Activation of mouse Pi-class glutathione S-transferase gene by Nrf2(NF-E2-related factor 2) and androgen.
The Pi-class glutathione S-transferases (GSTs) play pivotal roles in the detoxification of xenobiotics, carcinogenesis and drug resistance. The mechanisms of regulation of these genes during drug induction and carcinogenesis are yet to be elucidated. Recently, Nrf2 (NF-E2-related factor 2; a bZip-type transcription factor) knockout mice were shown to display impaired induction of Pi-class GST genes by drugs. It is known that the mouse Pi-class GST gene GST-P1 is expressed predominantly in the male liver, and is regulated by androgen. To determine whether Nrf2 and the androgen receptor regulate GST-P1 directly, we analysed the molecular mechanism of activation of this gene by these factors. The promoter of the GST-P1 gene was activated markedly by Nrf2 in transient transfection analyses. Gel mobility shift assay and footprinting analyses revealed three Nrf2 binding sites: one at the proximal and two at distal elements, located at positions -59, -915 and -937 from the cap site. The fifth intron of the GST-P1 gene contains the androgen-responsive region. Multiple androgen receptor binding sites are clustered within a 500 bp region of this intron. The whole fragment contains a minimum of seven androgen receptor binding sites, which collectively display strong androgen-dependent enhancer activity. However, on division into small fragments containing two or three elements each, individual enhancer activities were dramatically decreased. This suggests that multiple elements work synergistically as a strong androgen-responsive enhancer. Our findings indicate that Nrf2 and the androgen receptor directly bind to and activate the mouse GST-P1 gene.


Evolution of cutaneous leishmaniasis in the last 30 years in a tertiary hospital of the European Mediterranean coast.
Although with a lower incidence than in other geographic areas, leishmaniasis is also endemic on the European Mediterranean coast. However, there are few studies on the clinical features of cutaneous lesions of leishmaniasis in Europe. Our objective was to review the clinical features of cutaneous leishmanial lesions in our European Mediterranean population in the last 30 years and compare the clinical features of immunosuppressed and nonimmunosuppressed patients. The clinical features of cutaneous lesions of leishmaniasis diagnosed between 1987 and 2016 at Bellvitge Hospital in Barcelona, Spain, were retrospectively analyzed. Cutaneous lesions of leishmaniasis were diagnosed in 68 patients (40 male and 28 female, mean age 53.60 years, SD 19.68). Thirteen patients were immunosuppressed because of acquired immune deficiency syndrome (AIDS) (7), renal transplantation (1), lymphoma (1), and anti-TNF agents (4). Our immunosuppressed patients had more lesions (3.33 vs. 1.80, P = 0.021), with greater maximum diameter (33.00 vs. 13.33 mm, P = 0.001), and their lesions were more frequently disseminated (P = 0.008). Visceral leishmaniasis was observed only in immunosuppressed patients. Patients treated with anti-TNF drugs developed unusually large skin lesions with crusted, eroded surfaces and without a tendency to spontaneous remission. With the widespread use of anti-TNF agents, an increase in severe forms of leishmaniasis can be expected. The development of persistent, crusted, or eroded erythematous-brownish plaques in patients treated with anti-TNF drugs who live or had traveled to endemic areas of Leishmania infection warrants consideration of a diagnosis of cutaneous leishmaniasis.


A new lectin family with structure similarity to actinoporins revealed by the crystal structure of Xerocomus chrysenteron lectin XCL.
A newly defined family of fungal lectins displays no significant sequence similarity to any protein in the databases. These proteins, made of about 140 amino acid residues, have sequence identities ranging from 38% to 65% and share binding specificity to N-acetyl galactosamine. One member of this family, the lectin XCL from Xerocomus chrysenteron, induces drastic changes in the actin cytoskeleton after sugar binding at the cell surface and internalization, and has potent insecticidal activity. The crystal structure of XCL to 1.4 A resolution reveals the architecture of this new lectin family. The fold of the protein is not related to any of the several lectin folds documented so far. Unexpectedly, the structure similarity is significant with actinoporins, a family of pore-forming toxins. The specific structural features and sequence signatures in each protein family suggest a potential sugar binding site in XCL and a possible evolutionary relationship between these proteins. Finally, the tetrameric assembly of XCL reveals a complex network of protomer-protomer interfaces and generates a large, hydrated cavity of 1000 A3, which may become accessible to larger solutes after a small conformational change of the protein.


Fatty Acid Fueled Transmembrane Chloride Transport.
Generation of chemical gradients across biological membranes of cellular compartments is a hallmark of all living systems. Here we report a proof-of-concept prototype transmembrane pumping system in liposomes. The pump uses fatty acid to fuel chloride transport, thus generating a transmembrane chloride gradient. Addition of fatty acid to phospholipid vesicles generates a transmembrane pH gradient (pHin < pHout), and this electrochemical H+ potential is harnessed by an anionophore to drive chloride efflux via H+/Cl- cotransport. Further addition of fatty acid efficiently fuels the system to continuously drive chloride transport against the concentration gradient, up to [Cl-]in 65 mM | [Cl-]out 100 mM, and is 1400 times more efficient than using an external fuel. Based on our findings from dissecting the H+/Cl- flux process with the use of different liposomal fluorescence assays, and supported by additional liposome-based 13C NMR and DLS studies; we proposed that the presence of an anionophore can induce asymmetric distribution of fatty acid, and contribute to another Cl- flux mechanism in this system.


Emerging role of microRNAs in cancer stem cells: Implications in cancer therapy.
A small subset of cancer cells that act as tumor initiating cells or cancer stem cells (CSCs) maintain self-renewal and growth promoting capabilities of cancer and are responsible for drug/treatment resistance, tumor recurrence and metastasis. Due to their potential clinical importance, many researchers have put their efforts over decades to unravel the molecular mechanisms that regulate CSCs functions. MicroRNAs (miRNAs) which are 21-23 nucleotide long, endogenous non-coding RNAs, regulate gene expression through gene silencing at post-transcriptional level by binding to the 3'-untranslated regions or the open reading frames of target genes, thereby result in target mRNA degradation or its translational repression and serve important role in several cellular, physiological and developmental processes. Aberrant miRNAs expression and their implication in CSCs regulation by controlling asymmetric cell division, drug/treatment resistance and metastasis make miRNAs a tool of great therapeutic potential against cancer. Recent advancements on the biological complexities of CSCs, modulation in CSCs properties by miRNA network and development of miRNA based treatment strategies specifically targeting the CSCs as an attractive therapeutic targets for clinical application are being critically analysed.


Case report. Aphasia in thalamic stroke: CT stereotactic localization.
We present a patient in whom dysphasia followed suddenly upon an apparently discrete thalamic infarct proven by computed tomography (CT). Detailed psychometric data of the patient's speech and memory disorder obtained during the acute and chronic stages were correlated with the evaluation of a focal thalamic lesion as demonstrated by serial CT. The findings and deductions drawn from selective reports in the literature form the basis of this presentation.


Prognostic significance of FOXM1 in oral squamous cell carcinoma patients treated by docetaxel-containing regimens.
Forkhead box protein M1 (FOXM1) is an oncoprotein that is involved in cell proliferation, differentiation and aging, and overexpression of FOXM1 is thought to be associated with the development and progression of various types of cancer. The expression of FOXM1 was retrospectively examined in tumor tissues taken from 56 oral squamous cell carcinoma (OSCC) patients by immunohistochemical staining. All of these patients received docetaxel (Doc)-containing regimens as treatments against OSCC. The association between FOXM1 expression and the clinicopathological characteristics and prognosis of these patients was then examined. FOXM1 was expressed in the nucleus and cytoplasm of OSCC tissues samples. There was a significant association between FOXM1 expression in tumor tissues and N classification (P=0.0395), stage (P=0.004), therapeutic efficacy (P=0.0113) and outcome (P=0.0134) of patients. However, FOXM1 expression had no association with patients' sex, age or T classification. Additionally, high expression of FOXM1 in tumor cells was associated with a shorter overall survival (P=0.0257) of patients. Multivariate analysis also revealed that elevated expression of FOXM1 was a predictor of patients' poor survival (P=0.0327). The results suggested that high expression of FOXM1 in OSCC tumors may result in reduced therapeutic effects and poor clinical outcomes of patients receiving Doc-based treatment regimens.


Genome-wide allelic methylation analysis reveals disease-specific susceptibility to multiple methylation defects in imprinting syndromes.
Genomic imprinting is the parent-of-origin-specific allelic transcriptional silencing observed in mammals, which is governed by DNA methylation established in the gametes and maintained throughout the development. The frequency and extent of epimutations associated with the nine reported imprinting syndromes varies because it is evident that aberrant preimplantation maintenance of imprinted differentially methylated regions (DMRs) may affect multiple loci. Using a custom Illumina GoldenGate array targeting 27 imprinted DMRs, we profiled allelic methylation in 65 imprinting defect patients. We identify multilocus hypomethylation in numerous Beckwith-Wiedemann syndrome, transient neonatal diabetes mellitus (TNDM), and pseudohypoparathyroidism 1B patients, and an individual with Silver-Russell syndrome. Our data reveal a broad range of epimutations exist in certain imprinting syndromes, with the exception of Prader-Willi syndrome and Angelman syndrome patients that are associated with solitary SNRPN-DMR defects. A mutation analysis identified a 1 bp deletion in the ZFP57 gene in a TNDM patient with methylation defects at multiple maternal DMRs. In addition, we observe missense variants in ZFP57, NLRP2, and NLRP7 that are not consistent with maternal effect and aberrant establishment or methylation maintenance, and are likely benign. This work illustrates that further extensive molecular characterization of these rare patients is required to fully understand the mechanism underlying the etiology of imprint establishment and maintenance.


Axial patterning in the developing vertebrate inner ear.
Axial patterning in the vertebrate inner ear has been studied for over eighty years, and recent work has made great progress towards an understanding of the molecular mechanisms responsible for establishing asymmetries about the otic axes. Tissues extrinsic to the ear provide sources of signalling molecules that are active early in development, at or before otic placode stages, while intrinsic factors interpret these signals to establish and maintain axial pattern. Key features of dorsoventral otic patterning in amniote embryos involve Wnt and Fgf signalling from the hindbrain and Hh signalling from midline tissues (notochord and floorplate). Mutual antagonism between these pathways and their downstream targets within the otic epithelium help to refine and maintain dorsoventral axial patterning in the ear. In the zebrafish ear, the same tissues and signals are implicated, but appear to play a role in anteroposterior, rather than dorsoventral, otic patterning. Despite this paradox, conservation of mechanisms may be higher than is at first apparent.


