Effects of dietary supplement on functional condition and clinical outcome measured with a modified Norton scale.
Elderly long-term care patients (n = 501) were randomly allocated to receive dietary supplementation (400 kcal/day) in addition to a normal hospital diet, or normal hospital diet only, for 26 weeks. The patients' functional condition was assessed by a modified Norton scale comprising mental condition, activity, mobility, food intake, fluid intake, incontinence and general physical state. Nutritional status was assessed by anthropometry, serum protein analysis and delayed hypersensitivity skin tests. Improvements during the first 8 weeks were seen in activity and general physical condition in the supplemented group, and in mobility and general physical condition in the control group. After 8 weeks the supplemented group showed a significantly higher level of activity (p < 0.05) compared to the control group. We conclude that in long-term geriatric care nutritional supplements help to maintain and improve the patients function.


Clinical and electrophysiological assessment of injury to infrapatellar branch(es) of saphenous nerve during anterior cruciate ligament reconstruction using oblique incision for hamstring graft harvest: A prospective study.
One of the complications of arthroscopic reconstruction of anterior cruciate ligament (ACL) using hamstring autograft is injury to the infrapatellar branch of the saphenous nerve (IPBSN) leading to loss of sensation around the knee. Its incidence varies according to the orientation of incision for hamstring graft harvest, with oblique incision having the lowest chance of this injury. Aim of this study was to assess clinically and electrophysiologically injury to IPBSN after ACL reconstruction using oblique incision for hamstring graft harvest. Fifty patients that were scheduled to undergo arthroscopic ACL reconstruction were included. All the patients were evaluated both clinically and electrophysiologically preoperatively and at three weeks and six months postoperatively to determine injury to IPBSN. Patient satisfaction after surgery was also compared. IPBSN injury was found to be present in 12 out of 50 patients (24%). Both clinical and electrophysiological data correlated completely. The difference in the length of the skin incisions between IPBSN injured and IPBSN intact patients was found to be statistically significant (P = .0043). The difference in the satisfaction score between patients with injured and intact IPBSN was also found to be statistically significant (P = .02). Oblique incision for hamstring graft harvest results in lower incidence of IPBSN injury and subsequent sensory loss when compared with similar studies with different skin incisions carried out in the past. Patients undergoing ACL reconstruction should be counseled preoperatively regarding this potential complication, and if this complication occurs, improvement can be expected with time.


Clinical evaluation of autoantibodies to liver cell membrane specific antigen, liver specific lipoprotein, and Tamm-Horsfall glycoprotein in autoimmune chronic active hepatitis.
An enzyme-linked immunosorbent assay was developed to detect circulating autoantibodies to three liver cell membrane surface antigens, i.e., liver cell membrane specific antigen (LCM), liver specific lipoprotein (LSP), and Tamm-Horsfall glycoprotein (THGP). In autoimmune chronic active hepatitis (autoimmune CAH), the positive rate and mean titer (normal range, less than 5.5 units) for anti-LCM were 100% and 13.5 units before corticosteroid treatment and 100% and 9.9 units during the treatment. The corresponding values for anti-LSP were 84% and 11.8 units, and 81% and 8.9 units, and those for anti-THGP were 84% and 12.3 units, and 81% and 7.9 units. In an autoimmune CAH patient, elevation of the plasma levels of autoantibodies during the treatment apparently preceded the elevation of alanine aminotransferase (ALT). However, the ALT elevation induced by transcatheter arterial embolization was not associated with the elevation of these autoantibodies in an autoimmune CAH patient with hepatocellular carcinoma. In primary biliary cirrhosis, drug-induced hepatitis, and non-hepatic immunological disorders, the production of the three autoantibodies did not directly correlate with liver cell damage. These findings suggest that the elevation of autoantibodies against LCM, LSP, and THGP can be a useful guide for the prednisolone treatment of autoimmune CAH.


Multiplexed detection and isolation of viable low-frequency cytokine-secreting human B cells using cytokine secretion assay and flow cytometry (CSA-Flow).
The ability to functionally characterize cytokine-secreting immune cells has broad implications in both health and a range of immune-mediated and auto-immune diseases. Low-frequency cytokine-defined immune-cell subsets can play key immune-regulatory roles, yet their detailed study is often hampered by limited clinical sample availability. Commonly used techniques including intracellular cytokine staining require cell fixation, precluding subsequent functional interrogation. The cytokine-secretion assay (CSA) can overcome this limitation, though has mostly been used for detection of relatively high-frequency, single-cytokine secreting cells. We examined how adaptation of the CSA in combination with multiparametric flow-cytometry (CSA-Flow) may enable simultaneous isolation of multiple, low-frequency, cytokine-secreting cells. Focusing on human B cells (traditionally recognized as harder to assay than T cells), we show that single-capture CSA-Flow allows for isolation of highly-purified populations of both low-frequency (IL-10+; GM-CSF+) and high-frequency (TNF+) cytokine-defined B cells. Simultaneous detection and isolation of up to three viable and highly-purified cytokine-secreting B-cell subpopulations is feasible, albeit with some signal loss, with fractions subsequently amenable to gene expression analysis and in vitro cell culture. This multiplexing CSA-Flow approach will be of interest in many human cellular immunology contexts aiming to functionally characterize cytokine-secreting immune cells, especially when sample volumes and cell numbers are limited.


Attachment across the lifespan: Examining the intersection of pair bonding neurobiology and healthy aging.
Increasing evidence suggests that intact social bonds are protective against age-related morbidity, while bond disruption and social isolation increase the risk for multiple age-related diseases. Social attachments, the enduring, selective bonds formed between individuals, are thus essential to human health. Socially monogamous species like the prairie vole (M. ochrogaster) form long-term pair bonds, allowing us to investigate the mechanisms underlying attachment and the poorly understood connection between social bonds and health. In this review, we explore several potential areas of focus emerging from data in humans and other species associating attachment and healthy aging, and evidence from prairie voles that may clarify this link. We examine gaps in our understanding of social cognition and pair bond behavior. Finally, we discuss physiologic pathways related to pair bonding that promote resilience to the processes of aging and age-related disease. Advances in the development of molecular genetic tools in monogamous species will allow us to bridge the mechanistic gaps presented and identify conserved research and therapeutic targets relevant to human health and aging.


Altitudinal variation in bumble bee (Bombus) critical thermal limits.
Organism critical thermal limits are often tightly linked to current geographic distribution and can therefore help predict future range shifts driven by changing environmental temperatures. Thermal tolerance of diverse organisms often varies predictably with latitude, with upper thermal limits changing little and lower thermal limits decreasing with latitude. Despite similarly steep gradients in environmental temperatures across altitude, few studies have investigated altitudinal variation in critical thermal limits. We estimated critical thermal minimum (CTmin), critical thermal maximum (CTmax) and recovery temperature (Trec) by tracking righting response of three bumble bee species during thermal ramps: Bombus huntii collected from 2180m asl, and Bombus bifarius and Bombus sylvicola collected from 3290m asl in Wyoming, USA. Overall, larger bees could tolerate more extreme temperatures, likely due to a thermal inertia driven lag between core body temperatures and air temperatures. Despite their smaller size, high altitude bumble bees tolerated colder air temperatures: they had ~1°C lower CTmin and recovered from cold exposure at ~3-4°C lower air temperatures. Conversely, low altitude bees tolerated ~5°C hotter air temperatures. These altitudinal differences in thermal tolerance parallel differences in average daily minimum (1.2°C) and maximum (7.5°C) temperatures between these sites. These results provide one of the few measurements of organism thermal tolerance across altitude and the first evidence for geographical differences in tolerance of temperature extremes in heterothermic bumble bees.


Fluorescence sensor array for identification of commercial milk samples according to their thermal treatments.
Identification of processed milk is of importance for commercial and legal concerns. The fluorescence response patterns induced by fluorophore/protein interactions allow a possible discrimination of processed milk samples corresponding to their thermal treatment. The fluorescence responses of 4 fluorophores upon addition of commercial milk samples in 96-well plate are measured in the range of 400-600 nm using the excitation wavelength at 375 nm. The pattern recognition of the 53,126 fluorescence responses (4 fluorophores×41 wavelengths×4 thermally processed milks×3 brands×3 lots×3 bottles×3 repeats) are analyzed by multivariate statistical methods. Linear discriminant analysis (LDA) successfully recognizes the milk samples according to their thermal processing, i.e. pasteurized milk, sterilized milk, UHT fresh milk and recombined milk (UHT milk having milk powder), with 100% classification accuracy in a cross validation using a leave-one-out technique.


"It was great to break down the walls between patient and provider": liminality in a co-produced advisory course for psychiatry residents.
Although rhetoric abounds about the importance of patient-, person- and relationship-centered approaches to health care, little is known about how to address the problem of dehumanization through medical and health professions education. One promising but under-theorized strategy is to co-produce education in collaboration with health service users. To this end, we co-produced a longitudinal course in psychiatry that paired people with lived experience of mental health challenges as advisors to fourth-year psychiatry residents at the University of Toronto. The goal of this study was to examine this novel, relationship-based course in order to understand co-produced health professions education more broadly. Using qualitative interviews with residents and advisors after the first iteration of the course, we explored how participants made meaning of the course and of what learning, if any, occurred, for whom and how. We found that the anthropological theory of liminality allowed us to understand participants' complex experiences and illuminated how this type of pedagogy may work to achieve its effects. Liminality also helped us understand why some participants resisted the course, and how we could more carefully think about co-produced, humanistic education and transformative learning.


Down-regulation of PvSAMS impairs S-adenosyl-L-methionine and lignin biosynthesis, and improves cell wall digestibility in switchgrass.
S-adenosyl- l-methionine (SAM) is the methyl donor involved in the biosynthesis of guaiacyl (G) and syringyl (S) lignins in vascular plants. SAM is synthesized from methionine through the catalysis of the enzyme S-adenosylmethionine synthase (SAMS). However, the detailed function of SAMS in lignin biosynthesis has not been widely investigated in plants, particularly in monocot species. In this study, we identified PvSAMS genes from switchgrass (Panicum virgatum L.), an important dual-purpose fodder and biofuel crop, and generated numerous transgenic switchgrass lines through PvSAMS RNA interference technology. Down-regulation of PvSAMS reduced the contents of SAM, G-lignins, and S-lignins in the transgenic switchgrass. The methionine and glucoside derivatives of caffeoyl alcohol were found to accumulate in the transgenic plants. Moreover, down-regulation of PvSAMS in switchgrass resulted in brownish stems associated with reduced lignin content and improved cell wall digestibility. Furthermore, transcriptomic analysis revealed that most sulfur deficiency-responsive genes were differentially expressed in the transgenic switchgrass, leading to a significant increase in total sulfur content; thus implying an important role of SAMS in the methionine cycle, lignin biosynthesis, and sulfur assimilation. Taken together, our results suggest that SAMS is a valuable target in lignin manipulation, and that manipulation of PvSAMS can simultaneously regulate the biosynthesis of SAM and methylated monolignols in switchgrass.


