Atomoxetine Protects Against NMDA Receptor-mediated Hippocampal Neuronal Death Following Transient Global Cerebral Ischemia.
Atomoxetine has been widely used for the treatment of attention-deficit/ hyperactivity disorder. ATX has additional abilities such as antagonistic effects on the N-methyl-Daspartate receptors (NMDARs) and benefit effects in some animal models of neurological disorders. However, there were few studies regarding protective effects and related mechanisms of ATX against cerebral ischemic insults. The objective of this study is to investigate neuroprotection of ATX pretreatment and its mechanisms in the hippocampal cornu ammonis 1 (CA1) region following transient global cerebral ischemia in gerbils. Gerbils were subjected to transient global cerebral ischemia induced by the occlusion of common carotid arteries for 5 min. Thirty mg/kg of ATX was administrated intraperitoneally once daily for 3 days before ischemic surgery. To examine neuroprotective effects of ATX, we carried out neuronal nuclear antigen immunohistochemistry and Fluoro-Jade B histofluorescence staining. In addition, immunoreactivities of NMDAR1, NMDAR2A/B, B-cell lymphoma 2 (Bcl-2) and Bcl-2- associated X protein (Bax) are closely related with neuroexcitotoxicity. ATX pretreatment reduced ischemia-induced hyperactivity and protected CA1 pyramidal neurons from ischemia. Pretreatment with ATX significantly attenuated ischemia-induced increases of NMDAR1 and NMDAR2A/B immunoreactivities in the CA1 pyramidal neurons at early time following ischemia. In addition, significant ischemia-induced alterations of Bcl-2 and Bax immunoreactivities were not observed in the ATX-treated group following ischemia. These results show that pretreatment with ATX protected against ischemic neuronal via inhibition of ischemia-induced excitotoxicity at early time following transient global cerebral ischemia.


Relationships between immunoglobulin and fat-soluble vitamins in colostrum of Japanese Black multiparous cows.
Data from 19 Japanese Black multiparous cows were collected to clarify the relationships among immunoglobulin (Ig) G, IgA, β-carotene, vitamin A and α-tocopherol contents in colostrum of cows in order to evaluate the role of fat-soluble vitamins on colostral IgG and IgA production. Mean colostral IgG was 141 mg/mL, ranging from 65 to 208 mg/mL, whereas mean colostral IgA was 8.7 mg/mL, ranging from 1.0 to 34.6 mg/mL. Colostral IgG increased with aging in multiparous cows. There were positive correlations between colostral IgG and colostral vitamin A or colostral α-tocopherol in cows, and the higher adjusted R(2) was obtained in the prediction model of colostral IgG from age and colostral vitamin A. Colostral vitamin A was positively correlated with colostral β-carotene or colostral α-tocopherol in cows, but there were no relationships between colostral IgA and colostral IgG or colostral fat-soluble vitamins. These results indicate that fat-soluble vitamin contents in colostrum of cows may change in similar patterns and high colostral vitamin A is related with high colostral IgG.


Luminescence and structure of the protonated forms of meso-tetraarylporphyrins in solution.
A comparative study of the spectral-luminescent properties (at 293 K and 77 K) of meso-tetrakis(o-tolyl)porphyrin (o-TTP) and meso-tetraphenylporphyrin (TPP), and also meso-tetrakis(n-propyl)porphyrin (TPrP) in nonaqueous acid media has been carried out with the aim to reveal effects associated with the influence of methyl groups which hinder sterically the twist of the phenyl rings enhancing their conjugation with the macrocycle in acid medium. On the basis of the spectral data it is concluded that although the introduction of the methyl groups diminishes this twist of phenyls it does not eliminate it completely (the absorption spectra of protonated o-TTP are intermediate between those of TTP and TPrP). The fluorescence spectra of the protonated forms of o-TTP and TPP at 77 K are approximately mirror-symmetrical to the Q(0-0) and Q(0-1) absorption bands; on the elevation of the temperature up to 293 K the fluorescence and absorption bands broaden, the Stokes shift largely increases, and the mirror symmetry of the spectra disappears, which is indicative of changes in solute-solvent interactions in the S1 state. The analysis of the obtained data on the energetics of photophysical processes leads to the conclusion that the significant role of radiationless deexcitation in the S1-->S0 channel, characteristic of the protonated form of TPP, is retained at 77 K. The introduction of the methyl groups at the ortho positions sharply decreases the corresponding rate constant, so that the S1-->T1 intersystem crossing becomes the main channel of deexcitation. The influence of heavy atoms (the Cl- and Br- anions) on the luminescence of the protonated forms has been investigated. It is shown that, analogous to the case of octaethylporphyrin investigated earlier, in nonaqueous media the protonated species of o-TTP form bis adducts of the dihydrochloride type. Contrary to this, the experiments with meso-tetrakis(p-sulfophenyl)porphyrin (TPPS) in aqueous solutions have shown that the TPPS dications do not add counterions, whereas the influence of Cl- and Br- on fluorescence is realized according to the external heavy-atom effect mechanism. For nonaqueous o-TTP solutions with admixtures of HCl and CF3COOH the short-wavelength fluorescence from the S3 (B1) level has been detected whose unusual peculiarity is its enhancement in passing from 293 K to 77 K.


Population level policies on alcohol: are they still appropriate given that "alcohol is good for the heart'?
The research finding that a reduced risk of coronary heart disease is associated with alcohol use has been widely disseminated. This paper addresses the significance of this research finding for decisions about what kind of alcohol harm reduction policies it is appropriate to implement. The public policies of greatest concern to the vested interest groups are population-based policies such as taxation and restrictions on availability. Other public policies which impact on the drinkers' environment are minimum drinking age laws, drink drive deterrence, prevention of intoxication in public drinking places and restrictions on advertising. A review of the effectiveness of the available strategies provided strong support for environmentally focused approaches rather than education targeting high-risk drinkers. Many of these environmental strategies seem unlikely to have a significant direct effect of reducing "heart healthy' drinking by older people. Furthermore, the impact of alcohol on premature mortality among the older sectors of the population has to be weighed against any possible impact on younger drinkers resulting in loss of or disability in many years of life. Given the relative effectiveness of environmental strategies for reducing harm and the likely low level of impact of these strategies on the heart healthy drinking of older people it is concluded that the evidence of reduced risk of coronary heart disease does not preclude their implementation.


Adjuvant transarterial chemoembolization following radical resection for intrahepatic cholangiocarcinoma: A multi-center retrospective study.
Background and Aims: The prognosis of intrahepatic cholangiocarcinoma (ICC) after radical resection is far from satisfactory, but the effect of postoperative transarterial chemoembolization (p-TACE) remains controversial. This multi-center retrospective study was to evaluate the clinical value of p-TACE and identify the selected patients who would benefit from p-TACE. Methods: Data of ICC patients who underwent radical resection with/without p-TACE therapy was obtained from 12 hepatobiliary centers in China between Jan 2014 and Jan 2017. Overall survival (OS) was set as the primary endpoint, which was analyzed by the Kaplan-Meier method before and after propensity score matching (PSM). Subgroup analysis was conducted based on the established staging system and survival risk stratification. Results: A total of 335 patients were enrolled in this study, including 39 patients in the p-TACE group and 296 patients in the non-TACE group. Median OS in the p-TACE group was longer than that in the non-TACE group (63.0 months vs. 18.0 months, P=0.041), which was confirmed after 1:1 PSM (P=0.009). According to the 8th TNM staging system, patients with stage II and stage III stage would be benefited from p-TACE (P=0.021). Subgroup analysis stratified by risk factors showed that p-TACE could only benefit patients with risk factors <2 (P=0.027). Conclusion: Patients with ICC should be recommended to receive p-TACE following radical resection, especially for those with stage II, stage III or risk factors <2. However, the conclusion deserved further validation.


Comparison of Cold Technique Tonsillectomy and Thermal Welding Tonsillectomy at Different Age Groups.
The aim of this study is to compare objectively advantages and disadvantages of cold technique tonsillectomy and thermal welding tonsillectomy at the same case. A total of 100 patients, 53 patients younger than 12 years of age and 47 patients elder than 12 years of age, were included in this study. Tonsillectomy was performed by using cold technique on the right side of the palatine tonsils and thermal welding on the left side. Right and left sides were compared regarding perioperative bleeding, surgical dissection time, postoperative pain scale at the 1st and 7th day and postoperative bleeding parameters. Perioperative bleeding was found to be higher in cold technique side in patients younger than 12 years of age (p<0.001). Postoperative pain score on the day 1 was significantly higher in cold technique side, whereas it was found to be higher in thermal welding side at postoperative day 7 (p<0.001). Perioperative bleeding was found to be significantly higher in cold technique side (p<0.001) and surgical dissection time of thermal welding was found to be longer (p<0.001) in patients elder than 12 years of age. Postoperative pain score at the day 1 and day 7 was found to be higher in thermal welding side (p<0.001). Postoperative pain score at the day 1 and day 7 were statistically significantly higher in patients elder than 12 years of age. As a result, both techniques have its unique superior aspects and both can be applied as a routine tonsillectomy technique.


TAGADA: a scalable pipeline to improve genome annotations with RNA-seq data.
Genome annotation plays a crucial role in providing comprehensive catalog of genes and transcripts for a particular species. As research projects generate new transcriptome data worldwide, integrating this information into existing annotations becomes essential. However, most bioinformatics pipelines are limited in their ability to effectively and consistently update annotations using new RNA-seq data. Here we introduce TAGADA, an RNA-seq pipeline for Transcripts And Genes Assembly, Deconvolution, and Analysis. Given a genomic sequence, a reference annotation and RNA-seq reads, TAGADA enhances existing gene models by generating an improved annotation. It also computes expression values for both the reference and novel annotation, identifies long non-coding transcripts (lncRNAs), and provides a comprehensive quality control report. Developed using Nextflow DSL2, TAGADA offers user-friendly functionalities and ensures reproducibility across different computing platforms through its containerized environment. In this study, we demonstrate the efficacy of TAGADA using RNA-seq data from the GENE-SWiTCH project alongside chicken and pig genome annotations as references. Results indicate that TAGADA can substantially increase the number of annotated transcripts by approximately [Formula: see text] in these species. Furthermore, we illustrate how TAGADA can integrate Illumina NovaSeq short reads with PacBio Iso-Seq long reads, showcasing its versatility. TAGADA is available at github.com/FAANG/analysis-TAGADA.