Baby Friendly Hospital Initiative practices and breast feeding duration in a cohort of first-time mothers in Adelaide, Australia.
to investigate the relationship between adherence to six of the Baby Friendly Hospital Initiative (BFHI) Ten steps to successful breast feeding and the duration of breast feeding in first-time mothers. a prospective study to assess the duration of breast feeding up to 6 months postpartum. Survival analysis techniques (Kaplan-Meier curves and Cox proportional hazard models) were used to interpret the data. 317 women who had given birth to their first baby (at term) in a large teaching maternity hospital in Adelaide, South Australia, during the period March to November 2003. ignoring all other factors, we found that women whose babies received a bottle feed, used a pacifier or dummy, or who used a nipple shield during their postnatal stay, were at significantly greater risk of weaning (p0.05). After adjusting for socio-demographic variables, self-efficacy, intended duration of breast feeding, and method of delivery, the results unexpectedly showed that the only significant predictor of early weaning was breast feeding on demand. However, a composite variable indicating use of one or more of nipple shields, a dummy or bottle feeds while in hospital resulted in a significantly greater risk of weaning (p=0.05). socio-demographic and cultural factors may be more important determinants of the duration of breast feeding than some of the very specific hospital practices targeted in the Ten steps to successful breast feeding. From a public health perspective, we may influence the duration of breast feeding through better post-discharge support services, or through interventions that improve attitudes to breast feeding in specific socio-cultural and economic groups.


In vitro antifungal activity of several antimicrobial compounds against Penicillium expansum.
Fungicides used in the prevention and control of mold rots in stored apples are subjected to legal, social, and biological limitations. The aim of this study was to find an alternative to postharvest fungicides currently used in the prevention and control of blue mold rot caused by Penicillium expansum in apples. For this purpose, the antimicrobial activity and MIC of several substances against P. expansum were evaluated in vitro using different end point methods: agar diffusion assay, volatility method, and agar dilution and broth dilution MIC assays. Most of the substances tested are common food ingredients and have a recognized antimicrobial activity. Essential oils, such as thymol, eugenol, citral and cineole, vanillin, sodium hypochlorite, acetic acid, potassium sorbate, and hydrogen peroxide, were the substances evaluated. Thymol and citral were the essential oil components that showed the greatest inhibitory effects. The effectiveness of 5 and 10% hydrogen peroxide in growth inhibition of P. expansum in the agar diffusion assay was total, and its MIC as determined by the agar and broth dilution assays was less than 0.025%. These results indicate that the application of small quantities of hydrogen peroxide to the apple skin might be an alternative to fungicides in the elimination of P. expansum.


A molecular marker of disease activity in autoimmune liver diseases with histopathological correlation; FoXp3/RORγt ratio.
Autoimmune liver diseases (AILDs) encompass a group of diseases with variable clinicopathological manifestations. Th17 and Treg cells have roles in the pathogenesis of AILDs with a balance shifted towards a relative increase in activity of the Th17 cells. In this study, the balance between the transcription factors of Treg and Th17 cells (FoXp3 and RORγt) was sought as a molecular marker of disease activity and to highlight the pathogenesis. The peripheral blood samples of 46 treatment-naive patients were collected and RNA was extracted. Real time PCR was performed and the ratio of gene expression was calculated. Histopathology of 18 patients was obtained and the activity score of these biopsies were also corroborated with their respective molecular (FoXp3/RORγt) (FRGT=FoXp3-ROR Gamma T) ratio. The FRGT ratio in healthy individuals was close to 1 and in disease the ratio changed significantly. This ratio (FRGT) was not significantly different in different varieties of AILD or in adult or paediatric form of the disease. However, the ratio remained consistently below 1 (mean 0.3) in acute disease and high (mean 224.7) in chronic or asymptomatic form of the disease (p < 0.001). The histopathological activity score also significantly correlated with the ratio. This signified the relative excess of Th17 (RORγt) in active disease as compared to Treg (FoXp3) and the reverse in chronic form. This ratio can be an important peripheral molecular marker to assess the disease activity without the necessity of performing a liver biopsy.


Hepatic dearterialisation induces glucose intolerance and inhibits insulin secretion in patients with liver malignancy.
Intermittent hepatic dearterialisation is used in palliation of liver malignancy. In rats hepatic dearterialisation is accompanied by glucose intolerance and impaired insulin secretion, but it is not known if similar effects occur in man. Six patients with nonresectable liver malignancy were subjected to an intravenous glucose (50 g) challenge before dearterialisation (control challenge), during a 1-hour dearterialisation period, and in the immediate reperfusion phase after the end of 1-hour dearterialisation. Insulin secretion, as judged by the plasma levels of both insulin and C peptide, was inhibited during the 1-hour dearterialisation, while the glucose elimination rate was delayed (both p less than 0.03). Contrastingly, in the immediate reperfusion phase both insulin secretion and glucose elimination did not differ from values after the control challenge. We therefore conclude that hepatic dearterialisation in patients with liver malignancy is accompanied by inhibited insulin secretion and glucose intolerance, both readily reversible.


Molecular mechanism of therapeutic approaches for human gastric cancer stem cells.
Gastric cancer (GC) is a primary cause of cancer-related mortality worldwide, and even after therapeutic gastrectomy, survival rates remain poor. The presence of gastric cancer stem cells (GCSCs) is thought to be the major reason for resistance to anticancer treatment (chemotherapy or radiotherapy), and for the development of tumor recurrence, epithelial-mesenchymal transition, and metastases. Additionally, GCSCs have the capacity for self-renewal, differentiation, and tumor initiation. They also synthesize antiapoptotic factors, demonstrate higher performance of drug efflux pumps, and display cell plasticity abilities. Moreover, the tumor microenvironment (TME; tumor niche) that surrounds GCSCs contains secreted growth factors and supports angiogenesis and is thus responsible for the maintenance of the growing tumor. However, the genesis of GCSCs is unclear and exploration of the source of GCSCs is essential. In this review, we provide up-to-date information about GCSC-surface/intracellular markers and GCSC-mediated pathways and their role in tumor development. This information will support improved diagnosis, novel therapeutic approaches, and better prognosis using GCSC-targeting agents as a potentially effective treatment choice following surgical resection or in combination with chemotherapy and radiotherapy. To date, most anti-GCSC blockers when used alone have been reported as unsatisfactory anticancer agents. However, when used in combination with adjuvant therapy, treatment can improve. By providing insights into the molecular mechanisms of GCSCs associated with tumors in GC, the aim is to optimize anti-GCSCs molecular approaches for GC therapy in combination with chemotherapy, radiotherapy, or other adjuvant treatment.


Expression patterns of prune2 is regulated by Notch and retinoic acid signaling pathways in the zebrafish embryogenesis.
PRUNE2 has been identified as a susceptible gene for Alzheimer's disease and a marker for leiomyosarcomas. Isoforms of Prune2 regulate neuronal cell differentiation and synaptogenesis. Although expression pattern of Prune2 has been reported in the murine brain, its expression patterns and regulation along vertebrate embryogenesis remain to be further investigated. We thus defined the expression profiles and transcriptional regulation of prune2 in zebrafish embryos. prune2 exhibits maternal expression, but is increased in later embryonic stages, and expressed in the telencephalon, epiphysis cluster, nucleus of the tract of the post optic commissure, spinal cord, cerebellum, tegmentum, anterior lateral line ganglion, posterior lateral line ganglion and rhombomeres 2 through 5. Two color WISH with a post-mitotic neuron specific marker, huC defined that prune2 is expressed in the post mitotic neurons. The level of prune2 transcripts is upregulated in Notch signaling homozygous mutant, mib1-/-(mibta52b), indicating that Notch signaling regulates transcription of prune2. Interestingly, in silico analysis of prune2 promoter found retinoic acid (RA) response elements (AGGTCAcaTGACCA) located at -3 to -16 relative to the first exon. It turned out that RA signaling altered the expression pattern of prune2 in the hindbrain. We further propose that Prune2 might be a putative regulator for CNS development in zebrafish embryogenesis.


Detection and isolation of Campylobacter species other than C. jejuni from children with Crohn's disease.
The presence of Campylobacter species other than Campylobacter jejuni and antibodies to Campylobacter concisus in children were investigated. A significantly greater presence of C. concisus and higher levels of antibodies to C. concisus were detected in children with Crohn's disease (CD) than in controls. Campylobacter species other than C. jejuni were isolated from intestinal biopsy specimens of children with CD.


[Surveillance of enteroviruses in the waste water of the Ivory Coast].
Viral water contamination is on the increase all over the world and one is concerned by the dangers to which it exposes health and preoccupied with the search for effective solutions. This study, which was conducted in a year in Côte-d'Ivoire, enabled us to examine the degree of viral pollution from waste water collected from a lagune. It allowed us to follow the distribution of enteroviruses and their fluctuations during the year. 164 waste water samples were tested and 93 enterovirus strains isolated. 67.89% showed polioviruses and 32.11% coxsackieviruses. Seasonal fluctuations of the virus concentration in the lagune water were shown from statistical analysis of the data collected. Concentration was high during the rainy season and definitely lower during the dry season with the rise in water temperature.


Hierarchical approaches for systems modeling in cardiac development.
Ordered cardiac morphogenesis and function are essential for all vertebrate life. The heart begins as a simple contractile tube, but quickly grows and morphs into a multichambered pumping organ complete with valves, while maintaining regulation of blood flow and nutrient distribution. Though not identical, cardiac morphogenesis shares many molecular and morphological processes across vertebrate species. Quantitative data across multiple time and length scales have been gathered through decades of reductionist single variable analyses. These range from detailed molecular signaling pathways at the cellular levels to cardiac function at the tissue/organ levels. However, none of these components act in true isolation from others, and each, in turn, exhibits short- and long-range effects in both time and space. With the absence of a gene, entire signaling cascades and genetic profiles may be shifted, resulting in complex feedback mechanisms. Also taking into account local microenvironmental changes throughout development, it is apparent that a systems level approach is an essential resource to accelerate information generation concerning the functional relationships across multiple length scales (molecular data vs physiological function) and structural development. In this review, we discuss relevant in vivo and in vitro experimental approaches, compare different computational frameworks for systems modeling, and the latest information about systems modeling of cardiac development. Finally, we conclude with some important future directions for cardiac systems modeling.


