Sodium-dependent nucleoside transport in choroid plexus from rabbit. Evidence for a single transporter for purine and pyrimidine nucleosides.
The overall goal of this study was to determine the mechanisms by which nucleosides are transported in choroid plexus. Choroid plexus tissue slices obtained from rabbit brain were depleted of ATP with 2,4-dinitrophenol. Uridine and thymidine accumulated in the slices against a concentration gradient in the presence of an inwardly directed Na+ gradient. The Na(+)-driven uptake of uridine and thymidine was saturable with Km values of 18.1 +/- 2.0 and 13.0 +/- 2.3 microM and Vmax values of 5.5 +/- 0.3 and 1.0 +/- 0.2 nmol/g/s, respectively. Na(+)-driven uridine uptake was inhibited by naturally occurring ribo- and deoxyribonucleosides (adenosine, cytidine, and thymidine) but not by synthetic nucleoside analogs (dideoxyadenosine, dideoxycytidine, cytidine arabinoside, and 3'-azidothymidine). Both purine (guanosine, inosine, formycin B) and pyrimidine nucleosides (uridine and cytidine) were potent inhibitors of Na(+)-thymidine transport with IC50 values ranging between 5 and 23 microM. Formycin B competitively inhibited Na(+)-thymidine uptake and thymidine trans-stimulated formycin B uptake. These data suggest that both purine and pyrimidine nucleosides are substrates of the same system. The stoichiometric coupling ratios between Na+ and the nucleosides, guanosine, uridine, and thymidine, were 1.87 +/- 0.10, 1.99 +/- 0.35, and 2.07 +/- 0.09, respectively. The system differs from Na(+)-nucleoside co-transport systems in other tissues which are generally selective for either purine or pyrimidine nucleosides and which have stoichiometric ratios of 1. This study represents the first direct demonstration of a unique Na(+)-nucleoside co-transport system in choroid plexus.


Medically ready for discharge: A multisite "point-in-time" assessment of hospitalized patients.
Time spent awaiting discharge after the acute need for hospitalization has resolved is an important potential contributor to hospital length of stay (LOS). To measure the prevalence, impact, and context of patients who remain hospitalized for prolonged periods after completion of acute care needs. We conducted a cross-sectional "point-in-time" survey at each of 15 academic US hospitals using a structured data collection tool with on-service acute care medicine attending physicians in fall 2022. Primary outcomes were number and percentage of patients considered "medically ready for discharge" with emphasis on those who had experienced a "major barrier to discharge" (medically ready for discharge for ≥1 week). Estimated LOS attributable to major discharge barriers, contributory discharge needs, and associated hospital characteristics were measured. Of 1928 patients sampled, 35.0% (n = 674) were medically ready for discharge including 9.8% (n = 189) with major discharge barriers. Many patients with major discharge barriers (44.4%; 84/189) had spent a month or longer medically ready for discharge and commonly (84.1%; 159/189) required some form of skilled therapy or daily living support services for discharge. Higher proportions of patients experiencing major discharge barriers were found in public versus private, nonprofit hospitals (12.0% vs. 7.2%; p = .001) and county versus noncounty hospitals (14.5% vs. 8.8%; p = .002). Patients experience major discharge barriers in many US hospitals and spend prolonged time awaiting discharge, often for support needs that may be outside of clinician control.


Assessment of trace metal contamination in the sea cucumber (Holothuria tubulosa) and sediments from the Dardanelles Strait (Turkey).
This study was performed to determine the concentrations of some trace metals (Cd, Cu, Pb, Ni, Zn, and Fe) in Holothuria tubuosa (Gmelin, 1788) belonging to Echinoderm species and in sediments that they live at three different stations (Gelibolu, Umur Bey/Lapseki, and Dardanos) on Dardanelles Strait between April 2013 and March 2014. The mean trace metal concentrations determined in H. tubulosa and sediment were as follows: Cd 0.18 mg/kg, Cu 2.43 mg/kg, Pb 2.09 mg/kg, Ni 14.58 mg/kg, Zn 16.86 mg/kg, and Fe 73.46 mg/kg and Cd 0.70 mg/kg, Cu 5.03 mg/kg, Pb 14.57 mg/kg, Ni 27.15 mg/kg, Zn 54.52 mg/kg, and Fe 3779.9 mg/kg, respectively. It was detected that the statistical difference between trace metals determined seasonally in muscle tissue of H. tubulosa was significant (p > 0.05). As a result of the study, it was detected that H. tubulosa is a bioindicator species in determining Ni trace metal in sediment. The results were compared to the limit values of National and International Food Safety, and it was detected that Cd and Ni concentrations measured in sediment were above LEL of Ni and Cd concentrations according to Sediment Quality Guidelines.


Coordinator teaches staff to operate equipment, handles malfunctioning equipment.
Based on the data collection, a recommendation was made to the OR committee to develop a new full-time position of equipment coordinator. Committee members supported the recommendation, and further recommended that a backup person be trained as an assistant equipment coordinator to cover when the equipment coordinator was not available.


Translationally Controlled Tumor Protein-Mediated Stabilization of Host Antiapoptotic Protein MCL-1 Is Critical for Establishment of Infection by Intramacrophage Parasite Leishmania donovani.
In the early phase of infection, the intramacrophage pathogen Leishmania donovani protects its niche with the help of the antiapoptotic protein myeloid cell leukemia-1 (MCL-1). Whether Leishmania could exploit MCL-1, an extremely labile protein, at the late phase is still unclear. A steady translational level of MCL-1 observed up to 48 h postinfection and increased caspase-3 activity in MCL-1-silenced infected macrophages documented its importance in the late hours of infection. The transcript level of MCL-1 showed a sharp decline at 6 h postinfection, and persistent MCL-1 expression in cyclohexamide-treated cells negates the possibility of de novo protein synthesis, thereby suggesting infection-induced stability. Increased ubiquitination, a prerequisite for proteasomal degradation of MCL-1, was also found to be absent in the late hours of infection. Lack of interaction with its specific E3 ubiquitin ligase MULE (MCL-1 ubiquitin ligase E3) and specific deubiquitinase USP9X prompted us to search for blockade of the ubiquitin-binding site in MCL-1. To this end, TCTP (translationally controlled tumor protein), a well-known binding partner of MCL-1 and antiapoptotic regulator, was found to be strongly associated with MCL-1 during infection. Phosphorylation of TCTP, a requirement for MCL-1 binding, was also increased in infected macrophages. Knockdown of TCTP decreased MCL-1 expression and short hairpin RNA-mediated silencing of TCTP in an infected mouse model of visceral leishmaniasis showed decreased parasite burden and induction of liver cell apoptosis. Collectively, our investigation revealed a key mechanism of how L. donovani exploits TCTP to establish infection within the host.


Unpacking the Internalized Homonegativity-Health Relationship: How the Measurement of Internalized Homonegativity and Health Matter and the Contribution of Religiousness.
Internalized homonegativity (IH) is widely recognized to negatively influence the health of lesbian, gay, bisexual, and queer/questioning (LGBQ +) individuals. It is not clear, however, the role that religiousness may play in the relationship between IH and health or how differing conceptualizations of IH or health may influence this relationship. We conducted a multi-level meta-analysis of 151 effect sizes from 68 studies to examine the relationship between IH and health as well as what may moderate this relationship. Results suggested that IH was consistently and negatively related to health (r = - .28). Analyses suggest that IH was most strongly related to mental health and well-being, evidencing a relatively weaker (though still negative) relationship with physical or sexual health. Analyses of different ways to measure IH suggest that scales that conceptualize IH to include distress (e.g., the Internalized Homonegativity Inventory) report stronger relationships with health than scales that conceptualize IH orthogonally from distress (e.g., the Internalized Homonegativity scale of the Lesbian, Gay, Bisexual Identity Salience Scale [LGBIS]). We failed to find significant moderation effects for religiousness, though it was hard to evaluate this relationship due to the poor quality of most measures of religiousness. We encourage researchers to use measures of IH that conceptualize IH orthogonally from distress (e.g., the LGBIS) and to use more nuanced measures of religiousness (e.g., religious belief, religious activity). We also encourage researchers to focus on how IH impacts less-often studied measures of health (e.g., physical health, suicidality).


Nuclear Overhauser effects in linear peptides. A low-temperature 500 MHz study of Met-enkephalin.
Met5-enkephalin was studied in 1 mM solutions in 2H2O at room temperature and in a cryoprotective mixture (DMSOd6/2H2O, mole fraction of DMSO 0.49) in the temperature range 265-298 K. Small positive effects were observed between the ortho and meta protons of Tyr in aqueous solution at room temperature. Intraresidue effects can be made strong and negative by increasing the viscosity of the medium with a combination of cryoprotective mixtures and low temperatures. The use of mixtures with properties very close to water is very promising for conformational studies of enkephalins and of other small linear peptides.


Clinical attachment level gain of lasers in scaling and root planing of chronic periodontitis: a network meta-analysis of randomized controlled clinical trials.
The objective of this study was to evaluate the clinical attachment level (CAL) gain of Er:YAG, Er,Cr; YSGG, Nd:YAG; and diode laser (DL) as monotherapy or adjunctive to scaling and root planing (SRP) of chronic periodontitis by network meta-analysis (NMA). Randomized controlled clinical trials (RCTs) about lasers applied in SRP of chronic periodontitis were searched from PubMed, Cochrane Library, Web of Science, Ovid, Science Direct, Wan Fang, and China National Knowledge Infrastructure (CNKI) databases up to September 2018 and from references of selected full-texts and related reviews. Standard mean differences and 95% confidence intervals were counted for CAL gain. The random effects NMA were performed in STATA software. There were 25 RCTs about CAL gain at 3 and/or 6 months after lasers were applied in SRP. No inconsistency was detected. Er:YAG as monotherapy gained significantly more CAL at 3 months than did SRP; no significant differences were detected among other comparisons. In terms of CAL gain at 3 months, the ranking result from best to worst was as follows: Er:YAG as monotherapy, DL adjunctive to SRP, Er:YAG adjunctive to SRP, Er,Cr;YSGG as monotherapy, Nd:YAG adjunctive to SRP, and SRP. In terms of CAL gain at 6 months, the ranking result from best to worst was as follows: DL adjunctive to SRP, Nd:YAG adjunctive to SRP, SRP, Er:YAG adjunctive to SRP, and Er:YAG as monotherapy. Laser-assisted periodontal treatment could be superior to SRP alone and could serve as a good adjunctive treatment tool.