Nucleus anomaly test in Chinese hamster and in rat after treatment with isoniazid.
Nucleus anomaly test in Chinese hamsters and in rats treated with isoniazid (INH) was carried out according to a standard protocol in two different laboratories. These comprised both short-term studies, in which the tests were performed on animals killed 6, 12, 24, 36, or 48 h after the second of two consecutive doses of 5, 25, or 125 mg/kg INH given at an interval of 24 h, and long-term studies in animals treated with 25 mg/kg INH thrice weekly for 12 weeks. As a rule, each group consisted of at least four animals, and 1000 cells from each animal were examined. In one of the laboratories, a slight, but statistically significant increase in the incidence of nuclear anomalies was observed in two experiments on animals sacrificed 24 h after treatment; in the majority of cases, however, the investigations yielded negative results. Two out of three long-term studies revealed a slight, but statistically significant increase in the incidence of nuclear anomalies.


The effect of current and lifetime alcohol consumption on overall and central obesity.
To evaluate the association between current and lifetime alcohol consumption, and overall and central obesity in adults of an urban Portuguese population. Participants were randomly selected from the non-institutionalized Porto inhabitants, aged ≥18 years (EPIPorto Study: 1999-2003). The study included 2366 participants for evaluating current alcohol consumption and overall obesity, and 2377 participants for central obesity. A trained interviewer applied a questionnaire and anthropometrics were measured. Data on alcohol intake were obtained by using a food frequency questionnaire. Overall obesity was considered when body mass index was ≥30.0 kg/m(2), and central obesity when waist circumference (WC) was ≥88 cm in women and ≥102 cm in men. Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated, separately by sex, using unconditional logistic regression models. After adjustment for age, education, smoking, energy intake and regular physical exercise, men who consumed >60g/day of alcohol were more frequently obese compared with non-drinkers (OR=2.26, 95% CI: 1.17-4.35). When considering lifetime alcohol consumption, the magnitude of the association was stronger in both women (OR=2.41, 95% CI: 1.28-4.56) and men (OR=4.22, 95% CI: 1.93-9.24). For central obesity, women consuming 15.1-30 g/day and >30 g/day had higher probability of being obese, considering both current and lifetime alcohol consumption. In men, the same positive associations were observed, which were particularly stronger between lifetime alcohol consumption and central obesity (>60g/day vs non-drinkers: OR=4.02, 95% CI: 1.80-8.98). Independently of social and behavioural features, current and lifetime alcohol consumption were positively associated with overall and central obesity, in both women and men.


Interaction of Escherichia coli RNA polymerase with DNA in an elongation complex arrested at a specific psoralen crosslink site.
We have probed the interaction of Escherichia coli RNA polymerase with DNA in an elongation complex arrested by a site-specifically placed psoralen crosslink using DNase I footprinting techniques. The psoralen derivative 4'-hydroxymethyl-4,5',8-trimethylpsoralen was first placed at a specific site in the middle of a chemically synthesized double-stranded DNA fragment containing an E. coli RNA polymerase promoter at one end. The psoralen molecule was photochemically attached to two adjacent thymidine residues on opposite strands as a diadduct. Using this crosslinked DNA as the template for transcription, we found that the E. coli RNA polymerase was blocked at the psoralen diadduct, yielding a transcript 29 nucleotides long. The arrested elongation complex inhibited DNase I digestion of both the coding strand and the non-coding strand from about 22 nucleotides upstream to 15 nucleotides downstream from the diadduct. These results, which suggest that the unwindase and the catalytic sites of the polymerase are very close to each other, have been incorporated into a model of the transcription elongation complex.


The influence of psychological factors on tinnitus severity.
Subjective tinnitus is a frequent symptom characterized by perception of sound in the absence of a corresponding external stimulus. Although many people learn to live with tinnitus, some find it severely debilitating. Why tinnitus is debilitating in some patients, but not in others, is still incompletely understood. We aimed to assess the influence of different aspects of psychological distress on perceived tinnitus severity. Three hundred seventeen patients diagnosed with chronic subjective tinnitus at two university clinics completed the Tinnitus Handicap Inventory (THI), the Tinnitus Questionnaire (TQ) and the Symptom Check List-90-Revised. The influence of the different dimensions of psychological distress on perceived tinnitus severity was statistically evaluated. Both THI and TQ scores were significantly influenced by gender, site and the dimension "depression". In addition, TQ scores were significantly influenced by age and "somatization," whereas "hostility" had an impact on THI scores only. Psychological aspects as well as sociodemographic variables had a significant influence on both TQ scores. However, our results indicate, that these scales reflect emotional distress of tinnitus sufferers differently. This should be taken into consideration in the use of these scales as screening tools for assessment of tinnitus handicap.


What's new in diagnosis and treatment of HPV-associated cervical lesions.
The human papilloma virus (HPV) has been subject to intense discussion during the last decade, as evidence has accumulated to strongly suggest that viruses play an essential part in the genesis of cervical neoplasms. The contamination rate of the female population is reported to range between 5 and 80%, depending on the modality of detection. Many indications almost reliably suggest the oncogenicity of certain HPV-types. The highest progression rate was found in HPV 16 lesions, followed by HPV 18 lesions. In our material the incidence of HPV 16/18 and 31/33 infections compared to 6/11 was elevated in patients with cervical intraepithelial neoplasia (CIN) compared to women with non-CIN lesions. The exact histologic diagnosis of CIN lesions is essential for a differentiated therapy. Our data indicate that mild and moderate CIN lesions are detected more frequently by Pap smear and colposcopically directed punch biopsy than by cervical smears alone. Histological diagnosis and HPV-typing by in-situ hybridisation offers the possibility to establish a differentiated therapy, e.g. by way of close follow-up, local destruction or cone biopsy. Main attention should be directed at early detection of dysplasia, at timely institution of follow-up programs and a possibly conservative therapeutic management.


Radioisotopic labelling of endoalveolar surface-active phospholipidic fractions from rabbit pulmonary lavage.
A simple method of detecting endoalveolar tensioactive system fractions employing intravenously injected 131I-triolein has been devised. The tracer has been administered to 7 adult rabbit groups then sacrificed at different time intervals after the injection. Endoalveolar tensioactive system lipidic fractions collected by pulmonary lavage have been separated with thin-layer chromatography and their radioactivity evaluated by scanning plates. Endoalveolar tensioactive system fractions containing considerable amounts of C 18 fatty acids have been selectively labelled.


A male with two different familial autosomal fragile sites and a cytogenetically abnormal offspring.
During the course of routine prenatal cytogenetics, a male with 2 different autosomal fragile sites (FS) was detected. The FS were at 9p21 and 12q13 and his sister also had both fragile sites, inherited from an obligate carrier father. He was the father of a foetus with an abnormal chromosome complement 46,XY/47,XY,+frag. The origin of the fragment could not be determined.


Progression of deoxynivalenol and zearalenone concentrations in straw of wheat infected artificially withFusarium culmorum.
This investigation aimed at the progression of the contamination of theFusarium toxins deoxynivalenol (DON) and zearalenone (ZON) within the fractions straw, glumes and spindles from non-inoculated andFusarium-inoculated wheat. TheFusarium head blight (FHB)-susceptible wheat cultivar Ritmo was cultivated after the pre-crop maize and artificially infected withFusarium culmorum. Samples of whole wheat plants were taken once a week from anthesis until harvest and fractionated into straw, glumes and spindles. Samples were examined for deoxynivalenol and zearalenone and quantitatively determined by high performance liquid chromatography (HPLC) with diode-array detection (DAD) and fluorescence detection, respectively. Additionally, the impact of theFusarium inoculation on the crude protein content was scrutinised.Differences in the formation of deoxynivalenol and zearalenone with respect to date and concentration are shown by this trial. Deoxynivalenol was produced in higher concentrations and at earlier stages, whereas zearalenone was formed later and in smaller amounts. Furthermore, a rise of the deoxynivalenol concentration up to a maximum during the growing season, followed by a sudden decline at later stages until harvest, was observed. ThisFusarium infection resulted in an increased crude protein content in all of the three fractions.


Distinct Polymorphisms in HLA Class I Molecules Govern Their Susceptibility to Peptide Editing by TAPBPR.
Understanding how peptide selection is controlled on different major histocompatibility complex class I (MHC I) molecules is pivotal for determining how variations in these proteins influence our predisposition to infectious diseases, cancer, and autoinflammatory conditions. Although the intracellular chaperone TAPBPR edits MHC I peptides, it is unclear which allotypes are subjected to TAPBPR-mediated peptide editing. Here, we examine the ability of 97 different human leukocyte antigen (HLA) class I allotypes to interact with TAPBPR. We reveal a striking preference of TAPBPR for HLA-A, particularly for supertypes A2 and A24, over HLA-B and -C molecules. We demonstrate that the increased propensity of these HLA-A molecules to undergo TAPBPR-mediated peptide editing is determined by molecular features of the HLA-A F pocket, specifically residues H114 and Y116. This work reveals that specific polymorphisms in MHC I strongly influence their susceptibility to chaperone-mediated peptide editing, which may play a significant role in disease predisposition.


