Impact evaluation of the efficacy of different doses of vitamin D supplementation during pregnancy on pregnancy and birth outcomes: a randomised, controlled, dose comparison trial in Pakistan.
Vitamin D deficiency during pregnancy is a public health problem in Pakistan and is prevalent among most women of reproductive age in the country. Vitamin D supplementation during pregnancy is suggested to prevent adverse pregnancy outcomes and vitamin D deficiency in both the mother and her newborn. We conducted a double-blinded, randomised controlled trial in Karachi, Pakistan to evaluate the effect of different doses of vitamin D supplementation during pregnancy on biochemical markers (serum 25(OH)D, calcium, phosphorus and alkaline phosphatase) in women and neonates, and on pregnancy and birth outcomes (gestational diabetes, pre-eclampsia, low birth weight, preterm births and stillbirths). Pregnant women (N=350) in their first trimester were recruited and randomised to three treatment groups of vitamin D supplementation: 4000 IU/day (group A, n=120), 2000 IU/day (group B, n=115) or 400 IU/day (group C, n=115). Women and their newborn in group A had the lowest vitamin D deficiency at endline (endline: 75.9%; neonatal: 64.9%), followed by group B (endline: 84.9%; neonatal: 73.7%) and then the control group (endline: 90.2%; neonatal: 91.8%). Vitamin D deficiency was significantly lower in group A than in group C (p=0.006) among women at endline and lower in both groups A and B than in the control group (p=0.001) in neonates. Within groups, serum 25(OH)D was significantly higher between baseline and endline in group A and between maternal baseline and neonatal levels in groups A and B. Participant serum 25(OH)D levels at the end of the trial were positively correlated with those in intervention group A (4000 IU/day) (β=4.16, 95% CI 1.6 to 6.7, p=0.002), with food group consumption (β=0.95, 95% CI 0.01 to 1.89, p=0.047) and with baseline levels of serum 25(OH)D (β=0.43, 95% CI 0.29 to 0.58, p<0.0001). The evidence provided in our study indicates that vitamin D supplementation of 4000 IU/day was more effective in reducing vitamin D deficiency among pregnant women and in improving serum 25(OH)D levels in mothers and their neonates compared with 2000 IU/day and 400 IU/day. Trial registration number NCT02215213.


Breast tumors from CHEK2 1100delC-mutation carriers: genomic landscape and clinical implications.
Checkpoint kinase 2 (CHEK2) is a moderate penetrance breast cancer risk gene, whose truncating mutation 1100delC increases the risk about twofold. We investigated gene copy-number aberrations and gene-expression profiles that are typical for breast tumors of CHEK2 1100delC-mutation carriers. In total, 126 breast tumor tissue specimens including 32 samples from patients carrying CHEK2 1100delC were studied in array-comparative genomic hybridization (aCGH) and gene-expression (GEX) experiments. After dimensionality reduction with CGHregions R package, CHEK2 1100delC-associated regions in the aCGH data were detected by the Wilcoxon rank-sum test. The linear model was fitted to GEX data with R package limma. Genes whose expression levels were associated with CHEK2 1100delC mutation were detected by the bayesian method. We discovered four lost and three gained CHEK2 1100delC-related loci. These include losses of 1p13.3-31.3, 8p21.1-2, 8p23.1-2, and 17p12-13.1 as well as gains of 12q13.11-3, 16p13.3, and 19p13.3. Twenty-eight genes located on these regions showed differential expression between CHEK2 1100delC and other tumors, nominating them as candidates for CHEK2 1100delC-associated tumor-progression drivers. These included CLCA1 on 1p22 as well as CALCOCO1, SBEM, and LRP1 on 12q13. Altogether, 188 genes were differentially expressed between CHEK2 1100delC and other tumors. Of these, 144 had elevated and 44, reduced expression levels.Our results suggest the WNT pathway as a driver of tumorigenesis in breast tumors of CHEK2 1100delC-mutation carriers and a role for the olfactory receptor protein family in cancer progression. Differences in the expression of the 188 CHEK2 1100delC-associated genes divided breast tumor samples from three independent datasets into two groups that differed in their relapse-free survival time. We have shown that copy-number aberrations of certain genomic regions are associated with CHEK2 mutation 1100delC. On these regions, we identified potential drivers of CHEK2 1100delC-associated tumorigenesis, whose role in cancer progression is worth investigating. Furthermore, poorer survival related to the CHEK2 1100delC gene-expression signature highlights pathways that are likely to have a role in the development of metastatic disease in carriers of the CHEK2 1100delC mutation.


Underexpression of hsa-miR-449 family and their promoter hypermethylation in infertile men: A case-control study.
Post-transcriptional microRNAs (miRNAs) have a impotrant pattern in the spermatogenesis process. Study of the expression and methylation of hsa-miR-449 family in sperm samples of infertile men. In this case-control study, we recruited 74 infertile men (with asthenozoospermia, teratozoospermia, asthenoteratozoospermia, and oligoasthenoteratozoospermia) and 30 control samles. Methylation-specific PCR (MSP) method was used for methylation evaluation of hsa-miR-449 a, b, c promoter. By Real time PCR (qRT-PCR) method,we showed downregulation of hsa-miR-449 a, b, c in the sperm samples of infertile men and compared it to their fertile counterparts. There was significant underexperssion, in hsa-miR-449-b in oligoasthenoteratospermic samples (p = 0.0001, F = 2.9). About the methylation pattern, infertile men showed high frequency of methylation in the promoter of hsa-miR-449 a, b, c in comparison to controls (60.8% vs 23.3%), the highest amount of methylation was observed in oligoasthenoteratospermia samples (81.2%). In this study, low expression and high methylation of hsa-miR-449-b were observed in infertile men in compared to control samples, which can be one of the causes of defective spermatogenesis.


Polygenic scores for social science: Clarification, consensus, and controversy.
In this response, I focus on clarifying my arguments, highlighting consensus, and addressing competing views about the utility of polygenic scores (PGSs) for social science. I also discuss an assortment of expansions to my arguments and suggest alternative approaches. I conclude by reiterating the need for caution and appropriate scientific skepticism.


Assessing the Predictive Power of Relative Binding Free Energy Calculations for Test Cases Involving Displacement of Binding Site Water Molecules.
Improved sampling methodologies, more accurate force fields, and access to longer simulation time scales have led to an increased application of Relative Binding Free Energy (RBFE) calculations in drug discovery projects. In order to assess the strengths and limitations of such tools, adequate benchmark sets are required that challenge the methodology in certain well-defined aspects. We applied Free Energy Perturbation (FEP) calculations to six congeneric ligand pairs taken from the literature, in which addition of a functional group resulted in the displacement of buried binding site water molecules and compared the calculated relative binding free energies with the experimental ones. We started the perturbations from different initial solvation states and registered large inconsistencies (large hysteresis) between the calculated values. We furthermore applied a Grand Canonical Monte Carlo (GCMC) solvent sampling step prior to the FEP calculation that led to a smaller hysteresis for the simulations. By applying a hydration site analysis to the trajectories of the end-states of the perturbation, we could point out that the low accuracy of the predictions as well as the high dependence of the prediction on the chosen initial state is likely caused by the trapping of binding site water molecules and/or insufficient solvation of buried cavities that are formed upon completion of the perturbation. This work highlights that RBFE calculations can suffer from slow solvent exchange of buried parts of the binding sites with the bulk.


Pathological versus protective functions of IL-22 in airway inflammation are regulated by IL-17A.
IL-22 has both proinflammatory and tissue-protective properties depending on the context in which it is expressed. However, the factors that influence the functional outcomes of IL-22 expression remain poorly defined. We demonstrate that after administration of a high dose of bleomycin that induces acute tissue damage and airway inflammation and is lethal to wild-type (WT) mice, Th17 cell-derived IL-22 and IL-17A are expressed in the lung. Bleomycin-induced disease was ameliorated in Il22-/- mice or after anti-IL-22 monoclonal antibody (mAb) treatment of WT mice, indicating a proinflammatory/pathological role for IL-22 in airway inflammation. However, despite increased bleomycin-induced IL-22 production, Il17a-/- mice were protected from airway inflammation, suggesting that IL-17A may regulate the expression and/or proinflammatory properties of IL-22. Consistent with this, IL-17A inhibited IL-22 production by Th17 cells, and exogenous administration of IL-22 could only promote airway inflammation in vivo by acting in synergy with IL-17A. Anti-IL-22 mAb was delivered to Il17a-/- mice and was found to exacerbate bleomycin-induced airway inflammation, indicating that IL-22 is tissue protective in the absence of IL-17A. Finally, in an in vitro culture system, IL-22 administration protected airway epithelial cells from bleomycin-induced apoptosis, and this protection was reversed after coadministration of IL-17A. These data identify that IL-17A can regulate the expression, proinflammatory properties, and tissue-protective functions of IL-22, and indicate that the presence or absence of IL-17A governs the proinflammatory versus tissue-protective properties of IL-22 in a model of airway damage and inflammation.


Assessment of acetylcholinesterase and butyrylcholinesterase activities in blood plasma of agriculture workers.
Cholinesterase determination indicates whether the person has been under pesticide exposure is not. It is recommended that the worker's cholinesterase level should be assessed for workers at a pesticide applied region. Hence, cholinesterase activities in blood samples of agricultural workers exposed to vegetables and grape cultivation with age matched, unexposed workers, who never had any exposure to pesticides, were estimated. The detailed occupational history and lifestyle characters were obtained by questionnaire. Cholinesterase activity was determined by the method of Ellman as modified by Chambers and Chambers. AChE was ranging from 1.65 to 3.54μmoles/min/ml in exposed subjects where as it was ranged from 2.22 to 3.51μmoles/min/ml in control subjects. BChE activity was ranging from 0.16 to 5.2μmoles/min/ml among exposed subjects, where as it was ranged from 2.19 to 5.06μmoles/min/ml in control subjects. The results showed statistically significant reduction in enzyme activities (AChE 14%; BChE 56%) among exposed subjects. It was concluded that the reduction in cholinesterase activity may lead to varieties of effects. Hence it is compulsory to use protective gadgets during pesticide spray. Further a continuous biomonitoring study is recommended to assess pesticide exposure.