Boston brace correction in idiopathic scoliosis: a biomechanical study.
To analyze Boston brace biomechanics, pressure measurements and finite element simulations were done on 12 adolescent idiopathic scoliosis patients. The aim was to analyze the Boston brace effectiveness using a finite element model and experimental measurements. There are not very many biomechanical studies of Boston brace effectiveness, and its biomechanical action is not completely understood. This study was performed on 12 girls with scoliosis treated with the Boston brace system. The experimental protocol was composed of the acquisition of two sets of multiplanar radiographs with and without brace followed by the pressure acquisition at the brace-torso interface. A personalized finite element modeling of the trunk was generated from the 3D reconstruction of the patient's geometry. The brace treatment was simulated by the application of equivalent forces calculated from the pressure measurements. Two Boston brace force patterns were defined from the pressure measurements. The first one consisted of high right thoracic forces of 31-113 N, lumbar forces less than 47 N, and included a left thoracic extension working as a counter pad. The second one consisted of low thoracic forces less than 20 N, lumbar forces up to 70 N, without left thoracic extension. The simulations showed that the passive forces only produced a coronal Cobb angle correction up to 9 degrees, whereas real correction was up to 16 degrees. High thoracic pads reduced more effectively both thoracic and lumbar scoliotic curves than lumbar pads only. The study suggests that mechanisms other than brace pads produce correction and contribute to the force equilibrium within the brace.


Introducing Edna: A trainee chatbot designed to support communication about additional (secondary) genomic findings.
To support informed decision-making about reanalysis of clinical genomic data for risk of preventable conditions ('additional findings') by developing a chatbot (electronic genetic resource, 'eDNA'). Interactions in pre-test genetic counseling sessions (13.5 h) about additional findings were characterized using proponent, thematic and semantic analyses of transcripts. We then wrote interfaces to draw supplementary data from external genetics applications. To create Edna, this content was programmed using a chatbot framework which interacts with patients via speech-to-text. Conditions, terms, explanations of concepts, and key factors to consider in decision making were all encoded into chatbot conversations emulating counseling session flows. Patient agency can be enhanced by prompted consideration of the personal and familial implications of testing. Similarly, health literacy can be broadened through explanation of genetic conditions and terminology. Novel aspects include sentiment analysis and collection of family history. Medical advice and the impact of existing genetic conditions were deemed inappropriate for inclusion. Edna's successful development represents a movement towards accessible, acceptable and well-supported digital health processes for patients to make informed decisions for additional findings. Edna complements genetic counseling by collecting and providing genomic information before or after pre-test consultations.


The synthetic gestagen levonorgestrel directly affects gene expression in thyroid and pituitary glands of Xenopus laevis tadpoles.
The synthetic gestagen levonorgestrel (LNG) was previously shown to perturb thyroid hormone-dependent metamorphosis in Xenopus laevis. However, so far the mechanisms underlying the anti-metamorphic effects of LNG remained unknown. Therefore, a series of in vivo and ex vivo experiments was performed to identify potential target sites of LNG action along the pituitary-thyroid axis of X. laevis tadpoles. Prometamorphic tadpoles were treated in vivo with LNG (0.01-10nM) for 72h and brain-pituitary and thyroid tissue was analyzed for marker gene expression. While no treatment-related changes were observed in brain-pituitary tissue, LNG treatment readily affected thyroidal gene expression in tadpoles including decreased slc5a5 and iyd mRNA expression and a strong induction of dio2 and dio3 expression. When using an ex vivo organ explant culture approach, direct effects of LNG on both pituitary and thyroid gland gene expression were detecTable Specifically, treatment of pituitary explants with 10nM LNG strongly stimulated dio2 expression and concurrently suppressed tshb expression. In thyroid glands, ex vivo LNG treatment induced dio2 and dio3 mRNA expression in a thyrotropin-independent manner. When thyroid explants were cultured in thyrotropin-containing media, LNG caused similar gene expression changes as seen after 72h in vivo treatment including a very strong repression of thyrotropin-induced slc5a5 expression. Concerning the anti-thyroidal activity of LNG as seen under in vivo conditions, our ex vivo data provide clear evidence that LNG directly affects expression of genes important for thyroidal iodide handling as well as genes involved in negative feedback regulation of pituitary tshb expression.


Cell-mediated immunity to respiratory virus infections.
The mucosal surfaces of the lungs pose tremendous problems for an immune system charged with maintaining a sterile pulmonary environment. Despite these problems, the immune system is effective at controlling most pulmonary infections. Over the past few years significant progress has been made in our understanding of how adaptive (humoral and cellular) immunity is able to control infections in the respiratory tract. Recent advances include the identification of effector memory T-cell populations in the lungs and an appreciation for the role of cytokines in regulating memory T-cell pools.


Flow karyotype analysis and fluorescence-activated sorting of Burkitt-lymphoma-associated translocation chromosomes.
Flow karyotype analysis was performed to assess the feasibility of fluorescence-activated sorting of Burkitt lymphoma (BL)-associated translocation chromosomes. The typical 14q+ chromosome in the t(8;14) and the two "variant translocations", the 2q+ in the t(2;8) and the small 22q- in the t(8;22), could be identified as single peaks within the flow karyotypes of metaphase chromosomes isolated from several different BL-lines for each translocation. The translocation chromosomes could be separated with a high degree of purity and in quantities suitable for biochemical analysis. The same analytical and preparative technique was also successfully applied to the identification and sorting of the Philadelphia (Ph1) chromosome in a 9;22 translocation-carrying CML-derived line and a familial (11;22) translocation.


Membrane transducing activity of recombinant Hoxc8 protein and its possible application as a gene delivery vector.
In order to examine whether the Hoxc8 protein can deliver nucleic acid into mammalian cells, we designed several Hoxc8-derived recombinant proteins to be synthesized as glutathione S-transferase (GST) fused forms in E. coli (GST-Hoxc8(1-242), containing a full length of Hoxc8; GST-Hoxc8(152-242), possessing a deletion of the acidic N-terminus of Hoxc8; GST-Hoxc8(149-208), which contained the homeodomain only). After labeling these proteins with Oregon 488, we examined their membrane transduction ability under the fluorescence microscope and verified that all three proteins showed similar transduction efficiency. The ability of the proteins to form in vitro protein-DNA complexes was analyzed on agarose gel; both GST-Hoxc8(1-242) and GST-Hoxc8(149-208) formed complexes. In contrast, the GST-Hoxc8(152-242) protein did not form a complex. The GST-Hoxc8(149-208) protein formed a complex with DNA at a mass ratio of 1ú1 (DNAúprotein), and GST-Hoxc8(1-242) formed a complex at a mass ratio of 1ú5. When the DNA (pDsRed1-C1) and protein complexes were added to culture media containing mammalian cells, the cells uptook the complexes, which was indicated by red fluorescence expression under the fluorescent microscope. These results indicate that recombinant Hoxc8 derivatives that harbor a homeodomain are able to traverse the mammalian cellular membrane. DNA that is bound to the recombinant derivatives can be carried across the membrane as well. This process could be applied in the development of a useful delivery vector for gene therapy in the future.


The Association Between Essential Metal Element Mixture and Sleep Quality in Chinese Community-Dwelling Older Adults.
Previous studies have related single essential metal elements (EMEs) to sleep quality among older adults, however, the association of the EME mixture with sleep quality remained poorly understood. This study aimed to investigate the relationships between single EMEs and the EME mixture and sleep quality in older adults living in Chinese communities. This study consisted of 3957 older adults aged 60 years or over. Urinary concentrations of cobalt (Co), vanadium (V), selenium (Se), molybdenum (Mo), strontium (Sr), calcium (Ca), and magnesium (Mg) were detected using inductively coupled plasma mass spectrometry. Sleep quality was evaluated using Pittsburgh Sleep Quality Index (PSQI). The associations of single EMEs and EME mixture with sleep quality were assessed using logistic regression and Bayesian kernel machine regression (BKMR) models, respectively. Adjusted single-element logistic regression models showed that Mo (OR = 0.927, 95%CI:0.867-0.990), Sr (OR = 0.927, 95%CI:0.864-0.994), and Mg (OR = 0.934, 95%CI:0.873-0.997) were negatively related to poor sleep quality. BKMR models exhibited similar results. Also, higher levels of the EME mixture in urine were inversely related to the odds of poor sleep quality after adjustment for covariates, and Mo had the largest conditional posterior inclusion probability (condPIP) value in the mixture. Mo, Sr, and Mg were negatively related to poor sleep quality, separately and as the mixture. The EME mixture in urine was associated with decreased odds of poor sleep quality in older adults, and Mo was the greatest contributor within the mixture. Additional cohort research is warranted to clarify the relationship of multiple EMEs with sleep quality.


Sb(iii)/Gum Arabic composite as a new natural-based environmentally green catalyst for the one-pot pseudo-four-component synthesis of 2H-indazolo[2,1-b] phthalazinetriones.
Sb(iii)-Gum Arabic composite as a unique natural-based and nontoxic catalyst was synthesized and characterized by FT-IR, XRD, TGA, ICP, BET, EDX and mapping. A four-component reaction of phthalic anhydride, hydrazinium hydroxide, aldehyde, and dimedone has been carried out in the presence of Sb(iii)/Gum Arabic composite to synthesise 2H-indazolo[2,1-b] phthalazine triones. The advantages of the present protocol are the appropriate reaction times, eco-friendly nature and high yields.


[KTP laser coagulation for xanthelasma palpebrarum].
Xanthelasma palpebrarum is a benign skin disorder of unknown etiology appearing as one or multiple yellowish plaques around the eyelids. Hyperlipidemia, thyroid dysfunction, and diabetes mellitus are possible pathogenetic triggers. Removal of xanthelasma is desired for aesthetic reasons. While there are many treatment options, search continue for newer methods with less scarring and post-inflammatory hyperpigmentation. We employed for the first time KTP laser (532 nm) for the treatment of 14 patients with xanthelasma palpebrarum on 33 eyelids. More than 70% of the patients tolerated KTP laser irradiation without any analgesia. 85.7% showed respectable reduction of xanthelasma palpebrarum after one to three treatment sessions without side effects. KTP laser irradiation represents a minimally invasive, safe and effective treatment option for the reduction of xanthelasma palpebrarum without undesired side effects.