Impact and prognostic value of 18F-fluorodeoxyglucose positron emission tomography-computed tomography scan in the evaluation of residual head and neck cancer: Single-center experience from Pakistan.
Overview of clinical impact of positron emission tomography-computed tomography (PET-CT) scans in patients with head and neck carcinomas at our center. Retrospective review of posttreatment 18F-fluorodeoxyglucose (18F-FDG) PET-CT scans in patients with head and neck carcinomas with risk of residual disease. Clinical outcome served as the reference standard. This study included 93 patients (65.6% males, mean age: 48.8 years ± 17.2 standard deviation) with squamous cell carcinoma as most frequent histopathology (91.4%). PET-CT scans were performed on average 6 months posttreatment. Diagnostic accuracy, positive predictive value, and negative predictive value of PET-CT for disease were found to be 88%, 88%, and 92%, respectively. A median follow-up of 24 months was available for 91 patients. Kaplan-Meier curves showed significantly higher disease-free survival with negative PET-CT as compared to positive PET-CT (P = 0.01) and maximum standardized uptake values of <5.0 (P = 0.01). FDG PET-CT has diagnostic and prognostic implications in treated patient of head and neck cancers.


Relative exposure to swine animal feeding operations and childhood asthma prevalence in an agricultural cohort.
Large swine animal feeding operations (AFOs) have become the model of livestock production throughout the United States. Epidemiological studies have consistently shown an increase in adverse respiratory symptoms among workers at AFOs. However, the impact on communities surrounding these facilities is still being investigated. We evaluated the association between relative environmental exposure to AFOs and the prevalence of prescribed medication for wheeze and/or childhood asthma in rural Iowa. Demographic and health information on 565 children aged 0-17 was obtained from a previous population-based cohort study while data on the AFOs were collected from publically available tax records. We created a metric of each child's relative environmental exposure to swine CAFOs which incorporated the size of the AFO as well as distance and wind direction. We determined the association between self-reported prescription medication for wheeze and/or self-reported physician diagnosed asthma and relative exposure while controlling for recognized risk factors using correlated logistic regression. The prevalence of childhood asthma in the cohort was 11.0% while 22.7% of children had been previously prescribed medication for wheeze or had a lifetime asthma diagnosis. Children with a larger relative environmental exposure to AFOs had a significantly increased odds of both outcomes (OR=1.51, p=0.014 asthma; OR=1.38, p=0.023 asthma or medication for wheeze). When stratified into exposure quartiles a linear trend was observed with asthma or medication for wheeze as the dependent variable but not with asthma alone. This study is the first to investigate children's cumulative relative exposure to smaller AFOs and adds to the growing volume of literature supporting a link between proximity to swine AFOs and adverse respiratory health.


Mutations in NKX6-2 Cause Progressive Spastic Ataxia and Hypomyelination.
Progressive limb spasticity and cerebellar ataxia are frequently found together in clinical practice and form a heterogeneous group of degenerative disorders that are classified either as pure spastic ataxia or as complex spastic ataxia with additional neurological signs. Inheritance is either autosomal dominant or autosomal recessive. Hypomyelinating features on MRI are sometimes seen with spastic ataxia, but this is usually mild in adults and severe and life limiting in children. We report seven individuals with an early-onset spastic-ataxia phenotype. The individuals come from three families of different ethnic backgrounds. Affected members of two families had childhood onset disease with very slow progression. They are still alive in their 30s and 40s and show predominant ataxia and cerebellar atrophy features on imaging. Affected members of the third family had a similar but earlier-onset presentation associated with brain hypomyelination. Using a combination of homozygozity mapping and exome sequencing, we mapped this phenotype to deleterious nonsense or homeobox domain missense mutations in NKX6-2. NKX6-2 encodes a transcriptional repressor with early high general and late focused CNS expression. Deficiency of its mouse ortholog results in widespread hypomyelination in the brain and optic nerve, as well as in poor motor coordination in a pattern consistent with the observed human phenotype. In-silico analysis of human brain expression and network data provides evidence that NKX6-2 is involved in oligodendrocyte maturation and might act within the same pathways of genes already associated with central hypomyelination. Our results support a non-redundant developmental role of NKX6-2 in humans and imply that NKX6-2 mutations should be considered in the differential diagnosis of spastic ataxia and hypomyelination.


Accelerated separation of GC-amenable lipid classes in plant oils by countercurrent chromatography in the co-current mode.
Triacylglycerols represent the major part (>90%) in most plant oils and have to be eliminated, when the minor compounds such as phytosterols or tocopherols should be analyzed. Here, we used an all liquid-liquid chromatographic technique, countercurrent chromatography (CCC), to fractionate the minor lipids before gas chromatography (GC) analysis. To cover the wide range of polarity of the minor compounds, we used the co-current mode, in which both mobile and stationary phase are pumped through the system. This allowed to elute substances which partitioned almost exclusively in the stationary phase within 90 min. After testing with standard compounds, the method was applied to the separation of sesame oil and sunflower oil samples. The abundant triacylglycerols could be effectively separated from tocopherols, phytosterols, diacylglycerols, and free fatty acids in the samples, and these compounds could be analyzed (after trimethylsilylation) by GC coupled with mass spectrometry. After the enrichment caused by the CCC fractionation, we were also able to identify the tocopherol derivative α-tocomonoenol, which had not been described in sunflower oil before. Also, separation of sesame oil yielded a mixture of the polar compounds sesamin and sesamolin without further impurities.


Fracture recurrence in hip fracture with menopausal hormone therapy versus risedronate: a clinical trial.
An open-label, randomized trial was conducted to examine the effects of risedronate versus menopausal hormone therapy (MHT) in postmenopausal women with recent hip fracture. Among 1165 eligible women, 281 were recruited and randomly assigned to receive oral risedronate (35 mg/week) or percutaneous estradiol gel (1.5 mg/day) plus oral micronized progesterone (100 mg/day) for 4 years. The primary end point was recurrent fracture and the secondary end points were mortality and bone mineral density (BMD). Kaplan-Meier analyses showed no significant differences in fracture recurrence and mortality between the two groups. The incidence of any new fracture per 100 person-years (PY) was 8.63 in the risedronate group and 12.86 in the MHT group (p = 0.180); that of clinical fracture was 4.75 and 6.99, respectively (p = 0.265); and that of asymptomatic vertebral fracture was 4.87 and 5.58, respectively (p = 0.764). The respective incidence of death per 100 PY was 3.58 and 4.40 (p = 0.503). BMD increased comparably at the lumbar spine in both groups. BMD at the total hip did not change in the risedronate group, but increased significantly by 2.8% in the MHT group. MHT might not differ from risedronate in the prevention of secondary fractures and death among postmenopausal women with recent hip fracture.


Individual versus family psychotherapy in managed care: comparing the costs of treatment by the mental health professions.
In an effort to understand how psychotherapy is practiced in the "real world," outpatient claims data were examined to determine the cost of individual and family therapy provided by marital and family therapists, master's nurses, master's social workers, medical doctors, psychologists, or professional counselors. Claims for 490,000 unique persons over 4 years were obtained from CIGNA. Family therapy proved to be substantially more cost-effective than individual or "mixed" psychotherapy. Physicians provided care in the fewest sessions, marital and family therapists had the highest success (86.6%) and lowest recidivism rates (13.4%), and professional counselors were the least costly. Outcomes were overwhelmingly successful, with 85% of patients requiring only one episode of care.


Metabolism and disposition of p-chloroaniline in rat, mouse, and monkey.
The metabolism and disposition of [14C]p-chloroaniline was studied in the male Fisher 344 rat, female C3H mouse, and male rhesus monkey. Greater than 90% of the radiocarbon is eliminated through the urine in all species. A major route of metabolism is orthohydroxylation, whereby 2-amino-5-chlorophenyl sulfate is the major excretion product in all species. Excretion of p-chloro-oxanilic acid and p-chloroglycolanilide, oxidation products of p-chloroacetanilide, is significant in the rat. The appearance of p-chloroacetanilide along with p-chloroaniline in plasma after an oral dose of [14C]p-chloroaniline also was studied in rhesus monkey. In addition to p-chloroacetanilide, 2-amino-5-chlorophenyl sulfate is found to be a major circulating metabolite.


A NEW SPECIES OF MYXOBOLUS (CNIDARIA: MYXOSPOREA: MYXOBOLIDAE) FROM THE GILLS OF CREEK CHUB, SEMOTILUS ATROMACULATUS (CYPRINIFORMES: LEUCISCIDAE: PLAGIOPTERINAE), FROM THE OUACHITA DRAINAGE OF ARKANSAS.
During October and November 2021, 33 creek chubs, Semotilus atromaculatus, were collected from 3 sites in Polk County, Arkansas (Ouachita River drainage), and their gills, gallbladder, fins, integument, musculature, and other major organs were examined for myxozoans. The gills of 9 (27%) were infected with a new myxozoan, Myxobolus fountainae n. sp. Qualitative and quantitative morphological data were from fresh and formalin-fixed preserved spores, while molecular data consisted of a 1918 base pair sequence of the partial small subunit ribosomal RNA gene. Phylogenetic analysis grouped M. fountainae n. sp. with the other leuciscid-infecting myxobolids from North America and within a larger clade of European myxozoans. In addition, histological information is provided on the infection. A previous record of Myxobolus muelleriBütschli, 1882, from the gills and ureters of S. atromaculatus is considered invalid and represents an unknown species. Myxobolus fountainae n. sp. is the only named myxozoan known to infect the gill filaments of S. atromaculatus, whereas Myxobolus pendula (Guilford, 1967) infects the gill arches.