Bim gene dosage is critical in modulating nephron progenitor survival in the absence of microRNAs during kidney development.
Low nephron endowment at birth has been associated with an increased risk for developing hypertension and chronic kidney disease. We demonstrated in an earlier study that conditional deletion of the microRNA (miRNA)-processing enzyme Dicer from nephron progenitors results in premature depletion of the progenitors and increased expression of the proapoptotic protein Bim (also known as Bcl-2L11). In this study, we generated a compound mouse model with conditional deletion of both Dicer and Bim, to determine the biologic significance of increased Bim expression in Dicer-deficient nephron progenitors. The loss of Bim partially restored the number of nephron progenitors and improved nephron formation. The number of progenitors undergoing apoptosis was significantly reduced in kidneys with loss of a single allele, or both alleles, of Bim compared to mutant kidneys. Furthermore, 2 miRNAs expressed in nephron progenitors (miR-17 and miR-106b) regulated Bim levels in vitro and in vivo Together, these data suggest that miRNA-mediated regulation of Bim controls nephron progenitor survival during nephrogenesis, as one potential means of regulating nephron endowment.-Cerqueira, D. M., Bodnar, A. J., Phua, Y. L., Freer, R., Hemker, S. L., Walensky, L. D., Hukriede, N. A., Ho, J. Bim gene dosage is critical in modulating nephron progenitor survival in the absence of microRNAs during kidney development.


Radiological and hormonal responses of functioning pituitary adenomas after gamma knife radiosurgery.
In this study, we examined patients with functioning pituitary adenoma that underwent Gamma Knife radiosurgery (GKS). In particular, we assessed the effects of GKS on the growth and endocrinological response of the functioning pituitary adenoma. Forty-two cases of functioning pituitary adenoma treated with GKS were analyzed. The mean follow-up duration was 42.5 months (range 6 - 98), and the mean tumor volume was 1.4 cm(3) (range 0.1 - 3.8). Multiple isocenters, ranging from 1 to 6 in number (mean 2.7), were used. The tumor margin was covered by an isodose ranging from 50 to 90%. The margin dose was 18 to 40 Gy (mean 28.5) and the maximum dose varied from 35 to 80 Gy (mean 54.1). Tumor growth was controlled in 96.9% of the cases and tumor shrinkage occurred in 40.6% of the cases. Hormonal response was observed in 35 of the 42 (83.3%) patients after GKS, with a mean duration of 6.8 months. Sixteen of the 42 (38.1%) patients showed hormonal normalization, with a mean duration of 21 months. In our multivariate analysis, high integral dosage (p=0.005) and maximum dosage (p=0.001) correlated significantly with hormonal normalization. For patients with functioning pituitary adenoma, GKS can be effective in controlling tumor growth and inducing hormonal normalization, especially if patients are reluctant to undergo surgical resection, or are not able to undergo microsurgery under general anesthesia. It appears that early hormonal normalization can be induced by high maximum dosage (at least 50 Gy) and broad coverage of the target volume within the isodose curve, while keeping the maximum dose to the visual pathways below 9 Gy.


Screening for post-traumatic stress disorder following childbirth using the Peritraumatic Distress Inventory.
Post-traumatic stress disorder (PTSD) following traumatic childbirth may undermine maternal and infant health, but screening for maternal childbirth-related PTSD (CB-PTSD) remains lacking. Acute emotional distress in response to a traumatic experience strongly associates with PTSD. The Peritraumatic Distress Inventory (PDI) assesses acute distress in non-postpartum individuals, but its use to classify women likely to endorse CB-PTSD is unknown. 3039 women provided information about their mental health and childbirth experience. They completed the PDI regarding their recent childbirth event, and a PTSD symptom screen to determine CB-PTSD. We employed Exploratory Graph Analysis and bootstrapping to reveal the PDI's factorial structure and optimal cutoff value for CB-PTSD classification. Factor analysis revealed two strongly correlated stable factors based on a modified version of the PDI: (1) negative emotions and (2) bodily arousal and threat appraisal. A score of 15+ on the modified PDI produced high sensitivity and specificity: 88 % with a positive CB-PTSD screen in the first postpartum months and 93 % with a negative screen. In this cross-sectional study, the PDI was administered at different timepoints postpartum. Future work should examine the PDI's predictive utility for screening women as closely as possible to the time of childbirth, and establish clinical cutoffs in populations after complicated deliveries. Brief self-report screening concerning a woman's emotional reactions to childbirth using our modified PDI tool can detect those likely to endorse CB-PTSD in the early postpartum. This may serve as the initial step of managing symptoms to ultimately prevent chronic manifestations.


An evaluation of the conductivity profile in the somatosensory barrel cortex of Wistar rats.
Microelectrode arrays used to record local field potentials from the brain are being built with increasingly more spatial resolution, ranging from the initially developed laminar arrays to those with planar and three-dimensional (3D) formats. In parallel with such development in recording techniques, current source density (CSD) analyses have recently been expanded up to the continuous-3D form. Unfortunately, the effect of the conductivity profile on the CSD analysis performed with contemporary microelectrode arrays has not yet been evaluated and most of the studies assumed it was homogeneous and isotropic. In this study, we measured the conductivity profile in the somatosensory barrel cortex of Wistar rats. To that end, we combined multisite electrophysiological data recorded with a homemade assembly of silicon-based probes and a nonlinear least-squares algorithm that implicitly assumed that the cerebral cortex of rodents could be locally approximated as a layered anisotropic spherical volume conductor. The eccentricity of the six cortical layers in the somatosensory barrel cortex was evaluated from postmortem histological images. We provided evidence for the local spherical character of the entire barrels field, with concentric cortical layers. We found significant laminar dependencies in the conductivity values with radial/tangential anisotropies. These results were in agreement with the layer-dependent orientations of myelinated axons, but hardly related to densities of cells. Finally, we demonstrated through simulations that ignoring the real conductivity profile in the somatosensory barrel cortex of rats caused considerable errors in the CSD reconstruction, with pronounced effects on the continuous-3D form and charge-unbalanced CSD. We concluded that the conductivity profile must be included in future developments of CSD analysis, especially for rodents.


The effects of low-color-temperature dual-primary-color light-emitting diodes on three kinds of retinal cells.
Long-term illumination of the retina with blue-light-excited phosphor-converted light-emitting diodes (LEDs) may result in decreased retinal function, even if the levels of blue light emitted are low. New low-color-temperature dual-primary-color LEDs have been developed that are composed of only two LED chips: a red chip and a yellow chip. These LEDs are expected to become a new type of healthy lighting source because they do not emit blue light, they lack phosphor, and they solve the problem of low efficiency encountered with phosphor-converted low-color-temperature LEDs. Many studies have indicated that these new low-color-temperature LEDs are likely to have therapeutic effects. However, the biological safety of these LEDs needs to be explored before the therapeutic effects are explored. Therefore, this experiment was conducted to investigate the effects of the new low-color-temperature LEDs and fluorescent white LEDs on three types of retinal cells. We observed that the viability and numbers of retinal cells decreased gradually with increasing LED color temperature. The new low-color-temperature LEDs caused less death and adverse effects on proliferation than the fluorescent white LEDs. After irradiation with high-color-temperature LEDs, the expression of Zonula Occludens-1 (ZO-1) was decreased and discontinuous in ARPE-19 cells; the stress protein hemeoxygenase-1 (HO-1) was upregulated in R28 cells; and glial fibrillary acidic protein (GFAP) and vimentin were upregulated in rMC-1 cells. We therefore conclude that the new white LEDs cause almost no damage to retinal cells and reduce the potential human health risks of chronic exposure to fluorescent white LEDs.


The Effect of Pressure on Organic Reactions in Fluids-a New Theoretical Perspective.
This Review brings a new perspective to the study of chemical reactions in compressed fluid media. We begin by reviewing the substantial insight gained from more than 50 years of experimental studies on organic reactions in solution under pressure. These led to a proper estimation of the critical roles of volume of activation (Δ≠ V) and reaction volume (ΔV) in understanding pressure effect on rates and equilibria of organic reactions. A recently developed computational method, the XP-PCM (extreme pressure polarizable continuum model) method, capable of carrying out quantum mechanical calculations of reaction pathways of molecules under pressure, is introduced. A case study of the Diels-Alder cycloaddition of cyclopentadiene with ethylene serves, in pedagogical detail, to describe the methodology. We then apply the XP-PCM method to a selection of other pericyclic reactions, including the parent Diels-Alder cycloaddition of butadiene with ethylene, electrocyclic ring-opening of cyclobutene, electrocyclic ring-closing of Z-hexatriene, the [1,5]-H shift in Z-pentadiene, and the Cope rearrangement. These serve as examples of some of the most common combinations of Δ≠ V and ΔV. Interesting phenomena such as a shift in a transition state along a reaction coordinate, a switch of rate-determining step, and the possible turning of a transition state into a stable minimum are revealed by the calculations. A reaction volume profile, the change in the volume of the reacting molecules as the reaction proceeds, emerges as being useful.


Immunological mechanisms of allergen-specific immunotherapy.
The studies on the mechanisms of specific immunotherapy (SIT) point out its targets that decide on the efficacy of SIT and hence might be used for its further improvement. Several mechanisms have been proposed to explain the beneficial effects of immunotherapy. The knowledge of the mechanisms underlying allergic diseases and curative treatment possibilities has experienced exciting advances over the last three decades. Studies in several clinical trials in allergen-SIT have demonstrated that the induction of a tolerant state against allergens in many ways represents a key step in the development of a healthy immune response against allergens. Several cellular and molecular mechanisms have been demonstrated: allergen-specific suppressive capacities of both inducible subsets of CD4(+) CD25(+) forkhead box P3(+) T-regulatory and IL-10-secreting type 1 T-regulatory cells increase in peripheral blood; suppression of eosinophils, mast cells, and basophils; Ab isotype change from IgE to IgG4. This review aims at the better understanding of the observed immunological changes associated with allergen SIT.


[Levels of sgRNA as a Major Factor Affecting CRISPRi Knockdown Efficiency in K562 Cells].
To determine how nuclease deactivated Cas9 (dCas9) or single-guide RNA (sgRNA) expression levels affect the knockdown efficiency of CRISPRi, we created K562 cell clones expressing KRAB-dCas9 protein either with the inducible Tet-on system or with the constitutive SFFV promotor. Single clones were selected by fluorescence-activated cell sorting (FACS) for further study. Six genes with various expression levels were targeted using lentiviral sgRNA from two libraries in four cell clones with various KRAB-dCas9 expression levels. The expression level of dCas9 protein/sgRNA levels and the knockdown efficiency were determined by flow cytometry. The cell clone with the highest KRAB-dCas9 expression level achieved effective CRISPRi knockdown. The data describing this clone were statistically different from that on other clones, indicating the strong KRAB-dCas9 expression might be a prerequisite for CRISPRi. By adopting different multiplicity of infection (MOI) in lentiviral transduction of this clone, we modified the expression level of sgRNA and found that the knockdown efficiency was neither affected by the target gene expression level nor correlated with KRAB-dCas9 levels, which remained relatively constant across all knockdown experiments (coefficient of variation = 2.2%). As an example, the following levels of the knockdowns: 74.72, 72.28 and 39.08% for mmadhc, rpia and znf148 genes, respectively, were achieved. These knockdown efficiencies correlated well with the respective sgRNA expression levels. Linear regression models built using this data indicate that the knockdown efficiency may be significantly affected by the levels of both KRAB-dCas9 and sgRNA. Notably, the sgRNA levels have greater impact, being a major factor affecting CRISPRi efficiency.