NMDA receptor diversity in the cerebellum: identification of subunits contributing to functional receptors.
Recent studies of N-methyl-D-aspartate (NMDA) receptors have led to the suggestion that there are two distinct classes of native NMDA receptors, identifiable from their single-channel conductance properties. 'High-conductance' openings arise from NR2A- or NR2B-containing receptors, and 'low-conductance' openings arise from NR2C- or NR2D-containing receptors. In addition, the low-conductance channels show reduced sensitivity to block by Mg2+. The readily identified cell types and simple architecture of the cerebellum make it an ideal model system in which to determine the contribution of specific subunits to functional NMDA receptors. Furthermore, mRNA for all of these four NR2 subunits are represented in this brain region. We have examined NMDA channels in Purkinje cells, deep cerebellar nuclei (DCN) neurons and Golgi cells. First we find that NR2D-containing NMDA receptors give rise to low-conductance openings in cell-attached recordings from Purkinje cells. The characteristic conductance of these events cannot, therefore, be ascribed to patch excision. Second, patches from some DCN neurons exhibit mixed populations of high- and low-conductance openings. Third, Golgi cells also exhibit a mixed population of high- and low-conductance NMDA receptor openings. The features of these low-conductance openings are consistent with the presence of NR2D-containing NMDA receptors, as suggested by in situ hybridization data. On the other hand the existence of high-conductance channels, with properties typical of NR2B-containing receptors, was not expected. Our results provide new evidence about the subunit composition of NMDA receptors in identified cerebellar cells, and suggest that examination of single-channel properties is a potentially powerful approach for determining the possible subunit composition of native NMDA receptors.


Role of activity-dependent BDNF expression in hippocampal-prefrontal cortical regulation of behavioral perseverance.
Activity-dependent gene transcription, including that of the brain-derived neurotrophic factor (Bdnf) gene, has been implicated in various cognitive functions. We previously demonstrated that mutant mice with selective disruption of activity-dependent BDNF expression (BDNF-KIV mice) exhibit deficits in GABA-mediated inhibition in the prefrontal cortex (PFC). Here, we show that disruption of activity-dependent BDNF expression impairs BDNF-dependent late-phase long-term potentiation (L-LTP) in CA1, a site of hippocampal output to the PFC. Interestingly, early-phase LTP and conventional L-LTP induced by strong tetanic stimulation were completely normal in BDNF-KIV mice. In parallel, attenuation of activity-dependent BDNF expression significantly impairs spatial memory reversal and contextual memory extinction, two executive functions that require intact hippocampal-PFC circuitry. In contrast, spatial and contextual memory per se were not affected. Thus, activity-dependent BDNF expression in the hippocampus and PFC may contribute to cognitive and behavioral flexibility. These results suggest distinct roles for different forms of L-LTP and provide a link between activity-dependent BDNF expression and behavioral perseverance, a hallmark of several psychiatric disorders.


Phylogenetic distribution of aromatase and other androgen-converting enzymes in the central nervous system.
Metabolism of [3H]androstenedione was studied in brain tissue homogenates of opossum, bird, snake, sea turtle, urodele amphibian, teleost, shark, skate, hagfish, and lobster. Estrone, 17 beta-estradiol, or 17 alpha-estradiol was formed by central neural tissues of all species, with the exception of the opossum, hagfish, and lobster. Aromatase activity was concentrated in the forebrain, although some estrogen was synthesized by mid- or hindbrain homogenates of two lower vertebrates (teleost and skate) and the newly hatched chick. 5 alpha-Androstanedione (5 alpha-androstane-3,17-dione) or 5 alpha-dihydrotestosterone were products of metabolism in several nonmammalian vertebrates and in the invertebrate central nervous system also. 5 alpha-Reductase was found in all major brain divisions. These and previously reported comparative studies indicate that the ability to aromatize and otherwise transform androgen substrates is a primitive characteristic of the brain that has been widely conserved phylogenetically.


Chronic hypernatremia from a congenital defect in osmoregulation of thirst and vasopressin.
An infant with microcephaly and delayed development was found to have chronic asymptomatic hypernatremia. Computerized brain tomography disclosed dysplasia of the midline structures, septum pellucidum and corpus collosum. Evaluation revealed defective osmoregulation, hypothalamic hypothyroidism, and hypogonadotropinism. He showed no desire to drink at plasma osmolalities over 330 mOsm/kg. His plasma vasopressin levels (less than or equal to 1.4 pg/ml) were inappropriately low relative to his high levels of plasma osmolality (greater than or equal to 310 mOsm/kg), which might be accounted for by either deficient neurohypophyseal vasopressin stores or disturbance of the hypothalamic osmoreceptors governing vasopressin. The first possibility was ruled out by demonstrating normal vasopressin response (167 pg/ml) to nonosmotic (emetic) stimulation. Under baseline conditions, his urine was concentrated up to 747 mOsm/kg and urine volume was low. With water loading, maximal water diuresis developed (urine osmolality 68 mOsm/kg), but his plasma osmolality remained in the hyperosmolar range (312 mOsm/kg). Treatment with a vasopressin analogue, desamino-D-arginine vasopressin, and forced hydration restored plasma osmolality and plasma sodium to normal. These findings indicate a severe defect in the hypothalamic osmoreceptors controlling thirst and vasopressin secretion with normal vasopressin stores and preserved vasopressin responsiveness to nonosmotic stimuli. To our knowledge, this report provides the first documentation of selective osmoreceptor defect in conjunction with congenital dysplasia of midline brain structures.


Regional homogeneity of spontaneous brain activity in adult patients with obsessive-compulsive disorder before and after cognitive behavioural therapy.
Cognitive behavioural therapy (CBT) is an effective treatment for obsessive-compulsive disorder (OCD). Several neuroimaging studies have explored alterations of brain function in OCD patients as they performed tasks after CBT. However, the effects of CBT on the neural activityin OCD during rest remain unknown. Therefore, we investigated changes in regional homogeneity (ReHo) in OCD patients before and after CBT. Twenty-two OCD patients and 22 well-matched healthy controls participated in the resting-state functional magnetic resonance imaging scans. We compared differences in ReHo between the OCD and control groups before treatment and investigated the changes of ReHo in 17 OCD patients who responded to CBT. Compared to healthy controls, OCD patients exhibited higher ReHo in the right orbitofrontal cortex (OFC), bilateral middle frontal cortex, right precuneus, left cerebellum, and vermis, as well as lower ReHo in the bilateral caudate, right calcarine, right posterior cingulate cortex, and right middle temporal cortex. Along with the clinical improvement in OCD patients after CBT, we found decreased ReHo in the right OFC, bilateral middle frontal cortex, left cerebellum and vermis, and increased ReHo in the left caudate. Improvement of OCD symptoms was significantly correlated with the changed ReHo in the right OFC and left cerebellum. Although these findings are preliminary and need to be replicated in larger samples, they indicate the presence of abnormal spontaneous brain activity of the prefrontal-striatal-cerebellar circuit in OCD patients, and provide evidence that CBT can selectively modulate the spontaneous brain activity of this circuit in OCD patients.


Endogenous endophthalmitis caused by Streptococcus suis infection: a case report.
Streptococcus suis (S. suis) is an emerging zoonotic human pathogen, which commonly causes meningitis and sepsis. Ocular infections associated with S. suis infection are very rare. Herein, we reported a rare case of a man who developed endophthalmitis complicated by meningitis following S. suis infection. A 48-year-old male with a fever, loss of vision in the right eye, slight headache, and hearing loss in the right ear was admitted to our hospital. A comprehensive ophthalmological examination suggested endophthalmitis. The metagenomics next-generation sequencing (mNGS) results of pathogenic microorganisms from vitreous and cerebrospinal fluid samples revealed that the causative pathogen was S. suis, which was further confirmed by the bacterial culture of the vitreous sample. Subsequently, the patient received phacoemulsification and vitrectomy, combined with silicone oil tamponade, as well as local and systemic anti-infective therapy, after which his condition significantly improved. Despite the low incidence rate of endophthalmitis caused by S. suis, clinicians should be aware of relevant clinical manifestations, especially for patients with neurological symptoms and risk factors for S. suis infection. The next-generation sequencing is efficient for etiological diagnosis of pathogenic microorganisms.


Effect of a final alcohol rinse on sealer coverage of obturated root canals.
An endodontic sealer is required to provide an apical and coronal seal in root canals. No previously reported methods of sealer placement completely coated canal walls. The purpose of this study was to dehydrate canal walls with alcohol and determine if this permitted better sealer coverage. Forty single-rooted teeth were prepared and divided into 4 groups of 10 teeth according to method of sealer placement and final irrigant: (a) by lentulo spiral and (b) file NaOCl, (c) lentulo and alcohol, and (d) file and alcohol. After canal preparation, groups 1 and 2 were irrigated with 1 ml of NaOCl and dried with paper points. Groups 3 and 4 were irrigated with 1 ml of alcohol and dried. AH26 (0.04 ml) dyed with charcoal black was applied to the apical end of the file or lentulo and placed in the canal to coat the walls. Gutta-percha without additional sealer was laterally condensed. Teeth were cleared and evaluated for location and amount of sealer visible. Data were evaluated by Kruskal-Wallis and analysis of variance. The results showed (a) no significant differences among the four groups, (b) all areas had sealer present, (c) the coronal third had > 50% sealer coverage in nearly all teeth, and (d) the middle and apical thirds showed the most variability in sealer coverage.


Standard operating procedures approach for the implementation of the evidence-based dentistry concept in dental practice.
Evidence-based dentistry is a concept that when applied to clinical practice may improve the quality of dental treatment. However, dentists' reluctance to change their behavior may be a barrier to the implementation of the process. The main purpose of this study was to demonstrate that standard operating procedures (SOPs) may help dentists to apply scientific evidence to their dental practice. SOPs are written instructions on how to execute some specific tasks. A flowchart model demonstrated how an ordinary clinical procedure (composite restoration) can be performed using evidence-based information to support each executed step. Implementing the model into daily practice is straightforward, and the results are accessible to the whole dental team. In addition, the flowchart can be regularly updated with high-quality dental literature such as systematic reviews of randomized controlled trials and randomized controlled trials. This proposed model may help to bridge the gap between research and clinical dental practice by serving as a practical tool to improve the knowledge of dental practitioners and the quality of treatment.


Variable susceptibility to NK activity of cloned cell lines derived from a primary rat rhabdomyosarcoma: relationship to metastatic potential.
In vitro cloned lines derived from a primary nickel-induced rat rhabdomyosarcoma exhibited diverse levels of susceptibility to spontaneous NK activity. The presence of NK target structures was revealed by competition assays on all cloned cell lines, and the NK susceptibility of the tumour lines varied according to their osmotic fragility. Tumour cell lines derived from metastatic lung nodules presented similar NK susceptibilities to cells originating from the primary tumour. However, cloned cell lines differed in their capacity to form lung colonies after i.v. injection, and in their potential for invading lungs after s.c. primary tumour development. No correlation was found between lung colonization potential and NK resistance. Studies of the correlation between metastatic potential and NK sensitivity revealed that (1) all the NK resistant tumour cells were highly metastatic; (2) NK susceptible tumour cells could be either highly or weakly metastatic. Therefore, highly metastatic tumour cells could be either resistant or susceptible to NK lysis. We conclude that the property of resistance to NK contributes to a high metastatic potential. However, other properties could counterbalance and finally prevail over NK susceptibility thus enabling NK susceptible cell lines to be also highly metastatic.


