Drug-induced aseptic meningitis: a mini-review.
Aseptic meningitis associates a typical clinical picture of meningitis with the absence of bacterial or fungal material in the cerebrospinal fluid. Drug-induced aseptic meningitis (DIAM) may be due to two mechanisms: (i) a direct meningeal irritation caused by the intrathecal administration of drugs and (ii) an immunologic hypersensitivity reaction to a systemic administration. If the direct meningeal irritation allows a rather easy recognition, the immunologic hypersensitivity reaction is a source of challenging diagnostics. DIAM linked to a systemic treatment exerts typically an early onset, usually within a week. This period can be shortened to a few hours in case of drug rechallenge. The fast and spontaneous regression of clinical symptoms is usual after stopping the suspected drug. Apart from these chronological aspects, no specific clinical or biological parameters are pathognomonic. CSF analysis usually shows pleiocytosis. The proteinorachia is increased while glycorachia remains normal. Underlying pathologies can stimulate the occurrence of DIAM. Thus, systemic lupus erythematosus appears to promote DIAM during NSAID therapy, especially ibuprofen-based one. Similarly, some patients with chronic migraine are prone to intravenous immunoglobulin-induced aseptic meningitis. DIAM will be mainly evoked on chronological criteria such as rapid occurrence after initiation, rapid regression after discontinuation, and recurrence after rechallenge of the suspected drug. When occurring, positive rechallenge may be very useful in the absence of initial diagnosis. Finally, DIAM remains a diagnosis of elimination. It should be suggested only after all infectious causes have been ruled out.


Host preferences inhibit transmission from potential superspreader host species.
Host species that are particularly abundant, infectious and/or infected tend to contribute disproportionately to symbiont (parasite or mutualist) maintenance in multi-host systems. Therefore, in a facultative multi-host system where two host species had high densities, high symbiont infestation intensities and high infestation prevalence, we expected interspecific transmission rates to be high. Instead, we found that interspecific symbiont transmission rates to caged sentinel hosts were an order of magnitude lower than intraspecific transmission rates in the wild. Using laboratory experiments to decompose transmission rates, we found that opportunities for interspecific transmission were frequent, where interspecific and intraspecific contact rate functions were statistically indistinguishable. However, most interspecific contacts did not lead to transmission events owing to a previously unrecognized transmission barrier: strong host preferences. During laboratory choice experiments, the symbiont preferred staying on or dispersing to its current host species, even though the oligochaete symbiont is a globally distributed host generalist that can survive and reproduce on many snail host species. These surprising results suggest that when managing symbiont transmission, identifying key host species is still important, but it may be equally important to identify and manage transmission barriers that keep potential superspreader host species in check.


High-Soluble-Fiber Diet Attenuates Hypoxia-Induced Vascular Remodeling and the Development of Hypoxic Pulmonary Hypertension.
Hypoxic pulmonary hypertension is a difficult disease to manage that is characterized by sustained elevation of pulmonary vascular resistance and pulmonary artery pressure due to vasoconstriction, perivascular inflammation, and vascular remodeling. Consumption of soluble-fiber is associated with lower systemic blood pressure, but little is known about its ability to affect the pulmonary circulation. Mice were fed either a low- or high-soluble-fiber diet (0% or 16.9% inulin) and then exposed to hypoxia (FiO2, 0.10) for 21 days to induce pulmonary hypertension. The impact of diet on right ventricular systolic pressure and pulmonary vascular resistance was determined in vivo or in ex vivo isolated lungs, respectively, and correlated with alterations in the composition of the gut microbiome, plasma metabolome, pulmonary inflammatory cell phenotype, and lung proteome. High-soluble-fiber diet increased the abundance of short-chain fatty acid-producing bacteria, with parallel increases in plasma propionate levels, and reduced the abundance of disease-related bacterial genera such as Staphylococcus, Clostridioides, and Streptococcus in hypoxic mice with parallel decreases in plasma levels of p-cresol sulfate. High-soluble-fiber diet decreased hypoxia-induced elevations of right ventricular systolic pressure and pulmonary vascular resistance. These changes were associated with reduced proportions of interstitial macrophages, dendritic cells, and nonclassical monocytes. Whole-lung proteomics revealed proteins and molecular pathways that may explain the effect of soluble-fiber supplementation. This study demonstrates for the first time that a high-soluble-fiber diet attenuates hypoxia-induced pulmonary vascular remodeling and the development of pulmonary hypertension in a mouse model of hypoxic pulmonary hypertension and highlights diet-derived metabolites that may have an immuno-modulatory role in the lung.


Prediction of protein flexibility using a conformationally restrained contact map.
Knowledge of protein flexibility is crucial to understanding protein function. However, probing protein flexibility by either experiment or computational simulations is a difficult process. In particular, many computational approaches to understanding protein flexibility require an experimentally determined protein structure. The Conformationally Restrained Contact Map (CoRe-CMap) approach reported here couples protein disorder predictions with protein structure predictions and only requires sequence data to predict protein flexibility. This paper reports the application of the CoRe-CMap model to predicting Lipari-Szabo order parameters of all proteins for which experimentally derived Lipari-Szabo order parameters are available in the BioMagResBank: the median root mean square deviation between a protein's predicted and experimentally derived order parameters is 0.124. Additionally, application of the CoRe-CMap model to predict Lipari-Szabo order parameters for the 10th Type III Domain in Fibronectin and a homologous domain from Tenascin demonstrates the ability of CoRe-CMap to predict functionally important differences in protein flexibility.


Transgender Scrotoplasty and Perineal Reconstruction With Labia Majora Flaps: Technique and Outcomes From 147 Consecutive Cases.
Labia majora, the embryologic homologs of the scrotum, are ideal donor tissue for transgender scrotoplasty. The technique is detailed, and surgical outcomes are assessed for scrotoplasty using labia majora rotational advancement flaps. We retrospectively reviewed the outcomes of phalloplasty patients who underwent either primary or secondary labia majora flap scrotoplasty and perineal reconstruction from October 1, 2017, to December 1, 2019. Bilateral elevation and rotational flap advancement from the posterior to anterior position formed a pouch-like scrotum. Perineal reconstruction involved multilayered closure with apposition of the inner thigh skin. The mean follow-up was 12.5 months (0.5-26 months). One hundred forty-seven scrotoplasties were performed. Of the 147 total scrotoplasty patients, 133 had labia majora flap scrotoplasty and perineal reconstruction with single-stage phalloplasty. Distal flap necrosis occurred in 6 patients (4.1%); 5 were ipsilateral to the groin dissection required for phalloplasty. Large (>1 cm diameter) perineoscrotal junction dehiscence occurred in 7 patients (4.7%). All wounds were managed conservatively except for 3 patients who developed urethrocutaneous fistulas at the perineoscrotal junction. All 3 patients required fistula repair. Two (1.4%) scrotal hematomas and 3 (2.0%) perineal hematomas were seen; all required operative intervention. Labia majora flap scrotoplasty via the bilateral rotational advancement technique and perineal reconstruction can be safely performed during phalloplasty. Minor wound complications are common and frequently heal with conservative management. Wounds that do not heal may be associated with urethral complications. Hematomas are rare but usually require operative intervention.


Control of expression of major histocompatibility complex genes in horse trophoblast.
In most mammals, the fetus limits its presentation of paternal antigens to the mother by suppressing the cell-surface expression of proteins of the major histocompatibility complex (MHC) on trophoblast. In the horse, however, functional, polymorphic MHC class I antigens are expressed at high levels on the invasive trophoblast cells of the chorionic girdle between Days 32 and 36 of pregnancy, although not on the adjacent noninvasive trophoblast of the chorion and allantochorion membranes. In this study, the control of MHC class I gene expression was investigated in invasive and noninvasive horse trophoblast, and the MHC class I loci expressed by invasive trophoblast were identified. Northern blot hybridization of Day 33-34 conceptus tissue revealed both transcriptional and posttranscriptional regulation of cell-surface MHC class I expression in horse trophoblast. The invasive MHC class I-positive trophoblast showed levels of steady-state mRNA nearly as high as those in lymphoid tissues from adult horses, whereas noninvasive MHC class I-negative trophoblast also contained transcripts for MHC class I, but at lower levels similar to those present in adult horse nonlymphoid tissue. We also cloned and sequenced polymerase chain reaction products from the transmembrane and cytoplasmic regions of MHC class I transcripts in chorionic girdle and lymphocytes, and determined that horse invasive trophoblast appears to transcribe the same MHC class I loci transcribed in lymphocytes, including both polymorphic and nonpolymorphic loci. These data from the horse demonstrate that functional alloantigen presentation by trophoblast can be a normal part of early pregnancy.


Effect of Statins, Metformin, Angiotensin-Converting Enzyme Inhibitors, and Angiotensin II Receptor Blockers on Age-Related Macular Degeneration.
Statins, metformin, angiotensin-converting enzyme (ACE) inhibitors, and angiotensin receptor blockers (ARBs) have been suggested for treating age-related macular degeneration (AMD) due to their pleiotropic effects. Therefore, we investigated whether these drugs prevent AMD. We conducted a nested case-control study using the Korean National Health Insurance Service database. Using risk-set sampling of age, sex, cohort entry date, and follow-up duration, we identified incident patients with AMD and 10 matching controls in cohorts with diabetes mellitus or cardiovascular diseases. Exposure was assessed within one year before the index date using patient prescription records. We conducted conditional logistic regression to estimate the odds ratios (ORs) and 95% confidence intervals (CIs) to evaluate the association between cardiovascular medications and AMD. Our study included 2330 cases and 23278 controls from a cohort of 231274 patients. The ORs (95% CI) for AMD occurrence in users prescribed with statins, metformin, ACE inhibitors, and ARBs were 1.12 (0.94-1.32), 1.15 (0.91-1.45), 0.90 (0.61-1.34), and 1.21 (1.05-1.39), respectively. A duration-response was not observed. Statins, metformin, ACE inhibitors, and ARBs did not inhibit AMD in elderly patients. The absence of a duration-response supports the lack of a causal relationship.