Bedside electrical impedance tomography (EIT) for early assessment of Lung function in Liver transplantation.
Early extubation after liver transplantation is recommended by the Enhanced Recovery After Surgery Society (ERAS guideline 2022) as the benefit to avoid prolonged mechanical ventilation and associated complications. The timing of extubation is challenged by early postoperative hypoxia due to both preoperative and surgical status. Timely identifying the cause of hypoxia, typically including hepatopulmonary syndrome (HPS), pulmonary edema, atelectasis, and effusion is critical to prevent delayed extubation, especially in cases with HPS. As a radiation-free bedside approach, electrical impedance tomography (EIT) facilitates real-time lung ventilation and perfusion(V/Q) monitoring, however, the applicability of EIT V/Q monitoring for the assessment of HPS and its value for the management of hypoxia after liver transplantation has never been reported. Under this context, we took the lead in monitoring V/Q with EIT for a 12-year-old girl diagnosed with HPS who suffered intra and post liver transplantation hypoxia at the bedside. Our image raises key points for physicians that: 1) V/Q measurement of intrapulmonary shunts with bedside EIT holds the potential to identify HPS, a novel application that requires clinical validation. 2) EIT helps timely identify hypoxia after liver transplantation to support early individualized respiratory interventions. To our knowledge, V/Q monitoring by EIT is increasingly used in respiratory therapy, especially for ARDS PEEP titration, pulmonary embolism detection and lung transplantation assessment. However, The potential of EIT in liver transplant patients has been rarely studied. Our images might provide a novel and convenient approach to diagnosis HPS at bedside and offer new clinical applications for EIT imaging techniques.


Gravity induced fluid fragmentation in an asymmetric microfluidic Y junction.
In this study, the spontaneous and forced imbibition of an asymmetric Y junction by a completely wetting fluid submitted to the action of gravity is investigated. The considered junction is made of three capillaries of close but different dimensions. This system is used as a simple model to study gravity induced fluid fragmentation in microfluidic junctions and porous media. Using simple calculations, analytical results predict a fragmentation criterion, two dynamics leading to fluid fragmentation and asymptotically drop lengths and spacings. Both spontaneous and forced imbibitions of three microfluidic Y junctions were undertaken, varying the capillary and Bond numbers. Results show the validity of the fragmentation criterion, the derived junction imbibition dynamics and confirm qualitatively the analytical description of the fragmentation process. The dynamics prior to fragmentation does not depend on whether the imbibition is forced or spontaneous. However, it depends on the position with respect to gravity of the junction branch of lowest Laplace pressure. Finally, gravity induced fragmentation happens in the squeezing regime described in the Drop-On-Demand (DOD) literature at the junction and is locally controlled by viscosity rather than gravity.


High spatial resolution upgrade of the electron cyclotron emission radiometer for the DIII-D tokamak.
The 40-channel DIII-D electron cyclotron emission (ECE) radiometer provides measurements of Te(r,t) at the tokamak midplane from optically thick, second harmonic X-mode emission over a frequency range of 83-130 GHz. The frequency spacing of the radiometer's channels results in a spatial resolution of ∼1-3 cm, depending on local magnetic field and electron temperature. A new high resolution subsystem has been added to the DIII-D ECE radiometer to make sub-centimeter (0.6-0.8 cm) resolution Te measurements. The high resolution subsystem branches off from the regular channels' IF bands and consists of a microwave switch to toggle between IF bands, a switched filter bank for frequency selectivity, an adjustable local oscillator and mixer for further frequency down-conversion, and a set of eight microwave filters in the 2-4 GHz range. Higher spatial resolution is achieved through the use of a narrower (200 MHz) filter bandwidth and closer spacing between the filters' center frequencies (250 MHz). This configuration allows for full coverage of the 83-130 GHz frequency range in 2 GHz bands. Depending on the local magnetic field, this translates into a "zoomed-in" analysis of a ∼2-4 cm radial region. Expected uses of these channels include mapping the spatial dependence of Alfven eigenmodes, geodesic acoustic modes, and externally applied magnetic perturbations. Initial Te measurements, which demonstrate that the desired resolution is achieved, are presented.


Apoptosis-promoting properties of miR-3074-5p in MC3T3-E1 cells under iron overload conditions.
Iron overload can promote the development of osteoporosis by inducing apoptosis in osteoblasts. However, the mechanism by which miRNAs regulate apoptosis in osteoblasts under iron overload has not been elucidated. The miRNA expression profile in MC3T3-E1 cells under iron overload was detected by next generation sequencing. qRT-PCR was used to determine the expression of miR-3074-5p in MC3T3-E1 cells under iron overload. The proliferation of MC3T3-E1 cells was tested using CCK-8 assays, and apoptosis was measured using flow cytometry. The miRanda and TargetScan databases were used to predict the target genes of miR-3074-5p. Interaction between miR-3074-5p and the potential target gene was validated by qRT-PCR, luciferase reporter assay and western blotting. We found that iron overload decreased the cell viability and induced apoptosis of MC3T3-E1 cells. The results of next generation sequencing analysis showed that miR-3074-5p expression was significantly increased in MC3T3-E1 cells under iron overload conditions, which was confirmed by further experiments. The inhibition of miR-3074-5p attenuated the apoptosis of iron-overloaded MC3T3-E1 cells. Furthermore, the expression of Smad4 was decreased and was inversely correlated with miR-3074-5p expression, and overexpression of Smad4 partially reversed the viability inhibition of iron-overloaded MC3T3-E1 cells by relieving the suppression of ERK, AKT, and Stat3 phosphorylation, suggesting its regulatory role in the viability inhibition of iron-overloaded MC3T3-E1 cells. The luciferase reporter assay results showed that Smad4 was the target gene of miR-3074-5p. miR-3074-5p functions as an apoptosis promoter in iron-overloaded MC3T3-E1 cells by directly targeting Smad4.


Changes in prevalence of periodontitis in two German population-based studies.
We aimed to assess changes of periodontal status in Germany. The Studies of Health in Pomerania (SHIP) are two cross-sectional population-based studies conducted during 1997-2001 (SHIP-0, 20-81 years, n = 3736) and 2008-2012 (SHIP-Trend, 20-84 years, n = 3622) in northeast Germany. The German Oral Health Studies (DMS, 35-44 and 65-74 years) are national cross-sectional population-based surveys conducted in 1997 (DMS III, n = 1454) and 2005 (DMS IV, n = 1668), whose results were separately reported for West and East Germany. Prevalences, percentages and numbers of teeth affected were defined. In SHIP, prevalence of attachment loss (AL) ≥ 3 mm decreased from 89.7% (95% confidence interval (CI): 88.6-90.8) to 85.1% (95%CI: 83.9-86.3) (p < 0.05) and the mean extent reduced from 62.8% (95%CI: 61.7-63.8) to 55.9% (95%CI: 54.9-56.9) (p < 0.05). Probing depth (PD) ≥ 4 mm and the respective extent remained unchanged. In West Germany, AL ≥ 3 mm decreased for 35-44-year-olds and increased for 65-74-year-olds (p < 0.05). In SHIP and DMS, the number of teeth in dentates increased significantly in all age groups. Prevalences and extents of AL improved almost in all age categories in SHIP and West German adults, whereas PDs remained unchanged. Nonetheless, the improvement of periodontal conditions implies an increase of treatment needs regarding moderately diseased teeth because of simultaneous increases of the number of present teeth.


Comparison of post-operative lordosis with the PEEK cage and the cervical plate.
The maintenance of post-operative lordosis has been shown to be a key factor in decreasing adjacent level disc stress. Previous studies of the PEEK (polyether ketone) cage have used intervertebral bony fusion as the primary measure of surgical success; however, little is known about its effects on spinal curvature. Our objective was to compare the PEEK cage to the cervical plate with respect to the maintenance of cervical lordosis at one year. Secondary outcomes included fusion and complication rates. We performed a retrospective study of patients who underwent ACDF (anterior cervical discectomy and fusion) by two different methods; 13 patients were treated with the PEEK cage, and 22 with allograft and plating. Patient and treatment characteristics were similar in both groups. Average global lordotic curvature (C2-C7) was increased by 1.7 degrees for the PEEK cage and decreased by 1.6 degrees for the plate after an average follow-up of 12.46 and 14.95 months, respectively. Regional lordosis for the PEEK cage and plate was decreased by 2.5 and 2.1 degrees, respectively for the same time period. These differences did not achieve statistical significance. Bony fusion was observed in all patients. One patient in each group developed persistent mild dysphagia. The PEEK cage is comparable to the anterior cervical plate in the maintenance of post-operative cervical lordosis.


Direct inhibition of the cloned Kv1.5 channel by AG-1478, a tyrosine kinase inhibitor.
The action of tyrphostin AG-1478, a potent protein tyrosine kinase (PTK) inhibitor, on rat brain Kv1.5 channels (Kv1.5) stably expressed in Chinese hamster ovary cells was investigated using the whole cell patch-clamp technique. AG-1478 rapidly and reversibly inhibited Kv1.5 currents at 50 mV in a concentration-dependent manner with an IC50 of 9.82 microM. AG-1478 accelerated the decay rate of inactivation of Kv1.5 currents without modifying the kinetics of current activation. Pretreatment with the structurally dissimilar PTK inhibitors (genistein and lavendustin A) had no effect on the AG-1478-induced inhibition of Kv1.5 and did not modify the AG-1478-induced current kinetics. The rate constants for binding and unbinding of AG-1478 were 1.46 microM(-1) x s(-1) and 10.19 s(-1), respectively. The AG-1478-induced inhibition of Kv1.5 channels was voltage dependent, with a steep increase over the voltage range of channel opening. However, the inhibition exhibited voltage independence over the voltage range in which channels are fully activated. AG-1478 produced no significant effect on the steady-state activation or inactivation curves. AG-1478 slowed the deactivation time course, resulting in a tail crossover phenomenon. Inhibition of Kv1.5 by AG-1478 was use dependent. The present results suggest that AG-1478 acts directly on Kv1.5 currents as an open-channel blocker and independently of the effects of AG-1478 on PTK activity.


