH projected with the brain regions, including parabrachial nucleus (PBN) and the solitary tract nucleus (NTS), which both participated in the swallowing process. The EA function regulating the swallowing was attenuated after inhibiting the neurons in PVH in the post stroke dysphagia. In conclusion, this study suggested that EA at CV23 could regulate swallowing function involving the excitatory neurons in PVH.


Cholesterol metabolism in spontaneously hypertensive rats.
The absorption of radioactive cholesterol tended to be increased, and the catabolism of cholesterol detected by the decay of radioactive cholesterol in the serum after the intraperitoneal injection was delayed in ALR in comparison with normotensive WK rats. The reactive hypercholesterolemia in the ALR which had been selectively bred for a greater hypercholesterolemia was ascribed to the decreased cholesterol catabolism.


Efficacy of Colchicine in the Treatment of COVID-19 Patients: A Systematic Review and Meta-Analysis.
We assessed the efficacy of colchicine in COVID-19 patients through a systematic review. Six databases were searched until March 2022 for studies assessing colchicine versus control in hospitalized patients with COVID-19. The primary outcome was mortality, and secondary outcome was length of hospitalization. Inverse variance and random effect meta-analyses were performed. The strength of evidence was assessed using GRADE. Nine studies (five randomized clinical trials (RCTs) and four non-randomized studies of intervention (NRSI); n = 13,478). Colchicine did not reduce mortality in comparison with the standard of care in RCTs (RR 0.99; 95%CI 0.90 to 1.10; p = 0.90); however, it did reduce mortality in NRSI studies (RR 0.45; 95%CI 0.26 to 0.77; p = 0.02). In the analysis of RCTs, colchicine did not reduce the length of hospitalization in comparison with the standard of care (MD: -2.25 days; 95%CI: -9.34 to 4.84; p = 0.15). Most studies were scored as having a high risk of bias. Quality of evidence was very low for primary and secondary outcomes. Colchicine did not reduce the mortality and length of hospitalization in comparison with the standard of care in hospitalized patients with COVID-19. The published evidence is insufficient and of very low quality to recommend treatment in patients with COVID-19.


Retracted: Bacteria diversity overview and endoglucanase assessment from Himalayan Tapovan geothermal spring.
The above article from the Journal of Basic Microbiology, published online on 25 August 2015 in Wiley Online Library as Early View (http://onlinelibrary.wiley.com/doi/10.1002/jobm.201500135/pdf), has been retracted by agreement between Naveen Kumar Navani and Ranjana Pathania, the Editor-in-Chief and Wiley-VCH GmbH & Co. KGaA. The retraction has been agreed because the article has been submitted and approved for publication by Jitendra Kumar Sahoo without consent in any form by the named co-authors Naveen Kumar Navani and Ranjana Pathania.


Use of clinical practice guidelines on long-term prophylaxis in severe hemophilia in France: a retrospective audit.
To evaluate the impact of the 2002 guidelines on the current status of prophylaxis in French children with severe hemophilia A or B. Clinical information was captured, in a prospective way, using FranceCoag Network. We retrospectively studied 291 patients with severe (<1 IU/dL) hemophilia A and B, with no history of inhibitors. Our results demonstrate that the availability of national medical guidelines has improved clinical practice in France. In the past decade, the proportion of children with severe hemophilia undergoing prophylaxis has shown a significant 2- to 3-fold increase: ∼80% of these children>3 years of age are now receiving prophylaxis. In severe hemophilia A and B, the age at which prophylaxis commences has significantly decreased: 4.0 and 6.1 years for the period 1996-1999 as opposed to 1.8 and 1.4 years for the period 2004-2007 (P=.0001). Long-term clinical and physical evaluations of patients will be necessary to establish the benefits of this increase in prophylactic treatment on the prevention of hemophilic arthropathy.


Racial disparity, primary care, and specialty referral.
The study examines the role of primary care physicians (PCP) in reducing racial disparities in referral-sensitive admissions. The study examined hospital discharges of New York residents in the age group 20 to 64 hospitalized either in New York or in any of three contiguous states -- New Jersey, Pennsylvania, or Connecticut -- using complete discharge files for the four states in 1995. The discharge data were linked to the Area Resource File and the American Hospital Association's survey files for 1995. The study used multivariate logistic models to compare the effect of PCP supply on referral-sensitive (discretionary) admissions versus marker admissions (urgent, in sensitive to primary care) or whites, blacks, and Hispanics. Compared with marker admissions, an increased PCP density in an area was associated with a higher probability of black admissions than white admissions for referral-sensitive procedures. Our analysis suggests that the supply of primary care may be important for admissions for referral-sensitive procedures. Nonwhites, especially blacks, may have greater access to these procedures through increased PCP supply. Increased PCP density may significantly narrow the racial disparity in specialty referrals and improve the referral process for these procedures for nonwhites.


Political interpretation of scientific evidence--case study of breast cancer screening policies around the world.
Early detection of breast cancer has been studied for effectiveness in reducing mortality for more than four decades. During this period numerous studies took place, with more than half a million participating women. In spite of large amounts of nonconflicting data, different countries took varying lengths of time to establish a policy, and came up with a variety of policies, quite different from one another. Inherent differences between the countries in structure of the health system, in the commitment to public health activities, and in opinions and health habits of the relevant populations may explain these different outcomes.


Predictors of change in participation rates following acquired brain injury: results of a longitudinal study.
The purpose of this study was (1) to examine the changes in participation rates over 1 year among children and adolescents after acquired brain injury and (2) to explore the effect of child and family factors on these changes. The participation levels of 136 children and young people (88 males; 48 females; age range 4y 11mo-17y 6mo; mean age 11y 6mo) after acquired brain injury (3≤ Glasgow Coma Scale score ≤15; mean 12.8) were assessed three times: at their return to school, and at 8 and 12 months after returning to school. The Children's Assessment of Participation and Enjoyment measured the participants' diversity and intensity of participation in out-of-school activities. At baseline, information on general family functioning and medical and demographic information was collected as possible predictors. Mixed-effect model analyses of participation scores were performed while controlling for child's age at injury. The severity of the injury explained rates of change across time for participation intensity in recreational, physical, and social activities. Household income influenced changes in the intensity of recreational activities, whereas family functioning predicted changes in the diversity of skill-based activities. Participation is a relevant outcome of recovery that needs to be assessed and monitored post brain injury. Special attention can be directed to severity of injury and family functioning when developing intervention plans.


