A reappraisal of the feedback effects of oestradiol upon luteinizing hormone surge.
The objective of this study was to assess the temporal relationships of serum oestradiol and luteinizing hormone (LH) with regard to the feedback mechanism leading to the LH surge. Daily measurements of oestradiol, LH and follicle stimulating hormone were made in 65 women with ovarian failure undergoing an evaluation of endometrial response to oral cycle stimulation. Patients received incremental oral oestradiol valerate for varying durations at the in-vitro fertilization unit of the Sheba Medical Center, Tel Hashomer, Israel. The treatment protocol involved the administration of daily oestradiol valerate, starting with 1 mg/day, followed by a daily increment of 1 mg for a total of 4, 6 and 8 days. After the last day, a daily maintenance dose of 2 mg was continued. Serum oestradiol concentrations correlated with the dose of oral oestradiol valerate. Peak oestradiol concentrations (mean +/- SEM) were 572 +/- 60, 721 +/- 42 and 797 +/- 53 pg/ml for 4, 6 and 8 days of oestradiol valerate administration respectively. Serum LH, after an initial suppression, peaked 2 days after maximal oestradiol valerate dose and 1 day after peak serum oestradiol. The magnitude of the LH peak was proportional to the duration of incremental oestradiol valerate treatment. In conclusion, the LH surge is temporally related to the cessation of serum oestradiol increase. Consequently, the occurrence of this surge can be explained by a simple negative feedback inhibition of pituitary LH release rather than by a dual negative/positive mechanism.


Determination of sugars, starches, and total dietary fiber in selected high-consumption foods.
A general scheme has been developed to determine sugars, starches, and total dietary fiber (TDF) in half-gram freeze-dried subsamples of various foods (or wet samples containing about 0.5 g dry matter). Duplicate subsamples are extracted for free sugars with 80% methanol, dried, derivatized to their trimethylsilylated oximes or ethers, and quantitated by gas-liquid chromatography (GLC). Residues after 80% methanol extraction are incubated with a solution containing amyloglucosidase in acetate buffer. Hydrolyzates are centrifuged, and duplicate aliquots are removed for glucose determination by GLC. Starch content is calculated as glucose (g/100 g) x 0.9. Remaining hydrolyzates are diluted with 4 volumes of 95% ethanol, left at room temperature for 1 h, and then filtered through glass crucibles matted with Celite filter aid. The weight of dry residues are corrected for residual crude protein and ash, and the resulting values are taken to be the TDF content of a sample. A variety of high-consumption foods selected by the Nutrient Data Laboratory of the Agricultural Research Service were analyzed for carbohydrate fractions by this method. Values for total sugar and dietary fiber were compared with those obtained by a commercial laboratory using different methods.


Site-directed mutagenesis of glutamate residues in the large extrinsic loop of the photosystem II protein CP 43 affects oxygen-evolving activity and PS II assembly.
The psbC gene encodes the intrinsic chlorophyll protein CP 43, a component of photosystem II in higher plants, green algae, and cyanobacteria. Oligonucleotide-directed mutagenesis was used to introduce mutations into the portion of psbC that encodes the large extrinsic loop E of CP 43 in the cyanobacterium Synechocystis 6803. Three mutations, E293Q, E339Q, and E352Q, each produced a strain with impaired photosystem II activity. The E293Q mutant strain grew photoautotrophically at rates comparable to the control strain. Immunological analyses of several PS II components indicated that this mutant accumulated normal quantities of PS II proteins. However, this mutant evolved oxygen to only 56% of control rates at saturating light intensities. Measurements of total variable fluorescence yield indicated that this mutant assembled approximately 60% of the fully functional PS II centers found in the control strain. The E339Q mutant grew photoautotrophically at a severely reduced rate. Both immunological analysis and variable fluorescence yield experiments indicated that E339Q assembled a normal complement of PS II centers. However, this mutant was capable of evolving oxygen to only 20% of control rates. Variable fluorescence yield experiments demonstrated that this mutant was inefficient at using water as an electron donor. Both E293Q and E339Q strains exhibited an increased (approximately 2-fold) sensitivity to photoinactivation. The E352Q mutant was the most severely affected. This mutant failed to grow photoautotrophically and exhibited essentially no capacity for oxygen evolution. Measurements of total variable fluorescence yield indicated that this mutant assembled no functional PS II centers. Immunological analysis of isolated thylakoid membranes from E352Q revealed a complete absence of CP 43 and reduced levels of both the D1 and manganese-stabilizing proteins. These results suggest that the mutations E293Q and E339Q each produce a defect associated with the oxygen-evolving complex of photosystem II. The E352Q mutation appears to affect the stability of the PS II complex. This is the first report showing that alteration of negatively charged residues in the CP 43 large extrinsic loop results in mutations affecting PS II assembly/function.


Hepatitis B virus genotypes among chronic hepatitis B patients reporting at Korle-Bu teaching hospital, Accra, Ghana.
Knowledge of hepatitis B virus (HBV) genotype is an important predictive variable which might have an impact in management and treatment of patients with chronic hepatitis B infection. In Ghana very little information is available on hepatitis B genotypes. This study was conducted to determine the distribution of HBV genotypes circulating among chronic hepatitis B patients reporting at the Korle-Bu Teaching Hospital (KBTH), Accra, Ghana. Blood samples (10 ml) were collected from 250 consenting patients. DNA was extracted and amplified using polymerase chain reaction technique. Restriction fragment length polymorphism (RFLP) was used for the detection of genotypes. Out of the 250 chronic hepatitis B patients who were HBsAg positive, 91 (36.4%) were males aged 29.8 ± 9.1 and 159 (63.6%) females aged 33± 12.1 years. HBV DNA was detected in 111 (44.4%) but only 58 (52%) of these were typeable. These were classified as genotype A, 8 (7.2%); genotype D, 3 (2.7%) and genotype E, 47 (42.3%). Our results did not show any association between the infecting genotype and age (X2= 0.923; p-value=0.623) or gender (X2= 0.283, p= 0.579). Consistent with similar studies worldwide, the results suggest that genotypes A, D and E were the genotypes circulating among chronic hepatitis B patients who reported to the Korle-Bu Teaching Hospital with genotype E being the most predominant and therefore constitutes an important public health concern. We recommend further epidemiological studies to understand the implication of genotype E in terms of disease progression and treatment.


Distal clavicle osteolysis following fixation with a synthetic ligament.
We present a case of distal clavicle osteolysis following treatment of a chronic acromioclavicular joint dislocation with a synthetic ligament. The relevant literature is reviewed and discussed.


Identification of carbonic anhydrase XII as the membrane isozyme expressed in the normal human endometrial epithelium.
Although previous studies demonstrated carbonic anhydrase (CA) activity in the human endometrium, the CA isozyme(s) responsible for this activity has not been established. In this report, we provide the first evidence that the CA isozyme XII, a recently identified transmembrane isozyme that is expressed in normal kidney and greatly overexpressed in some renal cancers, is present in endometrium. We show by immunohistochemistry that CA XII is expressed in the basolateral plasma membrane of epithelial cells of normal human endometrium. Expression of CA XII in uterus was confirmed by Northern blotting. Detergent-solubilized CA XII was isolated from human endometrium by inhibitor affinity chromatography and characterized by isoelectric focusing and Western blot as a polypeptide with a pI of 6.3. The high expression of CA XII in the endometrial epithelium suggests that it may be functionally linked to the pH-dependent events in spermatozoa that precede fertilization. Its basolateral location and extracellular active site could also allow it to influence the morphological changes in endometrium that occur during the menstrual cycle.


The Bacterial Community Structure and Microbial Activity in a Traditional Organic Milpa Farming System Under Different Soil Moisture Conditions.
Agricultural practices affect the bacterial community structure, but how they determine the response of the bacterial community to drought, is still largely unknown. Conventional cultivated soil, i.e., inorganic fertilization, tillage, crop residue removal and maize (Zea mays L.) monoculture, and traditional organic farmed soil "milpa," i.e., minimum tillage, rotation of maize, pumpkin (Cucurbita sp.) and beans (Phaseolus vulgaris L.) and organic fertilization were sampled. Both soils from the central highlands of Mexico were characterized and incubated aerobically at 5% field capacity (5%FC) and 100% field capacity (FC) for 45 days, while the C and N mineralization, enzyme activity and the bacterial community structure were monitored. After applying the different agricultural practices 3 years, the organic C content was 1.8-times larger in the milpa than in the conventional cultivated soil, the microbial biomass C 1.3-times, and C and N mineralization 2.0-times (mean for soil incubated at 5%FC and FC). The dehydrogenase, activity was significantly higher in the conventional cultivated soil than in the milpa soil when incubated at 5%FC, but not when incubated at FC. The relative abundance of Gemmatimonadetes was larger in the conventional cultivated soil than in the milpa soil in soil both at 5%FC and FC, while that of Bacteroidetes showed an opposite trend. The relative abundance of other groups, such as Nitrospirae and Proteobacteria, was affected by cultivation technique, but controlled by soil water content. The relative abundance of other groups, e.g., FBP, Gemmatimonadetes and Proteobacteria, was affected by water content, but the effect depended on agricultural practice. For soil incubated at FC, the xenobiotics biodegradation and metabolism related functions were higher in the milpa soil than in the conventional cultivated soil, and carbohydrate metabolism showed an opposite trend. It was found that agricultural practices and soil water content had a strong effect on soil characteristics, C and N mineralization, enzyme activity, and the bacterial community structure and its functionality. Decreases or increases in the relative abundance of bacterial groups when the soil water content decreased, i.e., from FC to 5%FC, was defined often by the cultivation technique, and the larger organic matter content in the milpa soil did not prevent large changes in the bacterial community structure when the soil was dried.


Functional analysis of a reproductive organ predominant expressing promoter in cotton plants.
Transgenic Bt insect-resistant cotton plants have high insect resistance in the early stage of development, but relatively low resistance in the late stage. Substituting a reproductive organ-specific promoter for the CaMV35S promoter presently being used could be an ideal solution. For the first time, the promoter sequence of ADP-ribosylation factor 1 (arf1) gene was isolated from Gossypium hirsutumY18 by means of inverse PCR. The sequencing result discovered the unique structure of the arf1 promoter, including four promoter-specific elements, the initiator, TATA box, CAAT box and GC box, and also an intron in 5'-untranslation region. Four plant expression vectors were constructed for functional analysis of the promoter. Based on the pBl121 plant expression vector, four truncated arf1 promoters took the place of the CaMV35S promoter. These vectors were different only in their promoter regions. They were introduced into cotton plants via pollen tube pathway. Histochemical GUS staining and fluorescence quantitative analyses were performed to examine the expression patterns of the GUS gene driven by the 4 arf1 truncated promoters in transgenic cotton plants respectively. The results showed that the arf1 promoter was a typical reproductive organ-specific promoter. Hopefully, the arf1 promoter can be a regulatory element for designing cotton reproductive organs with desired characteristics.


