Use of image analysis to understand enzyme stability in an aerated stirred reactor.
Efficient regeneration of NAD(P)+ cofactors is essential for large-scale application of alcohol dehydrogenases due to the high cost and chemical instability of these cofactors. NAD(P)+ can be regenerated effectively using NAD(P)H oxidases (NOXs) that require molecular oxygen as a cosubstrate. In large-scale biocatalytic processes, agitation and aeration are needed for sufficient oxygen transfer into the liquid phase, both of which have been shown to significantly increase the rate of enzyme deactivation. As such, the aim of this study was to identify the existence of a correlation between enzyme stability and gas-liquid interfacial area inside the bioreactor. This was done by measuring gas-liquid interfacial areas inside an aerated stirred reactor, using an in situ optical probe, and simultaneously measuring the kinetic stability of NOXs. Following enzyme incubation at various power inputs and gas-phase compositions, the residual activity was assessed and video samples were analyzed through an image processing algorithm. Enzyme deactivation was found to be proportional to an increase in interfacial area up to a certain limit, where power input appears to have a higher impact. Furthermore, the presence of oxygen increased enzyme deactivation rates at low interfacial areas. The areas were validated with defined glass beads and found to be in the range of those in large-scale bioreactors. Finally, a correlation between the enzyme half-life and specific interfacial area was obtained. Therefore, we conclude that the method developed in this contribution can help to predict the behavior of biocatalyst stability under industrially relevant conditions, concerning specific gas-liquid interfacial areas.


Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle.
The COSynch protocol has been used to synchronize ovulation and facilitate fixed-time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (</=11 mm). The reason for the presence of small follicles at the second GnRH (GnRH 2) is not clear. The objectives of this study were 1) to determine the effect of ovulatory response at the first GnRH (GnRH 1) on diameter and variation in diameter of the largest follicle at GnRH 2, and 2) to determine the effect of day of the cycle (stage of a follicular wave) on GnRH-induced luteinizing hormone (LH) release, and the resulting ovulatory response after GnRH 1 and 2. Two experiments used pubertal beef heifers synchronized to be on different days of the estrous cycle (d 2, 5, 10, 15, and 18 after estrus) in which a dominant follicle would or would not respond to GnRH 1. Ovulatory response to GnRH 1 did not affect size or variation in diameter of the largest follicle at GnRH 2 in Exp. 1 or 2. In Exp. 1, ovulatory response after GnRH 1 (0/14(a), 12/13(b), 4/13(ac), 9/13(bc), and 2/10(a) in the d 2, 5, 10, 15, and 18 groups; (a-c)P < 0.05) and GnRH 2 (13/14(a), 12/13(a), 12/13(a), 2/13(b), and 2/10(b) in the d 2, 5, 10, 15, and 18 groups, respectively; (a,b)P < 0.05) was affected by day of the cycle. In Exp. 2, day of the cycle also affected the proportion of heifers ovulating after GnRH 1 (0/7(a), 8/8(b), 0/6(a) 5/8(ab), and 5/8(ab) of the d 2, 5, 10, 15, and 18 heifers, respectively; (a-c)P < 0.05) and GnRH 2 (3/7(ab), 8/8(b), 5/6(b), 1/8(a), and 2/8(a) of the d 2, 5, 10, 15, and 18 heifers, respectively; (a,b)P < 0.05). In both experiments, heifers receiving GnRH 1 on d 15 and 18 had a greater (P < 0.05) occurrence of luteolysis before PGF(2alpha) injection and expression of estrus than heifers treated on d 2, 5, and 10. The GnRH-induced LH surge was of greatest magnitude in heifers receiving GnRH 1 on d 18 of the cycle followed by d 5, 15, 10, and 2 (9,054(b), 5,774(bc), 4,672(c), 2,548(c), and 915(d) arbitrary units; respectively; (a-d)P < 0.05). In summary, ovulatory response to GnRH 1 did not affect size of the dominant follicle at GnRH 2. Day of the cycle when GnRH 1 was delivered affected dominant follicle size at GnRH 2. Treatment with GnRH 1 in the earlier part of the estrous cycle (on or before d 10) increased the proportion of dominant follicles that were large enough to respond to GnRH 2 (>/=10 mm) and increased ovulatory response after GnRH 2.


[Internal medicine in the last 40 years].
Internal medicine in the last 40 years has known many eminent teachers at universities, such as Hijmans van den Bergh, Van Buchem, Borst, Mulder, Lindeboom, De Langen, Hulst, Jordan, Formijne, Major, Snapper, Groen and Querido, and others outside universities such as Stuyt, Van Hees, Eindhoven, Pannekoek, Schalm, Bruins Slot, Heeres, Stolte and Pompen. The main scientific platform on which the bequeathors assembled in the first few decades after World War II was the Algemene Ziektekundige Vereniging ('General Medical Association'), which met in Utrecht. Important medical steps forward in that period were the virtually complete eradication of tuberculosis in the Netherlands, the developing of vaccines against smallpox and polio, but also against the various types of virus hepatitis, and the progress in intensive care, transplantation and molecular biology. The Nederlandsche Internisten Vereeniging ('Dutch Association of Internists') has had its own scientific journal since 1958. Essentials in modern internal medicine are appropriate care and demonstrated usefulness of a treatment (evidence-based medicine); standards, values and ethics are core issues, as are quality control and cost control. Change is also in progress in the doctor-patient relationship, as manifested in the Wet Geneeskundige Behandelingsovereenkomst ('Medical Treatment Accord Act').


Octoclothepin enantiomers. A reinvestigation of their biochemical and pharmacological activity in relation to a new receptor-interaction model for dopamine D-2 receptor antagonists.
Octoclothepin (1) was resolved into its R and S enantiomers via the diastereomeric tartaric acid salts. The enantiomers were shown to be of high optical purity by 1H NMR with use of the chiral shift reagent (R)-(-)-2,2,2-trifluoro-1-(9-anthryl)ethanol. Pharmacological and biochemical testing confirmed that (S)-1 is the more potent dopamine (DA) D-2 antagonist both in vitro and in vivo, although the R enantiomer still has significant D-2 antagonistic activity. In contrast, both enantiomers were equally active in test models detecting activity at D-1 receptors, serotonin-2 (5-HT2) receptors and alpha 1 adrenoceptors. Contrary to a previous prediction, it was found that norepinephrine (NE) uptake inhibition was confined solely to the S enantiomer. Overall, (S)-1 has a "classical" neuroleptic profile, while the R enantiomer has a more "atypical" profile. These pharmacological profiles seem to be in agreement with the reported clinical profiles of the two enantiomers. A previous conformational study was revised in light of the biochemical test results with enantiomers of known optical purity. Their relative D-2 receptor affinity corresponded well with the calculated conformational energy difference between their "active conformations" deduced from a previously reported new D-2 receptor model. Also the high enantioselectivity of (S)-1 at the NE uptake site could be explained after a detailed conformational analysis showing strict requirements for the orientation of the piperazine lone-pair direction at the NE uptake site.


Long-term functional and radiological outcomes of cervical disc arthroplasty at a tertiary level spine center in India: A retrospective cohort analysis with minimum 2 years of follow-up.
Cervical disc arthroplasty (CDA) was developed for the treatment of cervical disc disease with the potential advantages of preservation of physiological motion at a discal level, thereby potentially reducing adjacent level stresses and degeneration, which were a known complication of anterior cervical arthrodesis. The objective of this study was the assessment of long-term functional and radiological outcomes overtime in all the patients who underwent CDA from 2011 to 2019 at our institute. Forty-eight patients who underwent CDA (2011-2019) with a minimum 2-year follow-up were retrospectively evaluated. The functional outcome included the Visual Analog Score (VAS) and Neck Disability Index (NDI). Radiographs were assessed for range of motion (ROM) at the index surgical level, presence of heterotopic ossification (HO), and adjacent segment degeneration. The mean follow-up was 5.79 ± 2.96 (2.16-11.75) years. Significant improvement (P < 0.05) was observed in the VAS (8.91 ± 2.52 [preoperative] to 0.89 ± 1.27 [follow-up]) and NDI (65.5% ±23.06% [preoperative] to 4.79 ± 3.87 [follow-up]) score. Motion at index level increased significantly from 5.53° preoperatively to 7.47°, and 92% of the implanted segments were still mobile (referring to the threshold of ROM > 3°). HOs are responsible for the fusion of 4/50 (8%) levels at the last follow-up. Distal and proximal adjacent disc degeneration occurred in 36% and 28% of patients, respectively. No migration of the implant was observed on the radiograph. Our study showed favorable clinical outcome of CDA with preservation of ROM at the index surgical level. CDA can be a promising alternative to anterior cervical arthrodesis when properly indicated.


[Ultrasonic assessment of the development of the uterus in childhood].
In 150 girls from the newborn period up to the 18. year the size of the uterus was measured by sonography. In all children the length, width, depth and volume of the uterus as well as its shape and other signs of development were determined. In the newborn period and in the following first months of life an estrogen stimulated uterus with the shape of a drop could be shown. The size of the uterus decreased till the end of the first year. The length of the uterus in the neonatal period was 4.0 +/- 0.5 cm, the volume was 3.6 +/- 1.9 cm3. In the following rest phase up to the age of 8 years the length of the uterus was 2.8 +/- 0.4 cm and the volume was 1.2 +/- 0.4 cm3. The shape of the uterus in this age group was tubular. One to two years before secondary pubertal signs could be shown, sonography already demonstrated the beginning of pubertal uterus growth. In the postpubertal period the uterus had the typical pear shape with a corpus/cervix-relation of 2:1. In the postpubertal period the length was 6.8 +/- 1 cm with a volume of 33 +/- 22 cm3. The knowledge of the normal developmental dates of the uterus in children is essential for the diagnosis of malformations and tumours of the pelvis, but also for the diagnosis of distorted puberty such as pubertas praecox, pubertas tarda, amenorrhea and growth retardation.