The ketone body β-hydroxybutyric acid influences agouti-related peptide expression via AMP-activated protein kinase in hypothalamic GT1-7 cells.
β-Hydroxybutyric acid (BHBA) acts in the brain to influence feeding behaviour, but the underlying molecular mechanisms are unclear. GT1-7 hypothalamic cells expressing orexigenic agouti-related peptide (AGRP) were used to study the AMP-activated protein kinase (AMPK) pathway known to integrate dietary and hormonal signals for food intake regulation. In a 25 mM glucose culture medium, BHBA increased intracellular calcium concentrations and the expression of monocarboxylate transporter 1 (MCT1 (SLC16A1)). Phosphorylation of AMPK-α (PRKAA1 and PRKAA2) at Thr(172) was diminished after 2 h but increased after 4 h. Its downstream target, the mammalian target of rapamycin, was increasingly phosphorylated on Ser(2448) after 2 h but not changed after 4 h of BHBA treatment. After 4 h, BHBA treatment also increased Agrp mRNA expression. This increase was prevented by preincubation with the AMPK inhibitor Compound C. The inhibition of MCT1 activity by p-hydroxymercuribenzoate suppressed BHBA-stimulated AMPK phosphorylation but did not prevent BHBA-induced Agrp mRNA expression. This finding demonstrates that BHBA triggers the AMPK pathway resulting in orexigenic signalling under 25 mM glucose culture conditions. Under conditions of 5.5 mM glucose, however, BHBA marginally increased intracellular calcium but significantly decreased AMPK phosphorylation and Agrp mRNA expression, demonstrating that under physiological conditions BHBA reduces central orexigenic signalling.


Factors affecting duration of post-surgical orthodontics in the Surgery First/Early Approach: A retrospective study.
Surgery First Approach (SFA) and Surgery Early (SE) are considered promising alternatives, compared to the conventional three-stages orthodontic-surgical approach, for treatment of dento-maxillofacial deformities. However, many features need further study, like the role of the orthodontist. Aim of the study was to analyse the clinical characteristics of patients who underwent SFA and SE, and if differences in duration of orthodontics could be influenced by clinical features. A retrospective research was performed on patients who met the inclusion criteria for SFA (31) and SE (12), of the total of patients affected by dento-facial deformities in our Unit (191) in the period 2012-2017. After collection of clinical data, duration of orthodontics, age, pre-treatment PAR Index, ANB angle, amount of the curve of Spee were compared. A regression analysis evaluated if these clinical parameters, together with type of bracket and type of intervention, could influence the duration of post-surgical orthodontics. All patients who performed the SFA/SE were affected by class III, II and active Unilateral Condylar Hyperplasia (22% of total population). Pre-treatment mean differences of age (p = 0.0518), PAR Index (p = 0.0916), curve of Spee (p = 0.1006) between groups were not statistically significant. A statically significant difference was found for the overall duration of therapy, for the significant shorter duration of pre-surgical orthodontics, while the difference of post-surgical orthodontics duration was not significant (p = 0.4753). Type of bracket (rho=-0.19039, p = 0.266) and intervention performed (rho=-0.11522, p = 0.5034) were not correlated with duration of post-surgical orthodontics, as well as pre-treatment PAR Index, ANB angle and depth of the curve of Spee. Surgery First/Early Approach is a therapeutic choice that could be performed only in patients affected by specific malocclusions and who exactly meet indications. Protocol and post-surgical occlusal stability are factors that should influence the duration of therapy more than clinical characteristics.


The sensory-efferent function of capsaicin-sensitive sensory neurons.
Capsaicin-sensitive sensory neurons convey to the central nervous system signals (chemical and physical) arising from viscera and the skin which activate a variety of visceromotor and neuroendocrine reflexes integrated at various levels (intramurally in peripheral organs, at level of prevertebral ganglia, spinal and supraspinal level). Much evidence is now available that peripheral terminals of certain sensory neurons, widely distributed in skin and viscera have the ability to release, upon adequate stimulation, their transmitter content. In addition to the well-known "axon reflex" arrangement, the capsaicin-sensitive sensory neurons have the ability to release the stored transmitter also from the same terminal which is excited by the environmental stimulus. The efferent function of these sensory neurons is realized through the direct and indirect (i.e. mediated by activation of other cells) effects of released mediators. The action of released transmitters on postjunctional elements covers a wide range of effects which may have a physiological or pathological relevance. Development of drugs capable of controlling the sensory-efferent functions of the capsaicin-sensitive sensory neurons represent a new and very promising area of research for pharmacological treatment of various human diseases.


A clinicopathological observation of Nyssen-van Bogaert syndrome with second motor neuron degeneration: two distinct clinical entities.
To report a novel clinicopathological observation of Nyssen-van Bogaert syndrome. To compare this observation with those previously reported. To discuss the nosological entity of this syndrome. To define the exact etiopathogenic mechanism of the neurogenic amyotrophy occurring at the late stage of the disease. The patient was a 16-year-old girl who developed loss of vision and deafness at the age of 8. Ataxia with slight cerebellar signs were present by the age of 14. Over the next 2 years, she developed distal weakness and wasting of the legs with depressed ankle reflexes. She died at the age of 16. Deparaffinized sections of the brain, the brain stem, the cerebellum and the spinal cord were stained with haematoxylin & eosin (H&E), Nissl, Woelcke, Bodian, periodic acid-schiff (PAS), Sudan Black and Kluver Barera. Antibodies anti-GFAP, anti-MPB and anti-neurofilaments were used for immunohistochemical stainings following the avidin-biotin-peroxydase complex (ABC) methods. The clinical pictures in our patient are similar to those previously reported in juvenile patients with optico-cochleo-dentate syndrome. Pathological study of the nervous system confirmed the diagnosis of Nyssen-van Bogaert syndrome and also showed a severe anterior horn, posterior horn and Clarke's column nerve cell degeneration with anterior root atrophy. From these clinical and pathological data, the authors suggest to include Nyssen-van Bogaert syndrome among the group of multiple system atrophy, propose to divide this syndrome into 2 forms (an early infantile form and a juvenile form) and consider that the neurogenic amyotrophy occurring at the late stage of the disease in juvenile and adult patients is mainly caused by the second motor neuron involvement.