Descriptive assessment on diabetic retinopathy screening in an awareness programme in Malaysia.
To determine the prevalence of diabetic retinopathy (DR) among diabetic patients at the primary health clinics in Selangor, Malaysia. All diabetic patients were screened in Retinal Disease Awareness Programme (RDAP) and those who had significant DR changes were referred to the hospital for further management. Descriptive analyses were done to determine the prevalence of DR and sociodemographic characteristics among patients with diabetic. Univariate and multivariable analysis using Logistic regression were performed to find association and predictor factors in this screening. A total of 3305 patients aged 40y and above were screened for DR. Of the patients screened, 9% patients were found to have DR and other visual complication such as maculopathy (0.9%), cataract (4.8%) and glaucoma (0.4%). The mean age of patients without retinopathy was 57.82±8.470y and the mean age of patients with DR was 63.93±9.857y. About 61.5% of the patients screened were aged below 60y and 38.5% were aged 60y and above. Majority of the patients screened were women 58.5% and Malay in the age group of 50-59y, while 27% were aged 60-69y. Significant association were found between age, sex, race, visual loss and DR. Although the prevalence of DR among patients is not alarming, effective interventions need to be implemented soon to avert a large burden of visual loss from DR.


Responses to digital disinformation as part of hybrid threats: a systematic review on the effects of disinformation and the effectiveness of fact-checking/debunking.
The dissemination of purposely deceitful or misleading content to target audiences for political aims or economic purposes constitutes a threat to democratic societies and institutions, and is being increasingly recognized as a major security threat, particularly after evidence and allegations of hostile foreign interference in several countries surfaced in the last five years. Disinformation can also be part of hybrid threat activities. This research paper examines findings on the effects of disinformation and addresses the question of how effective counterstrategies against digital disinformation are, with the aim of assessing the impact of responses such as the exposure and disproval of disinformation content and conspiracy theories. The paper's objective is to synthetize the main scientific findings on disinformation effects and on the effectiveness of debunking, inoculation, and forewarning strategies against digital disinformation. A mixed methodology is used, combining qualitative interpretive analysis and structured technique for evaluating scientific literature such as a systematic literature review (SLR), following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) framework.


Cholinergic mechanisms in cocaine-induced genital reflexes in paradoxical sleep-deprived male rats.
In view of the fact that paradoxical sleep deprivation (PSD) modifies cocaine-induced genital reflexes (penile erection [PE] and ejaculation [EJ]), the aim of this study was to address the interaction of cholinergic agents with the action of cocaine on the genital reflexes of PSD male rats. After a 4-day period of PSD, each group was administered with cholinergic drugs 1 h prior to cocaine and was placed in observation cages. The administration of nicotine (0.12, 0.25, 0.5 and 1 mg/kg sc) reduced the frequency and number of animals displaying PE and increased PE latency. Pretreatment with mecamylamine (1.25, 5, 10 and 20 mg/kg sc) also significantly reduced PE frequency for all doses used. The percentage of rats showing EJ was significantly reduced in the group pretreated with 1 mg/kg of nicotine compared with the saline group. The administration of pilocarpine (1.25, 2.5, 5 and 10 mg/kg sc) and atropine (1.25, 5, 10 and 20 mg/kg sc) led to a reduction in the frequency of PE displayed by the rats. These data show that agonist and antagonist cholinergic drugs inhibit genital reflexes in PSD male rats injected with cocaine. The data also suggest that the stimulating action of cocaine in potentiating the sexual effects in PSD rats does not override the effects of the cholinergic mechanisms of sexual behavior.


Current Status on Extracellular Vesicles from the Dimorphic Pathogenic Species of Paracoccidioides.
Species of the Paracoccidioides brasiliensis complex and P. lutzii cause human paracoccidioidomycosis (PCM). Early interest in Paracoccidioides extracellular vesicles (EVs) resulted in a series of publications that unraveled the EVs protein, carbohydrate, lipid, and RNA cargo from isolates of different phylogenetic groups and distinct virulence degrees. EV and cell wall components were compared. These works are discussed in parallel with more recent data on the role of Paracoccidioides EVs in immunomodulation.


Imaging microstructural damage and plasticity in the hippocampus during epileptogenesis.
Epileptogenesis refers to the development and extension of tissue capable of generating spontaneous seizures, resulting in the development of an epileptic condition and/or progression of epilepsy after the condition is established. The hippocampus is the seizure-initiating zone in many epilepsy patients as well as in animal models of epilepsy. During epileptogenesis, the hippocampus undergoes structural changes, including mossy fiber sprouting; alterations in dendritic branching, spine density, and shape; and neurogenesis. In vivo magnetic resonance imaging (MRI) techniques provide insights into the microstructural organization of the hippocampus. An assessment of the structural plasticity of the hippocampus may provide parameters that could be used as biomarkers for epileptogenesis. Here we review conventional and more advanced MRI methods for detecting hippocampal tissue changes related to epileptogenesis. In addition, we summarize how diffusion tensor imaging can reveal cellular damage and plasticity, even at the level of hippocampal subfields. Finally, we discuss challenges and future directions for using novel MRI techniques in the search of biomarkers associated with epileptogenesis after brain injury.


Location of high and low affinity fatty acid binding sites on human serum albumin revealed by NMR drug-competition analysis.
Human serum albumin (HSA) is an abundant plasma protein that transports a wide variety of drugs and endogenous compounds. The complex binding capacity of HSA has made it a challenging system to study in detail but in order to develop our understanding of the interactions between ligands for HSA, the locations and relative affinities of different ligand binding sites must be determined. Albumin possesses multiple binding sites for its primary physiological ligand, non-esterified fatty acids (FA). Previously, titration of BSA with (13)C-labeled FA revealed multiple chemical shifts and allowed identification of a subset of three chemical shifts that were associated with high affinity FA binding. Recent crystallographic studies of HSA have mapped at least seven FA binding sites for long-chain FA and delineated the overlap with binding sites for drugs and other endogenous compounds. We aim to correlate NMR and structural data for FA to provide a more complete description of the binding capacity of HSA. Our recent mutagenesis studies allowed us to identify two high affinity binding sites in domain III of HSA. Here, we use NMR to study the binding of (13)C-carboxyl labeled palmitate to HSA in the presence and absence of competitor ligands to complete the correlation of NMR chemical shifts with specific structural binding sites. We carefully selected ligands with specific binding sites identified by crystallography and used them, either singly or in combination, to compete with [(13)C]palmitate for binding to HSA. We show that FA sites 2, 4 and 5 bind FA with high affinity, while sites 1, 3, 6 and 7 exhibit low affinity for FA, thus providing the first complete determination of relative affinities of all seven long-chain FA sites on HSA. Our results also yield direct insights into the interactions between FA and other ligands.


Stepwise differentiation from naïve state pluripotent stem cells to functional primordial germ cells through an epiblast-like state.
A group of pluripotent cells appearing during mammalian embryogenesis is the source for all the cell lineages that compose the embryo proper. In mice, pluripotent cells are first established in the inner cell mass (ICM) of the preimplantation blastocyst. After implantation, the ICM soon transforms into a cup-shaped epithelium, called the postimplantation epiblast. The two types of pluripotent cells, the ICM and postimplantation epiblast cells, are distinct, based on the differences in their gene expression profiles, epigenetic status, and differentiation capacity. During gastrulation, some of the postimplantation epiblast cells adjacent to the extraembryonic ectoderm are specified as primordial germ cells (PGCs), precursors of the germ cell lineage, in response to bone morphogenetic protein 4 (BMP4). Recently, we succeeded in reconstituting epiblast differentiation and PGC specification in vitro using pluripotent stem cells. Here, we describe the culture method of a stepwise differentiation from pluripotent stem cells to functional PGCs.


[How I explore … abdominal tuberculosis in children ?]
Tuberculosis (TB) is one of the top 10 causes of death worldwide. Abdominal TB is an uncommon presentation of TB and is challenging to diagnose due to its insidious onset. The most common forms in children are peritonitis and lymphadenitis. Diagnosis is based on radiological and histopathological findings. Specific PCR amplification confirms the diagnosis quicker than conventional cultures. The treatment includes a 6-month therapy and a close follow-up. This article describes the different methods allowing to confirm the diagnosis of abdominal TB.


Standardisation of 11C.
The increasing use of positron emission tomography for medical imaging and the availability of short-lived positron emitters has raised concerns about the accuracy of calibration of secondary standard measurement systems and the viability of using a single long-lived positron emitter as a reference calibration source for all positron emitters. Potential problems arise because the 511 keV quanta arising from positron annihilation are not generally produced at the same point as the original disintegration. In addition, the secondary standard may also be responsive to the associated bremsstrahlung radiation. The magnitude of both effects depends on the positron end-point energy. In order to resolve these problems, it is necessary to produce absolute standards of these positron-emitting radionuclides and the work presented here details the results of such work with 11C.


Ataxic Guillain-Barré syndrome associated with anti-GM1b and anti-GalNAc-GD1a antibodies.
Ataxic Guillain-Barré syndrome (GBS) associated with anti-GQ1b IgG antibody has been reported. We, however, have had a patient with ataxic GBS who had IgG antibodies to the minor gangliosides GM1b and GalNAc-GD1a, and we therefore retrospectively investigated the clinical features of patients who had antibodies to GM1b or GalNAc-GD1a, but not to GQ1b. Information on patients' antecedent illnesses, initial symptoms, neurological signs, and CSF findings was reviewed in those with ataxic GBS or Fisher syndrome (FS) with anti-GM1b or anti-GalNAc-GD1a IgG antibodies. We tested whether the anti-GM1b and anti-GalNAc-GD1a antibodies are cross-reactive and constructed three-dimensional structural models of GM1b and GalNAc-GD1a. Ataxic GBS was diagnosed in 1 of 65 patients who had both anti-GM1b and anti-GalNAc-GD1a antibodies and in 3 of 159 patients who had anti-GM1b antibody without anti-GalNAc-GD1a antibody: FS was diagnosed in 1 of the 159 patients and in 1 of 35 who had anti-GalNAc-GD1a antibody without anti-GM1b antibody. All the patients' antibodies to GM1b or GalNAc-GD1a were associated with the IgG isotype. The clinical features of patients with ataxic GBS associated with anti-GM1b or anti-GalNAc-GD1a IgG antibodies did not differ from those of patients who had anti-GQ1b IgG antibody. Absorption study findings for serum from the patient who had both anti-GM1b and anti-GalNAc-GD1a IgG antibodies showed significant absorbance of anti-GM1b IgG antibody by GalNAc-GD1a and of anti-GalNAc-GD1a IgG antibody by GM1b, indicating that these antibodies are cross-reactive. This is the first report of ataxic GBS or FS associated with anti-GM1b or anti-GalNAc-GD1a IgG antibodies. These autoantibodies, as well as anti-GQ1b IgG antibody, may function in the development of some patients with ataxic GBS and FS.