Modulation of acetylcholine-stimulated insulin release by glucose and gastric inhibitory polypeptide.
The interactions of glucose, acetylcholine and gastric inhibitory polypeptide in the regulation of insulin secretion were examined using the in situ perfused rat pancreas. Acetylcholine (1 x 10(-6) M) had no effect on the release of immunoreactive insulin in the presence of 2.2 x 10(-3) M glucose. However, in the presence of 4.4, 6.6 or 8.9 x 10(-3) M glucose, the same concentration of acetylcholine stimulated insulin secretion approximately fourfold. At the highest glucose concentration tested (17.8 x 10(-3) M), the stimulatory effect of acetylcholine on insulin release was less pronounced. The insulin response to acetylcholine was potentiated by the presence of gastric inhibitory polypeptide. This potentiation was most marked when the peptide was present at a concentration of 1 x 10(-9) M, whereas the effect of concomitant infusion of acetylcholine and 2 x 10(-10) M gastric inhibitory polypeptide was only slightly greater than additive. Both atropine (1 x 10(-6) M) and hexamethonium (1 x 10(-4) M) inhibited the insulin response to acetylcholine. However, neither of these cholinergic antagonists had a significant effect on glucose- or gastric inhibitory polypeptide-stimulated insulin secretion. These results demonstrate that the insulinotropic action of acetylcholine is glucose-dependent. A synergistic interaction may exist between acetylcholine and gastric inhibitory polypeptide at the beta-cell which does not involve glucose or gastric inhibitory polypeptide acting on cholinergic receptors in the pancreas. Cholinergic stimulation of insulin secretion in the perfused rat pancreas appears to be mediated by both muscarinic receptors on the beta-cell and nicotinic receptors, presumably on intrapancreatic ganglia.


Electrochemical and electrophysiological analysis of the effects of SK&F 104078 on prejunctional alpha 2-adrenoceptors.
SK&F 104078 is proposed to be a selective postjunctional alpha 2-adrenoceptor antagonist. We examined its effects on the electrically evoked release of endogenous noradrenaline (NA) from the rat tail artery and of ATP from the mouse vas deferens, as determined electrochemically or by extracellular recording, respectively. The alpha 2-adrenoceptor antagonist yohimbine (0.1 and 1 microM) increased NA release at both 2 and 20 Hz; the effect of yohimbine was further enhanced by the NA uptake blocker cocaine. SK&F 104078 (0.01-1 microM) did not change NA release at 2 or 20 Hz in the absence or presence of cocaine. NA release at 2 Hz was dose dependently depressed by the alpha 2-adrenoceptor agonist xylazine, an effect reversed by yohimbine (1 microM) but unaffected by SK&F 104078 (0.1 and 1 microM). Similarly, the inhibitory effect of xylazine on ATP release at 0.1 Hz was not influenced by SK&F 104078 (1 microM) but partially reversed by yohimbine (1 microM). The results show (i) that prejunctional alpha 2-adrenoceptors mediate autoinhibition of the release of endogenous NA and ATP from sympathetic nerves in the rat tail artery and in the mouse vas deferens, respectively, and (ii) that SK&F 104078 does not block these receptors in the concentration range 0.01-1 microM.


Aberrant expression of the costimulatory molecule CD40 ligand on monocytes from patients with systemic lupus erythematosus.
CD40 ligand (CD40L, CD154) is overexpressed on T and B cells in systemic lupus erythematosus (SLE). Monocytes have been shown to contribute to immune-mediated pathology in SLE and to express CD40L under certain conditions. Therefore, we studied CD40L expression on lupus monocytes ex vivo, as well as after activation in vitro. A highly significant sevenfold increase in the frequency of CD40L-expressing peripheral monocytes from 23 SLE patients, compared to 16 healthy individuals (mean percentage of CD40L(+)CD14(+) among CD14(+) cells, 11.7 versus 1.6), was found by flow cytometry. Increased CD40L expression on monocytes correlated significantly with disease activity, elevated gamma-globulin serum levels, as well as increased CD40L expression on T cells. CD40L expression by lupus monocytes was verified at both the mRNA and protein levels, while LPS stimulation was found to upregulate CD40L mRNA accumulation and surface protein expression. CD40L expression on activated lupus monocytes within anti-CD3-stimulated, mononuclear cell cultures was also enhanced compared to control-derived monocytes. These novel findings underscore the multiplicity of pathways through which monocytes may contribute to SLE pathology and suggest that T cell-independent CD40L-mediated cell to cell interactions may be also involved in humoral immune activation in SLE.


New insights into spontaneous pneumothorax: A review.
A spontaneous pneumothorax is a pneumothorax that does not arise from trauma or an iatrogenic cause. Although the traditional classification of either primary or secondary spontaneous pneumothorax based on the absence or presence of overt underlying lung disease is still widely used, it is now well recognised that primary spontaneous pneumothorax is associated with underlying pleuropulmonary disease. Current evidence indicates that computed tomography screening for underlying disease should be considered in patients who present with spontaneous pneumothorax. Recent evidence suggests that conservative management has similar recurrence rates, less complications and shorter hospital stay compared with invasive interventions, even in large primary spontaneous pneumothoraces of >50%. A more conservative approach which is based on clinical assessment rather than pneumothorax size can thus be followed during the acute management in selected stable patients. The purpose of this review is to revisit the aetiology of spontaneous pneumothorax, identify which patients should be investigated for secondary causes and to give an overview of the management strategies at initial presentation as well as secondary prevention.


VEGF induces phosphorylation of STAT3 through binding VEGFR2 in ovarian carcinoma cells in vitro.
VEGF plays a key role in ovarian carcinoma. Recent studies have shown that expressions of VEGF and its receptors were correlated with signal tranducer phosphorylation and activators of transcription 3(p-STAT3) in ovarian carcinoma. The aim of this study was to investigate the effects of STAT3 phosphorylation on VEGF signaling pathways in ovarian carcinoma cells. We selected an ovarian carcinoma cell line Caov-3 as a target cell that co-expressed VEGFR2 and p-STAT3. We detected expressions of p-STAT3 in Caov-3 induced by VEGF with different concentrations and for different effect times by immunocytochemistry and Western Blot. A concentration of 50 ng/ml VEGF was enough to increase phosphorylation of STAT3, and at 30 min, the p-STAT3 level reached the peak and showed nuclear translocation of p-STAT3 from the cytoplasm to the nucleus. These effects could be overcome by a small peptide (ATWLPPR) specific for VEGFR2. Taken together, VEGF-induced phosphorylation and nuclear translocation of STAT3 and ATWLPPR could effectively block the VEGF effects, suggesting that phosphorylation of STAT3 participates in VEGF signal transduction via VEGFR2 in ovarian carcinoma cells.


Histamine H2 receptor desensitization: involvement of a select array of G protein-coupled receptor kinases.
The histamine H2 receptor (H2r) belongs to the heptahelical receptor family; upon agonist binding, members of this family activate a G protein and the downstream effector adenylyl cyclase. Like other G protein-coupled receptors, exposure of H2r to agonists produces a desensitization of the response. The present study focused on the desensitization mechanism of this receptor. Using transiently transfected COS-7 cells expressing tagged-H2r, the desensitization induced by amthamine, characterized by decreased cAMP production, was studied. Results show that the receptor was rapidly desensitized with a t(1/2) = 0.49 +/- 0.01 min. Because of the rapid nature of H2r desensitization, receptor phosphorylation was examined as a likely mechanism for signal attenuation. H2r desensitization was not affected by protein kinases A and C (PKA and PKC) inhibitors but was remarkably reduced by Zn(2+), an inhibitor of G protein-coupled receptor kinases (GRKs). Cotransfection experiments using tagged H2r and different GRKs (2, 3, 5, or 6), demonstrated that GRK2 and GRK3 were the most potent in augmenting desensitization, causing a reduction in the maximal response to amthamine and a decrease of the t(1/2) for desensitization, whereas GRK5 and GRK6 did not affect the signaling. Receptor phosphorylation correlates with desensitization for each GRK studied, whereas phosphorylation that is dependent on protein kinases A and C seemed irrelevant in receptor signal termination. These results indicate that in H2r-transfected COS-7 cells, exposure to an agonist caused desensitization controlled by H2r phosphorylation via GRK2 and GRK3.


Conservation of old individual trees and small populations is integral to maintain species' genetic diversity of a historically fragmented woody perennial.
Historically fragmented and specialized habitats such as granite outcrops are understudied globally unique hot spots of plant evolution. In contrast to predictions based on mainstream population genetic theory, some granite outcrop plants appear to have persisted as very small populations despite prolonged geographic and genetic isolation. Eucalyptus caesia Benth. is a long-lived lignotuberous tree endemic with a naturally fragmented distribution on granite outcrops in south-western Australia. To quantify population to landscape-level genetic structure, we employed microsatellite genotyping at 14 loci of all plants in 18 stands of E. caesia. Sampled stands were characterized by low levels of genetic diversity, small absolute population sizes, localized clonality and strong fine-scale genetic subdivision. There was no significant relationship between population size and levels of heterozygosity. At the landscape scale, high levels of population genetic differentiation were most pronounced among representatives of the two subspecies in E. caesia as originally circumscribed. Past genetic interconnection was evident between some geographic neighbours separated by up to 20 km. Paradoxically, other pairs of neighbouring stands as little as 7 km apart were genetically distinct. There was no consistent pattern of isolation by distance across the 280 km range of E. caesia. Low levels of gene flow, together with strong drift within stands, provide some explanation of the patterns of genetic differentiation we observed. Individual genet longevity via the ability to repeatedly resprout and expand from a lignotuber may enhance the persistence of some woody perennial endemic plants despite small population size, minimal genetic interconnection and low heterozygosity.


External validation of a multifactorial normal tissue complication probability model for tube feeding dependence at 6 months after definitive radiotherapy for head and neck cancer.
The purpose of this study was to externally validate a previously published normal tissue complication probability (NTCP) model for tube feeding dependence at 6 months (TUBEM6) after completion of (chemo) radiotherapy. This study evaluated 122 head and neck cancer patients treated by definitive (chemo) radiotherapy. The closed testing procedure was used to select the appropriate method for updating the NTCP model. In this procedure, the likelihood ratio test was used to compare the updated model against the original model. Mean predicted NTCP was 12.2% (95% CI: 9.9%-14.5%) when using the original NTCP model for TUBEM6. TUBEM6 at our institute was 5.7% (95% CI: 1.8-9.6%) for the 122 patients evaluated. The test for the model revision against the original NTCP model was statistically significant (p = 0.032). The test for the model revision against the model adjusting intercept only was not statistically significant (p = 0.240). According to the closed testing procedure, the model required adjusting the intercept only. TUBEM6 at our institute was lower than that predicted by the original NTCP model. The closed testing procedure indicated that only an adjustment of the intercept was needed indicating the importance of external validation.


