Characterization of new G protein-coupled adenine receptors in mouse and hamster.
The nucleobase adenine has previously been reported to activate G protein-coupled receptors in rat and mouse. Adenine receptors (AdeR) thus constitute a new family of purine receptors, for which the designation "P0-receptors" has been suggested. We now describe the cloning and characterization of two new members of the AdeR family from mouse (MrgA10, termed mAde1R) and hamster (cAdeR). Both receptors were expressed in Sf9 insect cells, and radioligand binding studies were performed using [(3)H]adenine. Specific binding of the radioligand was detected in transfected, but not in untransfected cells, and K D values of 286 nM (mAde1R, B max 1.18 pmol/mg protein) and 301 nM (cAdeR, B max 17.7 pmol/mg protein), respectively, were determined. A series of adenine derivatives was investigated in competition binding assays. Minor structural modifications generally led to a reduction or loss of affinity, with one exception: 2-fluoroadenine was at least as potent as adenine itself at the cAdeR. Structure-activity relationships at all AdeR orthologs and subtypes investigated so far were similar, but not identical. For functional analyses, the cAdeR was homologously expressed in Chinese hamster ovary (CHO) cells, while the mAde1R was heterologously expressed in 1321N1 astrocytoma cells. Like the previously described AdeRs from rat (rAdeR) and mouse (mAde2R), the mAde1R (EC50 9.77 nM) and the cAdeR (EC50 51.6 nM) were coupled to inhibition of adenylate cyclase. In addition, the cAdeR from hamster expressed in CHO cells produced an increase in intracellular calcium concentrations (EC50 6.24 nM) and was found to be additionally coupled to Gq proteins.


Access to public health services and integral care for women during the puerperal gravid period period in Ceará, Brazil.
Over time, the Brazilian health system, a growing country, has been developing to ensure good accessibility to health goods and services. This development is focusing on the principle of universality of access and completeness of health care. In this context, we aimed to evaluate the completeness of care and universality of access for women in their pregnancy and puerperal period in Ceará, Brazil. A descriptive, cross-sectional study based on a quantitative approach, using information collected from the database of the regulation system of the state of Ceará and data from the Prenatal Monitoring System. The research population comprised of 1701 women who delivered a baby in an obstetric reference unit in the Health Macro-Region of Cariri, Ceará, Brazil from January to December 2015. There was a high rate of cesarean delivery (49.7%) and a high waiting time for access to high-risk delivery (32.6%) and neonatal intensive care unit (72.9%). There was also a low percentage (41.1%) of pregnant women undergoing an adequate number of prenatal consultations, dental care (20%), educational activities (15%), visits to the maternity ward (0.1%), laboratory tests of the third trimester (29.2%) and puerperal consultation (37.9%). It was concluded that the Maternal and Child Health Policy, especially the Rede Cegonha, which is still under development, does not ensure access and completeness of care for women during the prenatal, delivery, and puerperal periods, thus violating their reproductive rights. The results of this study allow a critical analysis by the academia and health managers in search of strategies to improve the services of Rede Cegonha in Brazil.


Sonographic findings in leiomyoma of postorchiectomy scrotum.
Leiomyomas of the scrotum are unusual testicular masses and a case report reviewing the ultrasonographic findings of this entity comprises this report. The ultrasonographic characteristics of leiomyoma have not been reported previously. The purpose of this case report is to discuss the findings of ultrasound in evaluation of leiomyoma in the area of the scrotum.


Confocal microscopy as a tool to reveal the tridimensional organization of intracellular lumens and intercellular cysts in a human colon adenocarcinoma cell line.
Adenocarcinoma cells often form intracellular lumens and intercellular cysts. In order to study the structural relationships between these lumens and the apical domain of normal enterocytes, we have applied electron microscopy and confocal microscopy to a cloned cell line derived from the human colon adenocarcinoma cell line LoVo which express a high number of intracellular lumens and intercellular cysts. Microvilli reminiscent of those detected in the brush border of small intestinal cells are formed in the two types of compartments. By immunofluorescence, we found that a 135 kDa membrane glycoprotein characterized by a monoclonal Ab and normally associated with the brush-border of enterocytes is expressed at the surface of the intracellular lumens and intercellular cysts present in the adenocarcinoma cells. Comparison of fluorescence and reflection contrast micrographs obtained by confocal microscopy demonstrate the presence of spherical intracellular lumens in the juxtanuclear region of single cells, and of more complex shaped intercellular cysts located within clusters of cells. The later cells form junctional complexes limiting an apical plasma membrane domain in contact with the intercellular cyst. It is suggested that the intracellular lumens may represent the abortive form of an apical plasma membrane due to the lack of components required to establish epithelial cell contacts. As opposed to conventional fluorescence microscopy, confocal microscopy allows rapid inspection of the tridimensional organization of intracellular lumens and intercellular cysts even when they are located in cell multilayers.


The accuracy of parental concern expressed in the Ages and Stages Questionnaire to predict developmental delay.
Whilst there have been comparisons of the Ages and Stages Questionnaire (ASQ) with other gold standard developmental assessments, research concerning the free-text comments section of the questionnaire has been limited. The objective of this paper is to assess whether parental concerns expressed in the free-text comments section of the ASQ can accurately predict infant development. Data from the Development After Infant Surgery study was analysed post hoc. Free-text comments from the ASQ were analysed qualitatively by thematic analysis and quantitatively in comparison to the Bayley Scales of Infant and Toddler Development, Third Edition (BSID-III). Infants who underwent cardiac surgery (97), non-cardiac surgery (131) and no surgery (controls; 166) were analysed. Parental concern expressed on the ASQ predicted a lower mean BSID-III score when groups were analysed collectively. Medical concern and developmental concern were shown to be predictors of child development, whilst global concerns and behavioural concerns were unrelated to child development. The presence of carer concern was assessed for its value in predicting a BSID-III subscale score less than 8 in all groups. Sensitivity ranged from 22% to 48%, and specificity ranged from 73% to 88%. Parental concern was found to predict a lower BSID-III when all groups were analysed collectively. Despite this, parental concern as a screening test for developmental delay failed to meet the National Health and Medical Research Council guidelines for adequate psychometric properties. The findings from this study suggest that there is value in attending to the comments written by carers on the ASQ.


Adult acute lymphoblastic leukaemia in Denmark. A national population-based retrospective study on acute lymphoblastic leukaemia in Denmark 1998-2008.
Since July 2008, children and adults 1-45 years, diagnosed with acute lymphoblastic leukaemia (ALL) in Denmark have been treated according to the common Nordic Society for Paediatric Haematology and Oncology ALL2008 protocol. To explore whether this strategy will improve survival compared with historical controls, we performed a retrospective national population-based study of adult ALL between 1998 and 2008. Patients were identified through the Danish Patobank and the Danish Cancer Registry; data was collected from patient files, and included 277 patients (median age, 47 years, range 15-91 years). The 5-year projected event-free survival (pEFS(5y)) and overall survival (pOS(5y)) for the whole cohort was 27·5% [95% confidence interval (CI) 22·4-33·6] and 34·1% (95% CI 28·7-40·4), respectively. No patient above 65 years survived beyond 5 years from diagnosis. For patients receiving curatively intended treatment, the pEFS(5y) and pOS(5y) were 36·6% and 44·1%, respectively, with a significantly higher pOS(5y) for patients 15-35 years compared with patients 36-65 years (50·7% vs. 38·9%, P = 0·006). Cox multiple regression analysis identified age (Hazard Ratio = 1·7, P < 0·006) as a statistically significant predictor of EFS. The cure rates, not least for the elderly, are unacceptably low, and call for new strategies in the treatment of adult ALL in all age groups.


The development of an instrument measuring unintentional injuries in young children in low-income settings to serve as an evaluation tool for a childhood home injury prevention program.
Responding to the lack of standardized instrumentation, especially as applied in low-income contexts, the researchers decided that an instrument should be developed to serve as an evaluation tool for a childhood (unintentional) injury prevention program as well as a tool from which injury risks (poisonings, burns and falls) could be identified within households. Cross-sectional samples of 521 households in four low-income sites in Cape Town and Johannesburg, South Africa, participated in the three phases of the study. The generation of an item pool based on a table of specifications, subsequent scientific item reduction procedures, reviews from experts and pilot tests were used to develop the scales for measuring the injury risks. The developed instrument complies with all the requirements for a valid and reliable measurement instrument. The instrument allows valid comparison of risks between communities, as well as before and after comparisons for an intervention program. The provision of this instrument may bring enormous benefits to research studies. It can also provide proactive, rather than reactive, information about injury risks before they develop into injuries and thus allows focusing of safety efforts for improvement of problematic areas in the households.


Oscillations and waves of cyclic AMP in Dictyostelium: a prototype for spatio-temporal organization and pulsatile intercellular communication.
The amoebae Dictyostelium discoideum aggregate after starvation in a wavelike manner in response to periodic pulses of cyclic AMP (cAMP) secreted by cells which behave as aggregation centers. In addition to autonomous oscillations, the cAMP signaling system that controls aggregation is also capable of excitable behavior, which consists in the transient amplification of suprathreshold pulses of extracellular cAMP. Since the first theoretical model for slime mold aggregation proposed by Keller and Segel in 1970, many theoretical studies have addressed various aspects of the mechanism and function of cAMP signaling in Dictyostelium. This paper presents a brief overview of these developments as well as some reminiscences of the author's collaboration with Lee Segel in modeling the dynamics of cAMP relay and oscillations. Considered in turn are models for cAMP signaling in Dictyostelium, the developmental path followed by the cAMP signaling system after starvation, the frequency encoding of cAMP signals, and the origin of concentric or spiral waves of cAMP.


[Retroperitoneal vascular injuries (author's transl)].
From 1973-1977, 21 retroperitoneal vascular injuries were seen at the Medical School of Hannover. Vascular lesions of polytraumatized patients were treated 12 times. The rest were iatrogenic traumas after laminectomy, herniotomy, and hip replacement as well as after urologic and gynecologic operations. The overall mortality was 43%. Surgical treatment of the different vascular regions is discussed.


Depression, anxiety, stress levels, and coping styles of caregivers of patients with left ventricular assist device.
To determine the depression, anxiety, stress (DAS) levels, and coping styles of caregivers of patients with left ventricular assist device (LVAD). The sample of the study consisted of 74 caregivers of patients with LVAD. Data were collected using Individual Data Form, DASS 21, and Coping Styles Scale. Depression was found in 67.6%, anxiety in 45.9%, and stress in 50% of the caregivers of patients with LVAD. A significant relationship was found between the sociodemographic characteristics of caregivers, some variables related to the patient and the care process, and coping styles with stress, and DAS levels of caregivers. Considering the variables that may affect the DAS levels of the LVAD caregivers will improve the expected results from the LVAD.