A look inside the courtroom: an analysis of 292 cosmetic breast surgery medical malpractice cases.
Malpractice claims affect the cost and quality of health care. The authors examine litigation in cosmetic breast surgery and identify factors influencing malpractice litigation outcomes. The Westlaw database was searched for jury verdict and settlement reports related to medical malpractice and cosmetic breast surgeries. Cases included for analysis were examined for year, geographic location, patient demographics, procedure performed, alleged injury, causes of action, verdict, and indemnity payments. Of 292 cases, the most common injury sustained was disfigurement (53.1%). Negligent misrepresentation had a 98% greater chance of resolution in favor of the plaintiff (relative risk [RR], 1.98; 95% confidence interval [CI], 1.41-2.79), and fraud had a 92% greater chance of disposition in favor of the plaintiff (RR, 1.92; 95% CI, 1.32-2.80). The most common causes of action cited were negligence (88.7%) and lack of informed consent (43.8%). One hundred sixty-nine (58.3%) cases resulted in favor of the defendant and 121 (41.7%) cases were disposed in favor of the plaintiff; 97 (33.4%) cases resulted in damages awarded and 24 (8.3%) cases resulted in settlement. No significant difference was found between the medians of indemnity payments awarded to plaintiffs ($245 000) and settlements ($300 000). Based on this study, negligent or intentional misrepresentation strongly favors plaintiffs in either awarded damages or settlements in cases of cosmetic breast surgery litigation. This study emphasizes that transparency and adequate communication are at the crux of the physician-patient relationship and are tools by which plastic surgeons may reduce the frequency of litigations, thereby containing health care costs at a minimum.


Pre and post operative medical therapy for endometriosis surgery.
Endometriosis is a common gynaecological condition which affects approximately 10% of women of reproductive age (Askenazi 1997). There is a range of symptoms and most commonly women present with dysmenorrhoea, pelvic pain, infertility or a pelvic mass. Direct visualisation and biopsy during laparoscopy or laparotomy is the gold standard diagnostic test for this condition and enables the gynaecologist to identify the location, extent and severity of the disease. Surgical therapy can be performed concurrently with diagnostic surgery and may include removal (excision) or destruction (ablation) of endometriotic tissue, division of adhesions and removal of endometriotic cysts. Laparoscopic excision or ablation of endometriosis has been shown to be effective in the management of pain in mild-moderate endometriosis. Adjunctive medical treatment pre or post-operatively may prolong the symptom-free interval. To determine the effectiveness of systemic medical therapies used for hormonal suppression before or after surgery for endometriosis, or before and after surgery for endometriosis in the eradication of endometriosis, improvement of symptoms, pregnancy rates and overall tolerability by comparing them with no treatment or placebo. We searched the Cochrane Menstrual Disorders and Subfertility group trials register (searched 10 September 2003), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 3 2003), MEDLINE (January 1966 to September 2003), EMBASE (January 1985 to September 2003) and reference lists of articles. We also contacted researchers in the field. Trials were included if they were randomised controlled trials of the use of systemic medical therapies for hormonal suppression before or after, or before and after surgery for endometriosis. Data extraction and quality assessment was performed independently by using relative risk or weighted mean difference and 95% confidence intervals. Eleven trials were included in the review. One study comparing pre-surgical medical therapy with surgery alone showed a significant improvement in AFS scores in the medical therapy group (WMD -9.60, 95% CI -11.42 to -7.78) but this may or may not be associated with better outcomes for the patients. Post surgical hormonal suppression of endometriosis compared to surgery alone (either no medical therapy or placebo) showed no benefit for the outcomes of pain or pregnancy rates but a significant improvement in disease recurrence (AFS scores (WMD -2.30, 95% CI -4.02 to -0.58)). There were no trials identified in the search that compared hormonal suppression of endometriosis before and after surgery with surgery alone. There is no significant difference between pre surgery hormonal suppression and post surgery hormonal suppression for the outcome of pain in the one trial identified (RR 1.01, 95% CI 0.49 to 2.07). Information concerning AFS scores and ease of surgery was reported only as a descriptive summary so any difference between the groups can not be quantified from the information in the report of this trial. There is insufficient evidence from the studies identified to conclude that hormonal suppression in association with surgery for endometriosis is associated with a significant benefit with regard to any of the outcomes identified. There may be a benefit of improvement in AFS scores with the pre-surgical use of medical therapy. The possible benefit should be weighed in the context of the adverse effects and costs of these therapies.


Nucleosome repeat length and linker histone stoichiometry determine chromatin fiber structure.
To understand how nuclear processes involving DNA are regulated, knowledge of the determinants of chromatin condensation is required. From recent structural studies it has been concluded that the formation of the 30-nm chromatin fiber does not require the linker histone. Here, by comparing the linker histone-dependent compaction of long, reconstituted nucleosome arrays with different nucleosome repeat lengths (NRLs), 167 and 197 bp, we establish that the compaction behavior is both NRL- and linker histone-dependent. Only the 197-bp NRL array can form 30-nm higher-order chromatin structure. Importantly for understanding the regulation of compaction, this array shows a cooperative linker histone-dependent compaction. The 167-bp NRL array displays a limited linker histone-dependent compaction, resulting in a thinner and topologically different fiber. These observations provide an explanation for the distribution of NRLs found in nature.


Ultrastructural observations on a variant of acute promyelocytic leukemia.
A patient with acute leukemia is presented in whom the leukemic cells, as seen by light microscopy were typical promyelocytes. The cells had normal or slightly invaginated nuclei with typical cytoplasmic granules and the diagnosis was confirmed by cytochemistry. The clinical course was rapid and the patient died of disseminated intravascular coagulation and urosepsis within a few days of diagnosis. However, electron microscopic examination showed cells with extremely convoluted and lobulated nuclei with nuclear pockets and cytoplasmic bridges as well as the complete absence of cytoplasmic granules in the majority of the cells. Furthermore, the urine lysozyme (muramidase) was elevated. These findings suggest that the leukemia in this patient may be classified as a hypogranular variant of acute promyelocytic leukemia (APL), with monocytoid ultrastructural appearances.


Serum 25-hydroxyvitamin D and CD4+CD25(+high) FoxP3+ Regulatory T cell as Predictors of Severity of Bronchial Asthma in Children.
Bronchial asthma (BA) is one of the common chronic diseases of childhood. Vitamin D deficiency has been associated with BA. Suppressor regulatory T cells (Treg) are important for the induction, maintenance of immunological tolerance to allergens. This study assessed serum 25-hydroxyvitamin D (vitamin D) and the percentages of CD4+CD25+(high) Foxp3+ Treg, in peripheral blood, as predictors of asthma severity and level of clinical control. The study enrolled 72 children divided equally between asthmatic children (AC) and age and sex matched controls. Diagnostic criteria and level of asthma severity followed the Global Initiative for Asthma guidelines. Serum vitamin D was determined by an immunoassay and the percentages of CD4+CD25+ig Foxp3+ Treg by flow cytometry. Serum vitamin D level and percentage of CD4+CD25+(high) Treg were lower in AC compared to controls (P < 0.001) whereas Fox p3 expression was higher in AC compared to controls, P < 0.001. Serum vitamin D levels were lower in severe asthma compared to mild and moderate forms (P = 0.008) and in uncontrolled attacks compared to partially or completely controlled children. No difference in percentage of Treg in relation to asthma severity and clinical control was observed. Since AC has decreased serum vitamin D with inverse relationship between its levels and asthma severity, we conclude that it can be used to predict severity of asthma.


Family Planning Providers' Experiences and Perceptions of Long-Acting Reversible Contraception in Lilongwe, Malawi.
Less than 2% of Malawian women use long-acting reversible contraception (LARC). We describe experiences of Malawian family planning providers, focusing on LARC. We conducted a mixed-methods study using questionnaires and focus group discussions with providers in Lilongwe. Data were analyzed separately and triangulated. Most (58%) participants saw over 30 patients daily. Only 19% had ever inserted IUC. Qualitative data were complementary; participants noted that LARC provision was important, though hindered by lack of experienced providers, work burden, and low demand. Future efforts to improve LARC access in Lilongwe must address both supply and demand-side barriers.


T cell receptor (TCR)-beta gene recombination: dissociation from cell cycle regulation and developmental progression during T cell ontogeny.
T cell lymphopoiesis involves extensive cell division and differentiation; these must be balanced by export and programmed cell death to maintain thymic homeostasis. Details regarding the nature of these processes, as well as their relationships to each other and to the definitive process of T cell receptor (TCR) gene recombination, are presently emerging. Two widely held concepts are that cell cycle status is inherently and inversely linked to gene recombination and that the outcomes of gene recombination regulate developmental progression. In this study, we analyze TCR-beta recombination and cell cycle status with respect to differentiation during early T cell ontogeny. We find that although differentiation, cell cycle fluctuations, and gene recombination are coincident during normal T cell development, differentiation and cell cycle status are not inherently linked to the recombination process or its products. Rather, recombination appears to occur in parallel with these events as part of a genetically patterned program of development. We propose that the outcome of gene recombination (i.e., TCR expression) may not influence developmental progression per se, but instead serves to perpetuate those developing cells that have been successful in recombination. The potential consequences of this model for the regulation of thymic lymphopoiesis and programmed cell death are discussed.


The origin and purpose of layers of subcutaneous adipose tissue in pigs and man.
Adipose tissue exists in many locations or depots that differ from one another based on numerous and various characteristics. The unique "layered" anatomical feature of subcutaneous adipose tissue (SAT) in man and the pig is reviewed and discussed. The origin of fetal pig adipose tissue subcutaneous layers is reviewed before the onset of adipogenesis and after the overt adipogenesis. Furthermore, the distinguishing characteristics of developing outer SAT layer (OSQ) and middle SAT layer (MSQ) in pigs are reviewed. These characteristics include adipocyte hypertrophy, metabolism and genetic regulation. The MSQ layer is the major layer in the pig and expands to the greatest degree in obesity and growth. Abdominal SAT in man is composed of deep SAT (dSAT) and superficial SAT (sSAT) layers. Clearly, dSAT expands disproportionally more than sSAT with increasing obesity in Caucasian males which precipitates a number of human pathologies associated with increased adiposity. We reviewed the considerable evidence that demonstrates the distinction between sSAT and dSAT which includes higher levels of saturated fatty acids (FAs) and greater levels of lipolysis in dSAT. Furthermore, dSAT expresses more metabolic and inflammatory genes. Studies comparing visceral adipose tissue (VAT) and dSAT indicate that both depots are implicated in insulin resistance (IR) and other human pathologies. Epigenetic studies of MSQ and dSAT have begun to indicate a role for DNA methylation in gene regulation of these depots. Further studies of dSAT and MSQ are warranted as they are clearly a major manifestation of obesity.