Radiation exposure induces cross-species temporal metabolic changes that are mitigated in mice by amifostine.
Exposure to acute, damaging radiation may occur through a variety of events from cancer therapy and industrial accidents to terrorist attacks and military actions. Our understanding of how to protect individuals and mitigate the effects of radiation injury or Acute Radiation Syndrome (ARS) is still limited. There are only a few Food and Drug Administration-approved therapies for ARS; whereas, amifostine is limited to treating low dose (0.7-6 Gy) radiation poisoning arising from cancer radiotherapy. An early intervention is critical to treat ARS, which necessitates identifying diagnostic biomarkers to quickly characterize radiation exposure. Towards this end, a multiplatform metabolomics study was performed to comprehensively characterize the temporal changes in metabolite levels from mice and non-human primate serum samples following γ-irradiation. The metabolomic signature of amifostine was also evaluated in mice as a model for radioprotection. The NMR and mass spectrometry metabolomics analysis identified 23 dysregulated pathways resulting from the radiation exposure. These metabolomic alterations exhibited distinct trajectories within glucose metabolism, phospholipid biosynthesis, and nucleotide metabolism. A return to baseline levels with amifostine treatment occurred for these pathways within a week of radiation exposure. Together, our data suggests a unique physiological change that is independent of radiation dose or species. Furthermore, a metabolic signature of radioprotection was observed through the use of amifostine prophylaxis of ARS.


What drives seed dispersal effectiveness?
Seed dispersal is a critical phase in plant reproduction and forest regeneration. In many systems, the vast majority of woody species rely on seed dispersal by fruit-eating animals. Animals differ in their size, movement patterns, seed handling, gut physiology, and many other factors that affect the number of seeds they disperse, the quality of treatment each individual seed receives, and consequently their relative contribution to plant fitness. The seed dispersal effectiveness framework (SDE) was developed to allow systematic and standardized quantification of these processes, offering a potential for understanding the large-scale dynamics of animal-plant interactions and the ecological and evolutionary consequences of animal behavior for plant reproductive success. Yet, despite its wide acceptance, the SDE framework has primarily been employed descriptively, almost always in the context of local systems. As such, the drivers of variation in SDE across systems and the relationship between its components remain unknown. We systematically searched studies that quantified endozoochorous SDE for multiple animal species dispersing one or more plant species in a given system and offered an integrative examination of the factors driving variation in SDE. Specifically, we addressed three main questions: (a) Is there a tradeoff between high dispersal quality and quantity? (b) Does animal body mass affect SDE or its main components? and (c) What drives more variation in SDE, seed dispersal quality, or quantity? We found that: (a) the relationship between quality and quantity is mediated by body size; (b) this is the result of differential relationships between body mass and the two components, while total SDE is unaffected by body mass; (c)neither quality nor quantity explain more variance in SDE globally. Our results also highlight the need for more standardized data to assess large-scale patterns in SDE.


The use of reflection as an assessment of practice. Can you mark learning contracts?
Learning contracts are used by many educational institutions as a means of assessing the practice of their students. However, there have been no studies into the reliability of marking learning contracts. A statistical analysis of the marks from the assi gnments of a practice module, which included a marked learning contract and a more traditional assessment strategy, namely, a clinical study, was made which measured the reliability of the marking process. This was undertaken in 3 ways: an analysis of th e clinical study as a measure of its reliability, secondly the learning contract marking itself was tested for reliability and finally a comparison between the two assessment methods. The statistical analysis was undertaken by means oft -test, intra-class coefficient calculation, weighted Kappa calculation, regression analysis and by a method outlined by Bland and Altman (1986, Lancet: 307-310) in which the set of means of two values is plotted against the differences. The clinical stud ies were found to be marked consistently, the maximum difference between the markers being only 8.3%, this value was then adopted as a measure of educational significance The inter-rater reliability of the learning contract marking was measured and was f ound to be not as high as the clinical studies. The t -test revealing a statistically significant (but not educationally) difference of 1.03%. The Bland and Altman plot revealed an maximum difference of 13.2%. However, a regression analysis revealed that 99.3% should fall within the 8.3% significance bandin g The clinical studies comparison with the learning contracts revealed that the difference as measured by t -test was significantly (not educationally) different at 2.3%. The Bland and Altman plot revealed that learning contracts may be marked up to 18.2% higher than the clinical studies but this may reflect the difference in what is being assessed by the two methods. However, it was noted that there was a greater agreement when the marks were higher. The validity of reflective learning contracts as an assessment tool has not been measured directly by this study, this would seem to be a natural next phase of analysis. This study supports the use of the assessment of learning contracts as a means of rigorous assessment of a student's practice.


Recurrent Pregnancy Loss and Concealed Long-QT Syndrome.
Background Recurrent pregnancy loss affects 1% to 2% of couples attempting childbirth. A large fraction of all cases remains idiopathic, which warrants research into monogenic causes of this distressing disorder. Methods and Results We investigated a nonconsanguineous Estonian family who had experienced 5 live births, intersected by 3 early pregnancy losses, and 6 fetal deaths, 3 of which occurred during the second trimester. No fetal malformations were described at the autopsies performed in 3 of 6 cases of fetal death. Parental and fetal chromosomal abnormalities (including submicroscopic) and maternal risk factors were excluded. Material for genetic testing was available from 4 miscarried cases (gestational weeks 11, 14, 17, and 18). Exome sequencing in 3 pregnancy losses and the mother identified no rare variants explicitly shared by the miscarried conceptuses. However, the mother and 2 pregnancy losses carried a heterozygous nonsynonymous variant, resulting in p.Val173Asp (rs199472695) in the ion channel gene KCNQ1. It is expressed not only in heart, where mutations cause type 1 long-QT syndrome, but also in other tissues, including uterus. The p.Val173Asp variant has been previously identified in a patient with type 1 long-QT syndrome, but not reported in the Genome Aggregation Database. With heterologous expression in CHO cells, our in vitro electrophysiologic studies indicated that the mutant slowly activating voltage-gated K+ channel (IKs) is dysfunctional. It showed reduced total activating and deactivating currents (P<0.01), with dramatically positive shift of voltage dependence of activation by ≈10 mV (P<0.05). Conclusions The current study uncovered concealed maternal type 1 long-QT syndrome as a potential novel cause behind recurrent fetal loss.


Effect of variations of corneal physiology on novel non-invasive intraocular pressure monitoring soft contact lens.
Glaucoma is the leading cause of irreversible blindness around the world. With its slow asymptomatic progression, there is an emphasis on early detection and frequent monitoring. A novel microfluidic contact lens has been established as a potential way to track the fluctuations of the intraocular pressure (IOP) which is a key indicator for diagnosing and monitoring glaucoma progression. The purpose of this article is to determine the effect of physiological variations of the eye on the performance of the microfluidic contact lens. Ultrasound biomicroscopy (UBM) was used to measure the central corneal thickness (CCT) and radius of corneal curvature (RCC) for a series of 16 fresh enucleated porcine eyes. The effect of these corneal anatomic features on device performance was then assessed by systematically adjusting intraocular pressure from 10 to 34 mmHg and monitoring the device indicator response. The performance of the microfluidic contact lens was determined by finding the amount the indicator fluid shifted in position as a result of 1 mmHg IOP increase. The relationship between IOP and indicator fluid was found to be linear for all eyes. The slope of the indicator fluid movement as a result of the IOP was evaluated against the CCT and RCC of each porcine eye. This yielded low correlation coefficients, 0.057 for CCT and 0.024 for RCC, meaning that these physiological differences showed no systematic impact on the measurements made with the contact lens.


Three-dimensional structure of the tryptophan synthase alpha 2 beta 2 multienzyme complex from Salmonella typhimurium.
The three-dimensional structure of the alpha 2 beta 2 complex of tryptophan synthase from Salmonella typhimurium has been determined by x-ray crystallography at 2.5 A resolution. The four polypeptide chains are arranged nearly linearly in an alpha beta beta alpha order forming a complex 150 A long. The overall polypeptide fold of the smaller alpha subunit, which cleaves indole glycerol phosphate, is that of an 8-fold alpha/beta barrel. The alpha subunit active site has been located by difference Fourier analysis of the binding of indole propanol phosphate, a competitive inhibitor of the alpha subunit and a close structural analog of the natural substrate. The larger pyridoxal phosphate-dependent beta subunit contains two domains of nearly equal size, folded into similar helix/sheet/helix structures. The binding site for the coenzyme pyridoxal phosphate lies deep within the interface between the two beta subunit domains. The active sites of neighboring alpha and beta subunits are separated by a distance of about 25 A. A tunnel with a diameter matching that of the intermediate substrate indole connects these active sites. The tunnel is believed to facilitate the diffusion of indole from its point of production in the alpha subunit active site to the site of tryptophan synthesis in the beta active site and thereby prevent its escape to the solvent during catalysis.


Effect of storage on physicochemical attributes of ice cream enriched with microencapsulated anthocyanins from black carrot.
The present study was conducted to investigate the effect of storage on quality attributes of microencapsulated black carrot anthocyanins-enriched ice cream. Purposely, black carrot anthocyanins were obtained using ethanolic extraction. Later on, extracts were acidified and microencapsulated with gum arabic and maltodextrin (1:1). Results showed that anthocyanin contents for T3 (9% microencapsulated anthocyanins powder-enriched ice cream) had highest anthocyanin contents in the range of 143.21 ± 1.14 mg/100 g. However, during the storage, it was revealed that there was a slight decline in constituents concentration reasoned to oxygen exposure and interaction with other food ingredients. Similarly, for total phenolic content, the highest amount was found in T3 as 545.38 ± 4.34 mg GAE/100 g. The quality attributes of prepared ice cream treatments were also found acceptable till the end of the study (60 days). Conclusively, the addition of microencapsulated anthocyanins powder in ice cream proved to stabilize black carrot anthocyanins and contributed positively to the sensory characteristics of ice cream.


Genome-Wide Analysis of Serine/Arginine-Rich Protein Family in Wheat and Brachypodium distachyon.
By regulating the pre-mRNA splicing of other genes and themselves, plant serine/arginine-rich (SR) proteins play important roles in development and in response to abiotic stresses. Presently, the functions of most plant SR protein genes remain unclear. Wheat (Triticum aestivum) and Brachypodium distachyon are closely related species. In this study, 40 TaSR and 18 BdSR proteins were identified respectively, and they were classified into seven subfamilies: SR, RS, SCL, RSZ, RS2Z, SC35, and SR45. Similar to Arabidopsis and rice SR protein genes, most TaSR and BdSR protein genes are expressed extensively. Surprisingly, real-time polymerase chain reaction (RT-PCR) analyses showed that no alternative splicing event was found in TaSR protein genes, and only six BdSR protein genes are alternatively spliced genes. The detected alternatively spliced BdSR protein genes and transcripts are much fewer than in Arabidopsis, rice, maize, and sorghum. In the promoter regions, 92 development-related, stress-related, and hormone-related cis-elements were detected, indicating their functions in development and in response to environmental stresses. Meanwhile, 19 TaSR and 16 BdSR proteins were predicted to interact with other SR proteins or non-SR proteins, implying that they are involved in other functions in addition to modulating pre-mRNA splicing as essential components of the spliceosome. These results lay a foundation for further analyses of these genes.