Flutrer-like P waves in a case of atrioventricular reciprocating tachycardia.
Typical atrial flutter is characterized by its sawtooth flutter wave in leads II, III, aVF, and V1. Atrioventricular reciprocating tachycardia is characterized by its small retrograde P wave after completion of QRS complex, where sawtooth flutter-like P waves are rarely seen in the electrocardiogram during atrioventricular reciprocating tachycardia. We report on a 62-year-old patient who presented the characteristic sawtooth flutter-like P waves in the electrocardiogram during attack of supraventricular tachycardia. By electrophysiologic study, the mechanism of his supraventricular tachycardia was atrioventricular reciprocating tachycardia using the left posterior lateral concealed accessory pathway for retrograde conduction. The accessory pathway was successfully ablated by radiofrequency ablation therapy.


Diagnosing lysosomal storage disorders: Fabry disease.
Fabry disease (FD) is an X-linked lysosomal storage disorder due to deficiency of alpha galactosidase A (GLA). Progressive, intralysosomal accumulation of neutral glycosphingolipids in endothelial cells and podocytes leads to multi-organ involvement in affected males and to a lesser extent in affected females. Diagnosis of FD is based on GLA analysis in leukocytes or dried blood spots (DBS) in FD males while GLA activities may be within the normal range in FD females. The advent of fluorometric and mass spectrometry methods for enzyme analysis in DBS has simplified the diagnostic approach for FD males, facilitating high-throughput screening of at risk populations and newborn infants. However, the diagnostic mainstay for FD females remains molecular analysis of the GLA gene. The following unit will provide the detailed analytical protocol for measurement of GLA activity in DBS using tandem mass spectrometry.


Abiotic factors influencing biomass accumulation of green tide causing Ulva spp. on Pyropia culture rafts in the Yellow Sea, China.
Annually recurrent green-tides in the Yellow Sea have been shown to result from direct disposal into the sea of fouling Ulva from Pyropia aquaculture. The role abiotic factors play in Ulva biomass accumulation on rafts was studied to find ways to mitigate this problem. Dissolved inorganic nitrogen (DIN) was very high at all sites, but the highest Ulva biomass was associated with the lowest DIN and anthropogenic N. Under luxuriant background nutrient conditions, variability in temperature and periods of emersion, rather than pH, light and salinity determined Ulva biomass. Two dominant species of Ulva displayed differing tolerances to temperature and desiccation which helped explain why Ulva prolifera dominates floating green-tides. Rather than trying to mitigate green-tides only by reducing nutrient pollution, an earlier harvest of Pyropia in southern Jiangsu Province especially before temperatures increase greatly above 10°C during April, could reduce the biomass of U. prolifera disposed from rafts.


CYT-Rx20 Inhibits Cervical Cancer Cell Growth and Migration Through Oxidative Stress-Induced DNA Damage, Cell Apoptosis, and Epithelial-to-Mesenchymal Transition Inhibition.
The β-nitrostyrene family has been reported to possess anticancer properties. However, the anticancer activity of β-nitrostyrenes on cervical cancer cells and the underlying mechanisms involved remain unexplored. In this study, a β-nitrostyrene derivative CYT-Rx20 (3'-hydroxy-4'-methoxy-β-methyl-β-nitrostyrene) was synthesized, and its anticancer activity on cervical cancer cells and the mechanisms involved were investigated. The effect of CYT-Rx20 on human cervical cancer cell growth was evaluated using cell viability assay. Reactive oxygen species (ROS) generation and annexin V staining were detected by flow cytometry. The protein expression levels of cleaved caspase-3, cleaved caspase-9, cleaved poly (ADPribose) polymerase, γH2AX, β-catenin, Vimentin, and Twist were measured by Western blotting. DNA double-strand breaks were determined by γ-H2AX foci formation and neutral comet assay. Migration assay was used to determine cancer cell migration. Nude mice xenograft was used to investigate the antitumor effects of CYT-Rx20 in vivo. CYT-Rx20 induced cytotoxicity in cervical cancer cells by promoting cell apoptosis via ROS generation and DNA damage. CYT-Rx20-induced cell apoptosis, ROS generation, and DNA damage were reversed by thiol antioxidants. In addition, CYT-Rx20 inhibited cervical cancer cell migration by regulating the expression of epithelial-to-mesenchymal transition markers. In nude mice, CYT-Rx20 inhibited cervical tumor growth accompanied by increased expression of DNA damage marker γH2AX and decreased expression of mesenchymal markers β-catenin and Twist. CYT-Rx20 inhibits cervical cancer cells in vitro and in vivo and has the potential to be further developed into an anti-cervical cancer drug clinically.


HTRF: a technology tailored for biomarker determination-novel analytical detection system suitable for detection of specific autoimmune antibodies as biomarkers in nanogram level in different body fluids.
Classical methods of gene product analysis such as binding assays (e.g., ELISA, protein chip technology) are generally time-consuming, lab-intensive, less sensitive, and lack high-throughput capacity. In addition, all existing methods used to measure proteins necessitate multiple divisions of the original sample and individual tests carried out for each substance, with an associated cost for each test. Together with a small biotech company, we developed a new and innovative analytical detection system based on homogenous time-resolved fluorescence (HTRF) technology. Our system facilitates the development of immune assays that measure selective different analytes such as selected biomarkers in a small sample volume at less than 20 min with a much higher sensitivity compared to common binding assay systems such as enzyme-linked immunosorbent assay (ELISA). Recent advances of the application of this novel detection system combine the power of miniaturization, microfluidics, better linear range, and faster quantification. The power of the HTRF technology offers great promise for point-of-care clinical testing and monitoring of many important analytes such as disease-specific biomarkers in the nanogram level in different human body fluids such as CSF, blood, serum, plasma, and saliva. The linear dynamical range of our HTRF assay was determined between 2.5 and 100 ng/mL. Precision and accuracy calculated for inter- as well as intra-assays was less than ± 10 %. Intra-assay and inter-assay precision for high, medium, and low analyte concentrations show mean CV values less than ± 10 %. Intra- and inter-assay accuracy for all three concentrations show mean recovery values of 80-120 %. The aim of this work is to describe the development and establishment of this novel HTRF system that allows the very fast detection and quantification of biomarkers in different human body fluids. Furthermore, a specific antibody combination that assures a specific binding of the correct refolded autoimmune IgG is evaluated.


Parathyroid hormone early percent change: an individualized approach to predict postthyroidectomy hypocalcemia.
The purpose of this study is to evaluate a percent change model of postoperative parathyroid hormone level in thyroidectomy patients as a predictor of hypocalcemia. Chart review was completed on patients who had undergone total or completion thyroidectomy over a 22-month period in our department. Only those patients with a preoperative ionized calcium and parathyroid hormone (PTH) level and at least 1 postoperative result were included. Ionized calcium levels served as an internal control. The Student t test was used to compare PTH level between the normocalcemic and hypocalcemic groups at each time point. Logistic regression analysis was used to predict hypocalcemia based on the diagnostic criteria. Receiver operator curves were used to maximize sensitivity. Fifty-two patients met the inclusion criteria during the study period. A total of 22 patients (42%) experienced hypocalcemia. We were unable to maximize both sensitivity and specificity at the same time point. When comparing preoperative to 6-hour postoperative PTH percent change, patients with a greater than 44% decrease are likely to have hypocalcemia, with a sensitivity of 100%. Likewise, in those patients without a greater than 44% decrease at 6 hours, early discharge can be considered safe, given the negative predictive value of 100%. In our series, patients with a greater than 44% PTH decrease from preoperative to 6-hour postoperative are very likely to develop hypocalcemia. We would propose that these patients need further inpatient monitoring to progress to safe discharge. Likewise, patients with a less than 44% decrease at the 6-hour time point are unlikely to develop hypocalcemia and may be considered safe for discharge.


Comparative transcriptome analysis identified important genes and regulatory pathways for flower color variation in Paphiopedilum hirsutissimum.
Paphiopedilum hirsutissimum is a member of Orchidaceae family that is famous for its ornamental value around the globe, it is vulnerable due to over-exploitation and was listed in Appendix I of the Convention on International Trade in Endangered Species of Wild Fauna and Flora, which prevents its trade across borders. Variation in flower color that gives rise to different flower patterns is a major trait contributing to its high ornamental value. However, the molecular mechanism underlying color formation in P. hirsutissimum still remains unexplored. In the present study, we exploited natural variation in petal and labellum color of Paphiopedilum plants and used comparative transcriptome analysis as well as pigment measurements to explore the important genes, metabolites and regulatory pathways linked to flower color variation in P. hirsutissimum. We observed that reduced anthocyanin and flavonoid contents along with slightly higher carotenoids are responsible for albino flower phenotype. Comparative transcriptome analysis identified 3287 differentially expressed genes (DEGs) among normal and albino labellum, and 3634 DEGs between normal and albino petals. Two genes encoding for flavanone 3-hydroxylase (F3H) and one gene encoding for chalcone synthase (CHS) were strongly downregulated in albino labellum and petals compared to normal flowers. As both F3H and CHS catalyze essentially important steps in anthocyanin biosynthesis pathway, downregulation of these genes is probably leading to albino flower phenotype via down-accumulation of anthocyanins. However, we observed the downregulation of major carotenoid biosynthesis genes including VDE, NCED and ABA2 which was inconsistent with the increased carotenoid accumulation in albino flowers, suggesting that carotenoid accumulation was probably controlled at post-transcriptional or translational level. In addition, we identified several key transcription factors (MYB73, MYB61, bHLH14, bHLH106, MADS-SOC1, AP2/ERF1, ERF26 and ERF87) that may regulate structural genes involved in flower color formation in P. hirsutissimum. Importantly, over-expression of some of these candidate TFs increased anthocyanin accumulation in tobacco leaves which provided important evidence for the role of these TFs in flower color formation probably via regulating key structural genes of the anthocyanin pathway. The genes identified here could be potential targets for breeding P. hirsutissimum with different flower color patterns by manipulating the anthocyanin and carotenoid biosynthesis pathways.