The tryptophan catabolite or kynurenine pathway in major depressive and bipolar disorder: A systematic review and meta-analysis.
There is now evidence that affective disorders including major depressive disorder (MDD) and bipolar disorder (BD) are mediated by immune-inflammatory and nitro-oxidative pathways. Activation of these pathways may be associated with activation of the tryptophan catabolite (TRYCAT) pathway by inducing indoleamine 2,3-dioxygenase (IDO, the rate-limiting enzyme) leading to depletion of tryptophan (TRP) and increases in tryptophan catabolites (TRYCATs). To systematically review and meta-analyze central and peripheral (free and total) TRP levels, its competing amino-acids (CAAs) and TRYCATs in MDD and BD. This review searched PubMed, Google Scholar and SciFinder and included 121 full-text articles and 15470 individuals, including 8024 MDD/BD patients and 7446 healthy controls. TRP levels (either free and total) and the TRP/CAAs ratio were significantly decreased (p < 0.0001) in MDD/BD as compared with controls with a moderate effect size (standardized mean difference for TRP: SMD = -0.513, 95% confidence interval, CI: -0.611; -0.414; and TRP/CAAs: SMD = -0.558, CI: -0.758; -0.358). Kynurenine (KYN) levels were significantly decreased in patients as compared with controls with a small effect size (p < 0.0001, SMD = -0.213, 95%CI: -0.295; -0.131). These differences were significant in plasma (p < 0.0001, SMD = -0.304, 95%CI: -0.415, -0.194) but not in serum (p = 0.054) or the central nervous system (CNS, p = 0.771). The KYN/TRP ratio, frequently used as an index of IDO activity, and neurotoxicity indices based on downstream TRYCATs were unaltered or even lowered in MDD/BD. Our findings suggest that MDD and BD are accompanied by TRP depletion without IDO and TRYCAT pathway activation. Lowered TRP availability is probably the consequence of lowered serum albumin during the inflammatory response in affective disorders.


Structure-Activity Relationship Studies on 6-Chloro-1-phenylbenzazepines Leads to the Identification of a New Dopamine D1 Receptor Antagonist.
The 1-phenylbenzazepine template has yielded a number of D1R-like ligands, which, though useful as pharmacological tools, have significant drawbacks in terms of selectivity versus D5R as well as pharmacokinetic behavior. A number of 1-phenylbenzazepines contain a 6-chloro functional group, but extensive SAR studies around the 6-chloro-1-phenylbenzazepine framework have not been reported in the literature. To further understand the tolerance of the 6-chloro-1-phenylbenzazepine template for various substituent groups towards affinity and selectivity at D1R, we synthesized two series of analogs with structural variations at the C-7, C-8, N-3, C-3' and C-4' positions. The series 2 analogs differed from series 1 analogs in possessing a nitrogenated functionality at C-8 and lacked a C-4' substituent, but were otherwise similar. Analogs were assessed for affinity at D1R, D2R and D5R. For both series, we found that the analogs lacked affinity for D2R and showed modest D1R versus D5R selectivity. For series 1 analogs, an N-3 methyl substituent group was better tolerated than N-H or an N-3 allyl substituent. The C-8 position appears to be tolerant of amino and methanesulfonamide substituents for high D1R affinity, but C-8 amides displayed low to moderate D1R affinities. A C-3' methyl substituent appeared to be critical for the D1R affinity of some analogs, but the C-4' substituents tried (hydroxy and methoxy; series 1) did not result in any significant boost in D1R affinity. Compound 15a was the most potent and selective D1R ligand identified from these studies (Ki at D1R = 30 nM; 6-fold selectivity versus D5R). Further functional activity assessments indicate that 15a functions as a D1R antagonist towards cAMP-mediated signaling. The predicted drug-like properties of 15a are encouraging for further pharmacological assessments on the compound.


Childhood psychosis and monoamine metabolites in spinal fluid.
Determination of monoamine metabolites was accomplished in the spinal fluid of 22 psychotic children and in 22 sex- and almost-age-matched "normal" controls. Also, specimens from groups of mentally retarded children and children with progressive encephalopathy or meningitis were used for comparison. The psychotic children showed raised levels of homovanillic acid. Thirteen children diagnosed as autistic by Rutter's criteria showed isolated increase of this metabolite. In the group of 9 children with other psychoses, both the level of homovanillic acid and that of 5-hydroxy-indoleacetic acid was raised. The comparison with the group of "simply" mentally retarded children and results within the psychotic group revealed that the increased concentration of monoamines was not attributable to mental retardation per se.


Analysis of the Circular Transcriptome in the Synaptosomes of Aged Mice.
Recently, circular RNAs (circRNAs) have been revealed to be an important non-coding element of the transcriptome. The brain contains the most abundant and widespread expression of circRNA. There are also indications that the circular transcriptome undergoes dynamic changes as a result of brain ageing. Diminished cognitive function with increased age reflects the dysregulation of synaptic function and ineffective neurotransmission through alterations of the synaptic proteome. Here, we present changes in the circular transcriptome in ageing synapses using a mouse model. Specifically, we observed an accumulation of uniquely expressed circular transcripts in the synaptosomes of aged mice compared to young mice. Individual circRNA expression patterns were characterized by an increased abundance in the synaptosomes of young or aged mice, whereas the opposite expression was observed for the parental gene linear transcripts. These changes in expression were validated by RT-qPCR. We provide the first comprehensive survey of the circular transcriptome in mammalian synapses, thereby paving the way for future studies. Additionally, we present 16 genes that express solely circRNAs, without linear RNAs co-expression, exclusively in young and aged synaptosomes, suggesting a synaptic gene network that functions along canonical splicing activity.


The effect of Tauroursodeoxycholic acid (TUDCA) and gut microbiota on murine gallbladder stone formation.
The incidence of gallstone-related disease steadily increased in the last few years. Here, we aimed to investigate the effect of tauroursodeoxycholic acid1 (TUDCA) on preventing cholesterol gallstones formation in high-fat fed (HFD) mice. Specific pathogen-free male C57Bl/6 mice were fed a lithogenic diet2 (LD group) alone or in combination with TUDCA (5g/kg diet) for 8 weeks. Upon sacrifice, serum, gallbladder, liver and small intestine were collected and the formation of gallstones or crystals in the gallbladder was analyzed. Additionally, the intestinal microbiota, and bile acid composition, serum lipids and hepatic lipids were studied. Cholesterol gallstones with cholesterol crystals formed in mice of the LD-fed group (15/15, 100%). However, only cholesterol crystals were found in three mice without the presence of any gallstone in the TUDCA-treated group. Both serum and hepatic total cholesterol levels in the TUDCA group were significantly decreased compared with the LD group. Concomitantly, mRNA expression of Abcg5 and Abcg8 was significantly lower in the liver of the TUDCA group whilst mRNA transcripts for Abcb11, Acat2, and Cyp27 were significantly increased compared with the LD group. Additionally, the gallbladder cholesterol saturation index (1.06±0.15) in the TUDCA group was significantly decreased compared with the LD group. Interestingly, the ratio of Firmicutes/Bacteroides in the TUDCA group was increased 3x fold. TUDCA can inhibit the absorption and synthesis of lipids in the small intestine by improving the intestinal microbiota in HFD-fed mice, thus reducing gallstone formation.