Nutritional Factors Affecting Mental Health.
Dietary intake and nutritional status of individuals are important factors affecting mental health and the development of psychiatric disorders. Majority of scientific evidence relating to mental health focuses on depression, cognitive function, and dementia, and limited evidence is available about other psychiatric disorders including schizophrenia. As life span of human being is increasing, the more the prevalence of mental disorders is, the more attention rises. Lists of suggested nutritional components that may be beneficial for mental health are omega-3 fatty acids, phospholipids, cholesterol, niacin, folate, vitamin B6, and vitamin B12. Saturated fat and simple sugar are considered detrimental to cognitive function. Evidence on the effect of cholesterol is conflicting; however, in general, blood cholesterol levels are negatively associated with the risk of depression. Collectively, the aims of this review are to introduce known nutritional factors for mental health, and to discuss recent issues of the nutritional impact on cognitive function and healthy brain aging.


Social modulation of androgen levels in male teleost fish.
Androgens are classically thought of as the sex steroids controlling male reproduction. However, in recent years evidence has accumulated showing that androgens can also be affected by the interactions between conspecifics, suggesting reciprocal interactions between androgens and behaviour. These results have been interpreted as an adaptation for individuals to adjust their agonistic motivation and to cope with changes in their social environment. Thus, male-male interactions would stimulate the production of androgens, and the levels of androgens would be a function of the stability of its social environment ['challenge hypothesis', Gen. Comp. Endocrinol. 56 (1984) 417]. Here the available data on social modulation of androgen levels in male teleosts are reviewed and some predictions of the challenge hypothesis are addressed using teleosts as a study model. We investigate the causal link between social status, territoriality and elevated androgen levels and the available evidence suggests that the social environment indeed modulates the endocrine axis of teleosts. The association between higher androgen levels and social rank emerges mainly in periods of social instability. As reported in the avian literature, in teleosts the trade-off between androgens and parental care is indicated by the fact that during the parental phase breeding males decreased their androgen levels. A comparison of androgen responsiveness between teleost species with different mating and parenting systems also reveals that parenting explains the variation observed in androgen responsiveness to a higher degree than the mating strategy. Finally, the adaptive value of social modulation of androgens and some of its evolutionary consequences are discussed.


[Current status and clinical course of hepatitis C virus in Korea].
The mortality due to chronic liver disease, including liver cirrhosis and hepatocellular carcinoma (HCC), ranks as one of the highest in Korea. The prevalence rates of hepatitis C virus (HCV) and hepatitis B virus (HBV) infections in the general Korean population are approximately 1 and 5%, respectively. Blood transfusion was the strongest risk factor for the transmission of HCV infection. Therefore, the evaluation of risk factors for HCV infection including blood transfusion, intravenous drug user, hemophilia, and hemodialysis, is important. The most prevalent HCV genotype is 1b followed by 2a. The annual incidence of HCC among HCV-related liver cirrhosis has been estimated at 5%, and approximately 12% of HCC is attributable to HCV and 68% to HBV in Korea. HCV infection is more closely associated with HCC in elderly patients than HBV-related HCC. Even though the prevalence of anti-HCV in Korea has been reduced and the risk of HCV transmission through blood transfusion has markedly decreased, public-health programs to prevent de novo infections should be developed. This review describes the HCV prevalence and risk factors among the general population, and the distribution of HCV genotypes as well as the clinical course of HCV in Korea.


Alternative splicing of a neural-specific Src mRNA (Src+) is a rapid and protein synthesis-independent response to neural induction in Xenopus laevis.
Alternative splicing is used by the src protooncogene to derive two gene products, pp60c-src and pp60+. pp60+ is expressed exclusively in neurons, whereas pp60c-src is expressed in many tissues. The specific role of the src+ gene product in neurons is not yet understood. Expression of src+ mRNA begins early in neural development and is restricted to the neural plate in Xenopus embryos. Using a reverse transcription/polymerase chain reaction assay, we demonstrate that expression of src+ mRNA is dependent on neural induction. Src+ mRNA is expressed in animal cap ectoderm which is cocultured with mesoderm tissue, but not in ectoderm cultured alone. The protein kinase C (PKC) activator 12-O-tetradecanoylphorbol-13-acetate (TPA) induces src+ mRNA expression in cultured stage 10+ animal cap ectoderm. This induction is rapid, with src+ mRNA being detected within 1 hr of TPA addition. Only dorsal ectoderm tissue is competent to express src+ mRNA upon TPA treatment, and stage 10+ ectoderm tissue becomes unresponsive to TPA after 4 hr in culture. TPA-dependent induction of src+ mRNA in ectoderm tissue is not blocked by cycloheximide and therefore is not dependent on protein synthesis.


Mental Task Classification Scheme Utilizing Correlation Coefficient Extracted from Interchannel Intrinsic Mode Function.
In view of recent increase of brain computer interface (BCI) based applications, the importance of efficient classification of various mental tasks has increased prodigiously nowadays. In order to obtain effective classification, efficient feature extraction scheme is necessary, for which, in the proposed method, the interchannel relationship among electroencephalogram (EEG) data is utilized. It is expected that the correlation obtained from different combination of channels will be different for different mental tasks, which can be exploited to extract distinctive feature. The empirical mode decomposition (EMD) technique is employed on a test EEG signal obtained from a channel, which provides a number of intrinsic mode functions (IMFs), and correlation coefficient is extracted from interchannel IMF data. Simultaneously, different statistical features are also obtained from each IMF. Finally, the feature matrix is formed utilizing interchannel correlation features and intrachannel statistical features of the selected IMFs of EEG signal. Different kernels of the support vector machine (SVM) classifier are used to carry out the classification task. An EEG dataset containing ten different combinations of five different mental tasks is utilized to demonstrate the classification performance and a very high level of accuracy is achieved by the proposed scheme compared to existing methods.


Autoantigens and Sjögren syndrome.
To review recent experimental evidence on the involvement of autoantigens and autoantibodies in the pathogenesis of organ-specific autoimmunity in Sjögren syndrome (SS). Among candidate autoantigens in SS, we investigated the role of alpha-fodrin in the lacrimal gland using a mouse model and a lacrimal gland cell line established in p53 knockout mice. We also tried to identify a novel organ-specific autoantigen by screening the human salivary gland cell line (HSG) against sera from SS patients. The specificity and sensitivity of this autoantigen to SS patient sera, its cellular localization, and the gene encoding the protein were analyzed. In addition to the previously identified 120-kd alpha-fodrin in the salivary gland, a shorter fragment was detected, indicating that there may be a distinct apoptosis-related protease that cleaves alpha-fodrin in the lacrimal gland. A novel salivary gland-specific autoantibody was detected in 50.9% of sera from SS patients. The antigen recognized by this antibody may be a 45-kd nucleus protein not recognized in its native form. The precise roles of autoantigens in organ-specific autoimmunity are still unclear, although accumulated evidence suggests that they may be associated with disease progression. Further studies of alpha-fodrin and the 45-kd antigen may contribute to the understanding of the pathogenesis of SS and may provide a new strategy for organ-specific therapy, such as vaccination with analogue peptides.


CUX1 Facilitates the Development of Oncogenic Properties Via Activating Wnt/β-Catenin Signaling Pathway in Glioma.
Background: Homeobox cut like 1 (CUX1), which often presents aberrated expression in many cancer cells, exerts a crucial role in tumorigenesis. Evidence describing CUX1 in gliomagenesis is scarce, and the effects of CUX1 on the Wnt/β-catenin pathway have not been reported. Our study aimed to explore the biological functions and molecular mechanisms involved in CUX1 activity in glioma. Methods: Datasets for bioinformatics analysis were obtained from the GEO, TCGA, CGGA, GTEX and CCLE databases. qRT-PCR, western blotting (WB), and immunohistochemistry (IHC) assays were used to investigate the expression patterns of CUX1 among glioma and brain tissues. CUX1 knockdown and overexpression vectors were transfected into glioma cell lines, the CCK-8, clone formation assay, wound healing, Transwell assay, and flow cytometry were performed to detect changes in cell viability, invasiveness, and the cell cycle. WB and immunofluorescence (IF) assays were used to explore changes in cell cycle-related and Wnt/β-catenin signaling protein levels. Results: Overexpression of CUX1 was identified in glioma tissues, and especially in glioblastoma (GBM), when compared to normal controls and correlated with poor prognosis. In comparison with untreated cells, TJ905 glioma cells overexpressing CUX1 showed higher proliferation and invasion abilities and S phase cell-cycle arrest, while the knockdown of CUX1 suppressed cell invasive ability and induced G1 phase arrest. Active Wnt/β-catenin signaling was enriched and clustered in a CUX1-associated GSEA/GSVA analysis. IF and WB assays indicated that CUX1 regulated the distribution of Axin2/β-catenin in glioma cells and regulated the expression of proteins downstream of the Wnt/β-catenin signaling pathway, suggesting that CUX1 served as an upstream positive regulator of the Wnt/β-catenin pathway. Finally, the knockdown of Axin2 or β-catenin could reverse the tumor-promoting effects caused by CUX1 overexpression, suggesting that CUX1 induced gliomagenesis and malignant phenotype by activating the Wnt/β-catenin signaling pathway. Conclusion: Our data suggested that the transcription factor CUX1 could be a novel therapeutic target for glioma with gene therapy.


