Effect of supercoiling on the abortive initiation kinetics of the RNA-I promoter of ColE1 plasmid DNA.
An analysis of the effect of DNA supercoiling on transcription of the RNA-I promoter of ColE1 has been undertaken using the abortive initiation assay developed by McClure (McClure, W. R., Cech, C. L., and Johnston, D. E. (1978) J. Biol. Chem. 253, 8941-8948). A tau plot was constructed, enabling the equilibrium constant for closed complex formation (KI) and the isomerization rate constant (k2) to be determined. The effect of supercoiling on RNA-I is an apparent 100-fold increase in the isomerization rate, k2, with little or no effect on KI. This is the first reported abortive initiation analysis of RNA-I transcription which serves in the regulation of plasmid replication.


Non-A, non-B hepatitis: clinical laboratory course in patients on hemodialysis and its correlation with the presence of anti-hepatitis C virus antibodies.
136 patients on hemodialysis, 89 males and 47 females, were studied; we evaluated the index of hepatic function (SGOT and SGPT) and antibodies against HCV. We observed 42 cases of increased transaminases classified as non-A, non-B (NANB) hepatitis. Antibodies against HCV were present in 40 patients. Among 42 patients with NANB hepatitis. 31 (73.8%) presented anti-HCV antibodies. No significant clinical or laboratory difference exists between anti-HCV-positive and -negative patients with NANB hepatitis. The distribution of patients who present anti-HCV antibodies is similar in post-transfusional and sporadic forms.


Evidence that cytoplasmic birefringence is a normal feature of rat adrenal chromaffin cells.
The rat adrenal medulla was fixed by isobaric whole-body perfusion with glutaraldehyde. Specimens treated with anisotropic stains exhibited a diffuse cytoplasmic birefringence. Evidence is presented that birefringence is not only associated with "dark" cell formation, but may be a normal feature of fixed chromaffin cells. It is suggested that the birefringence is caused by the fixed contents of the chromaffin vesicles and/or the microtrabecular lattice recently described in rat chromaffin cells.


[Lacrimal systems endoscopy with a new endoscope system].
Successful therapy of postsaccal stenosis in the lacrimal system can be done by an ENT Surgeon by endonasal dacryocystorhinostomy. It is desirable to have a good diagnostic method of the lacrimal system before undertaking surgery. This study describes the first experiences of endoscopic diagnostics of the lacrimal system with a new endoscope. 24 patients (10 male and 14 female, between 3 and 65 years old) with epiphora and chronic lacrimal system infections were examined in the ENT Department Homburg/Saar with a 0.53 mm modular endoscope, between 11/2008 and 12/2009. The result were evaluated and analysed. The endoscopic lacrimal system examination was done in all 24 patients. 8 patients with high stenosis postsaccal and purulent epiphora had an endonasal dacryocystorhinostomie. 13 patients with prasaccal stenosis or stenosis in the ductus nasolacrimalis with clear epiphora were treated with irrigation, dilatation and U-intubation. The removal of the Endoscopic dilatation was undertaken in 3 children without using bicanicular intubation. Lacrimal system endoscopy with a modular endoscope with the possibility of interventional procedures is an efficient option in the widening diagnosis and therapy of the lacrimalm system disease. With this technique is it possible with selected patients to do interventional therapy. On the basis of the endoscopic findings an interventional therapy or an endonasal dacryocystorhinostomie can be done.


[Standardization of the resistance of grounded electrocardiographs with patient circuits ground-insulated to electrical cophase interferences from an external main].
The possibility of simulating conditions for electrocardiographic interferences examination by current and voltage generators is shown. These two models allow it to adopt for appraising the resistance of electrocardiographs to outside cophasic electric interferences any one of the two equivalent parameters, viz. "the equivalent resistance of cophasic interferences" and "coefficient of cophasic rejection". The said parameters represent the relation between the differential value of the interferences voltage at the inlet of the electrocardiograph and the interferences current and interferences voltage of the current generators and of the interverences voltage that produced the former, respectively. The required magnitudes for these parameters have been found, namely less than or equal to 14 Om and 0.5-10(-6) V. For practical purposes it is preferable to use the coefficient of cophasic rejection.


Peptides and monoclonal antibodies which bind to platelet glycoproteins IIb and/or IIIa inhibit clot retraction.
The rate of clot retraction in platelet-rich plasma was decreased by synthetic peptide analogues of fibrinogen and monoclonal antibodies each of which bind to the platelet plasma membrane glycoproteins IIb and/or IIIa. These and related data demonstrate that intact complexes of the glycoproteins IIb and IIIa are required for platelet-mediated clot retraction.


Factors Associated with Vaccine Hesitancy in the State of Alaska.
Vaccine hesitancy (VH) is a major public health problem which includes not only concerns about vaccine, but often includes beliefs not aligned with evidence. The etiology of VH is complicate, including genuine concerns about vaccine safety to beliefs in disproven or unsubstantiated theories. Understanding VH includes determining the factors associated with people most likely to be susceptible. To identify characteristics of persons more likely to have VH based on perceived vaccine safety. We conducted a cross sectional, targeted online survey of 1,024 respondents. The survey collected data on demographics and vaccine beliefs. A seven-question VH scale was created based on an extensive literature review. When measured, this scale had high internal validity based on Cronbach's Alpha (α = 0.80, 95% CI, 0.79, 0.82) and provided a continuous measurement to understand the VH degree. Persons reporting a conservative political ideology had the highest VH score ([Formula: see text] = 15.0), followed by other ([Formula: see text] =12.7), moderate ([Formula: see text] =9.29), and liberal ([Formula: see text] =6.72). Education level was another strong indicator of VH, decreasing from the highest score of high school graduates ( [Formula: see text] = 13.2) to graduate degree holders ([Formula: see text] = 9.22). The more conservative a person's political ideology, the higher the VHS. We asked for ideology, rather than political party, although they are related, they are not the same. Ideology influences more domains than voting patterns. Higher educational attainment shows a protective effect against VH. These findings have direct implication for public health interventions, new interventions should be designed to be more accessible for persons with less formal education and more amenable to persons who hold a more conservative ideology.


Corrigendum to "Characterization of Zebrafish von Willebrand Factor Reveals Conservation of Domain Structure, Multimerization, and Intracellular Storage".
[This corrects the article DOI: 10.1155/2012/214209.].


Assessment of controls layout of Indian tractors.
Tractors in low-income countries are used both for farm and non-farm activities. Most of the tractors being manufactured in India are products of collaboration with other countries. The design of tractors manufactured in India has not changed much in the past five decades especially from an ergonomics point of view, because of economic considerations. This paper describes a tractor control layout assessment with respect to the Indian population and compares the location of controls with workspace envelopes and the IS12343 standard for commonly used tractors on Indian farms. Controls like steering, foot clutch, foot brake, foot accelerator are located in areas defined by IS12343 standard in some tractors but these are not placed in the workspace envelopes of the Indian population. This results in a mismatch between the workspace envelope and location of controls as defined by the standard. The controls need a complete change in their layout to be in the workspace envelopes, as this cannot be achieved by providing seat movement in the horizontal and vertical directions in the present tractor design.


Findings of electroencephalographic brain mapping in mild to moderate dementia of the Alzheimer type during resting, motor, and music-perception conditions.
Sixteen-channel electroencephalographic (EEG) brain mapping studies were carried out under resting conditions in 41 patients with mild to moderate dementia of the Alzheimer type, diagnosed according to DSM-III-R criteria. The patients showed diffuse patterns of EEG slowing, as known from the literature. They showed increases of theta power in large bilateral brain areas and bitemporal increases of delta power as compared with both age-matched and younger control subjects. In addition, the patients had increased beta activity in comparison with the younger control group, but not with the age-matched control subjects. Functional brain mapping revealed decreases of EEG power during manual-motor and music-perception tasks in delta frequencies for patients, which was not seen in either of the two control groups. All frequency bands revealed major gender-related differences in EEG activity, which should receive more attention in future research.


In vitro production of pregnancy-associated plasma protein A by human decidua and trophoblast.
Immunohistochemical techniques and direct measurements of pregnancy-associated plasma protein A (PAPP-A) have demonstrated the presence of PAPP-A in trophoblast and decidua. The purpose of the present study was to investigate the possibility that these tissues are capable of producing PAPP-A in vitro. Trophoblast and decidua were obtained from term deliveries and from legal surgical terminations of pregnancy (7 to 12 weeks). In addition to trophoblast and decidua, myometrium was also obtained during two hysterectomies in the first trimester of pregnancy. Tissues were incubated in medium 199 at 37 degrees C under an oxygen/carbon dioxide atmosphere. Media containing either pregnancy-associated serum or non-pregnancy-associated serum were changed after 8 hours of incubation in medium 199 alone. In addition to PAPP-A, human placental lactogen (hPL) and prolactin (Prl) were measured in homogenates and media by radioimmunoassays in order to confirm the viability of the cultured tissues. Addition of pregnancy-associated serum to the media induced a significant release of PAPP-A from trophoblast and decidua when compared to that in control cultures. Non-pregnancy-associated serum had no effect. Myometrium did not release any measurable PAPP-A into the medium even in the presence of pregnancy-associated serum. Cycloheximide added to pregnancy-associated serum significantly inhibited the release of PAPP-A from trophoblast and decidua. These last tissues, irrespective of the culture condition, released significantly more PAPP-A as well as hPL and Prl than was initially present in the tissue. These data demonstrate that PAPP-A is released in vitro by trophoblast and decidua (but not by myometrium) and that this release can be magnified by a factor present only in pregnancy-associated serum. The release of PAPP-A, hPL, and Prl is considered as a de novo production since concentration of these proteins are higher in media and tissues after incubation compared to concentrations initially present in the tissue before culture and since cycloheximide significantly inhibits the release of PAPP-A, Prl, and hPL from the cultured tissues.