Diols, α-Ketols, and Diones as 22π Components in [2+2+2] Cycloadditions of 1,6-Diynes via Ruthenium(0)-Catalyzed Transfer Hydrogenation.
The first use of vicinal diols, ketols, or diones as 22π components in metal-catalyzed [2+2+2] cycloaddition is described. Using ruthenium(0) catalysts, 1,6-diynes form ruthenacyclopentadienes that engage transient diones in successive carbonyl addition. Transfer hydrogenolysis of the resulting ruthenium(II) diolate mediated by the diol or ketol reactant releases the cycloadduct with regeneration of ruthenium(0) and the requisite dione.


Comparison of three cognitive assessment methods in post-stroke aphasia patients.
The cognitive level of post-stroke aphasia (PSA) patients is generally lower than non-aphasia patients, and cognitive impairment (CI) affects the outcome of stroke. However, for different types of PSA, what kind of cognitive assessment methods to choose is not completely clear. We investigated the Montreal Cognitive Assessment (MoCA), the Mini-Mental State Examination (MMSE), and the Non-language-based Cognitive Assessment (NLCA) to observe the evaluation effect of CI in patients with fluent aphasia (FA) and non-fluent aphasia (NFA). 92 stroke patients were included in this study. Demographic and clinical data of the stroke group were documented. The language and cognition were evaluated by Western Aphasia Battery (WAB), MoCA, MMSE, and NLCA. PSA were divided into FA and NFA according to the Chinese aphasia fluency characteristic scale. Pearson's product-moment correlation coefficient test and multiple linear regression analysis were performed to explore the relationship between the sub-items of WAB and cognitive scores. The classification rate of CI was tested by Pearson's Chi-square test or Fisher's exact test. The scores of aphasia quotient (AQ), MoCA, MMSE, and NLCA in NFA were lower than FA. AQ was positively correlated with MoCA, MMSE, and NLCA scores. Stepwise multiple linear regression analysis suggested that naming explained 70.7% of variance of MoCA and 79.9% of variance of MMSE; comprehension explained 46.7% of variance of NLCA. In the same type of PSA, there was no significant difference in the classification rate. The classification rate of CI in NFA by MoCA and MMSE was higher than that in FA. There was no significant difference in the classification rate of CI between FA and NFA by NLCA. MoCA, MMSE, and NLCA can be applied in FA. NLCA is recommended for NFA.


An improved algorithm for quantifying real-time impedance biosensor signals.
In previously published work [1] we presented a real-time electrochemical impedance biosensor prototype system and a state-space estimation algorithm for signal quantification. Experiments in the interim have revealed some algorithm failure modes which reduced the reliability and repeatability of quantification. The present work describes a related algorithm that introduces constraints based on a priori knowledge of the expected signals predicted by the biosensor signal model. The improvements in reliability and repeatability bring the system close to deployment for real-world trials.


Update on the Current Status and Indications for Transcatheter Edge-to-Edge Mitral Valve Repair.
To review the current status and indications of transcatheter edge-to-edge mitral valve repair. Mitral regurgitation remains a common valvular disease and can be classified as degenerative (primary) or functional (secondary). Randomized controlled trials have shown that transcatheter edge-to-edge mitral valve repair with MitraClip is successful, safe, and effective in reducing mitral regurgitation. The US Food and Drug Administration approved MitraClip in 2013 for treatment of patients with primary mitral regurgitation at prohibitive surgical risk and in 2019 for secondary mitral regurgitation. Several MitraClip generations exist (NT/R, XT/R, NTW, and XTW) with unique features and considerations. Additional edge-to-edge repair, non-edge-to-edge repair, and transcatheter valve replacement systems are under investigation as stand-alone or adjunctive therapy for patients with mitral regurgitation. Mitral regurgitation remains a significant health burden and many patients are not suitable for surgical repair or replacement. Transcatheter mitral valve therapies can be considered in selected patients and are safe and effective. More research is needed to understand how to best select devices and patients and optimize outcomes.


School dropouts: examining the space of reasons.
Dropping out of school is a worldwide phenomenon with drastic mental health consequences for children, families and society. AIM AND MATERIALS METHODS: This study examines school dropouts in one district in Kerala with an emphasis on looking at multiple reasons for the problem. The most common "reason" was various Physical disorders (80, 21.8%) followed by Mental Retardation (77, 20.9%). Child labour (Employment) came last (30, 8.1%) as a "reason" while financial issues constituted 50 (13.6%). Family issues accounted for 63 (17.1%) and School-related issues 68 (18.5%). This study highlights the need to examine a space of reasons for this phenomenon with active involvement and coordination of multiple agencies to examine and support getting children back to school and prevent dropouts.


Cherubism--an initial unilateral presentation.
An unusual presentation of cherubism is reported owing to the initial unilateral nature and late onset of occurrence. A classification for cherubism is proposed and the difficulty in diagnosis of unilateral cases is discussed.


Photo-Stacking Technique for Neuroanatomical High-Definition Photography and 3-Dimensional Modeling.
Three-dimensional (3D) neuroanatomical knowledge is vital in neurosurgery. Technological advances improved 3D anatomical perception, but they are usually expensive and not widely available. The aim of the present study was to provide a detailed description of the photo-stacking technique for high-resolution neuroanatomical photography and 3D modeling. The photo-stacking technique was described in a step-by-step approach. The time for image acquisition, file conversion, processing, and final production was measured using 2 processing methods. The total number and file size of images are presented. Measures of central tendency and dispersion report the measured values. Ten models were used in both methods achieving 20 models with high-definition images. The mean number of acquired images was 40.6 (14-67), image acquisition time 51.50 ± 18.8 s, file conversion time 250 ± 134.6 s, processing time 50.46 ± 21.46 s and 41.97 ± 20.84 s, and 3D reconstruction time was 4.29 ± 0.74 s and 3.89 ± 0.60 s for methods B and C, respectively. The mean file size of RAW files is 1010 ± 452 megabyte (MB) and 101.06 ± 38.09 MB for Joint Photographic Experts Group files after conversion. The mean size of the final image means size is 71.9 ± 0.126 MB, and the mean file size of the 3D model means is 37.4 ± 0.516 MB for both methods. The total equipment used was less expensive than other reported systems. The photo-stacking technique is a simple and inexpensive method to create 3D models and high-definition images that could prove valuable in neuroanatomy training.


Development and validation of a qualitative screening method for the detection of exogenous anabolic steroids in urine by liquid chromatography-tandem mass spectrometry.
A screening method for the urinary detection of 34 exogenous anabolic steroids has been developed. The method involves an enzymatic hydrolysis, liquid-liquid extraction and detection by liquid chromatography-tandem mass spectrometry. The use of some adducts such as [M+NH(4)](+), [M+CH(3)COO](-) and [M+H+MeOH](+) was necessary in order to detect some analytes at the required level (lower than 10 ng/ml). Two transitions were selected for each analyte. Different concentration factors have been studied in order to increase the sensitivity. A concentration factor of 50 was selected for the screening method although the high ion suppression observed under these conditions can hamper its application as a quantitative method. The method was validated and limits of detection were obtained by spiking ten different blank urine samples at five different concentration levels. Up to 29 analytes were detected in all spiked urines at the required level. Limits of detection between 1 and 10 ng/ml were obtained for most analytes which fulfil current requirements. The applicability of the method was shown by analysing positive samples.


Maximal motor unit firing rates during isometric resistance training in men.
This study measured changes in maximal voluntary contraction (MVC) force, percentage maximal activation, maximal surface EMG, M-wave amplitude and average motor unit firing rates during the initial 3 weeks of isometric resistance training of the quadriceps muscle. Ten men participated in a resistance training programme three times a week for 3 weeks and 10 men participated as a control group. In the training group, MVC increased by 35% (from 761 +/- 77 to 1031 +/- 78 N) by the end of the 3 weeks. There were no changes in mean motor unit firing rates during submaximal or maximal voluntary contractions of 50 (15.51 +/- 1.48 Hz), 75 (20.23 +/- 1.85 Hz) or 100% MVC (42.25 +/- 2.72 Hz) with isometric resistance training. There was also no change in maximal surface EMG relative to the M-wave amplitude. However, there was a small increase in maximal activation (from 95.7 +/- 1.83 to 98.44 +/- 0.66%) as measured by the twitch interpolation technique. There were no changes in any of the parameters measured in the control group. It is suggested that mechanisms other than increases in average motor unit firing rates contributed to the increase in maximal force output with resistance training. Such mechanisms may include a combination of increased motor unit recruitment, enhanced protein synthesis, and changes in motor unit synchronization and muscle activation patterns across the quadriceps synergy.


Machine learning and statistical models for analyzing multilevel patent data.
A recent surge of patent applications among public hospitals in China has aroused significant research interest. A country's healthcare innovation capacity can be measured by its number of patents. This paper explores the link between the number of patents and ten independent variables. Multicollinearity was carefully detected and removed by using the variable selection method and LASSO regression, respectively. The Poisson model and the negative binomial model were proposed to analyze the patent data. Three goodness of fit tests, the Pearson test, the deviance test, and the DHARMa non-parametric dispersion test, were conducted to investigate if the model has a good fit. After discovering four clusters by conducting agglomerative hierarchical clustering, these two models were replaced by the negative binomial mixed model. The likelihood ratio test was used to determine which model is more appropriate and the results reveal that the negative binomial mixed model outperforms both the Poisson model and the negative binomial model. Three variables, number of health technicians per 10,000 people, financial expenditure on science and technology as well as number of patent applications per 10,000 health personnel, have a significantly positive relationship with the number of patents in Chinese tertiary public hospitals.


A case report of gastrointestinal histoplasmosis in a patient treated with infliximab.
Histoplasmosis is an endemic mycosis in some areas of North and South America. This disease is usually asymptomatic, but it can result in severe and disseminated infection involving gastrointestinal tract, especially in immunocompromised individuals. We report a case of a 33-years-old Ecuadorian male treated with infliximab who developed disseminated histoplasmosis with gastrointestinal affection. Due to the non-specific presentation of gastrointestinal histoplasmosis, the diagnosis is often delayed and it causes poor outcomes. It is important to consider this diagnosis in immunocompromised patients with compatible symptoms, like patients on TNF inhibitors.