Closing the Mass Balance on Fluorine on Papers and Textiles.
Papers and textiles that are treated with per- and polyfluoroalkyl substances (PFASs) are sources of human and environmental exposure. Data for individual PFASs, such as perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA), are not placed into the context of total fluorine for papers and textiles. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to quantify volatile and ionic PFASs, respectively, and the total oxidizable precursor (TOP) assay was used to quantify precursors that form perfluoroalkyl carboxylates. Molar sums of PFASs obtained by GC-MS, LC-MS/MS, and precursors were compared to total fluorine (nmol F/cm2) determined by particle-induced gamma ray emission (PIGE) spectroscopy, measured before and after extraction. Volatile and ionic PFASs and unknown precursors accounted for 0-2.2%, 0-0.41%, and 0.021-14%, respectively, of the total nmol F/cm2 determined by PIGE. After extraction, papers and textiles retained 64 ± 28% to 110 ± 30% of the original nmol F/cm2 as determined by PIGE, indicating that the majority of fluorine remains associated with the papers and textiles. The sum of PFASs in the volatile, ionic, and precursor fraction, and total fluorine after extraction indicate that mass balance was achieved (within analytical error) of the initial total fluorine measured by PIGE.


A long-term frequency stabilized deep ultraviolet laser for Mg+ ions trapping experiments.
As many precision laser spectroscopy experiments require frequency stabilized lasers, development of long-term stabilized lasers is of great interest. In this work, we report long-term frequency stabilization of a 280 nm deep ultraviolet laser to a high precision wavemeter with digital servo control such that the long-term drift of the laser frequency was greatly reduced. Long-term laser frequency drift was measured with a fiber frequency comb system over 8 h. After locking, the maximum drift rate of the 280 nm laser was lowered from 576 MHz/h to 6.4 MHz/h. With proper environment control of the wavemeter, the maximum drift rate of the 280 nm laser was further lowered to less than 480 kHz/h. The locked laser system was successfully used in a Mg(+) ions trapping experiment, which was also discussed in this work.


Intergenerational associations of the approximate number system in toddlers and their parents.
From birth, humans are able to discriminate quantities using the approximate number system (ANS). However, previous methods have only been suitable to examine ANS functioning in infancy and older children. The goals of this study were twofold: first, to modify an existing method of assessing ANS functioning for toddlerhood; and second, to investigate individual differences in toddlers' ANS performance by examining correlations with their parents' ANS acuity. Using a preferential looking paradigm, we found that 1- to 3-year-olds (N = 46) looked significantly longer to numerically changing images compared to numerically constant ones suggesting that the paradigm is a suitable measure of ANS functioning in toddlerhood. Furthermore, we found a positive relation between toddlers' ANS performance and that of their parents (assessed using a non-symbolic number comparison task) independent of children's vocabulary or parents' perceived math ability or preference for math. These findings are consistent with a specific intergenerational transmission of the ANS. Statement of contribution What is already known on this subject? Past methods used to examine ANS functioning were only suitable for infants and older children. Little research has examined sources underlying individual difference in ANS acuity. What does this study add? We developed a preferential looking task to assess ANS functioning in toddlerhood. Individual differences in toddlers' ANS functioning are correlated with their parents' ANS acuity.


Functional Circuitry of the Retina.
The mammalian retina is an important model system for studying neural circuitry: Its role in sensation is clear, its cell types are relatively well defined, and its responses to natural stimuli-light patterns-can be studied in vitro. To solve the retina, we need to understand how the circuits presynaptic to its output neurons, ganglion cells, divide the visual scene into parallel representations to be assembled and interpreted by the brain. This requires identifying the component interneurons and understanding how their intrinsic properties and synapses generate circuit behaviors. Because the cellular composition and fundamental properties of the retina are shared across species, basic mechanisms studied in the genetically modifiable mouse retina apply to primate vision. We propose that the apparent complexity of retinal computation derives from a straightforward mechanism-a dynamic balance of synaptic excitation and inhibition regulated by use-dependent synaptic depression-applied differentially to the parallel pathways that feed ganglion cells.


Persistence of multidrug-resistant HIV-1 without antiretroviral treatment 2 years after sexual transmission.
To understand the virological mechanisms of 2-year persistence of multidrug-resistant virus without selective antiretroviral pressure in HIV-1-infected patients. Two patients were contaminated recently by their HIV-1-infected partners, who had received, before the transmission, all available antiretroviral drugs and who exhibited a severe therapeutic failure. The resistance mutations analysis was performed by clonal sequencing of 1.2 kb of pol gene in plasma of index and sources patients. Sequencing of HIV-1 DNA was performed in PBMCs of index patients. Genotypic testing performed in index patients at time of seroconversion showed resistance mutations to three classes of drugs. All mutations were linked on the same viral genome and all quasispecies carried all mutations. No wild-type virus was detected. The same results were found in source patients and showed that all mutations were transmitted. In the index patients, all mutations persisted over 2 years without antiretroviral treatment. Moreover, the resistance mutations were all archived in the cellular reservoir. Viral load and CD4 count of index patients remained unchanged during 2 years of follow-up. Only multidrug-resistant viruses were detected in the source patients and could be transmitted in index patients. In the latter, an expansion of predominant multidrug-resistant quasispecies and the 'archival' of all mutations were observed. These results explain the persistence of mutations and suggest that it is highly difficult to return to a wild-type viral population, sensitive to an antiretroviral treatment. The treatment of index patients is limited and the major risk is the transmission of these multidrug-resistant viruses. This work was presented in part in the XII International HIV Drug Resistance Workshop, Los Cabos, Mexico, June 2003; and in the 2nd IAS Conference on HIV Pathogenesis & Treatment, Paris, France, July 2003.


Spermidine enhances the silk production by mulberry silkworm.
Polyamines are ubiquitous low molecular weight polycationic aliphatic amines involved in diverse cellular processes. Spermidine (Spd), a polyamine, has been proved to be crucial for cell survival in various organisms. Our study reports the effect of Spd on the growth of Bombyx mori. Silkworms showed improved silk gland weight and economic parameters in the fifth instar larval stage when treated with different concentrations of Spd, in the range of 25-75 µM. The worms treated with Spd produced 31% more silk when compared with the control worms. Altogether, this study establishes that Spd-treated leaves can be fed into the larvae for better silk production.


Self-monitored blood pressure: a role in clinical practice?
Electronic self-monitoring of blood pressure is increasing in popularity and most international guidelines on the management of hypertension approve cautious use of the technique in the assessment of potentially hypertensive individuals. A recent editorial in the Archives of Internal Medicine suggested that it was "appropriate to encourage the widespread use of self recorded BP as an important adjunct to the clinical care of the patient with hypertension". Such a statement is based on increasing evidence that self-monitoring of blood pressure gives similar information to daytime ambulatory blood pressure -- a now well-established technology in the management of hypertension. Suggested strategies for the use of self-monitoring of blood pressure include monitoring in individuals whose clinical risk status is low enough that they need not necessarily be given medical therapy simply on the basis of a clinic pressure (i.e. at a 10 year risk of cardiovascular disease below 20%). The threshold for defining 'normotension/hypertension' is now regarded as being broadly similar for ABPM and SBPM and is set at 135/85 mmHg. In a recent meta-analysis of all available studies the average difference between these techniques, using the same patients, is -1.7/1.2 mmHg. There is some evidence that careful use of self-monitoring may improve blood pressure control in patients who are otherwise resistant to care. Self-monitoring of blood pressure has now been shown in at least one major prospective study to predict outcome better than clinic pressures and in that setting it now has equivalence to the use of ABPM. There remain issues regarding the availability of validated devices, the quality of training of patients in their use and the possibility that inaccurate recording might occur, either deliberately or by accident. Self-monitoring of blood pressure may well not give the same readings as carefully measured blood pressure by research nurses but its use is clearly superior to routine clinical practice. The technique is ripe for widespread application.


Identification and characterization of amino acid residues essential for the active site of UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from Staphylococcus aureus.
The enzymes essential for bacterial peptidoglycan biosynthesis are attractive targets for antimicrobial drug development. One of these is MurB, which contains FAD as a cofactor and catalyzes the NADPH-dependent reduction of UDP-N-acetylenolpyruvylglucosamine (UDP-GlcNAcEP) to UDP-N-acetylmuramic acid. This study examined the roles of the conserved amino acid residues of Staphylococcus aureus MurB, which are located near the active site in x-ray crystal structures. Seven of 11 site-directed mutated murB genes lost the ability to complement a temperature-sensitive S. aureus murB mutant. Biochemical characterization of the seven mutated MurB proteins revealed that they cannot carry out the reduction of UDP-GlcNAcEP, although they can all catalyze the intramolecular reduction of FAD via NADPH. Spectrometric analyses of the oxidized form of the mutated proteins in the presence and absence of NADP+ or UDP-GlcNAcEP revealed that these essential amino acid residues play four distinct roles in substrate interactions: Arg213 is essential for maintenance of the electronic state of FAD; Arg176 is required for interaction with UDP-GlcNAcEP; His259 is required for interaction with both UDP-GlcNAcEP and NADP+; and Asn71, Tyr175, Ser226, and Glu296 are not apparently required for interaction with either ligand. The results presented here identify for the first time the amino acid residues of MurB that are required for the interaction with UDP-Glc-NAcEP and NADP+.


The response of MRI contrast parameters in in vitro tissues and tissue mimicking phantoms to fractionation by histotripsy.
Histotripsy is a non-invasive, focused ultrasound lesioning technique that can ablate precise volumes of soft tissue using a novel mechanical fractionation mechanism. Previous research suggests that magnetic resonance imaging (MRI) may be a sensitive image-based feedback mechanism for histotripsy. However, there are insufficient data to form some unified understanding of the response of the MR contrast mechanisms in tissues to histotripsy. In this paper, we investigate the response of the MR contrast parameters R1, R2, and the apparent diffusion coefficient (ADC) to various treatment levels of histotripsy in in vitro porcine liver, kidney, muscle, and blood clot as well in formulations of bovine red blood cells suspended in agar gel. We also make a histological analysis of histotripsy lesions in porcine liver. We find that R2 and the ADC are both sensitive to ablation in all materials tested here, and the degree of response varies with tissue type. Correspondingly, under histologic analysis, the porcine liver exhibited various levels of mechanical disruption and necrotic debris that are characteristic of histotripsy. While the area of intact red blood cells and nuclei found within these lesions both decreased with increasing amounts of treatment, the area of red blood cells decreased much more rapidly than the area of intact nuclei. Additionally, the decrease in area of intact red blood cells saturated at the same treatment levels at which the response of the R2 saturated while the area of intact nuclei appeared to vary linearly with the response of the ADC.