The c-Ring of the F1FO-ATP Synthase: Facts and Perspectives.
The F1FO-ATP synthase is the only enzyme in nature endowed with bi-functional catalytic mechanism of synthesis and hydrolysis of ATP. The enzyme functions, not only confined to energy transduction, are tied to three intrinsic features of the annular arrangement of c subunits which constitutes the so-called c-ring, the core of the membrane-embedded FO domain: (i) the c-ring constitution is linked to the number of ions (H(+) or Na(+)) channeled across the membrane during the dissipation of the transmembrane electrochemical gradient, which in turn determines the species-specific bioenergetic cost of ATP, the "molecular currency unit" of energy transfer in all living beings; (ii) the c-ring is increasingly involved in the mitochondrial permeability transition, an event linked to cell death and to most mitochondrial dysfunctions; (iii) the c subunit species-specific amino acid sequence and susceptibility to post-translational modifications can address antibacterial drug design according to the model of enzyme inhibitors which target the c subunits. Therefore, the simple c-ring structure not only allows the F1FO-ATP synthase to perform the two opposite tasks of molecular machine of cell life and death, but it also amplifies the enzyme's potential role as a drug target.


Morphologic and molecular analysis of estrogen-induced pituitary tumorigenesis in targeted disruption of transforming growth factor-beta receptor type II and/or p27 mice.
The critical genes and products involved in estrogen-induced tumorigenesis of the pituitary gland were investigated in heterozygous transforming growth factor-beta (TGF-beta) receptor type II and p27 knockout mouse models. Tgfbr2(+/-), p27(+/-); Tgfbr2(+/-), and p27(+/-) mice and C57BL/6J wild-type mice received sc implantation of estrogen or placebo pellets for 16 or 25 wk, after which the mice were sacrificed and their pituitary glands removed for examination. The bromodeoxyuridine labeling indexes in tissue from both the anterior and intermediate pituitary lobes from p27 (+/-) and Tgfbr2(+/-); p27(+/-) mice were significantly higher than those from wild-type and Tgfbr2(+/-) mice after treatment with estrogen for 16 wk. Pituitary tumorigenesis was significantly accelerated in Tgfbr2(+/-), p27(+/-), and Tgfbr2(+/-); p27(+/-) mice compared with wild-type mice after treatment with estrogen for 16 wk. Pituitary tumorigenesis was not accelerated in Tgfbr2(+/-); p27(+/-) mice compared with Tgfbr2(+/-) or p27(+/-) mice. Expression of TGF-beta receptor type II mRNA was lower in the pituitary gland of Tgfbr2(+/-) mice than in wild-type mice before estrogen treatment and was significantly reduced after treatment. Pituitary tumorigenesis is accelerated in mice with severe TGF-beta resistance, and greatly accelerated in mice with TGF-beta resistance combined with decreased p27 expression compared with wild-type mice. Both the TGF-beta receptor type II gene and p27 gene and their products are involved in estrogen-induced tumorigenesis.


[Comparative study in nuclear and cytoplasmic RNA species of mouse liver, Guelstein ascites hepatoma 22A and liver of tumor-bearing animals].
A comparative study on nuclear and cytoplasmic DNA-like RNA species from normal liver, malignant Guelstein hepatoma 22A and liver of tumor-bearing animals was carried out. The nuclear RNA's were found to be poorly and rapidly labelling, the latter comprising a lesser proportion in hepatoma as compared with a normal liver. The comparison of labelling rates of nuclear and cytoplasmic RNA species shows that the nucleus-to-cytoplasme RNA transport is accelerated in hepatoma. Besides, cytoplasmic DNA-like RNA's are more heterogeneous in hepatoma and liver of tumor-bearing animals than in normal liver cells.


Positional dyslexia.
Position-specific errors in word reading are usually associated with neglect or visual extinction on the same side as the reading problem. In this study, two patients with left-hemisphere lesions showed visual extinction on the right but reading difficulty on the left side of words and pseudowords. Further study of one patient revealed that he also had problems reading the beginning of words presented tachistoscopically or in vertical orientation. In addition, the positional difficulty was apparent when he named the letters in words. The pattern of results indicates that the positional dyslexia in these patients was not likely attributable to general deficits in visual perception or attention but may have reflected a disorder at a later stage of letter processing.


Performance of the HPV E6/E7 mRNA Aptima HPV assay combined with partial genotyping compared with the HPV DNA Cobas 4800 HPV test for use in primary screening: Results from the CERVIVA HPV primary screening study in Ireland.
There are currently several validated HPV tests. However, longitudinal data which spans appropriate age ranges, as well as evaluation of potential screening algorithms are necessary for screening programmes choice of test. The objective of our study was to evaluate the performance of HPV mRNA and HPV DNA testing, including partial genotyping, in routine cervical screening. As part of the CERVIVA HPV Primary Screening Study, ThinPrep samples from 10 150 women were tested for HPV mRNA using the Aptima HPV assay and HPV DNA using the Cobas 4800 HPV test. HPV mRNA-positive women were further assessed with the Aptima genotyping assay for HPV 16/18/45. Baseline cytology and prospective follow-up data were collected. The performance of the two tests was examined over 42 months (to date). HPV mRNA demonstrated equivalent sensitivity to HPV DNA testing for detection of CIN2+ (93.2% [92.4-93.9] vs 92.8% [92.0-93.6], respectively) and CIN3+ (94.6% [93.8-95.3] vs 94.6% [93.8-95.3]). HPV mRNA testing had significantly higher specificity compared to HPV DNA for detection of CIN2+ (84.0% [83.5-84.5] vs 80.8% [80.2-81.4], respectively) and CIN3+ (88.44% [88.2-88.6] vs 85.62 [85.4-85.9]). The proportion of CIN2+ and CIN3+, over 3 years (42 months), in HPV-negative women was comparable for both RNA (0.20% and 0.10%) and DNA (0.22% and 0.11%). Genotyping data was comparable across both assay platforms. In the context of HPV primary screening HPV mRNA testing has potential to reduce triage tests and follow-up tests at 12 months compared to DNA testing, with no significant difference in detection of CIN2+ and CIN3+.