Hypoalbuminemia Increases Mortality after Two-Stage Revision Total Joint Arthroplasty.
This study aimed to evaluate the effect of hypoalbuminemia on failure rates and mortality after a two-stage revision for PJI. 199 Patients (130 knees and 69 hips) with a mean age of 64.7 ± 10.7 years who underwent a two-stage exchange were retrospectively reviewed at a mean of 51.2 ± 39.7 months. Failure of treatment was defined as any revision within the follow-up period, failure to undergo reimplantation, or death within one year of initiating treatment. There were 71 failures (35.7%), including 38 septic failures (19.1%). We found no differences between successful revisions and failures regarding hypoalbuminemia (43% vs. 42% prior to stage 1, P=1 and 32% vs. 29% prior to stage 2, P=0.856). There were also no differences in hypoalbuminemia rates between septic failures and the rest of the cohort (42% vs. 43% prior to stage 1, P=1.0 and 34% vs. 30% prior to stage 2, P=0.674). Hypoalbuminemia prior to stage 2 was a significant predictor of mortality based on multivariate analysis (odds ratio 5.40, CI 1.19-24.54, P=0.029). Hypoalbuminemia was independently associated with a greater length of stay by 2.2 days after stage 1 (P=0.002) and by 1.0 days after the second stage reimplantation (P=0.004). Preoperative hypoalbuminemia is a significant predictor of mortality and increased length of stay following two-stage revision but is not a predictor of failure of PJI treatment. Further study is required to understand if hypoalbuminemia is a modifiable risk factor or a marker for poor outcomes.


Absence of orgasm-induced prolactin secretion in a healthy multi-orgasmic male subject.
In several studies we have recently demonstrated that orgasm induces prolactin secretion in healthy males and females. This suggests that prolactin may form a feedback regulator of the refractory period following orgasm. To examine this position we investigated the prolactin response of a healthy multi-orgasmic male subject. Blood was drawn continuously during masturbation-induced orgasm. The prolactin response of the case-subject was compared with that of nine healthy adult men with a normal refractory period. The case-subject showed no prolactin response to three orgasms. Data from this multi-orgasmic subject support the hypothesized role of plasma prolactin in contributing to sexual-satiation mechanisms.


Receptor pharmacophore ensemble (REPHARMBLE): a probabilistic pharmacophore modeling approach using multiple protein-ligand complexes.
Ensemble methods are gaining more importance in structure-based approaches as single protein-ligand complexes strongly influence the outcomes of virtual screening. Structure-based pharmacophore modeling based on a single protein-ligand complex with complex feature combinations is often limited to certain chemical classes. The REPHARMBLE (receptor pharmacophore ensemble) approach presented here examines the ability of an ensemble of selected protein-ligand complexes to populate pharmacophore space in the ligand binding site, rigorously assesses the importance of pharmacophore features using Poisson statistic and information theory-based entropy calculations, and generates pharmacophore models with high probabilities. In addition, an ensemble scoring function that combines all the resultant high-scoring pharmacophore models to score molecules is derived. The REPHARMBLE approach was evaluated on ten DUD-E benchmark datasets and afforded good screening performance, as measured by receiver operating characteristic, enrichment factor and Güner-Henry score. Although one of the high-scoring models achieved superior statistical results in each dataset, the ensemble scoring function balanced the shortcomings of each model and passed with close performance measures. This approach offers a reliable way of choosing the best-scoring features to build four-feature pharmacophore queries and customize a target-biased 'pharmacophore ensemble' scoring function for subsequent virtual screening.


Filtration surgery in the treatment of neovascular glaucoma.
The disappointing visual acuity results following cyclocryotherapy for neovascular glaucoma have prompted us to consider filtration surgery as a reasonable alternative in the treatment of selected patients. All cases of surgically treated neovascular glaucoma were reviewed. There were 26 operations in 24 eyes with a follow-up ranging from six months to seven years and a mean of 22.8 months. Adequate pressure control was obtained in 16 of the 24 eyes (67%). In the eyes with successful control of intraocular pressure, vision was 20/400 or better in eight of the 16 eyes (50%). Four patients (17%) lost light perception. Approximately one half of the operations were trabeculectomies, and the others were posterior lip sclerectomies. There seemed to be no significant difference in the final intraocular pressure levels, number or degree of complications, or successes within the two subgroups. Panretinal photocoagulation, topical steroids, cycloplegics, and time for these measures to have their effect are important preoperative adjuncts to surgical treatment. We are encouraged by the long-term preservation of vision and control of intraocular pressure in many of these eyes.


Cranioplasty: indications, technique, and early results of autogenous split skull cranial vault reconstruction.
Since 1990, fresh autogenous split skull cranioplasty has been the preferred method of reconstructing complicated cranial defects at the Sunnybrook Hospital. Eight cases of split skull cranioplasties were reviewed to illustrate the indications and technical considerations of the procedure. Clinical factors believed to be detrimental to the outcome of split skull cranioplasty were identified by: 1) a history of an infected, failed, previous cranial reconstruction, 2) inadequate scalp or skin to cover the external surface of an anticipated skeletal vault reconstruction, 3) the communication of adjacent paranasal sinuses with the epidural space, 4) the presence of an avascular dead space posterior to the reconstructed cranium. These potentially negative influences were systematically addressed as part of the surgical routine in autogenous split skull cranioplasty.


Preoperative CT Radiomics Nomogram for Predicting Microvascular Invasion in Stage I Non-Small Cell Lung Cancer.
RATIONALE AND OBJECTIVES: This study aims to develop and validate a nomogram integrating clinical-CT and radiomic features for preoperative prediction of microvascular invasion (MVI) in patients with stage I non‑small cell lung cancer (NSCLC). This retrospective study analyzed 188 cases of stage I NSCLC (63 MVI positives and 125 negatives), which were randomly assigned to training (n = 133) and validation cohorts (n = 55) at a ratio of 7:3. Preoperative non-contrast and contrast-enhanced CT (CECT) images were used to analyze computed tomography (CT) features and extract radiomics features. The student's t-test, the Mann-Whitney-U test, the Pearson correlation, the least absolute shrinkage and selection operator, and multivariable logistic analysis were used to select the significant CT and radiomics features. Multivariable logistic regression analysis was performed to build the clinical-CT, radiomics, and integrated models. The predictive performances were evaluated through the receiver operating characteristic curve and compared with the DeLong test. The integrated nomogram was analyzed regarding discrimination, calibration, and clinical significance. The rad-score was developed with one shape and four textural features. The integrated nomogram incorporating radiomics score, spiculation, and the number of tumor-related vessels (TVN) demonstrated better predictive efficacy than the radiomics and clinical-CT models in the training cohort (area under the curve [AUC], 0.893 vs 0.853 and 0.828, and p = 0.043 and 0.027, respectively) and validation cohort (AUC, 0.887 vs 0.878 and 0.786, and p = 0.761 and 0.043, respectively). The nomogram also demonstrated good calibration and clinical usefulness. The radiomics nomogram integrating the radiomics with clinical-CT features demonstrated good performance in predicting MVI status in stage I NSCLC. The nomogram may be a useful tool for physicians in improving personalized management of stage I NSCLC.


Lobar bronchial atresia demonstrating a cystic lesion without overinflation.
Congenital bronchial atresia (CBA) is an infrequent pulmonary anomaly characterized, in general, by a blindly terminating bronchus, mucoid impaction, and hyperinflation of the peripheral pulmonary parenchyma. We herein report an adult case of lobar bronchial atresia who showed no hyperinflation of the peripheral pulmonary parenchyma.


A Randomized Trial of Clopidogrel vs Ticagrelor After Off-Pump Coronary Bypass.
This study aimed to compare the outcomes of aspirin in combination with either ticagrelor or clopidogrel after off-pump coronary artery bypass (OPCAB) in patients with clopidogrel resistance. Between November 2014 and November 2020, 1739 patients underwent OPCAB. Aspirin and clopidogrel treatment was initiated the day after surgery. On postoperative days 7 to 9, clopidogrel resistance was evaluated using a point-of-care assay. A total of 278 (18.9%) patients had clopidogrel resistance ( platelet reaction unit >208) and were enrolled in the study. The study investigators excluded patients with coresistance to aspirin (n = 74) and divided the remaining patients (mean age, 67.4 ± 8.5 years) into 2 groups (an aspirin and ticagrelor group [AT group; n = 102] and an aspirin and clopidogrel group [AC group; n = 102]), randomly assigned using a 1:1 ratio block table. The primary end point was graft patency and major adverse cardiovascular events (MACEs; defined as the composite of cardiovascular mortality, myocardial infarction, and repeat revascularization at 1 year after OPCAB), and the coprimary end point was the graft patency rate. The data were analyzed using the intent-to-treat method. The graft occlusion rates in the AT and AC groups were 3.9% and 5.9%, respectively (P = .52). Neither death from cardiovascular causes (1.0% vs 2.9%; P = .32) nor myocardial infarction showed significant differences (1.0% vs 3.9%; P = .18). No significant difference in the rates of major bleeding were found between the 2 groups (P = .75). However, the AT group was associated with a lower rate of MACEs after OPCAB (hazard ratio, 0.77; 95% CI, 0.684-0.891; P = .01). These results suggest that ticagrelor may be associated with reducing MACEs in patients with clopidogrel resistance after OPCAB.


Fine-needle aspiration cytology of non-neoplastic adrenal pathology.
The incidence of many fungal and parasitic lesions are on the rise over the last few decades. In this study, we have analyzed the spectrum of non-malignant lesions in adrenal gland diagnosed on fine-needle aspiration biopsy (FNAB) in a tertiary health care center. FNAB of the adrenal gland was done under CT (4 cases) and ultrasound guidance (11 cases). In six cases, the lesions were present in both the adrenal glands. Special stains were done whenever needed. No complications were encountered in any of the FNAB procedure. Detailed cytomorphological study was done along with clinical history. There were total of 15 cases comprising of 5 tuberculosis, 5 Histoplasma, 2 acute inflammation, 2 granulomatous inflammation, and 1 myelolipoma. Acid fast bacilli positive on Ziehl Neelsen stain were labeled as mycobacterial infection possibly tuberculosis. The capsule of Histoplasma was bright pink on Periodic Acid Schiff stain. FNAB is an easy, reliable, and minimally invasive method to diagnose and categorize the various benign non-neoplastic diseases of adrenal gland. Diagn. Cytopathol. 2016;44:472-476. © 2016 Wiley Periodicals, Inc.