The influence of vitamin D metabolites on the calcification of cartilage matrix and the C-propeptide of type II collagen (chondrocalcin).
The influence of vitamin D metabolites (at 1 X 10(-10) M) on the calcification of cartilage matrix (measured by 45Ca2+ uptake) and the C-propeptide of type II collagen (measured by radioimmunoassay) has been studied using organ cultures and chondrocytes isolated from growth plates of vitamin D-deficient and -sufficient 11-day-old rats. Vitamin D-deficient rats had reduced amounts of C-propeptide in their serum and freshly isolated growth plate chondrocytes. In all chondrocytes cultured from vitamin D-deficient animals, the C-propeptide content was maximal at 24 hr whereas calcification continued to increase for up to 72 hr. In organ and chondrocyte cultures of tissue from vitamin D-sufficient rats, both 1,25-dihydroxycholecalciferol (1,25(OH)2D3) and 24,25-dihydroxycholecalciferol (24,25(OH)2D3) were required for maximal stimulation of calcification and maximal increases in C-propeptide content. In these D-replete tissues, 24,25-(OH)2D3 had a less stimulatory effect on both calcification (organ and cell cultures) and C-propeptide (organ cultures only), while 1,25(OH)2D3 alone had no effect in cell cultures but an inhibitory effect in organ cultures. Studies of cells or tissue from growth plates of vitamin D-deficient rats demonstrated that 24,25(OH)2D3 alone produced maximal calcification and maximal increases in the C-propeptide content. 1,25(OH)2D3 generally had an inhibitory effect on both calcification and C-propeptide when used alone. In the presence of 1,25(OH)2D3, the stimulatory effect of 24,25(OH)2D3 was partly abrogated. Maximal stimulation of calcification and increases in C-propeptide by 24,25(OH)2D3 were observed at 1 X 10(-9) M and 1 X 10(-10) M. In none of these studies was there any effect on proteoglycan content.(ABSTRACT TRUNCATED AT 250 WORDS)


Prenatal diagnosis and molecular cytogenetic characterization of a de novo 3.19-Mb chromosome 14q32.13-q32.2 deletion of paternal origin.
We present prenatal diagnosis and molecular cytogenetic characterization of a de novo 3.19-Mb chromosome 14q32.13-q32.2 deletion of paternal origin. A 36-year-old woman underwent amniocentesis at 20 weeks of gestation because of an advanced maternal age. Her husband was 36 years old. Amniocentesis revealed a karyotype of 46,XY,del(14)(q32.1q32.2). Simultaneous array comparative genomic hybridization (aCGH) analysis showed the result of a 14q32.13-q32.2 deletion. Prenatal ultrasound was unremarkable. The parental karyotypes were normal and did not have such a deletion. The pregnancy was subsequently terminated, and a malformed fetus was delivered with facial dysmorphism. aCGH was applied on the DNA extracted from cord blood. Polymorphic DNA marker analysis was applied on the DNAs extracted from placenta and parental bloods. aCGH confirmed a 3.19-Mb 14q32.13-q32.2 deletion or arr 14q32.13q32.2 (96,151,751-99,341,476) × 1.0 [GRCh37 (hg19)] encompassing 10 Online Mendelian Inheritance in Man (OMIM) genes of TCL1B, TCL1A, TUNAR, BDKRB2, BDKRB1, ATG2B, GSKIP, AK7, PAPOLA and VRK1. Polymorphic DNA marker analysis confirmed a paternal origin of a de novo interstitial distal 14q deletion. Determination of the paternal origin of a prenatally detected de novo interstitial distal 14q deletion by polymorphic DNA marker analysis in this case is significant, and the information acquired is useful for genetic counseling, especially when amniocentesis is performed because of an advanced maternal age.


Analysis of Taste Sensitivities in App Knock-In Mouse Model of Alzheimer's Disease.
Some studies have reported a decline in taste sensitivities in patients with Alzheimer's disease. However, the detail remains unknown. We investigated the effect of cognitive impairment on taste sensitivity using an App knock-in mouse model of Alzheimer's disease. Behavioral assays, a brief access test, and a 48 h two-bottle preference test, to assess taste sensitivities were started from 12 months of age in mice that were confirmed to have impaired cognition. In the assays, there was no significant difference in taste sensitivities between wild type and App knock-in mice. Additionally, no apparent difference was observed in the expression of taste markers in their taste bud cells. We concluded that cognitive impairment might not greatly affect taste sensitivity.


Lack of genomic rearrangements involving the aromatase gene CYP19A1 in breast cancer.
Increased intratumoral expression of aromatase, the key enzyme for estrogen biosynthesis, is predicted to be of critical importance in the development of breast cancer. Recently, several germline rearrangements at 15q21 have been shown to cause overexpression of the aromatase gene CYP19A1 and resulting aromatase excess syndrome. To determine whether submicroscopic genomic rearrangements at 15q21 are involved in aromatase overexpression in breast cancer tissues, we investigated copy-number alterations in genomic DNA obtained from 44 tumor samples. Comparative genomic hybridization analysis identified no deletion or duplication at 15q21 in the 44 samples. These results, in conjunction with previous data, indicate that aromatase overexpression in breast cancer tissues is likely to result from a promoter switch of CYP19A1 and/or accumulation of CYP19A1-expressing cells, rather than from cryptic transactivation of CYP19A1 because of genomic rearrangements at 15q21.


Plant-driven removal of heavy metals from soil: uptake, translocation, tolerance mechanism, challenges, and future perspectives.
Increasing heavy metal (HM) concentrations in the soil have become a significant problem in the modern industrialized world due to several anthropogenic activities. Heavy metals (HMs) are non-biodegradable and have long biological half lives; thus, once entered in food chain, their concentrations keep on increasing through biomagnification. The increased concentrations of heavy metals ultimately pose threat on human life also. The one captivating solution for this problem is to use green plants for HM removal from soil and render it harmless and reusable. Although this green technology called phytoremediation has many advantages over conventional methods of HM removal from soils, there are also many challenges that need to be addressed before making this technique practically feasible and useful on a large scale. In this review, we discuss the mechanisms of HM uptake, transport, and plant tolerance mechanisms to cope with increased HM concentrations. This review article also comprehensively discusses the advantages, major challenges, and future perspectives of phytoremediation of heavy metals from the soil.


In vitro promoting activity of human interferon β production by extracts of marine algae from Japan.
Two hundreds species of marine algae were investigated for in vitro promoting activity of human interferon β (IFN-β) production by poly(I:C)-induced human osteosarcoma cell line, MG-63. A brown alga, Sargassum hemipyllum promoted most its activity, showing more than 11-fold. When we attempted to partially purify the active substances by particular two-step chromatography, two peaks of active fractions were obtained. These fractioned materials exhibited the heat-stable and non-cytotoxic characters with the molecular weight less than 3000.


Reconstituting depolarization-induced calcium release.
Researchers develop experimental platform that could be used to evaluate mutations and screen drugs for skeletal muscle diseases.


Good hydrolysis activity on raffinose family oligosaccharides by a novel α-galactosidase from Tremella aurantialba.
An α-galactosidase designated as TAG was purified from the dried fruit bodies of Tremella aurantialba with 182.5-fold purification. The purification procedure involved ion exchange chromatography on Q-sepharose, DEAE-Cellulose, and Mono Q and gel filtration by FPLC on Superdex 75. The purified α-galactosidase was a monomeric protein with a molecular mass of 88 kDa. The optimal pH of TAG was 5.0 and more than 60% of the original enzyme activity remained at pH 2.0 and 3.0. Its optimal temperature was 54 °C with good thermo-stability, 30.8% of the original activity was retained after exposure to a temperature of 70 °C for 1 h. The metal ions Hg2+, Cu2+, Fe3+ and Mg2+ strongly inhibited the enzyme activity. The enzyme activity was found to be inhibited by N-bromosuccinimide indicating that tryptophan was essential to the catalytic activity of α-galactosidase. The enzyme completely hydrolysed stachyose and partially hydrolysed raffinose to galactose at 50 °C within 6 h as detected by thin layer chromatography and the dinitrosalicylic acid method and the content of reducing sugar reached 4.36 mg/mL.


Controlled surface topography regulates collective 3D migration by epithelial-mesenchymal composite embryonic tissues.
Cells in tissues encounter a range of physical cues as they migrate. Probing single cell and collective migratory responses to physically defined three-dimensional (3D) microenvironments and the factors that modulate those responses are critical to understanding how tissue migration is regulated during development, regeneration, and cancer. One key physical factor that regulates cell migration is topography. Most studies on surface topography and cell mechanics have been carried out with single migratory cells, yet little is known about the spreading and motility response of 3D complex multi-cellular tissues to topographical cues. Here, we examine the response to complex topographical cues of microsurgically isolated tissue explants composed of epithelial and mesenchymal cell layers from naturally 3D organized embryos of the aquatic frog Xenopus laevis. We control topography using fabricated micropost arrays (MPAs) and investigate the collective 3D migration of these multi-cellular systems in these MPAs. We find that the topography regulates both collective and individual cell migration and that dense MPAs reduce but do not eliminate tissue spreading. By modulating cell size through the cell cycle inhibitor Mitomycin C or the spacing of the MPAs we uncover how 3D topographical cues disrupt collective cell migration. We find surface topography can direct both single cell motility and tissue spreading, altering tissue-scale processes that enable efficient conversion of single cell motility into collective movement.


Achieving consistent image quality with dose optimization in 64-row multidetector computed tomography prospective ECG gated coronary calcium scoring.
To evaluate the clinical value of a body mass index (BMI) based tube current (mA) selection method for obtaining consistent image quality with dose optimization in MDCT prospective ECG gated coronary calcium scoring. A formula for selecting mA to achieve desired image quality based on patient BMI was established using a control group (A) of 200 MDCT cardiac patients with a standard scan protocol. One hundred patients in Group B were scanned with this BMI-dependent mA for achieving a desired noise level of 18 HU at 2.5 mm slice thickness. The CTDIvol and image noise on the ascending aorta for the two groups were recorded. Two experienced radiologists quantitatively evaluated the image quality using scores of 1-4 with 4 being the highest. The image quality scores had no statistical difference (P = 0.71) at 3.89 ± 0.32, 3.87 ± 0.34, respectively, for groups A and B of similar BMI. The image noise in Group A had linear relationship with BMI. The image noise in Group B using BMI-dependent mA was independent of BMI with average value of 17.9 HU and smaller deviations for the noise values than in Group A (2.0 vs. 2.9 HU). There was a 35% dose reduction with BMI-dependent mA selection method on average with the lowest effective dose being only 0.35 mSv for patient with BMI of 18.3. A quantitative BMI-based mA selection method in MDCT prospective ECG gated coronary calcium scoring has been proposed to obtain a desired and consistent image quality and provide dose optimization across patient population.