New concepts of allergy to cow's milk.
CMA is a diagnostic and therapeutic problem in the field of pediatrics. The response to oral cow's milk challenge was evaluated in patients with suspective CMA. To investigate immune mechanisms in different subtypes of CMA, suppressor activity, in vitro IFN-gamma and IL-4 generation, eosinophil activation and humoral immune response were measured. Also, changes in the function of the immune system were evaluated when patients had clinically recovered from CMA. The results of different measurements were correlated with the patients' clinical response to cow's milk challenge. The prechallenge suppressor activity was found to be low in patients with challenge-proven CMA when compared with those clinically negative to oral cow's milk challenge but had normalized after a 4-18 month milk-elimination period in patients who had recovered from CMA. IFN-gamma generation was low in patients with active CMA but had clearly enhanced in patients recovering from CMA. The serum concentration of ECP increased significantly during oral cow's milk challenge in patients with cutaneously manifested CMA. Measured by the ELISPOT method, the patients with active CMA mounted a high, non-antigen-specific immune response but were unable to direct the antigen-specific response, especially in the IgA class. After a follow-up of a mean 13.5 months on cow's milk elimination diet, the immune response to cow's milk antigens had developed in patients who had recovered from CMA. The development in the capacity to mount an adequate immune response to specific protein antigens, especially in the IgA class, can be interpreted as deriving from the development in T-lymphocyte regulation.(ABSTRACT TRUNCATED AT 250 WORDS)


Virtual crystal description of III-V semiconductor alloys in the tight binding approach.
We propose a simple and effective approach to construct the empirical tight-binding parameters of ternary alloys in the virtual crystal approximation. This combines a new, compact formulation of the strain parameters and a linear interpolation of the Hamiltonians of binary materials strained to the alloy equilibrium lattice parameter. We show that it is possible to obtain a perfect description of the bandgap bowing of ternary alloys in the InGaAsSb family of materials. Furthermore, this approach is in a good agreement with supercell calculations using the same set of parameters. This scheme opens a way for atomistic modeling of alloy-based quantum wells and quantum wires without extensive supercell calculations.


Molecular transitions in early progenitors during human cord blood hematopoiesis.
Hematopoietic stem cells (HSCs) give rise to diverse cell types in the blood system, yet our molecular understanding of the early trajectories that generate this enormous diversity in humans remains incomplete. Here, we leverage Drop-seq, a massively parallel single-cell RNA sequencing (scRNA-seq) approach, to individually profile 20,000 progenitor cells from human cord blood, without prior enrichment or depletion for individual lineages based on surface markers. Our data reveal a transcriptional compendium of progenitor states in human cord blood, representing four committed lineages downstream from HSC, alongside the transcriptional dynamics underlying fate commitment. We identify intermediate stages that simultaneously co-express "primed" programs for multiple downstream lineages, and also observe striking heterogeneity in the early molecular transitions between myeloid subsets. Integrating our data with a recently published scRNA-seq dataset from human bone marrow, we illustrate the molecular similarity between these two commonly used systems and further explore the chromatin dynamics of "primed" transcriptional programs based on ATAC-seq. Finally, we demonstrate that Drop-seq data can be utilized to identify new heterogeneous surface markers of cell state that correlate with functional output.


Targeting of IL-2 to cytotoxic lymphocytes as an improved method of cytokine-driven immunotherapy.
The use of high-dose interleukin-2 (IL-2) has fallen out of favor due to severe life-threatening side effects. We have recently described a unique way of directly targeting IL-2 to cytotoxic lymphocytes using a virally encoded immune evasion protein and an IL-2 mutant that avoids off-target side effects such as activation of regulatory T cells and vascular endothelium.


HEALTH WATCH: health promotion and disease prevention in primary care.
HEALTH WATCH, a longitudinal prospective study of healthy aging, was designed to characterize a healthy population of 2,200 men and women, ages 20-80 years in 1970. Biochemical, hematological, and physiological tests are performed annually over three weekly visits, combined with a self-administered HEALTH WATCH questionnaire to measure health status and behaviors in seven areas (with over 1,330 variables). In 1988, the HEALTH WATCH study was modified to assess characteristics of an oldest old "productive aging" cohort in Kauai, Hawaii. Nutrition, physical activity, extended family, and spirituality were found to be major health determinants. During 1989 to 1991 a controlled intervention study (ten local primary care physicians and their patients, aged 65-89 years) was completed in the Sun Cities, Arizona. These studies provide evidence that primary care physicians can promote positive health outcomes in patients of any chronological age and baseline health status through active healthy aging interventions.


Top 50 most-cited articles on craniovertebral junction surgery.
Craniovertebral junction is a complex anatomical location posing unique challenges to the surgical management of its pathologies. We aimed to identify the fifty most-cited articles that are dedicated to this field. A keyword search using the Thomson Reuters Web of Knowledge was conducted to identify articles relevant to the field of craniovertebral junction surgery. The articles were reviewed based on title, abstract, and methods, if necessary, and then ranked based on the total number of citations to identify the fifty most-cited articles. Characteristics of the articles were determined and analyzed. The earliest top-cited article was published in 1948. When stratified by decade, 1990s was the most productive with 16 articles. The most-cited article was by Anderson and Dalonzo on a classification of odontoid fractures. By citation rate, the most-cited article was by Herms and Melcher who described Goel's technique of atlantoaxial fixation using C1 lateral mass screws and C2 pedicle screws with rod fixation. Atlantoaxial fixation was the most common topic. The United States, Barrow Neurological Institute, and VH Sonntag were the most represented country, institute, and author, respectively. The significant majority of articles were designed as case series providing level IV evidence. Using citation analysis, we have provided a list of the most-cited articles representing important contributions of various authors from many institutions across the world to the field of craniovertebral junction surgery.


Appendiceal ultrasonography performed by nonradiologists: does it help in the diagnostic process?
We performed a retrospective study to compare the sensitivity, specificity, predictive value, and diagnostic accuracy of appendiceal ultrasonography performed by unsupervised technicians during the nighttime hours with studies performed during the day by supervised technicians. Fifty-nine percent of the 163 sonographic examinations were done during the day, and 41% were performed at night. The sensitivity during the day (61%) was significantly higher than at night (26%), as was the positive predictive value (93% day, 71% night). We conclude that ultrasonography is an operator-dependent study. Its sensitivity is so diminished when not performed by an experienced radiologist or technician that a negative examination is not reliable.


Modification of histone by glyoxal: recognition of glycated histone containing advanced glycation adducts by serum antibodies of type 1 diabetes patients.
Dicarbonyl compounds react more rapidly, than glucose, with arginine and lysine in proteins to form advanced glycation end products (AGEs) and further produce free radicals which cause DNA damage. AGEs are reliable diagnostic biomarkers for most of the age-related diseases. In the present study histone was modified with glyoxal and it was characterized by various spectral techniques. Binding characteristics of the modified histone towards serum antibodies from type 1 diabetes patients was evaluated by solid phase enzyme immunoassay and the results were compared with normal human subjects. Fluorescence and Fourier transformed infrared analysis of the nuclear protein clearly indicated changes in their respective intensities upon modification with glyoxal. Liquid chromatography together with mass spectrometry showed new peaks and m/z values related to AGE adducts of dihydroimidazolidines/hydroimidazolones. This glyoxal modified protein was recognized by serum antibodies of the diabetes patients while it showed negligible binding with that of normal human subjects. Glyoxal modification of histone causes structural turbulence and formation of advanced glycation adducts in histone. These adducts might be the main antigenic epitope of the modified histone, leading to its recognition by circulating type 1 diabetes antibodies.


Standardization and optimization of fluorescence in situ hybridization (FISH) for HER-2 assessment in breast cancer: A single center experience.
Accurate assessment of human epidermal growth factor receptor 2 (HER-2) is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE) tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples) and DAKO Histology FISH Accessory Kit (30 samples). The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing) affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70); the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p < 0.0001). The greatest discordance (82%) between IHC and FISH was observed in IHC 2+ group. A standardized FISH protocol for HER-2 assessment, with high hybridization efficiency, is necessary due to variability in tissue processing and individual tissue characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.


Recombinant, B-domain deleted factor VIII (r-VIII SQ): pharmacokinetics and initial safety aspects in hemophilia A patients.
The pharmacokinetics of a second-generation recombinant B-domain deleted factor VIII (FVIII) preparation (r-VIII SQ) were studied in 36 patients with severe hemophilia A. In contrast to full-length recombinant FVIII, no albumin needs to be added to stabilize the final formulation of this B-domain deleted FVIII preparation. The in vivo recovery and half-life of r-VIII SQ were similar to those of plasma-derived (pd) FVIII (mean half-life of r-VIII SQ, 11.7 h). The volume of distribution and clearance were slightly, but significantly, higher for r-VIII SQ than for pdFVIII (p < 0.05). Peak plasma levels of FVIII were consistently related to the administered dose of r-VIII SQ (r = 0.94, p < 0.0001). The pharmacokinetic profile of r-VIII SQ remained essentially unchanged in a dose range of 25-100 IU/kg body weight and could be reproduced after repeated doses. r-VIII SQ was well tolerated. In conclusion, deletion of the B-domain of FVIII does not influence its in vivo pharmacokinetics.


The interpersonal process model of demand/withdraw behavior.
The demand/withdraw interaction pattern is a destructive cycle of relationship communication behavior that is associated with negative individual and relationship outcomes. Demand/withdraw behavior is thought to be strongly linked to partners' emotional reactions, but current theories are inconsistent with empirical findings. The current study proposes the interpersonal process model of demand/withdraw behavior, which includes linkages between each partners' emotional reactions and the interpersonal behavior of demanding and withdrawing. Data come from problem solving discussions of 55 German couples with observationally coded demand/withdraw behavior and fundamental frequency (f₀) to measure vocally encoded emotional arousal. Actor-partner interdependence models (Kenny, Kashy, & Cook, 2006) were used to examine associations among demand/withdraw behavior and f₀ in the overall discussion and 5-min segments. Significant cross-partner associations emerged for demanding and withdrawing behavior across the whole conversation as well as within 5-min segments, and these associations are partially accounted for by each individual's f₀. When behaviorally coded demanders expressed more vocal arousal, they demanded more and withdrew less while their partners withdrew more. In contrast, when behaviorally coded withdrawers expressed more vocal arousal, their partners demanded less and withdrew more. Findings demonstrate that demand/withdraw behavior varies between couples (i.e., some couples engage in a stronger demand/withdraw cycle than others) and between segments (i.e., when 1 partner increases demanding, the other increases withdrawing). Findings support key elements of the interpersonal process model, showing intra- and interpersonal pathways linking demand/withdraw behavior and emotion and demonstrate the importance of partners' behavioral roles in these linkages.