BASIC: A Simple and Accurate Modular DNA Assembly Method.
Biopart Assembly Standard for Idempotent Cloning (BASIC) is a simple, robust, and highly accurate DNA assembly method, which provides 99% correct assemblies for a typical four-part assembly, enabling high efficiency cloning workflows (Storch et al., ACS Synth Biol, https://doi.org/10.1021/sb500356 , 2015). BASIC employs standardised DNA linkers to combine bioparts, stored in the universal BASIC format. Once a new biopart is formatted into BASIC standard, defined by flanking 18 bp prefix and suffix sequences, it can be placed at any position and in any context within a designed BASIC assembly. This modularity of the BASIC approach is further enhanced by a range of functional linkers, including genetic elements like ribosomal binding sites (RBS) and peptide linkers. The method has a single tier format, whereby any BASIC assembly can create a new composite BASIC part in the same format used for the original parts; it can thus enter a subsequent BASIC assembly without the need for reformatting or changes to the workflow. This unique idempotent cloning mechanism allows for the assembly of constructs in multiple, conceptionally simple hierarchical rounds. Combined with its high accuracy and robustness, this makes BASIC a versatile assembly method for combinatorial and complex assemblies both at bench and biofoundry scale. The single universal storage format of BASIC parts enables compressed universal biopart libraries that promote sharing of parts and reproducible assembly strategies across labs, supporting efforts to improve reproducibility. In comparison with other DNA assembly standards and methods, BASIC offers a simple robust protocol, relies on a single tier format, provides for easy hierarchical assembly, and is highly accurate for up to seven parts per assembly round (Casini et al., Nat Rev Mol Cell Biol. https://doi.org/10.1038/nrm4014 , 2015).


Production of polyunsaturated fatty acids by Schizochytrium (Aurantiochytrium) spp.
Diverse health benefits are associated with dietary consumption of omega-3 long-chain polyunsaturated fatty acids (ω-3 LC-PUFA), particularly docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Traditionally, these fatty acids have been obtained from fish oil, but limited supply, variably quality, and an inability to sustainably increase production for a rapidly growing market, are driving the quest for alternative sources. DHA derived from certain marine protists (heterotrophic thraustochytrids) already has an established history of commercial production for high-value dietary use, but is too expensive for use in aquaculture feeds, a much larger potential market for ω-3 LC-PUFA. Sustainable expansion of aquaculture is prevented by its current dependence on wild-caught fish oil as the source of ω-3 LC-PUFA nutrients required in the diet of aquacultured animals. Although several thraustochytrids have been shown to produce DHA and EPA, there is a particular interest in Schizochytrium spp. (now Aurantiochytrium spp.), as some of the better producers. The need for larger scale production has resulted in development of many strategies for improving productivity and production economics of ω-3 PUFA in Schizochytrium spp. Developments in fermentation technology and metabolic engineering for enhancing LC-PUFA production in Schizochytrium spp. are reviewed.


Pharmacological characterization of histamine H2 receptors on clonal cytolytic T lymphocytes. Evidence for histamine-induced desensitization.
Cultured cytolytic T lymphocytes of clonal origin were screened for histamine-stimulated cyclic AMP production. Histamine caused a 2- to 8-fold elevation of cyclic AMP levels in five independent clones. The EC50 for histamine of 1.7 X 10(-5) M and the rank order of potencies of H1 and H2 agonists [impromidine greater than histamine greater than dimaprit greater than 4-methylhistamine greater than 2-methylhistamine greater than 2-(2-aminoethyl)-thiazole] were characteristic of the conventional histamine H2 receptor. H1 and H2 antagonists inhibited histamine-stimulated cyclic AMP elevation with inhibition constants typical for those found on other H2 receptor systems. Prior incubation of cells with histamine resulted in a marked loss in responsiveness to subsequent histamine challenge. We demonstrate that this desensitization is dose and time dependent and results in a change in the efficacy and not the potency of histamine. Although cyclic AMP increases could also be elicited with isoproterenol, prostaglandin E1 or forskolin, desensitization of histamine had no effect on the ability of these agents to stimulate cyclic AMP production. In contrast to the rapid rate of histamine-induced desensitization, recovery of histamine responsiveness could not be detected for several hours.


Carbon dot based sensing platform for real-time imaging Cu2+ distribution in plants and environment.
Divalent copper is a double-edged sword for plants, excess or shortage of copper ions will cause adverse reactions in plants. Currently, Cu2+ sensor for plants is still underdeveloped and new technology is urgently required for realizing one-step and real-time detection of Cu2+ in plants. Herein, a home-made and low-cost sensing platform is constructed by using carbon dots (CDs) as the optical probe, electronic devices for image acquisition, and a built-in algorithm program for image processing, which allows the dynamic monitoring of Cu2+ distribution in different plant species with high spatial and temporal resolution. We found that the detection limit of R-CDs for Cu2+ in water sample was 0.375 nM, and 11.7 mg/kg or even less Cu2+ in plants can be visually observed and accurately detected by the sensing platform. Moreover, this sensing platform has also been employed for reporting the spatial distribution of Cu2+ in the external environment of plants, demonstrating its applicability for monitoring Cu2+ both in living plants and the surrounding environment. This study provides a smart sensing platform for precise detection in plant internal and external environments, offering a promising strategy for precision agriculture in real-time and remote-control manners.


Interface haze associated with topical nonsteroidal anti-inflammatory drug use after descemet stripping automated endothelial keratoplasty.
To determine if topical nonsteroidal anti-inflammatory drug (NSAID) use after Descemet stripping automated endothelial keratoplasty (DSAEK) is associated with the development of interface haze. A retrospective case-control study of patients undergoing DSAEK surgery. Of the 61 patients receiving topical NSAIDs, 51% were noted to develop interface haze, and 9% of the control group (N=100, no NSAID) developed haze. There was no significant difference in best corrected visual acuity between the 2 groups. Topical NSAID use is associated with the development of interface haze, although it may not be visually significant and could be related to confounding factors.


Discrimination of botanical origin of olive oil from selected Greek cultivars by SPME-GC-MS and ATR-FTIR spectroscopy combined with chemometrics.
Consumers today wish to know the botanical origin of the olive oil they purchase. The objective of the present study was the development of robust chemometric models based on gas chromatography-mass spectrometry (GC-MS) and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) for the purpose of botanical differentiation of three commercial Greek olive oil cultivars. Using the solid-phase microextraction technique (SPME), volatile compounds (VC) were obtained and analyzed by GC-MS. Five hydrocarbons and one ester were selected by the forward stepwise algorithm, which best discriminated the olive oil samples. From ATR-FTIR analysis, the spectral regions chosen from the forward stepwise algorithm were associated with CO stretching vibration of the esters of triglycerides and the CH bending vibrations of the CH2 aliphatic group and double bonds. Application of the supervised methods of linear and quadratic discriminant cross-validation analysis, based on VC data, provided a correct classification score of 97.4% and 100.0%, respectively. Corresponding statistical analyses were used in the mid-infrared spectra, by which 96.1% of samples were discriminated correctly. ATR-FTIR and SPME-GC-MS techniques in conjunction with the appropriate feature selection algorithm and classification methods proved to be powerful tools for the authentication of Greek olive oil. The proposed methodology could be used in an industrial setting for determination of the botanical origin of Greek olive oil. © 2020 Society of Chemical Industry.


Sensitivity to inhibitors of type II topoisomerases from mouse L5178Y lymphoma strains that are resistant or sensitive to X-radiation.
Sensitivity to topoisomerase II inhibitors was tested at the cellular and enzyme level for two strains of mouse L5178Y lymphoma cells: resistant (LY-R) and sensitive (LY-S) to X-radiation. Differences in the susceptibility to inhibitors between LY-R and LY-S cells depended on the inhibitor used and were observed for adriamycin and VP-16, but not for mitoxantrone. On the other hand, isolated enzymes displayed the same sensitivity to all inhibitors tested regardless of the cell line. These results exclude the presence of altered topoisomerase II in LY-S cells as a possible reason for the collateral sensitivity of LY-S cells to X-radiation and topoisomerase II inhibitors.


Flecainide therapy suppresses exercise-induced ventricular arrhythmias in patients with CASQ2-associated catecholaminergic polymorphic ventricular tachycardia.
Calsequestrin-associated catecholaminergic polymorphic ventricular tachycardia (CPVT2) can cause sudden death in young individuals in response to stress. Beta-blockers are the mainstay medical treatment for patients with CPVT2. However, they do not prevent syncope and sudden death in all patients. Flecainide was reported to reduce exercise-induced ventricular arrhythmias (EIVA) in patients with ryanodine receptor-associated CPVT. The role of flecainide in CPVT2 is not known. To summarize our experience in combining flecainide and beta-blockers in high-risk patients with CPVT2. All patients with CPVT2 (10 patients) who have high-risk features (syncope, EIVA, or appropriate implantable cardioverter-defibrillator [ICD] shocks) despite beta-blockers with or without calcium channel blockers were treated with a combination of flecainide and beta-blockers. Exercise test was done before and after beginning treatment with flecainide. All patients had EIVA and 4 had appropriate ICD shocks before flecainide treatment. EIVA-included frequent ventricular premature beats and or ventricular tachycardia during the exercise test while on high dose of beta-blockers with or without calcium channel blockers before treatment with flecainide. After combination therapy with flecainide and beta-blockers, EIVA were suppressed completely in all patients. During follow-up of 15.5 ± 10.4 months (range 2-29 months), 8 patients were free of symptoms and free of arrhythmias. Two patients had 1 VT storm episode with recurrent ICD shocks despite repeated normal stress test. Flecainide can completely prevent ventricular arrhythmia during exercise and partially prevent recurrent ICD shocks in high-risk patients with CPVT2.


[Use of computer-assisted stabilography and the method of computer-assisted biomechanical study of gait in the diagnosis of posture and motor disorders in patients with different forms of infantile cerebral paralysis].
The paper presents and analyzes the results of assessment of vertical stability and walk in patients with infantile cerebral paralysis before and after dynamic proprioceptive correction, which is of great importance for differential diagnosis of different forms of this disease, for controlling the course of therapeutical measures, for working out criteria for the efficiency of treatment.


[Analyze and compare activities of polysaccharide form rhizoma chuanxiong and radix paeoniae rubra].
To explore the difference of biological activities between two Chinese Herbs polysaccharides. Extracted two kinds of Chinese Herbs Polysaccharides by hot water, and determined the content of Polysaccharides. Determined the antibacterial activity with turbidimetry. Explored antioxidant activity using an in vitro antioxidant analysis, probed into the effects on cell growth with MTT colorimetric analysis. The two polysaccharides both showed activities on the antibacterial, anti-oxidation and promotion of cell growth but different in degree. The Radix Paeoniae Rubra polysaccharide was stronger than Rhizoma Chuanxiong in antioxidant and antimicrobial activities, and weaker in promotion of cell growth activity. Activity differences exist between the two polysaccharides and the activities show some complementary effects. The differences provide important value in research on polysaccharide compound.