The potential for efficacy of the modified (ICP 34.5(-)) herpes simplex virus HSV1716 following intratumoural injection into human malignant glioma: a proof of principle study.
We have previously demonstrated the safety of intratumoural administration of the selectively replication-competent herpes simplex virus mutant HSV1716 in patients with high-grade glioma (HGG). Here we show its potential for efficacy by demonstrating that the virus survives and replicates when injected into the tumours of patients. Since HSV replication is a cytolytic process it must result in tumour cell killing. Twelve patients with biopsy-verified HGG received an intratumoural injection of 10(5) plaque-forming units (p.f.u.) of HSV1716. Four to 9 days after inoculation, tumours were removed and assayed for evidence of viral replication. In two patients, HSV1716, in excess of the input dose was recovered from the injection site. HSV DNA was detected by PCR at the sites of inoculation in 10 patients and at distal tumour sites in four. HSV-specific antigen was detected in tumour tissue from two patients. In five patients an immunological response to HSV1716, as detected by changes in levels of IgG and IgM, was demonstrated. This study demonstrates that HSV1716 replicates in HGG without causing toxicity in both HSV-seropositive and -seronegative patients.


Proposal of De Novo Antigen Test for COVID-19: Ultrasensitive Detection of Spike Proteins of SARS-CoV-2.
Polymerase chain reaction (PCR)-based antigen tests are technically difficult, time-consuming, and expensive, and may produce false negative results requiring follow-up confirmation with computed tomography. The global coronavirus disease 2019 (COVID-19) pandemic has increased the demand for accurate, easy-to-use, rapid, and cost-effective antigen tests for clinical application. We propose a de novo antigen test for diagnosing COVID-19 using the combination of sandwich enzyme-linked immunosorbent assay and thio-nicotinamide adenine dinucleotide (thio-NAD) cycling. Our test takes advantage of the spike proteins specific to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus. The limit of detection of our test was 2.3 × 10-18 moles/assay. If the virus has ~25 spike proteins on its surface, our method should detect on the order of 10-20 moles of virus/assay, corresponding to ~104 copies of the virus RNA/assay. The detection sensitivity approaches that of PCR-based assays because the average virus RNA load used for PCR-based assays is ~105 copies per oro- or naso-pharyngeal swab specimen. To our knowledge, this is the first ultrasensitive antigen test for SARS-CoV-2 spike proteins that can be performed with an easy-to-use microplate reader. Sufficient sensitivity can be achieved within 10 min of thio-NAD cycling. Our antigen test allows for rapid, cost-effective, specific, ultrasensitive, and simultaneous multiple measurements of SARS-CoV-2, and has broad application for the diagnosis for COVID-19.


Characterization by Scatchard plots of monoclonal antibody enzyme conjugates directed against alpha-1-foetoprotein.
A radioimmunoassay was used to investigate the affinity of 2 monoclonal antibodies against human alpha-1-foetoprotein before and after conjugation with horseradish peroxidase. The equilibrium constant of the high-affinity antibody was reduced 10-fold whereas it remained unaffected in the low-affinity antibody. With the Scatchard diagram, quantification of unlabelled antibodies in the unpurified conjugate mixture is also possible if antibody affinity is changed by the coupling procedure.


The effects of water availability on the life history of the desert snail,Trochoidea seetzeni : An experimental field manipulation.
In the Negev desert of Israel, the pulmonate land snailTrochoidea seetzeni is active, grows and reproduces in the month following the torrential winter rains. Thereafter, these snails estivate until the following year's rains. By experimental supplementation of water in the field, we examined the ability of these snails to alter their life histories. Specifically, we measured changes in feeding activity, overall activity levels, movement patterns and reproductive output. We found that snails on artificially-watered plots stopped feeding on higher plants and fed on soil-surface algae growing as a result of increased water availability. Most snails moved onto the watered plots, and reduced the total amount of movement once there, in response to increased food availability. Snails on the watered plots grew more and remained active for a longer period into the summer months than did snails on control plots. However, we found little evidence for the ability of these snails to alter their reproductive outputs in response to increased water (and hence food) availability. There was also no carry-over effect of high water availability in one season enhancing reproduction in the following season. We postulate that the low level of phenotypic plasticity recorded here is a consequence of an adaptive conservative strategy to maintain a high reproductive output under extremely unpredictable desert conditions.


Disruption of mTORC1 rescues neuronal overgrowth and synapse function dysregulated by Pten loss.
Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a negative regulator of AKT/mTOR signaling pathway. Mutations in PTEN are found in patients with autism, epilepsy, or macrocephaly. In mouse models, Pten loss results in neuronal hypertrophy, hyperexcitability, seizures, and ASD-like behaviors. The underlying molecular mechanisms of these phenotypes are not well delineated. We determined which of the Pten loss-driven aberrations in neuronal form and function are orchestrated by downstream mTOR complex 1 (mTORC1). Rapamycin-mediated inhibition of mTORC1 prevented increase in soma size, migration, spine density, and dendritic overgrowth in Pten knockout dentate gyrus granule neurons. Genetic knockout of Raptor to disrupt mTORC1 complex formation blocked Pten loss-mediated neuronal hypertrophy. Electrophysiological recordings revealed that genetic disruption of mTORC1 rescued Pten loss-mediated increase in excitatory synaptic transmission. We have identified an essential role for mTORC1 in orchestrating Pten loss-driven neuronal hypertrophy and synapse formation.


Cell membrane polymorphisms coded for in the HLA-D/DR region. I. Relation between D and DR.
The relation of HLA-D and -DR determinants was studied in Dutch Caucasoids. The recognition of subgroups of DR4, DR5, and DR7, and the specificities LB12 and LB13 are described. Phenotype and gene frequencies and a Hardy--Weinberg analysis of DR and local (LB) B-cell groups are given. Excellent correlation between D and DR typing was obtained when HTCs were studied by selected B-cell antisera. When the same sera were used to type a panel of D typed cells, the correlation was decreased (with the exception of DR3 and Dw3). In the case of discrepancies the DR specificity, but not the corresponding D specificity, always could be found and not the other way around. The data fit best the assumption that HLA-D and -DR are carried by the same molecule, although they might be different determinants on this molecule. A number of possible explanations for the observed discrepancies has been given.


Secular Trends in Gastric and Esophageal Cancer Attributable to Dietary Carcinogens From 1990 to 2019 and Projections Until 2044 in China: Population-Based Study.
Little is known about trends in or projections of the disease burden of dietary gastric and esophageal cancer (GEC) in China. We aim to report GEC deaths and disability-adjusted life years (DALYs) from 1990 to 2019, predict them through 2044, and decompose changes in terms of population growth, population aging, and epidemiological changes. We retrieved dietary GEC data from the Global Burden of Disease (GBD) online database and used joinpoint regression and age-period-cohort models to analyze trends in dietary GEC deaths and DALYs from 1990 to 2019 in China. We used a Bayesian age period cohort model of integrated nested Laplace approximations to predict the disease burden of GEC through 2044 and obtained the estimated population of China from 2020 to 2050 from the Global Health Data Exchange website. Finally, we applied a recently developed decomposition method to attribute changes between 2019 and 2044 to population growth, population aging, and epidemiological changes. The summary exposure values and age-standardized rates decreased significantly from 1990 to 1999, with percentage changes of -0.06% (95% CI -0.11% to -0.02%) and -0.05% (95% CI -0.1% to -0.02%), respectively. From 1990 to 2019, for dietary esophageal cancer, the percentage change in age-standardized mortality rate (ASMR) was -0.79% (95% CI -0.93% to -0.58%) and the percentage change in age-standardized DALY rate (ASDR) was -0.81% (95% CI -0.94% to -0.61%); these were significant decreases. For dietary stomach cancer, significant decreases were also observed for the percentage change in ASMR (-0.43%, 95% CI -0.55% to -0.31%) and the percentage change in ASDR (-0.47%, 95% CI -0.58% to -0.35%). In addition, data from both the joinpoint regression and annual percentage change analyses demonstrated significantly decreasing trends for the annual percentage change in ASMR and ASDR for GEC attributable to dietary carcinogens. The overall annual percentage change (net drift) was -5.95% (95% CI -6.25% to -5.65%) for dietary esophageal cancer mortality and -1.97% (95% CI -2.11% to -1.83%) for dietary stomach cancer mortality. Lastly, in 2044, dietary esophageal cancer deaths and DALYs were predicted to increase by 192.62% and 170.28%, respectively, due to age structure (121.58% and 83.29%), mortality change (76.81% and 92.43%), and population size (-5.77% and -5.44%). In addition, dietary stomach cancer deaths and DALYs were predicted to increase by 118.1% and 54.08%, with age structure, mortality rate change, and population size accounting for 96.71% and 53.99%, 26.17% and 3.97%, and -4.78% and -3.88% of the change, respectively. Although the predicted age-standardized rates of mortality and DALYs due to dietary GEC show downward trends, the absolute numbers are still predicted to increase in the next 25 years due to rapid population aging in China.


Secondary in-hospital epidemiological investigation after an outbreak of Pseudomonas aeruginosa ST357.
The secondary in-hospital epidemiological investigation for drug-resistant Pseudomonas aeruginosa infections was conducted to evaluate the in-hospital situation and identify any associations between exoenzyme genotypes and other genotypes and antimicrobial resistance characteristics, at the University Hospital in Kyoto, Japan, following a reported outbreak of antimicrobial-resistant P. aeruginosa ST357 between 2005 and 2014. Twelve of the 546 P. aeruginosa isolates collected during the follow-up period were resistant to more than two classes of antimicrobials. All isolates were resistant to fluoroquinolones and 8 (66.7%) showed carbapenem resistance. None of the isolates fulfilled the clinical criteria for multidrug-resistant P. aeruginosa. All isolates were metallo-β-lactamase test-negative. Among five exoS (-)exoU (+) isolates, three possessing a class 1 integron with gene cassette aadB + cmlA6 were classified as ST357, and one isolate containing a class 1 integron with aacA31 was ST235. Collectively, the second survey results confirm that the initial outbreak is currently undergoing convergence. By combining data from the first and second surveys, we showed that prevalent STs such as ST357 and ST235 are associated with fluoroquinolone resistance, class 1 integron-associated resistance to β-lactams and aminoglycosides, and cytotoxic exoU (+) genotypes. With the current worldwide spread of ST357 and ST235 isolates, it is important to evaluate epidemiological trends for high-risk P. aeruginosa isolates by continuous hospital monitoring.