Glycine receptor-mediated inhibition of medial prefrontal cortical pyramidal cells.
Using whole-cell patch clamp recording on medial prefrontal cortical slices of rats aged 17-33 postnatal days, we demonstrated the glycine-induced strychnine-sensitive outward currents. The amplitude of the peak current increased with the concentrations of glycine with an EC50 of 74.7 μM. Application of 1μM strychnine alone to cells caused a slight inward current without blocking the sIPSCs, indicating that GlyRs in the mPFC are activated by an endogenous ligand that can be released tonically. Glycine reversibly depressed firing rate in cells from both layer 6 and layer 3, with significantly greater inhibition on the former than the latter (EC50 12.9 vs 85.6 μM). Glycine hyperpolarized membrane potential in cells of both layer 6 and layer 3 depending on its concentrations, with an IC50 of 99.1 and 207.2 μM, respectively. We propose that GlyRs participate in a novel inhibitory mechanism in mPFC, modulating neuronal activity. This finding further supports an important role of GlyR in cortical function and dysfunction.


Characterization of Recombinant Adeno-Associated Viral Transduction and Safety Profiles in Cardiomyocytes.
Cardiovascular diseases (CVD) are the leading causes for human mortality. However, the effective treatment for these diseases are still lacking. Currently, gene therapy could be a potential way for efficiently treating heart diseases. The aim of our study is to analyze the transduction efficacy and safety profile of recombinant adeno associated virus (AAV) serotype 9 for cardiomyocytes in vivo and in vitro. We produced rAAV serotype 9 expressing enhanced green fluorescence protein (EGFP) driven by a cardiac troponin T (cTNT) promoter, and characterized its transduction efficiency in primary cultured cardiomyocytes in vitro, and in wild-type mouse heart tissue in vivo. Our data showed that rAAV9 efficiently transduced mouse cardiomyocytes in vitro. Following intravenous injection, rAAV9 could efficiently and safely transduce cardiomyocytes that are involved in heart diseases. Our findings suggested that rAAV9 can efficiently and safely transduce cardiomyocytes in vitro and/or in vivo. The rAAV9 serotype vector could constitute a powerful toolbox for future gene therapy of heart diseases.


Influence of tetracyclines on human polymorphonuclear leukocyte function.
Low concentrations of oxytetracycline, doxycycline, or minocycline (less than 10 micrograms/ml) did not influence in vitro polymorphonuclear leukocyte random migration, chemiluminescence, or glucose oxidation. At high concentrations of doxycycline or minocycline (greater than 10 micrograms/ml), chemiluminescence and glucose oxidation were impaired. High concentrations of doxycycline also reduced random migration. Oxytetracycline did not influence these functions in concentrations up to 100 micrograms/ml. The inhibiting effect of doxycycline and minocycline was abolished when 4 mM Mg2+ was added to the reaction mixture, and 4 mM Ca2+ partly restored minocycline-inhibited polymorphonuclear leukocyte functions. This indicates that the major effect of tetracyclines on in vitro polymorphonuclear leukocyte functions is mediated by their divalent cation chelating effect and that the results of in vitro experiments are highly dependent on the concentration of divalent cations in the reaction mixtures. The difference between the tetracyclines may be due to differences in lipid solubility, with solubility being highest for minocycline and lowest for oxytetracycline, or to different divalent cation chelating ability.


Investigation on Inter-Limb Coordination and Motion Stability, Intensity and Complexity of Trunk and Limbs during Hands-Knees Crawling in Human Adults.
This study aimed to investigate the inter-limb coordination pattern and the stability, intensity, and complexity of the trunk and limbs motions in human crawling under different speeds. Thirty healthy human adults finished hands-knees crawling trials on a treadmill at six different speeds (from 1 km/h to 2.5 km/h). A home-made multi-channel acquisition system consisting of five 3-axis accelerometers (ACC) and four force sensors was used for the data collection. Ipsilateral phase lag was used to represent inter-limb coordination pattern during crawling and power, harmonic ratio, and sample entropy of acceleration signals were adopted to depict the motion intensity, stability, and complexity of trunk and limbs respectively. Our results revealed some relationships between inter-limb coordination patterns and the stability and complexity of trunk movement. Trot-like crawling pattern was found to be the most stable and regular one at low speed in the view of trunk movement, and no-limb-pairing pattern showed the lowest stability and the greatest complexity at high speed. These relationships could be used to explain why subjects tended to avoid no-limb-pairing pattern when speed was over 2 km/h no matter which coordination type they used at low speeds. This also provided the evidence that the central nervous system (CNS) chose a stable inter-limb coordination pattern to keep the body safe and avoid tumbling. Although considerable progress has been made in the study of four-limb locomotion, much less is known about the reasons for the variety of inter-limb coordination. The research results of the exploration on the inter-limb coordination pattern choice during crawling from the standpoint of the motion stability, intensity, and complexity of trunk and limbs sheds light on the underlying motor control strategy of the human CNS and has important significance in the fields of clinical diagnosis, rehabilitation engineering, and kinematics research.


Strategies to Avoid Underdiagnosing Pit-and-Fissure Caries.
Traditionally, testing for whether pit-and-fissure caries should be restored involved probing with a sharp explorer and evaluating resistance to withdrawal (ie, "stick"). Alternative visual methods of evaluation and classification have been proposed, validated, and accepted formally in the core curriculum on caries management in both Europe and North America. This article examines the resistance to occlusal breakdown of fluoridehardened enamel despite progression of underlying dentin caries with accompanying difficulty in diagnosis. Traditional methods of pit-and-fissure caries diagnosis, including radiographs or fissure probing with an explorer, have been shown to be inaccurate and potentially destructive. The clinical process of using the visual/tactile International Caries Detection and Assessment System (ICDAS) and/or the Caries Classification System (CCS) is described and illustrated through case examples.


pH dependence of melanoma cell migration: protons extruded by NHE1 dominate protons of the bulk solution.
Migration and morphology of human melanoma cells (MV3) depend on extracellular pH (pHe) and the activity of the Na+/H+ exchanger NHE1. To distinguish effects of NHE1 activity per se from effects of pHe we compared an NHE1-deficient mutant with rescued and wild-type cells. Time lapse video microscopy was used to investigate migratory and morphological effects caused by pHe and NHE1 activity, and a membrane-bound fluorescein conjugate was employed for ratiometric pH measurements at the outer leaflet of the cell membrane. As long as NHE1 remained inactive due to deficiency or inhibition by cariporide (HOE642) neither migration nor morphology was affected by changes in pHe. Under these conditions pH at the outer leaflet of the plasma membrane was uniform all over the cell surface. The typical pH dependence of MV3 cell migration and morphology could be reconstituted by restoring NHE1 activity. At the same time the proton gradient at the outer leaflet of the plasma membrane with the higher proton concentration at the leading edge and the lower one at the cell rear was re-established as well. Hence, NHE1 activity generates a proton gradient at the cell surface accompanied by the cells' ability to respond to changes in pHe (bulk pH). We conclude that NHE1 activity contributes to the generation of a well-defined cell surface pH by creating a proton gradient at the outer leaflet of the plasma membrane that is needed for (i) the development of a variety of morphologies including a distinct polarity and (ii) migration. A missing proton gradient at the cell surface cannot be compensated for by varying pHe.


Selective inhibition of nonspecific eosinophil staining or identification of eosinophilic granulocytes by paired counterstaining in immunofluorescence studies.
Nonspecific staining of eosinophilic granulocytes produced by nonimmune rabbit IgG conjugates, highly labeled with fluorescein isothiocyanate (FITC) or tetramethylrhodamine isothiocyanate (MRITC), was efficiently inhibited by pretreatment of tissue sections with diaminobenzidine and hydrogen peroxide; the brown reaction product developed in the presence of this substrate by the endogenous peroxidase of the granulocytes sheltered these cells from subsequent nonspecific staining without interfering with specific immunofluorescence staining of other tissue elements. Similar results were obtained with the alternative noncarcinogenic peroxidase substrate, p-phenylenediamine-pyrocatechol. Alternatively, the eosinophils could be identified by their nonspecific staining with FITC-labeled bovine serum albumin when this conjugate was combined with an MRITC conjugate used for specific staining; optical conditions selective for green and red fluorescence discriminated distinctly between double-stained eosinophils and purely red specifically stained cells.


The standard data model approach to patient record transfer.
This paper develops an approach to electronic data exchange of patient records from Ambulatory Encounter Systems (AESs). This approach assumes that the AES is based upon a standard data model. The data modeling standard used here is IDEFIX for Entity/Relationship (E/R) modeling. Each site that uses a relational database implementation of this standard data model (or a subset of it) can exchange very detailed patient data with other such sites using industry standard tools and without excessive programming efforts. This design is detailed below for a demonstration project between the research-oriented geriatric clinic at the Baltimore Veterans Affairs Medical Center (BVAMC) and the Laboratory for Healthcare Informatics (LHI) at the University of Maryland.


Renal neuronal nitric oxide synthase protein expression as a marker of renal injury.
Animal studies suggest that nitric oxide deficiency occurs in the remnant after 5/6 removal of renal mass. The present studies investigated the time course in relation to progression of renal disease, as well as the impact on individual renal nitric oxide synthase (NOS) isoforms. Rats were studied from 2 to 11 weeks after 5/6 ablation/infarction (A/I) of renal mass, with acceleration of progression by high protein and salt intake, in some groups. Measurements were made before sacrifice of 24-hour protein and creatinine excretion, blood was taken for creatinine and blood urea nitrogen (BUN) determination and the kidneys were investigated histologically for structural damage, abundance of endothelial NOS (eNOS) and neuronal NOS (nNOS), and in some groups for in vitro NOS activity. A time-dependent fall in glomerular filtration rate (GFR) and rise in proteinuria and glomerular sclerosis developed after 5/6 A/I. The nNOS abundance in cortex and medulla was decreased relative to shams, in all but the mildest injury and there was a strong, steep correlation between level of glomerular sclerosis and the degree of reduction in renal nNOS. Where measured, cortical NOS activity correlated with the nNOS abundance. In contrast, the eNOS abundance was either increased or unchanged in rats post A/I. Renal nNOS abundance was reduced in the 5/6 A/I model of renal disease when plasma creatinine> approximately 1 mg/dL and when> approximately 20% of remaining glomeruli were sclerosed.