Low expression of asparagine synthetase in lymphoid blasts precludes its role in sensitivity to L-asparaginase.
Childhood acute lymphoblastic leukemia (ALL) is treated with combined chemotherapy, including L-asparaginase (L-asp). Recent studies question the traditional view that the level of asparagine synthetase (ASNS), an enzyme producing the intracellular asparagine, correlates with the response to L-asp treatment. However, the importance of ASNS in response to L-asp has neither been confirmed nor refuted so far. In this study, we wanted to elucidate the effect of ASNS expression level on the sensitivity of ALL cells to L-asp treatment. We used four ALL cell lines (NALM-6, RS4;11, REH, and UOCB6) and 30 diagnostic bone marrow samples of ALL patients to study the relationship between ASNS expression and sensitivity to L-asp using MTS proliferation assay. RNA interference was used to study the effect of a range of ASNS levels on the response to L-asp treatment. Using a cell line model with a gradually knocked-down ASNS gene, we defined a cutoff level below which ASNS gene expression does not correlate with sensitivity to L-asp. Importantly, ASNS gene expression in patients' ALL blasts is below this level. We confirmed that there was no correlation between ASNS gene expression and sensitivity to L-asp in ALL blasts. In addition, we show that cells with low ASNS expression level do not respond to asparagine deprivation by upregulation of ASNS gene expression. In conclusion, the ASNS expression level does not predict sensitivity to L-asp in leukemic blasts. Moreover, cell lines with high basal expression of ASNS cannot serve as a valid model for studies on the relationship between the ASNS and L-asp cytotoxic effect.


Validity evidence for a virtual multiple mini interview at a pharmacy program.
Numerous health professions schools have transitioned to virtual admissions interviews in recent years. While some research suggests that virtual multiple mini-interviews (vMMIs) are feasible, acceptable, and more affordable, there is a paucity of research concerning the validity of this approach. The purpose of this study was to examine the validity and reliability of vMMIs and explore differences in performance between vMMI and in-person MMIs. Data were collected for two years of in-person MMIs and two years of vMMIs at a pharmacy program/school in the United States. An exploratory factor analysis (principal components analysis) with varimax rotation and Kaiser rule (i.e. retaining factors with eigenvalue > 1.0) was used to explore the construct validity of the vMMI data. Pearson correlation was used to examine correlations between vMMI stations and Cronbach alpha was used to determine the internal consistency of each station. Independent t-tests were used to examine differences between in-person MMI and vMMI scores. Cohen's d was used to determine effect sizes. Four hundred and thirty-eight (42.69%) candidates completed an in-person MMI and 588 (57.31%) completed a vMMI. Factor analysis indicated that each vMMI station formed a single factor with loads ranging from 0.86 to 0.96. The vMMI stations accounted for most of the total variance, demonstrated weak to negligible intercorrelations, and high internal consistency. Significant differences between in-person and vMMI scores were found for the teamwork-giving, teamwork-receiving, and integrity stations. Medium effect sizes were found for teamwork-giving and teamwork-receiving and a small effect size was found for integrity. Initial evidence suggests that the vMMI is a valid and reliable alternative to in-person MMIs. Additional research is needed to examine sources of differences in rating patterns between the two approaches and identify strategies that align with institutional priorities for recruitment and admissions.


[Effect of stimulation of alpha and beta presynaptic adrenoceptors on the liberation of noradrenaline produced by electric stimulation in rabbit carotid sinus].
The experiments carried out on the isolated carotid sinus in the rabbit demonstrated that alpha and beta adrenoceptors are capable of detecting the circulating catecholamines and the catecholamines of the synaptic cleft in order to modulate the release of noradrenaline from the sympathetic nerve endings in response to a potential action.


Mannose-binding lectin polymorphisms and rheumatoid arthritis: A short review and meta-analysis.
Mannose-binding lectin (MBL) is a pattern recognition receptor of the lectin pathway of complement system. MBL binds to carbohydrates on microorganism's surfaces leading to complement activation, opsonization and phagocytosis. Polymorphisms in the MBL gene (MBL2) are associated with variations on MBL serum levels and with the susceptibility to various infectious and autoimmune diseases. The involvement of the lectin pathway in rheumatoid arthritis (RA) has been demonstrated by several studies and although MBL has been considered to have a dual role in the pathogenesis of the disease, the association between MBL and RA remains inconclusive. In an attempt to clarify this relationship, we developed this short review summarizing accumulated evidences in regard to MBL and RA and a meta-analysis to evaluate the influence of MBL2 polymorphisms on the susceptibility to RA. Among a total of 217 articles that were identified following a predefined search strategy on PubMed, Scopus, Scielo, EMBASE and Cochrane databases, only 13 met all inclusion criteria and were included in the meta-analysis. Data assessment was conducted by three independent investigators and presented in odds ratio (OR) and 95% confidence intervals (CIs) using forest plot charts. Both heterogeneity and publication bias were analyzed. The results of the meta-analysis evidenced that MBL2 low producing OO and XX genotypes do not confer higher risk to RA, even when data were analyzed according to cohort's ethnicity. Further studies are needed in order to clarify the importance of other genes of the lectin pathway in the pathogenesis of RA.


Does preoperative pharmacologic prophylaxis reduce the rate of venous thromboembolism in pancreatectomy patients?
Whether the risk of venous thromboembolism (VTE) may be reduced by preoperative administration of prophylactic heparin is unknown. We hypothesized that timing of heparin administration does not significantly alter the incidence of VTE in pancreatic surgery. An analysis was conducted using data from Massachusetts General Hospital's National Surgical Quality Improvement Program from 2012 to 2017. All patients admitted for elective pancreatic resection were included. The primary outcome was development of VTE. Multivariable regression was performed, adjusting for patient demographics and various clinical factors. In total, 1448 patients were analyzed, of whom 1062 received preoperative heparin (73.3%). Overall, 36 (2.5%) patients developed VTE. On unadjusted analysis, there was no statistically significant difference between patients who received preoperative heparin compared with those who did not (2.6% vs. 1.3%, respectively; p = 0.079). On adjusted analysis, there was an association with increased VTE rates among patients who received preoperative heparin (OR 2.93, 95% CI 1.10-7.81; p = 0.031). There was an association between preoperative heparin administration and increased incidence of VTE on adjusted analysis, possibly reflecting appropriate surgical judgment in patient selection for prophylaxis. These data question the inclusion of preoperative VTE pharmacologic prophylaxis as a reliable quality indicator.


Keep in touch: the soil-root hydraulic continuum and its role in drought resistance in crops.
Drought is a major threat to food security worldwide. Recently, the root-soil interface has emerged as a major site of hydraulic resistance during water stress. Here, we review the impact of soil drying on whole plant hydraulics and discuss mechanisms by which plants can adapt by modifying the properties of the rhizosphere either directly or through interactions with the soil microbiome.


Human non-germinal center B cell interleukin (IL)-12 production is primarily regulated by T cell signals CD40 ligand, interferon gamma, and IL-10: role of B cells in the maintenance of T cell responses.
Interleukin (IL)-12 is expressed mainly in antigen-presenting cells after challenge with microbial material or after CD40 activation. Although IL-12 was cloned from human Epstein-Barr virus (EBV)-transformed B cell lines, surprisingly, CD40 ligation on murine B cells did not lead to IL-12 production, suggesting that murine B cells do not produce IL-12. Here we demonstrate that a subset of human tonsillar B cells can be induced to express and secrete bioactive IL-12. The major stimulus to produce IL-12 in human B cells was CD40 ligation. In contrast, B cell receptor cross-linking did not induce IL-12. Expression of IL-12 after CD40 activation was restricted to CD38(-)IgD+/- non-germinal center (non-GC) B cells. CD40 ligation and interferon (IFN)-gamma exhibited synergistic effects on IL-12 production, whereas IL-10 abrogated and IL-4 significantly inhibited IL-12 production by these B cells. In contrast to IL-12, production of IL-6 is conversely regulated, leading to significant increase after CD40 ligation in the presence of the T helper type 2 (Th2) cytokine IL-4. Cord blood T cells skewed towards either a Th1 or a Th2 phenotype maintained their cytokine expression pattern when restimulated with allogeneic resting B cells. Blockade of CD40 and/or IL-12 during T-B interaction significantly reduced IFN-gamma production by the T cells. This suggests a model whereby B cells produce either IL-12 or IL-6 after contact with T cells previously differentiated towards Th1 or Th2. Furthermore, IL-12 and IL-6 might provide a positive feedback during cognate T-B interactions, thereby maintaining T cells' differentiation pattern during amplification of the immune response.


Multidrug encapsulation by coaxial tri-capillary electrospray.
This study describes how a coaxial tri-capillary electrospray (ES) system can synthesize monodisperse PLGA-coated particles containing multiple drugs in one step. The coaxial arrangement of three separate capillaries in an ES nozzle enables production of composite particles with tri-layered structures without the complicated steps involved in emulsion processes. Various materials can be encapsulated in separate layers of individual composite particles without regard for the hydroscopic property of drugs. At the proper spray setting, nearly 100% drug encapsulation efficiency can be achieved. By varying the feed flowrate of spray solutions, the overall particle sizes, ranging from submicrometer to micrometer, and the thickness of the layers in produced particles can also be controlled. Size and tri-layered structure of the composite particles were characterized by SEM and laser scanning confocal microscopy (LSCM). We further explored the spray technique in the production of tri-layered composite particles having a controlled multidrug-release profile, and compared the release profiles of both core-shell structured particles (produced by coaxial dual-capillary ES) and tri-layered ones (produced by coaxial tri-capillary ES). The comparison showed that composite particles with the tri-layered structure can release multiple drugs in distinct kinetic phases. We further demonstrated that the release profile of tri-layered composite particles can be effectively controlled by varying the thickness and chemical composition of their individual layers.