Insight and psychosis: Functional and anatomical brain connectivity and self-reflection in Schizophrenia.
Impaired insight into illness, associated with worse treatment outcome, is common in schizophrenia. Insight has been related to the self-reflective processing, centred on the medial frontal cortex. We hypothesized that anatomical and functional routes to and from the ventromedial prefrontal cortex (vmPFC) would differ in patients according to their degree of impaired insight. Forty-five schizophrenia patients and 19 healthy subjects performed a self-reflection task during fMRI, and underwent diffusion tensor imaging. Using dynamic causal modelling we observed increased effective connectivity from the posterior cingulate cortex (PCC), inferior parietal lobule (IPL), and dorsal mPFC (dmPFC) towards the vmPFC with poorer insight and decrease from vmPFC to the IPL. Stronger connectivity from the PCC to vmPFC during judgment of traits related to self was associated with poorer insight. We found small-scale significant changes in white matter integrity associated with clinical insight. Self-reflection may be influenced by synaptic changes that lead to the observed alterations in functional connectivity accompanied by the small-scale but measurable alterations in anatomical connections. Our findings may point to a neural compensatory response to an impairment of connectivity between self-processing regions. Similarly, the observed hyper-connectivity might be a primary deficit linked to inefficiency in the component cognitive processes that lead to impaired insight. We suggest that the stronger cognitive demands placed on patients with poor insight is reflected in increased effective connectivity during the task in this study.


Solidification Behavior of Dy-Tb-Fe Alloys through Experimental Study and Thermodynamic Calculation.
In this work, the solidification microstructure and phase transitions of Dy-Tb-Fe alloy samples were studied by using scanning electron microscopy with energy dispersive spectroscopy (SEM-EDS), X-ray diffraction (XRD) and differential thermal analysis (DTA). No stable ternary compound was detected in the present experiments. The phase transformation temperatures of eight Dy-Tb-Fe alloy samples were measured. Based on the experimental results determined in this work and reported in the literature, the phase equilibria of the Dy-Tb-Fe system was calculated using the CALPHAD method. The calculated vertical sections are consistent with the experimental results determined in this work and reported in the literature. Furthermore, in combination with the experimental solidification microstructure, the solidification behavior of Dy-Tb-Fe alloy samples was analyzed through the thermodynamic calculation with the Gulliver-Scheil non-equilibrium model. The simulated results agree well with the experimental results. This indicates that the reasonable thermodynamic parameters of the Dy-Tb-Fe system were finally obtained.


Measurement of Dating Aggression During Middle School: Structure, Measurement Invariance, and Distinction From General Aggression.
Two studies examined the factor structure of a modified version of the Safe Dates dating aggression scale and evaluated whether dating aggression is distinct from general aggression during early adolescence. Analyses were conducted on a derivation sample of 3,894 adolescents from 37 schools (Study 1) and an independent cross-validation sample of 938 middle school youth (Study 2). Categorical confirmatory factor analyses supported a two-factor model (perpetration and victimization) over models differentiating psychological and physical forms of aggression. The model was invariant across time, sex, grade, and season. Study 2 also supported dating aggression as distinct from general aggression. Results supported measuring dating aggression perpetration and victimization as latent constructs represented by ordered categorical indicators that capture item severity and frequency.


Acetone hydration in supercritical water: (13)C-NMR spectroscopy and Monte Carlo simulation.
The (13)C-NMR chemical shift of acetone delta((13)C[Double Bond]O) was measured in aqueous solution at high temperatures up to 400 degrees C and water densities of 0.10-0.60 g/cm(3) for the study of hydration structure in the supercritical conditions. The average number N(HB) of hydrogen bonds (HBs) between an acetone and solvent waters and the energy change DeltaE upon the HB formation were evaluated from the delta and its temperature dependence, respectively. At 400 degrees C, N(HB) is an increasing function of the water density, the increase being slower at higher water densities. The acetone-water HB formation is exothermic in supercritical water with larger negative DeltaE at lower water densities (-3.3 kcal/mol at 0.10 g/cm(3) and -0.3 kcal/mol at 0.60 g/cm(3)), in contrast to the positive DeltaE in ambient water (+0.078 kcal/mol at 4 degrees C). The corresponding Monte Carlo simulations were performed to calculate the radial and orientational distribution functions of waters around the acetone molecule. The density dependence of N(HB) calculated at 400 degrees C is in a qualitative agreement with the experimental results. In the supercritical conditions, the HB angle in a neighboring acetone-water pair is weakly influenced by the water density, because of the absence of collective HB structure. This is in sharp contrast to the hydration structure in ambient water, where the acetone-water HB formation is orientationally disturbed by the tetrahedral HB network formation among the surrounding waters.


Assessment of the concentration of selected metalloproteinases (MMP-2, MMP-3, MMP-9 and MMP-13) in patients with ulcers as a complication of type 2 diabetes.
Impaired chronic wound healing is a great challenge for modern medicine. This process causes ulceration especially in the course of diseases such as type II diabetes mellitus. This study assesses the concentration of selected matrix metalloproteinases in the example of metalloproteinase 2, 3, 9, 13 in patients with impaired healing of chronic wounds as a complication of type 2 diabetes. Nineteen people took part in the assessment of wound healing in patients with type 2 diabetes. The control group consisted of 21 healthy people. In the blood serum the concentration of MMP-2, MMP-3, MMP-9 and MMP-13 was determined. The concentrations of MMP-2 and MMP-3 in the group of patients with ulcers were significantly higher (61% and 84% accordingly) compared to those in the control group without chronic wounds. No statistically significant differences in MMP-9 and MMP-13 concentrations were observed between the study and control groups. The increase in MMP-2 concentration, which is particularly active in the degradation of type IV collagen, which is the main component of the basal membranes, in patients with type 2 diabetes may impede and delay the healing of chronic wounds and thus contribute to the intensification of vascular complications. In turn, the increase in MMP-3 concentration, which plays a significant role in vascular diseases, in patients with type 2 diabetes may lead to intensification of atherosclerotic changes involving the arteries of the lower extremities and ulceration.


Can a pure motivational interviewing intervention be manualized and still efficacious? A test of feasibility and initial efficacy.
The importance of attending to the therapeutic process despite the challenges in manualizing it is demonstrated in the empirical evolution of motivational interviewing (MI). Whereas manuals exist for adaptations of MI, no manual has been developed and tested for MI in its pure form (pure MI). This study evaluated the feasibility and initial efficacy of a pure MI intervention manual - MI for risky social drinking (MI-RSD) - designed to target risky social drinking behaviors in college students with social anxiety. A pilot sample of 42 college students completed measures of alcohol use and mental health symptoms and the MI-RSD intervention. We developed a manual for the 2-session MI-RSD intervention, trained 4 clinical doctoral students, and used observer-, therapist- and participant-completed measures to evaluate fidelity. Therapists met beginner proficiency in MI fidelity and participant gave high ratings of therapist adherence to MI and working alliance, demonstrating intervention feasibility. Also, participants reported significant reductions in hazardous drinking and evaluation fears, but not in social interaction anxiety. We offer preliminary evidence that pure MI can be manualized and effective. Specifically, MI-RSD represents an alternative to MI adaptations in mitigating alcohol-related harm for young adults. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


A comparison of cetirizine and terfenadine in the management of solar urticaria.
Many solar urticaria patients may benefit from the use of antihistamines. Historically, the value of such therapy was limited by sedation. Newer agents such as terfenadine and cetirizine that are relatively non-sedating appear to be better tolerated by patients. The latter drug, in addition to its antihistamine effect, also appears to inhibit eosinophil migration, which terfenadine and other potent H1 antagonists do not significantly affect. Eosinophils have been reported as early migrating cells in induced solar urticaria, raising the possibility that the dual action of cetirizine may provide a greater potential benefit in the management of solar urticaria. Six patients with idiopathic solar urticaria were entered into a double-blind, phototest study to compare cetirizine and terfenadine. Using the minimal urticarial dose as a phototest end-point, both drugs were equally effective in raising the threshold of sensitivity in 4 patients. Two patients failed to respond to either therapy, which is in keeping with the known variable response to histamine blockade in solar urticaria. At the dosage used, cetirizine therapy appears to be no more effective than terfenadine.


Amplification of over 100 kbp DNA from Single Template Molecules in Femtoliter Droplets.
Reconstitution of the DNA amplification system in microcompartments is the primary step toward artificial cell construction through a bottom-up approach. However, amplification of >100 kbp DNA in micrometer-sized reactors has not yet been achieved. Here, implementing a fully reconstituted replisome of Escherichia coli in micrometer-sized water-in-oil droplets, we developed the in-droplet replication cycle reaction (RCR) system. For a 16 kbp template DNA, the in-droplet RCR system yielded positive RCR signals with a high success rate (82%) for the amplification from single molecule template DNA. The success rate for a 208 kbp template DNA was evidently lower (23%). This study establishes a platform for genome-sized DNA amplification from a single copy of template DNA with the potential to build more complex artificial cell systems comprising a large number of genes.


Antipsychotic drug use complicates assessment of gene expression changes associated with schizophrenia.
Recent postmortem transcriptomic studies of schizophrenia (SCZ) have shown hundreds of differentially expressed genes. However, the extent to which these gene expression changes reflect antipsychotic drug (APD) exposure remains uncertain. We compared differential gene expression in the prefrontal cortex of SCZ patients who tested positive for APDs at the time of death with SCZ patients who did not. APD exposure was associated with numerous changes in the brain transcriptome, especially among SCZ patients on atypical APDs. Brain transcriptome data from macaques chronically treated with APDs showed that APDs affect the expression of many functionally relevant genes, some of which show expression changes in the same directions as those observed in SCZ. Co-expression modules enriched for synaptic function showed convergent patterns between SCZ and some of the APD effects, while those associated with inflammation and glucose metabolism exhibited predominantly divergent patterns between SCZ and APD effects. In contrast, major cell-type shifts inferred in SCZ were primarily unaffected by APD use. These results show that APDs may confound SCZ-associated gene expression changes in postmortem brain tissue. Disentangling these effects will help identify causal genes and improve our neurobiological understanding of SCZ.