Critical care in pregnancy.
Childbirth is a major event in the lives of mothers and their families. Critical illness in pregnancy is uncommon but may arise from conditions unique to pregnancy, conditions exacerbated by pregnancy and coincidental conditions. According to the latest Confidential Enquiry into Maternal Deaths in the UK, haemorrhage remains a leading direct cause of mortality; however, there has been an increase in mortality due to indirect causes. The obstetric population has changed over the past decade and we are caring for much older mothers with pre-existing disorders and advanced chronic medical conditions. It is therefore essential to adopt an early multidisciplinary approach for the care of these women. With birth rates increasing, complex caseloads and changes in training of both medical and midwifery staff, the challenge of caring for critically ill obstetric patients requires urgent attention.


Functional identification of fatty acyl reductases in female pheromone gland and tarsi of the corn earworm, Helicoverpa zea.
Most moths utilize sex pheromones released by the female to attract a mate. Females produce the sex pheromone in the pheromone gland in a biosynthetic pathway which consists of several key enzymes. Fatty acyl-CoA reductase is one of the key enzymes, which catalyzes the conversion of fatty acyl-CoA to the corresponding alcohol, playing an important role in producing the final proportion of each pheromone component. In Helicoverpa zea, (Z)-11-hexadecenal is the major sex pheromone component in female pheromone glands and previously a large amount of hexadecanal was also found in female and male tarsi. In our previous study, we compared the transcriptome between pheromone glands and tarsi and found 20 fatty acyl-CoA reductases in both tissues. In this study, we functionally characterized four FARs which were expressed at high levels according to the transcriptome of pheromone glands and tarsi. Fatty acyl-CoA reductase 1 was homologous to other moth pheromone gland specific fatty acyl-CoA reductases, and it was also present in male tarsi. Functional expression in yeast cells indicates that only fatty acyl-CoA reductase 1 was able to produce fatty alcohols. In addition, a decreased mRNA level of fatty acyl-CoA reductase 1 in female pheromone glands and male tarsi by RNAi knockdown caused a significant decrease in the production of (Z)-11-hexadecenal in pheromone glands and hexadecanal in male tarsi. This study is the first to demonstrate the direct function of a fatty acyl-CoA reductase in male tarsi and also confirms its role in sex pheromone biosynthesis in H. zea.


The properties of brain galactocerebroside monolayers.
Using a Langmuir film balance we have compared the properties of films of the brain galactocerebrosides at 37 degrees C. There are two types of cerebroside in brain, those with an alpha-hydroxy substituent on the acyl chain (HFA) and those without (NFA). At equivalent pressures the areas of both cerebroside films are significantly less than the areas of films of the brain glycerolipids, the choline and ethanolamine phosphatides. The isotherm of NFA galactocerebrosides has two discontinuities, one at low and one at high film pressure, while the isotherm of HFA galactocerebrosides is a smooth curve at all film pressures. Below the high-pressure transition the area of the NFA film is significantly larger than the area of the HFA film. When compressed beyond the high-pressure transition there is a marked hysteresis between compression and expansion isotherms of the NFA galactocerebrosides. The pressures of both films continue to rise steeply when they are compressed into areas which are too small for them to exist as simple monolayers. We conclude that under compression cerebroside films form bilayer structures; that bilayer formation starts at low pressure and occurs progressively as the HFA cerebroside monolayer is compressed, but occurs more abruptly in the NFA cerebroside monolayer at the high-pressure-transition region of the isotherm. A study of pure cerebrosides with a single defined acyl chain shows that there is a correlation between the relative volumes of the hydrophobic and hydrophilic parts of the molecule and the ease of bilayer formation. The larger the relative volume of the hydrophilic group the more readily the cerebroside forms a bilayer film. Other brain lipids added to cerebroside monolayers have sharply differing effects on their areas. The areas of films containing cholesterol are less than the areas calculated by adding the areas of the pure components multiplied by their mole fractions. On the other hand, the area of phosphatidylcholine-containing films is much larger than calculated.


Radiosensitivity of different human tumor lines grown as xenografts determined from growth delay and survival data.
Four human tumor lines were grown as xenografts in nude mice to determine whether xenografts derived from different types of tumors would show tumor-type dependent differences in response to single-dose irradiation, and whether these differences, paralleled clinical behavior. Xenografts from a neuroblastoma, a squamous cell carcinoma, a melanoma and a lung adenocarcinoma were studied in terms of growth delay and tumor control dose (TCD50). To exclude an immunoreaction of the host in the radiation response of the tumor xenografts, the tumor lines were tested for their growth in immunosuppressed Wistar rats. No differences in growth of xenografts in either immunodeficient mice or immunosuppressed rats were observed. Both growth delay and local tumor control as expressed by cure correlated well with clinical behavior of the tumor types of origin. This study demonstrates that radiosensitivity of different human tumor lines can be evaluated in terms of growth delay and tumor control dose50 when they are grown as xenografts. To exclude immune reactions, proper controls should be included. The sensitivities established from these evaluations parallel clinical behavior, thus offering a tool for analysis of human tumor radiosensitivity of histologically different tumor types.


Groundwater fluoride and dental fluorosis in southwestern Nigeria.
This study was carried out to assess the fluoride levels of groundwater from open wells, consumed by the residents of three communities located in two distinct geological terrains of southwestern Nigeria. Fluoride concentration was determined using spectrophotometric technique, while analysis of other parameters like temperature, pH and total dissolve solids followed standard methods. Results of the analysis indicated that groundwater samples from Abeokuta Metropolis (i.e., basement complex terrain) had fluoride content in the range of 0.65 ± 0.21 and 1.20 ± 0.14. These values were found to be lower than the fluoride contents in the groundwater samples from Ewekoro peri-urban and Lagos metropolis where the values ranged between 1.10 ± 0.14-1.45 ± 0.07 and 0.15 ± 0.07-2.20 ± 1.41 mg/l, respectively. The fluoride contents in almost all locations were generally higher than the WHO recommended 0.6 mg/l. Analysis of Duncan multiple range test indicated that there is similarity in the level of significance of fluoride contents between different locations of same geological terrain at p ≤ 0.05. It was also observed that fluoride distribution of groundwater samples from the different geological terrain was more dependent on factors like pH and TDS than on temperature. The result of the analyzed social demographic characteristics of the residents indicated that the adults (between the age of 20 and >40 years) showed dental decay than the adolescent (<20 years). This signifies incidence of dental fluorosis by the high fluoride content in the drinking water of the populace. Further investigation on all sources of drinking water and other causes of tooth decay in the area is suggested.


Renal squamous cell carcinoma mimicking xanthogranulomatous pyelonephritis: Case report and review of literature.
Primary renal squamous cell carcinoma (SCC) is a rare primary malignancy of the kidney. Diagnosis is usually delayed because of its lack of characteristic clinical and imaging features and inherent aggressive nature. We present a case of primary renal SCC in a 66-year-old woman with bilateral renal calculi and a complex right lower pole renal mass. The diagnosis of primary renal SCC was established based on the histopathology after right nephrectomy.


Investigations of renal artery blood flow velocity in preterm and term neonates by pulsed Doppler ultrasonography.
So far, echo-Doppler studies of the renal arteries have only been performed in adult subjects. They dealt with qualitative parameters of blood flow, but absolute velocity values could not be determined. This paper presents a method for measurement of absolute values of renal artery blood flow velocity in infancy with a well-defined steep angle of insonation of less than 25 degrees by using a range-gated Doppler system with Duplex-scan. The renal blood flow patterns were studied at specified sites by placement of the transducer below the costal arches on the right flank and left flank, respectively. Thus, a reference collective of 65 infants (26-40 weeks of gestational age, 650-3990 g body weight and 1-145 days of postnatal age at the time of examination) were studied. The time mean of maximum velocity significantly increased with the body weight from 17 cm/s below 1000 g to 29 cm/s above 3000 g of body weight. Whereas the pulsatility indices were independent of body weight. The reproducibility of the method, coefficient of variation (CV) 3%-9%, is comparable with the studies of cerebral vessels.


Tocainide and mortality after myocardial infarction: a prospective study in conscious dogs.
The production of an anterior myocardial infarction as part of an experimental animal model for sudden death was burdened by a persistently elevated mortality rate (30%) despite the use of traditional antiarrhythmic drugs. Mortality was reduced when tocainide (600 mg three times daily) was empirically administered for 4 days before and 4 days after myocardial infarction. Retrospective analysis showed that mortality at 4 days after infarction was significantly lower in the tocainide-treated dogs than in the preceding large group of dogs that had not been so treated (5 [9%] of 55, versus 64 [32%] of 199, p less than 0.01). This difference was still evident 30 days after myocardial infarction (13 [24%] of 55 versus 83 [42%] of 199; p less than 0.05). This observation led to the present prospective study in 106 dogs with a similar protocol but with a randomized sequence. At 4 days after myocardial infarction, the mortality rate was 55% lower in the tocainide group than in the control group (7 [12.5%] of 56 versus 14 [28%] of 50; p less than 0.05). During the 10 days after treatment withdrawal 9 (18%) of 49 dogs in the tocainide group died compared with only 2 (5%) of 36 in the control group. This rebound in mortality produced similar survival figures in the two groups at 14 and at 30 days after infarction when mortality was 30% (17 of 56) in the tocainide group and 34% (17 of 50) in the control group. The tocainide plasma levels were 7.4 +/- 4 micrograms/ml the day before infarction and 7.2 +/- 3 micrograms/ml 3 days after infarction.(ABSTRACT TRUNCATED AT 250 WORDS)


Production of high specific activity 123I for protein iodination for medical use.
Iodine-123 is produced via xenon-133 by irradiation of a sodium iodide target with 108 MeV deuterons from the synchrocyclotron of IPN. The on-line production method is described. The specific activity of the iodine is determined by neutron activation analysis and by a radioimmunological method. The conditions labelling different proteins (insulin, angiotensin) are given and also the purification method to obtain a product ready for injection to patients.


In situ biaxial loading and multi-scale deformation measurements of nanostructured materials at the CoSAXS beamline at MAX IV Laboratory.
Characterization of the mechanical response of polymers and composite materials relies heavily on the macroscopic stress-strain response in uniaxial tensile configurations. To provide representative information, the deformation process must be homogeneous within the gauge length, which is a condition that is rarely achieved due to stress concentration or inhomogeneities within the specimen. In this work, the development of a biaxial mechanical testing device at the CoSAXS beamline at MAX IV Laboratory is presented. The design facilitates simultaneous measurement of small- and wide-angle X-ray scattering (SAXS/WAXS), allowing assessment of the microstructural configuration before, after and during the continuous deformation process at multiple length scales. The construction also supports multiple deformation conditions, while guaranteeing stability even at high loads. Furthermore, the mechanical experiments can be complemented with spatially resolved mesoscopic surface deformation measurements using 3D-surface digital image correlation (DIC). Polycarbonate (PC) was used to demonstrate the varied material response to multi-axial deformation, as PC is isotropic with a high glass transition temperature (∼150°) and high strength. As a result, a clear correlation between full-field methods and the microstructural information determined from WAXS measurements is demonstrated. When a uniaxial load is applied, homogeneous strain regions could be observed extending perpendicular to the applied load. When a secondary axial load was added (biaxial mode), it was observed that high strain domains were created near the centre of the sample and at the boundaries after yield. With increased strain, the deformation in the main deformation direction also increases. Mechanical reliability was demonstrated by carrying out static loading of polyacrylonitrile-based carbon fibre (CF) bundles. As a result, the nonlinear stiffening behaviour typically observed in CFs was seen, while no evidence of the creation of new voids during loading was observed. The results support the reliability and broad applicability of the developed technique.