Immunogenicity of COVID-19 mRNA Vaccines in Pregnant and Lactating Women.
Pregnant women are at increased risk of morbidity and mortality from COVID-19 but have been excluded from the phase 3 COVID-19 vaccine trials. Data on vaccine safety and immunogenicity in these populations are therefore limited. To evaluate the immunogenicity of COVID-19 messenger RNA (mRNA) vaccines in pregnant and lactating women, including against emerging SARS-CoV-2 variants of concern. An exploratory, descriptive, prospective cohort study enrolled 103 women who received a COVID-19 vaccine from December 2020 through March 2021 and 28 women who had confirmed SARS-CoV-2 infection from April 2020 through March 2021 (the last follow-up date was March 26, 2021). This study enrolled 30 pregnant, 16 lactating, and 57 neither pregnant nor lactating women who received either the mRNA-1273 (Moderna) or BNT162b2 (Pfizer-BioNTech) COVID-19 vaccines and 22 pregnant and 6 nonpregnant unvaccinated women with SARS-CoV-2 infection. SARS-CoV-2 receptor binding domain binding, neutralizing, and functional nonneutralizing antibody responses from pregnant, lactating, and nonpregnant women were assessed following vaccination. Spike-specific T-cell responses were evaluated using IFN-γ enzyme-linked immunospot and multiparameter intracellular cytokine-staining assays. Humoral and cellular immune responses were determined against the original SARS-CoV-2 USA-WA1/2020 strain as well as against the B.1.1.7 and B.1.351 variants. This study enrolled 103 women aged 18 to 45 years (66% non-Hispanic White) who received a COVID-19 mRNA vaccine. After the second vaccine dose, fever was reported in 4 pregnant women (14%; SD, 6%), 7 lactating women (44%; SD, 12%), and 27 nonpregnant women (52%; SD, 7%). Binding, neutralizing, and functional nonneutralizing antibody responses as well as CD4 and CD8 T-cell responses were present in pregnant, lactating, and nonpregnant women following vaccination. Binding and neutralizing antibodies were also observed in infant cord blood and breast milk. Binding and neutralizing antibody titers against the SARS-CoV-2 B.1.1.7 and B.1.351 variants of concern were reduced, but T-cell responses were preserved against viral variants. In this exploratory analysis of a convenience sample, receipt of a COVID-19 mRNA vaccine was immunogenic in pregnant women, and vaccine-elicited antibodies were transported to infant cord blood and breast milk. Pregnant and nonpregnant women who were vaccinated developed cross-reactive antibody responses and T-cell responses against SARS-CoV-2 variants of concern.


A Comprehensive Tool for Usability Evaluation of Telehealth.
Telehealth has a high potential for delivering healthcare by overcoming the geographical distance, increased access to care, and efficiency. To obtain the potential benefits of telehealth, this system should be usable to the patients and healthcare providers. The aim of this study was to report a new comprehensive tool for assessing the usability of telehealth services and systems, covering different aspects of usability. We searched the keywords, such as telemedicine, telehealth, usability, satisfaction, questionnaires, and evaluation in Google, Scopus, PubMed, and Google Scholar from 1988 to 2019. All the studies that used the Telehealth services and computer& information technology evaluation tools were included. We identified 204 items and extracted 10 criteria that were classified into two categories. The first category is system usability, which includes Efficiency, Memorability, Learnability, Usefulness, and Satisfaction. Second category system reliability, included Interaction, Interface design, Saving, System capabilities, and Getting rid of Errors. This tool consider many aspects of telehealth service evaluation from the usability aspect. Further studies are warranted to evaluate other aspects of Telehealth services and measuring reliability so that researchers can have access to a comprehensive tool for assessing interventions in the field of Telemedicine and Telehealth.


[The role of arachidonic acid derivatives in generating membrane potential oscillations of the rat aorta endothelium].
Involvement of metabolites of arachidonic acid in electrical responses of rat aortic endothelium evoked by the agonist of aa1-adrenoreceptors, phenylephrine has been investigated. Our experiments suggest that phenylephrine evokes synthesis of thromboxane A2 in the rat aorta that is involved in the phenylephrine-evoked oscillations probably by increasing excitatory effect of the vasoconstrictor on vascular smooth muscle by the mechanism of positive feedback.


Phospholemman Ser69 phosphorylation contributes to sildenafil-induced cardioprotection against reperfusion injury.
The phosphodiesterase type-5 inhibitor sildenafil has powerful cardioprotective effects against ischemia-reperfusion injury. PKG-mediated signaling has been implicated in this protection, although the mechanism and the downstream targets of this kinase remain to be fully elucidated. In this study we assessed the role of phospholemman (PLM) phosphorylation, which activates the Na(+)/K(+)-ATPase, in cardioprotection afforded by sildenafil administered during reperfusion. Isolated perfused mouse hearts were optimally protected against infarction (indexed by tetrazolium staining) by 0.1 muM sildenafil treatment during the first 10 min of reperfusion. Extended sildenafil treatment (30, 60, or 120 min at reperfusion) did not alter the degree of protection provided. This protection was PKG dependent, since it was blocked by KT-5823. Western blot analysis using phosphospecific antibodies to PLM showed that sildenafil at reperfusion did not modulate PLM Ser63 or Ser68 phosphorylation but significantly increased Ser69 phosphorylation. The treatment of isolated rat ventricular myocytes with sildenafil or 8-bromo-cGMP (PKG agonist) enhanced PLM Ser69 phosphorylation, which was bisindolylmaleimide (PKC inhibitor) sensitive. Patch-clamp studies showed that sildenafil treatment also activated the Na(+)/K(+)-ATPase, which is anticipated in light of PLM Ser69 phosphorylation. Na(+)/K(+)-ATPase activation during reperfusion would attenuate Na(+) overload at this time, providing a molecular explanation of how sildenafil guards against injury at this time. Indeed, using flame photometry and rubidium uptake into isolated mouse hearts, we found that sildenafil enhanced Na(+)/K(+)-ATPase activity during reperfusion. In this study we provide a molecular explanation of how sildenafil guards against myocardial injury during postischemic reperfusion.


[Clinical and experimental study of the hemodynamic effect of gemiton].
The effect of hemiton (clofelin) on the minute circulation volume, general peripheral resistance to the blood flow, and the mechanisms of the regulation of the vascular neurogenous tonus and heart activity was studied in patients in the clinic and in experiments. It is shown that the hemodynamic effect of the agent depends on the initial parameters of hemodynamics. It is assumed that hemiton influences the capacity of systemic circulation. The expediency of combining hemiton with drugs which reduce the minute circulation volume is discussed.


Hysterectomy Uniquely Impacts Spatial Memory in a Rat Model: A Role for the Nonpregnant Uterus in Cognitive Processes.
Approximately one-third of women experience hysterectomy, or the surgical removal of the uterus, by 60 years of age, with most surgeries occurring prior to the onset of natural menopause. The ovaries are retained in about half of these surgeries, whereas for the other half hysterectomy occurs concurrently with oophorectomy. The dogma is that the nonpregnant uterus is dormant. There have been no preclinical assessments of surgical variations in menopause, including hysterectomy, with and without ovarian conservation, on potential endocrine and cognitive changes. We present a novel rat model of hysterectomy alongside sham, ovariectomy (Ovx), and Ovx-hysterectomy groups to assess effects of surgical menopause variations. Rats without ovaries learned the working memory domain of a complex cognitive task faster than did those with ovaries. Moreover, uterus removal alone had a unique detrimental impact on the ability to handle a high-demand working memory load. The addition of Ovx, that is, Ovx-hysterectomy, prevented this hysterectomy-induced memory deficit. Performance did not differ amongst groups in reference memory-only tasks, suggesting that the working memory domain is particularly sensitive to variations in surgical menopause. Following uterus removal, ovarian histology and estrous cycle monitoring demonstrated that ovaries continued to function, and serum assays indicated altered ovarian hormone and gonadotropin profiles by 2 months after surgery. These results underscore the critical need to further study the contribution of the uterus to the female phenotype, including effects of hysterectomy with and without ovarian conservation, on the trajectory of brain and endocrine aging to decipher the impact of common variations in gynecological surgery in women. Moreover, findings demonstrate that the nonpregnant uterus is not dormant, and indicate that there is an ovarian-uterus-brain system that becomes interrupted when the reproductive tract has been disrupted, leading to alterations in brain functioning.


Stage IV and age over 45 years are the only prognostic factors of the International Prognostic Score for the outcome of advanced Hodgkin lymphoma in the Spanish Hodgkin Lymphoma Study Group series.
The International Prognostic Score (IPS) is the most widely used system to date for identifying risk groups for the outcome of patients with advanced Hodgkin lymphoma, although important limitations have been recognized. We analyzed the value of the IPS in a series of 311 patients with advanced classical Hodgkin lymphoma (cHL) (Ann Arbor stage III, IV or stage II with B symptoms and/or bulky masses) treated with first-line chemotherapy including adriamycin (adriamycin, bleomycin, vinblastine, dacarbazine [ABVD] or equivalent variants). In univariate and multivariate analyses, stage IV disease and age ≥ 45 years were the only factors with independent predictive significance for overall survival (OS) (p = 0.002 and p < 0.001, respectively). Stage IV was still significant for freedom from progression (FFP) (p = 0.001) and age ≥ 45 years was borderline significant (p = 0.058). IPS separates prognostic groups, as in the original publication, but this is mainly due to the high statistical significance of stage IV and age ≥ 45 years. Moreover, the combination of these two factors enables a simpler system to be constructed that separates groups with different FFP and OS. In conclusion, in our series, stage IV and age ≥ 45 years are the key prognostic factors for the outcome of advanced cHL.