Impact of female obesity and assisted reproduction on uncomplicated pregnancies and healthy births: a study of 428 336 births in Flanders.
What is the impact of BMI on uncomplicated pregnancies and healthy births in women who did or did not have medically assisted reproduction (MAR, i.e. ART or hormonal stimulation without manipulation of eggs or embryos) in the Flanders region (Belgium)? Women with a higher BMI who use MAR are at the highest risk of pregnancy and birth complications. Medically assisted reproduction (MAR) is used increasingly worldwide and is associated with increased risk of adverse perinatal outcomes. Obesity is also increasing globally and obese women are more likely to seek MAR since obesity is associated with infertility. When obese women undergo MAR, the risk of adverse outcomes may be enhanced but it is not clear to what extent. We conducted a registry-based study using the data from the Study Centre for Perinatal epidemiology database for years 2009-2015, region of Flanders, Belgium. This included 428 336 women. The average age was 30.0 years (SD 4.78), 194 061 (45.31%) were nulliparous, and 6.3% (n = 26 971) conceived with MAR. We examined the association of BMI and MAR with the following composite primary outcomes: 'uncomplicated pregnancy and birth' and 'healthy baby'. We conducted Poisson regression and adjusted for maternal age, parity, gestational weight gain, smoking and previous caesarean section. In our study, 36.80% (n = 157 623) of women had an uncomplicated pregnancy and birth according to the definition used. The predicted probability of having an uncomplicated pregnancy and birth for women with a BMI of 25 kg/m2 who conceived spontaneously was 0.33 (0.32 to 0.35), while it was 0.28 (0.24 to 0.32) for women who used hormonal stimulation and 0.26 (0.22 to 0.29) for women who used IVF/ICSI. This probability reduced with increasing BMI category for both MAR and non-MAR users. For women with a BMI of 30 kg/m2, the predicted probability of having an uncomplicated pregnancy and birth was 0.28 (0.26 to 0.30) for women who conceived spontaneously, and 0.22 (0.16 to 0.29) and 0.20 (0.14 to 0.26) for women who used hormonal stimulation only or IVF/ICSI, respectively. The predicted probability of having a healthy baby for women with a BMI of 25 kg/m2 who conceived spontaneously was 0.92 (0.91 to 0.93), 0.89 (0.87 to 0.92) for women who used hormonal stimulation only and 0.85 (0.84 to 0.87) for women who used IVF/ICSI. The database did not include data on socio-economic status, pre-pregnancy morbidities and paternal BMI. Subsequently, we could not adjust for these factors in the analysis. Obese women who use MAR are at the highest risk of pregnancy and birth complications. This increase in interventions also has cost and resource implications which is relevant for funding policies. Weight loss interventions prior to MAR seem plausible but their (cost-) effectiveness needs urgent investigation. F.W. received an Erasmus Plus training grant to visit A.B., L.A. and R.D. and conducted this study during this visit. The authors have no competing interest to declare. N/A.


Evaluation of New Monoclonal Anti-MyoD1 (MX049) for the Diagnosis of Rhabdomyosarcoma: Comparison with 5.8A, EP212, Anti-Desmin, Anti-Myogenin, and Fluorescence in situ Hybridization.
Rhabdomyosarcoma (RMS) is a primitive embryonal mesenchymal neoplasm demonstrating skeletal muscle differentiation. Diagnosis of RMS remains difficult due to the diversity of clinical features, pathological forms, and lesion's locations. Immunohistochemistry and Fluorescence in Situ Hybridization are common methods used to aid RMS diagnosis. In this research we tested protein expression of Desmin (Clone MX046), MyoD1 (Clone MX049), MyoD1 (Clone 5.8A), MyoD1 (Clone EP212), Myogenin (Clone F5D), and cytogenetic features in 21 RMS cases, with following results: positive rates of Desmin (Clone MX046), MyoD1 (Clone MX049), MyoD1 (Clone 5.8A), MyoD1 (Clone EP212) and Myogenin (Clone F5D) were 100.00%, 100.00%, 90.48%, 95.24% and 85.71%, respectively, with cytoplasmic stains of MyoD1 (Clone 5.8A) in 38.10% (8/21) cases and only nuclear stains of MyoD1 (Clone EP212), MyoD1 (Clone MX049) in all positive cases. FOXO1 gene was detected apart in 9 alveolar RMS samples, where MyoD1 (Clone MX049), MyoD1 (Clone 5.8A) and MyoD1 (Clone EP212) were 100% positive but MyoD1 (Clone 5.8A) only 44.44% (4/9). Thus we believe MyoD1 (Clone MX049) performs more sensitive and specific than MyoD1 (Clone 5.8A) and MyoD1 (Clone EP212).


Prehospital noninvasive pressure support ventilation for acute cardiogenic pulmonary edema.
Severe acute cardiogenic pulmonary edema (ACPE) can successfully be treated with noninvasive pressure support ventilation (NIPSV) in a clinical setting. Whether prehospital NIPSV starting early at patients' home and being continued until hospital arrival is feasible and improves ACPE emergency care is examined in this study. End points of the study were oxygen saturation at hospital admission and clinical outcome. Twenty-three patients suffering from severe cardiac pulmonary edema with severe dyspnea, an oxygen saturation of less than 90% and basal rales were included in this controlled prospective randomized trial. All patients received standard medical treatment and 10 patients were additionally treated with NIPSV (pressure support level, 12 cmH2O; positive endexpiratory pressure, 5 cmH2O; FiO2, 0.6) whereas the other patients received oxygen (8 l/min) via Venturi face mask. Improvement in oxygen saturation was significantly faster in the NIPSV group and oxygen saturation was higher at the time of the hospital admission (NIPSV=97.3+/-0.8%; standard=89.5+/-2.7%, P=0.002). A trend toward higher troponin T levels was seen in the standard treatment group. The need for intensive care treatment did not differ, and one patient of each treatment group died in hospital. No complications were noted during the treatment with NIPSV. Prehospital NIPSV is feasible and able to improve emergency management of ACPE.


Intervening sequences in 16S rRNA genes of Campylobacter sp.: diversity of nucleotide sequences and uniformity of location.
We found and sequenced intervening sequences (IVSs) in the PCR-amplicons of 16S rRNA genes of 3 strains of Campylobacter rectus, 2 strains of C. curvus and 2 strains of C. sputorum. The lengths of the IVSs were 140 to 233 bp. The IVSs of C. rectus were identical and had a sequence homology of 55 to 79% against those of C. curvus and C. helveticus. The IVSs of C. sputorum were 97.9-100% homologous but poorly homologous to the other IVSs. In spite of the diversities of the lengths and the nucleotide sequences, all of the IVSs were located at the same position in the 16S rRNA genes.


On the implementation of dose-volume objectives in gradient algorithms for inverse treatment planning.
A method that allows a straightforward implementation of dose-volume constraints in gradient algorithms for inverse treatment planning is presented. The method is consistent with the penalty function approach, which requires the formulation of an objective function with penalty terms proportional to the magnitudes of constraint violations. Dose constraints with respect to minimum and maximum target dose levels are incorporated in quadratic, dose-penalty terms. Analogously, quadratic volume-penalty terms in the objective function reflect the violation of dose-volume constraints imposing limits on the fractions of healthy organ volumes that can be irradiated above specified dose levels. It has been demonstrated that within the framework of this formulation neither modified objective functions nor finite difference gradient calculations are necessary for the incorporation of gradient minimization algorithms. As an example, a simple steepest descent algorithm is presented along with its application to illustrate prostate and lung cases.


Carcinocythemia (carcinoma cell leukemia). Report of two cases with English literature review.
Cancer cells of nonhematologic origin are rarely seen on routinely prepared peripheral blood smears. This report describes two patients with disseminated carcinoma, in near terminal state, one with malignant melanoma and another with transitional cell carcinoma of the ureteropelvic junction who developed carcinocythemia (CCA). A review of the English literature revealed only eight other cases in which CCA was documented. Four patients had a small cell bronchogenic ("oat cell") carcinoma, two had disseminated breast carcinoma and one each had advanced adenocarcinoma of ileum and abdominal rhabdomyosarcoma. The circulating malignant cells usually accounted for less than 10% of the nucleated blood cells, but in one patient comprised up to 80% of the leukocyte count. The mean time between detection of CCA and death was 5 weeks (median, 2 weeks) reflecting the fact that CCA is generally a terminal event.


It's What and When You Eat: An Overview of Transcriptional and Epigenetic Responses to Dietary Perturbations in Pancreatic Islets.
Our ever-changing modern environment is a significant contributor to the increased prevalence of many chronic diseases, and particularly, type 2 diabetes mellitus (T2DM). Although the modern era has ushered in numerous changes to our daily living conditions, changes in "what" and "when" we eat appear to disproportionately fuel the rise of T2DM. The pancreatic islet is a key biological controller of an organism's glucose homeostasis and thus plays an outsized role to coordinate the response to environmental factors to preserve euglycemia through a delicate balance of endocrine outputs. Both successful and failed adaptation to dynamic environmental stimuli has been postulated to occur due to changes in the transcriptional and epigenetic regulation of pathways associated with islet secretory function and survival. Therefore, in this review we examined and evaluated the current evidence elucidating the key epigenetic mechanisms and transcriptional programs underlying the islet's coordinated response to the interaction between the timing and the composition of dietary nutrients common to modern lifestyles. With the explosion of next generation sequencing, along with the development of novel informatic and -omic approaches, future work will continue to unravel the environmental-epigenetic relationship in islet biology with the goal of identifying transcriptional and epigenetic targets associated with islet perturbations in T2DM.


Structure and reactivity of homoleptic samarium(II) and thulium(II) phospholyl complexes.
Potassium 2,5-di-tert-butyl-3,4-dimethylphospholide K(dtp) (9) was synthesised in 45 % yield from commercially available starting materials by using zirconacyclopentadiene chemistry. Reaction of the K salt of this bulky anion and of the previously described potassium 2,5-bis(trimethylsilyl)-3,4-dimethylphospholide K(dsp) (8) with SmI(2) in diethyl ether afforded the homoleptic samarium(II) complexes 7 and 6, respectively, whose solid-state structures, [[Sm(dtp)(2)](2)] (7 a) and [[Sm(dsp)(2)](2)] (6 a), are dimeric owing to coordination of the phosphorus lone pairs to samarium, as shown by X-ray crystallography. Reaction of 8 with TmI(2) in diethyl ether afforded [Tm(dsp)(2)(Et(2)O)], which could not be desolvated without decomposition. In contrast, the coordinated ether group of the solvate [Tm(dtp)(2)(Et(2)O)], obtained from 9 and TmI(2), could easily be removed by evaporation of the solvent and extraction with pentane at room temperature, and the monomer [Tm(dtp)(2)] (5) could be isolated and was characterised by X-ray crystallography. Presumably, steric crowding in 5 is too high for dimerisation to occur. Compound 5, the first Tm(II) homoleptic sandwich complex, is remarkably stable at room temperature in solution and did not noticeably react with nitrogen, in sharp contrast with other thulium(II) species. As expected, 5, 6 and 7 all reacted with azobenzene to give the trivalent complexes [Tm(dtp)(2)(N(2)Ph(2))] (13), [Sm(dsp)(2)(N(2)Ph(2))], (14) and [Sm(dtp)(2)(N(2)Ph(2))] (15), respectively; 13 and 14 were characterised by X-ray crystallography. Complex 5 immediately reacted with triphenylphosphane sulfide at room temperature to give [[Tm(dtp)(2)](2)(mu-S)] (16), which was characterised by X-ray crystallography, whereas samarium(II) complexes 6 and 7 did not noticeably react with Ph(3)PS over 24 h under the same conditions.