Accuracy of a predetermined transverse horizontal mandibular axis point.
The transverse horizontal mandibular axis point may be located most precisely by a kinematic process. However, an anatomical method of locating the axis is also an acceptable technique, and an easily determined point that is consistently close to the kinematic axis would simplify transfer of the arc of rotation from the patient to the articulator. This in vivo study compared the location of an anatomically predetermined hinge axis point with the determined kinematic axis. Forty subjects (27 males, 13 females; 23 to 47 years of age) with functionally acceptable occlusion and no detectable clinical signs of temporomandibular disorders participated in the study. The earpiece alignment flags on a mechanical SAM Axiograph III combination flag/face-bow were used to locate the right and left predetermined hinge axis points, 10 mm anterior to the earpiece. The right and left kinematic center of rotation was located as described by Lauritzen and confirmed with the PC Axiotron electronic Axiograph to within 0.25 mm. All points were transferred to 1 mm(2) grid paper on the subject's skin. The distance between each predetermined and kinematic point was measured +/-0.25 mm. Wilcoxon and Mann-Whitney tests were used to examine differences between the left and right axis points and potential significant differences between genders at a significance level of P<.05. The number of occurrences and the distance of the predetermined axis points from the kinematic axis also were described. The mean distance between points was 1.1 mm on the right (range 0.0 to 3.0 mm), 1.2 mm on the left (range 0.0 to 3.0 mm), and 1.1 mm for all 80 points (+/-0.63). More than 96% of the predetermined points were within 2 mm of the kinematic axis, and 67% were within 1 mm. There was no significant difference between the right and left points and no significant differences based on gender. Within the limitations of this study, the results suggest that the predetermined axis point is well within the clinical norm for estimated location of the transverse horizontal mandibular axis.


On the inconsistencies related to prediction of flow into an enclosing hood obstructed by a worker.
The recirculating flow structures formed in the wake of a worker standing in front of an enclosing fume hood were numerically investigated. Two- and three-dimensional, unsteady, laminar/turbulent computations were performed for a Reynolds number (Re) range of 1.0 x 10(3)-1.0 x 10(5). The standard k-epsilon, Renormalization group (RNG) k-epsilon, and Shear Stress Transport (SST) k-omega models were used in Unsteady Reynolds Averaged Navier-Stokes (URANS) computations, and the results were compared with each other and also with the previous predictions reported in the literature. Numerical issues regarding the grid convergence and the inadequacies of turbulence models that may come into play at low Reynolds numbers were addressed. On the whole, SST k-omega model was found to be promising for qualitatively accurate prediction of both steady and unsteady recirculatory flow patterns in the wake of the worker. On the other hand, the standard and RNG k-epsilon models failed in prediction of anticipated unsteadiness at low Reynolds numbers. In a more realistic three-dimensional simulation with SST k-omega model, the anticipated unsteady and recirculating flow field in the wake of the worker was captured. Present results seem to qualitatively agree with the deductions made from experimental analyses in the literature while conflicting with some aspects of the previously reported numerical results. The apparent inconsistencies observed between the current results and those published in the literature were elucidated.


Identification of an immune-based mRNA-lncRNA signature for overall survival in cervical squamous cell carcinoma.
Aim: To better predict the survival of cervical squamous cell carcinoma (CESC) patients, we aimed to construct a signature according to different immune infiltration. Methods: We downloaded the RNA sequences of CESC patients from the Cancer Genome Atlas database. By using single-sample gene set enrichment analysis, we separated the samples into high- and low-immunity groups. Then we separated the samples into training and testing datasets and performed the following analyses: univariate, least absolute shrinkage and selection operator analysis, multivariate Cox regression analyses and weighted gene coexpression network analysis using R software. Gene ontology and Kyoto Encyclopedia of Genes and Genomes studies were performed using the Database for Annotation, Visualization and Integrated Discovery website. Results & conclusion: We finally identified a signature with three mRNAs and two lncRNAs: ADGRG5, HSH2D, ZMAT4, RBAKDN and LINC00200. In short, our study constructed an mRNA-lncRNA signature related to immune infiltration to better predict the survival of CESC patients.


Molecular and functional evolution of the fungal diterpene synthase genes.
Terpenes represent one of the largest and most diversified families of natural compounds and are used in numerous industrial applications. Terpene synthase (TPS) genes originated in bacteria as diterpene synthase (di-TPS) genes. They are also found in plant and fungal genomes. The recent availability of a large number of fungal genomes represents an opportunity to investigate how genes involved in diterpene synthesis were acquired by fungi, and to assess the consequences of this process on the fungal metabolism. In order to investigate the origin of fungal di-TPS, we implemented a search for potential fungal di-TPS genes and identified their presence in several unrelated Ascomycota and Basidiomycota species. The fungal di-TPS phylogenetic tree is function-related but is not associated with the phylogeny based on housekeeping genes. The lack of agreement between fungal and di-TPS-based phylogenies suggests the presence of Horizontal Gene Transfer (HGTs) events. Further evidence for HGT was provided by conservation of synteny of di-TPS and neighbouring genes in distantly related fungi. The results obtained here suggest that fungal di-TPSs originated from an ancient HGT event of a single di-TPS gene from a plant to a fungus in Ascomycota. In fungi, these di-TPSs allowed for the formation of clusters consisting in di-TPS, GGPPS and P450 genes to create functional clusters that were transferred between fungal species, producing diterpenes acting as hormones or toxins, thus affecting fungal development and pathogenicity.


High prevalence of IgG antibodies to Ebola virus in the Efé pygmy population in the Watsa region, Democratic Republic of the Congo.
Factors related to the natural transmission of Ebola virus (EBOV) to humans are still not well defined. Results of previous sero-prevalence studies suggest that circulation of EBOV in human population is common in sub-Saharan Africa. The Efé pygmies living in Democratic Republic of the Congo are known to be exposed to potential risk factors of EBOV infection such as bush meat hunting, entry into caves, and contact with bats. We studied the pygmy population of Watsa region to determine seroprevalence to EBOV infection and possible risks factors. Volunteer participants (N = 300) aged 10 years or above were interviewed about behavior that may constitute risk factors for transmission of EBOV, including exposures to rats, bats, monkeys and entry into caves. Samples of venous blood were collected and tested for IgG antibody against EBOV by enzyme-linked immunosorbent assay (ELISA). The χ2-test and Fisher's exact test were used for the comparison of proportions and the Student's t-test to compare means. The association between age group and anti-EBOV IgG prevalence was analysed by a nonparametric test for trend. The prevalence of anti-EBOV IgG was 18.7 % overall and increased significantly with age (p = 0.023). No association was observed with exposure to risk factors (contacts with rats, bats, monkeys, or entry into caves). The seroprevalence of IgG antibody to EBOV in pygmies in Watsa region is among the highest ever reported, but it remains unclear which exposures might lead to this high infection rate calling for further ecological and behavioural studies.


Apical leakage in canals filled with glass ionomer sealer and gutta-percha after dentin conditioning.
This study assessed in vitro apical leakage in root canals filled with gutta-percha and Ketac-Endo using different dentin conditioning and filling techniques. The root canals of 63 single-rooted teeth were instrumented and randomly divided into the following four groups: group A--canals were irrigated with 25% tannic acid and filled with Ketac-Endo and a standardized gutta-percha cone; group B--canals were irrigated with 1% NaOCl and filled as in group A; group C--canals were irrigated with 25% tannic acid and filled with Ketac-Endo and a chloroform-softened gutta-percha cone adapted to the canal walls; group D--canals were irrigated with 1% NaOCl and filled with Grossman's sealer and laterally condensed gutta-percha. After storage for 72 hours at 37 degrees C and 100% humidity, the gutta-percha was removed to a level 4 mm short of the working length, using a Peeso reamer. Teeth were immersed in India ink for 48 hours and cleared, and the coronal extent of dye penetration examined under a binocular dissecting microscope at x 15 magnification. The mean value of leakage for group A was 0.86 mm, for group B 2.23 mm, for group C 2.34 mm, and for group D 0.56 mm. Tukey's multiple comparison test disclosed significant differences (P < 0.05) between group A and groups B and C, and between group D and groups B and C. Differences between groups A and D, as well as between groups B and C, were not significant.


Assessment of Breast Specimens With or Without Calcifications in Diagnosing Malignant and Atypia for Mammographic Breast Microcalcifications Without Mass: A STARD-Compliant Diagnostic Accuracy Article.
Presence of microcalcifications within the specimens frequently signifies a successful attempt of stereotactic vacuum-assisted breast biopsy (VABB) in obtaining a pathologic diagnosis of the breast microcalcifications. In this study, the authors aimed to assess and compare the accuracy and consistency of calcified or noncalcified specimens obtained from same sites of sampling on isolated microcalcifications without mass in diagnosing high-risk and malignant lesions. To the best of our knowledge, an individual case-based prospective comparison has not been reported.With the approval from institutional review board of our hospital (Chang Gung Memorial Hospital), the authors retrospectively reviewed all clinical cases of stereotactic VABBs on isolated breast microcalcifications without mass from our database. The authors included those having either surgery performed or had clinical follow-up of at least 3 years for analysis. All the obtained specimens with or without calcification were identified using specimen radiographs and separately submitted for pathologic evaluation. The concordance of diagnosis was assessed for both atypia and malignant lesions.A total of 390 stereotactic VABB procedures (1206 calcified and 1456 noncalcified specimens) were collected and reviewed. The consistent rates between calcified and noncalcified specimens were low for atypia and malignant microcalcifications (44.44% in flat epithelial atypia, 46.51% in atypical ductal hyperplasia, 55.73% in ductal carcinoma in situ, and 71.42% in invasive ductal carcinoma). The discordance in VABB diagnoses indicated that 41.33% of malignant lesions would be misdiagnosed by noncalcified specimens. Furthermore, calcified specimens showed higher diagnostic accuracy of breast cancer as compared with the noncalcified specimens (91.54 % versus 69.49%, respectively). The evaluation of both noncalcified specimens and calcified specimens did not show improvement of diagnostic accuracy as compared with evaluating calcified specimens alone (91.54% versus 91.54%, respectively).The high prevalence of diagnostic discordance between the calcified and noncalcified specimens indicated the higher value of calcified specimens in diagnosing atypia and malignant microcalcifications. Noncalcified specimens did not provide additional diagnostic benefit from this study. The separation of calcified and noncalcified specimens may facilitate more focused interpretation from pathologists among the large number of specimens.