Attachment, mentalisation and expressed emotion in carers of people with long-term mental health difficulties.
Expressed emotion (EE) is a global index of familial emotional climate, which is comprised of emotional over-involvement (EOI) and critical comments (CC)/hostility. Although EE is an established predictor of negative outcomes for both people with long-term mental health difficulties and their family carers, its psychological underpinnings remain relatively poorly understood. This paper examined associations between attachment, mentalisation ability and aspects of EE. Carers of people with long-term mental health difficulties (n = 106) completed measures of adult attachment (the Experiences in Close Relationships-Short Form questionnaire), mentalisation (the Reading the Mind in the Eyes Test and the Emotional Self-Efficacy Scale) and EE (the Family Questionnaire). Data were analysed using hierarchical multiple regression. Attachment avoidance and facets of mentalisation were directly and uniquely positively associated with CC/hostility, with attachment avoidance and other-directed emotional self-efficacy (one facet of mentalisation) each significantly predicting CC/hostility scores after controlling for the effects of EOI and demographic variables. However, no associations were observed between EOI, attachment anxiety and mentalisation. Furthermore, no indirect effects from attachment to EE via mentalisation was found. Although it would be premature to propose firm clinical implications based on these findings, data indicate that it may be beneficial for clinicians to consider attachment and mentalisation in their conceptualisation of carers' criticism and hostility. However, further research is needed to clarify the magnitude of these associations and their direction of effect before firm conclusions can be drawn.


Spontaneous intracranial hypotension: Two cases including one treated with epidural blood patch.
Spontaneous intracranial hypotension (SIH) is characterized by orthostatic headache (OH), low cerebrospinal fluid (CSF) pressure, and diffuse pachymeningeal gadolinium enhancement (DPME). We present here the case studies of two patients. One patient demonstrated a CSF leak in the mid-thoracic region, and recovered completely with conservative treatment. The other patient in whom leak could not be demonstrated, developed dementia, rapidly worsening encephalopathy, and became comatose, necessitating urgent epidural blood patch (EBP) with 25 cc of autologous blood, after which immediate and complete symptomatic relief was obtained. A second EBP was required a few days later and also provided complete and sustained clinical benefit, without subsequent recurrence. Both patients had OH and showed bilateral subdural fluid collections, DPME and "sagging" of brain on MRI. A high index of suspicion, recognizing the orthostatic nature of headache, and typical findings on contrast enhanced MRI should point to the diagnosis of SIH. EBP can be effective treatment in patients unresponsive to conservative measures.


Unravelling migrants' health paradoxes: a transdisciplinary research agenda.
The Social Determinants of Health literature has consistently found that a higher socioeconomic status is associated with better health outcomes even after adjusting for traditional risk factors. However, research findings in the field of Migrants' Health suggest that the socioeconomic/health gradient does not always behave as expected for migrants and their descendants. The mismatch of findings in these two long-standing parallel research traditions is exemplified by frequent reports of paradoxical findings in the scientific literature: the healthy migrant paradox, the ethnic density paradox and the diminishing returns paradox. This paper outlines a transdisciplinary research agenda to elucidate the social processes that underpin these disconcerting findings and calls for a shift from a pathogenic deficit model that sees migrants as a burden to their reconceptualisation as actively engaged citizens in search of solutions. Amidst a severe refugee crisis, fears of terrorist attacks and political capitalisation of these tragedies to foster antimigrant sentiments, this is urgently needed.


The diagnosis of beta-thalassemia trait: a review.
Diagnostic and screening methods for beta-thalassemia trait are reviewed in the order of their development, including identification by homozygous offspring, erythrocyte morphology, osmotic fragility, hemoglobin composition, globin synthetic rates, and red blood cell indices.


Thioaptamer targeted discoidal microparticles increase self immunity and reduce Mycobacterium tuberculosis burden in mice.
Worldwide, tuberculosis (TB) remains one of the most prevalent infectious diseases causing morbidity and death in >1.5 million patients annually. Mycobacterium tuberculosis (Mtb), the etiologic agent of TB, usually resides in the alveolar macrophages. Current tuberculosis treatment methods require more than six months, and low compliance often leads to therapeutic failure and multidrug resistant strain development. Critical to improving TB-therapy is shortening treatment duration and increasing therapeutic efficacy. In this study, we sought to determine if lung hemodynamics and pathological changes in Mtb infected cells can be used for the selective targeting of microparticles to infected tissue(s). Thioaptamers (TA) with CD44 (CD44TA) targeting moiety were conjugated to discoidal silicon mesoporous microparticles (SMP) to enhance accumulation of these agents/carriers in the infected macrophages in the lungs. In vitro, CD44TA-SMP accumulated in macrophages infected with mycobacteria efficiently killing the infected cells and decreasing survival of mycobacteria. In vivo, increased accumulations of CD44TA-SMP were recorded in the lung of M. tuberculosis infected mice as compared to controls. TA-targeted carriers significantly diminished bacterial load in the lungs and caused recruitment of T lymphocytes. Proposed mechanism of action of the designed vector accounts for a combination of increased uptake of particles that leads to infected macrophage death, as well as, activation of cellular immunity by the TA, causing increased T-cell accumulation in the treated lungs. Based on our data with CD44TA-SMP, we anticipate that this drug carrier can open new avenues in TB management.


[The importance of echography in the diagnosis of precancer and cancer of the endometrium].
Ten-year experience with ultrasonography was gained at an outpatient center. Within this period, 27000 examinations were performed in 11996 females. The credibility of the method in diagnosing endometrial pathology was 85.8%. Cancer morbidity for the last 5 years was compared in the ultrasonography and no ultrasonography groups. Endometrial cancer morbidity was 6.2 times higher in the latter group. Large-scale application of the procedure followed by adequate treatment assures significantly lower incidence of said pathology.


Determinants of Ocular Pain Severity in Patients With Dry Eye Disease.
To quantify the severity of ocular pain in patients with dry eye disease (DED) and evaluate factors associated with pain severity. Cross-sectional study. Eighty-four patients with DED were asked to score their severity level of ocular pain using a 10-point scale, with 10 indicating the most severe pain. All patients also had a comprehensive ophthalmic assessment including a detailed history, Ocular Surface Disease Index (OSDI) questionnaire, and ocular surface examination. Regression analysis was used to determine the factors associated with ocular pain severity. The mean OSDI score was 45.6 ± 23.1. At least some degree of ocular pain (score >1) was reported by 88.1% of patients, including mild pain (scores 2-4) in 46.4%, moderate pain (scores 5-7) in 34.5%, and severe pain (scores 8-10) in 7.1% of patients. Ocular pain levels significantly correlated with the OSDI score (rs = 0.49, P < .001). Regression analysis showed that the severity of ocular pain had a significant association with use of antidepressant medications (P = .045) but not with tear breakup time, corneal fluorescein staining, or ocular medications used by patients. In patients without pain, a significant correlation was seen between OSDI and corneal fluorescein staining scores (rs = 0.67, P = .01). However, such a correlation was not observed in those with ocular pain. A majority of patients with DED report some degree of ocular pain, which correlates only moderately with the OSDI score. Severity of ocular pain correlates with nonocular comorbidities such as use of antidepressant medications rather than with clinical signs of DED.


Extracellular guanosine and guanine nucleotides decrease viability of human breast cancer SKBR-3 cells.
Though the physiological effects of adenosine and adenine nucleotides on purinergic receptors in cancer cells have been well studied, the influence of extracellular guanosine and guanine nucleotides on breast cancer cells remains unclear. Here, we show that extracellular guanosine and guanine nucleotides decrease the viability and proliferation of human breast cancer SKBR-3 cells. Treatment with guanosine or guanine nucleotides increased mitochondrial production of reactive oxygen species (ROS), and modified the cell cycle. Guanosine-induced cell death was suppressed by treatment with adenosine or the equilibrium nucleoside transporter (ENT) 1/2 inhibitor dipyridamole, but was not affected by adenosine receptor agonists or antagonists. These results suggest that guanosine inhibits adenosine uptake through ENT1/2, but does not antagonize adenosine receptors. In contrast, GTP-induced cell death was suppressed not only by adenosine and dipyridamole, but also by the A1 receptor agonist CCPA, suggesting that GTP-induced cell death is mediated in part by an antagonistic effect on adenosine A1 receptor. Thus, both guanosine and GTP induce apoptosis of breast cancer cells, but via at least partially different mechanisms.


Overview of obesity in Mainland China.
This overview describes the study of obesity issues over the last 8 years in Mainland China. A disease risk-based body mass index classification for Chinese adults was developed and verified by the results of the 2002 China Nationwide Nutrition and Health Survey. Age and gender-specific BMI cut-offs for Chinese children and adolescents aged 7-18 years were also developed. As the first step in the obesity control effort, the public health implications of the prevalence of overweight and obesity were set out and publicized by the government of China. The overweight prevalence in 2002 was 22.8% and for obesity 7.1%, these having increased by 40.7% and 97.2%, respectively, since 1992. In children, dramatic increases in overweight/obesity have occurred with a two- to threefold increase in Beijing and Shanghai between 1985 and 1995; the prevalence of overweight plus obesity in 7- to 12-year-old boys approached 29% in 2000 and in girls 15-17%. The rapid development of overweight/obesity in China will undoubtedly push up the prevalence of chronic disease. The relative risk for adult hypertension and diabetes in the overweight and obese populations was 2-5 in 2002. In adolescents, the relative risk of hypertension in the overweight was 3.3 and in the obese 3.9, with a clustering of cardiovascular risk factors in the obese Chinese children of whom 90.5% had evidence of abdominal obesity. Even the overweight group had 15.8% with abdominal obesity. Based on the population attributable risks of overweight and obesity in 2002 and data on the direct medical costs of hypertension, diabetes, stroke and coronary heart disease, 25% of these comorbidity costs could be attributed to overweight and obesity. As the improvement in linear growth of young children is usually slower than their improvements in weight during the economic development of China, the long-term adult implications of having a problem of 'Stunted obesity' rather than simple obesity in young children are raised. Analysis of the associations between adult obesity with the individuals' fetal and early nutrition highlights the need for further investigation.