(-)-Epicatechin-3-O-β-D-allopyranoside from Davallia formosana, Prevents Diabetes and Hyperlipidemia by Regulation of Glucose Transporter 4 and AMP-Activated Protein Kinase Phosphorylation in High-Fat-Fed Mice.
The purpose of this experiment was to determine the antidiabetic and lipid-lowering effects of (-)-epicatechin-3-O-β-D-allopyranoside (BB) from the roots and stems of Davallia formosana in mice. Animal treatment was induced by high-fat diet (HFD) or low-fat diet (control diet, CD). After eight weeks of HFD or CD exposure, the HFD mice were treating with BB or rosiglitazone (Rosi) or fenofibrate (Feno) or water through gavage for another four weeks. However, at 12 weeks, the HFD-fed group had enhanced blood levels of glucose, triglyceride (TG), and insulin. BB treatment significantly decreased blood glucose, TG, and insulin levels. Moreover, visceral fat weights were enhanced in HFD-fed mice, accompanied by increased blood leptin concentrations and decreased adiponectin levels, which were reversed by treatment with BB. Muscular membrane protein levels of glucose transporter 4 (GLUT4) were reduced in HFD-fed mice and significantly enhanced upon administration of BB, Rosi, and Feno. Moreover, BB treatment markedly increased hepatic and skeletal muscular expression levels of phosphorylation of AMP-activated (adenosine monophosphate) protein kinase (phospho-AMPK). BB also decreased hepatic mRNA levels of phosphenolpyruvate carboxykinase (PEPCK), which are associated with a decrease in hepatic glucose production. BB-exerted hypotriglyceridemic activity may be partly associated with increased mRNA levels of peroxisome proliferator activated receptor α (PPARα), and with reduced hepatic glycerol-3-phosphate acyltransferase (GPAT) mRNA levels in the liver, which decreased triacylglycerol synthesis. Nevertheless, we demonstrated BB was a useful approach for the management of type 2 diabetes and dyslipidemia in this animal model.


Enhanced expressions and histological characteristics of intravenously administered plasmid DNA in rat lung.
Cationic liposome-mediated gene transfection is a promising method for gene therapy. In this study, the transfection efficiency and histological patterns were evaluated in rat lung after intravenous administration via femoral vein of naked plasmid DNA, naked plasmid DNA with pretreatment of DOTAP, and DOTAP-cholesterol-plasmid DNA complex. Plasmid DNA encoding bacterial LacZ gene was used. For quantification of LacZ gene expression, beta-galactosidase assay was performed. For histologic examination, X-gal staining and immunohistochemical staining for transfected gene products were performed. Pretreatment of DOTAP prior to the infusion of naked plasmid DNA increased transfection efficiency up to a level comparable to DOTAP-cholesterol-plasmid DNA complex injection. Transfected genes were mainly expressed in type II pneumocytes and alveolar macrophages in all animals. We conclude that the high transfection efficiency is achievable by intravenous administration of naked plasmid DNA with pretreatment of DOTAP, to a level comparable to DOTAP-cholesterol-plasmid DNA complex. In this regard, naked plasmid DNA administration with pretreatment of DOTAP could be a more feasible option for intravenous gene transfer than DOTAP-cholesterol-plasmid DNA complex, in that the former is technically easier and more cost-effective than the latter with a comparable efficacy, in terms of intravenous gene delivery to the lung.


Elucidating Therapeutic and Biological Potential of Berberis baluchistanica Ahrendt Bark, Leaf, and Root Extracts.
Berberis baluchistanica Ahrendt is a medicinal plant known to have potential for the treatment of various diseases. In the present study, the ethanolic extracts of the bark, leaves, and roots of B. baluchistanica plant were evaluated for in vitro antimicrobial, anti-leishmanial, anticancer, and anti-inflammatory activities. The antibacterial and antifungal activities were determined by agar mix and agar well diffusion method. All extracts showed potential activity against the target bacteria (Bacillus subtilis, Bacillus licheniformis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Rhodococcus erythropolis, Salmonella typhi, and Staphylococcus aureus) and fungal strains (Aspergillus flavus, Aspergillus niger, and Mucor mucedo). S. aureus proved to be the most sensitive strain for each extract, with a maximum zone of inhibition for bark at 23 ± 0.12 mm, for leaves at 22 ± 0.36 mm, and for root extracts at 20.21 ± 0.06 mm). The minimum inhibitory concentration values of B. baluchistanica bark, leaves, and roots for different target bacterial strains ranged from 1.56 to 25 mg ml-1, and the minimum bactericidal concentrations were in the range of 3.12 to 25 mg ml-1, respectively. The root extract possessed potent antifungal activity against A. flavus with 83% of growth inhibition, A. niger with 80%, and M. mucedo with 73%. The bark extract was found active against M. mucedo with 86% of inhibition, followed by 70% against A. flavus and 60% against A. niger. The leaf extract showed a significant response by 83% inhibition against M. mucedo, followed by A. flavus and A. niger with 73 and 72% inhibition, respectively. In an anti-leishmanial bioassay, the inhibitory concentration (IC50) was observed for each extract against Leishmania major. The bark showed good activity (IC50 = 4.95 ± 0.36 mg/ml), followed by the roots (IC50 = 7.07 ± 0.18 mg/ml) and the leaves (IC50 = 8.25 ± 0.29 mg/ml). An evaluation of anticancer activity was done by using MTT cell assay against HeLa cell line. Upon comparing the values of each extract to the standard, it was revealed that the ethanolic bark extract showed the highest anticancer activity with IC50 = (12 ± 0.15 μg/ml), followed by the roots (14 ± 0.15 μg/ml) and the leaves (17 ± 0.21 μg/ml), respectively. The anti-inflammatory assay was undertaken by the inhibition of albumin denaturation activity, proteinase inhibitory activity, and heat-induced hemolysis activity. The IC50 value for protein denaturation of the bark was IC50 = 0.64 ± 0.25 mg/ml, followed by the roots (0.67 ± 0.21 mg/ml) and the leaves (0.73 ± 0.13 mg/ml). The proteinase inhibitory activity of the bark extract was IC50 = 0.55 ± 0.12 mg/ml, followed by the leaves (0.62 ± 0.23 mg/ml) and the roots (0.69 ± 0.15 mg/ml), respectively. For heat-induced hemolysis assay, the bark showed the lowest IC50 value (0.48 ± 0.15 mg/ml) compared to the leaves (0.52 ± 0.35 mg/ml) and the roots (0.58 ± 0.05 mg/ml) of the plant. All analyzed parts of the B. baluchistanica plant showed significant biological activities which make the plant medicinally important and a good candidate for the isolation of antimicrobial, inflammatory, and anticancer compounds. Further studies may lead us to determine the active compounds responsible for the biological activities of the plant extracts.


High pressure reactivity of propene by first principles molecular dynamics calculations.
The reactivity of propene under high pressure has been investigated in the framework of Car-Parrinello molecular dynamics. Changes in structural and electronic properties due to pressure have been analyzed in systems with a density ranging from 0.855 to 2.151 g/cm(3). A ionic collective mechanism which leads to the formation of oligomers has been found by both spin restricted and spin polarized formalism. The maximally localized Wannier centers analysis has allowed us to characterize the addition scheme and to identify a Wannier center with a high spread value involved in the formation of the principal reaction products.


Dextroplantation of a reduced left lateral section graft in an infant undergoing living donor liver transplantation.
Graft size matching is essential for successful liver transplantation in infant recipients. We present our technique of graft dextroplantation used in an infant who underwent living donor liver transplantation (LDLT) using a reduced left lateral section (LLS) graft. The patient was an 11-month-old female infant weighing 7.8 kg with hepatoblastoma. She was partially responsive to systemic chemotherapy. Thus, LDLT was performed to treat the tumor. The living donor was a 34-year-old mother of the patient. After non-anatomical size reduction, the weight of the reduced LLS graft was 235 g, with a graft-to-recipient weight ratio of 3.0%. Recipient hepatectomy was performed according to the standard procedures of pediatric LDLT. At the beginning of graft implantation, the graft was temporarily placed at the abdomen to determine the implantation location. The graft portal vein was anastomosed with an interposed external iliac vein homograft. As the liver graft was not too large and it was partially accommodated in the right subphrenic fossa, thus the abdominal wall wound was primarily closed. The patient recovered uneventfully. An imaging study revealed deep accommodation of the graft within the right subphrenic fossa. The patient has been doing well for six months without any vascular complications. This case suggests that dextroplantation of a reduced LLS graft can be a useful technical option for LDLT in infant patients.


Aluminum neurotoxicity in preterm infants receiving intravenous-feeding solutions.
Aluminum, a contaminant of commercial intravenous-feeding solutions, is potentially neurotoxic. We investigated the effect of perinatal exposure to intravenous aluminum on the neurologic development of infants born prematurely. We randomly assigned 227 premature infants with gestational ages of less than 34 weeks and birth weights of less than 1850 g who required intravenous feeding before they could begin enteral feeding to receive either standard or specially constituted, aluminum-depleted intravenous-feeding solutions. The neurologic development of the 182 surviving infants who could be tested was assessed by using the Bayley Scales of Infant Development at 18 months of age. The 90 infants who received the standard feeding solutions had a mean (+/-SD) Bayley Mental Development Index of 95+/-22, as compared with 98+/-20 for the 92 infants who received the aluminum-depleted solutions (P=0.39). In a planned subgroup analysis of infants in whom the duration of intravenous feeding exceeded the median and who did not have neuromotor impairment, the mean values for the Bayley Mental Development Index for the 39 infants who received the standard solutions and the 41 infants who received the aluminum-depleted solutions were 92+/-20 and 102+/-17, respectively (P=0.02). The former were significantly more likely (39 percent, vs. 17 percent of the latter group; P=0.03) to have a Mental Development Index of less than 85, increasing their risk of subsequent educational problems. For all 157 infants without neuromotor impairment, increasing aluminum exposure was associated with a reduction in the Mental Development Index (P=0.03), with an adjusted loss of one point per day of intravenous feeding for infants receiving the standard solutions. In preterm infants, prolonged intravenous feeding with solutions containing aluminum is associated with impaired neurologic development.