Allogeneic haematopoietic stem cell transplant in Philadelphia-positive acute lymphoblastic leukaemia.
ALL in which the Philadelphia (Ph) chromosome is detected is one of the few diseases in which there is almost unequivocal agreement that a matched sibling allogeneic haematopoietic stem cell transplant in first CR is the most appropriate therapy for patients within certain age limits. Extension of allogeneic stem cell transplant to patients without matched sibling donors or to older individuals is increasingly possible due to unrelated donors, umbilical cord blood and reduced-intensity conditioning regimens. Here, we carefully review evidence supporting current practice and examine recent evidence relating to the use of newer allogeneic transplant technologies in Ph-pos ALL. We explore the burgeoning literature on the role of tyrosine kinase inhibitors in this disease and summarize their impact on the transplant practice.


Flanged intraocular lens fixation via 27-gauge trocars using a double-needle technique decreases surgical wounds without losing its therapeutic effect.
Intraocular lens (IOL) fixation using a sutureless 27-gauge needle intrascleral IOL implantation technique requires six surgical wounds. We developed a modified technique using two 27-gauge trocars for vitrectomy and indwelling flanged IOL haptics to reduce the number of surgical wounds. Department of Ophthalmology, Hyogo College of Medicine. This retrospective study enrolled 54 patients who had undergone IOL scleral fixation between January 2016 and April 2019. Patients who underwent IOL scleral fixation and were observed for >12 weeks were analysed using medical record data. Before October 2017, patients underwent normal flanged IOL scleral fixation. Between November 2017 and April 2019, patients underwent the modified method (flanged IOL via 27-gauge trocars with double-needle technique). Primary end-point was superiority or non-inferiority of modified IOL scleral fixation compared with normal IOL scleral fixation for visual acuity (VA). Changes in corneal endothelium cell number, refractivity, astigmatisms and surgery-related complications, were secondary end-points. There were no significant differences in baseline characteristics between groups. Raw VA and best collected VA (BCVA) were significantly improved in all eyes (p-). There were no statistical significances in raw VA, BCVA and surgery-related complications between groups. Results of the modified technique were not inferior compared with the original technique although it was less invasive. Therefore, flanged IOL fixation via trocars using a double-needle technique is a useful technique for IOL implantation.


Retinoids induce stem cell differentiation via epigenetic changes.
Vitamin A (all-trans retinol) and its active metabolites, collectively called retinoids, exert potent effects on stem cell differentiation and thus, the formation of the entire organism, in part via the modulation of the epigenome. All-trans retinoic acid (RA), through binding to the retinoic acid receptors (RARs), alters interactions of the RARs with various protein components of the transcription complex at numerous genes in stem cells, and some of these protein components of the transcription complex then either place or remove epigenetic marks on histones or on DNA, altering chromatin structure and leading to an exit from the self-renewing, pluripotent stem cell state. Different epigenetic mechanisms, i.e. first, primarily H3K27me3 marks and then DNA methylation, may be employed by embryonic stem cells and other stem cells for control of early vs. late stages of cell differentiation. Creating these stable epigenetic changes requires the actions of many molecules, including tet1, polycomb protein complexes (PRCs), miRNAs, DNA methyltransferases (DNMTs), and telomerase reverse transcriptase. A more complete understanding of retinoid-dependent stem cell differentiation should reward us with new insights into the failure to maintain a differentiated state that is an essential part of neoplastic cell transformation and cancer.


Outcomes of radical debridement versus no debridement for the treatment of thoracic and lumbar spinal tuberculosis.
Whether radical debridement is necessary for the treatment of thoracic and lumbar tuberculosis is still questionable. The objective of this prospective randomized study was to compare the outcomes of radical debridement versus no debridement for the treatment of thoracic and lumbar tuberculosis. Seventy-four thoracic and lumbar tuberculosis patients with a neurological function of grade D and E underwent surgery and received the same chemotherapy regiment from January 2009 to October 2014. All patients were divided into group A and B by taking the drawing of lots. In group A, radical debridement, bone graft, and instrumentation were performed. Isolated posterior instrumentation without debridement were performed in group B. The operative time, blood loss, visual analogue score (VAS), erythrocyte sedimentation rate (ESR), kyphotic angle, Frankel grading, fusion rate, and complications were evaluated. Group B had a better clinical outcome with regard to the operative time, blood loss, VAS score first week post-operatively, and the ESR value in the third and sixth month post-operatively than group A, and the differences between the two groups about those values all presented a significant difference (P < 0.05). However, no difference was observed between the two groups for the kyphotic angle (P = 0.088) and fusion rate (P = 0.164) at the final follow-up. Neurological function of all cases exhibited normal neurological function in the two groups at the final follow-up. Two cases of pulmonary infection and four cases of wound infection in group A. No serious complications were observed in group B. Isolated posterior instrumentation without debridement is a suitable treatment for selected patients because of minor surgical trauma, fewer complications, and spontaneous fusion.


A Cannabinoid Receptor 2 Agonist Prevents Thrombin-Induced Blood-Brain Barrier Damage via the Inhibition of Microglial Activation and Matrix Metalloproteinase Expression in Rats.
Thrombin mediates the life-threatening cerebral edema and blood-brain barrier (BBB) damage that occurs after intracerebral hemorrhage (ICH). We previously found that the selective cannabinoid receptor 2 (CB2R) agonist JWH-133 reduced brain edema and neurological deficits following germinal matrix hemorrhage (GMH). We explored whether CB2R stimulation ameliorated thrombin-induced brain edema and BBB permeability as well as the possible molecular mechanism involved. A total of 144 Sprague-Dawley (S-D) rats received a thrombin (20 U) injection in the right basal ganglia. JWH-133 (1.5 mg/kg) or SR-144528 (3.0 mg/kg) and vehicle were intraperitoneally (i.p.) injected 1 h after surgery. Brain water content measurement, Evans blue (EB) extravasation, Western blot, and immunofluorescence were used to study the effects of a CB2R agonist 24 h after surgery. The results demonstrated that JWH-133 administration significantly decreased thrombin-induced brain edema and reduced the number of Iba-1-positive microglia. JWH-133 also decreased the number of P44/P42(+)/Iba-1(+) microglia, lowered Evans blue extravasation, and inhibited the elevated matrix metallopeptidase (MMP)-9 and matrix metallopeptidase (MMP)-12 activities. However, a selective CB2R antagonist (SR-144528) reversed these effects. We demonstrated that CB2R stimulation reduced thrombin-induced brain edema and alleviated BBB damage. We also found that matrix metalloproteinase suppression may be partially involved in these processes.


The mechanism of the warfarin-rifampin drug interaction in humans.
The mechanism of the drug interaction in humans between warfarin and rifampin was investigated by monitoring the elimination kinetics and metabolic disposition of a single oral dose of pseudoracemic warfarin by GC/MS. The decrease in hypoprothrombinemia observed with concomitant administration of therapeutic doses of rifampin was accompanied by a substantial decrease in the elimination half-lives of both warfarin enantiomers. Rifampin increased the clearance of (R)-warfarin threefold and the clearance of (S)-warfarin twofold. The excretion profiles for warfarin and its metabolites in urine and feces were similar for both control and treated subjects with the exception that 4'-hydroxywarfarin (stereoselective for the (S)-enantiomer) was observed when rifampin was administered. 4'-Hydroxywarfarin is a metabolite of the drug hitherto undetected in vivo in humans. Based on formation clearance values estimated for 6-, 7-, and 8-hydroxywarfarin, rifampin appears to increase the clearance of the parent drug by induction of the cytochrome P-450 isozyme(s) responsible for aromatic hydroxylation.


Do needs, symptoms or disability of outpatients with schizophrenia influence family burden?
Most needs of outpatients with schizophrenia are met by the family. This could cause high levels of family burden. The objective of this study is to assess the relationship between the patients' needs and other clinical and disability variables and the level of family burden. A total sample of 231 randomly selected outpatients with schizophrenia was evaluated with the Camberwell Assessment of Needs, Positive and Negative Syndrome Scale, Global Assessment of Functioning and Disability Assessment Scale. A total of 147 caregivers also answered the objective and subjective family burden questionnaire (ECFOS-II). Correlations between total number of needs and family burden, t tests between presence or absence of need for each domain of family burden and regression models between family burden and needs, symptoms, disability and sociodemographic variables were computed. The number of patients' needs was correlated with higher levels of family burden in daily life activities, disrupted behaviour and impact on caregiver's daily routine. The patients' needs most associated with family burden were daytime activities, drugs, benefits, self-care, alcohol, psychotic symptoms, money and looking after home. In a regression model, a higher number of needs, higher levels of psychopathology and disability, being male and older accounted for higher levels of family burden. Patients with schizophrenia with more needs cause greater family burden but not more subjective concerns in family members. The presence of patients' needs (daytime activities, alcohol and drug), severity of psychotic symptoms and disability are related to higher levels of family burden.


Efficacy of blood urea nitrogen and the neutrophil-to-lymphocyte ratio as predictors of mortality among elderly patients with genitourinary tract infections: A retrospective multicentre study.
To investigate whether initial blood urea nitrogen (BUN) and the neutrophil-to-lymphocyte ratio (NLR) in the emergency department (ED) are associated with mortality in elderly patients with genitourinary tract infections. A total of 541 patients with genitourinary tract infections in 5 EDs between November 2016 and February 2017 were included and retrospectively reviewed. We assessed age, sex, comorbidities, vital signs, and initial laboratory results, including BUN, NLR and the SOFA criteria. The primary outcome was all-cause in-hospital mortality. The nonsurvivor group included 32 (5.9%) elderly patients, and the mean arterial pressure (MAP), NLR and BUN were significantly higher in this group than in the survivor group (p < 0.001, p = 0.003, p < 0.001). In multivariate analysis, MAP <70 mmHg, NLR ≥23.8 and BUN >28 mg/dl were shown to be independent risk factors for in-hospital mortality (OR 3.62, OR 2.51, OR 2.76: p = 0.002, p = 0.033, p = 0.038, respectively). Additionally, NLR ≥23.8 and BUN >28 were shown to be independent risk factors for mortality in admitted elderly with complicated UTI (p = 0.030, p = 0.035). When BUN and NLR were combined with MAP, the area under the ROC curve (AUROC) value was 0.807 (0.771-0.839) for the prediction of mortality, the sensitivity was 87.5% (95% CI 71.0-96.5), and the specificity was 61.3% (95% CI 56.9-65.5%). The initial BUN and NLR values with the MAP were good predictors associated with all-cause in-hospital mortality among elderly genitourinary tract infections visiting the ED.