Dorsal fractures of the triquetrum: MRI findings with an emphasis on dorsal carpal ligament injuries.
The objective of our study was to report the MRI findings in dorsal fractures of the triquetrum, with an emphasis on dorsal carpal ligament injuries. A total of 21 patients (16 men, five women; mean age, 41.9 years) with acute or subacute (≤ 6 weeks) dorsal triquetral fractures on radiography and MRI were included in this two-center retrospective study. MRI of the wrist was performed on 3-T units with transverse T1-weighted, coronal or transverse (or both) fat-suppressed T2weighted, transverse gadolinium-enhanced fat-suppressed T1-weighted turbo spin-echo, and 3D gadolinium-enhanced fat-suppressed T1-weighted gradient-recalled echo sequences. Three musculoskeletal radiologists evaluated the ulnar styloid process index (USPI) on radiographs and the following MRI features: fracture pattern (types 1-6), bone fragment size and displacement, bone marrow edema distribution, and dorsal carpal ligament tears. Eight type 1, one type 2, six type 3, five type 4, and one type 5 fractures were identified. These fractures were associated with 14 (66.7%), 17 (81.0%), and 16 (76.2%) tears of the dorsal radiocarpal, ulnotriquetral, and intercarpal ligaments, respectively. There was no correlation between bone marrow edema distribution and dorsal carpal ligament injuries (all p > 0.05). The mean (± SD) bone fragment volume and displacement were 205 ± 157 mm(3) and 1.0 ± 1.1 mm, respectively. The mean USPI was 0.21 ± 0.10. Dorsal fractures of the triquetrum are frequently associated with dorsal carpal ligament injuries. Bone marrow edema distribution is not correlated with these ligament tears.


The effects of temperature on the stability of a neuronal oscillator.
The crab Cancer borealis undergoes large daily fluctuations in environmental temperature (8-24°C) and must maintain appropriate neural function in the face of this perturbation. In the pyloric circuit of the crab stomatogastric ganglion, we pharmacologically isolated the pacemaker kernel (the AB and PD neurons) and characterized its behavior in response to temperature ramps from 7°C to 31°C. For moderate temperatures, the pacemaker displayed a frequency-temperature curve statistically indistinguishable from that of the intact circuit, and like the intact circuit maintained a constant duty cycle. At high temperatures (above 23°C), a variety of different behaviors were seen: in some preparations the pacemaker increased in frequency, in some it slowed, and in many preparations the pacemaker stopped oscillating ("crashed"). Furthermore, these crashes seemed to fall into two qualitatively different classes. Additionally, the animal-to-animal variability in frequency increased at high temperatures. We used a series of Morris-Lecar mathematical models to gain insight into these phenomena. The biophysical components of the final model have temperature sensitivities similar to those found in nature, and can crash via two qualitatively different mechanisms that resemble those observed experimentally. The crash type is determined by the precise parameters of the model at the reference temperature, 11°C, which could explain why some preparations seem to crash in one way and some in another. Furthermore, even models with very similar behavior at the reference temperature diverge greatly at high temperatures, resembling the experimental observations.


[Role of pharmacists in disaster medicine: required knowledge and skills].
Immediately after the Great East Japan Earthquake of March 2011, Okayama University dispatched a medical assistance team based on the request of Iwate Prefecture. The first team was followed by 12 medical teams. I was one of the members of the fourth and fifth medical teams sent to Rikuzen-takata and Ofunato for a week beginning March 16th to support medical relief operations as a pharmacist during the sub-acute phase of the disaster. As a member of the team at the temporary clinic in Rikuzen-takata, pharmacists such as myself required physical assessment skills to perform related tasks, along with expertise in drug dispensing and consultation. In my next medical team, which headed the pneumonia unit at Oofunato Hospital, I played a critical role in the effective use of medicine reserved/provided for disasters, including antibiotics. Throughout the relief operations, strong clinical reasoning and decision making, as well as good teamwork, proved vital, especially in emergency situations. For future community medical systems, emergency/disaster medicine should be included in pharmacy education. The School of Pharmacy at Okayama University will establish emergency medicine program in the next school year, in cooperation with the Medical, Dental and Health Care Departments.


Binding characteristics of the calcium channel antagonist [3H]-nitrendipine in tissues from ethanol-dependent rats.
Male Sprague-Dawley rats were made physically dependent on ethanol by exposure to ethanol vapour in inhalation chambers for 6-10 days. Animals were sacrificed immediately after removal from the chambers. After sacrifice, membrane preparations from cerebral cortex, heart, vas deferens and thigh skeletal muscle were prepared and studied to determine the characteristics of [3H]-nitrendipine binding. In membranes from cerebral cortex, the number of binding sites (Bmax) was increased 40% in preparations from ethanol-dependent rats compared to controls. Similar results were obtained in heart (Bmax increased 125%), vas deferens (Bmax increased 50%) and skeletal muscle (Bmax increased 33%). Binding affinity (Kd) in all these tissues was unchanged. Membranes from cortex and heart were studied more extensively. Displacement studies using other dihydropyridines and the effects of ionic composition on [3H]-nitrendipine binding provided no evidence to suppose that the binding sites induced by the development of ethanol dependence were from a population different from those in control tissues. The results suggest a generalised 'up-regulation' of the dihydropyridine-sensitive subclass of voltage operated calcium channels in excitable tissues from ethanol-dependent rats.


Liver cyst with biliary communication successfully treated with laparoscopic deroofing: a case report.
A 71-year-old Japanese woman complained of right upper abdominal fullness and pain. Computed tomography revealed a huge cyst in the right lobe of the liver, measuring 16 cm in diameter. She underwent laparoscopic deroofing of the liver cyst. On operation, needle aspiration of the cyst yielded clear serous fluid without any bile contamination. However, after the cyst was deroofed with laparoscopic coagulating shears, bile leakage was recognized from a tiny orifice in the cyst cavity. A catheter was inserted via the orifice for cholangiography, which demonstrated a communication with the biliary tract. The orifice was easily closed with a laparoscopic suturing device. Operation time was 5 hours and 30 minutes, and blood loss was 300 grams. Pathological examination of the liver cyst was consistent with a simple cyst. The postoperative course was uneventful, and the patient has had no recurrence to date at 13 months. Laparoscopic deroofing is a recommended treatment for a liver cyst even in the presence of cystobiliary communication.


Vitreous Hemorrhage following High-Altitude Retinopathy.
To report a case of high-altitude retinopathy with vitreous hemorrhage. An apparently healthy 29-year-old boy presented with a history of floater and slight diminution of vision in the left eye after climbing the mountain 4760 meters high. The visual acuity at presentation was 20/20 in the right eye and 20/30 in the left eye. Anterior segment findings of both eyes were unremarkable. Fundus evaluation revealed bilaterally dilated major retinal veins. The right eye revealed clear, quiet vitreous, healthy macula, and pink and smooth optic disc. There was fresh vitreous hemorrhage confined just one disc diameter away at the superior and inferior part of the optic nerve in the left eye. The macula and optic nerve head of this eye grossly looked normal. Complete blood count, haemoglobin, ESR, CRP, sugar, renal function test, lipid panel, and serology for HIV, HCV, VDRL, and HBsAg were normal. The Mantoux test and chest X-ray also revealed normal findings. Physician consultation did not reveal other abnormalities. On the 3rd week of follow-up, his vision was 20/20 in both eyes. Fundus examination revealed clear vitreous in both eyes though some venous dilation and tortuosities were still evident in the left eye. A macular OCT scan showed almost normal-appearing fundus. Vitreous hemorrhage following climbing high altitude can be one of the manifestations of high-altitude retinopathy.


Effects of ethinyl estradiol plus desogestrel on premenstrual symptoms in Iranian women.
Marvelon®, a combined oral contraceptive, contains 30 μg ethinyl estradiol (EE) and 150 μg desogestrel (DE), and has been shown to be a well-tolerated and effective combination that provides high contraceptive reliability and good cycle control. However, its efficacy has not been yet evaluated among Iranian women. Thus, the study aimed to determine the effect of oral contraceptive pill on treating premenstrual symptoms and on various parameters associated with well-being and health in a sample of Iranian. This clinical trial (before- after) study was performed at the family-planning clinic of the centers under the supervision of Tehran University of Medical Sciences on sixty-one women. The study protocol was approved by the Ethics Committee of Tehran University of Medical Sciences and all participants received a 21/7-day regimen of oral contraceptive containing 150 μg desogestrel (DE) and 30 μg ethinyl estradiol (EE) for six cycles. Efficacy parameters included changes in premenstrual symptoms were also assessed. Clinical data was collected by calendar of premenstrual experiences (COPE) at baseline and treatment cycles 1,2, 3 and 6. Clinical variables were measured including low-density lipoprotein (LDL), high-density lipoprotein (HDL) and triglyceride levels for two timing periods (baseline and last visit). Linear mixed model analyses were used to analyze differences in changes of the four factors of premenstrual syndrome (PMS), weight and blood pressure during these timing periods. The mean age of the women was 28.52 (SD=6.75) years. Participants on average had been pregnant 1.13 (SD=1.16) times. The linear mixed model analyses indicated that premenstrual syndrome symptoms reduced significantly over time (P<0.05). Marvelon® showed no significant effect on reducing LDL and HDL levels, and participant's weights were also stable during five-time assessments (P>0.05). A combined oral contraceptive containing ethinyl estradiol and desogestrel has a positive effect on women's health and reduces premenstrual symptoms.


Bone mineral density status in urolithiasis patients with vitamin D inadequacy followed at a tertiary stone centre.
We assessed abnormalities in bone mineral density (BMD) and the risk of hip and major osteoporotic fractures in urolithiasis patients with vitamin D inadequacy (VDI) followed at a tertiary stone centre. Stone-free patients with VDI were invited to undergo dual-energy x-ray absorptiometry (DXA) scans to assess for BMD abnormalities at the femoral neck and lumbar spine. The World Health Organization's validated Fracture Risk Assessment Tool (FRAX) was used to calculate the risk of hip and major osteoporotic fractures within 10 years. Patients with primary hyperparathyroidism or hypercalcemia were excluded. In total, 50 consecutive patients were included between June 2011 and August 2012, including 26 (52%) males. The median age was 51 years and the median 25-hydroxyl vitamin D (25[OH] D) was 18.8 ng/mL. Thirty patients (60%) had abnormal T-scores on DXA studies. This decreased to 22 (44%) when age-matched Z-scores were used; 36% had osteopenia and 8% had osteoporosis. Femoral neck and lumbar spines were affected in 24% and 32% of patients, respectively. Recurrent stone-formers had significantly lower BMD when compared with first-time stone formers. Median serum 25(OH)D was comparable between patients with normal and abnormal DXA scans (18.6 vs. 18.8 ng/mL; p = 0.91). Five patients (10%) were at high risk (≥3%) of hip fractures within 10 years. A high prevalence of abnormal DXA scans was found in urolithiasis patients with VDI, including 5 patients (10%) at high risk of hip fractures. Future studies need to assess the economic impact of obtaining DXA scans on urolithiasis patients with VDI, especially in recurrent stone-formers.