Comparison of multilevel modeling and the family-based association test for identifying genetic variants associated with systolic and diastolic blood pressure using Genetic Analysis Workshop 18 simulated data.
Identifying genetic variants associated with complex diseases is an important task in genetic research. Although association studies based on unrelated individuals (ie, case-control genome-wide association studies) have successfully identified common single-nucleotide polymorphisms for many complex diseases, these studies are not so likely to identify rare genetic variants. In contrast, family-based association studies are particularly useful for identifying rare-variant associations. Recently, there has been some interest in employing multilevel models in family-based genetic association studies. However, the performance of such models in these studies, especially for longitudinal family-based sequence data, has not been fully investigated. Therefore, in this study, we investigated the performance of the multilevel model in the family-based genetic association analysis and compared it with the conventional family-based association test, by examining the powers and type I error rates of the 2 approaches using 3 data sets from the Genetic Analysis Workshop 18 simulated data: genome-wide association single-nucleotide polymorphism data, sequence data, and rare-variants-only data. Compared with the univariate family-based association test, the multilevel model had slightly higher power to identify most of the causal genetic variants using the genome-wide association single-nucleotide polymorphism data and sequence data. However, both approaches had low power to identify most of the causal single-nucleotide polymorphisms, especially those among the relatively rare genetic variants. Therefore, we suggest a unified method that combines both approaches and incorporates collapsing strategy, which may be more powerful than either approach alone for studying genetic associations using family-based data.


Effect of interval between inseminations when using frozen-thawed boar sperm on fertility and fetal paternity in mature gilts.
Multiple artificial inseminations (AI) are used in swine to compensate for variation in time of ovulation and following each insemination, a uterine immune response occurs to remove dead and damaged sperm. There are more dead and damaged sperm inseminated with use of frozen-thawed-semen (FTS) and closer insemination intervals are required to compensate for the reduced lifespan of the sperm. Therefore, the objectives of this experiment were to evaluate the effect of interval between inseminations and estrus to ovulation interval (EOI) on fertility and the impact of each AI using fetal paternity in mature gilts when using FTS. Semen from 11 boars was collected and frozen in 0.5mL straws at 1.4×10(9)cells/mL. Time of estrus was synchronized in gilts (n=191) using Matrix(®) and assigned at estrus to AI treatment: (1) AI at a 4h interval (34 and 38h); (2) AI at an 8h interval (30 and 38h); or (3) AI at a 16h interval (22 and 38h). Ultrasonography was performed to determine time of ovulation. In each treatment, the first and second AI were from unique sires to allow for paternity identification using microsatellite markers. Each sire was represented across first and second inseminations. Gilts received an AI with 2×10(9) motile FTS in 80mL of extender. Pregnancy rate and litter size were assessed at slaughter on Day 33 following AI. Interval between inseminations affected pregnancy rate (P<0.01) with AI intervals of 8 (80%) and 16h (75%) greater than the 4h interval (55%). The EOI also affected pregnancy rate (P<0.001) with increased pregnancy rate for gilts having ovulations by 36h (85%) compared to those having ovulations by 24 (55%) and 60h (47%) but did not differ from those having ovulations at 48h (74%). Total fetuses (10.7), number of normal fetuses (10.2) and embryo survival (70.4%) were not affected by interval between inseminations or EOI (P>0.05). There was an interaction of treatment with EOI (P<0.05). Gilts inseminated at 8 or 16h intervals and with 36 or 48h EOI showed changes (P<0.05) in the proportion of offspring sired by the second AI while gilts inseminated at a 4h interval did not. The results of this study indicate that intervals between inseminations and time of ovulation each affects pregnancy rates but not litter size, but interacts to affect the proportion of offspring sired by the second insemination. This study suggests no detrimental effect of reduced amount of time between inseminations on the fertility of sperm used in a second insemination. For practical application, in a twice daily estrous detection and double AI, the greatest pregnancy rates occur when inseminations are spaced at 8 or 16h intervals to compensate for variation in ovulation.


PREVALENCE OF DICROCOELIUM DENDRITICUM INFECTION IN SHEEP AT TAIF PROVINCE, WEST SAUDI ARABIA.
Dicrocoelium dendriticum is a common zoonotic parasite of sheep in many Regions of Saudi Arabia. In chronic infections, this parasite causes biliary cirrhosis in livers of cattle, sheep, or goats and leads to economic losses. This study compared the prevalence of Dicrocoelium dendriticum in local sheep and imported sheep in Taif Province, West Saudi Arabia. This cross-sectional survey was carried out in slaughterhouses in Taif Province. A total 2230 local sheep and 21383 imported sheep were studied. The number of injury in imported sheep was observed as 99 representing 0.46% of the number of animals examined, while no record of any injury was observed in local sheep.


Chlamydia pneumoniae serology is associated with thrombosis-related but not with plaque-related microembolization during carotid endarterectomy.
Chlamydia pneumoniae has repeatedly been associated with atherosclerotic disease. Our study was designed to clarify whether this association is based on C pneumoniae-induced transformation of a stable into an unstable atherosclerotic plaque or on stimulation of hypercoagulability leading to increased thrombotic arterial occlusions by C pneumoniae infection. Transcranial Doppler ultrasonographic monitoring of the middle cerebral artery during carotid endarterectomy offers the opportunity to study, before removal of the plaque, atherothrombotic emboli dislodging from an unstable carotid plaque (plaque-related emboli) and emboli related to (excessive) thrombus formation at the endarterectomy site after removal of the plaque and restoration of flow (thrombosis-related emboli). C pneumoniae IgA (> or =1/16) and IgG (> or =1/64) seropositivity was assessed in 53 patients with symptomatic carotid artery disease undergoing carotid endarterectomy. The removed carotid plaques were studied histologically to assess plaque instability. Plaque- and thrombosis-related emboli were registered in 43 patients with an adequate transtemporal window. IgA seropositivity (58%) was associated significantly with thrombosis-related embolization (P=0.030) but not with plaque-related embolization or with histological plaque instability. C pneumoniae serology is associated with microembolization after endarterectomy and restoration of flow. Since these microemboli represent platelet aggregations and are related to cerebrovascular complications, our data suggest that C pneumoniae infection contributes to cerebrovascular events in patients with carotid artery disease through stimulation of thrombosis.


Exosomes from NSC-34 Cells Transfected with hSOD1-G93A Are Enriched in miR-124 and Drive Alterations in Microglia Phenotype.
Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurodegenerative disorder affecting motor neurons (MNs). Evidences indicate that ALS is a non-cell autonomous disease in which glial cells participate in both disease onset and progression. Exosomal transfer of mutant copper-zinc superoxide dismutase 1 (mSOD1) from cell-to-cell was suggested to contribute to disease dissemination. Data from our group and others showed that exosomes from activated cells contain inflammatory-related microRNAs (inflamma-miRNAs) that recapitulate the donor cell. While glia-derived exosomes and their effects in neurons have been addressed by several studies, only a few investigated the influence of motor neuron (MN)-derived exosomes in other cell function, the aim of the present study. We assessed a set of inflamma-miRs in NSC-34 MN-like cells transfected with mutant SOD1(G93A) and extended the study into their derived exosomes (mSOD1 exosomes). Then, the effects produced by mSOD1 exosomes in the activation and polarization of the recipient N9 microglial cells were investigated. Exosomes in coculture with N9 microglia and NSC-34 cells [either transfected with either wild-type (wt) human SOD1 or mutant SOD1(G93A)] showed to be transferred into N9 cells. Increased miR-124 expression was found in mSOD1 NSC-34 cells and in their derived exosomes. Incubation of mSOD1 exosomes with N9 cells determined a sustained 50% reduction in the cell phagocytic ability. It also caused a persistent NF-kB activation and an acute generation of NO, MMP-2, and MMP-9 activation, as well as upregulation of IL-1β, TNF-α, MHC-II, and iNOS gene expression, suggestive of induced M1 polarization. Marked elevation of IL-10, Arginase 1, TREM2, RAGE, and TLR4 mRNA levels, together with increased miR-124, miR-146a, and miR-155, at 24 h incubation, suggest the switch to mixed M1 and M2 subpopulations in the exosome-treated N9 microglial cells. Exosomes from mSOD1 NSC-34 MNs also enhanced the number of senescent-like positive N9 cells. Data suggest that miR-124 is translocated from the mSOD1 MNs to exosomes, which determine early and late phenotypic alterations in the recipient N9-microglial cells. In conclusion, modulation of the inflammatory-associated miR-124, in mSOD1 NSC-34 MNs, with potential benefits in the cargo of their exosomes may reveal a promising therapeutic strategy in halting microglia activation and associated effects in MN degeneration.


Age, skill transfer, and conjunction search.
Ten younger and ten older adults were provided with 16 sessions of conjunction search under consistent mapping, where target and distractors remain constant in identity. The target and one distractor were reversed after every fourth session. After the first four training sessions, on target-present trials, display size slopes were near zero for both age groups. However, on target-absent trials, older adults continued to show significantly larger display size effects than younger adults. There were no systematic age differences in either the probability of fixating objects that possess the target's features or in the amount of disruption at any reversal. Thus, although older adults exhibited more conservative criteria in visual search, they developed proficient and flexible search skill to the same degree as their younger counterparts. These data have implications for models of visual attention, skill acquisition, and cognitive aging.


A Prospective Study Comparing Quality of Life after Laparoscopic Donor Nephrectomy versus Open Donor Nephrectomy.
Laparoscopic donor nephrectomy (LDN) has advantages over open donor nephrectomy (ODN), with less bleeding and pain, and earlier discharge. However, the quality of life (QOL) has not been compared between these techniques. All consecutive donors undergoing left LDN or ODN from 2013 to 2015 at our center were included. The donors' QOL was measured with the brief World Health Organization QOL (WHOQOL-BREF) and the Hospital Anxiety and Depression Scale (HADS) questionnaires preoperatively and at 3 and 6 months postoperatively. Cosmesis was measured by the body image questionnaire, and patient satisfaction was scored on a verbal rating scale at 3 and 6 months. Of the 264 donors, 228 met the inclusion criteria (100 - LDN and 128 - ODN). The LDN group showed no difference in WHOQOL-BREF scores at 3 months, and significant improvements in the psychological and social domains at 6 months versus the baseline. The ODN group showed a significant decrease in the physical, psychological, and environmental domains at 3 and 6 months versus the baseline. Compared with the ODN group, the LDN group had better QOL scores at 3 and 6 months in all domains. The responses to the HADS questionnaire were similar between the groups at all time points. The mean body image, cosmesis, and satisfaction scores were significantly better in the LDN group. LDN minimized the negative effects of surgery on the donors' QOL, with improvements in the long-term psychological and social aspects versus the baseline. LDN was more cosmetic and was associated with higher satisfaction than ODN.