Inhibition of photosensitized degradation of organic contaminants by copper under conditions typical of estuarine and coastal waters.
Dissolved organic matter (DOM) driven-photochemical processes play an important role in the redox cycling of trace metals and attenuation of organic contaminants in estuarine and coastal ecosystems. In this study, we evaluate the effect of Cu on 4-carboxybenzophenone (CBBP) and Suwannee River natural organic matter (SRNOM)-photosensitized degradation of seven target contaminants (TCs) including phenols and amines under pH conditions and salt concentrations typical of those encountered in estuarine and coastal waters. Our results show that trace amounts of Cu(II) (25 -500 nM) induce strong inhibition of the photosensitized degradation of all TCs in solutions containing CBBP. The influence of TCs on the photo-formation of Cu(I) and the decrease in the lifetime of transformation intermediates of contaminants (TC•+/ TC•(-H)) in the presence of Cu(I) indicated that the inhibition effect of Cu was mainly due to the reduction of TC•+/ TC•(-H) by the photo-produced Cu(I). The inhibitory effect of Cu on the photodegradation of TCs decreased with the increase in Cl- concentration since less reactive Cu(I)-Cl complexes dominate at high Cl- concentrations. The impact of Cu on the SRNOM-sensitized degradation of TCs is less pronounced compared to that observed in CBBP solution since the redox active moieties present in SRNOM competes with Cu(I) to reduce TC•+/ TC•(-H). A detailed mathematical model is developed to describe the photodegradation of contaminants and Cu redox transformations in irradiated SRNOM and CBBP solutions.


AT(2) but not AT(1) receptor antagonism abolishes angiotensin II increase of the acquisition of conditioned avoidance responses in rats.
In this study we attempted to determine behavioural, including cognitive, consequences of the brain AT(1) (losartan, 2 nmol), AT(2) (PD 123319, 1.5 nmol), and joint AT(1)/AT(2) angiotensin receptors blockade. Male Wistar rats (160-180 g) were injected into the left cerebral ventricle with the above doses of the blockers dissolved in 0.9% NaCl solution (vehicle) or with the vehicle alone. Five minutes later they received, to the right cerebral ventricle, 1 nmol of angiotensin II (Ang II) dissolved in vehicle or the vehicle alone. Ang II consistently increased rate of acquisition of conditioned avoidance response (CARs) and facilitated recall of the passive avoidance behaviour. In one out of the three series of experiments in open field Ang II stimulated rats locomotor activity. Losartan and PD 123319, both ineffective alone, given prior to Ang II abolished all the behavioural changes produced by the peptide except for the Ang II facilitation of CARs acquisition, which was unchanged by losartan. Interestingly, joint injection of losartan and PD 123319 significantly decreased the rate of CARs acquisition both in control and Ang II treated animals. In conclusion, the present data suggest significant though different involvement of both AT(1) and AT(2) angiotensin receptors in cognitive processes.


The Effect of Platelet-Rich Fibrin Matrix on Skin Rejuvenation: A Split-Face Comparison.
Despite the increasingly popular use of platelet-rich fibrin matrix (PRFM) for facial rejuvenation, no studies have evaluated its effects on skin. The aim of this study was to determine the effect of PRFM on skin quality as measured by the Canfield VISIA Complexion Analysis System. This was a 12-week randomized, placebo-controlled trial conducted at a single center comparing 2 groups. Thirty patients received a PRFM injection in the mid-cheek region and nasolabial fold on one side of the face and saline on the contralateral side. The primary outcome measure was the difference between pre- and posttreatment total VISIA skin scores for each group at 6 and 12 weeks. The change in subscores for each skin parameter was also calculated. There were 30 participants, with a mean [standard deviation] age of 49.9 [13.9] years. At 6 weeks, the median change in total VISIA score (interquartile range) was -1.77 (2.36) in the PRFM group and -0.73 (2.09) in the saline group (P = 0.003). The only skin parameter that had a significantly different change in score between the groups was texture (P = 0.004). At 12 weeks, the change in median score was -1.31 (3.26) in the PRFM cohort and -0.76 (2.21) in the saline cohort (P = 0.34). There was no statistical significance in the change in score for any of the individual skin parameters. PRFM can objectively improve skin quality compared with placebo. Texture was the only skin parameter that significantly improved, which is consistent with PRFM's role as a filler agent. The results appear to persist for at least 6 weeks.


Paraplegia after thoracotomy under combined general and epidural anesthesia in a child.
A 9-year-old boy underwent a thoracotomy for excision of his right third rib under combined general and epidural anesthesia for a Ewings sarcoma. Postoperatively, he was found to have a complete T2-3 paraplegia. Permanent paraplegia was described as a rare complication of thoracotomy in adults, and very rarely after epidural analgesia in adults and babies. This was the first report in a child.


Stability of selected microRNAs in human blood, semen and saliva samples exposed to different environmental conditions.
Body fluid identification is an essential aspect of forensic investigations. MicroRNAs (miRNAs), a type of non-coding RNA has a lot of potential in forensic studies as biomarkers. Using a reference gene - GAPDH, this study ascertained the stability of hsa-mir-451a, hsa-miR-10b, and hsa-miR-205 in blood, semen and saliva respectively, exposed to different environmental conditions over different periods of time. Twenty adult males aged 20-35 donated blood, sperm, and saliva. These fluids were separated into four groups and exposed to Outdoor, Indoor, Fridge, and Freezer conditions for 37 days. Total RNA was isolated using Trizol-Isopropanol extraction protocol. FIREScript RT cDNA synthesis kit and Luna Universal qPCR Master Mix were used for cDNA synthesis and qPCR respectively. Under different environmental conditions, miR-451a and miR-10b was detected in all blood and semen samples but not in all saliva samples. Pearson's correlation test for Cq values showed multiple strong correlations (p < 0.05) within and outside the groups compared to GADPH. Regardless of the body fluid sample, the freezer group was more stable compared to other groups, although correlations are not homogeneous in all samples. Expression of the targeted genes in the body fluids showed that hsa-miR-451a, hsa-miR-10b, hsa-miR-205 could be utilized as biomarkers in forensics for body fluids identification.