Security-Based Passivity Analysis of Markov Jump Systems via Asynchronous Triggering Control.
This article considers the security-based passivity problem for a class of discrete-time Markov jump systems in the presence of deception attacks, where the deception attacks aim to change the transmitted signal. Considering the impact of deception attacks on network disruption, it causes the existence of time-varying delays in signal transmission inevitably, which makes the controlled system and the controller work asynchronously. The asynchronous control method is employed to overcome the nonsynchronous phenomenon between the system mode and controller mode. On the other hand, to reduce the frequency of data transmission, a resilient asynchronous event-triggered control scheme taking deception attacks into account is designed to save communication resources, and the proposed controller can cover some existing ones as special examples. Moreover, different triggering conditions corresponding to different jumping modes are developed to decide whether state signals should be transferred. A new stability criterion is derived to ensure the passivity of the resultant system although there exist deception attacks. Finally, a simulation example is given to verify the theoretical analysis.


Incidence of Epilepsy after Neonatal Seizures: A Population-Based Study.
The incidence of childhood and adolescence epilepsy varies in different areas and over time. Published reports in the Italian pediatric population are few and there is no information on the incidence of epilepsy using the new clinical definition of the disease signed by the International League Against Epilepsy. An increased risk of epilepsy is reported in subjects who presented with neonatal seizures (NS), but few population-based studies are available that compare incidence and age at onset of epilepsy in children with and without NS. Ascertain the incidence of epilepsy in children in the province of Parma by applying the new practical clinical definition of epilepsy, and compare incidence and age of epilepsy onset in children with and without previous NS. The study was carried out in the province of Parma, Emilia-Romagna Region, Northern Italy, using different data sources (clinical records and administrative data), and considered all the children born in the province of Parma between January 2002 and December 2014 and developing epilepsy by December 2016. We calculated the incidence of epilepsy in patients up to 14 years of age, incidence of epilepsy after NS and cumulative incidence of epilepsy at 1, 5, and 10 years' follow-up. To evaluate age at onset of epilepsy, we divided patients into 3 groups (epilepsy onset within 1 month, between 1 and 12 months, and after 1 year of life) and we compared age at onset of epilepsy between patients who had had previous NS and those who had not. The incidence of epilepsy was 78.6/100,000 persons-years (boys 88.1/100,000, girls 68.6/100,000). The incidence of epilepsy after NS was 15.2% (8.2% for male, 23.5% for female; 16.3% in born at term, 14.3% in pre-term). The incidence of epilepsy at 1, 5, and 10 years' follow-up was higher in patients with previous NS than in others. The age at onset of epilepsy was significantly different in the 2 groups, and was younger in those with history of NS: mean age at onset was 10.5 months in those with NS and of 61.8 months in the others. The incidence rate of epilepsy in the Parma district was higher than that reported in other Italian areas studied, probably due to the different methodology used and the application of the most recent definition of epilepsy. Children with NS were at higher risk of epilepsy and develop the disease at a younger age.


Mechanisms of product feedback regulation and drug resistance in cytidine triphosphate synthetases from the structure of a CTP-inhibited complex.
Cytidine triphosphate synthetases (CTPSs) synthesize CTP and regulate its intracellular concentration through direct interactions with the four ribonucleotide triphosphates. In particular, CTP product is a feedback inhibitor that competes with UTP substrate. Selected CTPS mutations that impart resistance to pyrimidine antimetabolite inhibitors also relieve CTP inhibition and cause a dramatic increase in intracellular CTP concentration, indicating that the drugs act by binding to the CTP inhibitory site. Resistance mutations map to a pocket that, although adjacent, does not coincide with the expected UTP binding site in apo Escherichia coli CTPS [EcCTPS; Endrizzi, J. A., et al. (2004) Biochemistry 43, 6447-6463], suggesting allosteric rather than competitive inhibition. Here, bound CTP and ADP were visualized in catalytically active EcCTPS crystals soaked in either ATP and UTP substrates or ADP and CTP products. The CTP cytosine ring resides in the pocket predicted by the resistance mutations, while the triphosphate moiety overlaps the putative UTP triphosphate binding site, explaining how CTP competes with UTP while CTP resistance mutations are acquired without loss of catalytic efficiency. Extensive complementarity and interaction networks at the interfacial binding sites provide the high specificity for pyrimidine triphosphates and mediate nucleotide-dependent tetramer formation. Overall, these results depict a novel product inhibition strategy in which shared substrate and product moieties bind to a single subsite while specificity is conferred by separate subsites. This arrangement allows for independent adaptation of UTP and CTP binding affinities while efficiently utilizing the enzyme surface.


Flexible Transparent Electronic Gas Sensors.
Flexible and transparent electronic gas sensors capable of real-time, sensitive, and selective analysis at room-temperature, have gained immense popularity in recent years for their potential to be integrated into various smart wearable electronics and display devices. Here, recent advances in flexible transparent sensors constructed from semiconducting oxides, carbon materials, conducting polymers, and their nanocomposites are presented. The sensing material selection, sensor device construction, and sensing mechanism of flexible transparent sensors are discussed in detail. The critical challenges and future development associated with flexible and transparent electronic gas sensors are presented. Smart wearable gas sensors are believed to have great potential in environmental monitoring and noninvasive health monitoring based on disease biomarkers in exhaled gas.


A severe complication of anti-TNF alfa treatment.
The antitumor necrosis factor (TNF-alpha) drugs are increasingly used in treating skin diseases such as psoriasis. TNF-alpha is a proinflammatory cytokine with a key role in the pathogenesis of psoriasis but also in host defence against bacterial pathogens, especially against those that multiply inside host cells. The effectiveness of anti-TNF-alpha in the treatment of psoriasis is now widely recognized and has led to their increasingly wide use. Although these drugs are considered relatively safe, their use is associated with an increased incidence of serious infections even in patients treated. Have been described above numerous cases of tuberculosis but has also observed an increased incidence of granulomatous infections by intracellular bacteria such as Legionella pneumophila required. Infections due to this biotic agent, if not diagnosed early, are potentially fatal. We report the case of a patient, heavy smoker, suffering from severe skin psoriasis who after starting treatment with infliximab developed a pneumonia caused by Legionella pneumophila. Our aim is to draw the attention of specialists on increasing risk of granulomatous infections by intracellular agents in patients being treated with anti TNF-alpha.


Molecular mechanism of hypoxia-induced chondrogenesis and its application in in vivo cartilage tissue engineering.
Cartilage engineering is one of the most challenging issue in regenerative medicine, due to its limited self-ability to repair. Here, we assessed engineering of cartilage tissue starting from human bone marrow (hBM) stem cells under hypoxic environment and delineated the mechanism whereby chondrogenesis could be conducted without addition of exogenous growth factors. hBM stem cells were cultured in alginate beads and chondrogenesis was monitored by chondrocyte phenotypic markers. Activities and roles of Sox and HIF-1α transcription factors were investigated with complementary approaches of gain and loss of function and provided evidences that HIF-1α is essential for hypoxic induction of chondrogenesis. Thereafter, hBM cells and human articular chondrocytes (HAC) underwent chondrogenesis by 3D and hypoxic culture for 7 days or by ectopic expression of HIF-1α. After subcutaneous implantation of 3 weeks into athymic mice, tissue analysis showed that hypoxia or HIF-1α overexpression is effective and sufficient to induce chondrocyte phenotype in hBM cells, without use of exogenous growth factors. Therefore, this study brings interesting data for a simple and affordable system in biotechnology of cartilage engineering.


Synthesis and Characterization of Dye-Doped Au@SiO2 Core-Shell Nanoparticles for Super-Resolution Fluorescence Microscopy.
Dye-doped nanoparticles have been investigated as bright, fluorescent probes for localization-based super-resolution microscopy. Nanoparticle size is important in super-resolution microscopy to get an accurate size of the object of interest from image analysis. Due to their self-blinking behavior and metal-enhanced fluorescence (MEF), Ag@SiO2 and Au@Ag@SiO2 nanoparticles have shown promise as probes for localization-based super-resolution microscopy. Here, several noble metal-based dye-doped core-shell nanoparticles have been investigated as self-blinking nanomaterial probes. It was observed that both the gold- and silver-plated nanoparticle cores exhibit weak luminescence under certain conditions due to the surface plasmon resonance bands produced by each metal, and the gold cores exhibit blinking behavior which enhances the blinking and fluorescence of the dye-doped nanoparticle. However, the silver-plated nanoparticle cores, while weakly luminescent, did not exhibit any blinking; the dye-doped nanoparticle exhibited the same behavior as the core fluorescent, but did not blink. Because of the blinking behavior, stochastic optical reconstruction microscopy (STORM) super-resolution analysis was able to be performed with performed on the gold core nanoparticles. A preliminary study on the use of these nanoparticles for localization-based super-resolution showed that these nanoparticles are suitable for use in STORM super resolution. Resolution enhancement was two times better than the diffraction limited images, with core sizes reduced to 15 nm using the hybrid Au-Ag cores.


Cardiac implantable electronic device remote monitoring in a large cohort of patients and the need for planning.
Aim: The remote monitoring (RM) of cardiac implantable electronic devices (CIED) is standard of care. We describe an organizational and projection RM workload model. Methods: At the time of the analysis (2015), 3995 CIED patients were followed-up; 1582 (40.5%) with RM. All RM transmissions (Tx) have been gathered in five event types. Results: We received 10,406 Tx, classified as: 128 (1.2%) red alerts, 141 (1.3%) atrial fibrillation episodes, 1944 (18.6%) yellow alerts, 403 (3.9%) lost Tx (disconnected/noncompliant patients) and 7790 (75.0%) Tx 'OK' (un-eventful Tx). At the time of 100% of remote CIED managed, we can expect a total of 25,990 Tx/year. Conclusion: We provide a descriptive analysis of remote monitoring management and workload estimation in a large cohort of CIED patients.