Benthic foraminifera as bioindicators for the heavy metals in the severely polluted Hurghada Bay, Red Sea coast, Egypt.
Twenty-nine sediment samples were collected from the Hurghada Bay, a heavily polluted bay on the Red Sea of Egypt, to inspect the environmental quality status and anthropogenic consequences on benthic foraminifera. Some foraminiferal species showed deformations in their apertures and coiling directions as a response to environmental stresses. In addition, the FoRAM index, an index used for evaluating the growth of coral reefs, indicated a hazard in the proximity of nearshore stations. To elucidate the relationships between the biological response and chemistry of sediments, eight heavy metals concentrations (Cu, Cd, Zn, Pb, As, Cr, Ni, and Mn) were analyzed using inductively coupled plasma-atomic emission spectrometers (ICP-AES). Interestingly, two groups of benthic foraminiferal associations were illustrated using multivariate statistical analyses. Group I have extremely high heavy metal concentrations, an enriched total organic matter (TOM)%, high deformation percentages, and mud content. Moreover, it is dominated by Ammonia tepida which is regarded as an opportunistic species. Group II includes low to moderately polluted stations, highly enriched living foraminiferal assemblages, and is dominated by the sensitive rotaliids Neorotalia calcar and Amphistegina lobifera. Alternatively, four geochemical indices, EF, CF, Igeo, and PLI, are used to assess the contamination level that shown ominous spots for the nearshore stations of the Hurghada Bay. The pollution indices (HQ and HI) were also conducted to evaluate the risks of carcinogenic heavy metals on human health. Our findings demonstrated that ingestion and dermal exposure have greater carcinogenic hazards for adults and children than inhalation. The lifetime carcinogenic risk (LCR) is significantly higher than the permissible limit and follows this order: Pb > As > Cr > Cd > Ni. To that end, developing strategies to lessen the negative impact of pollution on human health and/or the Red Sea's biodiversity is an inevitable issue in the present day and future.


Validation Study of the PALCOM Scale of Complexity of Palliative Care Needs: A Cohort Study in Advanced Cancer Patients.
In a patient-centred model of care, referral to early palliative care (EPC) depends on both the prognosis and the complexity of care needs. The PALCOM scale is a 5-domain multidimensional assessment tool developed to identify the level of complexity of palliative care needs of cancer patients. The aim of this study was to validate the PALCOM scale. We conducted a prospective cohort study of cancer patients to compare the PALCOM scale and expert empirical assessment (EA) of the complexity of palliative care needs. The EA had to categorise patients according to their complexity, considering that medium to high levels required priority attention from specialist EPC teams, while those with low levels could be managed by non-specialist teams. Systematically collected multidimensional variables were recorded in an electronic report form and stratified by level of complexity and rating system (PALCOM scale versus EA). The correlation rank (Kendall's tau test) and accuracy test (F1-score) between the two rating systems were analysed. ROC curve analysis was used to determine the predictive power of the PALCOM scale. A total of 283 advanced cancer patients were included. There were no significant differences in the frequency of the levels of complexity between the EA and the PALCOM scale (low 22.3-23.7%; medium 57.2-59.0%; high 20.5-17.3%). The prevalence of high symptom burden, severe pain, functional impairment, socio-familial risk, existential/spiritual problems, 6-month mortality and in-hospital death was significantly higher (p < 0.001) at the high complexity levels in both scoring systems. Comparative analysis showed a high correlation rank and accuracy between the two scoring systems (Kendall's tau test 0.81, F1 score 0.84). The predictive ability of the PALCOM scale was confirmed by an area under the curve in the ROC analysis of 0.907 for high and 0.902 for low complexity. In a patient-centred care model, the identification of complexity is a key point to appropriate referral and management of shared care with EPC teams. The PALCOM scale is a high precision tool for determining the level of complexity of palliative care needs.


Changes in physical and mental health in Swedish schoolchildren in the digital age.
Poor physical and mental health are common and increasing in Swedish children. The aim was to investigate significant changes in health and digital habits in fourth grade and ninth grade pupils in 2020 compared to 2011. A validated questionnaire was conducted in 2011 and 2020 among schoolchildren in fourth (10 years) and ninth grade (15 years) in Sweden. The study population consisted of 196 students in 2020 and 254 students in 2011. A significant increase was seen in self-reported sleeping difficulties, tiredness, life dissatisfaction and dissatisfaction with academic abilities among fourth grade children and a sixfold increase in their refusal to go to school. Students in fourth grade also reported that they spent more time on social media and had a reduced interest in sports. Both students in fourth and ninth grade described an increase in attacks of anger and unhappiness, as well as decreased time with family. We conclude that fourth grade children have more physical and mental health symptoms in 2020 compared to 2011. The ninth grade students showed a high frequency of physical and mental symptoms in 2011, which remained high in 2020. One possible explanation may be that digital habits have increased.


Posttraumatic primary brainstem haematoma.
Nine cases of posttraumatic primary brain stem haematoma are described. All cases presented ocular and vegetative symptoms. Hyperextension was regarded as the most likely mechanism of injury. All patients were treated conservatively; half of them with a good outcome.


Functional expression of the gene encoding cytidine triphosphate synthetase from Plasmodium falciparum which contains two novel sequences that are potential antimalarial targets.
CTP synthetase (E C 188.8.131.52 UTP: ammonia ligase (ADP-forming)) catalyses the formation of CTP from UTP and, in the human parasite Plasmodium falciparum, is the sole source of cytidine nucleotides. It is thus a potential chemotherapeutic target, especially as the gene sequence indicated that the encoded GAT-domain of the enzyme contains two extended peptide segments (42aa and 223aa as compared to the host enzyme). Here, we circumvent the codon usage problems associated with the high A/T content of the P. falciparum sequence, especially evident in sequences encoding the extra peptides, to successfully express active recombinant P. falciparum CTP synthetase using preferred E. coli codons. This partially synthetic gene produced recombinant enzyme, containing the additional segments, which was functionally assayed for activity in vitro. We also show the native enzyme contains the additional peptides using immunoblots with antibodies derived from the recombinant protein. Confocal microscopy, using antibodies to the recombinant protein, provided evidence that the enzyme is expressed in vivo. This establishes for the first time that P. falciparum contain active CTP synthetase and that this enzyme contains two novel insert sequences in the functional enzyme.


Fine structure of myomyous junctions in the mouse skeletal muscles.
The reaction product of acetylcholinesterase (AChE) activity is known to be specifically localized at a neuromuscular junction and a muscle-tendon junction of the striated skeletal muscles. In addition to the two junctions, we recently found some linear precipitates due to AChE activity running transversely across a fibre of the semitendinosus, rectus abdominis, gastrocnemius, tibialis anterior and diaphragm muscles in mice. Under an electron microscope, the linear precipitates were seen at the extracellular side of the muscle fibre endings. Most of the endings contacted each other to form a junction, which has been called the 'myomyous junction (M-Mj)'. The patterns of the M-Mj were grouped into three types: (1) a junction in which all contacts were firm, without any connective tissue, and invaginated deeply; (2) the ones in which numerous collagen fibres were visible in the space between the two separate opposing muscle fibres; (3) an intermediate type between (1) and (2), i.e. a junction with partial contacts. The muscle fibre ending forming M-Mj was constructed of finger-like processes like that of a muscle-tendon junction. However, the processes of a M-Mj adhered so closely to each other that no collagen fibrils could penetrate into their folds.


Temporal effects of estradiol (E) on luteinizing hormone-releasing hormone (LHRH) and LH release in castrated male sheep.
We tested the hypothesis that rapidly expressed inhibitory effects of estradiol (E) on luteinizing hormone (LH) release in the male are attributable, in part, to suppression of luteinizing hormone-releasing hormone (LHRH) release. Hypophyseal-portal cannulated, castrated male sheep were infused with E (15 ng/kg/hr) or vehicle. Portal and jugular blood samples were collected at 10-min intervals for 4 hr before, and for either 12 hr (E, n = 4; vehicle, n = 4) or 24 hr (E, n = 8; vehicle, n = 3) after the start of infusion. In animals sampled for 16 hr, temporal changes in both LHRH and LH were assessed. In animals sampled for 28 hr, only LH data were analyzed. Before either the 12-hr or 24-hr infusion, LHRH and/or LH mean concentrations, pulse amplitude and interpulse interval (IPI) did not differ between E- and vehicle-infused animals. In animals sampled for 16 hr, no effects of time or steroid x time interactions were detected for mean LHRH and LHRH pulse amplitude; however, both were greater (P < 0.01) in vehicle-infused than in E-infused males. LHRH IPI was unaffected by infusion. In contrast, both mean LH and LH pulse amplitude declined (P < 0.01) within 4-8 hr after the start of E infusion, whereas mean LH IPI was unaffected. In animals sampled for 28 hr, an effect of time (P < 0.01) and a steroid x time interaction (P < 0.01) was detected for mean LH, and there was an effect of time (P < 0.01) on LH pulse amplitude. Mean LH IPI was not affected. Our results show that in male sheep E rapidly reduces LH release in the absence of a detectable change in LHRH release.


Formation of diastereoisomeric 3a-hydroxypyrroloindoles from a tryptophan residue analog mediated by iron (II)-EDTA and L-ascorbate.
Oxygenation of a tryptophan residue analog by ascorbate in the presence of catalytic amounts of iron(II) and ethylenediaminetetraacetic acid (EDTA) has been studied. Under physiological conditions, reaction of the tryptophan derivative (N-t-butoxycarbonyl-L-tryptophan) with Fe(II)-EDTA and ascorbate resulted mainly in the oxygenation of the indole moiety of the substrate. In this reaction, cis and trans diastereoisomeric alcohols 3a-hydroxy-1-t-butoxycarbonyl-1,2,3,3a,8,8a-hexahydropyrrolo[2,3- b]indoles have been successfully identified in the metal-catalyzed free radical oxidation of indole compounds. Hydroxylation at C-5 and C-6 and a ring opening reaction between C-2 and C-3 have also been confirmed. The reaction of Fe(II)-EDTA/ascorbate with the tryptophan derivative was apparently nonselective with regard to position and was significantly suppressed by the hydroxyl radical scavengers (mannitol and dimethylsulfoxide), suggesting the participation of the hydroxyl radical as the actual oxidizing species.