Effect of natural mediators on the stability of Trametes trogii laccase during the decolourization of textile wastewaters.
The purpose of the present study was to determine the effect of natural mediators on the stability of the Trametes trogii crude laccase in the process of decolourization of textile effluents. Acetosyringone allowed the highest wastewaters decolourization rate of 25%. At higher concentrations of acetosyringone, the relative activity of laccase decreased approximately by between 38% and 88% after 5 days of incubation. T. trogii laccase was strongly inactivated at 3 mM syringaldehyde, after 3 days of incubation. However, laccase activity is more stable in the presence of the vanillin and m-coumarate. The T. trogii growth on solid effluent-based-medium was examined and evaluated by measuring the colony diameter in cm. T. trogii was completely inhibited on 100:0 and 80:20 effluent:water solid medium, however, colony diameter reached 5 cm on 60:40 effluent:water solid medium after 13-14 days incubation. When the textile effluent was pre-treated with laccase and laccase-acetosyringone system, the colony diameter of 2 cm of T. trogii on 80:20 effluent:water solid medium was reached after 14 and 10 days of incubation respectively.


The effect of amitriptyline on the central nervous system penetration of methotrexate.
The reported ability of amitriptyline to enhance the penetration of diffusion limited substances across the blood brain barrier was investigated. The CSF:plasma ratio of methotrexate in primates was not significantly altered by amitriptyline pretreatment. Nor was methotrexate plasma clearance altered.


A case series of five infants with scimitar syndrome.
Scimitar syndrome is a rare association of congenital cardiopulmonary anomalies consisting of a partial anomalous pulmonary venous connection of the right lung to the inferior vena cava, right lung hypoplasia, dextroposition of the heart, and anomalous systemic arterial supply to the right lung. To date, this syndrome has not been reported or discussed in the nursing literature. In this article, 5 patients are presented to illustrate the variable presentation of this syndrome, ranging from infants who are asymptomatic to those with heart failure and severe pulmonary hypertension. The clinical signs and symptoms, diagnostic dilemmas, and medical and surgical management are discussed. The triad of respiratory distress, right lung hypoplasia, and dextroposition of the heart should alert the clinician to the possibility of this syndrome.


Hypereosinophilic syndrome with cardiac involvement: early diagnosis by cardiac magnetic resonance imaging.
Cardiac manifestation is the major cause of morbidity in patients with hypereosinophilic syndrome (HES) if left untreated. Endomyocardial biopsy is currently the gold standard for diagnosis, although typical findings using noninvasive methods such as cardiac magnetic resonance (CMR), may suggest the diagnosis. The present case provides an example of the beneficial use of cardiac magnetic resonance for early diagnosis and follow-up of patients with cardiac involvement in hypereosinophilic syndrome.


Latent profile analysis of diurnal cortisol patterns at the ages of 2, 3.5, and 5 years: Associations with childcare setting, child individual characteristics, and maternal distress.
This study performed latent profile analysis from more than 4000 saliva cortisol samples collected from children at the ages of 2 (T1), 3.5 (T2), and 5 years (T3). Three clearly different cortisol profiles were identified. The largest group at every age point was the Low/Regular latent profile, in which the cortisol slopes followed typical diurnal variation. A smaller proportion of the children belonged to the latent profile with relatively Low/Flat slope, and a minority belonged to the High/Fluctuating latent group, where the overall cortisol values and variations between the slopes were clearly higher than in the other groups. Most of the children who belonged to the High/Fluctuating group were cared for at home, they had higher temperamental surgency and their mothers had more depressive symptoms than in the other latent profile groups. However, only moderate intraindividual stability in diurnal cortisol profiles was observed across the follow-up period. On average, half of the children moved between the groups from T1 to T3. Neither child temperament, social competence, nor sex explained the stability or movement between the groups across age. Variations in cortisol profiles may be caused by the child's age, and diurnal cortisol rhythm becomes more regular along with development. Methodological issues regarding saliva cortisol research in young children are discussed. Also, more longitudinal research is needed to clarify mechanisms between environmental as well as individual factors and possible dysregulation in a child's HPA axis functioning.


Rate of perfusion modulates colloidal carbon leakage from rat intestinal microvessels in vitro.
In order to investigate the effects of varying the rate of flow on endothelial integrity the rat isolated small intestinal vasculature was perfused at 1, 5, 10 or 20 ml/min with a gelatin-containing physiological salt solution (GPSS), followed by an injection of colloidal carbon suspension (CC). Significantly greater microvascular CC leakage occurred at 1 or 5 ml/min than at 10 or 20 ml/ mitt. CC leakage at the two slower rates of flow was reduced by adding red blood cells to the GPSS, suggesting that the microvascular endothelium became hypoxic when perfused with GPSS at 1 or 5 ml/min. After perfusion at 20 ml/min with GPSS containing resiniferatoxin (1 muM) or 5-hydroxytryptamine (100 muM), CC leakage was significantly lower than after similar perfusion at 10 ml/min. Two nitric oxide (NO) synthesis blockers, N-nitro-L-arginine methyl ester (L-NAME, 100 muM) and methylene blue (20 muM), and an NO scavenger CPTIO (100 muM) each increased CC leakage. This suggests that NO was being produced at perfusion rates of 10 or 20 ml/min. Sodium nitroprusside (10 muM), 8-bromo-cGMP (100 muM) and BN52021 (10 muM) each significantly reduced CC leakage in the presence of L-NAME.


Effect of morning bright light treatment for rest-activity disruption in institutionalized patients with severe Alzheimer's disease.
Disturbances in rest-activity rhythm are prominent and disabling symptoms in Alzheimer's disease (AD). Nighttime sleep is severely fragmented and daytime activity is disrupted by multiple napping episodes. In most institutional environments, light levels are very low and may not be sufficient to enable the circadian clock to entrain to the 24-hour day. The purpose of this randomized, placebo-controlled, clinical trial was to test the effectiveness of morning bright light therapy in reducing rest-activity (circadian) disruption in institutionalized patients with severe AD. Subjects (n = 46, mean age 84 years) meeting the NINCDS-ADRDA (National Institute of Neurological and Communicative Disorders and Stroke--the Alzheimer's Disease and Related Disorders Association) AD diagnostic criteria were recruited from two large, skilled nursing facilities in San Francisco, California. The experimental group received one hour (09:30-10:30) of bright light exposure (> or = 2500 lux in gaze direction) Monday through Friday for 10 weeks. The control group received usual indoor light (150-200 lux). Nighttime sleep efficiency, sleep time, wake time and number of awakenings and daytime wake time were assessed using actigraphy. Circadian rhythm parameters were also determined from the actigraphic data using cosinor analysis and nonparametric techniques. Repeated measures analysis of variance (ANOVA) was used to test the primary study hypotheses. Although significant improvements were found in subjects with aberrant timing of their rest-activity rhythm, morning bright light exposure did not induce an overall improvement in measures of sleep or the rest-activity in all treated as compared to control subjects. The results indicate that only subjects with the most impaired rest-activity rhythm respond significantly and positively to a brief (one hour) light intervention.


Correlation between fracture toughness and leucite content in dental porcelains.
To determine the correlation between fracture toughness and leucite content in dental porcelains. The mechanisms by which leucite influences the fracture toughness of dental porcelains were also investigated. Six porcelains were tested: A (Ceramco I/Dentsply), B (Ceramco II/Dentsitply), C (Finesse/Dentsply), D (d.Sign/Ivoclar), Cb (Cerabien/Noritake) and V (Vitadur Alpha/Vita). Bar-shaped specimens were produced, and their fracture toughness was determined by means of the single-edge precracked beam (SEPB) method. The test consisted of fracturing the specimen after a precrack was generated by a bridge-anvil device. KIc was calculated based on fracture force and size of the precrack. Microstructural analysis and determination of the leucite volume fraction were performed on polished specimens etched with 2% HF for 15s by means of scanning electron microscopy. Fractographic analysis was performed on fracture surfaces. Porcelains A and B presented the highest leucite contents (22%) and similar KIc values (1.23 and 1.22 MPa m1/2, respectively), significantly higher than the other materials. Porcelains C and D presented similar K(Ic) values (0.81 and 0.93 MPa m1/2, respectively), but different leucite contents (6 and 15%, respectively). Porcelain D presented higher KIc compared to porcelains Cb and V (0.71 and 0.75 MPa m1/2, respectively), which presented similar values and the lowest leucite contents (0%). Fractographic analysis showed that porcelains with higher leucite content presented higher incidence of crack deflection. For the materials evaluated in this study, the leucite content was directly related to KIc. The main toughening mechanism observed was crack deflection around leucite particles and clusters.


Cancer surveillance for transgender and gender diverse patients with Lynch syndrome: a practice resource of the Collaborative Group of the Americas on Inherited Gastrointestinal Cancer.
Transgender and gender diverse (TGD) populations with hereditary cancer syndromes face unique obstacles to identifying and obtaining appropriate cancer surveillance and risk-reducing procedures. There is a lack of care provider knowledge about TGD health management. Lynch syndrome (LS) is one of the most common hereditary cancer syndromes, affecting an estimated 1 in 279 individuals. There are no clinical guidelines specific for TGD individuals with LS, highlighting a need to improve the quality of care for this population. There is an urgent need for cancer surveillance recommendations for TGD patients. This commentary provides recommendations for cancer surveillance, risk-reducing strategies, and genetic counseling considerations for TGD patients with LS.