Proton magnetic resonance spectroscopy of the brain during acute hypoxia-ischemia and delayed cerebral energy failure in the newborn piglet.
Studies of the brains of severely birth-asphyxiated infants using proton (1H) magnetic resonance spectroscopy (MRS) have shown changes indicating a rise in cerebral lactate (Lac) and a fall in N-acetylaspartate (Naa). The aim of this study was to test two hypotheses: 1) that these changes can be reproduced in the newborn piglet after transient reversed cerebral hypoxiaischemia, and their time course determined; and 2) that changes in Lac peak-area ratios are related to changes in phosphorylation potential as determined by phosphorus (31P) MRS. Eighteen piglets aged < 24 h were anesthetized and ventilated. Twelve underwent temporary occlusion of the carotid arteries and hypoxemia, and six served as sham-operated controls. 1H and 31P spectra were acquired alternately, both during the insult and for the next 48 h, using a 7-tesla spectrometer. During hypoxiaischemia, the median Lac/total creatine (Cr) peak-area ratio rose from a baseline of 0.14 (interquartile range 0.07-0.27), to a maximum of 4.34 (3.33-7.45). After resuscitation, Lac/Cr fell to 0.75 (0.45-1.64) by 2 h, and then increased again to 2.43 (1.13-3.08) by 48 h. At all stages after resuscitation Lac/Cr remained significantly above baseline and control values. Naa/Cr was significantly reduced below baseline and control values by 48 h after resuscitation. The increases in the Lac peak-area ratios were concomitant with the falls in the [phosphocreatine (PCr)*]/ [inorganic phosphate (Pi)] ratio, during both acute hypoxiaischemia and delayed energy failure. The maximum Lac/Naa during delayed energy failure correlated strongly with the minimum [nucleotide triphosphate (NTP)]/[exchangeable phosphate pool (EPP)] (r = -0.94, p < 0.0001). We conclude that both hypotheses have been confirmed.


Analysis of the relationship between sex and chromosomal aberrations in colorectal cancer by comparative genomic hybridization.
Colorectal cancer is thought to be more common in men than in women. The chromosomal locations of DNA gains and losses in surgical specimens of colorectal tumours were detected by comparative genomic hybridization and were compared by gender. Five chromosomal regions, 7p, 8p, 8q, Xp and Xq, contained multiple gains that were significantly more common in males than in females, and within these regions, the differences were significant for Xp21, Xp11.3, Xp11.4 and Xq26. Regions 1p, 3q, 11q, 12p, 12q and 15q contained multiple sites of gain that were significantly more common in females than in males. Tumours from male and female patients showed significantly more losses at 11p and 15q, and at 4q and Xq, respectively. The fact that gains in X-chromosomal regions were detected with a significantly higher frequency in tumours from male patients suggests that the difference between the genders might be explained by X-chromosomal inactivation.


Is there a relationship between fall status, cognition and cerebellar lobule volume in patients with multiple sclerosis?
In this prospective case control study, relationship of detailed cerebellar volumetric data and cognition in patients with multiple sclerosis considering falling status using 3 D MRI and network analysis were evaluated. Participants consist of 106 adults with relapsing-remitting multiple sclerosis. Scores of Montreal cognitive assessment test, symbol digit modality Test, nine-hole peg test, berg balance scale test, timed up and go test, timed 25-foot walk test were worse in faller group than non faller group (p < 0.05 for all tests). There was no significant difference in terms of cerebellar lobule volumes between groups. But using artificial intelligence (AI) based network analysis, we brought a new perspective to interpreting the relationship between the cerebellum, cognition, gait, and balance. Overall, data from the study suggest a possible relationship between cerebellar volume changes and cognitive dysfunction through connectivity analysis in patients with multiple sclerosis. Further studies are needed to examine this issue by using connectivity analysis.


Nonemergency use of slow infusions of diazoxide.
Slow infusions of diazoxide were administered to 10 hypertensive patients who had stable, nonaccelerated hypertension. The 10-min diazoxide infusion was associated with a 16% average reduction in arterial pressure, a 21% average increase in heart rate, a 16% average increase in cardiac output, and a 36% reduction in total peripheral resistance. These changes in hemodynamic parameters lasted for an average of 70 min. It was concluded that slow infusions of diazoxide produce a consistent and predictable antihypertensive effect in patients with stable, nonaccelerated hypertension. Slow infusions of diazoxide may have a limited use in nonemergency situations where an abrupt change in arterial pressure is to be avoided and a parenteral antihypertensive agent is needed.


Application of pyrosequencing method for investigating the diversity of synechococcus subcluster 5.1 in open ocean.
Synechococcus are distributed throughout the world's oceans and are composed of diverse genetic lineages. However, as they are much less abundant than Prochlorococcus in oligotrophic open oceans, their in-depth genetic diversity cannot be investigated using commonly used primers targeting both Prochlorococcus and Synechococcus. Thus, in this study, we designed a primer specific to the 16S-23S rRNA internal transcribed spacer (ITS) of the Synechococcus subcluster 5.1. Using the primer, we could selectively amplify Synechococcus sequences in oligotrophic seawater samples. Further, we showed that a barcoded amplicon pyrosequencing method could be applicable to investigate Synechococcus diversity using sequences retrieved in GenBank and obtained from environmental samples. Allowing sequence analyses of a large number of samples, this high-throughput method would be useful to study global biodiversity and biogeographic patterns of Synechococcus in marine environments.


A Computational Study of the Electronic Energy and Charge Transfer Rates and Pathways in the Tetraphenyldibenzoperiflanthene/Fullerene Interfacial Dyad.
The electronic transition rates and pathways underlying interfacial charge separation in tetraphenyldibenzoperiflanthene:fullerene (DBP:C70) blends are investigated computationally. The analysis is based on a polarization-consistent framework employing screened range-separated hybrid functional in a polarizable continuum model to parametrize Fermi's golden rule rate theory. The model considers the possible transitions within the 25 lowest excited states of a DBP:C70 dyad that are accessible by photoexcitation. The different identified pathways contributing to charge carrier generation include electron and hole transfer and backtransfer, exciton transfer, and internal relaxation steps. The larger density of states of C70 appears to explain the previously observed larger efficiency for charge separation through hole transfer mechanism. We also analyze the validity of the high-temperature and short-time semiclassical approximations of the FGR theory, where both overestimated and underestimated Marcus theory based constants can be affected.


Emerging roles of miR-210 and other non-coding RNAs in the hypoxic response.
Hypoxia is a key component of the tumor microenvironment and represents a well-documented source of therapeutic failure in clinical oncology. Recent work has provided support for the idea that non-coding RNAs, and in particular, microRNAs, may play important roles in the adaptive response to low oxygen in tumors. Specifically, all published studies agree that the induction of microRNA-210 (miR-210) is a consistent feature of the hypoxic response in both normal and malignant cells. miR-210 is a robust target of hypoxia-inducible factors, and its overexpression has been detected in a variety of diseases with a hypoxic component, including most solid tumors. High levels of miR-210 have been linked to an in vivo hypoxic signature and to adverse prognosis in breast and pancreatic cancer patients. A wide variety of miR-210 targets have been identified, pointing to roles in mitochondrial metabolism, angiogenesis, DNA damage response, apoptosis, and cell survival. Such targets are suspected to affect the development of tumors in multiple ways; therefore, an increased knowledge about miR-210's functions may lead to novel diagnostic and therapeutic approaches in cancer.


Typed Operation Notes in Rural Western Australia: Improving Patient Care.
Given the geographical area of the Kimberley region in Western Australia (WA) and the nomadic nature of its residents and medical staff, widespread access and clarity of surgical clinical information are necessary to provide accurate and timely post-surgical care. The aims of this project were: to evaluate the quality of operation notes and secondly, to evaluate multidisciplinary staff perceptions of the impact of the introduction of typed operation notes for general surgery in the Kimberley region from 2019 to 2020. The quality of 100 general surgery operation notes (50 typed and 50 handwritten) were reviewed against the Royal College of Surgeons England (RCSEng) operation note guidelines. Cases were selected at random and reviewed by a resident medical officer. Multidisciplinary staff perceptions of communication were assessed through an anonymous electronic survey across emergency departments, general practices, nursing staff, and allied health staff members from the top three population centres: Broome, Derby and Kununurra. Typed operation notes with pre-loaded data (date, time, etc.) and mandatory fields (surgical count correct, etc.) increased recorded information and quality of content by 45% when compared to handwritten notes. When compared to RCSEng standards for free text, anticipated blood loss (one typed note) and abbreviation use (44 typed and 37 handwritten) showed ongoing user-dependent areas for improvement. A review of multidisciplinary staff perceptions (79 returned surveys) showed 60% of handwritten notes were seen to have a negative impact on timely post-operative care. Overall, typed notes increased legibility with a perceived improvement in acronyms/abbreviations and completeness of documentation. More than 90% of staff suggested an extension of typed notes for all surgical operations would be beneficial. The Kimberley region poses a unique set of challenges to the continuity of post-surgical care. This review shows typed operation notes improve legibility (100%) and increased congruence with established guidelines by 45%. It also shows a successful model for increased local and metropolitan multidisciplinary access across remote WA for timely post-operative care. In an unprecedented time where elective surgical procedures are being reduced to meet pandemic demands, now is the time to review, consider and institute practices that improve intra-operative and post-operative care.