The hypotensive effect of the nitric monoxide donor Oxacom at different routs of its administration to experimental animals.
Earlier it has been found that the hypotensive drug Oxacom containing binuclear dinitrosyl iron complexes (B-DNIC) with glutathione can effectively decrease, as a nitric monooxide (NO) donor, the mean arterial pressure (МАР) in rats upon intravenous bolus injection in the form of an aqueous solution (Chazov et al., 2012). The aim of this study was to investigate the hypotensive effects of Oxacom administered to experimental rats by intravenous, intramuscular, subcutaneous, intraperitoneal, intragastric, rectal routes.MAP and heart rate (HR) were measured with the help of arterial catheters equipped with tensometric sensors. Oxacom was administered to rats at the dose of 2.0 μmole of B-DNIC/kg. The concentration of paramagnetic mononuclear protein-bound DNIC (М-DNIC) formed in the blood and tissues of various internal organs of the rat was determined by the EPR method. Upon subcutaneous, intramuscular or intraperitoneal administration of Oxacom, the maximum amplitude of the МАР decrease varies from 30% to 70%, respectively, in comparison with the corresponding parameter for the intravenously injected Oxacom. Another difference is the lack of the fast phase in the initial stage of the МАР decrease and the longer persistence of protein-bound M-DNIC formed in the circulating blood after intramuscular, subcutaneous or intraperitoneal administration of Oxacom. Thus, the NO donor Oxacom exerts pronounced hypotensive effects on rats not only upon intravenous, but also upon intramuscular, subcutaneous or intraperitoneal administration.


Provider confidence in the telemedicine spine evaluation: results from a global study.
To utilize data from a global spine surgeon survey to elucidate (1) overall confidence in the telemedicine evaluation and (2) determinants of provider confidence. Members of AO Spine International were sent a survey encompassing participant's experience with, perception of, and comparison of telemedicine to in-person visits. The survey was designed through a Delphi approach, with four rounds of question review by the multi-disciplinary authors. Data were stratified by provider age, experience, telemedicine platform, trust in telemedicine, and specialty. Four hundred and eighty-five surgeons participated in the survey. The global effort included respondents from Africa (19.9%), Asia Pacific (19.7%), Europe (24.3%), North America (9.4%), and South America (26.6%). Providers felt that physical exam-based tasks (e.g., provocative testing, assessing neurologic deficits/myelopathy, etc.) were inferior to in-person exams, while communication-based aspects (e.g., history taking, imaging review, etc.) were equivalent. Participants who performed greater than 50 visits were more likely to believe telemedicine was at least equivalent to in-person visits in the ability to make an accurate diagnosis (OR 2.37, 95% C.I. 1.03-5.43). Compared to in-person encounters, video (versus phone only) visits were associated with increased confidence in the ability of telemedicine to formulate and communicate a treatment plan (OR 3.88, 95% C.I. 1.71-8.84). Spine surgeons are confident in the ability of telemedicine to communicate with patients, but are concerned about its capacity to accurately make physical exam-based diagnoses. Future research should concentrate on standardizing the remote examination and the development of appropriate use criteria in order to increase provider confidence in telemedicine technology.


[Study of functional interhemispheric asymmetry in children in the context of electrophysiological markers and the efficiency of neurological rehabilitation].
The paper presents neurological findings of children and adolescents, which included the determination of the individual profile of cerebral asymmetry by 3 spheres: motor, sensory, and mental, as well as electrophysiological measurement. To develop biology teaching methods by taking into account the individual profile of pupils' brain asymmetry is an aim of this study.


The small scale functional topology of movement control: Hierarchical organization of local activity anticipates movement generation in the premotor cortex of primates.
How neurons coordinate their collective activity for behavioural control is an open question in neuroscience. Several studies have progressively proven, on various scales, that the patterns of neural synchronization change accordingly with behavioural events. However, the topological features of the neural dynamics that underlie task-based cognitive decisions on the small scale level are not understood. We analysed the multiunit activity (MUA) from a multielectrode (96 channels) array of the dorsal premotor cortex (PMd) in rhesus monkeys during a countermanding reaching task. Within the framework of graph theory, we found that in the local PMd network motor execution is preceded by the emergence of hubs of anti-correlation that are organized in a hierarchical manner. Conversely, this organization is absent when monkeys correctly inhibit programmed movements. Thus, we interpret the presence of hubs as reflecting the readiness of the motor plan and the irrevocable signature of the onset of the incoming movement.


Role of protease activated receptor-2 in lymph node metastasis of uterine cervical cancers.
Protease activated receptor-2 (PAR-2) has been implicated in cellular proliferation, invasion and metastasis in various tumors. Lymph node metastasis is an important patient prognostic factor for uterine cervical cancers. This prompted us to study the role of PAR-2 in lymph node metastasis of uterine cervical cancers. Thirty patients underwent surgery for uterine cervical cancers. PAR-2 histoscores and mRNA levels were determined by immunohistochemistry and real-time reverse transcription-polymerase chain reaction, respectively. Patient prognosis was analyzed with a 48-month survival rate. PAR-2 histoscores and mRNA levels significantly (P < 0.05) increased in 12 of 30 metastatic lymph node lesions from the corresponding primary tumor. The 48-month survival rate of the 12 patients with increased PAR-2 levels in metastatic lymph nodes was 42%, while the rate of the other 18 patients with no change in PAR-2 levels was 82%, regardless of histopathological type. PAR-2 might work on lymph node metastasis of uterine cervical cancers, and is considered to be a novel prognostic indicator for uterine cervical cancers.


Neurotensin interacts with carbachol, secretin, and caerulein in the stimulation of the exocrine pancreas of the rat in vitro.
In the present investigation the effect of neurotensin on pancreatic secretion of isolated pancreatic lobules from the rat was examined. We found a dose- and time-dependent stimulation of amylase release beginning with a concentration of 10(-9) M neurotensin. This response was potentiated by the cholinergic agonist carbachol, the gastrointestinal peptide secretin, and the CCK analogue caerulein. As we found neurotensin-immunoreactive nerves within the pancreas and as neurotensin-like immunoreactivity is present in the circulation (found previously), neurotensin may well be a further peptide taking part in the regulation of exocrine pancreatic secretion either as a hormone or a neurotransmitter. Neurotensin would then cooperate with cholinergic mechanisms, secretin, and CCK.


Verification system for intensity-modulated radiation therapy with scintillator.
In the preparation of intensity-modulated radiation therapy (IMRT), patient-specific verification is widely employed to optimize the treatment. To accurately estimate the accumulated dose and obtain the field-by-field or segment-by-segment verification, an original IMRT verification tool using scintillator light and an analysis workflow was developed in this study. The raw light distribution was calibrated with respect to the irradiated field size dependency and light diffusion in the water. The calibrated distribution was converted to dose quantity and subsequently compared with the results of the clinically employed plan. A criterion of 2-mm dose-to-agreement and 3% dose difference was specified in the gamma analysis with a 10% dose threshold. By applying the light diffusion calibration, the maximum dose difference was corrected from 7.7 cGy to 3.9 cGy around the field edge for a 60 cGy dose, 7 × 7 cm2 irradiation field, and 10 MV beam energy. Equivalent performance was confirmed in the chromodynamic film. The average dose difference and gamma pass rate of the accumulated dose distributions in six patients were 0.8 ± 4.5 cGy and 97.4%, respectively. In the field-by-field analysis, the average dose difference and gamma pass rate in seven fields of Patient 1 were 0.2 ± 1.2 cGy and 93.9%, respectively. In the segment-by-segment analysis, the average dose difference and gamma pass rate in nine segments of Patient 1 and a 305° gantry angle were - 0.03 ± 0.2 cGy and 93.9%, respectively. This system allowed the simultaneous and independent analysis of each field or segment in the accumulated dose analysis.


Method for Isolation of Myxozoan Proliferative Stages from Fish at High Yield and Purity: An Essential Prerequisite for In Vitro, In Vivo and Genomics-Based Research Developments.
Myxozoans are a diverse group of microscopic cnidarian parasites and some representatives are associated with important diseases in fish, in both marine and freshwater aquaculture systems. Research on myxozoans has been largely hampered by the inability to isolate myxozoan parasites from their host tissues. In this study, we developed and optimized a method to isolate the myxozoan proliferative stages of different size and cellularity from fish blood, using DEAE-cellulose ion exchange chromatography. We optimized several parameters and obtained 99-100% parasite purity, as well as high survival and infectivity. Using polyclonal pan-carp blood cell-specific antibodies, we further developed a rapid cytometric assay for quantification of the proliferative stages, not only in highly concentrated DEAE-C isolates but also in dilute conditions in full blood. Early developmental stages of myxozoans are key to parasite proliferation, establishment, and pathology in their hosts. The isolation of these stages not only opens new possibilities for in vivo and in vitro studies, but also for obtaining purified DNA and protein extracts for downstream analyses. Hence, we provide a long-desired tool that will advance the functional research into the mechanisms of host exploitation and immune stimulation/evasion in this group, which could contribute greatly to the development of therapeutic strategies against myxozoans.


Switching from reservoir to matrix systems for the transdermal delivery of fentanyl: a prospective, multicenter pilot study in outpatients with chronic pain.
Matrix systems for transdermal fentanyl will replace the reservoir systems in Europe. In an industry-independent, prospective, multicenter pilot study, 46 outpatients with chronic pain were asked to assess pain intensity, sleep interference, adverse events, and multiple secondary parameters during administration of the last reservoir and the first two matrix patches. There was no difference in pain intensity, sleep interference, and the rate of adverse events between both systems. Self assessment on a 6-step numeric rating scale (1 = very good, 6 = insufficient) comparing the two systems (reservoir vs. matrix) showed that skin compatibility (2.6 vs. 1.5), adhesive properties (3.2 vs. 1.8), wearability/comfort (2.8 vs. 1.5), and general satisfaction (2.5 vs. 1.8) improved significantly with the new matrix technology. At study endpoint, 91% of patients preferred the matrix system for future use. The new fentanyl matrix system is characterized by a high level of general satisfaction, ease of use, patient acceptance, and improved skin compatibility. Reservoir and matrix systems appear to have comparable efficacy and safety so that outpatients can be switched directly from the reservoir to the matrix system without difficulties and new dose titration.


Major niche transitions in Pooideae correlate with variation in photoperiodic flowering and evolution of CCT domain genes.
The external cues that trigger timely flowering vary greatly across tropical and temperate plant taxa, the latter relying on predictable seasonal fluctuations in temperature and photoperiod. In the grass family (Poaceae) for example, species of the subfamily Pooideae have become specialists of the northern temperate hemisphere, generating the hypothesis that their progenitor evolved a flowering response to long days from a short-day or day-neutral ancestor. Sampling across the Pooideae, we found support for this hypothesis, and identified several secondary shifts to day-neutral flowering and one to short-day flowering in a tropical highland clade. To explain the proximate mechanisms for the secondary transition back to short-day-regulated flowering, we investigated the expression of CCT domain genes, some of which are known to repress flowering in cereal grasses under specific photoperiods. We found a shift in CONSTANS 1 and CONSTANS 9 expression that coincides with the derived short-day photoperiodism of our exemplar species Nassella pubiflora. This sets up the testable hypothesis that trans- or cis-regulatory elements of these CCT domain genes were the targets of selection for major niche shifts in Pooideae grasses.