Regulation of c-fos expression by static stretch in rat myometrial smooth muscle cells.
We investigated whether mechanical stretch stimulates expression of the immediate early gene c-fos messenger RNA in rat myometrial smooth muscle cells. Freshly isolated myometrial smooth muscle cells (from virgin rats) were plated on flexible culture plates that were coated with either collagen I, laminin, elastin, or pronectin and were subjected to a static stretch with a computerized strain unit. Immunocytochemistry was used to define smooth muscle cell phenotype, and c-fos expression levels were assessed by Northern blotting. Myometrial smooth muscle cells maintained their phenotype for at least 7 days in culture. Stretch induced a dramatic increase in levels of c-fos messenger RNA, which peaked after 30 minutes. Stimulation of c-fos gene expression was dependent on the degree of stretch (maximum induction at 25% elongation). Interestingly, as little as 1 minute of stretch was sufficient to induce maximal c-fos expression at 30 minutes. The most dramatic induction of c-fos was observed in smooth muscle cells that were stretched on collagen I-coated plates. Myometrial smooth muscle cells can directly respond to mechanical stretch with a strength- and matrix component-dependent increase in c-fos messenger RNA. The time delay between the initial stimulus and the cumulative elevation of c-fos messenger RNA suggests a complex regulation from mechanoreception to c-fos gene expression.


Oral immunoglobulin for the treatment of rotavirus diarrhea in low birth weight infants.
Rotavirus infection is the most common neonatal nosocomial viral infection. It is a major health problem worldwide. Epidemics with the newer P(6)G9 strains have been reported in neonatal units globally. These strains can cause severe symptoms in most infected infants. Infection control measures become necessary and the utilization of hospital resources increase. Local mucosal immunity in the intestine to rotavirus is important in the resolution of infection and protection against subsequent infections. Boosting local immunity by oral administration of anti-rotaviral immunoglobulin preparations might be a useful strategy in treating rotaviral infections, especially in low birth weight babies. To determine the effectiveness and safety of oral immunoglobulin preparations for the treatment of rotavirus diarrhea in hospitalized low birth weight infants (birth weight less than 2500 g) Electronic databases including The Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 3, 2004), MEDLINE, EMBASE and CINAHL, Biological Abstracts (BIOSIS) were searched by the strategy outlined in the protocol. Science Citation Index search for all articles that referenced Barnes 1982 were searched. The proceedings of the Pediatric Academic Societies published online at 'Abstracts Online' were searched. Ongoing registered trials at www.clinicaltrials.gov and www.controlled-trials.com were searched. Authors prominent in the field were contacted for any unpublished articles and more information on published articles was sought. Reference lists of identified clinical trials and personal files were also reviewed. The above search was updated in July 2011. The criteria used to select studies for inclusion were: 1) DESIGN: randomized or quasi-randomized controlled trials 2) Hospitalized low birth weight infants with rotavirus diarrhea 3) INTERVENTION: Oral immunoglobulin preparations compared to placebo or no intervention 4) At least one of the following outcomes were reported: All cause mortality during hospital stay, mortality due to rotavirus infection during hospital stay, duration of diarrhea, need for rehydration, duration of viral excretion, duration of infection control measures, length of hospital stay in days, recurrent diarrhea or chronic diarrhea The two reviewers were to independently abstract data from eligible trials. No data were available for analysis. No eligible randomized controlled trials were found. No randomized controlled trials that assessed the effectiveness or safety of oral immunoglobulin preparations for the treatment of rotavirus diarrhea in hospitalized low birth weight infants were found. Clinical trials that address the issue of oral immunoglobulin treatment of rotavirus infection are needed.


Analysis of green kiwi fruit (Actinidia deliciosa cv. Hayward) proteinases by two-dimensional zymography and direct identification of zymographic spots by mass spectrometry.
Proteinases present in kiwi fruits are potentially allergenic enzymes belonging to the papain family of cysteine proteinases. Actinidin is a prominent kiwi enzyme. The study of kiwi proteinases is important for the follow-up of fruit maturation, a deeper insight in the allergenic properties of individual proteins, and the application of kiwi proteinases for meat tenderisation and other industrial purposes. Kiwi crude extracts were analysed by two-dimensional zymography on gelatin-containing gels. The digestion by the reactivated proteolytic enzymes after electrophoresis resulted in insights into kiwi proteinases. A mixture of several enzyme isotypes with the same pI but different molecular mass was observed. Clear spots, corresponding to the proteolytic activities, were excised, digested with trypsin, and submitted to MALDI-ToF mass spectrometry for protein identification. The most representative enzyme was actinidin. The innovative achievements of the present study are the: (1) two-dimensional zymographic map of kiwi gelatinases without the need for extensive purification; and (2) direct identification of proteinase isotypes by means of direct MALDI-ToF MS analysis of the zymographic spots.


Diagnostic Difficulties in Discriminating Autism Spectrum Disorder in an Adult With Periodic Psychosis Versus Schizophrenia Spectrum Condition: An Insight From Psychological Testing.
It has long been recognized that the biological underpinnings of autism spectrum disorder (ASD) and schizophrenia spectrum disorder (SSD) may share a common basis; however, the two conditions remain separate in the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-5) due to a few distinguishing characteristics. Both disorders are characterized by cognitive and social deficits and have been presumed to be linked to multiple genes. We describe a 46-year-old male who presented atypically with three previous and one current episode of schizoaffective-like symptoms. We describe his previous inpatient admissions, current inpatient course, psychological test results, and treatment. The patient initially presented with schizoaffective disorder, but with a thorough interview, collateral information review, and psychological evaluation, it was determined that he instead was presenting with a previously undiagnosed case of ASD with brief psychosis when under stress. This case serves as an example of an atypical presentation of ASD which can be mistaken for schizoaffective disorder. It is important to establish the correct diagnosis, as the subsequent treatment and management of the patient's problems will depend on it. In such a patient, a low dose of atypical antipsychotic medication with serotonergic properties and psychotherapy would be the treatment of choice.


Interferon-induced transcription of a gene encoding a 15-kDa protein depends on an upstream enhancer element.
A human gene encoding an interferon-induced 15-kDa protein has been isolated from a genomic library. The gene appears to be single-copy and is composed of two exons, the first of which contains the ATG translation initiation codon. In vitro nuclear run-on assays showed that the transcription rate of the gene is stimulated after interferon treatment. To analyze transcriptional regulatory sequences, we constructed recombinant plasmids for use in transient transfection assays of HeLa cells. Constructs containing 115 nucleotides 5' to the transcription initiation site were found to be fully inducible by interferon. Assays of deletion mutants identified a critical element for interferon induction located between -115 and -96, just upstream of the "CCAAT box." Moreover, a DNA fragment including this region can confer interferon inducibility on a heterologous promoter (thymidine kinase) when cloned in either orientation upstream of the gene or downstream of the gene. These are properties characteristic of an enhancer element that is active only after treatment with interferon. This regulatory sequence may be shared by a group of interferon-induced genes, since a very similar sequence is present within the functional region near the RNA start site of another interferon-induced gene.


Politics, technology, and transformation: a conversation with Newt Gingrich.
In this interview conducted by Jeff Goldsmith, former congressman and Speaker of the House Newt Gingrich (R-GA) provides an iconoclastic view of the problem of reforming a Medicare system in the absence of a political consensus about what needs reforming. Topics include Medicare prescription drug legislation, the role of technology in transforming the health care system, the need to communicate clearly and honestly with people (especially senior citizens) about proposed changes to their health benefits, the role of the private sector, and incentives for change.


Vector analysis of the wall shear rate at the human aortoiliac bifurcation using cine MR velocity mapping.
Small or oscillatory wall shear stress accelerates atherosclerosis. MR velocity mapping is feasible for vector analysis of wall shear rate (a spatial gradient of blood flow velocity at the vessel wall) in humans. A relationship between anatomic variations at the aortoiliac bifurcation and characteristics of wall shear rate was evaluated. To obtain two components of wall shear rate vectors, an axial component along the vessel axis and a nonaxial component perpendicular to the former at the inner and outer walls of the common iliac arteries just distal to the aortoiliac bifurcation, we performed cine MR velocity mapping with three orthogonal velocity-encoded directions in seven volunteers. The peak axial component at the outer wall (120.6 +/- 37.2 sec(-1)) was smaller than that at the inner wall (196.0 +/- 53.7 sec(-1)) (p < 0.01). Oscillation described by a time integral of the axial component in recessive blood flow direction over integrals in dominant and recessive directions at the outer wall was greater (0.24 +/- 0.11) than that at the inner wall (0.15 +/- 0.08) (p < 0.01). The intersecting angle between the extrapolation of the aortic axis and the direction of the axis of the common iliac artery correlated positively with the peak axial component (r = 0.577, p < 0.05) and inversely with oscillation (r = 0.603, p < 0.05). Three-dimensional vector analysis with MR velocity mapping revealed that the outer wall at the aortoiliac bifurcation showed low and oscillatory shear rate, and this inclination was increased when the takeoff angle of the iliac artery was small.


Cognition, emotion, and attention.
Deficits of attention, emotion, and cognition occur in individuals with alcohol abuse and addiction. This review elucidates the concepts of attention, emotion, and cognition and references research on the underlying neural networks and their compromise in alcohol use disorder. Neuroimaging research on adolescents with family history of alcoholism contributes to the understanding of pre-existing brain structural conditions and characterization of cognition and attention processes in high-risk individuals. Attention and cognition interact with other brain functions, including perceptual selection, salience, emotion, reward, and memory, through interconnected neural networks. Recent research reports compromised microstructural and functional network connectivity in alcoholism, which can have an effect on the dynamic tuning between brain systems, e.g., the frontally based executive control system, the limbic emotion system, and the midbrain-striatal reward system, thereby impeding cognitive flexibility and behavioral adaptation to changing environments. Finally, we introduce concepts of functional compensation, the capacity to generate attentional resources for performance enhancement, and brain structure recovery with abstinence. An understanding of the neural mechanisms of attention, emotion, and cognition will likely provide the basis for better treatment strategies for developing skills that enhance alcoholism therapy adherence and quality of life, and reduce the propensity for relapse.