Delivery of the TLR ligand poly(I:C) to liver cells in vitro and in vivo by calcium phosphate nanoparticles leads to a pronounced immunostimulation.
The selective activation of the immune system is a concurrent problem in the treatment of persistent diseases like viral infections (e.g. hepatitis). For the delivery of the toll-like receptor ligand poly(I:C), an immunostimulatory action was discovered earlier by hydrodynamic injection. However, this technique is not clinically transferable to human patients. A modular system where the immunoactive toll-like-receptor ligand 3 (TLR-3) poly(I:C) was incorporated into calcium phosphate nanoparticles was developed. The nanoparticles had a hydrodynamic diameter of 275nm and a zeta potential of +20mV, measured by dynamic light scattering. The diameter of the solid core was 120nm by scanning electron microscopy. In vitro, the nanoparticle uptake was investigated after 1 and 24h of incubation of THP-1 cells (macrophages) with nanoparticles by fluorescence microscopy. After intravenous injection into BALB/c and C57BL/6J mice, respectively, the in vivo uptake was especially prominent in lung and liver, 1 and 3h after the injection. Pronounced immunostimulatory effects of the nanoparticles were found in vitro with primary liver cells, i.e. Kupffer cells (KC) and liver sinusoidal endothelial cells (LSEC) from wild-type C57BL/6J mice. Thus, they represent a suitable alternative to hydrodynamic injection treatments for future vaccination concepts. The selective activation of the immune system is a concurrent problem in the treatment of persistent diseases like viral infections (e.g. hepatitis). For the delivery of the toll-like receptor ligand poly(I:C), an immunostimulatory action has been discovered earlier by hydrodynamic injection. However, this technique is not clinically transferable to human patients. We have developed a modular system where poly(I:C) was incorporated into calcium phosphate nanoparticles. The uptake into relevant liver cells was studied both in vitro and in vivo. After intravenous injection into mice, the in vivo uptake was especially prominent in lung and liver, 1 and 3h after the injection. The corresponding strong immune reaction proves their high potential to turn up the immune system, e.g. against viral infections, without adverse side reactions.


Bypassing Glutamic Acid Decarboxylase 1 (Gad1) Induced Craniofacial Defects with a Photoactivatable Translation Blocker Morpholino.
γ-Amino butyric acid (GABA) mediated signaling is critical in the central and enteric nervous systems, pancreas, lungs, and other tissues. It is associated with many neurological disorders and craniofacial development. Glutamic acid decarboxylase (GAD) synthesizes GABA from glutamate, and knockdown of the gad1 gene results in craniofacial defects that are lethal in zebrafish. To bypass this and enable observation of the neurological defects resulting from knocking down gad1 expression, a photoactivatable morpholino oligonucleotide (MO) against gad1 was prepared by cyclization with a photocleavable linker rendering the MO inactive. The cyclized MO was stable in the dark and toward degradative enzymes and was completely linearized upon brief exposure to 405 nm light. In the course of investigating the function of the ccMOs in zebrafish, we discovered that zebrafish possess paralogous gad1 genes, gad1a and gad1b. A gad1b MO injected at the 1-4 cell stage caused severe morphological defects in head development, which could be bypassed, enabling the fish to develop normally, if the fish were injected with a photoactivatable, cyclized gad1b MO and grown in the dark. At 1 day post fertilization (dpf), light activation of the gad1b MO followed by observation at 3 and 7 dpf led to increased and abnormal electrophysiological brain activity compared to wild type animals. The photocleavable linker can be used to cyclize and inactivate any MO, and represents a general strategy to parse the function of developmentally important genes in a spatiotemporal manner.


Circulating salivary and serum miRNA-182, 320a, 375 and 503 expression levels in type 2 diabetes.
Early-stage diagnosis of diabetes through non-invasive and diagnostic biofluid-like saliva has become a very popular approach to facilitate future preventive interventions and improve patient care. Meanwhile, the alteration of small non-coding RNA in human fluids has been suggested as a probable precedent for the early stages of diabetes. In the present study, we checked the expression of miR-320a, 182-5p, 503, and 375 by using quantitative PCR in both stimulated and unstimulated saliva and blood samples of 40 adult patients with type-2 diabetes compared to 40 healthy individuals. In addition, we have sought to understand the possibility that miRNAs could provide new information about the status of type 2 diabetes in salivary samples beyond what can now be identified from blood samples and link their expression to the presence of clinically relevant risk factors. For this purpose, we have used a set of multivariate models. The results showed that three miRNAs were more highly expressed in patients with type 2 diabetes, while miR-320-a was down-regulated in those patients compared to healthy subjects. Furthermore, the data showed that miR-320a was the most reliable predictor for distinguishing diabetic patients from healthy subjects, with AUCs of 0.997, 0.97, and 0.99 (97.4% sensitivity and 100% specificity, p = 0.001) for serum, unstimulated, and stimulated saliva samples, respectively. Interestingly, the results of this study indicated that the amount of four miRNAs expressed in stimulated saliva was the same as in serum samples, which could conclude that specific miR-320a and 503 in stimulated saliva may introduce credible, non-invasive, and diagnostic biomarkers that can be used to monitor diabetic patients' status, while there is a need to design more research studies to confirm these findings.


Role of the amine and phosphine groups in oleylamine and trioctylphosphine in the synthesis of copper chalcogenide nanoparticles.
The effect of the functional groups of capping agents was investigated in the synthesis of copper selenide, copper sulphide and copper oxide nanoparticles using oleylamine (OLA) and trioctylphosphine (TOP). These capping molecules have demonstrated their ability to act as reducing agents, surfactants, solvents and enhancement of colloidal stabilization. They also offer electron donating abilities from the two group 5A elements, P and N. Nitrogen atom in an amine group possess stronger surface interactions and higher basicity than P atom in the phosphines. Copper chalcogenide nanoparticles were prepared using Hot-injection method and characterized using UV/Vis spectroscopy, TEM and XRD. The optical and structural properties of the yielded nanoparticles showed dependence on the type of capping interactions from the two agents. Nanoparticles synthesized using TOP produced two phases whereas a single phase was observed from OLA as confirmed by XRD. Although TOP and OLA exhibit similar features, but their affinity to metals differs resulting to significant different morphology and crystallinity of the produced nanoparticles. Amine group has higher affinity for protons than phosphine due to the lone pair of electrons it possesses which it easily donates to H+ compared to phosphine. The high proton affinity of oleylamine makes it interact faster than trioctylphosphine. OLA in overall produced larger particle sizes compared to TOP but generated a wider variety of shapes.


Neuronal network-based biomimetic chip for long-term detection of olfactory dysfunction model in early-stage Alzheimer's disease.
Olfactory dysfunction is an early symptom of neurodegenerative disease. Amyloid-beta oligomers (AβOs), the pathologic protein of Alzheimer's disease (AD), have been confirmed to be firstly deposited in olfactory bulb (OB), causing smell to malfunction. However, the detailed mechanisms underlying pathogenic nature of AβOs-induced olfactory neuronal degeneration in AD are not completely realized. Here, an early-stage olfactory dysfunction pathological model of AD in vitro based on biomimetic OB neuronal network chip was established for dynamic multi-site detection of neuronal electrical activity and network connection. We found both spike firing and correlation of overall neuronal network change regularly displayed gradually active state and then rapidly decay state after AβOs induction. Moreover, MK-801 and memantine were administrated at early-stage to detect alteration of OB neurons simulating nasal administration for AD treatment, which showed an almost recovery through the intermittent firing pattern. Together, this neuronal network-on-chip has revealed synaptic impairment and network neurodegeneration of olfactory dysfunction in AD, providing potential mechanisms information for early-stage progressive olfactory amyloidogenic pathology.


Live-interactive teledermatology program in Taiwan: One-year experience serving a district hospital in rural Taitung County.
Population aging and unequal accessibility of health care are increasingly important in developed countries. One strategy to overcome these issues is utilizing telemedicine, which is recently made possible technologically by the advancement of internet speed, high speed zooming cameras, and the information storages. In Taiwan, the telemedicine is granted legally by the amendment for Taiwan's Physician Act in 2018. Kaohsiung Chang Gung Memorial Hospital (Kaohsiung CGMH) is the first hospital in Taiwan to provide the telemedicine service connecting to Cheng Kung Branch of Taitung Hospital since Nov 2018. Consultation services from Dermatology, ENT, and Ophthalmology have been delivered in the live-interactive and face to face module every week. Dermatology consultation comprises the majorities. In the first year, there were totally 426 dermatology consultation services. Eczema, fungal infections, and scabies infestation were the three most common diseases in the beginning. The disease diagnosis became more diverse after several months, including some ready-to-treat diseases pending correct diagnosis, such as pediculosis, psoriasis, and urticaria. Coupled with dermoscopic images, diseases such as hair loss, pediculosis capitis, skin tumor, and scabies, were diagnosed promptly. The subjective patient improvement rate was more than 75% year-round and the case closure rate was more than 85% year-round. Teledermatology is a promising approach to serve the remote medical-underprivileged regions. The teledermatology is anticipated to help underserved regions, nursing homes, prisons, and in situations with severe pandemic infections, such as COVID-19.


SS18-SSX-Dependent YAP/TAZ Signaling in Synovial Sarcoma.
Synovial sarcoma is a soft tissue malignancy characterized by a reciprocal t(X;18) translocation. The chimeric SS18-SSX fusion protein acts as a transcriptional dysregulator representing the major driver of the disease; however, the signaling pathways activated by SS18-SSX remain to be elucidated to define innovative therapeutic strategies. Immunohistochemical evaluation of the Hippo signaling pathway effectors YAP/TAZ was performed in a large cohort of synovial sarcoma tissue specimens. SS18-SSX dependency and biological function of the YAP/TAZ Hippo signaling cascade were analyzed in five synovial sarcoma cell lines and a mesenchymal stem cell model in vitro. YAP/TAZ-TEAD-mediated transcriptional activity was modulated by RNAi-mediated knockdown and the small-molecule inhibitor verteporfin. The effects of verteporfin were finally tested in vivo in synovial sarcoma cell line-based avian chorioallantoic membrane and murine xenograft models as well as a patient-derived xenograft. A significant subset of synovial sarcoma showed nuclear positivity for YAP/TAZ and their transcriptional targets FOXM1 and PLK1. In synovial sarcoma cells, RNAi-mediated knockdown of SS18-SSX led to significant reduction of YAP/TAZ-TEAD transcriptional activity. Conversely, SS18-SSX overexpression in SCP-1 cells induced aberrant YAP/TAZ-dependent signals, mechanistically mediated by an IGF-II/IGF-IR signaling loop leading to dysregulation of the Hippo effectors LATS1 and MOB1. Modulation of YAP/TAZ-TEAD-mediated transcriptional activity by RNAi or verteporfin treatment resulted in significant growth inhibitory effects in vitro and in vivo. Our preclinical study identifies an elementary role of SS18-SSX-driven YAP/TAZ signals, highlights the complex network of oncogenic signaling pathways in synovial sarcoma pathogenesis, and provides evidence for innovative therapeutic approaches.