MyoD binds to the guanine tetrad nucleic acid structure.
A high affinity interaction between a protein and the guanine tetrad nucleic acid structure is described. Recombinant MyoD, a transcription factor that can initiate myogenesis, specifically bound to helical structures formed by stacks of guanine residues in square planar arrays. The N-7 methylation of a set of consecutive dG residues in a single-stranded probe of the creatine kinase enhancer interfered with the formation of this nucleic acid structure and prevented protein binding. Recombinant MyoD also bound to a guanine tetrad formed with a telomeric DNA probe, and it had a higher affinity for the four-stranded structure than for the double-stranded E-box-binding site. These data are the first report of a direct interaction between a protein and this nucleic acid conformation. The potential biological significance of this finding is discussed.


Farming-up coastal fish assemblages through a massive aquaculture escape event.
We investigated the changes on the mean trophic level of fish assemblages across different spatiotemporal scales, before and after a massive escape event occurred off La Palma (Canary Islands), which resulted in the release of 1.5 million fish (mostly Dicentrarchus labrax) into the wild. The presence of escaped fish altered significantly the mean trophic level of fish assemblages in shallow coastal waters. This alteration was exacerbated by the massive escape. A nearby marine protected area buffered the changes in mean trophic level but exhibited the same temporal patterns as highly fished areas. Moreover, escaped fish exploited natural resources according to their total length and possibly, time since escapement. New concerns arise as a "farming up" process is detected in shallow coastal fish assemblages where marine aquaculture is established.


Simplified Manouguian's aortic annular enlargement for aortic valve replacement.
A simplified Manouguian's aortic annular enlargement for aortic valve replacement with prosthetic valve was performed in a patient with severe aortic stenosis and mild regurgitation by preserving both the anterior mitral leaflet and the left atrial roof intact. This method minimized the operative risk by avoiding injury to the mitral valve, while inserting a possibly two-size-larger prosthetic valve. The inserted prosthetic valve was positioned parallel to the original aortic valve. Moreover, the incision from the top of the commissure between the noncoronary cusp and left coronary cusp through the interventricular fibrous trigone appears to be easily extended into the exact center of the anterior mitral leaflet if further enlargement is required.


Prediction model for unsuccessful return to work after hospital-based intervention in low back pain patients.
Many studies on low back pain (LBP) have identified prognostic factors, but prediction models for use in secondary health care are not available. The purpose of this cohort study, based on a randomised clinical study, was to identify risk factors for unsuccessful return to work (U-RTW) in sick-listed LBP patients with or without radiculopathy and to validate a prediction model for U-RTW. 325 sick-listed LBP patients with or without radiculopathy were included in an intervention study and followed for one year. Afterwards, 117 other LBP patients were recruited similarly, included in a validation study and also followed for one year. All patients were subjected to identical procedures and interventions and received a brief intervention by the same rehabilitation doctor and physiotherapist. Half of them received case manager guidance within a multidisciplinary setting. At baseline, they completed a questionnaire and went through a clinical low-back examination. Sciatica was investigated by magnetic resonance imaging (MRI). U-RTW was registered in a national database both initially and at 1-year. Neither initial U-RTW (24.0%) nor one-year U-RTW (38.2%) were statistically significantly different in the two intervention groups nor in patients with and without radiculopathy. Multivariate logistic regression analysis identified two clinical and five psychosocial baseline predictors for one-year U-RTW (primary outcome). The clinical predictors included pain score (back+leg pain) and side-flexion. The five psychosocial predictors included 'bodily distress' 'low expectations of RTW', 'blaming the work for pain', 'no home ownership' and 'drinking alcohol less than once/month'. These predictors were not statistically significantly different in patients with and without radiculopathy, and they also predicted initial U-RTW (secondary outcome). Obesity and older age were only supplementary predictors in patients with radiculopathy. A prediction model was established and tested in the validation study group. The model predicted one-year U-RWT in patients with intermediate and high risk, but only partially in patients with low risk. The model predicted all three risk categories in initial U-RTW. A prediction model combining baseline clinical and psychosocial risk factors predicted patients with low, intermediate and high risk for unsuccessful return to work, both initially and at 1-year.


Autonomic function in patients with functional dyspepsia assessed by 24-hour heart rate variability.
The function of the autonomic nervous system was assessed in 23 patients with dysmotility-like functional dyspepsia and 12 healthy volunteers by analysis of 24-hr heart rate variability and a battery of five standardized sympathetic and parasympathetic cardiovascular reflex tests. Measures of heart rate variability were obtained by analysis of ambulatory electrocardiographic recordings using both the time and the frequency domain methods. The values of parameters reflecting vagal activity (HF, rMSSD) were significantly lower in patients with functional dyspepsia. Individual analysis of heart rate variability data identified at least one abnormal parameter of vagal function in seven (30.4%) patients, and in five of these the results of parasympathetic cardiovascular reflex tests were also abnormal. Our results suggest impaired efferent vagal function in a subgroup of patients with functional dyspepsia. This abnormality may play a role in the pathogenesis of the disease in these patients.


Acetylation phenotyping of isoniazid using a simple and accurate high-performance liquid chromatography.
A simple, specific, accurate and reproducible method for the analysis of isoniazid and its major metabolite, N-acetylisoniazid in urine using high-performance liquid chromatography (HPLC) is described. The assay is performed after extraction of isoniazid, N-acetylisoniazid and 5-(4-methylphenyl)-5-phenylhydantoin (internal standard) from urine using a mixture of chloroform:isopropanol (70:30, v/v) and eluted from a 5 microns C-18 reversed phase column at ambient temperature with a mobile phase consisting of 10 mM sodium acetate:methanol:acetonitrile (40:40:20, v/v) containing 10 mM dioctylsulphosuccinate sodium and adjusted to pH 2.9 with sulphuric acid (less than 1 ml), at a flow rate of 1 ml/min with u.v. detection at 266 nm. Quantification was achieved by the measurement of the peak height ratio, and the absolute recoveries ranged from 94 to 99%. Within-day coefficients of variation ranged from 2.81 to 4.54% for isoniazid and from 2.37 to 3.75% for N-acetylisoniazid. Between-day CVs varied from 3.27 to 5.62% and from 2.5 to 4.91% for isoniazid and N-acetylisoniazid, respectively. Preliminary stability tests using a urine sample from a subject showed an increase in mean isoniazid concentration of about 25% after 1 month storage at -20 degrees C. The method was used for acetylation phenotyping of five individuals.


Regulation of EP4 expression via the Sp-1 transcription factor: inhibition of expression by anti-cancer agents.
For glioblastomas, COX-2 expression is linked to poor survival. COX-2 effects are mediated by the receptors EP2 and EP4, whose regulation is poorly understood. The expression of EP4, and activation or inhibition of EP4 activity in human glioblastoma T98G cells, was found to correlate with growth on soft agar. Chemoprevention drugs, troglitazone (TGZ) and some COX inhibitors, significantly suppressed EP4 expression in T98G cells in a dose dependant manner. Specificity protein 1 (Sp-1) binding sites, located within region -197 to -160 of the human EP4 promoter, are important for the transcription initiation of the human EP4 gene and are responsible for the EP4 suppression by TGZ. Mutation in the Sp-1 sites altered the promoter activity of luciferase constructs and TGZ effects on the promoter. The inhibitory effect of TGZ on EP4 expression was reversed by PD98059, a MEK-1/Erk inhibitor. Immunoprecipitation-Western blot analysis detected Sp-1 phosphorylation that was dependent on TGZ-induced Erks activation. ChIP assay confirmed that Sp-1 phosphorylation decreases its binding to DNA and as a result, leads to the suppression of EP4 expression. Thus, we propose that the expression of EP4 is regulated by Sp-1, but phosphorylation of Sp-1 induced by TGZ suppresses this expression. This represents a new and unique mechanism for the regulation of the EP4 receptor expression.


Oscillating CD8(+) T cell effector functions after antigen recognition in the liver.
When hepatitis B virus (HBV)-specific CD8(+) cytotoxic T lymphocytes (CTLs) are adoptively transferred into HBV transgenic mice, they enter the liver, recognize antigen, secrete interferon gamma (IFNgamma), inhibit viral replication, and kill their target cells, causing hepatitis. In the current study, we examined the impact of antigen recognition on the evolution of the activation phenotype, antiviral effector functions, expansion and contraction kinetics, and compartmentalization of the transferred CTLs. The results reveal that noncytolytic and cytolytic effector functions and expansion-contraction kinetics of the CTLs are regulated asynchronously and in an oscillatory manner as a consequence of antigen recognition in the liver and in association with PD-1 upregulation. We suggest that such oscillations maximize viral clearance and minimize tissue injury during HBV infection and that poor coordination of these events could lead to viral persistence and chronic liver disease.


Identification of multiple ral gene products in human platelets that account for some but not all of the platelet Gn-proteins.
Polyclonal antibodies raised against specific recombinant low molecular mass GTP-binding proteins were tested for their ability to recognize partially purified human platelet membrane Gn-proteins (i.e. proteins that bind [alpha-32P]GTP on nitrocellulose blots of SDS/polyacrylamide gels). An antiserum against simian ralA protein recognized a 27 kDa human platelet protein with the same apparent molecular mass as the major platelet Gn-protein (Gn27). In further analysis by two-dimensional polyacrylamide gel electrophoresis, the isoelectric focusing step permitted resolution of 12 major Gn-protein forms, seven of 27 kDa (Gn27a-g), one of 26 kDa (Gn26) and four of 24 kDa (Gn24a-d). The ralA antibody reacted strongly with the five most basic Gn27 species (a-e), weakly with Gn26 and not at all with Gn27f, Gn27g or Gn24a-d. We conclude that ral gene products account for some but probably not for all of the platelet Gn-proteins.