Snore sound recognition: On wavelets and classifiers from deep nets to kernels.
In this paper, we present a comprehensive comparison of wavelet features for the classification of snore sounds. Wavelet features have proven to be efficient in our previous work; however, the benefits of wavelet transform energy (WTE) and wavelet packet transform energy (WPTE) features were not clearly established. In this study, we firstly present our updated snore sounds database, expanded from 24 patients (collected by one medical centre) to 40 patients (collected by three medical centres). We then study the effects of varying frame sizes and overlaps for extraction of the wavelet low-level descriptors, the effect of which have yet to be fully established. We also compare the performance of the WTE and WPTE features when fed into multiple classifiers, namely, Support Vector Machines (SVM), K-Nearest Neighbours, Linear Discriminant Analysis, Random Forests, Extreme Learning Machines, Kernel Extreme Learning Machines, Multilayer Perceptron, and Deep Neural Networks. Key results presented indicate that, when fed into a SVM, WTE outperforms WPTE (one-tailed z-test, p<;0.002). Further, WPTE can achieve a significant improvement when trained by a k-nearest neighbours classifier (one-tailed z-test, p <; 0.001).


Comparative studies of Ca N-acetylhomotaurinate and Ca N-acetyltaurinate. I. Effects on the ionic transfer through the isolated human amnion.
The effects of two taurine and homotaurine derivatives (Ca N-acetyltaurinate: ATACa and Ca N-acetylhomotaurinate: AOTACa) were studied on the ionic transfer through a membranous model: the human isolated amniotic membrane. The ionic transfer was evaluated by measurements of the various components (cellular and paracellular) of the total conductance across the membrane. AOTACa influenced all the components of the total ionic conductance without flux ratio modifications and the general action scheme was a biphasic effect: decrease conductance with low concentrations and increase conductance with high concentrations. This effect might be important in the case of lower or higher supplementation. ATACa only interfered with some conductance components and this action was predominantly monophasic (decreased or increased conductance). This study indicates differential actions between two closely related molecules on a membranous model.


Entropy provides an unexpected shield in photosynthesis.
Vascular plants combat the excess photon bombarding of high-light conditions with several protective mechanisms. Despite decades of extensive research, new regulatory mech-anisms for photoprotection may remain unknown. Kim et al now report that the monomeric disordered form of photosystem II (PSII), which is present in higher abundance in the native thylakoid membrane in response to high light, possesses an energy-quenching capability superior to that of the multimeric ordered phase, suggesting a new shielding strategy against high-light stress by altering the macro-organization of PSII supercomplexes.


Female genital mutilation (FGM) in Egypt, knowledge and concepts of Egyptian medical students: a cross-sectional study.
There are many factors contributing to the existence of female genital mutilation (FGM). FGM has a long list of recorded complications, which include physical, obstetric, sexual, psychological and death. We assessed the knowledge, attitudes and perceptions of FGM practice among Egyptian medical students. An online questionnaire was distributed. One thousand one hundred and forty-one participants completed questionnaires. 71.2% were aware of FGM complications especially among females. Two-thirds reported illegality. 7.8% were in favour of FGM conduct with a higher male preference. Religious and traditional factors were the principle contributing factors to the practice. Three quarters of female students did not agree that FGM increased the chance of marriage. Almost one-fifth of female students reported having been subject to FGM, with a high level of dissatisfaction. It was shown that Egyptian medical students lacked knowledge about FGM with no structured training, so every effort should be done to end this inhumane practice.Impact StatementWhat is already known on this subject? The conduct of female genital mutilation (FGM) in Egypt is motivated by a variety of factors, including social notions, cultural beliefs and theological misunderstanding. FGM has a long list of recorded complications, which might include physical, obstetric, sexual, psychological and even death.What do the results of this study add? This study provides policy makers and community managers with the evidence needed to advocate for the addition of FGM education to be introduced across the board in medical school curriculums.What are the implications of these findings for clinical practice and/or further research? Possible elimination of the practice and further research on how to eradicate the roots behind it.


Resolving postglacial phylogeography using high-throughput sequencing.
The distinction between model and nonmodel organisms is becoming increasingly blurred. High-throughput, second-generation sequencing approaches are being applied to organisms based on their interesting ecological, physiological, developmental, or evolutionary properties and not on the depth of genetic information available for them. Here, we illustrate this point using a low-cost, efficient technique to determine the fine-scale phylogenetic relationships among recently diverged populations in a species. This application of restriction site-associated DNA tags (RAD tags) reveals previously unresolved genetic structure and direction of evolution in the pitcher plant mosquito, Wyeomyia smithii, from a southern Appalachian Mountain refugium following recession of the Laurentide Ice Sheet at 22,000-19,000 B.P. The RAD tag method can be used to identify detailed patterns of phylogeography in any organism regardless of existing genomic data, and, more broadly, to identify incipient speciation and genome-wide variation in natural populations in general.


[The role and significance of serum SYK and VEGF-C in metastasis of the lymph nodes of pulmonary adenocarcinoma.].
Lung cancer has become one of the most dangerous malignant tumor in the world nowadays, whose pathogenesis is complex involving multi-genes and multi-elements. In this study, we try to explore the values of serum SYK and vascular endothelial growth factor-C (VEGF-C) in lymph node metastasis of lung adenocarcinoma. Serum SYK and VEGF were examined in 160 lung adenocarcinoma patients with various metastasis lesions and 40 healthy volunteers by ELISA. The serum SYK level of the lung adenocarcinoma patients were remarkably lower than those of normal persons (P <0.05); The SYK level decreased while the lymph nodes area increased along with their metastasis (P <0.05), and the serum VEGF-C level were remarkably higher than those of healthy objects (P <0.05). The VEGF-C level increased with the increase of lymph nodes metastasis (P <0.05). For the serum level of SYK and VEGF-C, there was no statistical difference in sex and age (P <0.05). Negative correlation was found between SYK and VEGF-C (r =0.650, P <0.05). The serum level of SYK and VEGF-C are closely related to the lymph nodes metastasis of lung adenocarcinoma. Combination detection of SYK and VEGF-C might be helpful for the predict of lymph nodes metastasis of lung adenocarcinoma.


Detection of an estrogen derivative in two breast cancer cell lines using a single core multimodal probe for imaging (SCoMPI) imaged by a panel of luminescent and vibrational techniques.
3-Methoxy-17α-ethynylestradiol or mestranol is a prodrug for ethynylestradiol and the estrogen component of some oral contraceptive formulations. We demonstrate here that a single core multimodal probe for imaging - SCoMPI - can be efficiently grafted onto mestranol allowing its tracking in two breast cancer cell lines, MDA-MB-231 and MCF-7 fixed cells. Correlative imaging studies based on luminescence (synchrotron UV spectromicroscopy, wide field and confocal fluorescence microscopies) and vibrational (AFMIR, synchrotron FTIR spectromicroscopy, synchrotron-based multiple beam FTIR imaging, confocal Raman microspectroscopy) spectroscopies were consistent with one another and showed a Golgi apparatus distribution of the SCoMPI-mestranol conjugate in both cell lines.


Study of Electrochemical Catalytic Coal Gasification: Gasification Characteristics and Char Structure Evolution.
Electrochemical catalytic coal gasification experiments with Fuxin (FX) coal under a CO2 atmosphere were conducted to evaluate the effects of power and temperature on coal gasification and char structure evolution during electrochemical catalytic gasification (ECG). When the power was 400 W, with temperature increasing from 800 to 1000 °C, the CO content in the gas products increased by 8.16%, the H2 content increased by 8.39%, and the CH4 concentration in the gas products initially increased and then decreased. When the temperature is 900 °C, with power increasing from 0 to 400 W, the CO content in the gas products increased by 58.27%, the H2 content increased by 81.33%, and the CH4 concentration in the gas products increased from 1.31 to 2.37%. The gasification reactivity and the concentration of combustible gas generated during ECG were higher than those during common coal gasification. Thermal electrons play important roles in ECG. These electrons could promote ring opening reactions and aromatic compound cracking and inhibit aromatization reactions while increasing the number of oxygen-containing functional groups in char, consequently enhancing the char gasification reactivity.


Deep Learning-Based Correlation Analysis between the Evaluation Score of English Teaching Quality and the Knowledge Points.
As one of the three main courses from primary school to senior high school, improving the quality of English teaching in and out of class has become the top priority of colleges and universities. English knowledge points are complex, and domestic scholars have studied vocabulary knowledge and grammatical awareness from various perspectives, but there is still a lack of research on the correlation between vocabulary knowledge, grammatical awareness, and cloze test scores of senior high school students. Therefore, this paper carries out empirical research from the depth of English vocabulary, English grammar, reading comprehension, cloze test, and composition, aiming at exploring the relationship between teaching quality and English knowledge points. Classroom teaching quality evaluation is the basic content of education quality evaluation, which not only needs to evaluate the effect of class hours but also needs a student's long-term learning effect. In order to improve the quality of classroom English teaching, enrich the content of classroom English, and ultimately make the quality of students' learning to a higher level, the combination model of the teaching quality evaluation index is established by combining the decision tree with the knowledge point rule association method and the evaluation results are verified by association rule analysis. This paper selects the effective indicators that affect the evaluation of English teaching quality, determines the weight of the indicators by using the analytic hierarchy process, effectively constructs the combination model of the decision tree and rule association method, and establishes the evaluation model of students' English learning ability in the classroom. Taking students as the main object and combining them with the requirements of digital teaching, the evaluation index system is formed, the index weight is determined by using the analytic hierarchy process, and the classroom teaching quality evaluation model based on the decision tree and English knowledge point correlation analysis is constructed to truly reflect the teaching level of teachers, and the correlation analysis is carried out between English teachers' own quality and students' learning effects and knowledge points. By testing the model performance of English vocabulary depth, reading comprehension, grammar, writing, and other knowledge points, we can well evaluate and analyze students' mastery of English learning and the correlation of English knowledge points.