Engineering Soluble Diketopyrrolopyrrole Chromophore Stacks from a Series of Pd(II)-Based Ravels.
A strategy to engineer the stacking of diketopyrrolopyrrole (DPP) dyes based on non-statistical metallosupramolecular self-assembly is introduced. For this, the DPP backbone is equipped with nitrogen-based donors that allow for different discrete assemblies to be formed upon the addition of Pd(II), distinguished by the number of π-stacked chromophores. A Pd3 L6 three-ring, a heteroleptic Pd2 L2 L'2 ravel composed of two crossing DPPs (flanked by two carbazoles), and two unprecedented self-penetrated motifs (a Pd2 L3 triple and a Pd2 L4 quadruple stack), were obtained and systematically investigated. With increasing counts of stacked chromophores, UV/Vis absorptions red-shift and emission intensities decrease, except for compound Pd2 L2 L'2 , which stands out with an exceptional photoluminescence quantum yield of 51 %. This is extraordinary for open-shell metal containing assemblies and explainable by an intra-assembly FRET process. The modular design and synthesis of soluble multi-chromophore building blocks offers the potential for the preparation of nanodevices and materials with applications in sensing, photo-redox catalysis and optics.


Evaluation of collaborative mental health services in Latin America: Theoretical and methodological basis.
Approaches to collaborative mental health care (CMHC) have been implemented in many countries to strengthen the accessibility and delivery of mental health services in primary care. However, there are not well-defined frameworks to evaluate CMHC models. The purpose of this article is to identify, contextualize and discuss relevant health services research approaches, theory, and evaluation models for the development of an appropriate evaluation framework in order to foster effective CMHC in Latin America. A comprehensive literature review informed a critical analysis of relevant theories and alternative methods to be considered in the development of the framework. Specific health services research frameworks are discussed in the context of evaluating CMHC. Two theoretical perspectives - collaboration theory and systems theory - and three evaluation models- realistic, developmental and collaborative - are analyzed in terms of their relevance. Methodological implications are identified. An appropriate evaluation framework for CMHC in Latin America needs to reflect theoretical and contextual considerations and relevant evaluation approaches and methods, including key dimensions and attributes/variables, core indicators, and recommendations for implementation.


Peroxynitrite increases the degradation of aconitase and other cellular proteins by proteasome.
We report that exposure of aconitase to moderate concentrations of peroxynitrite, 3-morpholinosydnonimine (SIN-1; a superoxide- and nitric oxide-liberating substance), or hydrogen peroxide, inhibits the enzyme and enhances susceptibility to proteolytic digestion by the isolated 20 S proteasome. Exposure to more severe levels of oxidative stress, from these same agents, causes further inhibition of the enzymatic activity of aconitase but actually decreases its proteolytic breakdown by proteasome. It should be noted that the superoxide and nitric oxide liberated by SIN-1 decomposition react to form a steady flux of peroxynitrite. S-Nitroso-N-acetylpenicillamine, a compound that liberates nitric oxide alone, causes only a small loss of aconitase activity (25% or less) and has no effect on the proteolytic susceptibility of the enzyme. Proteasome also seems to be the main protease in cell lysates that can degrade aconitase after it has been oxidatively modified by exposure to peroxynitrite, SIN-1, or hydrogen peroxide. Using cell lysates isolated from K562 cells treated for several days with an antisense oligodeoxynucleotide to the initiation codon region of the C2 subunit of proteasome (a treatment which diminishes proteasome activity by 50-60%), the enhanced degradation of moderately damaged aconitase was essentially abolished. Other model proteins as well as complex mixtures of proteins, such as cell lysates, also exhibit enhanced proteolytic susceptibility after moderate SIN-1 treatment. Therefore we conclude that peroxynitrite reacts readily with proteins and that mild modification by peroxynitrite results in selective recognition and degradation by proteasome.


Characterization and Prediction of Anxiety in Adolescents with Autism Spectrum Disorder: A Longitudinal Study.
Anxiety is one of the most common comorbidities in youth with autism spectrum disorder (ASD). The current study's aims were: To examine the frequency of elevated anxiety symptoms in adolescents diagnosed with ASD in toddlerhood; To explore the impact of comorbid anxiety in adolescents on clinical presentation; To evaluate variables in toddlerhood that associate with anxiety symptom severity in adolescence. The study included 61 adolescents (mean age = 13:8y) diagnosed with ASD in toddlerhood (T1). Participants underwent a comprehensive assessment of cognitive ability, adaptive skills and autism severity at T1 and again as adolescents (T2), and an evaluation of anxiety symptoms at T2. For the first aim, the most prevalent anxiety subtypes noted in adolescence were separation (39.3%), social (27.9%) and generalized anxiety (18.0%). For the second aim, cognitive ability, autism severity and adaptive skills in adolescents with and without elevated anxiety symptoms scores of any type did not differ significantly. For the third aim, younger age at adolescence was associated with more severe separation and generalized anxiety symptoms. Higher cognitive ability and adaptive skills in toddlerhood were associated with elevated generalized anxiety symptoms in adolescence. Lower adaptive behaviors and repetitive behaviors (RRBs) correlated with elevated social anxiety symptoms. Lower cognitive abilities and more severe RRBs in toddlerhood predicted separation anxiety in adolescence. The study sheds light on early characteristics in ASD that associate with anxiety symptom severity in adolescence. The type of elevated anxiety symptoms presented in adolescence associated with the level of cognitive ability, adaptive skills and RRBs in toddlerhood.


Valvular osteoclasts in calcification and aortic valve stenosis severity.
Bone remodeling in calcified aortic valves is thought to originate from microfractures at multiple sites of the valve, at which osteoclasts and osteoblasts are recruited. The aim of the present study was to assess circulating mediators of bone homeostasis, correlate them to the severity of stenosis and explore the spatio-temporal distribution of bone turnover in different parts of calcified aortic valve tissue. Plasma and explanted aortic valves were obtained from 46 patients undergoing aortic valve replacement surgery. Plasma levels of tartrate-resistant acid phosphatase (TRAP), receptor activator of nuclear-κB (RANK) ligand and Runt-related transcription factor 2 (Runx2/Cbfa1) exhibited a significant correlation to the severity of aortic stenosis. mRNA levels in normal, thickened and calcified parts of aortic valves assessed by quantitative real-time PCR were significantly elevated in calcified parts of valves for TRAP (5.08 ± 1.6-fold, P<0.001) RANK ligand (8.6 ± 4.2-fold, P<0.001) and RANK (1.98 ± 0.78-fold, P=0.015). In an age, gender and aortic valve anatomy-adjusted multivariable regression analysis the local transcript levels of TRAP correlated significantly with echocardiographic parameters quantifying stenosis severity in early stages, whereas the expression level of Runx2/Cbfa1 was a predictor of the stenosis severity in advanced stages. These findings suggest a critical role of bone turnover as a determinant of aortic stenosis severity.


Recent advances in metacommunities and meta-ecosystem theories.
Metacommunity theory has provided many insights into the general problem of local versus regional control of species diversity and relative abundance. The metacommunity framework has been extended from competitive interactions to whole food webs that can be described as spatial networks of interaction networks. Trophic metacommunity theory greatly contributed to resolving the community complexity-stability debate by predicting its dependence on the regional spatial context. The meta-ecosystem framework has since been suggested as a useful simplification of complex ecosystems to apply this spatial context to spatial flows of both individuals and matter. Reviewing the recent literature on metacommunity and meta-ecosystem theories suggests the importance of unifying theories of interaction strength into a meta-ecosystem framework that captures how the strength of spatial, species, and ecosystem fluxes are distributed across location and trophic levels. Such integration predicts important feedback between local and regional processes that drive the assembly of species, the stability of community, and the emergence of ecosystem functions, from limited spatial fluxes of individuals and (in)organic matter. These predictions are often mediated by the maintenance of environmental or endogenous fluctuations from local to regional scales that create important challenges and opportunities for the validation of metacommunity and meta-ecosystem theories and their application to conservation.


Diazeniumdiolate ions as leaving groups in anomeric displacement reactions: a protection-deprotection strategy for ionic diazeniumdiolates.
Diazeniumdiolate ions [R2N-N(O)=N-O-] are of growing interest pharmacologically for their ability to generate up to two molar equivalents of bioactive nitric oxide (NO) spontaneously on protonating the amino nitrogen. Accordingly, their stability increases as the pH is raised. Here we show that the corresponding beta-glucosides [R2N-N(O)=N-O-Glc] decreased in stability with pH; when R2N was diethylamino, the rate equation was kobs = ko + kOH- [OH-], where ko = 7.8 x 10-7 s-1 and kOH- = 5.3 x 10-3 M-1 s-1. The primary products were 1,6-anhydroglucose and the regenerated R2N-N(O)=N-O- ion. The results were qualitatively similar to those of beta-glucosyl fluoride and p-nitrophenoxide, whose hydrolyses have been rationalized as proceeding via a glycal oxide intermediate. This chemistry offers a convenient strategy for protecting heat- and acid-sensitive diazeniumdiolate ions during manipulations that would otherwise destroy them. As an example, a poly(urethane) film that generated NO in physiological buffer at a surface flux comparable to that of the mammalian vascular endothelium was prepared by glucosylating the ionic diazeniumdiolate group attached to the diol monomer before reacting it with the bis-isocyanate, then removing the saccharide with base when the protecting group was no longer needed.


The potential of organoids in urological cancer research.
Technical advances in the development of organoid systems enable cell lines, primary adult cells, or stem or progenitor cells to develop into diverse, multicellular entities, which can self-renew, self-organize, and differentiate. These 3D organoid cultures have proven to be of value in increasing our understanding of the biology of disease and offer the potential of regenerative and genetic therapies. The successful application of 3D organoids derived from adult tissue into urological cancer research can further our understanding of these diseases and could also provide preclinical cancer models to realize the precision medicine paradigm by therapeutic screening of individual patient samples ex vivo. Kidney organoids derived from induced pluripotent stem cells provide personalized biomarkers, which can be correlated with genetic and clinical information. Organoid models can also improve our comprehension of aspects of particular diseases; for example, in prostate cancer, 3D organoids can aid in the identification of tumour-initiating cells from an epithelial cell lineage. Furthermore, kidney organoid differentiation from human pluripotent stem cells enables gene editing to model disease in kidney tubular epithelial cells. State-of-the-art human organoid cultures have potential as tools in basic and clinical research in renal, bladder, and prostatic diseases.