Transperineal versus transvaginal ultrasound cervical length measurement and preterm labor.
The aim was to evaluate the agreement between and the reproducibility of transperineal and transvaginal ultrasound cervical length measurements performed by the duty obstetrical team in case of preterm labor. The acceptability of transperineal ultrasonography was also assessed. Pregnant patients between 25 and 34 weeks of gestation with contractions and a clinically modified cervix were included. Order of ultrasonography examination (transperineal or transvaginal first) and rank of operator (resident or senior) were allocated randomly. Agreement was assessed using the intraclass correlation coefficient (ICC) and the Bland and Altman plot. The patient's discomfort and preference for either method were assessed with a questionnaire. 62 patients admitted for preterm labor between 25 and 34 weeks of gestation were included. Six seniors and nine residents took part in the study. Among the 51 patients with an interpretable transperineal ultrasound scan, median cervical length measurements with the transperineal and the transvaginal technique were, respectively, 25 mm (0-53) and 27 mm (4-51). Concordance was good with an ICC of 0.83 [IC 95 % = (0.73-0.90)]. Transperineal ultrasonography was preferred in 56.5 % of cases. In case of preterm labor, cervical length measurement with transperineal ultrasonography seems reproducible and can be performed by the obstetric team on duty.


[Microsurgical soft tissue transplantation].
Since the first microsurgical transplantation in 1971, numerous tissue regions which are appropriate for transplantation have been researched. In particular, cutaneous or fasciocutaneous and musculocutaneous, as well as jejunum and fat tissue transplants are involved. In maxillofacial surgery, predominantly radial underarm flaps and scapular or parascapular flaps are used. The indications for both kinds of flaps range from intraoral defects, including tongue defects, and defects after maxillary resection, to defects following tumor resection of the face. In the framework of radical tumor surgery involving regional lymph drainage ducts, the musculus latissimus dorsi flaps and the musculus rectus abdominis flaps are indicated for the covering of deep-reaching defects. Jejunum transplants are appropriate mainly in the region of the soft palate and pharyngeal walls. Free microsurgical fat tissue transplants are indicated for extensive subcutaneous defects. The technique of lifting transplants and transferring them is so refined that results are comparable to those attained by using conventional flaps.


Is radiologists' volume of mammography reading related to accuracy? A critical review of the literature.
The current UK quality assurance guidelines for radiologists in the NHS breast screening programme require those reporting screening mammograms to read a minimum of 5000 cases per year. We aimed to review the evidence for this and to assess whether there was justification for lowering the required level. A literature search was conducted to identify relevant studies where accuracy of reporting mammograms was related to reading volume. Three of the five studies reviewed suggested a positive association between reading volume and sensitivity, but there were few data on volumes above 5000 cases per year. The available evidence did not provide any basis for reducing the threshold volume. Further work is needed, in a UK or European setting, to study the relationship between reading volume and accuracy at higher volume levels and also the separate effects of reading volume and reading experience.


Human T Cells Expressing a CD19 CAR-T Receptor Provide Insights into Mechanisms of Human CD19-Positive β Cell Destruction.
Autoimmune destruction of pancreatic β cells underlies type 1 diabetes (T1D). To understand T cell-mediated immune effects on human pancreatic β cells, we combine β cell-specific expression of a model antigen, CD19, and anti-CD19 chimeric antigen receptor T (CAR-T) cells. Coculturing CD19-expressing β-like cells and CD19 CAR-T cells results in T cell-mediated β-like cell death with release of activated T cell cytokines. Transcriptome analysis of β-like cells and human islets treated with conditioned medium of the immune reaction identifies upregulation of immune reaction genes and the pyroptosis mediator GSDMD as well as its activator CASP4. Caspase-4-mediated cleaved GSDMD is detected in β-like cells under inflammation and endoplasmic reticulum (ER) stress conditions. Among immune-regulatory genes, PDL1 is one of the most upregulated, and PDL1 overexpression partially protects human β-like cells transplanted into mice. This experimental platform identifies potential mechanisms of β cell destruction and may allow testing of therapeutic strategies.


Shoulder adduction contracture after burn: anatomy and treatment with quadrangular local scar subcutaneous pedicled flap, a new approach.
Axillary adduction contracture is caused by scars that tightly surround the shoulder joint impairing the function of the upper limb. Due to severe scar surface deficiency, contracture release presents a challenge for surgeons since a method of release is transfer of tissue in the form of a large pedicled or free flap(s). Thus, development of simpler, less traumatic techniques, using local tissues, persists. Anatomic studies of shoulder adduction contractures after burn (pre-operative, during surgery, post-reconstruction) were done in 346 pediatric and adult patients. All were divided into three groups according to contracture types: with edge contractures (80%), medial (6%) and total (14%). Anatomical study covered peculiarities of total contractures and possibilities for their treatment using local scarred tissue. Total contractures (48 patients) were caused by scars tightly surrounding the joint on three sides: anterior, posterior, and axillary. There were two specific forms of contracture: (a) shoulder close to the chest wall (22 of 48 patients) which was treated with thoracic pedicled or free flaps; (b) in 26 out of 48 patients a flat scar and skin graft surface laid along the shoulder and chest wall, in axillary projection, which were used for contracture release in the form of a subcutaneous pedicled quadrangular flap. The flap was mobilized only peripherally, descending to the apex of the axilla, forming the central axillary zone, and suspension of the axilla on a normal level. Wounds aside the flaps were covered with skin graft. Acceptable functional and cosmetic results were achieved in all 26 patients. Total shoulder adduction contractures have two forms: (a) shoulder close/fused with the chest wall; and (b) along the chest wall and shoulder there is a flat surface, the tissue of which can be used for reconstruction in a form of scar subcutaneous pedicled quadrangular flap. Based on this flap, a new technique is described which is relatively easy to perform.


Clinical characteristics and short-term outcomes of Japanese encephalitis in pediatric and adult patients: a retrospective study in Northern China.
The study aimed to compare the clinical characteristics and short-term outcomes of pediatric and adult Japanese encephalitis (JE) patients in order to find out the differences. From August 2006 to October 2019, 107 patients (62 pediatric patients and 45 adult patients) with JE were enrolled. Clinical characteristics and short-term outcomes were analyzed. The short-term outcome of each patient was defined as a good outcome or poor outcome according to their Glasgow Coma Scale (GCS) scores (GCS > 8 vs. GCS ≤ 8) at discharge. As for acute complications, the incidence of pulmonary infection was higher in 25 adults (25/45, 55.6%) than in 19 children (19/62, 30.6%; P = 0.01). Upper gastrointestinal bleeding was more common in patients with pulmonary infection, with 10 of these patients experiencing the symptom (10/44, 22.7%) compared to only one patient without pulmonary infection (1/63, 1.6%; P = 0.001). The proportion of mechanical ventilation and admission to the intensive care unit (ICU) for supportive care was higher in patients with pulmonary infection than in patients without infection (P < 0.001, P = 0.008, respectively). The GCS scores at discharge in patients with pulmonary infection (7, 4-12.75) were lower than in patients without pulmonary infection (14, 10-14; P < 0.001). Although the GCS scores at the admission of children (9.5, 7-13) were similar to that of adults (7, 6-13), the GCS scores at the discharge of adults (7, 3.5-13) were lower than that of children (13, 10.75-14; P < 0.001). The short-term outcome of JE was worse in adults. Pulmonary infection was correlated with a high incidence of upper gastrointestinal bleeding, mechanical ventilation, and ICU hospitalization in JE. Pulmonary infection is a prognostic predictor of short-term outcomes in patients with JE. Vaccination for adults should be initiated.


Risk of cranial ultrasound abnormalities in very-low-birth-weight infants conceived with assisted reproductive techniques.
To evaluate risks of cranial ultrasound abnormalities among very-low-birth-weight (VLBW) infants conceived with fertility therapy (ovulation induction only or with assisted reproductive techniques [ART]) and of multiple gestation pregnancies. The incidences of cranial ultrasound abnormalities in 1473 VLBW infants conceived with and without fertility therapy and born of multiple versus singleton pregnancies were compared, using logistic regression models. Infants conceived with ART were less likely to have intraventricular hemorrhage (IVH). Twins and triplets had risks of cranial ultrasound abnormalities similar to those of singletons. Twins and triplets conceived with ART were at lower risk of IVH. VLBW infants conceived with ART do not appear to be at increased risk of cranial ultrasound abnormalities. Likewise, twins and triplets were not at increased risk of these abnormalities.


Aromatic dioxygenases: molecular biocatalysis and applications.
Aromatic dioxygenases have been found to catalyse single and tandem oxidation reactions of conjugated polyenes. Rational selection and design of dioxygenases, allied to substrate shape, size and substitution pattern, has been used to control regiochemistry and stereochemistry during the oxygenation process. The resulting enantiopure bioproducts have been increasingly utilised as precursors for new and alternative routes in chiral synthesis.


[Effect of YXXM motif on viral replication of subgroup J avian leukosis virus].
It was reported that subgroup J avian leukosis virus strain NX0101 activates PI3K/Akt pathway during early infection in DF-1 cells. Whether there is YXXM motif in the amino acid sequence of NX0101 and the function of YXXM motif were studied. The presence of internal transmembrane domains in the envelope protein of NX0101 was analyzed by Tmpred. Point mutation was introduced to change the YXXM motif in the NX0101 strain to FXXA. The plasmid containing the full genome of NX0101 with mutation within the YXXM motif in pMD18-T vector was constructed and transfected into DF-1 cells. Viral replication levels of NX0101 strain and the mutation one were tested and compared by real-time PCR and ELISA. The amino acid sequence of NX0101 strain had one YXXM motif (amino acids 554-557) in the cytoplasmic tail of envelope protein. The mutated NX0101 strain (Y554F, M557A) was rescued by reverse genetics technique. Viral replication of the mutated NX0101 strain was significantly lower than that of NX0101 strain in the level of either RNA or protein synthesis. The results revealed that the (Y)XXM motif was important for virus NX0101 replication in DF-1 cells.


Insect Migration and Changes in Venom Allergy due to Climate Change.
Insects are highly successful animals. They have limited ability to regulate their temperature and therefore will expand range in response to warming temperatures. Climate change and associated rising global temperature is impacting the range and distribution of stinging insects. There is evidence that many species are expanding range toward the poles, primarily in response to warming. With expanded distribution of stinging insects, increased interaction with humans is anticipated with consequently increased rates of sting-related reactions and need for intervention. This article focuses on evidence that insects are expanding their range in response to warming temperature, increasing likelihood of human interaction.