EXpanding Treatment for Existing Neurological Disease (EXTEND): An Open-Label Phase II Clinical Trial of Hydroxyurea Treatment in Sickle Cell Anemia.
Cerebral vasculopathy in sickle cell anemia (SCA) begins in childhood and features intracranial arterial stenosis with high risk of ischemic stroke. Stroke risk can be reduced by transcranial doppler (TCD) screening and chronic transfusion therapy; however, this approach is impractical in many developing countries. Accumulating evidence supports the use of hydroxyurea for the prevention and treatment of cerebrovascular disease in children with SCA. Recently we reported that hydroxyurea significantly reduced the conversion from conditional TCD velocities to abnormal velocities; whether hydroxyurea can be used for children with newly diagnosed severe cerebrovascular disease in place of starting transfusion therapy remains unknown. The primary objective of the EXpanding Treatment for Existing Neurological Disease (EXTEND) trial is to investigate the effect of open label hydroxyurea on the maximum time-averaged mean velocity (TAMV) after 18 months of treatment compared to the pre-treatment value. Secondary objectives include the effects of hydroxyurea on serial TCD velocities, the incidence of neurological and non-neurological events, quality of life (QOL), body composition and metabolism, toxicity and treatment response, changes to brain magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA), genetic and serologic markers of disease severity, and cognitive and pulmonary function. This prospective Phase II trial will enroll children with SCA in Jamaica, between the ages of 2 and 17 years, with either conditional (170-199 cm/sec) or abnormal (≥ 200 cm/sec) TCD velocities. Oral hydroxyurea will be administered daily and escalated to the maximum tolerated dose (MTD). Participants will be seen in the Sickle Cell Unit (SCU) in Kingston, Jamaica monthly until achieving MTD, and then every 3 months. TCD will be performed every 6 months. Currently, 43 participants have been enrolled out of a projected 50. There was one withdrawal due to immigration, with no permanent screen failures. Of the 43 enrolled, 37 participants have initiated study treatment. This trial investigates the effects of hydroxyurea treatment at MTD in children with conditional or abnormal TCD velocities before transfusion therapy and may represent an important advance towards establishing a suitable non-transfusion protocol for stroke prevention in children with SCA. The trial outcomes will have profound significance in developing countries where the disease burden is highest. ClinicalTrials.gov NCT02556099; https://clinicaltrials.gov/ct2/show/NCT02556099 (Archived by WebCite at http://www.webcitation.org/6k1yMAa9G).


Nurses' perspectives of an environment of optimum mental health: a grounded theory study.
The Government's aspiration to make Kenya a middle-income country and achieve the United Nations' sustainable development goal 3, 'good health and well-being', are threatened by poor quality of mental health services. Environment and lack of a conceptual model of nursing to guide care were some of the reasons that were attributed to poor quality of mental health services. The purpose of the study was to discover and describe an appropriate conceptual model of mental health nursing practice. This paper describes the environment which is one of the metaparadigms of a conceptual model grounded on data collected from Kenyan mental health nurses. A grounded theory study was conducted with 33 mental health nurses selected by purposive, open and theoretical sampling procedures. The study sites were level 5 and 6 mental health facilities across the country. Data were collected over a period of 11 months through audio-recorded in-depth interviews and field notes. Analysis was performed using Straussian Grounded Theory steps of open, axial and selective coding processes aided by NVivo version 10. Dimensions and properties of environment metaparadigm grounded on nurses' views were discovered. The study was conducted within the dictates of the institutional and national ethics and research review boards. Environment evolved as an intervening condition and a supra-system for mental health nursing. A homely environment emerged as a space with properties that nurture optimum mental health contrary to a hostile environment that precipitates mental disorders and prolongs recovery. Grounded theory methodology was useful in discovering an environment metaparadigm as the context that influences mental health while nursing practice is the central phenomenon for optimum mental health. Nurses can ensure homely environments from diverse cultures and conduct comparative studies on the recovery of patients in the two environments.


[Application of contrast-enhanced ultrasonography in selecting indication of radiofrequency ablation among hepatocellular carcinoma patients].
To evaluate the clinical value of contrast-enhanced ultrasound (CEUS) in selection of the patients with hepatocellular carcinoma (HCC) indicated to radiofrequency ablation (RFA). 164 patients with HCC, 121 males and 43 females, aged 52.4 (38-72), who asked for RFA were randomly divided into 2 groups: 81 patients undergoing CEUS before RFA (CEUS Group), and 83 patients not undergoing CEUS before RFA (Control Group). There were not significant differences in the TNM staging, liver function Child-Pugh classing, and average tumor size between the 2 groups. Follow-up by conventional ultrasonography, enhanced CT and/or enhanced ultrasonography was conducted for 6-36 months. Nine of the 81 patients in CEUS Group (11.8%) were determined by CEUS as unsuitable for RFA; 5 of the 9 patients had more than 5 lesions, 2 of the 9 patients had lesions > 8 cm in diameter, and the tumor had invaded 2 large vessels or intestine in 2 of the 9 patients. The other 72 patients in CEUS Group, with 101 lesions, underwent RFA after CEUS. CEUS discovered 12 lesions < or = 1.7 cm that had not been discovered by conventional ultrasonography and CT in 8 patients, 3 of which were patients with cirrhosis undergoing follow-up, and 2 of which were HCC patients undergoing follow-up after treatment. Of these 8 patients, 3 had 7 newly discovered satellite lesions around the main lesions. The successful rates of CEUS Group and Control Group were 95.0% and 89.6% respectively (P > 0.05). The distant recurrence rate of Control Group was 22.9%, significantly higher than that of CEUS Group (9.7%, P < 0.05). Helping find minute tumors, CEUS is useful in selecting HCC patients suitable for RFA, thus decreasing the intrahepatic recurrence after RFA.


Multiple cis-elements in the 5'-flanking region of embryonic/larval fast-type of the myosin heavy chain gene of torafugu, MYH(M743-2), function in the transcriptional regulation of its expression.
The myosin heavy chain gene, MYH(M743-2), is highly expressed in fast muscle fibers of torafugu embryos and larvae, suggesting its functional roles for embryonic and larval muscle development. However, the transcriptional regulatory mechanism involved in its expression remained unknown. Here, we analyzed the 2075bp 5'-flanking region of torafugu MYH(M743-2) to examine the spatial and temporal regulation by using transgenic and transient expression techniques in zebrafish embryos. Combining both transient and transgenic analyses, we demonstrated that the 2075bp 5'-flanking sequences was sufficient for its expression in skeletal, craniofacial and pectoral fin muscles. The immunohistochemical observation revealed that the zebrafish larvae from the stable transgenic line consistently expressed enhanced green fluorescent protein (EGFP) in fast muscle fibers. Promoter deletion analyses demonstrated that the minimum 468bp promoter region could direct MYH(M743-2) expression in zebrafish larvae. We discovered that the serum response factor (SRF)-like binding sites are required for promoting MYH(M743-2) expression and myoblast determining factor (MyoD) and myocyte enhancer factor-2 (MEF2) binding sites participate in the transcriptional control of MYH(M743-2) expression in fast skeletal muscles. We further discovered that MyoD binding sites, but not MEF2, participate in the transcriptional regulation of MYH(M743-2) expression in pectoral fin and craniofacial muscles. These results clearly demonstrated that multiple cis-elements in the 5'-flanking region of MYH(M743-2) function in the transcriptional control of its expression.


Cooperation and competition in mismatch repair: very short-patch repair and methyl-directed mismatch repair in Escherichia coli.
In Escherichia coli and related enteric bacteria, repair of base-base mismatches is performed by two overlapping biochemical processes, methyl-directed mismatch repair (MMR) and very short-patch (VSP) repair. While MMR repairs replication errors, VSP repair corrects to C*G mispairs created by 5-methylcytosine deamination to T. The efficiency of the two pathways changes during the bacterial life cycle; MMR is more efficient during exponential growth and VSP repair is more efficient during the stationary phase. VSP repair and MMR share two proteins, MutS and MutL, and although the two repair pathways are not equally dependent on these proteins, their dual use creates a competition within the cells between the repair processes. The structural and biochemical data on the endonuclease that initiates VSP repair, Vsr, suggest that this protein plays a role similar to MutH (also an endonuclease) in MMR. Biochemical and genetic studies of the two repair pathways have helped eliminate certain models for MMR and put restrictions on models that can be developed regarding either repair process. We review here recent information about the biochemistry of both repair processes and describe the balancing act performed by cells to optimize the competing processes during different phases of the bacterial life cycle.


Antisense globin RNA in mouse erythroid tissues: structure, origin, and possible function.
The aim of the experiments described in this paper was to test for the presence of antisense globin RNA in mouse erythroid tissues and, if found, to characterize these molecules. The present study made use of a multistep procedure in which a molecular tag is attached to cellular RNA by ligation with a defined ribooligonucleotide. The act of ligation preserves the termini of RNA molecules, which become the junctions between cellular RNAs and the ligated ribooligonucleotide. It also unambiguously preserves the identity of cellular RNA as a sense or antisense molecule through all subsequent manipulations. Using this approach, we identified and characterized antisense beta-globin RNA in erythroid spleen cells and reticulocytes from anemic mice. We show in this paper that the antisense globin RNA is fully complementary to spliced globin mRNA, indicative of the template/transcript relationship. It terminates at the 5' end with a uridylate stretch, reflecting the presence of poly(A) at the 3' end of the sense globin mRNA. With respect to the structure of their 3' termini, antisense globin RNA can be divided into three categories: full-size molecules corresponding precisely to globin mRNA, truncated molecules lacking predominantly 14 3'-terminal nucleotides, and extended antisense RNA containing 17 additional 3'-terminal nucleotides. The full-size antisense globin RNA contains two 14-nt-long complementary sequences within its 3'-terminal segment corresponding to the 5'-untranslated region of globin mRNA. This, together with the nature of the predominant truncation, suggests a mechanism by which antisense RNA might give rise to new sense-strand globin mRNA.


Bootstrapping Critical Ising Model on Three Dimensional Real Projective Space.
Given conformal data on a flat Euclidean space, we use crosscap conformal bootstrap equations to numerically solve the Lee-Yang model as well as the critical Ising model on a three dimensional real projective space. We check the rapid convergence of our bootstrap program in two dimensions from the exact solutions available. Based on the comparison, we estimate that our systematic error on the numerically solved one-point functions of the critical Ising model on a three dimensional real projective space is less than 1%. Our method opens up a novel way to solve conformal field theories on nontrivial geometries.