Development of a brief diabetes distress screening instrument.
Previous research has documented that diabetes distress, defined as patient concerns about disease management, support, emotional burden, and access to care, is an important condition distinct from depression. We wanted to develop a brief diabetes distress screen instrument for use in clinical settings. We assessed 496 community-based patients with type 2 diabetes on the previously validated, 17-item Diabetes Distress Scale (DDS17) and 6 biobehavioral measures: glycated hemoglobin (HbA(1c)); non-high-density-lipoprotein (non-HDL) cholesterol; kilocalories, percentage of calories from fat, and number of fruit and vegetable servings consumed per day; and physical activity as measured by the International Physical Activity Questionnaire. An average item score of > or =3 (moderate distress) discriminated high- from low-distressed subgroups. The 4 DDS17 items with the highest correlations with the DDS17 total (r = .56-.61) were selected. Composites, comprised of 2, 3, and 4 of these items (DDS2, DDS3, DDS4), yielded higher correlations (r=.69-.71). The sensitivity and specificity of the composites were .95 and .85, .93 and .87, and .97 and .86, respectively. The DDS3 had a lower sensitivity and higher percentages of false-negative and false-positive results. All 3 composites significantly discriminated subgroups on HbA(1c), non-HDL cholesterol, and kilocalories consumed per day; none discriminated subgroups on fruit and vegetable servings consumed per day; and only the DDS3 yielded significant results on the International Physical Activity Questionnaire. Because of its psychometric properties and brevity, the DDS2 was selected as a screening instrument. The DDS2 is a 2-item diabetes distress screening instrument asking respondents to rate on a 6-point scale the degree to which the following items caused distress: (1) feeling overwhelmed by the demands of living with diabetes, and (2) feeling that I am often failing with my diabetes regimen. The DDS17 can be administered to those who have positive findings on the DDS2 to define the content of distress and to direct intervention.


Effect of the oral absorption of benzenesulfonanilide-type cyclooxygenase-1 inhibitors on analgesic action and gastric ulcer formation.
A benzensulfonanilide-type cyclooxygenase-1 (COX-1)-selective inhibitor, ZXX2-77: 4-amino-4'-chloro-N-methylbenzenesulfonanilide (4a), has been reported as a novel analgesic that does not cause gastric damage. This compound has a weak analgesic effect but has potent in vitro COX-1 inhibitory activity. Since the reason for the weak analgesic effect in vivo was thought to be the low rate of oral absorption, the blood concentration of ZXX2-77 (4a) was measured in rats. It was found that the C(max) value (1.2 microM) of ZXX2-77 (4a) at a dose of 30 mg/kg did not reach the COX-1 IC(50) value (3.2 microM). On the other hand, ZXX2-79 (4b) (SO(2)NH derivative of ZXX2-77 (4a); 4-amino-4'-chlorobenzenesulfonanilide), which shows less potent COX inhibitory activities (COX-1 IC(50) = 12 microM, COX-2 IC(50) = 150 microM) than those of ZXX2-77 (4a) in vitro, was found to be more absorbable (C(max) = 16 microM at a dose of 30 mg/kg in rats) than ZXX2-77 (4a). Furthermore, ZXX2-79 (4b) not only showed a potent analgesic effect in a formalin test but also caused little gastric damage. These findings indicate that demethylated sulfonamide compounds are more easily absorbed than are N-methylated sulfonamide compounds and suggest that COX-1-selective inhibitors will be useful as analgesics that do not cause gastric damage.


A novel function of the MA-3 domains in transformation and translation suppressor Pdcd4 is essential for its binding to eukaryotic translation initiation factor 4A.
An alpha-helical MA-3 domain appears in several translation initiation factors, including human eukaryotic translation initiation factor 4G (eIF4G) and DAP-5/NAT1/p97, as well as in the tumor suppressor Pdcd4. The function of the MA-3 domain is, however, unknown. C-terminal eIF4G (eIG4Gc) contains an MA-3 domain that is located within the eIF4A-binding region, suggesting a role for eIF4A binding. Interestingly, C-terminal DAP-5/NAT1/p97 contains an MA-3 domain, but it does not bind to eIF4A. Mutation of amino acid residues conserved between Pdcd4 and eIF4Gc but not in DAP-5/NAT1/p97 to the amino acid residues found in the DAP-5/NAT1/p97 indicates that some of these amino acid residues within the MA-3 domain are critical for eIF4A-binding activity. Six Pdcd4 mutants (Pdcd4(E249K), Pdcd4(D253A), Pdcd4(D414K), Pdcd4(D418A), Pdcd4(E249K,D414K), and Pdcd4(D253A,D418A)) lost >90% eIF4A-binding activity. Mutation of the corresponding amino acid residues in the eIF4Gc also produced similar results, as seen for Pdcd4. These results demonstrate that the MA-3 domain is important for eIF4A binding and explain the ability of Pdcd4 or eIF4Gc but not DAP-5/NAT1/p97 to bind to eIF4A. Competition experiments indicate that Pdcd4 prevents ca. 60 to 70% of eIF4A binding to eIF4Gc at a Pdcd4/eIF4A ratio of 1:1, but mutants Pdcd4(D253A) and Pdcd4(D253A,D418A) do not. Translation of stem-loop structured mRNA is susceptible to inhibition by wild-type Pdcd4 but not by Pdcd4(D253A), Pdcd4(D418A), or Pdcd4(D235A,D418A). Together, these results indicate that not only binding to eIF4A but also prevention of eIF4A binding to the MA-3 domain of eIF4Gc contributes to the mechanism by which Pdcd4 inhibits translation.


Characterization of sodium-dependent and sodium-independent nucleoside transport systems in rabbit brush-border and basolateral plasma-membrane vesicles from the renal outer cortex.
The transport of uridine into rabbit renal outer-cortical brush-border and basolateral membrane vesicles was compared at 22 degrees C. Uridine was taken up into an osmotically active space in the absence of metabolism for both types of membrane vesicles. Uridine influx by brush-border membrane vesicles was stimulated by Na+, and in the presence of inwardly directed gradients of Na+ a transient overshoot phenomenon was observed, indicating active transport. Kinetic analysis of the saturable Na+-dependent component of uridine flux indicated that it was consistent with Michaelis-Menten kinetics (Km 12 +/- 3 microM, Vmax. 3.9 +/- 0.9 pmol/s per mg of protein). The sodium:uridine coupling stoichiometry was found to be consistent with 1:1 and involved the net transfer of positive charge. In contrast, uridine influx by basolateral membrane vesicles was not dependent on the cation present and was inhibited by nitrobenzylthioinosine (NBMPR). NBMPR-sensitive uridine transport was saturable (Km 137 +/- 20 microM, Vmax. 5.2 +/- 0.6 pmol/s per mg of protein). Inhibition of uridine flux by NBMPR was associated with high-affinity binding of NBMPR to the basolateral membrane (Kd 0.74 +/- 0.46 nM). Binding of NBMPR to these sites was competitively blocked by adenosine and uridine. These results indicate that uridine crosses the brush-border surface of rabbit proximal renal tubule cells by Na+-dependent pathways, but permeates the basolateral surface by NBMPR-sensitive facilitated-diffusion carriers.


Origin of the metazoan phyla: molecular clocks confirm paleontological estimates.
The time of origin of the animal phyla is controversial. Abundant fossils from the major animal phyla are found in the Cambrian, starting 544 million years ago. Many paleontologists hold that these phyla originated in the late Neoproterozoic, during the 160 million years preceding the Cambrian fossil explosion. We have analyzed 18 protein-coding gene loci and estimated that protostomes (arthropods, annelids, and mollusks) diverged from deuterostomes (echinoderms and chordates) about 670 million years ago, and chordates from echinoderms about 600 million years ago. Both estimates are consistent with paleontological estimates. A published analysis of seven gene loci that concludes that the corresponding divergence times are 1,200 and 1,000 million years ago is shown to be flawed because it extrapolates from slow-evolving vertebrate rates to faster-evolving invertebrate rates, as well as in other ways.


Targeting antioxidant and antithrombotic biotherapeutics to endothelium.
The endothelium is one of the key targets for pharmacological interventions in oxidative stress and thrombosis, two conditions that are notoriously difficult to treat due to limited efficacy and precision of action of current drugs. Design of molecular and nano-devices that deliver potent antioxidant and antithrombotic therapeutic enzymes to the endothelium holds promise to improve the potency, localization, timing, specificity, safety, and mechanistic precision of these interventions. In particular, cell adhesion molecules expressed on the surface of resting and pathologically altered endothelial cells can be used for drug delivery to the endothelial surface (preferable for thrombolytics) and into intracellular compartments (preferable for antioxidants). Drug delivery platforms including protein conjugates, recombinant fusion constructs, and stealth polymer carriers designed to target these drugs to endothelium are reviewed in this article.


Effects of Citrulline Supplementation on Body Weight in Patients With Hepatobiliary Pancreatic Surgery.
Many health benefits have been proposed for citrulline, but they lack evidence based on human research. This study was to evaluate whether an oral citrulline supplement affects body weight changes and laboratory values in patients with hepatobiliary and pancreatic surgery. Patients who underwent hepatobiliary and pancreatic surgery during January to June 2015 were screened for analysis. Patients using citrulline during hospital stay and at discharge were classified as the citrulline group, whereas those without any records of citrulline were designated as the control group. The primary efficacy outcome was the change in body weight at discharge and at first outpatient visit. Other outcomes included change in laboratory values. A total of 138 patients were included in analysis. Citrulline group and control group did not differ with respect to baseline characteristics except for white blood cell count. Percent in change of body weight and body mass index from discharge to first outpatient visit was significantly different between the 2 groups, showing less weight loss in citrulline group than in controls (-0.8 ± 2.7% vs -2.5 ± 3.8%, P < 0.05). Especially in men, citrulline use significantly affected weight loss from the multivariate analysis (P < 0.05); percent change in weight in citrulline group was predicted to increase by 2.1 units. During hospital stay, significant differences between the 2 groups were found in changes of cholesterol and protein levels. Citrulline supplement reduced weight loss in surgical patients during recovery.


Effects of Hepatitis B Surface Antigen on Virus-Specific and Global T Cells in Patients With Chronic Hepatitis B Virus infection.
Chronic hepatitis B virus (HBV) infection is characterized by the presence of defective viral envelope proteins (hepatitis B surface antigen [HBsAg]) and the duration of infection-most patients acquire the infection at birth or during the first years of life. We investigated the effects of these factors on patients' lymphocyte and HBV-specific T-cell populations. We collected blood samples and clinical data from 243 patients with HBV infection (3-75 years old) in the United Kingdom and China. We measured levels of HBV DNA, HBsAg, hepatitis B e antigen, and alanine aminotransferase; analyzed HBV genotypes; and isolated peripheral blood mononuclear cells (PBMCs). In PBMCs from 48 patients with varying levels of serum HBsAg, we measured 40 markers on nature killer and T cells by mass cytometry. PBMCs from 189 patients with chronic infection and 38 patients with resolved infections were incubated with HBV peptide libraries, and HBV-specific T cells were identified by interferon gamma enzyme-linked immune absorbent spot (ELISpot) assays or flow cytometry. We used multivariate linear regression and performed variable selection using the Akaike information criterion to identify covariates associated with HBV-specific responses of T cells. Although T- and natural killer cell phenotypes and functions did not change with level of serum HBsAg, numbers of HBs-specific T cells correlated with serum levels of HBsAg (r = 0.3367; P < .00001). After we performed the variable selection, the multivariate linear regression model identified patient age as the only factor significantly associated with numbers of HBs-specific T cells (P = .000115). In patients younger than 30 years, HBs-specific T cells constituted 28.26% of the total HBV-specific T cells; this value decreased to 7.14% in patients older than 30 years. In an analysis of immune cells from patients with chronic HBV infection, we found that the duration of HBsAg exposure, rather than the quantity of HBsAg, was associated with the level of anti-HBV immune response. Although the presence of HBs-specific T cells might not be required for the clearance of HBV infection in all patients, strategies to restore anti-HBV immune responses should be considered in patients younger than 30 years.