[Reconstruction of the aorta in mycotic aortic aneurysm--report of two cases of salmonella infection (author's transl)].
Two patients with infected aortic aneurysm underwent surgical treatment. When using autologous tissue in place of the infected aorta a recurrent aneurysm was observed four months later. Extraanatomical bypass of the infected area with prosthetic material and consecutive removal of all infected tissue seems to be the only successful management. This is confirmed by another patient who was treated successfully in this way. Prolonged antibiotic therapy after resection of infected aneurysms seems to be mandatory.


Benchmarking Cationic Monolayer Protected Nanoparticles and Micelles for Phosphate-Mediated and Nucleotide-Selective Proton Transfer Catalysis.
Both micelles and self-assembled monolayer (SAM)-protected nanoparticles are capable of efficiently hosting water-immiscible substrates to carry out organic reactions in aqueous media. Herein, we have analyzed the different catalytic effect of SAM-protected cationic nanoparticles and cationic surfactants of varying chain length towards base-catalyzed proton transfer mediated ring-opening reaction of 5-nitrobenzisoxazole (NBI) (also known as Kemp Elimination (KE) reaction). We use inorganic phosphate ion or different nucleotide (phosphate-ligated different nucleoside) as base to promote the reaction on micellar or nanoparticle interface. We find almost 2-3 orders of magnitude higher concentration of surfactants of comparable hydrophobicity required to reach the similar activity which attained by low cationic head group concentration bound on nanoparticle. Additionally, at low concentration of nanoparticle-bound surfactant or with high surfactant in micellar form, nucleotide-selectivity has been observed in activating KE reaction unlike free surfactant at low concentration. Finally, we showed enzyme-mediated nucleotide hydrolysis to generate phosphate ion which in situ upregulate the KE activity much more in GNP-based system compared to CTAB. Notably, we show a reasonable superiority of SAM-protected nanoparticles in activating chemical reaction in micromolar concentration of headgroup which certainly boost up application of SAM-based nanoparticles not only for selective recognition but also as eco-friendly catalyst.


Direct Clostridioides difficile ribotyping from stool using capillary electrophoresis.
Clostridioides difficile(C. difficile) genotyping is essential for surveillance of emerging strains, transmissions, and outbreak investigations, but culture is lengthy and may not be routinely performed, which necessitates culture-independent genotyping methods. We aimed to develop a direct from stool C. difficile PCR ribotyping algorithm using capillary electrophoresis. Ribotypes were generated directly from 66.8% of stools with 33.2% requiring broth enrichment. 16S and tcdB cycle thresholds (Ct) were significantly lower (P< 0.001) in directly ribotyped stools compared to enriched stools, and Ct correlated with direct ribotyping (area under the curve: 0.97 and 0.96, respectively). Direct and isolate ribotypes were 94.7% concordant. Mixed C. difficile ribotypes were presumptively identified in 14 (7.5%) samples with 12 (6.4%) mixtures confirmed. We have developed a rapid PCR ribotyping algorithm allowing for direct C. difficile genotyping from stool using capillary electrophoresis with occasional detection of mixed C. difficile populations in stool, which is a limitation of conventional isolate genotyping.


Protein immobilization and fluorescence quenching on polydopamine thin films.
Mussel inspired polydopamine (PDA) film has attracted great interest as a versatile functional coating for biomolecule immobilization in various bio-related devices. However, the details regarding the interaction between a protein and PDA film remain unclear. Particularly, there is very limited knowledge regarding the protein immobilization on PDA film, even though it is of essential importance in various fields. The situation is even more complicated if considering the fact that quite a number of approaches (e.g., different oxidizing reagent, buffer pH, grown time, grown media, etc.) have been developed to grow PDA films. In this work, protein attachment on PDA film was systematically investigated by using the real-time and label-free surface plasmon resonance (SPR) technique. The kinetics of protein-PDA interaction was explored and the influence of buffer pH and deposition media on the protein attachment was studied. Fluorescent protein microarray was further printed on PDA-coated glass slides for quantitative investigations and together with SPR data, the interesting fluorescence quenching phenomenon of PDA film was revealed. This work may deepen our understanding on the PDA-protein interaction and offer a valuable guide for efficient protein attachment on PDA film in various bio-related applications.


Thiourea-based spacers in potent divalent inhibitors of Pseudomonas aeruginosa virulence lectin LecA.
A new divalent highly potent inhibitor of the Pseudomonas aeruginosa lectin and virulence factor LecA was prepared. It contains two thiourea linkages which were found to be in the Z,Z isomeric form. This brings the spacer into an elongated conformation required to bridge the two binding sites, which results in the chelating binding mode responsible for the high potency.


Clinical significance of creative 3D-image fusion across multimodalities [PET+CT+MR] based on characteristic coregistration.
To investigate a registration approach for 2-dimension (2D) based on characteristic localization to achieve 3-dimension (3D) fusion from images of PET, CT and MR one by one. A cubic oriented scheme of"9-point & 3-plane" for co-registration design was verified to be geometrically practical. After acquisiting DICOM data of PET/CT/MR (directed by radiotracer 18F-FDG etc.), through 3D reconstruction and virtual dissection, human internal feature points were sorted to combine with preselected external feature points for matching process. By following the procedure of feature extraction and image mapping, "picking points to form planes" and "picking planes for segmentation" were executed. Eventually, image fusion was implemented at real-time workstation mimics based on auto-fuse techniques so called "information exchange" and "signal overlay". The 2D and 3D images fused across modalities of [CT+MR], [PET+MR], [PET+CT] and [PET+CT+MR] were tested on data of patients suffered from tumors. Complementary 2D/3D images simultaneously presenting metabolic activities and anatomic structures were created with detectable-rate of 70%, 56%, 54% (or 98%) and 44% with no significant difference for each in statistics. Currently, based on the condition that there is no complete hybrid detector integrated of triple-module [PET+CT+MR] internationally, this sort of multiple modality fusion is doubtlessly an essential complement for the existing function of single modality imaging.


Elliptical metallic rings-shaped fractal metamaterial absorber in the visible regime.
Achieving the broadband response of metamaterial absorbers has been quite challenging due to the inherent bandwidth limitations. Herein, the investigation was made of a unique kind of visible light metamaterial absorber comprising elliptical rings-shaped fractal metasurface using tungsten metal. It was found that the proposed absorber exhibits average absorption of over 90% in the visible wavelength span of 400-750 nm. The features of perfect absorption could be observed because of the localized surface plasmon resonance that causes impedance matching. Moreover, in the context of optoelectronic applications, the absorber yields absorbance up to ~ 70% even with the incidence obliquity in the range of 0°-60° for transverse electric polarization. The theory of multiple reflections was employed to further verify the performance of the absorber. The obtained theoretical results were found to be in close agreement with the simulation results. In order to optimize the results, the performance was analyzed in terms of the figure of merit and operating bandwidth. Significant amount of absorption in the entire visible span, wide-angle stability, and utilization of low-cost metal make the proposed absorber suitable in varieties of photonics applications, in particular photovoltaics, thermal emitters and sensors.


[Applying active learning in pharmaceutical studies by the way of team-based learning].
Currently, the most used pedagogical approach in pharmaceutical studies is transmissive teaching, in which students usually adopt a passive attitude. The aim of this article is to present the main characteristics of active learning, and to point how it can be integrated in pharmaceutical studies by the way of team-based learning. According to its different characteristics, team-based learning is an authentic method of active learning. This pedagogical method includes a preparation time, usually individually performed, and followed by the group work itself, in which the case method is the most frequently used. Team-based learning also allows students to work in interdisciplinarity and facilitates interactions between peers. The wide majority of the studies demonstrate that students acclaim this pedagogical approach as compared to more conventional teaching methods, and that their satisfaction increases their commitment in the tasks proposed in the course. Even though the results of studies concerning the impact of team-based learning on student retention and scores in tests are somewhat discordant, this pedagogical approach allows the acquisition or the reinforcement of skills that will be useful to students in their future professional life, like collaborative work and confidence in their ability to complete a task.


Anti-HIV-1 activity of pyrryl aryl sulfone (PAS) derivatives: synthesis and SAR studies of novel esters and amides at the position 2 of the pyrrole nucleus.
A SAR study has been performed in order to evaluate how much the ester function could be a determinant for the anti-human immunodeficiency virus type-1 activity of pyrryl aryl sulfones (PASs), a potent family of non-nucleoside reverse transcriptase (RT) inhibitors discovered in the last years. Twenty-three new esters were prepared with the aim to enhance the inhibitory potency of 4a and 4c, two PAS agents endowed with good activity (EC50 = 0.14 microM) and deprived of cytotoxicity up to >200 microM. None of test derivatives was as potent as 4a and 4c and lacked of selectivity due to their higher cytotoxicity (compounds 22-25). Antiviral activity correlate with an ester ramified chain.


Determination of nifedipine in human plasma by solid-phase extraction and high-performance liquid chromatography: validation and application to pharmacokinetic studies.
Nifedipine, a dihydropyridine calcium channel antagonist, is widely used in the treatment of hypertension and other cardiovascular disorders. A simple, rapid, sensitive, precise and accurate HPLC method, using solid-phase extraction, for the quantitation of nifedipine in human plasma was developed and validated. The calibration graphs were linear in the 5-400 ng/ml concentration range (r>0.999). Recovery for nifedipine was greater than 93.9% and for internal standard nitrendipine was 96.1%. Intra-day and inter-day precision ranged from 1.4 to 4.2 and 3.9 to 5.6%, respectively. Intra-day and inter-day accuracy was ranged from 94.5 to 98.0 and 93.1 to 98.0%, respectively. The method was not interfered with by other plasma components and was applied for the determination of nifedipine in pharmacokinetic study after single oral administration of 10 mg nifedipine to 18 healthy male subjects.


Inhaler device feature preferences among patients with obstructive lung diseases: A systematic review and meta-analysis.
Bronchodilators administered through inhalation devices are the mainstay treatment for patients with obstructive lung diseases. Patients do not view devices as interchangeable. This systematic review and meta-analysis examined device feature preferences among patients with obstructive lung diseases treated with handheld inhalers. PubMed, EMBASE, PsycINFO, Cochrane, and Google Scholar were searched to identify publications between 2010 and 2019 that met the following criteria:A manual search extended the study period from 2001 to 2019. Random-effects models were used to generate pooled mean effect sizes and 95% confidence intervals (CIs) for preferred device features. Heterogeneity was measured by the I statistic. Nineteen studies (n = 11,256) were included in this meta-analysis. Average age ranged from 50.4 to 74.3 years. The majority of patients were male (57%) and had chronic obstructive pulmonary disease (92%).Patients preferred the following device features: CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS:: Adults with obstructive lung diseases preferred small inhaler devices that were portable, durable, perceived as easy to use, and fast in medication administration. Healthcare providers should give due consideration to the patient's device feature preferences when developing a treatment plan that prescribes an inhalation device.