Resolution and properties of the catalytic unit of cardiac adenylate cyclase.
The catalytic unit (C) of cardiac adenylate cyclase was resolved from the guanine nucleotide-binding regulatory protein (G/F) by fractionation of Lubrol 12A9 extracts with 33% saturated (NH4)2SO4 and by gel filtration in the presence of 3-[(3-cholamidopropyl)dimethylammonio]-l-propanesulfonate (CHAPS). Catalytic activity in both preparations was supported by Mg2+ and Mn2+, Vmax values being 12 fold higher in the presence of the latter cation. NaF-activated preparations were also subjected to gel filtration in the presence of CHAPS. In this case, both the catalytic activity and G/F emerged in the activated state. G/F could be deactivated by dialysis of the preparation in the absence of NaF; the catalyst however remained activated following dialysis. NaF sensitivity was reconstituted in C by nonactivated and NaF-activated preparations of G/F isolated by gel filtration. Reconstitution was dependent upon the amount of both G/F and C in the assay. Nonactivated G/F also reconstituted guanine nucleotide sensitivity in C. Catalytic activity was thermally labile, but was stabilized at 25 degrees C by substrate (Mn2+ ATP). C was stimulated up to 25-fold by forskolin. The NaF-activated catalyst resolved by gel filtration was relatively insensitive to this agent. Forskolin, however, augmented NaF-sensitivity by both non-activated and NaF-activated G/F provided it was added to the assay before G/F. Similarly, forskolin augmented guanine nucleotide sensitivity of nonactivated G/F in the presence of GTP gamma S. The P site agent 2',5'-dideoxyadenosine (DDA) was a weak inhibitor of nonactivated C, but a powerful inhibitor of C stimulated by forskolin or activated by G/F in the presence of NaF. In all respects C precipitated by (NH4)2SO4 appeared to be identical with that resolved by gel filtration. Ammonium sulfate precipitation, because of its simplicity, speed, relatively good yield, and adaptability for large scale operation, may be the preferred method for preparing cardiac C for purification studies


Conflicts of interest in infection prevention and control research: no smoke without fire. A narrative review.
Conflicts of interest (COIs) do occur in healthcare research, yet their impact on research in the field of infection prevention and control (IPC) is unknown. We conducted a narrative review aiming to identify examples of COIs in IPC research. In addition to well-known instances, we conducted PubMed and Google searches to identify and report case studies of COIs in IPC and antimicrobial resistance (AMR), which were chosen arbitrarily following consensus meetings, to illustrate different types of COIs. We also searched the Retraction Watch database and blog to systematically identify retracted IPC and/or infectious disease-related papers. Our review highlights COIs in academic research linked to ties between industry and physicians, journal editors, peer-reviewed journals' choice for publication, and guideline committees participants and authors. It explores how COIs can affect research and could be managed. We also present several selected case studies that involve (1) the chlorhexidine industry and how it has used marketing trials and key opinion leaders to promote off-label use of its products; (2) the copper industry and how reporting of its trials in IPC have furthered their agenda; (3) the influence of a company developing "closed infusion systems" for catheters and how this affects networks in low- and middle-income countries and guideline development; (4) potential perverse incentives hospitals may have in reporting healthcare-associated infection or AMR rates and how government intervention may restrict AMR research for fear of bad publicity and subsequent negative economic consequences. Finally, the analysis of reasons for the retraction of previously published papers highlights the fact that misconduct in research may have other motivations than financial gain, the most visible form of COIs. COIs occur in the field of research in general, and IPC and AMR are no exceptions. Their effects pervade all aspects of the research and publication processes. We believe that, in addition to improvements in management strategies of COIs, increased public funding should be available to decrease researchers' dependency on industry ties. Further research is needed on COIs and their management.


The effects of over-expression and suppression of Cdc25A on the S-phase checkpoint induced by benzo(a)pyrene.
Our previous results have indicated that Cdc25A is involved in benzo(a)pyrene (BaP)-induced S-phase checkpoint in 16HBE cells and A549 cells. In this paper, we reported the changes of the downstream molecular pathway of Cdc25A and the effects of over-expression and suppression of Cdc25A on BaP-induced S-phase checkpoint. In the S-phase checkpoint induced by BaP the reduction of Cdc25A contributes to cyclin A inhibition. Over-expression of Cdc25A abrogated BaP-induced S-phase arrest in 16HBE cells and concomitantly the expression levels of Cdk2 and cyclin A were not obviously changed by BaP when compared with the control. Cdc25A down-regulation by RNA interference (RNAi) prolonged the S-phase arrest induced by BaP and decreased clearly the expression levels of cyclin A and cyclin E. Therefore, our results further demonstrated that Cdc25A was an effector in Chk1-Cdc25A-cyclin A/Cdk2 pathway of S-phase checkpoint elicited by the carcinogen BaP in 16HBE cells.


A BALB/c mouse model for assessing the potential allergenicity of proteins: comparison of allergen dose, sensitization frequency, timepoint and sex.
The present study was to investigate the possibility of using the BALB/c mouse as an animal model for assessing the potential allergenicity of proteins. Specific IgE and IgG1 against ovalbumin were induced by dosing BALB/c mice via intraperitoneal injection (absence of adjuvant). The effects of various allergen doses (5 mg, 0.5 mg or 0.05 mg OVA), sensitization times (twice or five times), timepoints (day 14 or day 28) and sex (male or female) were studied. IL-4, IFN-γ, OVA-specific IgE and IgG1 were measured by enzyme-linked immunosorbent assay (ELISA). A general finding was that mice treated with 0.05 mg OVA had the highest OVA-specific IgE and IgG1, statistically significant higher specific IgE and IgG1 were observed in groups sensitized five times than twice, OVA-specific IgE and IgG1 on day 28 were statistically higher than day 14, and higher IL-4 was observed in OVA-allergic mice than control mice. These results demonstrate that the BALB/c mouse model treated with 0.05 mg OVA intraperitoneally on days 0, 3, 6, 9, 12 might be used for further experiments. OVA-specific IgE and IgG1 should be detected on day 28. Further studies including reproducibility and other conditions were required before using the BALB/c mouse model for assessing the potential allergenicity of proteins.