Cumulative effect of zinc oxide and titanium oxide nanoparticles on growth and chlorophyll a content of Picochlorum sp.
The use of nanoparticles (NPs) is of increasing significance due to their large potential for various applications. Great attention should be paid on the possible impacts of nanoparticles on the environment as large amounts of them may reach the environment by accident or voluntarily. Marine algae are potential organisms for usage in nanopollution bioremediation in aquatic system, because of their ability to adapt to long exposure to NPs. Thus, it is of prime importance to study the possible interactions of different NPs with microalgae in assessing their potential environmental risks. Most studies on potential environmental effects of ZnO and TiO2 NPs have been performed independently and following the widely accepted, standardized test systems, which had been developed for the characterization of chemicals. In this study, we have examined the cumulative effect of ZnO and TiO2 NPs on Picochlorum sp. in addition to the individual effects of these NPs over 32 days. Our results indicate that the toxicity and availability of NPs to marine algae are reduced by their aggregation and sedimentation. NPs are found to have a negative effect on algal growth and chlorophyll a concentration during the early growth stages. In contrast, the case is reversed during the late growth stages. There is no significant difference between the effect of the NPs when they are used separately and when both ZnO and TiO2 are used together in the test (P > 0.05).


Analysis of film response to electron beams using a laser scanner system developed in the laboratory.
An optical scanner system, which incorporates a He-Ne laser, photodiode detectors, and a platform for placing film, was built in the laboratory. The laser system operates at the green wavelength of 543.5 nm and functions as a scanning densitometer for measurement of optical changes in a film resulting from irradiation . The central axis electron depth dose of selected electron energies 10,12 and 14 MeV were analysed using Kodak X-Omat and Kodak Extended Dose Range (EDR2) films. The Kodak X-Omat film is routinely used for high-energy electron dose distributions in radiation therapy. The electron depth-dose measured with X-Omat film was found to agree well with standard depth-dose curves in water, obtained using an ion chamber. Conversely, the recently introduced Kodak EDR2 showed an energy dependence for electron beams, the percentage depth-dose curve shifting towards the surface for 12 and 14 MeV electron beams compared to that in water.


A western blot assay to measure cyclin dependent kinase activity in cells or in vitro without the use of radioisotopes.
We developed a quantitative method to measure the activity of cyclin-dependent kinases (Cdks) by western blotting, without radioisotopes. We prepared a recombinant protein substrate based upon the natural Cdk1 substrate, PP1Cα. By combining this substrate in a western blot method using fluorochrome based antibodies and phospho-imager analysis, we measured the Km of ATP binding to Cdk1 to be 3.5 μM. We then measured Cdk1 activity in cell extracts from interphase or mitotic cells, and demonstrated that previously identified Cdk inhibitors could be detected by this assay. Our data show that we have a safe, reliable assay to identify Cdk1 inhibitors and measure Cdk1 activity.


Effects of riluzole on P2X7R expression in the spinal cord in rat model of neuropathic pain.
Neuropathic pain is becoming an intractable health threat, with its profound effect on quality of life, thus posing a major challenge to clinical therapy. Despite the reported efficacy of riluzole in some pain models, the underlying mechanism remains largely unknown. The present study aimed to assess the effects of riluzole in a rat model of neuropathic pain induced by chronic constriction injury (CCI). Subsequent to model establishment, paw withdrawal latencies (PWLs) and the paw withdrawal mecha threshold (PWT) rapidly decreased, coupled with inhibited microglial activation and upregulated P2X7R expression in the spinal cord dorsal horn (SCDH). Following intraperitoneal administration of riluzole (4mg/kg) once daily for 5 consecutive days as from day 3 after surgery, the mechanical allodynia and thermal hyperalgesia in the hind limbs were significantly attenuated. In addition, riluzole downregulated P2X7R expression and inhibited microglial activation in SCDH. Our results indicated that riluzole is effective in alleviating neuropathic pain and inhibiting microglial activation, presumably via the downregulated P2X7R expression in SCDH.


Microglial activation and TNFalpha production mediate altered CNS excitability following peripheral inflammation.
Peripheral inflammation leads to a number of centrally mediated physiological and behavioral changes. The underlying mechanisms and the signaling pathways involved in these phenomena are not yet well understood. We hypothesized that peripheral inflammation leads to increased neuronal excitability arising from a CNS immune response. We induced inflammation in the gut by intracolonic administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) to adult male rats. To examine the excitability of the brain in vivo, we administered pentylenetetrazole (PTZ; a GABAergic antagonist) intravenously to evoke clonic seizures. Rats treated with TNBS showed increased susceptibility to PTZ seizures that was strongly correlated with the severity and progression of intestinal inflammation. In vitro hippocampal slices from inflamed, TNBS-treated rats showed increased spontaneous interictal burst firing following application of 4-aminopyridine, indicating increased intrinsic excitability. The TNBS-treated rats exhibited a marked, reversible inflammatory response within the hippocampus, characterized by microglial activation and increases in tumor necrosis factor alpha (TNFalpha) levels. Central antagonism of TNFalpha using a monoclonal antibody or inhibition of microglial activation by i.c.v. injection of minocycline prevented the increase in seizure susceptibility. Moreover, i.c.v. infusion of TNFalpha in untreated rats for 4 days also increased seizure susceptibility and thus mimicked the changes in seizure threshold observed with intestinal inflammation. Our finding of a microglia-dependent TNFalpha-mediated increase in CNS excitability provides insight into potential mechanisms underlying the disparate neurological and behavioral changes associated with chronic inflammation.


Biosensor plates detect mitochondrial physiological regulators and mutations in vivo.
The measurement of mitochondrial activity in living cells is usually not straightforward, even though it is quite important in physiological and pathophysiological processes. We describe a high-throughput method for measurement of mitochondrial oxygen consumption in living cells, based on the Becton-Dickinson Biosensor plates.


Impact of abdominal or pelvic radiotherapy on disease activity in inflammatory bowel disease: a multicentre cohort study from the GETAID.
Abdominal or pelvic radiotherapy in inflammatory bowel disease (IBD) patients raises concerns regarding the risk of worsening of underlying disease. To assess the impact of radiotherapy on IBD course. A retrospective multicentre study including IBD patients exposed to abdominal or pelvic irradiation was conducted, retrieving IBD activity by semester (6-month periods) before (from S-4 to S-1) and after (from S + 1 to S + 6) radiotherapy and IBD flare during follow-up. Sixty-one patients (32 women, mean age 59 years), with 467 patient semesters of follow-up, treated for digestive (n = 31), urinary tract (n = 23) and gynaecological cancers (n = 7) were included. Rates of IBD activity per semester were, respectively, 21% (95% CI: 16-27) from S-4 to S-1; 12% (7-19) from S + 1 to S + 3 (P = 0.15 vs S-4 to S-1) and 16% (10-25) from S + 4 to S + 6 (P = 0.45 vs S-4 to S-1). With a median follow-up of 156 weeks (interquartile range: 82-365), rates of survival without IBD flare at 1 and 3 years after radiotherapy were 82.5% (73.2-93.0) and 70.6% (58.8-84.7). Moderate-to-severe acute radiotherapy-induced gut toxicity and the absence of concomitant chemotherapy were independently associated with an increased risk of flare. Most patients with non-active IBD can be safely treated with abdominal or pelvic radiotherapy. Patients having acute gut toxicity and those without concomitant chemotherapy should be more closely monitored in the post-radiotherapy period.


Bone Using Stem Cells for Maxillofacial Bone Disorders: A Systematic Review and Meta-analysis.
Due to economic, cultural, environmental, and social factors, the prevalence of maxillofacial bone disorders varies in different parts of the world. The present meta-analysis was conducted to assess the efficacy and safety of different type of stem cells-based scaffolds and their construction methods in maxillofacial bone disorders. We searched major indexing databases, including PubMed/Medline, ISI Web of Science, Scopus, Embase, and Cochrane Central without any language, study region, or type restrictions. A systematic search of articles published up to July 2021 was done. Of the 428 studies found through initial searches, 36 met the inclusion criteria. After applying the exclusion criteria, the main properties of 32 articles on 643 animals and 4 experimental studies on 52 patients (age range from 43 to 74 years) included in this meta-analysis. Our pooled analysis showed that stem cells-based scaffolds significantly improved the bone regeneration and formation in maxillofacial bone disorders (Prevalence: 0.54; 95% CI: 0.43, 0.64, P < 00001, I2 = 90 2). According to the results of these studies, in most studies, bone marrow-derived mesenchymal stem cells (BMSCs) have been used to regenerate bone, and these cells are still the gold standard in bone tissue engineering, a growth factor that is one of the three sides of the tissue engineering triangle. Bone morphogenetic proteins (BMP) especially BMP2 and platelet-rich plasma (PRP) are the most widely used growth factor and scaffold respectively. Platelet-rich plasma (PRP) is used as a scaffold and since it contains proteins, it also used as a growth factor and can be a stimulant of ossification. It seems that the future perspective of bone tissue engineering is to use the prototyping rapid method to build a composite and patient-specific scaffold from CT and MRI images, along with genetically modified stem cells.


Synthesis, intramolecular hydrogen bonding, and biochemical studies of clitocine, a naturally occurring exocyclic amino nucleoside.
The total synthesis of clitocine [6-amino-5-nitro-4-(beta-D-ribofuranosylamino)pyrimidine] (1), a nucleoside recently isolated from the mushroom Clitocybe inversa, has been accomplished. Glycosylation of 4,6-diamino-5-nitropyrimidine (4) with 1-O-acetyl-2,3,5-tri-O-benzoyl-D-ribofuranose afforded the protected nucleoside 6-amino-5-nitro-4-[(2,3,5-tri-O-benzoyl-beta-D-ribofuranosyl) amino]pyrimidine (5) in good yield exclusively as the beta-anomer. Deprotection of 5 with NaOMe/MeOH gave 1 as an 11.5:1 mixture of the beta- and alpha-anomers, respectively. Recrystallization from MeOH, followed by chromatography, afforded 1 containing less than 1% of its alpha-anomer. X-ray crystal data revealed a planar aglycon moiety in clitocine with each oxygen atom of the nitro group intramolecularly hydrogen bonded to the hydrogen atoms of the two adjacent amino functions. Clitocine inhibited L1210 cells in vitro with an ID50 of 3 X 10(-8) M. Clitocine was also found to be a substrate and inhibitor of adenosine kinase with a Ki value of 3 X 10(-6) M.