Intrinsic antiviral immunity of barrier cells revealed by an iPSC-derived blood-brain barrier cellular model.
Physiological blood-tissue barriers play a critical role in separating the circulation from immune-privileged sites and denying access to blood-borne viruses. The mechanism of virus restriction by these barriers is poorly understood. We utilize induced pluripotent stem cell (iPSC)-derived human brain microvascular endothelial cells (iBMECs) to study virus-blood-brain barrier (BBB) interactions. These iPSC-derived cells faithfully recapitulate a striking difference in in vivo neuroinvasion by two alphavirus isolates and are selectively permissive to neurotropic flaviviruses. A model of cocultured iBMECs and astrocytes exhibits high transendothelial electrical resistance and blocks non-neurotropic flaviviruses from getting across the barrier. We find that iBMECs constitutively express an interferon-induced gene, IFITM1, which preferentially restricts the replication of non-neurotropic flaviviruses. Barrier cells from blood-testis and blood-retinal barriers also constitutively express IFITMs that contribute to the viral resistance. Our application of a renewable human iPSC-based model for studying virus-BBB interactions reveals that intrinsic immunity at the barriers contributes to virus exclusion.


Characterization of a group unrelated patients with arthrogryposis multiplex congenita.
Arthrogryposis multiplex congenita is a relatively rare neuromuscular syndrome, with a prevalence of 1:3000-5000 newborns. In this study, the authors describe the clinical features of a group of 50 unrelated Mexican patients with arthrogryposis multiplex congenita. Patients were diagnosed by physical and radiographic examination and the family history was evaluated. Of the 50 cases, nine presented other features (pectum excavatum, cleft palate, mental retardation, ulnar agenesis, etc.). Environmental factors, as well as prenatal and family history, were analyzed. The chromosomal anomalies and clinical entities associated with arthrogryposis multiplex congenita were reported. No chromosomal aberrations were present in the cases with mental retardation. Three unrelated familial cases with arthrogryposis multiplex congenita were observed in which autosomal recessive, autosomal dominant and X-linked inheritance patterns are possible. A literature review regarding arthrogryposis multiplex congenita was also conducted. It is important to establish patient-specific physical therapy and rehabilitation programs. A multidisciplinary approach is necessary, with medical, surgical, rehabilitation, social and psychological care, including genetic counseling.


Cardiac macrophages adopt profibrotic/M2 phenotype in infarcted hearts: Role of urokinase plasminogen activator.
Macrophages (mac) that over-express urokinase plasminogen activator (uPA) adopt a profibrotic M2 phenotype in the heart in association with cardiac fibrosis. We tested the hypothesis that cardiac macs are M2 polarized in infarcted mouse and human hearts and that polarization is dependent on mac-derived uPA. Studies were performed using uninjured (UI) or infarcted (MI) hearts of uPA overexpressing (SR-uPA), uPA null, or nontransgenic littermate (Ntg) mice. At 7days post-infarction, cardiac mac were isolated, RNA extracted and M2 markers Arg1, YM1, and Fizz1 measured with qrtPCR. Histologic analysis for cardiac fibrosis, mac and myofibroblasts was performed at the same time-point. Cardiac macs were also isolated from Ntg hearts and RNA collected after primary isolation or culture with vehicle, IL-4 or plasmin and M2 marker expression measured. Cardiac tissue and blood was collected from humans with ischemic heart disease. Expression of M2 marker CD206 and M1 marker TNFalpha was measured. Macs from WT mice had increased expression of Arg1 and Ym1 following MI (41.3±6.5 and 70.3±36, fold change vs UI, n=8, P<0.007). There was significant up-regulation of cardiac mac Arg1 and YM1 with MI in both WT and uPA null mice (n=4-9 per genotype and condition). Treatment with plasmin increased expression of Arg1 and YM1 in cultured cardiac macs. Histologic analysis revealed increased density of activated fibroblasts and M2 macs in SR-uPA hearts post-infarction with associated increases in fibrosis. Cardiac macs isolated from human hearts with ischemic heart disease expressed increased levels of the M2 marker CD206 in comparison to blood-derived macs (4.9±1.3). Cardiac macs in mouse and human hearts adopt a M2 phenotype in association with fibrosis. Plasmin can induce an M2 phenotype in cardiac macs. However, M2 activation can occur in the heart in vivo in the absence of uPA indicating that alternative pathways to activate plasmin are present in the heart. Excess uPA promotes increased fibroblast density potentially via potentiating fibroblast migration or proliferation. Altering macrophage phenotype in the heart is a potential target to modify cardiac fibrosis.


Genital Sparing Robot-Assisted Radical Cystectomy with Intracorporeal Neobladder & Paravaginal Repair.
Vaginal prolapse is a known complication after radical cystectomy, requiring additional procedures in 10% of the patients.1 This results from loss of level I and II vaginal support due to the removal of pelvic structures. In addition, a neobladder urinary diversion, with Valsalva voiding, predisposes to vaginal prolapse. A genital-sparing approach with paravaginal repair can help prevent such complications. The genital sparing technique preserves the uterus, fallopian tubes, ovaries, and vagina, while paravaginal repair involves suturing of the lateral vaginal wall to the arcuate fascia located on the medial aspect of the obturator internus muscle. The procedure begins by placing the patient in a lithotomy position, with a steep Trendelenburg. Standard 6 port cystectomy configuration is utilized with an additional 15 mm port for bowel anastomosis. Initially, the ureters and lateral bladder space are mobilized. Posteriorly a dissection plane is developed separating the bladder from the anterior vaginal wall. Distal dissection is carefully performed in that plane to avoid disrupting the urethral-external sphincter complex. Then the bladder is dropped from anterior attachments, the Dorsal venous complex (DVC) and bladder neck are exposed. Urethra is transected distal to the bladder neck, after circumferential mobilization, to complete the cystectomy, again avoiding disruption of the continence mechanism, and opening the endo-pelvic fascia. Cystectomy and pelvic lymph node dissection are completed in a standard fashion. The arcuate fascia is identified bilaterally for level I paravaginal repair. The lateral aspect of the paravaginal tissue is secured to this ligament, using 3 interrupted Polydioxanone (PDS) sutures, bilaterally. An ileal "Hautman's W pouch" neobladder is constructed using 50 cm of the small intestine, similar to the previously reported technique.2 Bricker-type uretero-ileal anastomosis is performed over a double J stent. Bowel continuity is restored by a side-to-side anastomosis using endo-GIA (gastrointestinal anastamosis EndoGIATM ) staplers. No intra or postoperative complications were noted. Robot dock time was 8 hours and 23 minutes with an EBL of 100 mL. The patient was discharged on post operative day (POD) 6 and Foley catheter with ureteral stents was removed on POD 27 after a cystogram confirmed no leaks. At 6-month follow-up, the patient reported good continence using a single pad, voiding every 3-4 hours. Fluoro-urodynamics demonstrated 651 mL capacity, low-pressure voiding, minimal residual urine, and no reflux. No prolapse was noted on fluoroscopy and pelvic examination with the Valsalva maneuver. The patient reported a good satisfaction level, regarding her urinary symptoms. We report satisfactory short-term outcomes of a feasible technique to prevent postcystectomy prolapse; however, long-term follow-up of a larger cohort can help establish its efficacy.


Reliability of Fractional Flow Reserve and Instantaneous Wave-Free Ratio in Assessing Intermediate Coronary Stenosis in Patients With Atrial Fibrillation.
Despite the current use of fractional flow reserve (FFR) and instantaneous wave-free ratio (iFR) for guiding revascularization in atrial fibrillation (AF), there is a lack of studies evaluating their reliability in this particular population. This retrospective study aimed to investigate the reliability of FFR and iFR in patients with AF. This retrospective study included all patients with AF undergoing FFR measurements (n = 45 vessels from 36 patients) at Brugmann University Hospital, Brussels, Belgium, between 2012 and 2020 or iFR (n = 18 vessels from 13 patients) and a corresponding number of patients with sinus rhythm (SR) randomly selected from the same period, benefiting from iFR (n = 20 vessels from 17 patients) or FFR (n = 50 vessels from 37 patients). Our main findings indicate that there is an increased beat-to-beat variability of individual iFR measures in patients with AF, compared with SR. In addition, the reproducibility of iFR on test-retest is low in patients with AF, leading to increased lesion reclassification (53.8% of lesions reclassified on 2 consecutive iFR measurements in AF vs 6.6% lesions reclassified in SR, p <0.05). In contrast, FFR seems to be more robust in evaluating coronary lesions in AF in terms of equivalent variability, reproducibility, and lesion reclassification observed in the SR population. In conclusion, this is the first study to evaluate the reliability of iFR and FFR in AF. Our findings raise caution in using iFR to guide revascularization in patients with AF, whereas FFR seems to be more robust in this population.


DNA methylation of hypertension-related genes and effect of riboflavin supplementation in adults stratified by genotype for the MTHFR C677T polymorphism.
The interaction between genetic, epigenetic and environmental factors plays an important role in the aetiology of hypertension. GWAS and observational studies link the C677T polymorphism in methylenetetrahydrofolate reductase (MTHFR) with hypertension, while riboflavin, the MTHFR cofactor, has been shown to reduce blood pressure and global DNA methylation in homozygous (TT genotype) individuals. It is currently unclear whether riboflavin modulates DNA methylation of other hypertension-related genes. To compare DNA methylation of hypertension-related genes in adults stratified by MTHFR genotype and effect of riboflavin intervention in adults with the variant MTHFR 677TT genotype. Pyrosequencing was carried out for hypertension-related genes (ACE, AGTR1, GCK, GNA12, IGF2, MMP9 and NOS3) in blood samples from participants in previous trials (CC, n = 40; TT, n = 40). The effect of intervention with riboflavin (1.6 mg/d for16 weeks) or placebo on DNA methylation was investigated in adults with the variant MTHFR 677TT genotype (n = 80). Individuals with the MTHFR 677TT v CC genotype had significantly higher average DNA methylation at NOS3 (+1.66%, P = 0.044). In response to riboflavin supplementation in TT individuals, there was an increase in average DNA methylation at IGF2 (+1.09%, P = 0.019) and a decrease at ACE (-0.44%, P = 0.021) in females only. Specific CpG sites were hypomethylated in GNA12 and hypermethylated in AGTR1. This study provides the first RCT evidence that riboflavin alters DNA methylation of hypertension-related genes in adults with the MTHFR 677TT genotype, providing some insight into mechanisms linking hypertension with the genotype-specific response of BP to riboflavin.


Small sum privacy and large sum utility in data publishing.
While the study of privacy preserving data publishing has drawn a lot of interest, some recent work has shown that existing mechanisms do not limit all inferences about individuals. This paper is a positive note in response to this finding. We point out that not all inference attacks should be countered, in contrast to all existing works known to us, and based on this we propose a model called SPLU. This model protects sensitive information, by which we refer to answers for aggregate queries with small sums, while queries with large sums are answered with higher accuracy. Using SPLU, we introduce a sanitization algorithm to protect data while maintaining high data utility for queries with large sums. Empirical results show that our method behaves as desired.