Charcot-Marie-Tooth neuropathy due to a novel EGR2 gene mutation with mild phenotype--usefulness of human mapping chip linkage analysis in a Czech family.
Charcot-Marie-Tooth neuropathies (CMT) are a group of clinically and genetically heterogeneous disorders of the peripheral nervous system. Selection of candidate disease genes for mutation analysis is sometimes difficult since more than 40 genes and loci are known to be associated with CMT neuropathies. Hence a Czech family Cz-CMT with demyelinating type of autosomal dominant CMT disease was investigated by genome-wide linkage analysis by means of single-nucleotide polymorphism (SNP) arrays. Among 35 regions with linkage, five carried known CMT genes. In the final result a novel early growth response 2 - missense mutation c.1235 A>G, p.Glu412Gly was found. Surprisingly, the more severely affected proband carried an additional heterozygous myelin protein zero variant p.Asp246Asn detected previously, which may modify the phenotype. However, this MPZ variant is benign in heterozygous state alone, because it is also carried by the patient's healthy father.


A hybrid patch clamp amplifier.
The current-to-voltage convertor used in patch clamping is analyzed for noise generation and the major noise sources determined. A hybrid patch clamp amplifier design is theoretically analyzed. Here it is shown that by differentiation and recombining of signals the original input signal can be reconstructed. Several circuits of this design are described and their performance compared. The optimal signal detection obtained for a 1 ms current pulse width with a signal-to-noise ratio of 1 is 0.025 pA. In these circuits, high frequency attenuation is readily accomplished with a single control. In addition, compensation for the transients which occur with step control voltages is effectively accomplished. With this circuitry, it is shown that for most patch clamp situations, the minimum pulse width and current which can be detected is determined by the patch clamp seal resistance.


Primary tumour resection in metastatic nonfunctioning pancreatic endocrine carcinomas.
The role of debulking surgery in metastatic nonfunctioning pancreatic endocrine carcinomas (M-NF-PECs) with resectable primary tumour and unresectable liver metastases is debated. Aim of the study is to evaluate whether the resection of the primary tumour in metastatic nonfunctioning pancreatic endocrine carcinoma improves survival. Fifty-one metastatic nonfunctioning pancreatic endocrine carcinoma patients with unresectable liver metastases were enrolled from 1990 to 2004 at the time of diagnosis. Nineteen patients underwent complete resection of the primary tumour whilst 32 were judged unresectable. All cases were classified according to the WHO 2000 classification. All clinico-pathological parameters, including grade of differentiation and the Ki-67 proliferation index were considered in univariate and multivariate models. Of the 19 resected patients, 14 (73.7%) underwent left-pancreatectomy and 5 (26.3%) pancreaticoduodenectomy. In the unresected group of 32 patients, 9 (28.1%) underwent surgical biliary and/or gastric by-pass. There was no postoperative mortality and the median survival was 54.3 months (95% CI: 25.7-82.9). No difference in survival was observed between the two groups [resected: median 54.3 months (95% CI: 25-83.6), unresected: median 39.5 months (95% CI: 5.4-73.6); p=0.74]. Upon multivariate analysis poor differentiation (HR 3.01; 95% CI 1.08-8.4; p=0.035) and a Ki-67 index > or = 10% (HR 4.4; 95% CI 1.2-16.1; p=0.023) were significant predictors of survival. Resection of the primary pancreatic tumour in metastatic nonfunctioning pancreatic endocrine carcinoma patients with unresectable liver metastases does not significantly improve survival. Resection can be considered as symptomatic palliative therapy in patients with well-differentiated endocrine carcinomas and a proliferative index lower than 10%.


Chromosomal sex-determining regions in animals, plants and fungi.
The independent evolution of sex chromosomes in many eukaryotic species raises questions about the evolutionary forces that drive their formation. Recent advances in our understanding of these genomic structures in mammals in parallel with alternate models such as the monotremes, fish, dioecious plants, and fungi support the idea of a remarkable convergence in structure to form large, non-recombining regions with discrete evolutionary strata. The discovery that evolutionary events similar to those that have transpired in humans have also occurred during the formation of sex chromosomes in organisms as divergent as the plant Silene, the fungus Cryptococcus and the fish medaka highlights the importance of future studies in these systems. Such investigation will broaden our knowledge of the evolution and plasticity of these ubiquitous genomic features underlying sexual dimorphism and reproduction.


Th17 cells and their cytokines serve as potential therapeutic target in experimental autoimmune neuritis.
Accumulating evidence has pointed that T helper 17 cells and their cytokines are pathogenic in Guillain-Barré syndrome (GBS). However, little is known concerning the IL-17 expression change trend during the whole course of disease, and whether drugs specially targeting Th17 cells or their cytokines have potential effects on experimental autoimmune neuritis (EAN) is uncertain. We explored the IL-17 and receptor-related orphan receptor-gamma-t (RORγt) expression change trends in EAN rats to identify the stage of effect of Th17 pathway in EAN, and further, we investigated the effect of RORγt inhibitors by assessing clinical score, histological staining, and IL-17 and RORγt expression change trends in serum and tissues. The expression level of IL-17 and RORγt in serum and tissues increased with the progression of the disease in the EAN group and decreased after the disease reaching its peak. RORγt-IN-1 treatment strikingly reduced the neurological deficits by ameliorating inflammatory cell infiltration, deceased the serum IL-17 and RORγt levels, and further downregulated the expression of IL-17 and RORγt mRNA in spleen, lymphnodes, and sciatic nerve. Th17 cells and their cytokines are closely associated with the onset of GBS and the novel RORγt inhibitors may be prospective strategies in treating GBS.


The chicken with four legs: a case of semantic amnesia and cryptogenic epilepsy.
Here we describe the unusual condition of selective semantic amnesia related to cryptogenic temporal lobe epilepsy. XY, an adult male patient, presented with partial seizures and disabling dysnomia. Neuropsychological tests revealed seriously impaired semantic memory. Electroencephalography documented ictal epileptic abnormalities in the left temporal lobe. Positron emission tomography showed reduced metabolism in the temporoparietal regions, but the results of magnetic resonance imaging and spectroscopy were normal. After 11 years, neuropsychological tests showed selective impairment of semantic memory and computed tomography provided normal results. This case shows that semantic memory may be selectively and lastingly altered, highlighting a distinction between semantic amnesia and global amnesia. Moreover, it is unique in that it occurred without evidence of gross temporal lobe pathology. The pathophysiological pattern of epileptic abnormalities in the left temporoparietal cortex supports the role played by dysfunctional neuronal networks (as provoked by focal epileptic discharges) in determining selective semantic amnesia.