Long-term stability of soft tissues following alveolar ridge preservation: 10-year results of a prospective study around nonsubmerged implants.
The purpose of this study was to evaluate the long-term clinical outcomes around implants placed in sites previously augmented with demineralized bovine bone mineral with 10% collagen (Bio-Oss Collagen, Geistlich). In this prospective study, 36 consecutive, healthy patients, in need of a single-tooth extraction (incisors, canines, and premolars) and implant replacement, were included. After tooth extraction, Bio-Oss Collagen was inserted in the socket and covered either with a double layer of collagen membrane (test) or with a few drops of a flowable polylactide polymer (control). Following a healing period of 4 to 6 months, a single nonsubmerged implant surgery was performed. After cementation of a single ceramic crown, patients were asked to follow an individualized supportive periodontal therapy program. Clinical and radiographic data were obtained after prosthesis delivery (baseline) and at the 10-year follow-up visit. At the 10-year examination, two patients were lost to follow-up. All implants demonstrated healthy peri-implant soft tissues as documented by standard parameters (full-mouth plaque score, full-mouth bleeding score, local bleeding on probing) in both groups. Mean soft tissue recession (REC) was 0.39 ± 0.54 mm for the test group and 0.50 ± 0.33 mm for the control, with no significant difference between the two groups. The results of this prospective study confirmed the long-term stability of the peri-implant marginal soft tissues supported by regenerated bone by means of the described technique using Bio-Oss Collagen. If the patient is properly followed throughout time, the risk for mucosal recession is low, with < 1 mm of mean REC after 10 years.


Meaning-making in the immune system.
Meaning-making is the process by which a system responds to an indeterminate signal. This article focuses on meaning-making in living systems. It proposes several guidelines for studying the process of meaning-making in living systems in general, and in the immune system in particular. Drawing on a general framework for studying meaning-making in living systems, I suggest three basic organizing concepts for studying meaning-making-variability of the signal, context markers, and transgradience. Those concepts present a radical alternative to the information-processing approach that governs biological research and may shed new light on biological processes.


Alpha and beta diversity jointly drive the aboveground biomass in temperate and tropical forests.
Changes in biodiversity often affect ecosystem functioning. However, most previous biodiversity and ecosystem functioning (BEF) studies have generally been limited to very small spatial grains. Thus, knowledge regarding the biodiversity-ecosystem functioning relationships across spatial scales is lacking. Moreover, the multiscale nature of biodiversity, and specifically β diversity (i.e., spatial heterogeneity in species composition) was still largely missing in BEF studies. Here, using the vegetation and functional trait data collected from four 6-ha forest dynamics plots (FDPs) in temperate and tropical forests in China, we examine the scale-dependent relationships between tree diversity and the aboveground biomass (AGB), as well as the roles of species spatial heterogeneity in determining the AGB. In tropical forests, the effect of species richness on AGB decreased with spatial grains, while functional dominance played a stronger role at larger spatial grains. In temperate forests, positive relationship between diversity and AGB occurred at all spatial grains, especially on smaller scales. In both temperate and tropical forests, β diversity was positively correlated with AGB, but weaker than α diversity in determining AGB. Overall, complementarity and selection hypothesis play dominant role in determining AGB in temperate and tropical forests, respectively. The roles of these underlying mechanisms are more pronounced with increasing spatial scales. β diversity, a hitherto underexplored facet of biodiversity, is likely to increase ecosystem functions by species spatial turnover and should not be neglected in BEF explorations. Our findings have practical implications for forest management and demonstrate that biotic heterogeneity plays an important positive role in ecosystem functioning.


Aberrant induction of regulatory activity of CD4+CD25+ T cells by dendritic cells in HIV-infected persons with amebic liver abscess.
To know why HIV-1-infected persons are particularly susceptible to amebic liver abscess (ALA), we investigated the role of CD4CD25 T cells in the susceptibility of HIV-1-infected persons to this disease. Herein we show, in early stage HIV-1-infected subjects, that CD4 T-cell responses to Entamoeba histolytica antigen (EhAg) were selectively impaired, especially in those with ALA. EhAg-specific CD4 T-cell responses were normalized by depletion of CD4CD25 cells or by addition of anti-cytotoxic T lymphocyte antigen 4 (CTLA4) antibody. Regulatory activity of CD4CD25 T cells to suppress the EhAg-specific CD4 T-cell response could be induced by EhAg-primed dendritic cells (DCs) in HIV-1-infected subjects, especially in those with ALA, but not in healthy controls. Exogenous Tat-incubated DCs derived from HIV-negative subjects also could upregulate CTLA4 expression on autologous CD4CD25 T cells and selectively suppress the EhAg-specific CD4 T-cell response. The results imply an interaction of the two pathogens: HIV-1, perhaps through the effect of Tat on DCs, may upregulate EhAg-specific regulatory T-cell activity to suppress T-cell response to E. histolytica, thus increasing the susceptibility to invasive amebiasis in even early-stage HIV-1-infected persons.


IN VITRO EFFICACY OF EXTRACTS FROM PLANTS USED BY SMALL-HOLDER FARMERS IN THE TREATMENT OF DERMATOPHILOSIS IN CATTLE.
Bovine dermatophilosis, an important skin disease of cattle caused by Dermatophilus congolensis, negatively impacts the livelihoods of small-holder farmers in Zimbabwe. This impact is through, morbidity, loss of draught animal power, costs incurred to manage the disease, losses associated with devalued damaged hides and the resultant culling of some of the affected cattle. Due to the inaccessibility of conventional drugs to manage bovine dermatophilosis, farmers have been reported to use local medicinal plants to manage the disease. The aim of the study was to evaluate the in vitro antimicrobial activities of three plants that small-holder farmers in Zimbabwe used to manage bovine dermatophilosis. Dried plant materials were ground into powder and extracted individually using, water, 80 % acetone and 80 % methanol. The antimicrobial properties of the plants were evaluated against two Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and one Gram-positive (Staphylococcus aureus) reference bacterial strains. They were further evaluated against a field isolate of Dermatophilus congolensis. The assays used were the disc diffusion, minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). Acetone and methanol extracts had superior inhibitory activities than did those of water. Pterocarpus angolensis DC extracts had better inhibitory properties with absolute MIC values of 0.156 - 5 mg/ml, Cissus Quadrangularis L had MIC values in the range 0.156 - 5 mg/ml while that of Catunaregam spinosa Thunb, Terveng was 0.156 - 10 mg/ml. Dermatophilus congolensis was more sensitive to Pterocarpus angolensis DC average MIC = 0.63 mg/ml than to Cissus quadrangularis L average MIC = 1.25 mg/ml and Catunaregam. spinosa Thunb, Terveng average MIC = 2.08 mg/ml. These results suggest the potential antibacterial activities of extracts of the three plants and hence farmers are, in a way, justified in using the plants. Better results (lower MIC) could be obtained by extracting and evaluating pure active compounds of the plants.


The DNA-binding and transcription-activation abilities of p53 are necessary but not sufficient for its antiproliferation function.
Normal p53 protein suppresses cell proliferation and ras oncogene-induced cell transformation. Missense mutations in the middle conserved conformational domain of p53 decrease its antiproliferation function. In this work, we studied the requirement of the NH2- and COOH-terminal regions of p53 in its antiproliferation function using two independent assays, growth of chronic myelogenous leukemia K562 cells on methylcellulose semisolid medium and ras oncogene-induced focus formation of rat fibroblast cells (Rat-1). We found that deletion of 80 or 159 amino acids from the NH2-terminus and deletion of 67 amino acids from the COOH-terminus of p53 drastically reduced the antiproliferation function of p53. However, the COOH-terminal deletion mutant is capable of binding to a p53 DNA-binding element, p53CON (GGACATGCCCGGGCATGTCC), and of activating p53CON-mediated transcription. These results suggest that p53' abilities to bind p53CON and activate transcription are not sufficient for its antiproliferation function and that p53CON-regulated genes may not be growth suppressive.


Experimental small bowel obstruction in chick embryos: Effects on the developing enteric nervous system.
After surgical repair of congenital small bowel atresias, intestinal motility disorders often are observed. These may be caused by changes in the enteric nervous system (ENS) secondary to obstruction. To assess these changes, small bowel atresias were induced experimentally in chick embryos. On day 11, the small intestines of 90 chicken embryos were ligated microsurgically in ovo. Breeding of the eggs was continued until day 19. The small bowel was removed, fixed, and embedded for silver-staining, semithin serial sections, and transmission electron microscopy. Additionally, acetyl-cholinesterase (AChE)-staining was performed. Normal chick embryos of the same age served as controls. Macroscopically, experimentally induced small bowel atresias had the same characteristics as human newborns. Microscopically, the wall structure was preserved; however, the ENS differed markedly from controls. Both proximal and distal to the obstruction, the submucosal plexus was almost completely absent, whereas the myententeric plexus was diminished only in the proximal dilated blind pouch. The axonal net was disrupted additionally. Ganglion cells of the myenteric plexus in the proximal segment were arranged in longitudinal clusters of densely packed cells. In the distal segment ganglion cells formed round clusters. The cells of Cajal, which normally surround the myenteric ganglia, were absent in the proximal and distal segments. In our experiments, structural changes in the ENS could be observed secondary to experimentally induced small bowel atresias in the chick. Because of the lack of ischemia in this model, the main cause of these ENS changes seems to be the dilatation oft the proximal gut. Dilatations are common features in intestinal atresias, anorectal malformations, and Hirschsprung's disease. Our observations, thus, explain motility disorders after the surgical repair of these diseases.


Reduced uptake of liposomal idarubicin in the perfused rat heart.
Although idarubicin is one of the most important anthracyclines and liposomal formulations seemed less cardiotoxic than free drug formulations, there are no definite data on cardiac uptake of liposomal encapsulated idarubicin. This study has been designed to determine uptake and negative inotropic action of liposomal idarubicin in the single-pass isolated perfused rat heart. Idarubicin-bearing liposomes, composed of 1,2-distearoyl-sn-glicero-phosphocholine, 1,2-distearoyl-sn-glicero-phosphoethanolamine-N-[poly(ethylene glycol)2000], and cholesterol were administered as a 10-min constant infusion of 1 mg followed by a 70-min washout period. Outflow concentration and left ventricular-developed pressure were measured and compared with data of free idarubicin observed previously under the same experimental conditions. Liposomal encapsulation significantly reduced cardiac uptake of idarubicin to about 15% and extensively diminished its negative inotropic action to less than 5% of the values observed for free idarubicin.