[Chemical denitrification of nitrate from groundwater].
Batch experiments for catalytic reduction of nitrate from groundwater with Pd and/Cu catalysts were conducted. It was found that Pd-Cu combined catalysts at a ratio of 4 can maximize the nitrate reduction into nitrogen; above 80% total nitrogen removal efficiency was realized in this study. It was also found that the catalytic activity was affected by the amounts of catalysts, hydrogen flow rate and pressure, the initial concentration of nitrate. With an increasing in the amount of the catalysts, both nitrite and ammonia intermediates can be kept at a low level. A high flow rate and pressure of hydrogen was in favor of the catalytic reduction of nitrate, but not benefit for the selectivity. The catalytic reduction followed a first order kinetics in term of the initial nitrate concentration.


Villous adenoma: a rare tumor of vaginal vault.
Villous adenomas are extremely rare tumors in the vagina and are indistinguishable from their colonic counterparts. These are considered as enteric type neoplasms of lower female genital tract arising from cloacal remnants. The significance of this lesion lies in the fact that it has to be distinguished from metastatic adenocarcinoma from other sites. We present a case of villous adenoma of vaginal vault, a rare site of presentation in a 30-year-old female.


Global effects of histone modifications.
The relationship between chromatin structure and transcriptional regulation has been the focus of many research publications over the past few years. A lot of progress has been made in understanding the role of histone modifications. The interdependence of histone methylation and DNA methylation has been revealed. Methylation, acetylation, phosphorylation and ubiquitination of specific histone residues are all able to affect the DNA binding of transcription factors and to change the structure of chromatin on a genome-wide scale. This review aims to summarise some of the experiments that demonstrated the global effects of the modifications of the five histone proteins, H1, H2A, H2B, H3 and H4.


Asymptotic distributions of periodically driven stochastic systems.
We study the large-time behavior of Brownian particles moving through a viscous medium in a confined potential, and which are further subjected to position-dependent driving forces that are periodic in time. We focus on the case where these driving forces are rapidly oscillating with an amplitude that is not necessarily small. We develop a perturbative method for the high-frequency regime to find the large-time behavior of periodically driven stochastic systems. The asymptotic distribution of Brownian particles is then determined to second order. To first order, these particles are found to execute small-amplitude oscillations around an effective static potential that can have interesting forms.


Tolerance to baked and fermented cow's milk in children with IgE-mediated and non-IgE-mediated cow's milk allergy in patients under two years of age.
IgE-mediated cow's milk allergy (CMA) has been shown consistent in milder heated-milk tolerant and severe heated-milk reactant groups in patients older than two years. Little is known whether fermentation of milk gives rise to similar clinical phenotypes. We aimed to determine the influence of extensively heated and fermented cow's milk on the IgE-mediated and non-IgE-mediated CMA in children younger than two years. Subjects followed with the diagnosis of IgE-mediated and non-IgE-mediated CMA for at least six months underwent unheated milk challenge. IgE-mediated and non-IgE-mediated groups were categorised as unheated milk-reactive and tolerant, separately. Unheated milk-reactive groups were further challenged sequentially with fermented milk (yoghurt) and baked milk, 15 days apart. Allergy evaluation with skin tests, prick-to-prick tests and atopy patch tests were performed. Fifty-seven children (median age: 14 months; range: 7-24 months) underwent unheated milk challenge. Eleven of 27 children with IgE-mediated CMA and 14 of 30 children with non-IgE-mediated CMA tolerated unheated milk. Among subjects who reacted to unheated milk; 15 of 16 subjects (93%) with IgE-mediated CMA also reacted to yoghurt, whereas 11 of 16 subjects (68%) with non-IgE-mediated CMA tolerated fermented milk. Thirteen subjects (81%) of the unheated milk-reactive IgE-mediated group tolerated to heated milk. None of 16 subjects of unheated milk-reactive non-IgE-mediated group reacted to baked milk. The majority of children under the age of two years with both IgE-mediated and non-IgE-mediated CMA tolerated baked-milk products. Yoghurt was tolerated in two thirds of unheated milk reactive patients suffering from non-IgE-mediated CMA.


Pre-and postoperative Tc 99m scanning in monitoring pulmonary perfusion in the tetralogy of Fallot.
Angiocardiopneumographic scanning with Tc 99m was used in pulmonary perfusion and intracardiac shunt before and after radical corrective surgery of the tetralogy of Fallot. The study was carried out on 25 patients with the tetralogy of Fallot undergoing radical treatment with infundibular patch. Preoperative scanning has shown increased thickening of right ventricle and contemporary injection of pulmonary artery and aorta, the latter with a much higher radioactivity; the obstruction to the right flow was not clearly defined with respect to morphology and function, pulmonary perfusion has shown an extremely poor radioactivity and prolonged duration. Scanning following surgical treatment showed normal right and left phase and absence of residual shunts. Captation of marker in the lungs was always complete. Such data illustrate the many advantages gained by the use of non invasive diagnostic procedures in cardiology and "the first pass technique" in the evaluation of results of surgical treatment.


Multiplex minisequencing of the 21-hydroxylase gene as a rapid strategy to confirm congenital adrenal hyperplasia.
Congenital adrenal hyperplasia (CAH) is a frequent autosomal recessive disease, with a wide range of clinical manifestations, most commonly attributable to mutations in the 21-hydroxylase gene (CYP21). Large gene deletions, large gene conversions, a small 8-basepair deletion, and eight point mutations in CYP21 account for approximately 95% of all enzyme deficiencies. We developed a new strategy for a rapid CYP21 analysis. DNA samples from 40 CAH patients previously genotyped by direct DNA sequencing were reanalyzed by allele-specific amplification of the functional CYP21 gene followed by a multiplex minisequencing reaction using 13 primers. In addition, a second PCR that amplified a part of exon 3 was used to demonstrate the presence or absence of at least one functional gene. The assay detected the P453S mutation and nine of the most common mutations (P30L, intron 2 splice, Delta 8bp, I172N, exon 6 cluster, V281L, F306+t, Q318X, and R356W) caused by microconversions from the CYP21P pseudogene. The concordance was 100% for detecting these mutations, including gene deletions and large gene conversions. The 40 patient DNA samples were analyzed in 1.5 working days by one technician (actual hands-on time, 3.5 h). The material cost for analyzing one sample was approximately 10.00 Euros (US $9.00). This novel mutation screening strategy rapidly detects 90-95% of all mutations associated with CAH and appears applicable as a tool for confirmation of increased 17-hydroxyprogesterone found in neonatal CAH screening.


[Research advance of ANRIL on atherosclerosis by regulating cell proliferation and apoptosis].
Atherosclerosis is an important pathological basis for coronary artery disease. ANRIL is an antisense non-coding RNA located in Chr9p21 locus, which was identified as the most significant risk locus associated with atherosclerosis. ANRIL can produce multiple transcripts including linear and circular transcripts after various transcript splicing. It has been illustrated that ANRIL plays important roles in the pathology of atherosclerosis by regulating the proliferation and apoptosis of vascular cells. Linear ANRIL can regulate the proliferation of vascular smooth muscle cells (VSMCs) in plaques by chromatin modification, as well as influence the proliferation and the apoptosis of macrophages in post transcription; circular ANRIL can affect the proliferation and apoptosis of VSMCs by chromatin modification as well as interfering with rRNA maturation. In this review, we describe the ANRIL evolution, different transcripts characteristics, and their roles in the proliferation and apoptosis of vascular cells to participate in the process of atherosclerosis, for further understanding the pathogenesis of atherosclerosis and finding potential targets for diagnosis and treatment of atherosclerosis.


Fibrous inflammatory pseudotumor of the bladder.
Fibrous pseudotumor of the bladder, a rare, benign, and proliferative lesion of the submucosal stroma, can be mistaken on gross examination for a malignant lesion and must be differentiated on histologic examination from several bladder malignancies. Radiographic examination alone cannot establish a definitive diagnosis. Complete transurethral resection of such lesions appears to be curative.


Functional characterization of naturally occurring wild soybean mutant (sg-5) lacking astringent saponins using whole genome sequencing approach.
Triterpenoid saponins are one of the most highly accumulated groups of functional components in soybean (Glycine max) and the oxidative reactions during their biosynthesis are required for their aglycone diversity. Natural mutants of soyasaponins in wild soybean (Glycine soja) are valuable resources for establishing the soyasaponin biosynthesis pathway and breeding new soybean varieties. In this study, we investigated the genetic mechanism behind the absence of group A saponins in a Korean wild soybean mutant, CWS5095. Whole genome sequencing (WGS) of CWS5095 identified four point mutations [Val6 → Asp, Ile231 → Thr, His294 → Gln, and Arg376 → Lys] in CYP72A69 (Glyma15g39090), which oxygenate the C-21 position of soyasapogenol B or other intermediates to produce soyasapogenol A, leading to group A saponin production. An in vitro enzyme activity assay of single-sited mutated clones indicated that the Arg376 > Lys mutation (a highly conserved mutation based on a nucleotide change from G → A at the 1,127th position) may lead to loss of gene function in the sg-5 mutant. A very high normalized expression value of 377 reads per kilo base per million (RPKM) of Glyma15g39090 in the hypocotyl axis at the early maturation seed-development stage confirmed their abundant presence in seed hypocotyls. A molecular dynamics analysis of the Arg376 > Lys mutation based on the CYP3A4 (a human CYP450) protein structure found that it was responsible for the increase in axis length toward the heme (active site), which is critically important for biological activity and ligand binding. Our results provide important information on how to eradicate bitter and astringent saponins in soybean by utilizing the reported mutation in Glyma15g39090, and its importance for seed hypocotyl development based on transcript abundance.


Art education lecturers' intention to continue using the blackboard during and after the COVID-19 pandemic: An empirical investigation into the UTAUT and TAM model.
The success of faculty in adopting technology in this digital era has a direct impact on the success of the students and, eventually, the educational institution. Many teachers, on the other hand, have yet to implement technological tools such as Canvas into their classes. As a result, this study looked at art universities lecturers' opinions of variables influencing actual blackboard use, as well as their desire to utilize the lesson plan in learning and teaching during the COVID-19 Pandemic. The TAM model and the unified theory of acceptance and use of technology (UTAUT) were used to analyze lecturers' satisfaction with the blackboard and their desire to continue using it, as well as the actual usage of blackboards. A survey of 159 professors using Canvas in art instruction at five Saudi Arabian public institutions yielded the research findings. In addition, structural equation modeling (SEM) was used to evaluate the study model as well as the mediating relationship between factors influencing the desire to implement the lesson in learning and teaching during the COVID-19 Pandemic. According to the study's findings, superior impact, performance expectation, effort expectations, enabling conditions, and reported enjoyment all had statistically significant effects on perceived usefulness (PU) and perceived ease of use. The current study discovered that PU and perception of use had statistically significant influence on behavior intent, actual blackboard usage, and lecturers' happiness with utilizing blackboard in learning and teaching during the COVID-19 Pandemic. Furthermore, the results demonstrate that lecturer's intention to continue utilizing chalkboard in learning and teaching during and after COVID-19 Pandemic was influenced by actual usage of blackboard and reported pleasure. The findings are useful for education based, regulators, and practitioners who seek to create and enhance effective methods to use e-learning systems during and after the COVID-19 Pandemic.