Evaluation of diffusion-weighted MR imaging at inclusion in an active surveillance protocol for low-risk prostate cancer.
We aimed to determine whether diffusion-weighted magnetic resonance imaging, by means of the apparent diffusion coefficient (ADC), is able to guide magnetic resonance-guided biopsy in patients fit for active surveillance (AS) and identify patients harboring high-grade Gleason components not suitable for AS. Our study was approved by the institutional review board of all participating hospitals, and all patients signed informed consent at inclusion. Fifty-four consecutive patients with low-risk prostate cancer (PCa) underwent multiparametric magnetic resonance imaging (MP-MRI) at inclusion for AS. Cancer-suspicious regions (CSRs) upon 3-T MP-MRI were identified in all patients, and magnetic resonance-guided biopsy was performed in all CSRs to obtain histopathological verification. For all CSRs, a median ADC (mADC) was calculated. Wilcoxon signed ranks and Mann-Whitney tests was performed to detect differences between the groups. We used the area under the receiver operating characteristic curve to evaluate the accuracy of mADC to predict the presence of PCa in a CSR. Level of statistical significance was set at P < 0.05. Mean mADC in the CSRs with PCa was 1.04 × 10⁻³ mm²/s (SD, 0.29), whereas the CSRs with no PCa displayed a mean mADC of 1.26 × 10⁻³ mm²/s (SD, 0.25; P < 0.001). Cancer-suspicious regions with a high-grade Gleason component displayed a mean mADC of 0.84 × 10⁻³ mm²/s (SD, 0.35) vs a mean mADC for the low-grade CSRs of 1.09 × 10⁻³ mm²/s (SD, 0.25; P < 0.05). A diagnostic accuracy of mADC for predicting the presence of PCa in a CSR with an area under the receiver operating characteristic curve of 0.73 was established (95% confidence interval, 0.61-0.84). Median ADC is able to predict the presence and grade of PCa in CSRs identified by MP-MRI.


Concurrent validation of the Actigraph gt3x+, Polar Active accelerometer, Omron HJ-720 and Yamax Digiwalker SW-701 pedometer step counts in lab-based and free-living settings.
Activity monitors are frequently used to assess activity in many settings. But as technology advances, so do the mechanisms used to estimate activity causing a continuous need to validate newly developed monitors. The purpose of this study was to examine the step count validity of the Yamax Digiwalker SW-701 pedometer (YX), Omron HJ-720 T pedometer (OP), Polar Active accelerometer (PAC) and Actigraph gt3x+ accelerometer (AG) under controlled and free-living conditions. Participants completed five stages of treadmill walking (n = 43) and a subset of these completed a 3-day free-living wear period (n = 37). Manually counted (MC) steps provided a criterion measure for treadmill walking, whereas the comparative measure during free-living was the YX. During treadmill walking, the OP was the most accurate monitor across all speeds (±1.1% of MC steps), while the PAC underestimated steps by 6.7-16.0% per stage. During free-living, the OP and AG counted 97.5% and 98.5% of YX steps, respectively. The PAC overestimated steps by 44.0%, or 5,265 steps per day. The Omron pedometer seems to provide the most reliable and valid estimate of steps taken, as it was the best performer under lab-based conditions and provided comparable results to the YX in free-living. Future studies should consider these monitors in additional populations and settings.


Anthracycline-induced cardiotoxicity in children with malignancies.
The anthracyclines (ATC) have been used in the treatment of many pediatric patients with malignancies. Their use, however, has been limited by a serious and potentially irreversible cardiotoxicity. The medical records of pediatric patients with malignancies who received ATC at the Department of Pediatrics, Kyushu University, from January 1985 to December 1994 were reviewed. Among the 120 children with malignancies who received ATC, six patients (5%) developed congestive heart failure, 33 (28%) had subclinical cardiac dysfunction, whereas 81 (66%) showed no evidence of cardiac dysfunction. The incidence of ATC cardiotoxicity increased at higher cumulative doses. The earliest cardiotoxicity, however, was observed in a patient who received 180 mg/m2 unassociated with other risk factors, showing the marked individual variations in its development. Children younger than 4 years had a higher incidence of cardiotoxicity compared with those older than 4 years (P < 0.01) and are therefore more vulnerable to ATC cardiotoxicity. Because serial myocardial biopsies and stress testing are not practical in younger children, and radionuclide angiogram is rather costly and not appropriate for frequent evaluation, non-invasive tests such as echocardiography and electrocardiography (ECG) still play the main role for serial cardiac monitoring in these age groups. Radionuclide angiogram should be added as the confirmatory test modality.


Processing of naked 45-S ribosomal RNA precursor in vitro by an RNA-associated endoribonuclease.
A processing endoribonuclease was isolated from the cytoplasm of chick embryos. The enzyme was easily obtained using an RNA extraction procedure based on a mild deproteinization with Sarkosyl and cold phenol/chloroform. This technique assured the recovery of several proteins and the endoribonuclease in association with the RNA. It was demonstrated that this endoribonuclease was capable of promoting, in vitro, a precise processing of naked 45-S ribosomal RNA precursor to molecules resembling the intermediates as well as the 28-S and 18-S cytoplasmic RNAs found in vivo. The presence of magnesium ions was required for the correct processing function of the enzyme. In addition, under the same conditions, the mature ribosomal RNA substrates were degraded at a slower rate by this RNA-associated RNase. It was possible to fractionate the enzymatic preparation into two different populations by means of a sucrose gradient: one associated and the other partially free of an RNA component. The effect of the intrinsic RNA associated with the endoribonuclease on the enzymatic activity was tested by analyzing both the enzymatic populations and the total enzymatic preparation treated with pronase or with immobilized pancreatic RNase. In all cases in which the RNA component was present, the enzyme showed processing activity. On the other hand, when the RNA component was absent or at least partially degraded the enzyme proved to be more active in processing precursor molecules and in promoting extensive degradation of mature RNA species. Although the presence of RNA in association with the enzyme was demonstrated, its role in the regulation of the enzymatic activity is yet not clear.


The plastic surgical adhesive drape: an evaluation of its efficacy as a microbial barrier.
A microbial evaluation was made of adhesive plastic surgical drapes and cloth surgical drapes. These studies were done both during surgery and in the laboratory. The plastic drape does not allow bacterial penetration, lateral migration does not occur, skin bacteria do not multiply under the drape within the time periods studied and the patient drapes are held in place with their use. When wet, cloth drapes showed profuse bacterial penetration. Dry cloth showed less bacterial penetration as compared to wet cloth. Lateral migration under cloth drapes was not possible to assess due to a high level of penetration. The surface of cloth showed a higher level of bacterial contamination during the surgical procedures. Deep wound cultures collected just prior to closing showed 60% contamination when cloth was used compared to 6% when plastic was employed. The micro-organisms recovered from the various sites sampled were identified. Finally, in addition to the positive aseptic benefits afforded by plastic adhesive drapes, aesthetic features such as a more delineated operative field and elimination of towel clips make this product a useful adjunct to the surgeon's armamentarium.


Familial aggregation of cluster headache.
Several studies suggest a strong familial aggregation for cluster headache (CH), but so far none of them have included subjects with probable cluster headache (PCH) in accordance with the International Classification of Headache Disorders. To identify cases of probable cluster headache and to assess the familial aggregation of cluster headache by including these subjects. Thirty-six patients attending a headache consultation and diagnosed with trigeminal autonomic headaches were subjected to a questionnaire-based interview. A telephone interview was also applied to all the relatives who were pointed out as possibly affected as well as to some of the remaining relatives. Twenty-four probands fulfilled the criteria for CH or PCH; they had 142 first-degree relatives, of whom five were found to have CH or PCH, including one case of CH sine headache. The risk for first-degree relatives was observed to be increased by 35- to 46-fold. Our results suggest a familial aggregation of cluster headache in the Portuguese population.


Malignant tumours of the stomach.
The commonest malignant tumour of the stomach is gastric carcinoma. Although the incidence of this neoplasm has declined worldwide, it still ranks fifth to seventh as a cause of cancer-related deaths in the West where the overall 5-year survival from the disease remains poor and averages 5-10%. The decline in the incidence of gastric cancer during the past 4 decades is largely confined to antral tumours of the intestinal type. During this period an absolute increase in the prevalence of cancers of the upper third of the stomach has been observed. Japan is the only country where the mortality has declined proportionately more than the decreased incidence of the disease. This is attributed to a successful screening programme for the detection of early gastric cancer, better staging and probably more aggressive surgical treatment with an extended lymphadenectomy. The latter remains unproven and is currently being evaluated prospectively in a Medical Research Council trial being conducted in the United Kingdom. Surgical excision remains the mainstay of therapy. To date there has been no firm evidence that peri-operative radiotherapy imparts any therapeutic advantage. Overall survival is not improved by adjuvant combination chemotherapy although the early results of a modified FAM regimen with cisplatin are encouraging in terms of response rates. Gastric carcinoids most frequently arise in patients with pernicious anaemia due to the prolonged hypergastrinaemia. Although they can metastasize, their prognosis is good and resection is indicated even in the presence of deposits. Gastric lymphoma accounts for 0.5-3% of gastric malignancies and is the prototype of malignant lymphomas arising from mucosa associated lymphoid tissue (MALT) present in the gastrointestinal tract and other organs. The majority of these tumours are B-cell lymphomas which are slow growing and tend to remain localized until late in the course of the disease. The various accepted histological classifications are difficult to apply to MALT lymphomas which are nowadays regarded as a distinct entity the prognosis of which is determined more by stage than histological type. They frequently cause diagnostic problems because of their indolent growth characteristics and their close simulation to benign lymphoid infiltrates. In the past, the term "pseudolymphoma" was applied to those extranodal proliferations of lymphoid tissue the nature of which was uncertain. This problem has been resolved by the advent of immunocytochemical and molecular biological techniques such that it is now always possible to distinguish a reactive from a neoplastic lymphoid infiltrate.(ABSTRACT TRUNCATED AT 400 WORDS)


Regulation of Na+ channels by luminal Na+ in rat cortical collecting tubule.
1. The idea that luminal Na+ can regulate epithelial Na+ channels was tested in the cortical collecting tubule of the rat using whole-cell and single-channel recordings. Here we report results consistent with the idea of Na+ self-inhibition. 2. Macroscopic amiloride-sensitive currents (INa) were measured by conventional whole-cell clamp. INa was a saturable function of external Na+ concentration ([Na+]o) with an apparent Km of 9 mM. Single channel currents (iNa) were measured in cell-attached patches. iNa increased with pipette Na+ concentration with an apparent Km of 48 mM. Since INa = (iNa)NPo, the different Km values imply that the channel density (N) and/or open probability (Po) increase as [Na+]o decreases. Reduction of [Na+]o after increasing intracellular Na+ concentration also increased the outward amiloride-sensitive conductance, consistent with activation of the Na+ channels. 3. The underlying mechanism was studied by changing pipette Na+ concentration while recording from cell-attached patches. No increase in NPo was observed, suggesting that the effect is not a direct interaction between [Na+]o and the channel. 4. [Na+]o was varied outside the patch-clamp pipette while recording from cell-attached patches. When amiloride was in the bath to prevent Na+ entry, no change in NPo was observed. 5. Activation of the channels by hyperpolarization was observed with 140 mM Na+o but not with 14 mM Na+o. 6. The results are consistent with the concept of self-inhibition of Na+ channels by luminal Na+. Activation of the channels by lowering [Na+]o is not additive with that achieved by hyperpolarization.