Physical Activity Level of Korean Adults with Chronic Diseases: The Korean National Health and Nutritional Examination Survey, 2010-2012.
Proper physical activities are known to be helpful in the prevention and management of chronic diseases. However, the physical activity level of patients with chronic diseases is low. Therefore, this study aimed to investigate the physical activity compliance of patients with hypertension, diabetes, and dyslipidemia in Korea. This study analyzed the 2010-2012 Fifth Korean National Health and Nutrition Examination Survey data. We included 13,873 individuals in the analysis. The level of physical activity compliance was measured by performing multivariate logistic regression analyses. In the univariate analysis, the subjects with hypertension or diabetes tended to comply with the physical activity guidelines less faithfully than their healthy counterparts. The proportion of subjects with hypertension who were insufficiently physically active was 65.4% among the men and 75.8% among the women. For diabetes, the proportions were 66.7% and 76.8%, respectively. No significant difference was found between the subjects with dyslipidemia and their healthy counterparts. In the multivariate logistic regression analysis, no significant difference in physical activity compliance was observed between the subjects with hypertension, diabetes, or dyslipidemia and their healthy counterparts for both sexes. The patients with hypertension or diabetes tended to have lower physical activity prevlaence than their healthy counterparts. However, for dyslipidemia, no significant difference was found between the two groups. Given the significance of physical activities in the management of chronic diseases, the physical activities of these patients need to be improved.


Large epidermal cyst of the clitoris: a novel diagnostic approach to assist in surgical removal.
Epidermal cysts of the clitoris are uncommon and usually associated with previous trauma. Surgical removal is the preferred treatment with close attention to preserve sensation and vascular supply. The use of an alternative coil during Magnetic Resonance (MR) imaging to visualize the anatomy may improve surgical management and outcomes. A 15-year-old adolescent had a 2-year history of clitoral enlargement. Physical exam revealed a 4 × 5 cm clitoral cyst and normal hormonal evaluation. Initial MR with a pelvic coil showed poor resolution of neurovascular supply. MR was repeated using a small flex coil (commonly used on the wrist) placed as a sanitary napkin providing a higher resolution image of the neurovascular structures, which guided surgical intervention. The use of the MR small flex coil to better visualize the external genital structures and clitoral anatomy improved preoperative evaluation and guided surgical management of the clitorial cyst.


Effect of Different Tensoactives on the Morphology and Release Kinetics of PLA-b-PEG Microcapsules Loaded With the Natural Anticancer Compound Perillyl Alcohol.
Perillyl alcohol is a natural compound that has attracted a significant interest due to its potent antitumor activity. However, clinical trials have exhibited poor tolerance by oral administration, mainly due to gastrointestinal side effects. We propose the entrapment of perillyl alcohol into poly(D,L-lactic acid)-block-poly(ethylene glycol) (PLA-b-PEG) as delivery platform (entrapment efficiency of 63%-68%). The influence of different concentrations of the tensoactives poly(vinyl alcohol) and sodium cholate (SC) on shear strength and morphology was evaluated by confocal laser scanning microscopy and interfacial tension studies. Only the microcapsules formulated with SC maintained their sphericity when submitted to shear stress. These results indicate that the interface is better organized with SC, conferring mutual stacked packing that is able to better stabilize the organic drop. The in vitro release profile of the drug from the microcapsules was correlated with pore formation and polymer degradation, best fitted to the Baker-Lonsdale model. The loaded microcapsules showed an IC50 equivalent to that of the free drug (80 μg/mL) after 72 h of exposure. However, after 24 h of exposure, loaded microcapsules showed an IC50 almost two-fold higher (220 μg/mL) suggesting gradual release.


Management of constipation and encopresis in children.
Constipation is a common problem in primary care. Nurse practitioners should be able to diagnose and treat constipation appropriately and to recognize which children require referral to a gastroenterologist. Referral to a gastroenterologist is necessary for the child with simple constipation or encopresis fails to respond to treatment, an organic etiology is suspected, or complex management is required.


Astrocytes in the spinal cord contributed to acute stress-induced gastric damage via the gap junction protein CX43.
Rat restraint water-immersion stress (RWIS) is a compound stress of high intensity and is widely used to study the pathological mechanisms of stress gastric ulcers. The spinal cord, as a part of the central nervous system, plays a dominant role in the gastrointestinal tract, but whether the spinal cord is involved in rat restraint water-immersion stress (RWIS)-induced gastric mucosal damage has not been reported. In this study, we examined the expression of spinal astrocytic glial fibrillary acidic protein (GFAP), neuronal c-Fos, connexin 43 (Cx43), and p-ERK1/2 during RWIS by immunohistochemistry and Western blotting. In addition, we intrathecally injected the astrocytic toxin L-a-aminoadipate (L-AA), gap junction blocker carbenoxolone (CBX), and ERK1/2 signaling pathway inhibitor PD98059 to explore the role of astrocytes in the spinal cord in RWIS-induced gastric mucosal damage and its possible mechanism in rats. The results showed that the expression of GFAP, c-Fos, Cx43, and p-ERK1/2 was significantly elevated in the spinal cord after RWIS. Intrathecal injection of both the astrocyte toxin L-AA and the gap junction blocker CBX significantly attenuated RWIS-induced gastric mucosal damage and decreased the activation of astrocytes and neurons induced in the spinal cord. Meanwhile, the ERK1/2 signaling pathway inhibitor PD98059 significantly inhibited gastric mucosal damage, gastric motility and RWIS-induced activation of spinal cord neurons and astrocytes. These results suggest that spinal astrocytes may regulate the RWIS-induced activation of neurons via CX43 gap junctions and play a critical role in RWIS-induced gastric mucosa damage through the ERK1/2 signaling pathway.


Reduction in FSH receptors in the rat testis by injection of homologous hormone.
Intratesticular injection of 25 microgram rat FSH into rats under continuous urethane anaesthesia resulted 24 h later in a 50% reduction in binding sites for FSH in testicular homogenates. By 48 h after injection, receptor number usually returned to control values. Intratesticular injection of 125I-labelled rat RSH showed less than 1% remaining in the testis 24 h later, suggesting that the reduction in receptor numbers at 24 h is not due to occupancy by the FSH. Experiments did not suggest that the injection of FSH induced FSH-degrading enzymes or inhibitors of binding.