Device-Related Thrombus After Left Atrial Appendage Occlusion With the Amulet Device.
Left atrial appendage occlusion (LAAO) is increasingly used for stroke prevention in patients with atrial fibrillation who are considered unsuitable for a lifelong oral anticoagulant regimen. Recently, a single-centre study reported device-related thrombus formation in 16.7% of patients treated with the second-generation Amulet device (St. Jude Medical, St. Paul, MN, USA), presenting a potential major safety concern. As "real-world" data on device-related thrombus formation following LAAO with the Amulet occluder are scarce, we aimed to evaluate this outcome in a retrospective registry. Clinical and tranosesophageal echocardiography data after LAAO with the Amulet in consecutive patients from three centres were collated. Among 38 patients (mean age 75.8 years), mean (standard deviation) CHA2DS2-VASc and HAS-BLED scores were 4.4 (1.2) and 3.4 (0.9), respectively. All patients underwent successful device placement without procedure-related adverse events. The antithrombotic regimen at discharge consisted of dual antiplatelet therapy (DAPT) in 27 patients (71.1%), single antiplatelet therapy in 10 patients (26.3%), and no antithrombotic therapy in one patient (2.6%). Device-related thrombus was observed in one patient (2.6%) despite DAPT regimen. The outcome of this patient was uncomplicated after adjustment of oral anticoagulant therapy. No patients presented with a thromboembolic event following LAAO during a mean (standard deviation) follow-up of 15 (5) months. In this retrospective study, device-related thrombus formation with the second-generation Amulet device was rare and occurred at a rate similar to that of the previous device. Importantly, no patient experienced a device-related thromboembolic event during follow-up. Larger real-life studies are required to confirm the safety profile of this increasingly used device.


Epigenetic modifications and their relation to caste and sex determination and adult division of labor in the stingless bee Melipona scutellaris.
Stingless bees of the genus Melipona, have long been considered an enigmatic case among social insects for their mode of caste determination, where in addition to larval food type and quantity, the genotype also has a saying, as proposed over 50 years ago by Warwick E. Kerr. Several attempts have since tried to test his Mendelian two-loci/two-alleles segregation hypothesis, but only recently a single gene crucial for sex determination in bees was evidenced to be sex-specifically spliced and also caste-specifically expressed in a Melipona species. Since alternative splicing is frequently associated with epigenetic marks, and the epigenetic status plays a major role in setting the caste phenotype in the honey bee, we investigated here epigenetic chromatin modification in the stingless bee Melipona scutellaris. We used an ELISA-based methodology to quantify global methylation status and western blot assays to reveal histone modifications. The results evidenced DNA methylation/demethylation events in larvae and pupae, and significant differences in histone methylation and phosphorylation between newly emerged adult queens and workers. The epigenetic dynamics seen in this stingless bee species represent a new facet in the caste determination process in Melipona bees and suggest a possible mechanism that is likely to link a genotype component to the larval diet and adult social behavior of these bees.


Prion infection of mouse brain reveals multiple new upregulated genes involved in neuroinflammation or signal transduction.
Gliosis is often a preclinical pathological finding in neurodegenerative diseases, including prion diseases, but the mechanisms facilitating gliosis and neuronal damage in these diseases are not understood. To expand our knowledge of the neuroinflammatory response in prion diseases, we assessed the expression of key genes and proteins involved in the inflammatory response and signal transduction in mouse brain at various times after scrapie infection. In brains of scrapie-infected mice at pre- and postclinical stages, we identified 15 previously unreported differentially expressed genes related to inflammation or activation of the STAT signal transduction pathway. Levels for the majority of differentially expressed genes increased with time postinfection. In quantitative immunoblotting experiments of STAT proteins, STAT1α, phosphorylated-STAT1α (pSTAT1α), and pSTAT3 were increased between 94 and 131 days postinfection (p.i.) in brains of mice infected with strain 22L. Furthermore, a select group of STAT-associated genes was increased preclinically during scrapie infection, suggesting early activation of the STAT signal transduction pathway. Comparison of inflammatory markers between mice infected with scrapie strains 22L and RML indicated that the inflammatory responses and gene expression profiles in the brains were strikingly similar, even though these scrapie strains infect different brain regions. The endogenous interleukin-1 receptor antagonist (IL-1Ra), an inflammatory marker, was newly identified as increasing preclinically in our model and therefore might influence scrapie pathogenesis in vivo. However, in IL-1Ra-deficient or overexpressor transgenic mice inoculated with scrapie, neither loss nor overexpression of IL-1Ra demonstrated any observable effect on gliosis, protease-resistant prion protein (PrPres) formation, disease tempo, pathology, or expression of the inflammatory genes analyzed. Prion infection leads to PrPres deposition, gliosis, and neuroinflammation in the central nervous system before signs of clinical illness. Using a scrapie mouse model of prion disease to assess various time points postinoculation, we identified 15 unreported genes that were increased in the brains of scrapie-infected mice and were associated with inflammation and/or JAK-STAT activation. Comparison of mice infected with two scrapie strains (22L and RML), which have dissimilar neuropathologies, indicated that the inflammatory responses and gene expression profiles in the brains were similar. Genes that increased prior to clinical signs might be involved in controlling scrapie infection or in facilitating damage to host tissues. We tested the possible role of the endogenous IL-1Ra, which was increased at 70 days p.i. In scrapie-infected mice deficient in or overexpressing IL-1Ra, there was no observable effect on gliosis, PrPres formation, disease tempo, pathology, or expression of inflammatory genes analyzed.


Rehabilitation for the home care patient with COPD.
Assessment techniques and treatment strategies, such as medical management, oxygen management, chest physical therapy, and pulmonary rehabilitation, for patients with chronic obstructive pulmonary disease (COPD) are discussed. Clinically appropriate, evidence-based rehabilitation programs for home care may help to reduce care costs, reduce the impact of this chronic disease on patients with COPD, and prevent emergent care and rehospitalization.


Dioxotungsten 1,2-benzenedithiolate complex stabilized by NH...S hydrogen bonds.
Novel dioxo-tungsten(VI) bis(1,2-benzenedithiolate) complexes with neighboring amide groups, as models for tungsten enzymes, (NEt4)2[W(VI)O2{1,2-S(2)-3,6-(RCONH)2C6H2}2] (R = CH3, t-Bu), were designed and synthesized. The presence of the NH...S hydrogen bond was confirmed through IR spectrometry and X-ray crystallographic analysis. In the W(VI)O2 complexes, the NH...S hydrogen bond trans to the oxo ligand is stronger than that cis to oxo. On the basis of comparisons with [W(VI)O2(1,2-S2C6H4)2](2-), the NH...S hydrogen bond positively shifted the W(VI)/W(V) redox potentials and depressed the reduction by benzoin or triphenylphosphine. These results suggest that the NH...S hydrogen bond stabilizes the oxo ligand through trans influence and regulates O-atom transfer in tungsten and molybdenum enzymes.


Acoustic profiling in a complexly social species, the American crow: caws encode information on caller sex, identity, and behavioural context.
Previous research on inter-individual variation in the calls of corvids has largely been restricted to single call types, such as alarm or contact calls, and has rarely considered the effects of age on call structure. This study explores structural variation in a contextually diverse set of "caw" calls of the American crow (Corvus brachyrhynchos), including alarm, foraging recruitment and territorial calls, and searches for structural features that may be associated with behavioural context and caller sex, age, and identity. Automated pitch detection algorithms are used to generate 23 pitch-related and spectral parameters for a collection of caws from 18 wild, marked crows. Using principal component analysis and mixed models, we identify independent axes of acoustic variation associated with behavioural context and with caller sex, respectively. We also have moderate success predicting caller sex and identity from call structure. However, we do not find significant acoustic variation with respect to caller age.


HIV behind bars: human immunodeficiency virus cluster analysis and drug resistance in a reference correctional unit from southern Brazil.
People deprived of liberty in prisons are at higher risk of infection by the human immunodeficiency virus (HIV) due to their increased exposure through intravenous drug use, unprotected sexual activity, tattooing in prison and blood exposure in fights and rebellions. Yet, the contribution of intramural HIV transmission to the epidemic is scarcely known, especially in low- and middle-income settings. In this study, we surveyed 1,667 inmates incarcerated at Presídio Central de Porto Alegre, located in southern Brazil, for HIV infection and molecular characterization. The HIV seroprevalence was 6.6% (110/1,667). Further analyses were carried out on 40 HIV-seropositive inmates to assess HIV transmission clusters and drug resistance within the facility with the use of molecular and phylogenetic techniques. The molecular epidemiology of HIV-1 subtypes observed was similar to the one reported for the general population in southern Brazil, with the predominance of HIV-1 subtypes C, B, CRF31_BC and unique BC recombinants. In particular, the high rate (24%) of URF_BC found here may reflect multiple exposures of the population investigated to HIV infection. We failed to find HIV-infected inmates sharing transmission clusters with each other. Importantly, the analysis of HIV-1 pol genomic fragments evidenced high rates of HIV primary and secondary (acquired) drug resistance and an alarming proportion of virologic failure among patients under treatment, unveiling suboptimal access to antiretroviral therapy (ARV), low ARV adherence and dissemination of drug resistant HIV strains in primary infections. Our results call for immediate actions of public authority to implement preventive measures, serological screening and, for HIV-seropositive subjects, clinical and treatment follow-up in order to control HIV infection and limit the spread of drug resistance strains in Brazilian prisons.


Monoclonal antibodies recognizing tumor-associated antigen of human ovarian mucinous cystadenocarcinomas.
Spleen cells from BALB/c mice immunized with human ovarian cystadenocarcinoma extract were fused with the mouse myeloma cell line P3/NSI/1-Ag 4 in the presence of polyethylene glycol (Mr 4000). Of the 46 hybrids obtained, four secreted antibodies preferentially reactive to the immunizing ovarian tumor extract. Two of these hybrids, which showed no reaction with normal controls, were selected for cloning by the limiting dilution method. The numerous clones obtained from each hybrid were screened against a panel of five ovarian tumor extracts, pooled normal ovary extracts, and pooled normal human sera. One clone from each hybrid that showed specificity for ovarian mucinous cystadenocarcinomas was recloned to assure monoclonality and to establish a permanent hybridoma cell line. The antibodies secreted by these cell lines were of IgG1 subclass with kappa light chains. These antibody-producing hybridomas were selected for further analysis of the antibody specificity by a solid-phase radioimmunoassay and quantitative absorption tests. The monoclonal antibodies recognized an antigenic determinant present only in mucinous cystadenocarcinomas of the ovary. These did not react with any other gynecological or nongynecological tumor thus far tested. The antigen was not demonstrable in any normal adult tissues tested. Among fetal tissues examined, only fetal intestine extract showed a positive reaction. The antigenic determinant recognized by these monoclonal antibodies was unrelated to carcinoembryonic antigen, normal glycoprotein, normal human serum components, or human ABO blood group materials. These antibodies, which have relatively high affinity and can be produced in large amounts, will be useful for the isolation and immunochemical characterization of this antigen. The purified antigen and the specific antibodies could be then combined in a sensitive radioimmunoassay for the early detection of the antigen in the sera and body fluids of patients with ovarian mucinous cystadenocarcinomas.