Alpha-fetoprotein and albumin synthesis by heterotransplanted rat liver tumor cells.
Epithelial cells, isolated from the hepatic tissue of rats bearing carcinogen-induced neoplastic liver nodules, were analyzed by immunological methods for the production of specific liver proteins during alternate in vitro-in vivo passage. Heterotransplantation of several long-term rat liver cell lines into nude mice and subsequent re-establishment in vitro resulted in the derivation of hepatocyte cultures expressing both alpha-fetoprotein and albumin. Prior to passage in vivo these cell lines synthesized by one or neither of these proteins in vitro. Alternate in vitro-in vivo passage thus appears to potentiate tissue-specific protein expression by established liver cell lines.


Formation of zinc phosphate polymers and networks through the insertion of metathiophosphates into zinc dialkyldithiophosphates.
The interactions between metathiophosphates (MTPs) and zinc dialkyldithiophosphates (ZnDDPs) were explored through molecular simulation. The results highlight a novel reaction leading to the formation of zinc phosphate polymers and networks through the insertion of MTPs into ZnDDPs.


Amide modes of the alpha-helix: Raman spectroscopy of filamentous virus fd containing peptide 13C and 2H labels in coat protein subunits.
The filamentous virus fd consists of a single-stranded DNA genome sheathed by 2700 copies of a 50-residue alpha-helical subunit (protein pVIII) and serves as a model assembly of alpha-helices. To advance vibrational assignments for the alpha-helix, we have investigated Raman spectra of fd virions containing 13C and 2H (deuterium) labels at various main-chain sites of the pVIII subunits. 13C was introduced at specific peptide carbonyls, while deuterium was introduced at selected alpha-carbon (Calpha) and amide nitrogen positions. Interpretation of the Raman spectra reveals a previously unrecognized alpha-helix band in the spectral interval 730-745 cm-1, tentatively assigned to a carbonyl in-plane bending mode (amide IV). Experimental evidence has also been obtained for a distinctive alpha-helix marker near 1345 cm-1, assigned to a coupled Calpha-H bending and Calpha-C stretching mode. The fd virions containing 13C-labeled carbonyls exhibit unexpectedly complex amide I profiles, consisting of multiple band components. Amide I splitting resulting from 13C substitution of carbonyls is attributed to decoupling of transition-dipole interactions normally occurring in the extended pVIII helix. The present study identifies novel conformation-dependent Raman bands in a native alpha-helix assembly, confirms amide I and amide III assignments proposed previously for filamentous viruses, and facilitates new Raman assignments for the packaged ssDNA. The alpha-helix markers identified here should also be useful in conformation analyses of other proteins by Raman spectroscopy.


CO2 -dependent metabolic modulation in red blood cells stored under anaerobic conditions.
Anaerobic red blood cell (RBC) storage reduces oxidative damage, maintains adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (DPG) levels, and has superior 24-hour recovery at 6 weeks compared to standard storage. This study will determine if removal of CO2 during O2 depletion by gas exchange may affect RBCs during anaerobic storage. This is a matched three-arm study (n = 14): control, O2 and CO2 depleted with Ar (AN), and O2 depleted with 95%Ar/5%CO2 (AN[CO2 ]). RBCs in additives AS-3 or OFAS-3 were evenly divided into three bags, and anaerobic conditions were established by gas exchange. Bags were stored at 1 to 6°C in closed chambers under anaerobic conditions or ambient air, sampled weekly for up to 9 weeks for a panel of in vitro tests. A full metabolomics screening was conducted for the first 4 weeks of storage. Purging with Ar (AN) results in alkalization of the RBC and increased glucose consumption. The addition of 5% CO2 to the purging gas prevented CO2 loss with an equivalent starting and final pH and lactate to control bags (p > 0.5, Days 0-21). ATP levels are higher in AN[CO2 ] (p < 0.0001). DPG was maintained beyond 2 weeks in the AN arm (p < 0.0001). Surprisingly, DPG was lost at the same rate in both control and AN[CO2 ] arms (p = 0.6). Maintenance of ATP in the AN[CO2 ] arm demonstrates that ATP production is not solely a function of the pH effect on glycolysis. CO2 in anaerobic storage prevented the maintenance of DPG, and DPG production appears to be pH dependent. CO2 as well as O2 depletion provides metabolic advantage for stored RBCs.


Laryngeal cysts in adults: a clinicopathologic study of 20 cases.
Clinically significant cysts in the region of the larynx are uncommon. In this study, 20 cases were recovered for the period 1969-1983. There were eight males and 12 females whose ages ranged from 28-76 years and whose most common symptom combinations were hoarseness (9), local foreign body sensation (5), and pain (4). Laryngoscopic examination revealed cystic lesions 0.5-3.0 cm. in diameter, all localized to the supraglottis with half in the vallecula . Histologically, 17 cysts were closely associated with tonsillar lymphoid tissue. Nine cysts, having squamous lined crypt-like structures and abundant follicular lymphoid tissue, were designated "tonsillar cysts." Eight cysts had a prominent epithelial component and focal follicular lymphoid aggregates. Three cysts were unassociated with lymphoid tissue. Two of these, located in the ventricles of elderly patients, were oncocytic cysts. Both showed oncocytic changes on the surface mucosa and in adjacent salivary gland tissue. In one case, a biopsy of the contralateral ventricle demonstrated oncocytic metaplasia. Treatment in all cases was by surgical excision. Follow-up on 13 patients ranging from three months to 9 years revealed one recurrence of an oncocytic cyst one year after initial removal. We conclude that follicular lymphoid tissue is present in most laryngeal cysts and may play some pathogenetic role in their formation. Furthermore, oncocytic cysts are different from other laryngeal cysts, having a characteristic location, age group, field effect, and recurrence rate.


The etiology of Mycobacterium avium subspecies paratuberculosis in Crohn's disease.
The etiology of Crohn's disease (CD) has drawn heated controversy in the literature. Compelling evidence in the literature has accumulated lately that could incriminate the Mycobacterium avium subspecies paratuberculosis (MAP), the well-known agent of John's disease in cattle. The evidence is isolation of the organism or its DNA and RNA, detection of the anti-MAP antibodies in Crohn's patients, increasing incidents of CD in areas close to the cattle pastures, and the possibility of treating the disease with the antibiotics. The group that favors the immune dysregulation theory considered this evidence circumstantial due to the variations in these reports. The treatment of CD with humanized anti-tumor necrosis factor-alpha antibodies is considered great endorsement to the immune dysregulation theory. The endless debate could jeopardize public health rather than bring a final solution. Reconciliation between the 2 theories appears inevitable in view of possible classification of this disease as a zoonotic.


Prevalence and correlates of helminth co-infection in Kenyan HIV-1 infected adults.
Deworming HIV-1 infected individuals may delay HIV-1 disease progression. It is important to determine the prevalence and correlates of HIV-1/helminth co-infection in helminth-endemic areas. HIV-1 infected individuals (CD4>250 cells/ul) were screened for helminth infection at ten sites in Kenya. Prevalence and correlates of helminth infection were determined. A subset of individuals with soil-transmitted helminth infection was re-evaluated 12 weeks following albendazole therapy. Of 1,541 HIV-1 seropositive individuals screened, 298 (19.3%) had detectable helminth infections. Among individuals with helminth infection, hookworm species were the most prevalent (56.3%), followed by Ascaris lumbricoides (17.1%), Trichuris trichiura (8.7%), Schistosoma mansoni (7.1%), and Strongyloides stercoralis (1.3%). Infection with multiple species occurred in 9.4% of infections. After CD4 count was controlled for, rural residence (RR 1.40, 95% CI: 1.08-1.81), having no education (RR 1.57, 95% CI: 1.07-2.30), and higher CD4 count (RR 1.36, 95% CI: 1.07-1.73) remained independently associated with risk of helminth infection. Twelve weeks following treatment with albendazole, 32% of helminth-infected individuals had detectable helminths on examination. Residence, education, and CD4 count were not associated with persistent helminth infection. Among HIV-1 seropositive adults with CD4 counts above 250 cells/mm(3) in Kenya, traditional risk factors for helminth infection, including rural residence and lack of education, were associated with co-infection, while lower CD4 counts were not.