Moving Together While Staying Apart: Practical Recommendations for 24-Hour Home-Based Movement Behaviours for Those With Cardiovascular Disease.
The novel coronavirus disease 2019 is a global public health crisis that disproportionately affects those with preexisting conditions. Cardiovascular disease (CVD) is the leading cause of death worldwide and many key CVD risk factors are modifiable (e.g., physical inactivity, sedentary behaviour, obesity). To limit the spread of coronavirus 2019, most governments have implemented restrictions and recommended staying at home, reducing social contact to a select and exclusive few, and limiting large gatherings. Such public health constraints may have unintended, negative health consequences on 24-hour movement behaviours. The primary purpose of this review is to provide practical at-home recommendations for sedentary time, sleep, and physical activity in those living with CVD. Adults with CVD will benefit from practical recommendations to reduce sedentary time, increase purposeful exercise, and maintain optimal sleep patterns while being at home and adhering to public health restrictions. Our recommendations include the following: (i) self-monitoring sitting time; (ii) engaging in 2-3 days per week of purposeful exercise for those with low exercise capacity and > 3 days per week for those with moderate-to-high exercise capacity; (iii) self-monitoring exercise intensity through the use of scales or wearable devices; (iv) maintaining a regular sleep schedule; and (v) moving daily. Clinicians should be aware that clear communication of the importance of limiting prolonged sedentary time, engaging in regular physical activity and exercise, and ensuring optimal sleep in association with the provision of clear, comprehensible, and practical advice is fundamental to ensuring that those living with CVD respond optimally to the challenges posed by the pandemic.


Importance of metal hydration on the selectivity of Mg2+ versus Ca2+ in magnesium ion channels.
Magnesium ion channels and transporters regulate the cellular concentrations of Mg(2+), which must be tightly controlled as imbalances have been associated with diseases such as osteoporosis, diabetes, and high blood pressure in humans. The channels and transporters allow the "native" Mg(2+) to be transported against a high background concentration of its major competitor, Ca(2+). Their selectivity filters (the narrowest part of the open pore) control metal ion selectivity. As the structures of Mg(2+) channels in an open conformation with bound Mg(2+) have not yet been solved, the key determinants of Mg(2+)/Ca(2+) selectivity in Mg(2+) ion channels remain elusive. Here, using density functional theory combined with continuum dielectric methods, we evaluated how the competition between Mg(2+) and Ca(2+) in model selectivity filters depends on the degree of metal hydration, which correlates with the pore size/rigidity as well as the composition and solvent accessibility of the selectivity filter. The key determinant of the selectivity for Mg(2+) over Ca(2+) in the Mg(2+) channel selectivity filter is a pore that is sufficiently large to accommodate hexahydrated Mg ions. In such wide pores, the hexahydrated metal ions interact indirectly with the protein ligands, hence metal desolvation and ligand-ligand steric repulsion become less important than Mg(2+)-water-protein interactions. These wide pores are Mg(2+)-selective because compared to Ca(2+) or Na(+) and K(+) monocations, Mg(2+) better polarizes the bound water molecules resulting in stronger Mg(2+)-water-protein interactions. Although both tetrameric and pentameric filters with pores that can accommodate hexahydrated metal ions could select Mg(2+) over Ca(2+), a bilayered pentameric filter lined with a ring of amides and a ring of carboxylates seems to best discriminate the "native" Mg(2+) from its key rival, Ca(2+). Our results are consistent with available experimental data and help to elucidate the selectivity filters in the Mg(2+)-selective TRPM6 and CorA channels.


Ethylene Glycol Poisoning Should Not Contraindicate Liver Donation.
As the number of patients waiting to receive transplants increases, there is a need to explore all possible donation opportunities. In this case report, we describe the transplantation of a liver from a donor who died after ethylene glycol poisoning into a woman with alcoholic liver disease with cirrhosis and associated ascites. Donor management, including ethanol, fomepizol and haemodialysis, hastened clearance of ethylene glycol from the circulation, and after liver transplantation, the recipient exhibited no adverse effects suggestive of ethylene glycol toxicity, although recipient hepatic artery dissection and thrombosis necessitated retransplantation. Our experience suggests that donor death due to ethylene glycol intoxication should not contraindicate liver transplantation, particularly after appropriate donor management.


Small but not isolated: a population genetic survey of the tropical tree Cariniana estrellensis (Lecythidaceae) in a highly fragmented habitat.
Here, we explore the mating pattern and genetic structure of a tropical tree species, Cariniana estrellensis, in a small population in which progeny arrays (n=399), all adults (n=28) and all seedlings (n=39) were genotyped at nine highly informative microsatellite loci. From progeny arrays we were able to identify the source tree for at least 78% of pollination events. The gene immigration rates, mainly attributable to pollen, were high, varying from 23.5 to 53%. Although gene dispersal over long distance was observed, the effective gene dispersal distances within the small population were relatively short, with mean pollination distances varying from 69.9 to 146.9 m, and seed dispersal distances occurring up to a mean of 119.6 m. Mating system analyses showed that C. estrellensis is an allogamous species (tm=0.999), with both biparental inbreeding (tm-ts=-0.016) and selfing rates (s=0.001) that are not significantly different from zero. Even though the population is small, the presence of private alleles in both seedlings and progeny arrays and the elevated rates of gene immigration indicate that the C. estrellensis population is not genetically isolated. However, genetic diversity expressed by allelic richness was significantly lower in postfragmentation life stages. Although there was a loss of genetic diversity, indicating susceptibility of C. estrellensis to habitat fragmentation, no evidence of inbreeding or spatial genetic structure was observed across generations. Overall, C. estrellensis showed some resilience to negative genetic effects of habitat fragmentation, but conservation strategies are needed to preserve the remaining genetic diversity of this population.


Beyond guanine quartets: cation-induced formation of homogenous and chimeric DNA tetraplexes incorporating iso-guanine and guanine.
iso-Guanine (iso-G) is the purine component of an isomeric Watson-Crick base pair that may have existed prebiotically. By comparing the abiotic molecular recognition properties of iso-G and its complement, iso-cytosine (iso-C), with those of genomic nucleotide bases, it may be possible to explain the exclusion of the iso-G-iso-C base pair from modern genomes. Whether a nucleobase forms quartets may have a key role in determining its functionality. Biotically, nucleic acid tetraplexes have been implicated in cellular functions; prebiotically, tetraplexes would probably interfere with replication. Recently, in vitro selection has yielded receptors and catalysts that incorporate G quartets. The versatility of these structures could be enhanced by expanding the range of bases that can form the quartet motif. Native polyacrylamide gel electrophoresis of oligonucleotides bearing runs of iso-G provides strong support for tetraplex formation via cation-promoted DNA strand association. In particular, when strands of different lengths bearing the same iso-G tetrad recognition element were combined, five bands were observed after electrophoresis, corresponding to all possible heterotetraplexes with parallel strand alignment. An analogous experiment with a mixture of strands bearing iso-G or G tetrad recognition domains supports the existence of mixed iso-G/G tetraplexes with antiparallel strand alignment at chimeric junctions. iso-G tetraplex and quartet structure has also been probed by a photo-crosslinking experiment, ultra-violet spectroscopy and theoretical calculations. As iso-G and G both have a marked tendency to form tetraplexes, their tandem inclusion in genetic material may be problematic, leading to double-stranded DNA half composed of bases that have a tendency to auto-associate. The resulting density of 'selfish' bases could undermine Watson-Crick pair formation, especially in a prebiotic context devoid of enzymes. Nevertheless, the ability of iso-G to form mixed quartets with G may provide a basis for altering the properties of tetraplexes in the domain of artificial receptors or catalysts from in vitro selections.


Better defining the optimal management of penile urethral strictures: A retrospective comparison of single-stage vs. two-stage urethroplasty.
We aimed to compare single-stage and two-stage urethroplasty techniques for the treatment of penile urethral strictures. We performed a retrospective review of all penile urethroplasties performed at a single centre between 2003 and 2017. The primary outcome was urethral patency, defined as the ability to easily pass a 16 Fr flexible cystoscope at six and 18 months of followup, and development of 90-day complications. Overall, 101 single-stage procedures (48 buccal mucosal graft [BMG] and 53 penile fasciocutaneous flap [PFF]) and 53 two-stage procedures were performed. There was no difference in median stricture length between groups (p=0.25). Cox regression analysis did not identify stricture etiology, length, age, obesity, prior reconstruction, or urethroplasty technique to be associated with failure. Log-rank testing did not demonstrate a difference in success rates between surgical techniques (91% [48/53] PFF vs. 83% [40/48] BMG vs. 87% two-stage [46/53]). Thirty-nine percent (60/154) of patients experienced a complication (51% [27/53] PFF vs. 29% [14/48] BMG vs. 36% [19/53] two-stage). Multivariate analysis found urethroplasty technique to be the only factor associated with development of complication (p=0.02); odds ratio relative to BMG was 3.1 (p=0.009) for PFF and 1.4 (p=0.43) for two-stage. There appears to be little difference in success between penile urethroplasty techniques. The shift in technique to a single-stage BMG, when appropriate, appears to be founded on the basis of fewer operations for the patient, relative to a two-stage repair, and a lower complication profile, relative to single-stage PFF, without compromising success rates.