[General screening on ICUs - the key for an effective MRSA prevention?].
The recommendations of the German Robert Koch Institute, concerning microbiological screening of newly admitted hospital patients for MRSA colonization, referred to specific risk groups such as patients admitted from long-term care facilities. New literature data indicate that a general MRSA screening policy of all incoming patients may be a cost-effective measure in intensive care units of large tertiary care hospitals.


Myocardial energy metabolism in ischemic preconditioning and cardioplegia: a metabolic control analysis.
For both, cardioplegia (CP) and ischemic preconditioning (IP), increased ischemic tolerance with reduction in infarct size is well documented. These cardioprotective effects are related to a limitation of high energy phosphate (HEP) depletion. As CP and IP have to be assumed to act by different mechanisms, their effects on myocardial HEP metabolism cannot be assumed to be identical. Therefore, a systematic analysis of myocardial HEP metabolism for both procedures and their combination was performed, addressing the question whether there are different effects on myocardial HEP metabolism by IP and CP. In this study, metabolic control analysis was used to analyze the regulation of HEP metabolism. In open chest pigs subjected to 45 min LAD occlusion (index ischemia), CP and IP preserved myocardial ATP (control (C) 0.14 +/- 0.05 micromol/g wwt; CP: 0.95 +/- 0.14, IP: 0.61 +/- 0.12; p<0.05 C vs. CP and IP) and reduced myocardial necrosis (infarct size IA/RA: C: 90.0 +/- 3.0%; CP: 0.0 +/- 0.0% but patchy necroses; IP: 5.05 +/- 2.1%; p<0.05 C vs. CP and IP). The effects on HEP metabolism, however, were different: CP acted predominantly by slowing down the breakdown of phosphocreatine (PCr) during early phases of ischemia (C: DeltaPCr 0-2 min: 5.24 +/- 0.32 micromol/g wwt; CP: DeltaPCr 0-2 min: 3.38 +/- 0.23 micromol/g wwt, p<0.05 vs. C), leaving ATP breakdown during later stages unaffected (C: DeltaATP 5-45 min: 1.77 +/- 0.11 micromol/g wwt CP: DeltaATP 5-45 min: 1.59 +/- 0.28 micromol/g wwt, n.s. vs. C). In contrast to CP, in IP PCr breakdown was even increased (IP: DeltaPCr 0-2 min: 7.06 +/- 0.34 micromol/g wwt, p<0.05 vs. C), but ATP depletion greatly attenuated (IP: DeltaATP 5-45 min: 0.48 +/- 0.10 micromol/g wwt, p<0.05 vs. C and CP). Combining IP and CP yielded an additive effect with slowing down the breakdown of both PCr (IP+CP: DeltaPCr 0-2 min: 5.09+/- 0.35 micromol/g wwt, p<0.05 vs. C and IP) and ATP (IP+CP: DeltaATP 5-45 min: 0.56 +/- 0.48 micromol/g wwt, p<0.05 vs. C and CP), resulting in a higher ATP content at the end of index ischemia (1.86 +/- 0.46 micromol/g wwt, p<0.05 vs. C, CP and IP). Compared to IP, combining IP+CP achieved also a further reduction in infarct size (IA/RA: 0.0 +/- 0.0%, p<0.05 vs IP) and--compared to CP--a disappearance of the patchy necroses. The concept of major differences in myocardial HEP metabolism during CP and IP is further supported at a molecular level by metabolic control analysis. CP but not IP slowed down the CK reaction velocity at high PCr levels. In contrast to CP exerting a continuous decline in vATPase for any given ATP level, in IP myocardium ATPase reaction velocity was even increased at higher ATP contents, whereas a marked decrease in ATPase reaction velocity was found if ATP levels decreased. The equilibrium of the CK-reaction remained unchanged following CP, whereas IP induced a changing CK equilibrium, which was the more shifted towards PCr the more myocardial HEP content decreased. The data demonstrate different effects of CP and IP on myocardial HEP metabolism, i.e. PCr and ATP breakdown as well as the apparent equilibrium of the creatine kinase (CK)-reaction. For these reasons the combination of the two protective interventions has an additive effect.


Mobile elements in a single-filament orange Guaymas Basin Beggiatoa ("Candidatus Maribeggiatoa") sp. draft genome: evidence for genetic exchange with cyanobacteria.
The draft genome sequence of a single orange Beggiatoa ("Candidatus Maribeggiatoa") filament collected from a microbial mat at a hydrothermal site in Guaymas Basin (Gulf of California, Mexico) shows evidence of extensive genetic exchange with cyanobacteria, in particular for sensory and signal transduction genes. A putative homing endonuclease gene and group I intron within the 23S rRNA gene; several group II catalytic introns; GyrB and DnaE inteins, also encoding homing endonucleases; multiple copies of sequences similar to the fdxN excision elements XisH and XisI (required for heterocyst differentiation in some cyanobacteria); and multiple sequences related to an open reading frame (ORF) (00024_0693) of unknown function all have close non-Beggiatoaceae matches with cyanobacterial sequences. Sequences similar to the uncharacterized ORF and Xis elements are found in other Beggiatoaceae genomes, a variety of cyanobacteria, and a few phylogenetically dispersed pleiomorphic or filamentous bacteria. We speculate that elements shared among filamentous bacterial species may have been exchanged in microbial mats and that some of them may be involved in cell differentiation.


Anaesthetic experience in female sterilisation at Jos University Teaching Hospital, Nigeria.
To present the trend of anaesthetics used for interval and post-partum female sterilisation. A retrospective descriptive study. Jos University Teaching Hospital, Jos, Nigeria from 1985 to 2004. All cases of female sterilisation in Jos University Teaching Hospital within the study period. Clients were allowed free choice of the method of anaesthesia after appropriate counseling. Written consent by the patient and her husband was obtained. During the period, 4,313 female sterilisations were performed. The mean (+/-SD) age and parity of the clients were 36.8 +/- 4.8 years, and 7.6 +/- 2.4 respectively. Local anaesthesia alone was the most commonly used (75.0%), followed by general anaesthesia (15.8%) and local anaesthesia with sedation (9.2%). The use of local anaesthesia alone for minilaparotomy under local anaesthesia for female sterilisation rose from 0.0% in 1985 to 83.0% in 2004. Majority (79.1%) of the cases were performed as interval procedures, 15.1% performed at Caesarean section and 5.8% as postpartum procedures. There were no serious morbidity and/or mortality associated with the types of anaesthesia used. Minilaparotomy under local anaesthesia for sterilisation has been found to be feasible, and now acceptable in our institution.


The anatomic relationship of the esophageal lead to the left atrium.
This study was undertaken to establish the "in vivo" anatomic relationship of the esophageal catheter to the left atrium. Fourteen patients, seven of whom had enlarged left atria, were included in the study. During cardiac catheterization, a bipolar recording catheter was placed in the esophagus; left atrial cineangiograms were then obtained in the posterior-anterior (PA), left anterior oblique (LAO), and lateral views. The distance of the esophageal catheter, in each of four commonly used positions, to the interatrial septum-right atrium was measured in the PA and LAO cineangiograms. The distance from the same catheter positions to the posterior left atrial wall was measured from the lateral view. In all positions and regardless of left atrial size, the catheter clearly lay behind the left atrium and was closer to the left atrial wall than any other cardiac structure, including the interatrial septum. Bipolar electrograms were recorded from the four commonly used esophageal locations; atrial wave amplitude and other parameters of the electrogram deflection did not vary significantly in the different lead positions. We conclude that esophageal electrograms reflect left atrial activity by virtue of the recording electrode's clear proximity to this structure. The amplitude and configuration of the atrial wave itself is generally not useful in determining the position of the esophageal lead.


[Renal size in children with normal renal 99mTc-DMSA scan: building a nomogram with scintigraphic length].
The aim of this study was to evaluate renal size in children with normal 99mTc-DMSA scans and to compare it with the published ultrasonographic values. Renal length of DMSA normal scans obtained from 253 children with history of urinary tract infection was measured. Regression analysis and length/age curves were done. Scintigraphic values were compared with published normal ultrasonographic length. Regression algorithm for the whole group of patients was length (cm) = 6.1727 age (yr)0,1535. In the first year of life the equation was length (cm) = 0.1524 age (month) + 4.568, and in 1-16 yr children it was length (cm) = 0.2644 age (yr) + 6.319. There were no significant differences between both genders or in regards to reference ultrasonographic data. Both renal 99mTc-DMSA scintigraphy and ultrasonography are useful tools for estimating renal size in children according age.


Store-operated calcium entry modulates neuronal network activity in a model of chronic epilepsy.
Store-operated Ca(2+) entry (SOCE) over the plasma membrane is activated by depletion of intracellular Ca(2+) stores and has only recently been shown to play a role in CNS processes like synaptic plasticity. However, the direct effect of SOCE on the excitability of neuronal networks in vitro and in vivo has never been determined. We confirmed the presence of SOCE and the expression of the calcium sensors STIM1 and STIM2, which convey information about the calcium load of the stores to channel proteins at the plasma membrane, in neurons and astrocytes. Inhibition of SOCE by pharmacological agents 2-APB and ML-9 reduced the steady-state neuronal Ca(2+) concentration, reduced network activity, and increased synchrony of primary neuronal cultures grown on multi-electrode arrays, which prompted us to elucidate the relative expression of STIM proteins in conditions of pathologic excitability. Both proteins were increased in brains of chronic epileptic rodents and strongly expressed in hippocampal specimens from medial temporal lobe epilepsy patients. Pharmacologic inhibition of SOCE in chronic epileptic hippocampal slices suppressed interictal spikes and rhythmized epileptic burst activity. Our results indicate that SOCE modulates the activity of neuronal networks in vitro and in vivo and delineates SOCE as a potential drug target.


A case of acrodermatitis enteropathica.
Acrodermatitis enteropathica is a rare hereditary disorder affecting zinc metabolism that is characterized by dermatitis, alopecia, gastrointestinal disturbances, eye infections, and growth failure. We report a 17-month-old girl with acrodermatitis enteropathica. Physical examination showed a cutaneous eruption consisting of vesiculobullous and psoriasiform skin lesions symmetrically distributed in the perioral, acral, and perineal areas. Her plasma zinc level was decreased (75 micrograms/dl), but within the normal range (60.00-135.00 micrograms/dl). The patient was given zinc sulfate 50 mg/day. At the end of two months, she had significantly improved.