Sleep in older adults and in subjects with dementia.
The neuronal structures for the regulation of sleep and wakefulness are located in the brain. This complex network is vulnerable to numerous factors, most importantly neurodegenerative diseases and drugs. The macrostructure and microstructure of sleep change with age. These changes are more pronounced in subjects with dementia. Sleep disorders in subjects with dementia may be independent of dementia or caused by dementia. Furthermore, epidemiological studies reveal that sleep disorders per se may induce dementia by reduction of cerebral clearance of beta-amyloids. The population attributable risk (PAR) of sleep disturbances to the incidence of dementia is estimated to be about 15%; therefore, management of sleep disturbances in older adults and subjects with dementia gives the opportunity of an impact on incidence and course of dementia. Sleep history should be taken from each individual and obvious sleep disturbances, especially sleep apnea, should be managed according to current guidelines. Future studies that concern the incidence and the management of dementia must take into account sleep and sleep disturbances.


A phylogenetic assessment of the meadow lizard Darevskia praticola (Eversmann, 1834) from Iran.
The meadow lizard Darevskia praticola (Eversmann, 1834) displays a wide distribution range from the Western Balkans and Caucasus to the Hyrcanian forest in Iran. Based on previous studies, four subspecies have been described (D. p. praticola, D. p. pontica, D. p. hyrcanica and D. p. loriensis), but molecular evidence suggests two main phylogenetic lineages, Balkan and Caucasus. In this study we incorporated geographic records and analysed new samples from Alborz Mountains (Gilan Province, Iran), exploring their phylogenetic affinities using two mitochondrial markers (Cyt b and ND4). For the first time, Iranian samples are included in a molecular phylogeny. Our results meet the previous suggestions regarding the presence of a nuclear mitochondrial DNA fragment (numt) in Cyt b, causing misleading phylogenetic inferences since it renders the species paraphyletic. In contrast, the ND4 genealogy groups the species into a monophyletic clade, as expected. Samples attributed to D. p. hyrcanica (Azerbaijan and Iran) and D. p. loriensis (Armenia) are settled in a single clade with minute genetic distance, suggesting synonymisation.


Neuromelanin-sensitive MRI correlates of cognitive and motor function in Parkinson's disease with freezing of gait.
Substantia nigra pars compacta (SNc) and locus coeruleus (LC) are neuromelanin-rich nuclei implicated in diverse cognitive and motor processes in normal brain function and disease. However, their roles in aging and neurodegenerative disease mechanisms have remained unclear due to a lack of tools to study them in vivo. Preclinical and post-mortem human investigations indicate that the relationship between tissue neuromelanin content and neurodegeneration is complex. Neuromelanin exhibits both neuroprotective and cytotoxic characteristics, and tissue neuromelanin content varies across the lifespan, exhibiting an inverted U-shaped relationship with age. Neuromelanin-sensitive MRI (NM-MRI) is an emerging modality that allows measurement of neuromelanin-associated contrast in SNc and LC in humans. NM-MRI robustly detects disease effects in these structures in neurodegenerative and psychiatric conditions, including Parkinson's disease (PD). Previous NM-MRI studies of PD have largely focused on detecting disease group effects, but few studies have reported NM-MRI correlations with phenotype. Because neuromelanin dynamics are complex, we hypothesize that they are best interpreted in the context of both disease stage and aging, with neuromelanin loss correlating with symptoms most clearly in advanced stages where neuromelanin loss and neurodegeneration are coupled. We tested this hypothesis using NM-MRI to measure SNc and LC volumes in healthy older adult control individuals and in PD patients with and without freezing of gait (FOG), a severe and disabling PD symptom. We assessed for group differences and correlations between NM-MRI measures and aging, cognition and motor deficits. SNc volume was significantly decreased in PD with FOG compared to controls. SNc volume correlated significantly with motor symptoms and cognitive measures in PD with FOG, but not in PD without FOG. SNc volume correlated significantly with aging in PD. When PD patients were stratified by disease duration, SNc volume correlated with aging, cognition, and motor deficits only in PD with disease duration >5 years. We conclude that in severe or advanced PD, identified by either FOG or disease duration >5 years, the observed correlations between SNc volume and aging, cognition, and motor function may reflect the coupling of neuromelanin loss with neurodegeneration and the associated emergence of a linear relationship between NM-MRI measures and phenotype.


Genetic Variant of Kalirin Gene Is Associated with Ischemic Stroke in a Chinese Han Population.
Ischemic stroke is a complex disorder resulting from the interplay of genetic and environmental factors. Previous studies showed that kalirin gene variations were associated with cardiovascular disease. However, the association between this gene and ischemic stroke was unknown. We performed this study to confirm if kalirin gene variation was associated with ischemic stroke. We enrolled 385 ischemic stroke patients and 362 controls from China. Three SNPs of kalirin gene were genotyped by means of ligase detection reaction-PCR method. Data was processed with SPSS and SHEsis platform. SNP rs7620580 (dominant model: OR = 1.590, p = 0.002 and adjusted OR = 1.662, p = 0.014; additive model: OR = 1.490, p = 0.002 and adjusted OR = 1.636, p = 0.005; recessive model: OR = 2.686, p = 0.039) and SNP rs1708303 (dominant model: OR = 1.523, p = 0.007 and adjusted OR = 1.604, p = 0.028; additive model: OR = 1.438, p = 0.01 and adjusted OR = 1.476, p = 0.039) were associated with ischemic stroke. The GG genotype and G allele of SNP rs7620580 were associated with a risk for ischemic stroke with an adjusted OR of 3.195 and an OR of 1.446, respectively. Haplotype analysis revealed that A-T-G,G-T-A, and A-T-A haplotypes were associated with ischemic stroke. Our results provide evidence that kalirin gene variations were associated with ischemic stroke in the Chinese Han population.


The role of social support networks for rural hospital nurses: supporting and sustaining the rural nursing work force.
This survey study describes the workplace social support networks of rural hospital nurses in one geographic region of Colorado. This rural sample was compared with data from a sample of urban hospital nurses in Colorado. Social support networks influence nurses' satisfaction and retention. Nursing recruitment and retention is especially critical in rural areas where it takes healthcare facilities 60% longer than urban facilities to fill nursing vacancies. Little is known about rural nurses' social support networks. The Social Network Questionnaire (SNQ) was mailed to all nurses within one rural region of Colorado. Descriptive, correlational and comparative statistics were used to evaluate the structure and function of these nurses' networks. The nurses' networks were predominantly peer-based, but managers provided significant functional supports. The types of support provided by managers depended on the managers' placement in the nurses' networks. Some managers provided only performance feedback, and other managers offered physical assistance and emotional support. Rural nurses expected more guidance from management than did their urban counterparts. This sample of nurses clearly delineated the types of leadership styles most beneficial to them. Communication of roles and expectations between staff and management can enhance nursing satisfaction in the workplace setting.


Dyslexia: two priorities for the 1980s.
Within the past decade significant advances have been made in articulating the unique neuropsychological and cognitive processes in dyslexic children. The brain mapping procedures and postmortum studies are especially important in this regard. However, two important issues remain largely unaddressed. First, to date, no recognized nosology for dyslexia or subtypes of dyslexia exist. Second, the functional neurolinguistic system involved in reading and severe reading failure is, as of yet, still not completely understood in regard to subtypes of dyslexia. This article discusses these two areas of priority and proposes a framework for addressing these areas of concern.


Exploring Anti-Type 2 Diabetes Mellitus Mechanism of Gegen Qinlian Decoction by Network Pharmacology and Experimental Validation.
Gegen Qinlian Decoction (GGQL) has been employed to treat type 2 diabetes mellitus (T2DM) in the clinical practice of traditional Chinese medicine. However, the underlying mechanism of GGQL in the treatment of T2DM remains unknown. This study was aimed at exploring the pharmacological mechanisms of GGQL against T2DM via network pharmacology analysis combined with experimental validation. The effective components of GGQL were screened, and the target was predicted by using traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP). The candidate targets of GGQL were predicted by network pharmacological analysis, and crucial targets were chosen by the protein-protein interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses were performed to predict the core targets and pathways of GGQL against T2DM. Then, T2DM mice were induced by a high-fat diet combined with streptozotocin. The model and GGQL groups were given normal saline and GGQL aqueous solution (10 and 20 g/kg/d) intragastric administration, respectively, for 8 weeks. The mice in the GGQLT groups were administered with GGQLT at 10 and 20 g/kg/d, respectively. The pathological changes in liver tissues were observed by hematoxylin-eosin staining. The protein expression of TNF-α and NF-κB was verified by western blotting. A total of 204 common targets of GGQL for the treatment of T2DM were obtained from 140 active ingredients and 212 potential targets of T2DM. GO and KEGG enrichment analysis involved 119 signaling pathways, mainly in inflammatory TNF signaling pathways. Animal experiments showed that GGQL significantly reduced the serum levels of body mass, fasting blood glucose, fasting insulin, HOMA-IR, TNF-α, and IL-17. The liver pathological section showed that GGQL could improve the vacuolar degeneration and lipid deposition in the liver of T2DM mice. Mechanistically, GGQL downregulated the mRNA expression of TNF-α and NF-κB. This study demonstrated that GGQL may exert antidiabetic effects against T2DM by suppressing TNF-α signaling pathway activation, thus providing a basis for its potential use in clinical practice and further study in treating T2DM.


Zernike phase contrast cryo-electron tomography.
Cryo-tomography in the electron microscope is unique in its ability to provide high-resolution, three-dimensional structural information about cells, organelles and macromolecules in a nearly native, frozen-hydrated state. However, the phase-contrast imaging method used in conventional cryo-electron tomography fails to faithfully represent the full range of structural features in such specimens. Only certain features are recorded with adequate contrast, and overall contrast is low. The recently developed Zernike phase contrast method has the potential to solve this problem, and here we apply it for the first time to cryo-electron tomography. The new method has uniform transfer characteristics for a wide range of spatial frequencies, leading to improved overall signal-to-noise ratio and raising the prospects of higher resolution and quantitative representation of specimen densities in the reconstructed tomograms.