Messenger ribonucleic acid for the mouse decidual prolactin is present and induced during in vitro decidualization of endometrial stromal cells.
Although decidualization of endometrial stromal cells is crucial for blastocyst implantation and maintenance of pregnancy, its complex mechanism still remains largely unknown. To explore the mechanisms of decidualization, we have established an in vitro decidualization model in mice. Through extraction of mRNA from endometrial stromal cells for subsequent reverse transcriptase polymerase chain reaction analysis, we found that two ovarian hormones, 17 beta-estradiol and progesterone, induced morphological changes and expression of mRNA for prolactin (PRL) and the decidual/trophoblastic PRL-related protein (d/tPRP) in cultured mouse endometrial stromal cells. The expression of mRNA for the decidual PRL was also detected in the mouse decidua in vitro. In conclusion, mouse mRNA for decidual PRL is present and induced by estradiol and progesterone through in vitro decidualization of endometrial stromal cells.


Abnormalities of serum potassium concentration in dialysis-associated hyperglycemia and their correction with insulin: a unique clinical/physiologic exercise in internal potassium balance.
The absence of significant losses of potassium in the urine makes dialysis-associated hyperglycemia (DH) a model for the study of the internal potassium balance. Studies of DH have revealed that hyperkalemia is frequent at presentation, insulin infusion is usually the only treatment required, and the magnitude of the decrease in serum potassium concentration (K(+)) during treatment of DH with insulin depends on the starting serum K(+) level, the decreases in serum glucose concentration and tonicity, and the increase in serum total carbon dioxide level. We present an analysis of these findings based on previously studied actions of insulin. Calculations of transcellular potassium shifts based on the combined effects of insulin-the increase in the electrical potential differences (hyperpolarization) of the cell membranes and the correction of the hyperglycemic intracellular dehydration through decrease in serum glucose concentration-produced quantitative predictions of the decrease in serum K(+) similar to the reported changes in serum K(+) during treatment of DH with insulin. The lessons from analyzing serum K(+) changes during treatment of DH with insulin are applicable to other conditions where internal potassium balance is called upon to protect serum K(+), such as the postprandial state. The main questions related to internal potassium balance in DH that await clarification include the structure and function of cell membrane potassium channels, the effect of insulin on these channels, and the mechanisms of feedforward potassium regulation.


Neutron Beam Radiation Therapy: An Overview of Treatment and Oral Complications When Treating Salivary Gland Malignancies.
There is limited information in the literature regarding the oral complications of neutron radiotherapy, with an associated lack of guidelines for their management. The purpose of this study was to review the indications, dosing, prognosis, and oral complications of neutron radiotherapy for salivary gland malignancies. This is a retrospective case series of patients with salivary gland malignancies treated with neutron radiotherapy by the Radiation Oncology Department at the University of Washington from 1997 to 2006. Variables included patient demographics, tumor staging and characteristics, operative treatment, final diagnosis, operative and pathologic findings (ie, perineural invasion, lymph node involvement, and skull base invasion), dosing, complications, and locoregional control and survival rates. Data were extracted from patients' charts and by telephone for follow-up and quality-of-life information. The sample was composed of 140 patients (49% men, 51% women) with a mean age of 53 years (standard deviation, 15 yr). Adenoid cystic carcinoma of the submandibular gland was the most common tumor type and location. Post-treatment trismus occurred in 56%. Acute mucositis and xerostomia occurred in approximately 88 and 89% of patients, respectively. Osteoradionecrosis was reported in 5.7% of patients. The 6-year survival rate was 58% and the 6-year locoregional control was 72%. The current standard neutron dose for head and neck tumors is 1.15 neutron Gray (nGy) 4 times per week for 4 weeks (total, 18.4 nGy), which is an equivalent amount of radiation as the standard 60 to 70 Gy given for 6 to 7 weeks with conventional photon radiation. The 6-year overall survival of 58% found in this study compares favorably to the survival rate reported in the literature for traditional photon radiation treatment of advanced salivary gland tumors. The dental profession should be educated regarding neutron radiotherapy and its indications, dosing methods, and oral complications.


Interhemispheric hypoconnectivity in schizophrenia: fiber integrity and volume differences of the corpus callosum in patients and unaffected relatives.
Changes in hemispheric asymmetry and inter-hemispheric connectivity have been reported in schizophrenia. However, the genetic contribution to these alterations is still unclear. In the current study, we applied an automatic segmentation method to structural MRI and diffusion tensor imaging (DTI) data and examined volume and fiber integrity of the corpus callosum (CC), the main interhemispheric fiber tract, in 16 chronic schizophrenia (SZ) patients, matched first degree relatives and controls. SZ patients and relatives had smaller CC volumes than controls, particularly in the posterior genu, isthmus and splenium. Fractional anisotropy (FA), an indicator of fiber integrity, was reduced in patients and relatives in the whole CC, the inferior genu, the superior genu and the isthmus. Correspondingly, the mean diffusivity (MD) values of the whole CC and the isthmus were higher in patients and their unaffected relatives, indicating decreased compactness and increased intercellular space. Relatives had intermediate values in the volumetric and fiber integrity measurements between patients and controls. Lower CC volume and fiber integrity in SZ patients were associated with more severe auditory hallucinations. These results support the connectivity hypothesis of SZ (Friston, 1998) and particularly highlight the altered interhemispheric connectivity, which appears to be a genetic feature of SZ risk.


Effects of methotrexate on glycosaminoglycan production by scleroderma fibroblasts in culture.
To determine the effects of increasing concentrations of methotrexate on the proliferation and glycosaminoglycan (GAG) synthesis of cultured dermal fibroblasts from patients with scleroderma. Cultured dermal fibroblasts from nine patients with scleroderma and nine normal volunteers were grown for 72 hours in media containing various concentrations of methotrexate. The GAG synthesis in each cell was measured after incubating the fibroblasts with [3H]glucosamine and [35S]sulphate. A negative correlation was found between the concentration of methotrexate and numbers of fibroblasts from patients with scleroderma and normal controls. A positive correlation was found between GAG synthesis in each cell, as measured by [3H]glucosamine and [35S]sulphate incorporation, and increasing methotrexate concentrations in fibroblasts from patients with scleroderma and normal controls. These data indicate increased GAG synthesis in scleroderma and normal fibroblasts with increasing concentrations of methotrexate. Therefore the reported beneficial effect of methotrexate on skin fibrosis in scleroderma is most probably not the result of direct inhibition of GAG synthesis by fibroblasts.


Palliative urinary diversion in patients with advanced cervical cancer.
An analysis of 25 palliative urinary diversions for advanced cervical cancer is presented. The median postoperative survival was 176 days and the mean was 329 days. Indication, methods of surgical procedure and duration of known disease did not strongly influence survival. The patients who were treated with radiation therapy after urinary diversion had a longer median survival. Renal function returned to normal in 7 of 9 azotemic patients. 41% of the patients left the hospital and this group subsequently spent 62% of their remaining survival at home. Guidelines for selection of the patients who might benefit from urinary diversion are discussed.


The association of serum prolidase activity with developmental dysplasia of the hip.
The purpose of this study was to evaluate serum prolidase activity in patients with developmental dysplasia of the hip (DDH). Prolidase enzyme activity was measured spectrophotometrically to pointing out the collagen metabolism. The prolidase activity in patients with DDH was significantly higher than that in the control group (p = 0.002). Furthermore, there was positive correlation between prolidase activity and dysplasia level. Increased serum prolidase activity may have played a role in the presence of DDH. We therefore hypothesized that the increased prolidase activity related to collagen turnover may be associated with etiopathogenesis and/or the progression of the disease.


Short range magnetic exchange interaction favors ferroelectricity.
Multiferroics, where two or more ferroic order parameters coexist, is one of the hottest fields in condensed matter physics and materials science. To search multiferroics, currently most researches are focused on frustrated magnets, which usually have complicated magnetic structure and low magnetic ordering temperature. Here, we argue that actually simple interatomic magnetic exchange interaction already contains a driving force for ferroelectricity, thus providing a new microscopic mechanism for the coexistence and strong coupling between ferroelectricity and magnetism. We demonstrate this mechanism by showing that even the simplest antiferromagnetic insulator like MnO, could display a magnetically induced ferroelectricity under a biaxial strain. In addition, we show that such mechanism also exists in the most important single phase multiferroics, i.e. BiFeO3, suggesting that this mechanism is ubiquitous in systems with superexchange interaction.


Pulmonary tumor thrombotic microangiopathy with cor pulmonale due to desmoplastic small round cell tumor.
A 12-year-old boy presented acutely after an episode of syncope with perioral cyanosis. He died 19 hours after admission due to cor pulmonale as a complication of metastatic desmoplastic small round cell tumor in the lungs with associated tumor thrombotic microangiopathy.


A rapid and sensitive radioimmunohistochemical assay for quantitation of vasopressin in discrete brain regions with an anatomical resolution.
Radioimmunoassay has become a widely used method to study different neuroactive substances from brain tissue extracts, but cannot provide anatomical resolution. Here we describe a simple and sensitive radioimmunohistochemical assay (RIHA) to quantify a peptide, vasopressin (VP), in discrete brain regions of rats with 3-day water deprivation. After decapitation, brains were removed, frozen with dry ice and cut into 14-microns cryostat sections which were then fixed with 4% paraformaldehyde in phosphate-buffered saline. After rinses, tissue sections were stored in a freezer until use. For RIHA, brain tissue sections were pre-incubated, and then incubated with rabbit vasopressin antibody (1:2000 dilution) for 24 h at room temperature. After rinses, sections were incubated with 125I-labeled goat antirabbit IgG (1:200 dilution) for 1 h. Specimens were processed for quantitative autoradiography after rinses and drying. RIHA with aid of a computer-assisted image analysis system revealed that the VP content was significantly reduced in the paraventricular hypothalamic nucleus (PVN) and supraoptic nucleus (SON) of rats with 3-day water deprivation, whereas a parallel in situ hybridization study further demonstrated that VP mRNAs in the PVN and SON were greatly increased. In summary, this experiment demonstrates that RIHA is a simple and powerful tool which is able to detect changes of VP in the hypothalamus of dehydrated rats. Combining this method with in situ hybridization to assess mRNA expression allows assessment of the functional significance of the peptide changes. In this case, dehydration depletes vasopressin and upregulates its synthesis. Therefore, the combined use of RIHA and in situ hybridization should have general applicability to evaluate the functional role of a peptide or neurotransmitter system in response to stimuli in a quantitative way with anatomical resolution.