Film patterned retarder for stereoscopic three-dimensional display using ink-jet printing method.
We propose a film patterned retarder (FPR) for stereoscopic three-dimensional display with polarization glasses using ink-jet printing method. Conventional FPR process requires coating of photo-alignment and then UV exposure using wire-grid mask, which is very expensive and difficult. The proposed novel fabrication method utilizes a plastic substrate made of polyether sulfone and an alignment layer, poly (4, 4' - (9, 9 -fluorenyl) diphenylene cyclobutanyltetracarboximide) (9FDA/CBDA) in which the former and the latter aligns reactive mesogen along and perpendicular to the rubbing direction, respectively. The ink-jet printing of 9FDA/CBDA line by line allows fabricating the cost effective FPR which can be widely applied for 3D display applications.


Distribution of Mycobacterium avium subspecies paratuberculosis in the lower Florida Keys.
Johne's disease, a fatal and contagious gastrointestinal infection caused by Mycobacterium avium subsp. paratuberculosis (Map), was first diagnosed in an endangered Florida Key deer (Odocoileus virginianus clavium) in 1996 and later in six additional Key deer deaths from 1998 to 2004. We investigated the geographic distribution of Map in the Lower Florida Keys from February 2005 through May 2006 via collection of blood and fecal pellets from 51 live-captured deer, collection of 550 fecal samples from the ground, and by necropsies of 90 carcasses. Tissue and fecal samples also were submitted from 30 raccoons (Procyon lotor), three feral cats (Felis catus), an opossum (Didelphis virginiana), and a Lower Keys marsh rabbit (Sylvilagus palustris hefneri). Mycobacterium avium subsp. paratuberculosis was identified in 23 Key deer fecal samples collected from the ground, tissue samples from two clinically ill Key deer, and from the mesenteric lymph node of a raccoon. The results of this study indicate that Map persists in the Key deer population and environment at a low prevalence, but its distribution currently is limited to a relatively small geographic area within the range of Key deer.


Interaction between HDV and HBV infection in HBsAg-chronic carriers.
We studied the interaction between HBV and HDV infection in 149 consecutive subjects with HBsAg positive chronic hepatitis and in 22 chronic HBsAg healthy carriers. Liver HBcAg was detected in 52 (30.4%) of the 171 subjects. Of these 52, 35 were HBV-DNA and HBeAg positive, 11 HBV-DNA positive only; two HBeAg positive only and four were negative for both HBeAg and HBV-DNA. None of the 119 HBcAg-negative subjects had detectable HBV-DNA in serum. HD-Ag in hepatocytes was detected in 31 of the 171 subjects (18%); it was detectable in none of the 22 HBsAg healthy carriers, in four of the 56 patients with chronic persistent hepatitis (7.2%), in six of the 24 patients with chronic lobular hepatitis (25%), in 16 of the 40 patients with chronic active hepatitis (40%) and in five of the 29 with cirrhosis (17%). A presence of anti-HD in serum in the absence of liver HD-Ag was found in 54 of the 171 subjects (32%). This condition was observed not only in patients with a progressive disease (37.7% of chronic active hepatitis or cirrhosis and 33% of chronic lobular hepatitis), but also in healthy carriers (36%) and in chronic persistent hepatitis patients (21.4%). Liver HBcAg was detected in 6.4% of the 31 HD-Ag-positive patients, in 12.9% of the 54 HD-Ag-negative/anti-HD positive, but in 50% of the 86 with no marker of HDV infection. HDV appears to inhibit HBV genome and such inhibition may persist even when anti-HD is the only HDV marker detectable.


In vitro modulation of oral bacterial adhesion to saliva-coated hydroxyapatite beads by milk casein derivatives.
Bovine caseinate, derivatives of its glycosylated moiety [caseinoglycomacropeptide (CGP)], and caseinophosphopeptides were evaluated as inhibitors of adhesion of oral bacteria to saliva-coated hydroxyapatite beads (S-HA). All milk casein-derived components behaved as potent inhibitors of Streptococcus sanguis OMZ 9 and Streptococcus sobrinus OMZ 176 adhesion to S-HA, whereas neither bovine serum albumin nor polyethyleneglycol were able to interfere with the adhesion of these strains. By contrast, none of the molecular species tested was able to inhibit the attachment of Actinomyces viscosus Ny 1 to S-HA. On the other hand, casein derivatives were shown to displace human serum albumin from S-HA beads. They were also able to bind to the bacterial cell surface of all strains examined. Collectively, these findings suggest that interactions between acidic casein-derived milk components and the biological surfaces involved in bacterial adhesion to S-HA result in an inhibitory effect that is selective for the oral streptococci examined.


A randomized double blind placebo controlled clinical trial of N-Acetylcysteine added to risperidone for treating autistic disorders.
This study examined the efficacy and safety of N-acetylcysteine (NAC) augmentation for treating irritability in children and adolescents with autism spectrum disorders (ASD). Forty children and adolescents met diagnostic criteria for ASD according to DSM-IV. They were randomly allocated into one of the two groups of NAC (1200 mg/day)+risperidone or placebo+risperidone. NAC and placebo were administered in the form of effervescent and in two divided doses for 8 weeks. Irritability subscale score of Aberrant Behavior Checklist (ABC) was considered as the main outcome measure. Adverse effects were also checked. The mean score of irritability in the NAC+risperidone and placebo+risperidone groups at baseline was 13.2(5.3) and 16.7(7.8), respectively. The scores after 8 weeks were 9.7(4.1) and 15.1(7.8), respectively. Repeated measures of ANOVA showed that there was a significant difference between the two groups after 8 weeks. The most common adverse effects in the NAC+risperidone group were constipation (16.1%), increased appetite (16.1%), fatigue (12.9%), nervousness (12.9%), and daytime drowsiness (12.9%). There was no fatal adverse effect. Risperidone plus NAC more than risperidone plus placebo decreased irritability in children and adolescents with ASD. Meanwhile, it did not change the core symptoms of autism. Adverse effects were not common and NAC was generally tolerated well. This trial was registered at http://www.irct.ir. The registration number of this trial was IRCT201106103930N6.


Gastroschisis and Hirschsprung's disease: a rare combination.
The authors report on a neonate with gastroschisis repaired at birth who later had abdominal distension, emesis, feeding intolerance, and an abnormal stooling pattern. Total colon and partial small bowel aganglionosis (TCAS), or Hirschsprung's disease, was diagnosed subsequently. This is the first report of this combination of gastrointestinal anomalies. J Pediatr Surg 36:638-640.


Structural and functional characterization of a highly stable endo-β-1,4-xylanase from Fusarium oxysporum and its development as an efficient immobilized biocatalyst.
Replacing fossil fuel with renewable sources such as lignocellulosic biomass is currently a promising alternative for obtaining biofuel and for fighting against the consequences of climate change. However, the recalcitrant structure of lignocellulosic biomass residues constitutes a major limitation for its widespread use in industry. The efficient hydrolysis of lignocellulosic materials requires the complementary action of multiple enzymes including xylanases and β-xylosidases, which are responsible for cleaving exo- and endoxylan linkages, that release oligocarbohydrates that can be further processed by other enzymes. We have identified the endo-β-1,4-xylanase Xyl2 from Fusarium oxysporum as a promising glycoside hydrolase family 11 enzyme for the industrial degradation of xylan. To characterize Xyl2, we have cloned the synthetic optimized gene and expressed and purified recombinant Xyl2 to homogeneity, finally obtaining 10 mg pure Xyl2 per liter of culture. The crystal structure of Xyl2 at 1.56 Å resolution and the structure of a methyl-xylopyranoside Xyl2 complex at 2.84 Å resolution cast a highly detailed view of the active site of the enzyme, revealing the molecular basis for the high catalytic efficiency of Xyl2. The kinetic analysis of Xyl2 demonstrates high xylanase activity and non-negligible β-xylosidase activity under a variety of experimental conditions including alkaline pH and elevated temperature. Immobilizing Xyl2 on a variety of solid supports enhances the enzymatic properties that render Xyl2 a promising industrial biocatalyst, which, together with the detailed structural data, may establish Xyl2 as a platform for future developments of industrially relevant xylanases. F. oxysporum Xyl2 is a GH11 xylanase which is highly active in free form and immobilized onto a variety of solid supports in a wide pH range. Furthermore, immobilization of Xyl2 on certain supports significantly increases its thermal stability. A mechanistic rationale for Xyl2's remarkable catalytic efficiency at alkaline pH is proposed on the basis of two crystallographic structures. Together, these properties render Xyl2 an attractive biocatalyst for the sustainable industrial degradation of xylan.


Polyphosphate Hydrolysis within Acidic Vacuoles in Response to Amine-Induced Alkaline Stress in the Halotolerant Alga Dunaliella salina.
The location and mobilization of polyphosphates in response to an amine-induced alkaline stress were studied in the halotolerant alga Dunaliella salina. The following observations suggest that polyphosphates accumulate in acidic vacuoles: (a) Accumulation of large amounts of polyphosphates is manifested as intravacuolar dense osmiophilic bodies in electron micrographs. (b) Uptake of amines into the vacuoles induces massive hydrolysis of polyphosphates, demonstrated by in vivo(31)P-nuclear magnetic resonance, and by analysis of hydrolytic products on thin layer chromatograms. The analysis indicates that: (a) Polyphosphate hydrolysis is kinetically correlated with amine accumulation and with the recovery of cytoplasmic pH. (b) The major hydrolytic product is tripolyphosphate. (c) The peak position of the tripolyphosphate terminal phosphate in nuclear magnetic resonance spectra is progressively shifted as the cells recover, indicating that the pH inside the vacuoles increases while the pH in the cytoplasm decreases. (d) In lysed cell preparations, in which vacuoles become exposed to the external pH, mild alkalinization in the absence of amines induces polyphosphate hydrolysis to tripolyphosphates. It is suggested that amine accumulation within vacuoles activates a specific phosphatase, which hydrolyzes long-chain polyphosphates to tripolyphosphates. The hydrolysis increases the capacity of the vacuoles to sequester amines from the cytoplasm probably by releasing protons required to buffer the amine, and leads to recovery of cytoplasmic pH. Thus, polyphosphate hydrolysis provides a high-capacity buffering system that sustains amine compartmentation into vacuoles and protects cytoplasmic pH.