Intracellular pathways of insulin transport across vascular endothelial cells.
Processing and transport of hormones across vascular endothelial cells may modulate hormone action at subendothelial tissue sites. Insulin was transported across cultured rat capillary and bovine aortic endothelial cells, after a delay of 5-10 min, at a constant rate for 60 min at 37 degrees C. 125I-labeled insulin transport was inhibited by 88 +/- 11% (SE, n = 4) and 75 +/- 18% (SE, n = 4) in the presence of anti-insulin receptor antibody and unlabeled insulin (at 10(-7) M), respectively. Reverse phase high-performance liquid chromatography showed 88% of the 125I-insulin transported over 60 min was indistinguishable from the 125I-insulin added to the cells at 4 degrees C. In aortic endothelial cells preincubated with 2.3 x 10(-9) M of insulin for 24 h, insulin receptor binding was downregulated by 67%, and 125I-insulin transport was decreased by 52 +/- 11%. The proton ionophore monensin (0.05 mM) increased the internalized insulin in bovine aortic endothelial cells by 78%, with a corresponding decrease in 125I-insulin released by 76 +/- 2% (SE, n = 4). 125I-insulin transport across the aortic endothelial cell monolayer was similarly decreased (54 +/- 12%, SE, n = 4) by monensin. In contrast, the lysosomal protease inhibitor leupeptin had no effect. Degradation and transport were similarly dissociated by low temperature. At 15 degrees C, no significant insulin degradation was detected, whereas 125I-insulin release from the cells continued at 30 +/- 3% of the rate at 37 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluating the Need to Address Digital Literacy Among Hospitalized Patients: Cross-Sectional Observational Study.
Technology is a potentially powerful tool to assist patients with transitions of care during and after hospitalization. Patients with low health literacy who are predisposed to poor health outcomes are particularly poised to benefit from such interventions. However, this population may lack the ability to effectively engage with technology. Although prior research studied the role of health literacy in technology access/use among outpatients, hospitalized patient populations have not been investigated in this context. Further, with the rapid uptake of technology, access may no longer be pertinent, and differences in technological capabilities may drive the current digital divide. Thus, characterizing the digital literacy of hospitalized patients across health literacy levels is paramount. We sought to determine the relationship between health literacy level and technological access, use, and capability among hospitalized patients. Adult inpatients completed a technology survey that asked about technology access/use and online capabilities as part of an ongoing quality of care study. Participants' health literacy level was assessed utilizing the 3-question Brief Health Literacy Screen. Descriptive statistics, bivariate chi-squared analyses, and multivariate logistic regression analyses (adjusting for age, race, gender, and education level) were performed. Using Bonferroni correction for the 18 tests, the threshold P value for significance was <.003. Among 502 enrolled participants, the mean age was 51 years, 71.3% (358/502) were African American, half (265/502, 52.8%) were female, and half (253/502, 50.4%) had at least some college education. Over one-third (191/502, 38.0%) of participants had low health literacy. The majority of participants owned devices (owned a smartphone: 116/173, 67.1% low health literacy versus 235/300, 78.3% adequate health literacy, P=.007) and had used the Internet previously (143/189, 75.7% low health literacy versus 281/309, 90.9% adequate health literacy, P<.001). Participants with low health literacy were more likely to report needing help performing online tasks (133/189, 70.4% low health literacy versus 135/303, 44.6% adequate health literacy, P<.001). In the multivariate analysis, when adjusting for age, race, gender, and education level, we found that low health literacy was not significantly associated with a lower likelihood of owning smartphones (OR: 0.8, 95% CI 0.5-1.4; P=.52) or using the internet ever (OR: 0.5, 95% CI 0.2-0.9; P=.02). However, low health literacy remained significantly associated with a higher likelihood of needing help performing any online task (OR: 2.2, 95% CI 1.3-3.6; P=.002). The majority of participants with low health literacy had access to technological devices and had used the internet previously, but they were unable to perform online tasks without assistance. The barriers patients face in using online health information and other health information technology may be more related to online capabilities rather than to technology access. When designing and implementing technological tools for hospitalized patients, it is important to ensure that patients across digital literacy levels can both understand and use them.


Salmon blood plasma: effective inhibitor of protease-laden Pacific whiting surimi and salmon mince.
The effect of salmon plasma (SP) from Chinook salmon on proteolytic inhibition was investigated. SP was found to inhibit both cysteine and serine proteases as well as protease extracted from Pacific whiting muscle. SP was found to contain a 55kDa cysteine protease inhibitor through SDS-PAGE inhibitor staining. Freeze dried salmon plasma (FSP) and salmon plasma concentrated by ultrafiltration (CSP) were tested for their ability to inhibit autolysis in Pacific whiting surimi and salmon mince at concentrations of 0.25%, 0.5%, 1%, and 2%. Pacific whiting surimi autolysis was inhibited by an average of 89% regardless of concentration while inhibition of salmon mince autolysis increased with concentration (p<0.05). CSP performed slightly better than FSP at inhibiting salmon mince autolysis (p<0.05). Serine protease inhibition decreased when SP heated above 40°C but was stable across a broad NaCl and pH range. Cysteine protease inhibitors exhibited good temperature, NaCl, and pH stability.


Tailoring the ultrafast and nonlinear photonics of MXenes through elemental replacement.
Due to the outstanding electronic properties, unique chemical surface termination units and rich elemental compositions, MXenes have become promising candidates for the development of new generation optoelectronic devices. However, there is still a gap between advanced photonics applications and fundamental understanding of ultrafast carrier photo-physics dynamics and a nonlinear optical response in layered MXenes. Here, we present insight into the excited state relaxation processes and nonlinear optical response of few-layer Ti3CN and Ti3C2 nanosheets (NSs) via transient absorption spectroscopy and Z-scan measurements. Owing to similar structural compositions, the transient absorption and nonlinear absorption characteristics behave totally opposite. In addition, photo-induced bandgap renormalization and Pauli blocking phenomena exist in Ti3C2 and Ti3CN NSs, respectively. The element replacement may be a new strategy for tunable carrier kinetics and nonlinear optical response of MXenes. These research studies may provide insight into ultrafast carrier photo-physics dynamics as well as promote MXene-based advanced photonics and their applications in optoelectronic devices.


Emergency Department Query for Patient-Centered Approaches to Sexual Orientation and Gender Identity : The EQUALITY Study.
The Institute of Medicine and The Joint Commission recommend routine documentation of patients' sexual orientation in health care settings. Currently, very few health care systems collect these data since patient preferences and health care professionals' support regarding collection of data about patient sexual orientation are unknown. To identify the optimal patient-centered approach to collect sexual orientation data in the emergency department (ED) in the Emergency Department Query for Patient-Centered Approaches to Sexual Orientation and Gender Identity study. An exploratory, sequential, mixed-methods design was used first to evaluate qualitative interviews conducted in the Baltimore, Maryland, and Washington, DC, areas. Fifty-three patients and 26 health care professionals participated in the qualitative interviews. Interviews were followed by a national online survey, in which 1516 (potential) patients (244 lesbian, 289 gay, 179 bisexual, and 804 straight) and 429 ED health care professionals (209 physicians and 220 nurses) participated. Survey participants were recruited using random digit dialing and address-based sampling techniques. Qualitative interviews were used to obtain the perspectives of patients and health care professionals on sexual orientation data collection, and a quantitative survey was used to gauge patients' and health care professionals' willingness to provide or obtain sexual orientation information. Mean (SD) age of patient and clinician participants was 49 (16.4) and 51 (9.4) years, respectively. Qualitative interviews suggested that patients were less likely to refuse to provide sexual orientation than providers expected. Nationally, 154 patients (10.3%) reported that they would refuse to provide sexual orientation; however, 333 (77.8%) of all clinicians thought patients would refuse to provide sexual orientation. After adjustment for demographic characteristics, only bisexual patients had increased odds of refusing to provide sexual orientation compared with heterosexual patients (odds ratio, 2.40; 95% CI, 1.26-4.56). Patients and health care professionals have discordant views on routine collection of data on sexual orientation. A minority of patients would refuse to provide sexual orientation. Implementation of a standardized, patient-centered approach for routine collection of sexual orientation data is required on a national scale to help to identify and address health disparities among lesbian, gay, and bisexual populations.


Programs and procedures for a large x-ray safety program.
As radiation safety surveys of x-ray equipment are the responsibility of many health physicists, actual procedures vary among facilities. In an effort to share survey methodology of a large research institution, aspects of the x-ray safety program at Los Alamos National Laboratory are described. A definition of an intentional x-ray generating device is proposed in light of the absence or inadequacy of such a definition in applicable federal law and national consensus guidance documents. Regulations and procedures are discussed, and a sample database used in managing the Los Alamos program is presented.


Concerted regulation of mitochondrial and nuclear non-coding RNAs by a dual-targeted RNase Z.
The molecular roles of the dually targeted ElaC domain protein 2 (ELAC2) during nuclear and mitochondrial RNA processing in vivo have not been distinguished. We generated conditional knockout mice of ELAC2 to identify that it is essential for life and its activity is non-redundant. Heart and skeletal muscle-specific loss of ELAC2 causes dilated cardiomyopathy and premature death at 4 weeks. Transcriptome-wide analyses of total RNAs, small RNAs, mitochondrial RNAs, and miRNAs identified the molecular targets of ELAC2 in vivo We show that ELAC2 is required for processing of tRNAs and for the balanced maintenance of C/D box snoRNAs, miRNAs, and a new class of tRNA fragments. We identify that correct biogenesis of regulatory non-coding RNAs is essential for both cytoplasmic and mitochondrial protein synthesis and the assembly of mitochondrial ribosomes and cytoplasmic polysomes. We show that nuclear tRNA processing is required for the balanced production of snoRNAs and miRNAs for gene expression and that 3' tRNA processing is an essential step in the production of all mature mitochondrial RNAs and the majority of nuclear tRNAs.


Vascular alterations in apolipoprotein A-I amyloidosis (Leu75Pro). A case-control study.
Among hereditary amyloidoses, apolipoprotein A-I (APO A-I) amyloidosis (Leu75Pro) is a rare, autosomal dominant condition in which renal, hepatic, and testicular involvement has been demonstrated. To investigate vascular structural as well as functional alterations. In 131 carriers of the amyloidogenic Leu75Pro APO A-I mutation (mean age 52 + 16 years, 56 women) and in 131 subjects matched for age, sex, body mass index and clinic blood pressure (BP), arterial stiffness (pulse wave velocity, PWV) and carotid intima-media thickness (IMT) were measured. By definition no differences for age, sex, body mass index, and BP were observed. Meanmax IMT (Mmax-IMT) in the common (CC), bifurcation (BIF) and internal (ICA) carotid artery were comparable in the two groups. After adjustment for high-density lipoprotein cholesterol and renal function differences between the two groups, a lower meanmax-IMT was observed in APO A-I Leu75Pro mutation carriers than in controls (CC Mmax-IMT 0.87 ± 0.21 versus 0.93 ± 0.2 mm, p = 0.07; BIF Mmax-IMT 1.19 ± 0.48 versus 1.36 ± 0.46 mm, p = 0.025; ICA Mmax-IMT 0.9 ± 0.37 versus 1.02 ± 0.35 mm, p = 0.028). On the other hand, aortic stiffness was significantly greater in patients with APO A-I amyloidosis than controls (PWV 11.5 ± 2.9 and 10.7 ± 2.3 m/s, p < 0.05), even after adjusting for confounders. In carriers of the amyloidogenic Leu75Pro APO A-I mutation, a significant increase in arterial stiffness is observed; on the contrary, carotid artery IMT is comparable to that of control subjects. These results may add significant information to the clinical features of this rare genetic disorder.