Alpha-1-acid glycoprotein concentration and the protein binding of disopyramide in healthy subjects.
Age- and gender-related changes in serum alpha 1-acid glycoprotein (AAG) concentration and the serum protein binding of disopyramide were examined after intensive medical examination. Based on the clinical chemistry tests over 51 points, 245 subjects were diagnosed as healthy and 71 subjects (22.5%) revealed an abnormal value for at least one item. In the healthy subjects, serum AAG concentration in men was significantly higher than in women (men, 0.78 +/- 0.18 mg/mL, mean +/- SD; women, 0.67 +/- 0.16 mg/mL). In contrast, there were no significant differences in the AAG concentration between age groups for men and women and in the unbound fraction of disopyramide. Gender changes AAG concentration. Age, however, does not change AAG concentration and the protein binding of the basic drug.


Nevada gaming: revenues and taxes (1945-95).
In 1931, Nevada legalized casino gambling, an act which allowed its "gaming industry" to develop. Because many jurisdictions outside Nevada are now embracing legalized gambling as a path to a brighter economic future and because this proliferation presents Nevada with new challenges and opportunities, it is a good time to review the Nevada experience. Here, the focus is on gaming revenues and gaming tax collections during the 1945-95 period. We find that the growth rate of Nevada's gaming industry has slowed over time, with the relative importance of gaming in the Nevada economy peaking in the late 1970's. The Nevada economy has since been gradually diversifying, something which will increasingly pressure Nevadans to look for government revenue sources other than gaming if current levels of government services are to be maintained.


The Binding Interface between Human APOBEC3F and HIV-1 Vif Elucidated by Genetic and Computational Approaches.
APOBEC3 family DNA cytosine deaminases provide overlapping defenses against pathogen infections. However, most viruses have elaborate evasion mechanisms such as the HIV-1 Vif protein, which subverts cellular CBF-β and a polyubiquitin ligase complex to neutralize these enzymes. Despite advances in APOBEC3 and Vif biology, a full understanding of this direct host-pathogen conflict has been elusive. We combine virus adaptation and computational studies to interrogate the APOBEC3F-Vif interface and build a robust structural model. A recurring compensatory amino acid substitution from adaptation experiments provided an initial docking constraint, and microsecond molecular dynamic simulations optimized interface contacts. Virus infectivity experiments validated a long-lasting electrostatic interaction between APOBEC3F E289 and HIV-1 Vif R15. Taken together with mutagenesis results, we propose a wobble model to explain how HIV-1 Vif has evolved to bind different APOBEC3 enzymes and, more generally, how pathogens may evolve to escape innate host defenses.


Endoscopic submucosal dissection for anastomotic lesions after colorectal surgery.
Endoscopic submucosal dissection (ESD) for anastomotic lesions is technically challenging. We aimed to characterize the clinicopathologic characteristics, feasibility, and effectiveness of ESD for anastomotic lesions of the lower gastrointestinal tract. We retrospectively investigated 55 patients with anastomotic lesions of the lower gastrointestinal tract who underwent ESD from February 2008 to January 2021. The lesions involving one or both sides of anastomoses were classified into the unilaterally involving anastomosis (UIA) or straddling anastomosis (SA) group, respectively. We collected clinicopathological characteristics, procedure-related parameters and outcomes, and follow-up data and analyzed the impact of anastomotic involvement. The mean age was 62.5 years, and the median procedure duration was 30 min. The rates of en bloc resection and R0 resection were 90.9% and 85.5%, respectively. Four patients (7.3%) experienced major adverse events (AEs). During a median follow-up of 66 months (range 14-169), seven patients had local recurrence, and six patients had metastases. The 5-year disease-free survival and overall survival rates were 82.4% and 90.7%, respectively. The 5-year disease -specific survival (DSS) rate was 93.3%. Compared with the UIA group, the SA group had significantly longer procedure duration, larger specimen, lower rates of en bloc resection and R0 resection, and shorter disease-free survival (all P < 0.05). However, rates of AEs did not differ significantly between the two groups. The short-term and long-term outcomes of ESD for colorectal anastomotic lesions were favorable. Although with technically challenging, ESD could be performed safely and effectively for lesions at the anastomoses.


In vitro cultured Neoparamoeba perurans causes amoebic gill disease in Atlantic salmon and fulfils Koch's postulates.
Amoebic gill disease (AGD) in marine farmed Atlantic salmon is of growing concern worldwide and remains a significant health issue for salmon growers in Australia. Until now the aetiological agent, Neoparamoeba perurans, has not been amenable to in vitro culture and therefore Koch's postulates could not be fulfilled. The inability to culture the amoeba has been a limiting factor in the progression of research into AGD and required the maintenance of an on-going laboratory-based infection to supply infective material. Culture methods using malt yeast agar with sea water overlaid and subculturing every 3-4 days have resulted in the establishment of a clonal culture of N. perurans, designated clone 4. Identity of the amoeba was confirmed by PCR. After 70 days in culture clone 4 infected Atlantic salmon, causing AGD, and was re-isolated from the infected fish. Diagnosis was confirmed by histology and the infectious agent identified by PCR and in situ hybridisation using oligonucleotide primers and probes previously developed and specific to N. perurans. This study has fulfilled Koch's postulates for N. perurans as a causative agent of AGD and illustrates its free-living and parasitic nature.


Oral hygiene behaviour automaticity: Are toothbrushing and interdental cleaning habitual behaviours?
Toothbrushing and interdental cleaning are critical to maintaining good oral health. Literature is beginning to suggest that these behaviours may be conducted automatically, although the instigation ('deciding to do') and execution ('doing') of these behaviours has never been examined separately. The objective of this study was to test a theoretically informed supposition that oral hygiene behaviours in adults are automatic behaviours. One hundred and fifty participants attending three types of dental providers covering emergency and routine dental services, completed a questionnaire. The self-reported behavioural automaticity index scale (SRBAI) was used to measure behavioural automaticity. Morning toothbrushing SRBAI scores were higher than evening scores (Z=-3.315, p = 0.001). Automaticity scores for instigating both toothbrushing and interdental cleaning were also higher compared to execution (toothbrushing: Z=-2.601, p = 0.009 and interdental cleaning: Z=-2.256. p = 0.024). Toothbrushing automaticity scores were associated with age, gender and self-efficacy, whereas interdental cleaning automaticity scores were associated with intention. Individuals in lower socio-economic status (SES) occupations had significantly higher automaticity scores for interdental cleaning compared to those with higher SES roles. A high proportion of people undertake toothbrushing and interdental cleaning automatically, especially in relation to embarking on these behaviours. This is most pronounced in morning toothbrushing. Promoting improvements in oral hygiene behaviour toothbrushing and interdental cleaning are key strategies in preventing caries and periodontal disease. A greater understanding of what prompts and sustains these behaviours helps inform how behaviour change efforts should be approached.


Ecologically relevant genetic variation from a non-Arabidopsis perspective.
Ecologically relevant genetic variation occurs in genes harbouring alleles that are adaptive in some environments but not in others. Analysis of this type of genetic variation in model organisms has made substantial progress, and is now being expanded to other species in order to better cover the diversity of plant life. Recent advances in connecting ecological and molecular studies in non-model species have been made with regard to edaphic and climatic adaptation, plant reproduction, life-history parameters and biotic interactions. New research avenues that increase biological complexity and ecological relevance by integrating ecological experiments with population genetic and functional genomic approaches provide new insights into the genetic basis of ecologically relevant variation.


Advances in Contrast Agents for Contrast-Enhanced Magnetic Resonance Imaging.
Magnetic resonance imaging (MRI) is a well-established medical invention in modern medical technology diagnosis. It is a nondestructive, versatile, and sensitive technique with a high spatial resolution for medical diagnosis. However, MRI has some limitations in differentiating certain tissues, particularly tiny blood vessels, pathological to healthy tissues, specific tumors, and inflammatory conditions such as arthritis, atherosclerosis, and multiple sclerosis. The contrast agent (CA) assisted imaging is the best possible solution to resolve the limitations of MRI. The literature review was carried out using the keywords, "MRI, T1&T2 relaxation, MRI CAs, delivery and adverse effects, classification of CAs." The tools used for the literature search were PubMed, Scopus, and Google Scholar. The literature findings focus on MRI technique, limitations, and possible solutions. Primarily, the review focuses on the mechanism of CAs in image formation with detailed explanations of T1 and T2 relaxations, the mechanism of the MRI-CA image formations. This review presents the adverse effects of CA as well as available marketed formulations and recent patents to extent complete information about the MRI-CA. MRI generates detailed visual information of various tissues with high resolution and contrast. The proton present in the biological fluid plays a crucial role in MR image formation, and it is unable to distinguish pathological conditions in many cases. The CAs are the best solution to resolve the limitation by interacting with native protons. The present review discusses the mechanism of CAs in contrast enhancement and its broad classification with the latest literature. Furthermore, the article presents information about CA biodistribution and adverse effects. The review concludes with an appropriate solution for adverse effects and presents the future prospective for researchers to develop advanced formulations.


The Possibility of Including Donkey Meat and Milk in the Food Chain: A Southern African Scenario.
Animal proteins are essential for the optimal growth and health of humans. Meat and milk are common sources of protein, mostly produced by ruminants. The agrarian challenges experienced around the world warrant sourcing alternative proteins from animals that can withstand harsh environmental conditions to produce quality proteins. Donkeys (Equus asinus) are known to survive on low husbandry and inferior quality forage to produce meat and milk, which have been used since ancient times. However, the commercialisation of these products has not flourished due to product scarcity, low production rates, and consumer preferences. Recent discoveries and the quest to look for alternative sources of protein have sparked studies on donkey products. In addition, milk and meat from donkeys have active ingredients that could also contribute to curing diseases. Donkey milk is believed to contain antioxidant, antimicrobial, antiproliferative, and antidiabetic properties. In many countries, particularly in Africa, the consumption of donkey meat and milk has not been fully adopted due to a lack of knowledge and legislation regarding production.


[The effect of ts-mutation on expression of genes induced by heat shock in Drosophila melanogaster. III. Synthesis of proteins cognate to HSP70].
2D-electrophoresis performed as described by O'-Farrel has revealed clear-cut differences in the pattern of proteins synthesized in the cells of wild-type flies and in the cells of ts-lethal. The cells of the mutant studied after heat-shock exhibit not only the lack of HSP83 and HSP35 but also the absence of a few of the heat-shock proteins belonging to the HSP70 group. Moreover, in the cells of the mutant an intensive synthesis of a protein with mol. weight 72 KDa was observed after heat-shock. This protein belongs to highly abundant heat-shock cognate proteins (HSCP) which are usually not induced by temperature elevation.