[Infection control in the intensive care unit].
The basis of infection control is systematic monitoring of infections in the patient care units, whereby a clear-cut basis for focusing of the control is obtained. An entity of several distinct evidence-based methods, i.e. treatment bundle, is increasingly utilized. Though difficult in practice, the goal is zero tolerance, whereby all infections during the treatment should be prevented. Central principles of infection control include use of antimicrobial hand rinse in all patient care, appropriate use of protective devices and contact isolation as required by the detected microorganisms.


Phakic intraocular lens to correct high myopic amblyopia in children.
In a clinical investigation, we evaluated anatomical and functional outcomes of posterior phakic chamber lens (ICL) implantation for correction of high myopia with amblyopia in children. Twelve eyes of 11 children, age 3 to 16 years, with high myopic amblyopia were operated with implantation of a Staar Surgical ICL. In these patients, conventional therapy with spectacles or contact lenses was unsuccessful. Mean preoperative spherical equivalent refraction was -12.70 D (range -8.00 to -18.00 D) and best spectacle-corrected visual acuity ranged from count fingers to 20/63. Mean follow-up was 20.5 months (range 3 to 48 mo) Preoperative and postoperative anatomical and functional outcomes were compared. We noted good tolerance of ICLs without inflammatory reactions or secondary capsular opacity, stable intraocular pressure, and good ICL position in all eyes. Predictability was +0.71 D (range -0.75 to +2.00 D). Mean postoperative best spectacle-corrected visual acuity was 20/63. Recovery of binocular vision was achieved in six patients and orthotropic position in seven patients. Quality of life was improved in all patients. The Staar Surgical phakic ICL appeared to be an effective method to treat high myopia in children with amblyopia. Good results with high satisfaction were noted.


[Experimental study of the suture of the intraocular lens sutured to the ciliary sulcus. Needle passage and distance from the limbus].
To discover which technique is best to scleral suture fixation of the lens in the ciliary sulcus, based on two needle direction options: passing the needle from the outer part of the eyeball inwards or from the inner part outwards and to know at what distance the needle must be passed from the sclerocorneal limbus. We used 40 cadaver eyes conserved in 10% formaldehyde. The needle was passed from the interior to the exterior of the eye, observing whether if it had crossed through the ciliary sulcus and the distance of the needle from the limbus. Stitches were also made, going from the exterior to the interior of the eyeball at a known distance from the limbus, either parallel to the iris or perpendicular to the sclera. When the needle was passed from the interior toward the exterior of the eye, it passed through the ciliary sulcus, exiting the eye at 1.50+/-0.16 mm from the limbus, in 80% of the cases. When the needle was passed from the exterior toward the interior of the globe perpendicularly to the sclera only 32.5% passed through the sulcus and when it was parallel to the iris only, only 40%. There is greater statistical probability that the needle will pass through the sulcus from the interior of the eyeball, exiting the limbus at approximately 1.5 mm. Since it is necessary to pass the suture through the sulcus in order to be able to place the lens haptics in the ciliary sulcus when suturing a posterior chamber lens to the sclera, it is best to pass the needle from the interior to the exterior of the eyeball and for the needle to exit at approximately 1.5 mm from the limbus.


[Herpes simplex virus pneumonia].
Herpes simplex virus 1 (HSV-1) reactivation is frequent in intensive care unit patients (20% to 50%, depending on case-mix): it begins in the throat after 3 to 5 days of hospital stay, then progress towards the respiratory tract to reach the lung after 7 to 10 days. In few cases, a HSV-1 bronchopneumonitis (defined by clinical symptom, HSV-1 in the lower respiratory tract and either specific cytopathic effect on cells collected during bronchoalveolar lavage or a HSV-1 load above 105 copies/million cells), can occur. HSV-1 reactivation is associated with mortality, but whether it is only a bystander or it has its own morbidity/mortality is unknown. Two randomized trials have shown lack of efficacy of acyclovir prophylactic or preemptive treatment, but observational studies suggest that acyclovir may be beneficial as a curative treatment. To date, we can only recommend acyclovir in patients with HSV-1 bronchopneumonitis or with HSV-1 load above 105 copies/million cells in the bronchoalveolar lavage.


Haemogram-Derived Indices for Screening and Prognostication in Critically Ill Septic Shock Patients: A Case-Control Study.
This study aimed (1) to assess the diagnostic accuracy of neutrophil-to-lymphocyte (NLR), platelet-to-lymphocyte (PLR), monocyte-to-lymphocyte (MLR) and platelet count-to-mean platelet volume (PLT/MPV) ratios in predicting septic shock in patients on admission to the intensive care unit (ICU) and (2) to compare it with the role of C-reactive protein (CRP), procalcitonin (PCT) and lactate level. We also sought (3) to verify whether the indices could be useful in ICU mortality prediction and (4) to compare them with Acute Physiology and Chronic Health Evaluation II (APACHE II), Simplified Acute Physiology Score II (SAPS II) and Sequential Organ Failure Assessment (SOFA) scores. This retrospective study covered 138 patients, including 61 subjects with multi-organ failure due to septic shock (study group) and 77 sex- and age-matched controls. Septic patients had significantly higher NLR (p < 0.01) and NLR predicted septic shock occurrence (area under the ROC curve, AUROC = 0.66; 95% CI 0.58-0.74). PLR, MLR and PLT/MPV were impractical in sepsis prediction. Combination of CRP with NLR improved septic shock prediction (AUROC = 0.88; 95% CI 0.81-0.93). All indices failed to predict ICU mortality. APACHE II and SAPS II predicted mortality with AUROC = 0.68; 95% CI 0.54-0.78 and AUROC = 0.7; 95% CI 0.57-0.81, respectively. High NLR may be useful to identify patients with multi-organ failure due to septic shock but should be interpreted along with CRP or PCT. The investigated indices are not related with mortality in this specific clinical setting.


Core promoter elements of eukaryotic genes have a highly distinctive mechanical property.
In spite of the abundant data on DNA sequence, the mechanical aspects of promoter DNA remain poorly understood. We classified 1871 human and 196 mouse RNA polymerase II promoters and investigated average flexibility profiles of the human promoters containing either a TATA box or an initiator (Inr) sequence only. Here, we show that TATA boxes and Inr sequences have a common anomalous mechanical property: they are comprised of distinctively flexible and rigid sequences, compared with the other parts of the promoter region. The +2 position in the Inr consensus sequence does not favor adenine to keep the high flexibility and thus this position is more accurately represented as 'T, G, C>>A'. Additionally, it was also found that DNA region upstream of TATA box or Inr sequence is more rigid than region downstream of each element. These properties may function as a marker for recognition by TATA-binding protein and Inr-binding protein.


Analysis of Predictive Biomarkers in Patients With Lung Adenocarcinoma From Southern Brazil Reveals a Distinct Profile From Other Regions of the Country.
Adenocarcinoma is the most common histologic subtype of non-small-cell lung cancer, representing 40% of all diagnoses. Several biomarkers are currently used to determine patient eligibility for targeted treatments, including analysis of molecular alterations in EGFR and ALK, as well as programmed death-ligand 1 (PD-L1) protein expression. Epidemiologic data reporting the frequency of these biomarkers in Brazilian patients with lung adenocarcinoma (LUAD) are limited, and existing studies predominantly included patients from the southeast region of the country. The goal of this study was to investigate the frequency of somatic mutations in the EGFR, KRAS, NRAS, and BRAF genes, ALK, and PD-L1 expression in a series of Brazilian patients diagnosed with LUAD predominantly recruited from centers in southern Brazil. Molecular analysis of the EGFR, KRAS, NRAS, and BRAF genes was performed by next-generation sequencing using DNA extracted from tumor tissue. Immunohistochemistry was used to detect ALK and PD-L1 expression. Analysis of 619 tumors identified KRAS mutations in 189 (30.2%), EGFR mutations in 120 (19.16%), and BRAF mutations in 19 (3%). Immunohistochemistry demonstrated ALK and PD-L1 expression in 4% and 35.1% of patients, respectively. To our knowledge, this is the first study investigating the molecular epidemiology of patients with LUAD from southern Brazil and the largest assessing the frequency of multiple predictive biomarkers for this tumor in the country. The study also reveals a distinct mutation profile compared with data originating from other regions of Brazil.


Is niacin ineffective? Or did AIM-HIGH miss its target?
The AIM-HIGH trial (Atherothrombosis Intervention in Metabolic Syndrome With Low HDL/High Triglycerides: Impact on Global Health Outcomes) found, in an interim analysis, no cardiovascular benefit from taking extended-release niacin (Niaspan). In fact, there was a trend toward a greater risk of ischemic stroke, which did not reach statistical significance. But questions remain about this complex trial, which included intensive statin therapy in the active-treatment group and the control group.


Comparison of rat pulmonary and hepatic cytosolic alcohol dehydrogenase activities.
The liver is the major organ responsible for ethanol oxidation, and alcohol dehydrogenase (ADH) is the main enzyme involved. There is limited evidence suggesting the involvement of the lung in ethanol metabolism. To determine the degree to which pulmonary ADH plays a role in ethanol metabolism, ADH activity was measured spectrophotometrically using hepatic and pulmonary cytosolic fractions prepared by differential centrifugation and Sephadex G-50 column chromatography. Apparent Km values for hepatic and pulmonary ADHs were determined. Inhibition constants were calculated using 4-methylpyrazole. The ADHs were characterized by examining the influence of pH on enzyme activity. Pulmonary ADH activity was much lower at near neutral pH than at pH 9.0 or 10, whereas hepatic ADH activity was also pH dependent but was significantly higher. Pulmonary ADH is less sensitive to inhibition by 4-methylpyrazole than is hepatic ADH, as evidenced by a 1000-fold higher Ki. Pulmonary ADH would be expected to make only a minor contribution to ethanol metabolism in vivo.


Exploring soft matter: liposomal vesicles and their tethers.
The malleability of soft matter such as vesicular systems, allows their mechanical manipulation into various interesting structures. Their inherent elastic membrane properties can be utilized in fabricating micro-scaled transport machineries. In this paper, we have described several features of research carried out in our laboratory. This includes the self-healing ability of multilamellar liposome on removal of their inner core. The formation of tethers from their outer bilayers into complex networks has been demonstrated. Photo enhancement of images has revealed the apparent existence of narrow channels inside such tethers. The tethers channels were subsequently used as transport conduits to carry solid polystyrene microspheres as well as large liposomes larger than the tether diameter under controllable speeds of up to 2 microm s(- 1). The possibility of producing multi-compartment liposomal systems connected to each other directly by fusion or indirectly via tether networks has provided us with a system capable of transporting components from one vesicle entity to another.