Ketamine: A Practical Review for the Consultation-Liaison Psychiatrist.
Ketamine is a noncompetitive N-methyl-D-aspartate-receptor antagonist often used for sedation and management of acute agitation in general hospital settings. Many hospitals now include ketamine as part of their standard agitation protocol, and consultation-liaison psychiatrists frequently find themselves treating patients who have received ketamine, despite lack of clear recommendations for management. Conduct a nonsystematic narrative review regarding the use of ketamine for agitation and continuous sedation, including benefits and adverse psychiatric effects. Compare ketamine to more traditional agents of agitation control. Provide consultation-liaison psychiatrists with a summary of available knowledge and recommendations for managing patients receiving ketamine. A literature review was performed using PubMed, querying published articles from inception to March 2023 for articles related to use of ketamine for agitation or continuous sedation and side effects including psychosis and catatonia. A total of 37 articles were included. Ketamine was found to have multiple benefits, including shorter time to adequate sedation for agitated patients when compared to haloperidol ± benzodiazepines and unique advantages for continuous sedation. However, ketamine carries significant medical risks including high rates of intubation. Ketamine appears to induce a syndrome that mimics schizophrenia in healthy controls, and such effects are more pronounced and longer-lasting in patients with schizophrenia. Evidence regarding rates of delirium with ketamine for continuous sedation is mixed and requires further investigation before the agent is widely adopted for this purpose. Finally, the diagnosis of "excited delirium syndrome" and use of ketamine to treat this controversial syndrome warrants critical evaluation. Ketamine carries many potential benefits and can be an appropriate medication for patients with profound undifferentiated agitation. However, intubation rates remain high, and ketamine may worsen underlying psychotic disorders. It is essential that consultation-liaison psychiatrists understand the advantages, disadvantages, biased administration, and areas of limited knowledge regarding ketamine.


Light-regulated overexpression of an Arabidopsis phytochrome A gene in rice alters plant architecture and increases grain yield.
The phytochromes are a family of red/far-red light absorbing photoreceptors that control plant developmental and metabolic processes in response to changes in the light environment. We report here the overexpression of Arabidopsis thaliana PHYTOCHROME A (PHYA) gene in a commercially important indica rice variety (Oryza sativa L. Pusa Basmati-1). The expression of the transgene was driven by the light-regulated and tissue-specific rice rbcS promoter. Several independent homozygous sixth generation (T(5)) transgenic lines were characterized and shown to accumulate relatively high levels of PHYA protein in the light. Under both far-red and red light, PHYA-overexpressing lines showed inhibition of the coleoptile extension in comparison to non-transgenic seedlings. Furthermore, compared with non-transgenic rice plants, mature transgenic plants showed significant reduction in plant height, internode length and internode diameter (including differences in cell size and number), and produced an increased number of panicles per plant. Under greenhouse conditions, rice grain yield was 6-21% higher in three PHYA-overexpressing lines than in non-transgenic plants. These results demonstrate the potential of manipulating light signal-transduction pathways to minimize the problems of lodging in basmati/aromatic rice and to enhance grain productivity.


Evaluation of intranasal midazolam in children undergoing esophagogastroduodenoscopy.
Intravenous midazolam and opioids are used to produce conscious sedation in children undergoing esophagogastroduodenoscopy (EGD). However, children may experience significant fear and anxiety before receiving these medications, especially during separation from parents and during venipuncture. Intranasal administration of midazolam represents a noninvasive method of sedating children before anxiety-producing events. The objective of this study was to determine whether premedication with intranasal midazolam reduces stress and anxiety of separation from parents and of undergoing venipuncture, while maintaining adequate sedation during EGD. This was a prospective, randomized, double-blind study in 40 children, aged 2 to 12 years, who were undergoing EGD. Patients in group I were premedicated with intranasal placebo (0.9% NaCl) followed 10 minutes later by intravenous midazolam (0.05 mg/kg) and intravenous meperidine (1 mg/ kg). Patients in group II were premedicated with intranasal midazolam (0.2 mg/kg) followed by intravenous placebo (0.9% NaCl) and intravenous meperidine (1 mg/kg). Anxiolysis and sedation were scored by a blinded observer, who identified minor and major negative behaviors during four observation periods: intranasal drug administration, separation from parents, venipuncture, and EGD. Premedication with intranasal midazolam significantly reduced negative behaviors during separation from parents (p < 0.05); however, no difference between regimens was noted during venipuncture or EGD. Negative behaviors appeared to increase during administration of intranasal midazolam or placebo. Premedication with intranasal midazolam is effective in reducing negative behaviors during separation from parents, while it maintains sedation during the endoscopic procedure. The benefits of intranasal administration may be negated, however, by irritation, and discomfort caused by intranasal drug delivery.


Design considerations for massively parallel sequencing studies of complex human disease.
Massively Parallel Sequencing (MPS) allows sequencing of entire exomes and genomes to now be done at reasonable cost, and its utility for identifying genes responsible for rare Mendelian disorders has been demonstrated. However, for a complex disease, study designs need to accommodate substantial degrees of locus, allelic, and phenotypic heterogeneity, as well as complex relationships between genotype and phenotype. Such considerations include careful selection of samples for sequencing and a well-developed strategy for identifying the few "true" disease susceptibility genes from among the many irrelevant genes that will be found to harbor rare variants. To examine these issues we have performed simulation-based analyses in order to compare several strategies for MPS sequencing in complex disease. Factors examined include genetic architecture, sample size, number and relationship of individuals selected for sequencing, and a variety of filters based on variant type, multiple observations of genes and concordance of genetic variants within pedigrees. A two-stage design was assumed where genes from the MPS analysis of high-risk families are evaluated in a secondary screening phase of a larger set of probands with more modest family histories. Designs were evaluated using a cost function that assumes the cost of sequencing the whole exome is 400 times that of sequencing a single candidate gene. Results indicate that while requiring variants to be identified in multiple pedigrees and/or in multiple individuals in the same pedigree are effective strategies for reducing false positives, there is a danger of over-filtering so that most true susceptibility genes are missed. In most cases, sequencing more than two individuals per pedigree results in reduced power without any benefit in terms of reduced overall cost. Further, our results suggest that although no single strategy is optimal, simulations can provide important guidelines for study design.


Pharmacobiological management of hemostasis within clinical backgrounds via Ankaferd hemostat (Ankaferd blood stopper).
Ankaferd hemostat (Ankaferd blood stopper [ABS], Istanbul, Turkey) is a hemostatic agent affecting red blood cell-fibrinogen interactions. ABS has been traditionally used in Anatolia as a hemostatic agent for centuries. ABS contains a standardized combination of the plants namely Glycyrrhiza glabra, Thymus vulgaris, Alpinia officinarum, Vitis vinifera, and Urtica dioica. The hemostatic effect of ABS depends upon the quick promotion of a protein network, particularly fibrinogen gamma, in relation to the erythrocyte aggregation. The aim of this review is to indicate pharmacobiological basis and clinical backgrounds of ABS. Current perspective for using ABS is to provide hemostasis and accelerating wound healing particularly in cases which are difficult to manage. Future controlled trials are needed to elucidate the actions of ABS with in hemostasis, antithrombotic, anti-inflammatory, anti-infective, antifungal, and anti-oxidative effects.


Signaling between somatic follicle cells and the germline patterns the egg and embryo of Drosophila.
In Drosophila, specification of the embryonic body axes requires signaling between the germline and the somatic follicle cells. These signaling events are necessary to properly localize embryonic patterning determinants in the egg or eggshell during oogenesis. There are three maternal patterning systems that specify the anterior-posterior axis, and one that establishes the dorsal-ventral axis. We will first review oogenesis, focusing on the establishment of the oocyte and nurse cells and patterning of the follicle cells into different subpopulations. We then describe how two coordinated signaling events between the oocyte and follicle cells establish polarity of the oocyte and localize the anterior determinant bicoid, the posterior determinant oskar, and Gurken/epidermal growth factor (EGF), which breaks symmetry to initiate dorsal-ventral axis establishment. Next, we review how dorsal-ventral asymmetry of the follicle cells is transmitted to the embryo. This process also involves Gurken-EGF receptor (EGFR) signaling between the oocyte and follicle cells, leading to ventrally-restricted expression of the sulfotransferase Pipe. These events promote the ventral processing of Spaetzle, a ligand for Toll, which ultimately sets up the embryonic dorsal-ventral axis. We then describe the activation of the terminal patterning system by specialized polar follicle cells. Finally, we present open questions regarding soma-germline signaling during Drosophila oogenesis required for cell identity and embryonic axis formation.


Prediction of lung function using handgrip strength in healthy young adults.
Positive association between physical activity and spirometry has been reported to be possibly attributed to handgrip strength (HGS), particularly in the elderly. However, the nature of the association between HGS and lung function in young adults is still unclear. This study investigated the prediction of lung function using HGS in young adults. A cross-sectional analytical study was carried out on four hundred (400) apparently healthy medical students who are aged 16-30 years. Handgrip strength (dominant and nondominant) and lung function (FEV1 , FVC and PEFR) of these students were assessed using Jamar dynamometer and a portable spirometer, respectively. Data were analyzed using inferential statistics. Independent t-test showed that the mean values of HGS and lung function of the males were significantly higher than the females (P < 0.0005). The relationship between HGS and lung function indices was significant (P < 0.0005) in all the participants but strongest for FEV1 (r = 0.64). The regression analysis showed that in addition to gender and height, HGS was a significant (P < 0.0005) predictor of lung function. Regression equations were also proposed for the prediction of these lung function indices using HGS, gender and height. This study is the first to report HGS as a significant predictor of pulmonary function in healthy young adults living in a low-resource country. Hence, its use could enhance medical practice in being an indicator of lung function status in healthy young adults.