Youngest Composite Full-Face Transplant: A Model for Vascularized Composite Allograft in Younger Populations.
The field of face transplantation continues to evolve, with more complex defects being addressed, and, at the same time, increased outcome expectations. Given our unique long-term experience in this field, we consented one of the youngest patients to undergo a full-face transplant. An 18-year-old woman presented with complete destruction of her central face and craniofacial structures. She had coexisting major injuries, including pituitary gland, visual axis, and motor control. After extensive rehabilitation and reconstruction techniques, the patient underwent face transplant on May 4, 2017, at the age of 21 years. The total operative time for the recipient was 26 hours. There were no major perioperative complications. Since transplant, the patient has undergone 3 revision surgeries. She is near completely independent from a daily life activity standpoint. She has had 1 episode of rejection above grade II that was successfully treated with a short-term increased in immunosuppression. Contrary to data in solid organ transplantation where youth is associated with increased risk of rejection, our current algorithm in immunosuppression, combined with this patient's compliance, has led to only 1 rejection episode beyond grade II. This successful transplant can serve as a model for future vascularized composite transplants in younger populations.


Use of kinetic isotope effects to delineate the role of phenylalanine 87 in P450(BM-3).
The substrate oxidation rates of P450(BM-3) are unparalleled in the cytochrome P450 (CYP) superfamily of enzymes. Furthermore, the bacterial enzyme, originating from Bacillus megaterium, has been used repeatedly as a model to study the metabolism of mammalian P450s. A specific example is presented where studying P450(BM-3) substrate dynamics can define important enzyme-substrate characteristics, which may be useful in modeling omega-hydroxylation seen in mammalian P450s. In addition, if the reactive species responsible for metabolism can be controlled to produce specific products this enzyme could be a useful biocatalyst. Based on crystal structures and the fact that the P450(BM-3) F87A mutant produces a large isotope in contrast to the native enzyme, we propose that phenylalanine 87 is responsible for hindering substrate access to the active oxygen species for nonnative substrates. Using kinetic isotopes and two aromatic substrates, p-xylene and 4,4'-dimethylbiphenyl, the role phenylalanine 87 plays in active-site dynamics is characterized. The intrinsic KIE is 7.3 +/- 2 for wtP450(BM-3) metabolism of p-xylene. In addition, stoichiometry differences were measured with the native and mutant enzyme and 4,4'-dimethylbiphenyl. The results show a more highly coupled substrate/NADPH ratio in the mutant than in the wtP450(BM-3).


Exercise testing in occupational lung diseases.
The authors discuss the value of exercise testing in two areas of importance in occupational pulmonary disease: (1) the surveillance of worker populations in order to gain epidemiologic and physiologic understanding of disease and (2) the assessment of work capacity in individual patients.


Histochemical identification of human T lymphocytes from paraffin sections.
The alpha-naphthyl acetate esterase (ANEA) is a histochemical marker for human T lymphocytes in cell smears and frozen tissue sections. We have now applied the ANAE method to paraffin-embedded tissue sections. We first demonstrated with cytocentrifuged cell smears of blood leukocytes that the ANAE activity is preserved upon prolonged storage in formol calcium, Holt's buffer, acetone, xylene, and heat. When the tissue sections were similarly processed and embedded in paraffin, the ANAE positive (T) lymphocytes were identified by their distinct display of one or more reddish-brown reaction dots in the cell cytoplasm. ANAE positive mononuclear phagocytes were easioy distinguished from the T lymphocytes by their diffuse, sodium fluoride-sensitive pancytoplasmic reaction. The extension of the ANAE method to paraffin-embedded tissue sections with superior morphological integrity, makes it possible to apply it in practical biopsy pathology.


Unboxing cluster heatmaps.
Cluster heatmaps are commonly used in biology and related fields to reveal hierarchical clusters in data matrices. This visualization technique has high data density and reveal clusters better than unordered heatmaps alone. However, cluster heatmaps have known issues making them both time consuming to use and prone to error. We hypothesize that visualization techniques without the rigid grid constraint of cluster heatmaps will perform better at clustering-related tasks. We developed an approach to "unbox" the heatmap values and embed them directly in the hierarchical clustering results, allowing us to use standard hierarchical visualization techniques as alternatives to cluster heatmaps. We then tested our hypothesis by conducting a survey of 45 practitioners to determine how cluster heatmaps are used, prototyping alternatives to cluster heatmaps using pair analytics with a computational biologist, and evaluating those alternatives with hour-long interviews of 5 practitioners and an Amazon Mechanical Turk user study with approximately 200 participants. We found statistically significant performance differences for most clustering-related tasks, and in the number of perceived visual clusters. Visit git.io/vw0t3 for our results. The optimal technique varied by task. However, gapmaps were preferred by the interviewed practitioners and outperformed or performed as well as cluster heatmaps for clustering-related tasks. Gapmaps are similar to cluster heatmaps, but relax the heatmap grid constraints by introducing gaps between rows and/or columns that are not closely clustered. Based on these results, we recommend users adopt gapmaps as an alternative to cluster heatmaps.


The effect of gentamicin on acetylsalicylic acid-induced platelet antiaggregatory action in mouse pial arterioles.
Gentamicin (G) treatment (5, 20, 40 and 80 mg kg[-1] day[-1] given intramuscularly for 6 days) was shown to cause a dose-related platelet proaggregatory effect in mouse pial microcirculation. This was associated with a reduction in mouse renal function, indicated by high plasma creatinine and urea concentrations. When G was given at the same doses but as a single injection, it caused no change in renal function or platelet aggregation. Gentamicin (20 and 80 mg kg/day, given intramuscularly for 6 days) significantly (P < 0.05) impeded the platelet antiaggregatory effect of acetylsalicylic acid (100 mg kg[-1], intraperitoneally).


Potential SARS-COV preclinical (in vivo) compounds targeting COVID-19 main protease: a meta-analysis and molecular docking studies.
Due to the migratory flow of infected people with severe acute respiratory syndrome virus (SARS COV-2), the number of confirmed cases of coronavirus disease 2019 (COVID-19) is accelerating globally; preclinical evidence of antiviral agents that can combat this pandemic is still elusive. We identified published articles on SARS-COV efficacy experiments in which some selected compounds were used to test the reduction of the virus load in mice. A systematic search of articles was conducted in PubMed, Web of Science, and Scopus. We then developed a combined model based on a systematic review, meta-analyses, and molecular docking studies to evaluate the effect size of preclinical studies of compounds that have been tested against SARS-COV. Because substantial heterogeneity was expected, random effect model meta-analyses were carried out to estimate the overall pooled disease's prevalence. All meta-analyses were performed with Stata version 15.0. Subgroup analyses on therapies were conducted as well. Molecular docking studies of the inhibitors in the active pocket of COVID-19 protease were also performed. From all screened articles, six studies were appropriate for ultimate meta-analysis and systematic review. The residual amount of heterogeneity was high (τ2 =0.02; heterogeneity I2 =85.5 % with heterogeneity chi-square =103.57, a degree of freedom =15, and p <0.001). The overall random pooled prevalence of infected mice treated with the selected compounds was 78.1 % [95 % Confidence Interval (CI): 14.7-17.0 %]. Prophylactic has a significantly higher pooled prevalence than therapeutic, with 21.8 % (95 % CI: 16.4 % to 28.8 %). Our results indicated that most of the SARS-COV inhibitors analyzed were less effective in reducing the lung virus titer of SARS-COV infection in animal models. The findings from molecular docking studies also identified COVID-19 inhibitors that are good for optimization and drug development to fight against COVID-19 infection. Findings from the review showed that studies on the preclinical compounds targeting SARS-COV and COVID-19 are limited. Furthermore, molecular docking studies and meta-analysis results substantiated three compounds, i.e., EIDD-2801, GS-5734, and amodiaquine. HIPPOKRATIA 2020, 24(3): 99-106.


Diminished amyloid-β uptake by mouse microglia upon treatment with quantum dots, silver or cerium oxide nanoparticles: Nanoparticles and amyloid-β uptake by microglia.
Alzheimer's disease (AD) is a chronic neurodegenerative disease leading to progressive dementia in elderly people. The disease is characterized, among others, by formation of amyloid-β (Aβ) polypeptide plaques in the brain. Although etiology of the disease is not fully understood, recent research suggest that nanomaterials may affect AD development. Here, we described the consequences of exposure of mouse BV-2 microglia to silver nanoparticles (AgNPs, 50 µg/mL), cerium oxide nanoparticles (CeO2NPs, 100 µg/mL), and cadmium telluride quantum dots (CdTeQDs, 3 or 10 µg/mL) in the context of its ability to clear Aβ plaques. The brain microglial cells play an important role in removing Aβ plaques from the brain. Cell viability and cycle progression were assessed by trypan blue test and propidium iodide binding, respectively. The uptake of Aβ and NPs was measured by flow cytometry. Secretion of proinflammatory cytokines was measured with the use of cytometric bead array. Aβ (0.1 μM) did not affect viability, whereas NPs decreased microglia growth by arresting the cells in G1 phase (CdTeQDs) or in S phase (AgNPs and CeO2NPs) of cell cycle. The uptake of Aβ was significantly reduced in the presence of AgNPs and CeO2NPs. In addition, the least toxic CeO2NPs induced the release of proinflammatory cytokine, tumor necrosis factor α. In summary, each of the NPs tested affected either the microglia phagocytic activity (AgNPs and CeO2NPs) and/or its viability (AgNPs and CdTeQDs) that may favor the occurrence of AD and accelerate its development.