Weight-based dosing in medication use: what should we know?
Weight-based dosing strategy is still challenging due to poor awareness and adherence. It is necessary to let clinicians know of the latest developments in this respect and the correct circumstances in which weight-based dosing is of clinical relevance. A literature search was conducted using PubMed. Clinical indications, physiological factors, and types of medication may determine the applicability of weight-based dosing. In some cases, the weight effect may be minimal or the proper dosage can only be determined when weight is combined with other factors. Medications within similar therapeutic or structural class (eg, anticoagulants, antitumor necrosis factor medications, P2Y12-receptor antagonists, and anti-epidermal growth factor receptor antibodies) may exhibit differences in requirements on weight-based dosing. In some cases, weight-based dosing is superior to currently recommended fixed-dose regimen in adult patients (eg, hydrocortisone, vancomycin, linezolid, and aprotinin). On the contrary, fixed dosing is noninferior to or even better than currently recommended weight-based regimen in adult patients in some cases (eg, cyclosporine microemulsion, recombinant activated Factor VII, and epoetin α). Ideal body-weight-based dosing may be superior to the currently recommended total body-weight-based regimen (eg, atracurium and rocuronium). For dosing in pediatrics, whether weight-based dosing is better than body surface-area-based dosing is dependent on the particular medication (eg, methotrexate, prednisone, prednisolone, zidovudine, didanosine, growth hormone, and 13-cis-retinoic acid). Age-based dosing strategy is better than weight-based dosing in some cases (eg, intravenous busulfan and dalteparin). Dosing guided by pharmacogenetic testing did not show pharmacoeconomic advantage over weight-adjusted dosing of 6-mercaptopurine. The common viewpoint (ie, pediatric patients should be dosed on the basis of body weight) is not always correct. Effective weight-based dosing interventions include standardization of weight estimation, documentation and dosing determination, dosing chart, dosing protocol, order set, pharmacist participation, technological information, and educational measures. Although dosing methods are specified in prescribing information for each drug and there are no principal pros and cons to be elaborated, this review of weight-based dosing strategy will enrich the knowledge of medication administration from the perspectives of safety, efficacy, and pharmacoeconomics, and will also provide research opportunities in clinical practice. Clinicians should be familiar with dosage and administration of the medication to be prescribed as well as the latest developments.


Development of the fingerprint for the quality of Radix Linderae through ultra-pressure liquid chromatography-photodiode array detection/electrospray ionization mass spectrometry.
An ultra-pressure LC (UPLC) method has been developed and validated for the quality evaluation of a traditional Chinese medicine (Radix Linderae) by chemical fingerprint analysis with chromatograms collected at two wavelengths (260 and 320 nm). Eleven characteristic peaks in the fingerprints were identified by comparing their retention times, UV spectra and ESI-MS/MS data with those of the reference substances or the data in the literatures. Both correlation coefficient of similarities in chromatograms and relative peak areas of common peaks were calculated for quality expression of the Radix Linderae samples collected from different areas in China. The results showed high variation of relative peak area and correlation coefficients among the samples collected from various habitats, which indicated that the quality consistency of Radix Linderae is still a problem worthy of serious concern.


Patients with a phenotype consistent with facioscapulohumeral muscular dystrophy display genetic and epigenetic heterogeneity.
To identify the genetic and epigenetic defects in patients presenting with a facioscapulohumeral (FSHD) clinical phenotype without D4Z4 contractions on chromosome 4q35 tested by linear gel electrophoresis and Southern blot analysis. The authors studied 16 patients displaying an FSHD-like phenotype, with normal cardiovascular and respiratory function, a myopathic pattern on electromyography, and a muscle biopsy being normal or displaying only mild and aspecific dystrophic changes. They sequenced the genes calpain 3 (CAPN3), valosin containing protein (VCP) and four-and-a-half LIM domains protein 1 (FHL1), and they analysed the D4Z4 repeat arrays by extensive genotyping and DNA methylation analysis. The authors identified one patient carrying a complex rearrangement in the FSHD locus that masked the D4Z4 contraction associated with FSHD1 in standard genetic testing, one patient with somatic mosaicism for the D4Z4 4q35 contraction, six patients that were diagnosed as having FSHD2, four patients with CAPN3 mutations and two patients with a VCP mutation, No mutations were detected in FHL1, and in two patients, the authors could not identify the genetic defect. In patients presenting with an FSHD-like clinical phenotype with a negative molecular testing for FSHD, consider (1) detailed genetic testing including D4Z4 contraction of permissive hybrid D4Z4 repeat arrays, p13E-11 probe deletions, and D4Z4 hypomethylation in the absence of repeat contraction as observed in FSHD2; (2) mutations in CAPN3 even in the absence of protein deficiency on western blot analysis; and (3) VCP mutations even in the absence of cognitive impairment, Paget disease and typical inclusion in muscle biopsy.


Structure-activity relationships of the inhibitory effects of flavonoids on P-glycoprotein-mediated transport in KB-C2 cells.
We studied the effects of flavonoids, naringenin (flavanone), baicalein (flavone), kaempferol, quercetin, myricetin, morin, and fisetin (flavonols) as well as two glycosides of quercetin on P-glycoprotein (P-gp) function in multidrug-resistant P-gp overexpressing KB-C2 cells. Flavonoids such as kaempferol and quercetin increased the accumulation of rhodamine-123 dependent on their chemical structure. Analysis by flow cytometry indicated that the increase in substrate accumulation was due to the inhibition of substrate efflux. Naringenin, which lacks the 2,3-double bond in the C ring, had no effect, although it was more hydrophobic than myricetin, fisetin and morin. Therefore, the planar structure of the flavonoids seemed to be important for their interaction with P-gp. The effects of other flavonoids on the accumulation of daunorubicin were in the order of kaempferol>quercetin, baicalein>myricetin>fisetin, morin. Quercetin-3-O-glucoside and rutin had no effect. The order of the effects corresponded with that of the partition coefficients. Difference in the number and position of hydroxyl groups in flavonoid molecules by themselves seemed to have little effect. These results suggested that hydrophobicity as well as planar structure is important for the inhibitory effects of flavonoids on P-gp-mediated transport.


Fungal Keratitis Caused by Drechslera spp. Treated with Voriconazole: A Case Report.
Objective. To present a case of Drechslera spp. keratitis treated with topical Voriconazole. Method. A case report. Results. A 52-year-old diabetic male presented with a one-week history of foreign body sensation of the left eye, self-medicated with Neomycin, Polymyxin B, and Dexamethasone eye drops, and was diagnosed to have bacterial conjunctivitis, which was treated with Levofloxacin drops. The patient developed a corneal opacity after 2 days and was initially seen with a visual acuity of counting fingers on the left eye, with a 3 mm central corneal ulcer with feathery borders. No hypopyon was noted. The right eye had a visual acuity of 20/20 and had unremarkable findings. Corneal scraping of the ulcer showed no organisms on Gram and Giemsa stain. Cultures were positive for Drechslera spp. and patient was started on Natamycin drops every 15 minutes, Atropine drops 3× a day, and Levofloxacin was continued every 4 hours. The ulcer increased to 4 mm, the infiltrates became deeper involving the midstroma, and there was appearance of a 2 mm hypopyon. Natamycin was shifted to Voriconazole eye drops every 15 minutes. There was note of a decrease in the size of the ulcer and clearing of the infiltrates with the new treatment regimen. Final visual acuity after 29 days of treatment was 20/40 with note of a slight corneal haze in the area of the previous ulcer. Conclusion. Voriconazole may be safe and effective in the treatment of Drechslera keratitis. There was no perforation and there was immediate decrease in the size of the ulcer. This is the first known case of Drechslera keratitis treated with Voriconazole eye drops in the Philippines.


Effect of Mono- and Divalent Metal Ions on Current-Voltage Features of a λ-DNA Solution Electrically Driven in a Microfluidic Capillary.
The interactions of DNA molecules and metal ions lead to changes in their configuration and conformation, which in turn influence the current characteristics of the solution as DNA molecules are translocated through a micro/nanofluidic channel and ultimately cause serious impacts on the practical applications of DNA/gene chips for precisely manipulating and studying the molecular properties of single DNA molecules. In this study, the current characteristics of λ-DNA solutions without or with metal ions (i.e., K+, Na+, Mg2+, and Ca2+) were experimentally investigated when they were transported through a 5 μm microcapillary under an external electric field with asymmetric electrodes. Experimental data indicated some meaningful results. First, the current-voltage relations of the metal ion solutions were all linear, while those of λ-DNA solutions without or with metal ions were all nonlinear and followed power functions, of which the indices were related to the type, valence, and mobility of ions. Furthermore, as the concentrations of metal ions increased, the power indices of the λ-DNA solutions with monovalent metal ions increased, while those of the λ-DNA solutions with divalent ions decreased. Finally, the main reasons for the current characteristics were theoretically attributed to two possible mechanisms: the polarizations on the asymmetric electrodes and the interactions between λ-DNA and metal ions. These findings are helpful for the design of new biomedical micro/nanofluidic sensors and labs on a chip for accurately manipulating single DNA molecules.


Integration of Wet-Lab Measures, Milk Infrared Spectra, and Genomics to Improve Difficult-to-Measure Traits in Dairy Cattle Populations.
The objective of this study was to evaluate the contribution of Fourier-transformed infrared spectroscopy (FTIR) data for dairy cattle breeding through two different approaches: (i) estimating the genetic parameters for 30 measured milk traits and their FTIR predictions and investigating the additive genetic correlation between them and (ii) evaluating the effectiveness of FTIR-derived phenotyping to replicate a candidate bull's progeny testing or breeding value prediction at birth. Records were available from 1,123 cows phenotyped using gold standard laboratory methodologies (LAB data). This included phenotypes related to fine milk composition and milk technological characteristics, milk acidity, and milk protein fractions. The dataset used to generate FTIR predictions comprised 729,202 test-day records from 51,059 Brown Swiss cows (FIELD data). A first approach consisted of estimating genetic parameters for phenotypes available from LAB and FIELD datasets. To do so, a set of bivariate animal models were run, and genetic correlations between LAB and FIELD phenotypes were estimated using FIELD information obtained at the population level. Heritability estimates were generally higher for FIELD predictions than for the corresponding LAB measures. The additive genetic correlations (r a ) between LAB and FIELD phenotypes had different magnitudes across traits but were generally strong. Overall, these results demonstrated the potential of using FIELD information as indicator traits for the indirect genetic improvement of LAB measures. In the second approach, we included genotype information for 1,011 cows from the LAB dataset, 1,493 cows from the FIELD dataset, 181 sires with daughters in both LAB and FIELD datasets, and 540 sires with daughters in the FIELD dataset only. Predictions were obtained using the single-step GBLUP method. A four fold cross-validation was used to assess the predictive ability of the different models, assessed as the ability to predict masked LAB records from daughters of progeny testing bulls. The correlation between observed and predicted LAB measures in validation was averaged over the four training-validation sets. Different sets of phenotypic information were used sequentially in cross-validation schemes: (i) LAB cows from the training set; (ii) FIELD cows from the training set; and (iii) FIELD cows from the validation set. Models that included FIELD records showed an improvement for the majority of traits. This study suggests that breeding programs for difficult-to-measure traits could be implemented using FTIR information. While these programs should use progeny testing, acceptable values of accuracy can be achieved also for bulls without phenotyped progeny. Robust calibration equations are, deemed as essential.