Relationships between tibial rim alignment and joint space width measurement reproducibility in non-fluoroscopic radiographs of osteoarthritic knees.
Superimposition of the rims of the medial tibial plateau to within 1mm is an aim of fluoroscopic knee positioning protocols for osteoarthritic (OA) knee radiography and has also been proposed as a measure of quality for non-fluoroscopic methods. To evaluate the effect of tibial rim alignment (TRA) on reproducibility of joint space width (JSW) measurement, both were measured from radiographs taken with each tibial plateau at a range of angles determined by different non-fluoroscopic views. TRA and JSW measurements were taken from both knees of 100 OA patients each radiographed in fully extended, schuss/tunnel, and MTP views. Degree of TRA was compared with JSW reproducibility using correlation, and between groups defined both by the 1mm threshold and by TRA-defined quartiles. JSW reproducibility was dependent on the degree of TRA in the fully extended and schuss/tunnel flexed knee views, although the use of the specific TRA threshold of 1mm was not supported. In the MTP view, JSW measurement was found to be highly reproducible across the full range of TRA values. These results contradict claims that TRA to within 1mm is essential for useful measurement of JSW. It is an arbitrary threshold, of use in quality control (QC) for protocols which explicitly require such alignment, and the choice of QC criteria for other protocols should be evaluated on a view-by-view basis. The results confirm previous studies showing the MTP view to afford highly reproducible JSW measurement.


Epidemiological study of scabies in different environmental conditions in central poland.
The goal of the research was to determine the spreading of Sarcoptes scabiei and the incidence of scabies in the residents of particular districts of the Swietokrzyskie Voivodeship (Central Europe) from 1990 to 1998. An epidemiological analysis of scabies incidents was performed, with a focus on the age and sex patterns of the population, seasonal incidence, and environmental conditions. In the entire area covered in the period studied, a total of 2064 cases of scabies were reported. The incidence of scabies was typically higher in rural areas than in cities. The most cases of scabies were noted in children and teenagers between 6 and 15 years of age. The research indicated that prevalence of the disease within a family is an important factor in scabies epidemiology in the Swietokrzyskie Voivodeship. The incidence of scabies is seasonal in its nature, as the majority of cases occurred in the autumn and winter months. The incidence of the disease can be reduced by improving socioeconomic and hygienic conditions and by implementing a proper system of social education, as well as by promoting more efficient health service.


Establishment of an interleukin-5-dependent subclone from an interleukin-3-dependent murine hemopoietic progenitor cell line, LyD9, and its malignant transformation by autocrine secretion of interleukin-5.
An interleukin-5 (IL-5)-dependent subclone, K-5, was established from an IL-3-dependent murine hemopoietic progenitor cell line by co-culturing with bone marrow stroma cells. K-5 cells were induced to differentiate into myeloid lineage cells by co-culturing with cloned PA6 stroma cells. By co-culturing with another cloned stroma cell (ST-2s10), K-5 cells gave rise to a factor-independent transformant cell line LT-5 which proliferated in an autocrine manner by secretion of IL-5 and produced tumors in nude mice. Molecular cloning of the IL-5 gene of LT-5 cells and the nucleotide sequencing of its 5' flanking region indicate that a transposition of an intracisternal A-particle (IAP) element to the 5' flanking region of the IL-5 gene is responsible for the constitutive expression of IL-5 mRNA of an aberrant size in LT-5 cells.


Generation of Human Neural Stem Cells by Direct Phenotypic Conversion.
Human neural stem cells (hNSC) are multipotent adult stem cells. Various studies are underway worldwide to identify new methods for treatment of neurological diseases using hNSC. This chapter summarizes the latest research trends in and fields for application of patient-specific hNSC using direct phenotypic conversion technology. The aim of the study was to analyze the advantages and disadvantages of current technology and to suggest relevant directions for future hNSC research.


Obesity as a comorbidity in children and adolescents with autoimmune rheumatic diseases.
Childhood obesity is the public health issue with alarming rates recorded throughout developed world and an important modifiable health risk for developing various chronic diseases, with childhood-onset autoimmune rheumatic diseases among them also. The aim of this article was to summarize epidemiological, pathophysiological and clinical implication of obesity on juvenile idiopathic arthritis (JIA), childhood-onset systemic lupus erythematosus (cSLE), juvenile dermatomyositis (JDM), IgA vasculitis (IgAV) and Kawasaki disease (KD). We reviewed PubMed database and selected 74 relevant articles. Epidemiological data of obesity among children with autoimmune rheumatic diseases indicate an increased prevalence of it. Pathophysiological link between obesity, humoral adipokines and cytokines released from fat tissue and childhood-onset autoimmune rheumatic diseases is complex and still not entirely clear. From the clinical point of view, obesity was not associated with disease activity in JIA and cSLE, but proved to contribute on functional impairment in both diseases and affect poor treatment response in JIA patients. Early atherosclerosis and cardiovascular disease (CVD) development in obese children and adolescents with JIA, cSLE and JDM are certainly important obesity-related complications. Understanding how obesity affects children and adolescents with autoimmune rheumatic diseases may encourage clinicians to consider taking better preventive strategies in this population to improve their long-term outcome.


User Experiences in a Digital Intervention to Support Total Skin Self-examination by Melanoma Survivors: Nested Qualitative Evaluation Embedded in a Randomized Controlled Trial.
Melanoma is a relatively common cancer type with a high survival rate, but survivors risk recurrences or second primaries. Consequently, patients receive regular hospital follow-up, but this can be burdensome to attend and not optimally timed to detect arising problems. Total skin self-examination (TSSE) supports improved clinical outcomes from melanoma via earlier detection of recurrences and second primaries, and digital technology has the potential to support TSSE. Recent research with app-based interventions aimed at improving the well-being of older adults has found that they can use the technology and benefit from it, supporting the use of digital health care in diverse demographic groups. Thus, the Achieving Self-directed Integrated Cancer Aftercare (ASICA) digital health care intervention was developed. The intervention provided melanoma survivors with a monthly prompt to perform a TSSE as well as access to a dermatology nurse who provided them with feedback on photographs and descriptions of their skin. We aimed to explore participants' attitudes, beliefs, and experiences regarding TSSE practices. Furthermore, we explored how participants experienced technology and how it influenced their practice of TSSE. Finally, we explored the practical and technical experiences of ASICA users. This was a nested qualitative evaluation within a dual-center randomized controlled trial of the ASICA intervention. We conducted semistructured telephone interviews with the participants during a randomized controlled trial. The participants were purposively sampled to achieve a representative sample with representative proportions by age, sex, and residential geography. Interviews were transcribed verbatim and analyzed using a framework analysis approach applied within NVivo 12. A total of 22 interviews were conducted with participants from both groups. In total, 40% (9/22) of the interviewed participants were from rural areas, and 60% (13/22) were from urban areas; 60% (13/22) were from the intervention group, and 40% (9/22) were from the control group. Themes evolved around skin-checking behavior, other people's input into skin checking, contribution of health care professionals outside ASICA and its value, ideas around technology, practical experiences, and potential improvements. ASICA appeared to change participants' perceptions of skin checking. Users were more likely to report routinely performing TSSE thoroughly. There was some variation in beliefs about skin checking and using technology for health care. Overall, ASICA was experienced positively by participants. Several practical suggestions were made for the improvement of ASICA. The ASICA intervention appeared to have positively influenced the attitudes and TSSE practices of melanoma survivors. This study provides important qualitative information about how a digital health care intervention is an effective means of prompting, recording, and responding to structured TSSE by melanoma survivors. Technical improvements are required, but the app offers promise for technologically enhanced melanoma follow-up in future. ClinicalTrials.gov NCT03328247; https://clinicaltrials.gov/ct2/show/NCT03328247?term=ASICA&rank=1. RR2-10.1186/s13063-019-3453-x.


The role of memory and language ability in children's production of two-clause sentences containing before and after.
We assessed 3- to 6-year-old children's production of two-clause sentences linked by before or after. In two experiments, children viewed an animated sequence of two actions and were asked to describe the order of events in specific target sentence structures. We manipulated whether the target sentence structure matched the chronological order of events (e.g., "He finished his homework, before he played in the garden" [chronological order]) or not (e.g., "Before he played in the garden, he finished his homework" [reverse order]). Children produced fewer accurate target sentences when the presentation order of the two clauses did not match the chronological order of events, specifically for target sentences linked by after. Independent measures of vocabulary and memory both were related to performance, but vocabulary was the stronger predictor. We conclude that developmental improvements in children's ability to produce two-clause sentences linked by a sequential temporal connective are driven primarily by language ability rather than memory capacity per se. This work also highlights the advantages of using both sentence repetition (Experiment 1) and blocked elicited production (Experiment 2) paradigms to elicit sentence production in young children.


Oxidative-stress and long-term hepatotoxicity: comparative study in Upcyte human hepatocytes and hepaRG cells.
Drug-induced liver injury (DILI) is one of the most common and serious adverse drug reactions and a major cause of drug development failure and withdrawal. Although different molecular mechanisms are implicated in DILI, enhanced ROS levels have been described as a major mechanism. Human-derived cell models are increasingly used in preclinical safety assessment because they provide quick and relatively inexpensive information in early stages of drug development. We have analyzed and compared the phenotype and functionality of two liver cell models (Upcyte human hepatocytes and HepaRG cells) to demonstrate their suitability for long-term hepatotoxicity assessments and mechanistic studies. The transcriptomic and functional analysis revealed the maintenance of phase I and phase II enzymes, and antioxidant enzymes along time in culture, although the differences found between both test systems underlie the differential sensitivity to hepatotoxins. The evaluation of several mechanisms of cell toxicity, including oxidative stress, by high-content screening, demonstrated that, by combining the stable phenotype of liver cells and repeated-dose exposure regimes to 12 test compounds at clinically relevant concentrations, both Upcyte hepatocytes and HepaRG offer suitable properties to be used in routine screening assays for toxicological assessments during drug preclinical testing.


The biochemical modes of action of vitamin e and selenium: A hypothesis.
In this paper recent experimental work on rat liver is discussed which is considered to indicate that a primary function of vitamin E in vivo may be to inhibit the oxidation of selenide-containing proteins present in mitochondria and in smooth endoplasmic reticulum. On the basis of molecular model building studies, it is also suggested that the well-known nutritional relationships between vitamin E and dietary polyunsaturated lipids may be due to the occurrence within normal membranes of specific complexes between the vitamin and some of the molecules of polyunsaturated phospholipids. Without the vitamin, membranes may have an abnormally high permeability and they may be subject to degradation by endogenous phospholipases in vivo, as well as being abnormally susceptible to damage in vitro by dialuric acid and hydrogen peroxide.