Radiation effective doses to patients undergoing abdominal CT examinations.
To determine the radiation effective dose to adult and pediatric patients undergoing abdominal computed tomographic (CT) examinations. Technique factors were obtained for three groups of randomly selected patients undergoing abdominal CT examinations: 31 children aged 10 years or younger; 32 young adults aged 11-18 years; and 36 adults older than 18 years. The radiographic techniques, together with the measured cross sections of patients, were used to estimate the total energy imparted to each patient. Each value of energy imparted was subsequently converted into the corresponding effective dose to the patient, taking into account the mass of the patient. All abdominal CT examinations were performed at 120 kVp with a section thickness of approximately 7 mm for all sizes of patients. The mean number of CT sections increased from 22.0 for children to 31.5 for adults, and the mean quantity of x radiation in milliampere-seconds increased from 220 mAs for children to 290 mAs for adults. The mean values (+/- SD) of energy imparted were 72.1 mJ +/- 24.4 for children, 183.5 mJ +/- 44.8 for young adults, and 234.7 mJ +/- 89.4 for adults. The corresponding mean values of patient effective dose were 6.1 mSv +/- 1.4 for children, 4.4 mSv +/- 1.0 for young adults, and 3.9 mSv +/- 1.1 for adults. Values of energy imparted to patients undergoing abdominal CT examinations were a factor of three higher in adults than in children, but the corresponding patient effective doses were 50% higher in children than in adults.


[The purity degree of the vagina containing mycoplasmosis urogenitalis (author's transl)].
In 156 women with mycoplasmosis urogenitalis the degree of purity of the vagina was determined. In none of the patient's a first degree of purity (containing lactobacilli only) was found. This is an additional sign that mycoplasma has an optional pathogenetic importance in the female urogenital system. In order to restore a normal vaginal flora of lactobacilli, the mycoplasma has to be eliminated.


Increased incidence of pathological and clinical prostate cancer with age: age related alterations of local immune surveillance.
The incidence of prostate cancer dramatically increases with age. The etiology still awaits elucidation, and the question as to why it is so prominently a disease of aging remains unanswered. We offer an explanation by suggesting an age related deficient immune surveillance. Reports published in the scientific literature with relevance to cancer and immunity, immune senescence, blood-prostate barrier and immune privilege were identified using MEDLINE. The existence of a blood-prostate barrier is a fair assumption, and the prostate may be considered an immune privileged site. With aging the prostate, a priori immunologically under surveilled, probably becomes more and more so. There is impaired function, transmigration and probably penetration into the gland of natural killer and other immune competent cells due to the onset of immune senescence coupled with impaired diapedesis and possible age related alterations in the blood-prostate barrier. Apparently, with aging the immune surveillance of the already immune privileged prostate is progressively and further affected. This condition may result in the inability of the gland to eradicate emergent malignant cells.


[Scintigraphic detection of pheochromocytoma metastases. Morphologic control apropos of a case].
The authors report a case of malignant pheochromocytoma. They explain the difficulties of the diagnostic of malignancy which can only be confirmed by metastases. Radioisotope scanning with I131 metaiodobenzyl guanidine is the best available method to locate the primary tumor and the metastases. The results of radio isotope scanning are compared to the ultrastructural study.


Formation of dTDP-rhamnose is essential for growth of mycobacteria.
It was determined that the dTDP-rhamnose synthesis gene, rmlD, could be inactivated in Mycobacterium smegmatis only in the presence of a rescue plasmid carrying functional rmlD. Hence, dTDP-rhamnose biosynthesis is essential for the growth of mycobacteria and the targeting of dTDP-rhamnose synthesis for new tuberculosis drugs is supported.


Dihydroxyadenine stone with adenine phosphoribosyltransferase deficiency: A case report.
Adenine phosphoribosyltransferase (APRT) deficiency is a rare autosomal recessive error of purine metabolism resulting in the generation of 2,8-dihydroxyadenine (DHA), a highly insoluble metabolite of adenine, which can cause radiolucent urolithiasis. This is the second case of DHA stone being reported in India and the first case in India to document the mutation of the APRT gene on blood DNA analysis.


Impact of Nutrient Restriction on the Structure of Listeria monocytogenes Biofilm Grown in a Microfluidic System.
Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30°C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA.


Fungal central nervous system infections: prevalence and diagnosis.
Fungal infections of the central nervous system (CNS) are rare but they pose a significant challenge. Their prevalence spans a wide array of hosts including immunosuppressed and immunocompetent individuals, patients undergoing neurosurgical procedures and those carrying implantable CNS devices. Cryptococcus neoformans and Aspergillus spp. remain the most common pathogens. Magnetic resonance imaging can help localize the lesions, but diagnosis is challenging since invasive procedures may be needed for the retrieval of tissue, especially in cases of fungal abscesses. Antigen and antibody tests are available and approved for use in the cerebrospinal fluid (CSF). PCR-based techniques are promising but they are not validated for use in the CSF. This review provides an overview on the differential diagnosis of the fungal CNS disease based on the host and the clinical syndrome and suggests the optimal use of diagnostic techniques. It also summarizes the emergence of Cryptococcus gatti and an unanticipated outbreak caused by Exserohilum rostratum.


Hemostatic resuscitation in traumatic hemorrhagic shock: case report.
The aim of this paper is to report a case in which the damage control resuscitation (DCR) approach was successfully used to promote hemostatic resuscitation in a polytraumatized patient with severe hemorrhagic shock. Female patient, 32 years of age, with severe hemorrhagic shock due to polytrauma with hip fracture, who developed acidosis, coagulopathy, and hypothermia. During fluid resuscitation, the patient received blood products transfusion of fresh frozen plasma/packed red blood cells/platelet concentrate at a ratio of 1:1:1 and evolved intraoperatively with improvement in perfusion parameters without requiring vasoactive drugs. At the end of the operation, the patient was taken to the intensive care unit and discharged on the seventh postoperative day. The ideal management of traumatic hemorrhagic shock is not yet established, but the rapid control of bleeding and perfusion recovery and well-defined therapeutic protocols are fundamental to prevent progression of coagulopathy and refractory shock.


Endoarterial scoring--a novel treatment for resistant pulmonary arterial lesions associated with Williams-Beuren syndrome.
A 7-year-old girl with Williams-Beuren syndrome had undergone multiple percutaneous interventions for resistant peripheral pulmonary arterial stenoses with primarily conventional high-pressure balloon dilatation, but also with use of a cutting balloon. On this occasion, we used the AngioSculpt (AngioScore, Alameda, California) to remodel her distal vessels bilaterally with good effect. We believe this to be the first report of its use in the pulmonary arterial circulation of man.


Acquired and intrinsic gemcitabine resistance in pancreatic cancer therapy: Environmental factors, molecular profile and drug/nanotherapeutic approaches.
A high number of cancer patients around the world rely on gemcitabine (GEM) for chemotherapy. During local metastasis of cancers, surgery is beneficial for therapy, but dissemination in distant organs leads to using chemotherapy alone or in combination with surgery to prevent cancer recurrence. Therapy failure can be observed as a result of GEM resistance, threatening life of pancreatic cancer (PC) patients. The mortality and morbidity of PC in contrast to other tumors are increasing. GEM chemotherapy is widely utilized for PC suppression, but resistance has encountered its therapeutic impacts. The purpose of current review is to bring a broad concept about role of biological mechanisms and pathways in the development of GEM resistance in PC and then, therapeutic strategies based on using drugs or nanostructures for overcoming chemoresistance. Dysregulation of the epigenetic factors especially non-coding RNA transcripts can cause development of GEM resistance in PC and miRNA transfection or using genetic tools such as siRNA for modulating expression level of these factors for changing GEM resistance are suggested. The overexpression of anti-apoptotic proteins and survival genes can contribute to GEM resistance in PC. Moreover, supportive autophagy inhibits apoptosis and stimulates GEM resistance in PC cells. Increase in metabolism, glycolysis induction and epithelial-mesenchymal transition (EMT) stimulation are considered as other factors participating in GEM resistance in PC. Drugs can suppress tumorigenesis in PC and inhibit survival factors and pathways in increasing GEM sensitivity in PC. More importantly, nanoparticles can increase pharmacokinetic profile of GEM and promote its blood circulation and accumulation in cancer site. Nanoparticles mediate delivery of GEM with genes and drugs to suppress tumorigenesis in PC and increase drug sensitivity. The basic research displays significant connection among dysregulated pathways and GEM resistance, but the lack of clinical application is a drawback that can be responded in future.


Construction of Fluorinated Benzoxathiin Skeleton by Successive Perfluorophenylthiolation/Cyclization of Activated α-Methylene Ketones by Perfluorophenyl Diethylaminosulfur Difluoride.
Pharmaceutically attractive fluorinated benzoxathiin (thiaflavan) skeleton 4 was directly constructed by the reaction of activated α-methylene ketones 1 such as β-keto esters, 1,3-diketone, and β-keto sulfones with a perfluorophenyl analogue of diethylaminosulfur trifluoride, C6F5-DAST, in high yields via successive perfluorophenylthiolation/cyclization reaction.


Endovascular treatment of anterior communicating artery aneurysms: results of clinical and radiological outcome in Newcastle.
The results of endovascular management of anterior communicating artery aneurysms (ACoAA) using Guglielmi Detachable Coils (GDC) are presented. We detail the clinical and radiological features, and postoperative clinical and radiological outcome in a consecutive series of patients. We have prospectively collected comprehensive data on our patients with SAH since 1989. This study reports on patients admitted between January 1990 and December 1998, and focuses on 30 patients who had their ACoAA endovascularly treated. An independent observer (TE) carried out long-term follow-up in January 2002. Statistical analysis was performed to study the relationship between clinical factors, the radiological morphology of aneurysms and the long-term outcome. The age ranged from 25 to 74 years (median: 54) and endovascularly treated ACoAA were more common in women, 19 (63%) compared with men, 11 (37%). Seventy-seven per cent were in good grade (WFNS 1 & 2) before treatment. Three patients rebled before treatment. The aneurysms were less than 10 mm in maximal diameter in 27/30 patients. The follow-up period was from 1 to 53 months (mean 32.5, median 36.6 months). Excellent outcome was recorded for 11 patients (36.7%), good in seven patients (23.3%), fair in three patients (10%) and poor in four cases (13.3%). Five patients had died (mortality 16.7%). Favourable outcome was achieved in younger patients, women, and in those who presented in grades 1 and 2. The long-term radiological follow-up revealed residual necks in 13 patients. Only one has required retreatment and no rebleed has occurred in 3-6 years. This study reports a contemporary experience with the endovascular management of ACoAA. Long-term stability of the coil and good outcome is demonstrated. Endovascular treatment using GDC is an efficient technique for treating acutely ruptured ACoAA with little additional morbidity. The clinical and radiological results are comparable with those in the literature.