[Cost-effectiveness of photodynamic therapy in age-related macular degeneration].
The aim of this study was to estimate the public health service cost of visual acuity improvement or maintenance with photodynamic therapy in patients with age-related macular degeneration (ARMD). This illness is the most frequent cause of blindness in elderly patients in western countries. A cost-effectiveness analysis was carried out to compare photodynamic therapy versus no treatment. The analysis point of view was that of the health service. The improvement or maintenance of visual acuity and contrast sensitivity were considered efficacy results. Direct costs were estimated by means of cost accountancy. Quality adjusted costs per visual acuity life year gained (QACVAG) were calculated through utility values from other studies. The cost per year of maintenance of visual acuity in a two-year period was 36,530 euro for women and 34,804 euro for men. If this cost was estimated for life expectancy in Asturias, it would be reduced to 4,298 euro for women and 5,354 euro for men. If costs of the QACVAG, in a two-year period, were considered, photodynamic therapy would cost 66,931 euro for women and 70,249 euro for men. This cost-effectiveness analysis allows decisions to be made about public financing. Some research in our country suggests that public health financing should be provided for interventions whose cost-effectiveness is less than 30,000 euro of CVAQA. The treatment evaluated here far exceeds this value. It is recommended that the use of more restrictive patient selection, incorporating diagnostic criteria and patient autonomy indicators, could improve the results of this intervention.


[Isolation and various properties of nucleoside monophosphate kinases from Escherichia coli].
A technique is proposed for isolation of nucleosidemonophosphate kinases--AMP-kinase (EC 220.127.116.11), GMP-kinase (EC 188.8.131.52), CMP-kinase (EC 22.214.171.124), UMP-kinase (EC 220.127.116.11) and TMP-kinase (EC 126.96.36.199)--from E. coli MRE-600. It involves cell destroying, precipitation of nucleic acids with polyethyleneimine, fractionation with ammonium sulphate followed by chromatography on different carriers (DEAE-Toyopearl-650 M, Matrex gel Blue A, Matrex gel Red A). The technique enables all the five enzymes to be obtained separately and without contaminations with nucleotide dephosphorylating enzymes. For all the enzymes the pH optimum was found to range from 6.5 to 8.0, and Mg2+ ions were found to be the best activator for all the enzymes studied. The substrate specificity was investigated with respect to acceptors and donors of the phosphate groups. The enzymes showed strict specificity to the heterocyclic base of the acceptor phosphate group. AMP-, GMP- and CMP-kinases phosphorylated the corresponding deoxynucleoside monophosphates less effectively than ribonucleoside monophosphates. ATP was found to be the most effective phosphate donor for all the enzymes under study.


An evidence-based review of the scope and potential ethical concerns of teleorthodontics.
The aim was to evaluate the scope and potential ethical concerns related to the use of teledentistry in clinical orthodontics. Indexed databases were searched up to and including October 2020. The eligibility criteria were as follows: (a) original clinical studies, and (b) case reports/series. Historic reviews, commentaries, experimental studies, and letters to the editor, were excluded. The pattern of the present review was customized to summarize the relevant information. A total of 4 clinical studies (out of 1016) were included in the present evidence-based review. Three studies reported that teledentistry is useful in clinical orthodontics. In 1 study, a clear conclusion could not be derived regarding the benefits of teledentistry in clinical orthodontics. Two out of 4 studies did not obtain prior approval from an Institutional Review Board or Ethical Committee. Three studies did not report any measures that were undertaken to safeguard the electronic transfer of patient-related health information. Teleorthodontics facilitates treatment planning/monitoring by sharing orthodontics-based patient records among oral healthcare providers; however, the importance of direct patient supervision and routine follow-ups during orthodontic therapy cannot be overlooked. Further studies are needed to establish ethical guidelines and a standard of care in this emerging field.


Isolated mouse islets respond to glucose with an initial peak of glucagon release followed by pulses of insulin and somatostatin in antisynchrony with glucagon.
Recent studies of isolated human islets have shown that glucose induces hormone release with repetitive pulses of insulin and somatostatin in antisynchrony with those of glucagon. Since the mouse is the most important animal model we studied the temporal relation between hormones released from mouse islets. Batches of 5-10 islets were perifused and the hormones measured with radioimmunoassay in 30s fractions. At 3mM glucose, hormone secretion was stable with no detectable pulses of glucagon, insulin or somatostatin. Increase of glucose to 20mM resulted in an early secretory phase with a glucagon peak followed by peaks of insulin and somatostatin. Subsequent hormone secretion was pulsatile with a periodicity of 5min. Cross-correlation analyses showed that the glucagon pulses were antisynchronous to those of insulin and somatostatin. In contrast to the marked stimulation of insulin and somatostatin secretion, the pulsatility resulted in inhibition of overall glucagon release. The cytoarchitecture of mouse islets differs from that of human islets, which may affect the interactions between the hormone-producing cells. Although indicating that paracrine regulation is important for the characteristic patterns of pulsatile hormone secretion, the mouse data mimic those of human islets with more than 20-fold variations of the insulin/glucagon ratio. The data indicate that the mouse serves as an appropriate animal model for studying the temporal relation between the islet hormones controlling glucose production in the liver.


Enhancement of biogas production from swine manure by a lignocellulolytic microbial consortium.
Anaerobic digestion of lignocellulosic wastes is limited by inefficient hydrolysis of recalcitrant substrates, leading to low biogas yield. In this study, the potential of a lignocellulolytic microbial consortium (LMC) for enhancing biogas production from fibre-rich swine manure (SM) was assessed. Biochemical methane potential assay showed that inoculation of structurally stable LMC to anaerobic digestion led to increase biogas production under mesophilic and thermophilic conditions. The greatest enhancement was observed at 37°C with a LMC/SM ratio of 1.5:1 mg VSS/g VS leading to biogas and methane yields of 355 and 180 ml/g VS(added) respectively, equivalent to 40% and 55% increases compared with the control. The LMC was shown to increase the efficiency of total solid, chemical oxygen demand removal and degradation of cellulose and hemicelluloses (1.87 and 1.65-fold, respectively). The LMC-supplemented process was stable over a 90 d biogas production period. This work demonstrates the potential of LMC for enhancing biogas from lignocellulosic wastes.


Intestinal Ultrasonography in the Diagnosis and Management of Colonic Diverticular Disease.
Diverticula of the colon and their symptomatic manifestations, including acute diverticulitis (AD), are frequent complaints and the cause of an increasing burden of ambulatory visits, diagnostic procedures, and hospital admissions. Endoscopic and radiologic diagnostic procedures have a well-known role in the diagnosis and management of the disease, but recently intestinal ultrasonography has been proposed as a complementary tool in the diagnosis and follow-up of diverticular disease. This review shows the main sonographic features of diverticula and discusses the potential role of ultrasound in suggesting the presence of symptomatic uncomplicated diverticular disease of the colon. Moreover, the sonographic features of AD, diagnostic accuracy, advantages, and limitations of the technique will be discussed. We place special emphasis on the present role of intestinal ultrasonography in patients with suspected AD. Owing to its high sensitivity and high positive predictive value in assessing AD, intestinal ultrasound is currently suggested by some European national consensus guidelines as the first-line examination in this setting. In fact, to minimize false-negative findings and avoid unnecessary radiation exposure in patients with suspected AD, intestinal ultrasound might be used as the first-line examination in a sequential diagnostic strategy, followed by computed tomography only in the case of negative or inconclusive findings.


[A case of anal fistula cancer probably developing from intraluminal dissemination of rectal cancer].
A 63-year-old man with a history of anal fistula was admitted to our hospital because the anal pain didn't disappear after the operation. On digital examination, a hard mass measuring 3.0 cm in diameter was found at the anal canal. Biopsy of the mass showed moderately differentiated adenocarcinoma. Colonoscopy revealed another rectal cancer at 15 cm from anal verge. Biopsy of the tumor also showed moderately differentiated adenocarcinoma, resembling the anal canal tumor. Because the histological findings of both tumors were nearly identical, we considered that cancer cells from the rectal cancer had been implanted and developed the metastatic tumor in the anal fistula. The patient underwent anterior resection for the rectal cancer, and a local resection for the anal canal cancer. Immunohistochemical staining for Ki-67, p53, Muc2, CD10, CK-7, and CK-20 revealed similar patterns in both tumors. Additionally, genetic analysis for p53, K-ras, and MSI revealed similar patterns in both tumors. We may suggest from these results that cancer cells from the rectal cancer had been implanted and developed the metastatic tumor in the anal canal.


Molecular characterization of anaerobic microbial communities from benzene-degrading sediments under methanogenic conditions.
Anaerobic benzene degradation was confirmed in microbial communities enriched from Baltimore Harbor (Baltimore, MD) sediments under methanogenic conditions. Molecular characterization based on 16S rDNA gene sequences revealed that the strains in the communities were diversely affiliated with such phylogenetic branches as the Bacteroidetes, Euryarchaeota, Firmicutes, and Thermotogae phyla. Of interest was that the majority of the microbial populations detected in these cultures were closely related to the members of dechlorinating microbial communities. Further, some of those species were previously found in naphthalene- or phenanthrene-degrading methanogenic communities. Finally, this result could be used to design targeted isolation strategies for anaerobic benzene-degrading strains under methanogenic conditions.