Prostate-specific antigen kinetics in screen-detected prostate cancer in Japan.
It would be of value to compare the features of prostate cancer detected in various screening series around the world. Recently, some studies have demonstrated the value of pretreatment prostate-specific antigen (PSA) kinetics in predicting the outcome of radical prostatectomy and radiotherapy for men with localized prostate cancer. Therefore, the distribution of PSA velocity (PSAV) or PSA doubling time in screen-detected prostate cancer might be objective parameters to investigate how well each national screening system is working. From 1992 to 2004, 957 men with prostate cancer were detected by screening in Gunma Prefecture, Japan. Of those, 275 men (29%) detected with consecutive screening tests participated in the present study. The PSAV was calculated by the PSA change between the most recent screening test and cancer diagnosis and also by linear regression analysis. The PSA doubling time was also calculated for 146 men who underwent screening at least three times. The median PSAV was 1.3 ng/mL/yr in those with Stage T1cN0M, 1.1 ng/mL/yr in those with T2N0M0, and 2.1 ng/mL/yr in those with T3N0M0. The percentage of men with a PSAV (linear regression analysis) greater than 2.0 ng/mL/yr was 13%, 12%, and 49% in men with clinical Stage T1cN0M0, T2N0M0, and T3N0M0, respectively. The median PSA doubling time was 57.1, 51.7, and 28.0 months for those with T1cN0M0, T2N0M0, and T3N0M0, respectively. Patients with prostate cancer with aggressive features are still detected in the population-based screening system in Japan. Even in Japan, where PSA screening is perhaps the most widespread among Asian countries, the screening system might be still immature compared with the systems in the United States and Western Europe.


Needle extraction device for rapid and quantitative gas chromatographic determination of volatile chlorinated hydrocarbons and benzene in soil.
A rapid and quantitative analytical method for volatile chlorinated hydrocarbons and benzene, such as chloroethylene, dichloromethane, and carbon tetrachloride, in soil samples was developed using a needle-type extraction device packed with activated carbon-based adsorbents. The soil sample of 10 g was placed in a glass cartridge and fixed with quartz wool to make sure not to move during the extraction process. After the internal standard solution was spiked into the soil sample, the cartridge was sealed with a polytetrafluoroethylene adapter, and the needle extraction device was used to collect the headspace gas. Analytes were purged from the soil sample and collected on the adsorbent, while pure nitrogen gas was introduced to the soil sample during the sample collection. The sampling time was approximately 10 min for 100 mL of gas sampling. After the sampling process, the extraction needle was directly inserted into a heated injection port of gas chromatography equipped with flame ionization detector (GC-FID) or that hyphenated to mass spectrometry (GC-MS). The extracted VOCs were thermally desorbed and simultaneously introduced to a GC capillary column for subsequent separation and determination. The limit of detection and limit of quantification of all the examined analytes was less than 20 and 60 ng/10 g soil in GC-FID and less than 2.5 and 7.5 ng/10 g soil in the case of GC-MS. Linear calibration curves were obtained up to 10 μg/10 g soil for all the analytes. The relative standard deviations of the ratio of analyte peak area and internal standard peak area were less than 10%. The recovery and sensitivity of analytes confirmed in the proposed method were compared with that obtained with a standard method conventionally employed.


[Functional disorders of the lower urinary tract in children].
Functional disorders of the lower urinary tract as well as vesicoureteral reflux involved in the disease complex of urinary tract infection/permanent renal parenchymal damage can be considered predisposing or risk factors. Two main forms can be distinguished, i.e., unstable bladder and dysfunctional voiding, while transitional forms between the two exist. Functional disorders of the lower urinary tract obstruct spontaneous resolution of vesicoureteral reflux. They are found in about 50% of cases in all children with urinary tract infection and are associated with an increased risk of developing renal parenchymal scars. They are observed during the newborn period up to school age. In the first few months of life, particularly boys with bilateral high-grade reflux and congenital renal parenchymal damage are affected. At later ages girls are also affected, but in this age group bladder instability predominates. Incontinence as the leading clinical symptom appears in approximately 70% of all cases and is closely correlated with chronic constipation. Imaging procedures in addition to urodynamic methods are of decisive importance for diagnosis and treatment, but noninvasive approaches such as sonography should be given preference.


Neuropathic bladder dysfunction in patients with ossification of the posterior longitudinal ligament.
Ossification of the posterior longitudinal ligament (OPLL) may cause neuropathic bladder dysfunction due to spinal cord involvement. OPLL, unlike a traumatic spinal cord lesion, progresses insidiously and sometimes affects longer cord segments. As the manifestation of bladder dysfunction may depend on the development of OPLL, we studied the relationship between bladder function and roentgenographic changes in the spinal canals of OPLL patients. Eighteen surgical candidates (11 males and 7 females, 34 to 85 years old) were studied urodynamically. Sixteen underwent CO2-filling cystometry, uroflowmetry and measurement of their residual urine volume. Cystometry was omitted in the remaining 2 patients. The vertical extent of OPLL and the degree of stenosis in the spinal canal was estimated by x-ray films and CT. The cystometric study revealed detrusor hyperreflexia in 2 patients and areflexic or underactive detrusors in 5 patients. Intermittent flows or considerable amounts of residual urine were also observed in the arefilexia/underactive group. Uroflowmetry showed a normal flow with little residual urine in both patients in whom cystometry was omitted. Bladder sensation was maintained in all patients. The occurrence of abnormal detrusor activity had no relationship to the degree of canal stenosis, while the occurrence of an areflexic or underactive detrusor correlated with the vertical extent of OPLL. Although detrusor hyperreflexia is common in an upper spinal cord lesion, attention should also be paid to the development of detrusor underactivity in patients with a wide vertical extent of OPLL.


Circulating inflammation-resolving lipid mediators RvD1 and DHA are decreased in patients with acutely symptomatic carotid disease.
Efficient biomarkers for early prediction and diagnosis of an acutely symptomatic carotid plaque rupture event are currently lacking, impairing the ability to diagnose and treat patients with an acute plaque rupture events in a timely fashion. Resolvins are endogenous inflammation-resolving lipid mediators that are induced by inflammatory insults. We hypothesized that resolvin and other lipid profiles in sera likely mark the process towards plaque rupture. Circulating lipids associated with plaque rupture events were quantitatively profiled via targeted mediator-lipidomics using ultraperformance liquid chromatography tandem mass spectrometry in patients with acutely symptomatic and asymptomatic carotid disease. Resolvin D1 (RvD1, 82 ± 11pM vs. 152 ± 17pM, p = 0.001) and docosahexaenoic acid (DHA) (0.052 ± 0.007µM versus 0.076 ± 0.008µM, p = 0.025) levels are decreased in the sera of patients presenting with an acutely symptomatic carotid plaque rupture event (n = 21) compared to patients with asymptomatic (n = 24) high-grade carotid stenosis. Circulating arachidonic acid (AA) levels, however, were higher (0.429 ± 0.046µM versus 0.257 ± 0.035µM, p < 0.01) in acutely symptomatic compared to asymptomatic carotid patients. ROC curve analysis demonstrates that the serum ratio AA:RvD1 (AUC 0.84, sensitivity 0.71, specificity 0.92) and AA:DHA (AUC 0.86, sensitivity 0.90, specificity 0.71) are biomarkers for the risk of atherosclerotic plaque rupture. A circulating pro-inflammatory lipid profile, characterized by high AA:RvD1 and AA:DHA, is associated with acutely symptomatic carotid disease and stroke.


Application of systems biology to identify pharmacological mechanisms of thrombotic microangiopathy evoked by combined activated prothrombin complex concentrate and emicizumab.
Emicizumab is a bispecific monoclonal antibody that substitutes for the function of missing or deficient factor VIII (FVIII) in people with hemophilia A (PwHA). Long-term safety and efficacy of emicizumab have been demonstrated in several clinical trials. Nevertheless, in the first of these, three cases of thrombotic microangiopathy (TMA) occurred in PwHA treated with emicizumab receiving high doses of activated prothrombin complex concentrate (aPCC), a bypassing agent used for treating breakthrough bleeds when FVIII neutralizing antibodies (inhibitors) make FVIII replacement ineffective. The aim of the present work is to offer a method to elucidate the pathophysiological and pharmacological mechanisms involved in this treatment-induced TMA. Systems biology and machine learning-based Therapeutic Performance Mapping System is a validated in silico technology that allowed us to construct models of potential mechanisms behind induced TMA. Two drug combinations were modeled and assessed: emicizumab plus aPCC and emicizumab plus recombinant activated factor VII (another bypassing agent). Our models showed that both combinations were related to activation of the coagulation cascade. However, mechanisms involved mainly in platelet activation and possibly in complement activation were detected only for emicizumab plus aPCC, potentially explaining the occurrence of TMA only in this combination.


Assessing the relationship between latent inhibition and the partial reinforcement extinction effect in autoshaping with rats.
Results from a variety of independently run experiments suggest that latent inhibition (LI) and the partial reinforcement extinction effect (PREE) share underlying mechanisms. Experiment 1 tested this LI=PREE hypothesis by training the same set of rats in situations involving both nonreinforced preexposure to the conditioned stimulus (LI stage) and partial reinforcement training (PREE stage). Control groups were also included to assess both LI and the PREE. The results demonstrated a significant, but negative correlation between the size of the LI effect and that of the PREE. Experiment 2 extended this analysis to the effects on LI and the PREE of the anxiolytic benzodiazepine chlordiazepoxide (5 mg/kg, i.p.). Whereas chlordiazepoxide had no effect on LI, it delayed the onset of the PREE. No evidence in support of the LI=PREE hypothesis was obtained when these two learning phenomena were compared within the same experiment and under the same general conditions of training.