Structural basis of the TAM domain of BAZ2A in binding to DNA or RNA independent of methylation status.
Bromodomain adjacent to zinc finger domain protein 2A (BAZ2A) (also called transcription termination factor-1 interacting protein 5), a key component of the nucleolar remodeling complex, recruits the nucleolar remodeling complex to ribosomal RNA genes, leading to their transcriptional repression. In addition to its tandem plant homeodomain-bromodomain that is involved in binding to acetylated histone H4, BAZ2A also contains a methyl-CpG-binding domain (MBD)-like Tip5/ARBP/MBD (TAM) domain that shares sequence homology with the MBD. In contrast with the methyl-CpG-binding ability of the canonical MBD, the BAZ2A TAM domain has been shown to bind to promoter-associated RNAs of ribosomal RNA genes and promoter DNAs of other genes independent of DNA methylation. Nevertheless, how the TAM domain binds to RNA/DNA mechanistically remains elusive. Here, we characterized the DNA-/RNA-binding basis of the BAZ2A TAM domain by EMSAs, isothermal titration calorimetry binding assays, mutagenesis analysis, and X-ray crystallography. Our results showed that the TAM domain of BAZ2A selectively binds to dsDNA and dsRNA and that it binds to the backbone of dsDNA in a sequence nonspecific manner, which is distinct from the base-specific binding of the canonical MBD. Thus, our results explain why the TAM domain of BAZ2A does not specifically bind to mCG or TG dsDNA like the canonical MBD and also provide insights for further biological study of BAZ2A acting as a transcription factor in the future.


[A man with a yellow coloured abnormality of his finger].
A 83-year-old man presented to the emergency department with signs of diabetes mellitus de novo and a yellow coloured abnormality of his left index finger. A plain radiograph of the left hand showed destruction of the proximal interphalangeal joint of digit II. With use of polarised light microscopy, the diagnosis 'gouty tophus' was established.


Chronic Spontaneous Urticaria: An Etiopathogenic Study.
Exact etiopathogenesis of chronic spontaneous urticaria (CSU) remains elusive. Infections, pseudoallergens, autoimmunity, and contact sensitization are various postulated factors. Few studies are available measuring cytokine levels in CSU. The aim was to study various etiological factors of CSU and levels of IL-6 and IFN-ϒ in cases and controls, and correlation between various etiologies with the levels of the abovementioned interleukins in the cases. Case-control study performed over 2 years with no follow-up of the participants. It was a referral-center-based study. Sixty patients of CSU and equal age and sex-matched healthy controls were recruited on the basis of convenience sampling. Biochemical and hematological investigations with hepatitis serology, thyroid function tests, anti-thyroid antibodies, and levels IL-6 and IFN-ϒ were performed in all cases and controls. All cases were subjected to ASST. Cases with all above negative tests were patch-tested with Indian standard series. Urticaria activity score (UAS7) was calculated for all the cases and repeated in patients with positive etiological factor after 3 weeks (improvement after allergen or drug avoidance, treatment of infection). To study the various etiological factors (food, infection, autoimmunity, autoreactivity, and contact sensitization) and the levels of IL-6 and IFN- γ in patients of chronic spontaneous urticaria. Etiology was ascertained in 75% of patients (autoimmunity: 50%, contact sensitization: 21.67%, food and drug allergy: 1.67% each). Mean values of the interleukins and anti-thyroid antibodies were significantly higher in cases versus controls. Levels of IFN-ϒ were significantly elevated in patients with higher UAS7 scores. Antithyroid antibodies, ASST, and patch testing are important tools and should be considered in patients of CSU after a thorough history and history-based workup. Elevated levels of IL-6 and IFN- ϒ in cases suggest that both Th1 and Th2 type of immune responses are implicated in pathogenesis of CSU.


The GABA(B) antagonist CGP 52432 attenuates the stimulatory effect of the GABA(B) agonist SKF 97541 on luteinizing hormone secretion in the male sheep.
The objectives of this study were to determine if the gamma-aminobutyric acid (GABA)(B) agonist, 3-aminopropyl (methyl) phosphinic acid (SKF97541), would increase luteinizing hormone (LH) secretion when infused by microdialysis into the medial basal hypothalamus (MBH) of the castrated ram, and to determine if the action of SKF97541 would be attenuated by co-infusion of the GABA(B) antagonist CGP52432. Initial experiments established that infusion of SKF alone, at concentrations as low as 5 microM, increased mean LH, LH pulse amplitude, and in some cases, pulse interval. In the last experiment, animals were treated with artificial cerebrospinal fluid (CSF) alone, SKF alone (30 microM), 3-[[(3, 4-dichlorophenol) methyl] amino] propyl] diethoxymethyl) phosphinic acid (CGP) alone (500 microM), or SKF plus CGP. SKF increased both mean LH and LH pulse amplitude as compared with CSF. CGP alone had no significant effect on LH, but it attenuated the effect of SKF on mean LH. These observations indicate that the stimulatory effects of GABA(B) agonists on LH pulse patterns are mediated through GABA(B) receptors and provide further evidence that GABA(B) receptors located in the MBH can regulate pulsatile GnRH-LH release.


Eruptive angiokeratomas and porokeratosis in the setting of sclerodermatous graft-vs.-host disease.
Porokeratosis and angiokeratomas are both seen in the setting of chronic graft-vs.-host disease (GVHD), but rarely occur together. We present a case of a patient with lichen planus-like (LPL) and sclerodermatous chronic GVHD manifesting after allogeneic bone marrow transplant with concomitant eruptive angiokeratomas and porokeratosis.


Extraction of DNA from soil using nanoparticles by magnetic bioseparation.
To develop a simple, rapid and inexpensive soil DNA extraction protocol. The protocol relies on the use of superparamagnetic silica-magnetite nanoparticles for the isolation and purification of DNA from soil samples. DNA suitable for use in molecular biology applications was obtained from a number of soil samples. The DNA extracted using the tested method successfully permitted the PCR amplification of a fragment of the bacterial 16S rDNA gene. The extracted DNA could also be restriction endonuclease digested. The protocol reported here is simple and permits rapid isolation of PCR-ready soil DNA. The method requires only small quantities of soil sample, is scalable and suitable for automation.


Recognition of whole Cryptosporidium oocysts in feces by negative staining and electron microscopy.
Crytosporidium oocysts in feces were recognizable in the electron microscope when prepared by the negative staining technique used by virologists. Their size, shape, and surface markings were sufficiently characteristic for cryptosporidiosis to be diagnosed by this method if more suitable methods are not available.


Fate and toxicity of melamine in activated sludge treatment systems after a long-term sludge adaptation.
Melamine is a nitrogen-rich (67% nitrogen by mass) heterocyclic aromatic compound that could significantly increase effluent total nitrogen concentrations. In this study, we investigated the degradation of melamine and its impact on activated sludge operations by employing two common activated sludge processes, namely the Modified Ludzack-Ettinger (MLE) process and the continuous stirred tank reactor (CSTR) process. Melamine was dosed continuously from day 125 in both activated sludge treatment systems at an influent concentration of 3 mg/L for about 100 days. Even after such a long period of sludge adaptation, melamine appeared not to be easily biodegradable. The average melamine removal efficiencies in the CSTR and MLE systems were 14 ± 10% and 20 ± 15%, respectively. There was no significant difference in melamine removal between the two different activated sludge processes. The long-term input of melamine resulted in a decrease in the nitrifying bacterial activities (by 82 ± 8%) and population in both systems. Short-term microtiter assay results also showed that melamine reduced activated sludge growth by 80% when supplied at a concentration of 75.6 mg/L. These results suggest that sludge adaptation plays a minimal role in melamine degradation, as the enzymes responsible for hydrolytic deamination of melamine in activated sludge are not easily induced. The insignificant biodegradation of melamine is also attributed to bacterial growth inhibition under long-term dosing conditions with melamine, resulting in a significant decrease in effluent water quality.


Key roles of β-glucosidase BglA for the catabolism of both laminaribiose and cellobiose in the lignocellulolytic bacterium Clostridium thermocellum.
The thermophilic bacterium Clostridium thermocellum efficiently degrades polysaccharides into oligosaccharides. The metabolism of β-1,4-linked cello-oligosaccharides is initiated by three enzymes, i.e., the cellodextrin phosphorylase (Cdp), the cellobiose phosphorylase (Cbp), and the β-glucosidase A (BglA), in C. thermocellum. In comparison, how the oligosaccharides containing other kinds of linkage are utilized is rarely understood. In this study, we found that BglA could hydrolyze the β-1,3-disaccharide laminaribiose with much higher activity than that against the β-1,4-disaccharide cellobiose. The structural basis of the substrate specificity was analyzed by crystal structure determination and molecular docking. Genetic deletions of BglA and Cbp, respectively, and enzymatic analysis of cell extracts demonstrated that BglA is the key enzyme responsible for laminaribiose metabolism. Furthermore, the deletion of BglA can suppress the expression of Cbp and the deletion of Cbp can up-regulate the expression of BglA, indicating that BglA and Cbp have cross-regulation and BglA is also critical for cellobiose metabolism. These insights pave the way for both a fundamental understanding of metabolism and regulation in C. thermocellum and emphasize the importance of the degradation and utilization of polysaccharides containing β-1,3-linked glycosidic bonds in lignocellulose biorefinery.


Comparing prevalence estimates of concussion/head injury in U.S. children and adolescents in national surveys.
Reports on pediatric lifetime concussions/head injuries (LCHI) from national surveys have offered estimates on prevalence that range from 2.5% to 18% in the general population. The purpose of this study is to examine national surveys to compare methodologies and limitations pertaining to LCHI data collection. Three nationally representative surveys that measure LCHI in children, including the National Survey of Children's Health, the National Health Interview Survey, and the Monitoring the Future Survey were examined. Children were grouped by ages 3-17 years and adolescent ages 13-17 years, stratified by selected demographic characteristics. Participants in the surveys included parents (NSCH and NHIS) and adolescents (MTF survey). The primary outcome measure is an estimate of LCHI in children. Estimates of prevalence of LCHI ranged from 3.6% to 7.0% for children ages 3-17 years and from 6.5% to 18.3% for adolescents 13-17 years. Survey modality, question wording, and respondent may contribute to differing estimates. Prevalence showed consistent variation by age, sex, and race/ethnicity across surveys. Associations were inconsistent between LCHI and insurance status, parental education, and household primary language. Although there are methodological differences in capturing pediatric LCHI across surveys, the prevalence estimates and correlational associations generated can offer awareness about the burden of these injuries and insights to research and clinical care.