An out-patient review of facial palsy in the community.
The true psychological morbidity of facial palsy, including Bell's palsy, is not fully understood. This study addresses this problem by a voluntary questionnaire of ENT out-patients. It is not a representative study but indicates that the long-term sequelae of facial palsy is under-estimated by clinicians.


Influence of FIS on the transcription from closely spaced and non-overlapping divergent promoters for an aminoacyl-tRNA synthetase gene (gltX) and a tRNA operon (valU) in Escherichia coli.
The gltX gene, encoding the glutamyl-tRNA synthetase (GluRS), and the valU operon, whose transcripts contain three tRNAVal/UAC and one tRNALys/UUU, are adjacent and divergently transcribed. It is the only known case of adjacent genes encoding an aminoacyl-tRNA synthetase and a tRNA precursor in Escherichia coli. The gltX promoters (P1, P2 and P3) direct the synthesis of transcripts non-overlapping with and divergent from the one initiated at the valU promoter. We report that their promoter region (250 bp) contains three binding sites for the factor for inversion stimulation (FIS), centred at positions -71, -91 and -112 from the valU transcription initiation site, and that the destruction of any of these sites does not prevent the binding of FIS to the others. As FIS is one of the major positive regulators of stable RNA operons, we have studied its role on gltX and valU transcription. FIS stimulates valU transcription in vitro and about twofold in vivo during steady-state exponential growth. In contrast, gltX transcription is repressed by the presence of FIS in vitro and about twofold in vivo during growth acceleration when a decrease in GluRS concentration was observed. Under all conditions tested, most of the gltX transcripts start at the P3 promoter. Nested deletions of this regulatory region reveal that the FIS-dependent repression of the gltX-P3 promoter is abolished after the removal of the valU promoter, and is not altered by the additional removal of the FIS binding sites; moreover, in vivo transcription from gltX-P1 and/or gltX-P2 present on some of these regulatory region variants is modulated by the nature of the upstream region by FIS and is sometimes stronger than that from gltX-P3. These results show that the strength and the site of gltX transcription initiation are influenced by the upstream region up to and including the valU promoter; furthermore, they indicate that although these adjacent genes are involved in the first step of protein biosynthesis and share cis and trans regulatory elements, their transcription is non-co-ordinate.


Studying Molecular Interactions at the Single Bond Level with a Laminar Flow Chamber.
During the last decade, many investigators developed new methodologies allowing to study ligand-receptor interactions with unprecedented accuracy, up to the single bond level. Reported results include information on bond mechanical properties, association behaviour of surface-attached molecules, and dissection of energy landscapes and reaction pathways. The purpose of the present review is to discuss the potential and limitations of laminar flow chambers operated at low shear rates. This includes a brief review of basic principles, practical tips and problems associated with data interpretation. It is concluded that flow chambers are ideally suited to analyze weak interactions between a number of biomolecules, including the main families of adhesion receptors such as selectins, integrins, cadherins and members of the immunoglobulin superfamily. The sensitivity of the method is limited by the quality of surfaces and efficiency of the studied ligand-receptor couple rather than the hardware. Analyzing interactions with a resolution of a piconewton and a few milliseconds shows that ligand-receptor complexes may experience a number of intermediate binding states, making it necessary to examine the definition of association and dissociation rates. Finally, it is emphasized that association rates measured on surface-bound molecules are highly dependent on parameters unrelated to binding surfaces.


Influence of process (extrusion/thermo-compression, casting) and lentil protein content on physicochemical properties of starch films.
Starch films often present high water sensitivity, affecting their barrier and mechanical properties. The effects of processing technique, extrusion/thermo-compression and casting, and lentil protein concentration (0, 0.75 wt.% and 1.5 wt.%) on biodegradable starch films were investigated. Extrusion/thermo-compression process increased in 90% the mechanical resistance of starch films produced following the casting methodology and decreased their moisture content, water solubility and water vapor permeability in 35%, 23%, and 50%, respectively. In the presence of the protein, the mechanical properties (Young modulus and stress at break) and the water tolerance improved due to the crosslinking phenomenon prompted between the protein and the polymeric backbone, being these effects more pronounced on the extruded formulations. All samples resulted thermal stable until 240 °C and biodegraded in compost in 5 weeks. This work revealed that extrusion/thermo-compression process and proteins as crosslinking of starch are two alternatives to improve the drawbacks of starch-based materials.


Intermolecular vibrational spectra of C3v CXY3 Molecular Liquids, CHCl3, CHBr3, CFBr3, and CBrCl3.
We report the quality anisotropic intermolecular vibrational spectra within the frequency range 0.5-800 cm(-1) of four C(3v) CXY(3) molecular liquids, CHCl(3), CHBr(3), CFBr(3), and CBrCl(3), by means of femtosecond optical-heterodyne-detected Raman-induced Kerr effect spectroscopy. The results show that the first moment of the intermolecular vibrational spectrum is proportional to the square root of the value of the surface tension divided by the liquid density. This implies that the intermolecular vibrational spectrum reflects the bulk properties of the liquids. To understand the molecular-level aspects of the intermolecular vibrational spectra of the liquids, the spectra are compared with the molecular properties such as molecular weight, rotational constants, and bimolecular interaction energy. Overall, the first moment of the spectrum moderately correlates to the inverse square roots of both the molecular weight and the fast rotational constant. Therefore, the molecular properties are responsible for the intermolecular vibrational spectrum. Plots of the first moment of the intermolecular vibrational spectrum vs the square root of the value of the simple bimolecular interaction energy divided by the molecular surface area and the molecular weight show a linear correlation in the case of the oblate symmetric top molecular liquids, CHCl(3), CHBr(3), and CFBr(3). However, CBrCl(3), which is a prolate symmetric top molecular liquid, does not show the same correlation for the oblate molecular liquids.


Cofactor-independent RNA editing by a synthetic S-type PPR protein.
Pentatricopeptide repeat (PPR) proteins are RNA-binding proteins that are attractive tools for RNA processing in synthetic biology applications given their modular structure and ease of design. Several distinct types of motifs have been described from natural PPR proteins, but almost all work so far with synthetic PPR proteins has focused on the most widespread P-type motifs. We have investigated synthetic PPR proteins based on tandem repeats of the more compact S-type PPR motif found in plant organellar RNA editing factors and particularly prevalent in the lycophyte Selaginella. With the aid of a novel plate-based screening method, we show that synthetic S-type PPR proteins are easy to design and bind with high affinity and specificity and are functional in a wide range of pH, salt and temperature conditions. We find that they outperform a synthetic P-type PPR scaffold in many situations. We designed an S-type editing factor to edit an RNA target in E. coli and demonstrate that it edits effectively without requiring any additional cofactors to be added to the system. These qualities make S-type PPR scaffolds ideal for developing new RNA processing tools.


The development of a scale to assess war-time atrocities: the War Events Scale.
The War Events Scale (WES) was developed to assist clinicians with the assessment of war zone veterans' exposure to, and participation in war-time atrocities distinct from combat, and their current distress from these events. Data concerning content validity, test-retest reliability, internal consistency, as well as correlational data with the Combat Exposure Scale and the Mississippi Scale are presented. Results indicate that the WES does have adequate internal consistency and test-retest reliability, and correlates moderately with the Combat Exposure and Mississippi Scales. Initial results suggest that the WES may be helpful in collecting extremely sensitive information concerning the exposure of war zone veterans to atrocities.


Medication Use and Costs Among Older Adults Aged 90 Years and Older in Italy.
Older adults are often affected by multiple chronic conditions and experience geriatric syndromes that may affect the risk/benefit profile of medications. Little is known about the use of such medications in the older population. This article describes medication use and costs in Italian adults aged ≥90 years. Data from the 2019 Pharmaceutical Prescriptions database, concerning data on medications reimbursed by the Italian National Health Service, were analyzed in terms of prevalence and amount of use expressed as defined daily dose/1,000 users (DDD/1,000 users/day), accounting for different age-groups and sex. All individuals aged ≥90 years used at least one medication, with a mean number of 3128 DDD/1,000 users/day corresponding to an annual cost of 683 euros per user. Both use and costs linearly decreased with increasing age, with men accounting for a higher amount of DDD/1,000 users and costs than women across all age-groups. Antihypertensives (1330 DDD/1,000 inhabitants), antiplatelet agents (337 DDD/1,000 inhabitants), medications for peptic ulcer and gastroesophageal reflux (328 DDD/1,000 inhabitants), and lipid-lowering agents (166 DDD/1,000 inhabitants) were the most frequently used medications. We observed a progressive decrease in the usage of the majority of medications with increasing age, with the exception of antibiotics and antipsychotics. Individuals aged ≥90 years used a lower DDD/1,000 users, with an associated decrease in annual costs. The persistent use of preventive medications highlights the potential lack of awareness regarding medication rationalization and guidance for optimizing prescriptions. Our findings highlight the need for further initiatives to improve medications' appropriateness in these older age-groups.


Morphological and physicochemical evaluation of the propranolol HCl-Eudragit® RS100 electrosprayed nanoformulations.
The aim of this study was to fabricate propranolol hydrochloride (Prop. HCl) (as a water-soluble drug):Eudragit® RS100 (Eud) nanobeads and nanofibres applying the electrospraying method as an economical and one-step technique. Different ratios of Prop. HCl:Eud (i.e. 1:5 and 1:10) at total solution concentrations of 10-20% W/V were investigated. The FE-SEM studies revealed that morphology and size of the samples were highly affected by the solution concentration; so that, the nanobeads (a mean diameter of 82.9 nm) were formed in low concentration and at the highest concentration of the solution, nanofibres (a mean diameter of 232.3 nm) were resulted. Besides the morphological changes, the size of processed nanoformulations was increased with an increment of the solution concentrations. X-ray diffraction results as well as DSC thermograms clearly indicated that the drug crystallinity decreased in the electrosprayed samples. Furthermore, in vitro dissolution test showed that the electrosprayed samples had relatively slower release patterns toward the pure drug and physical mixtures, where the samples with the drug:polymer ratio of 1:10 indicated a faster release rate toward 1:5 ratio; nevertheless, the concentration of the injected formulations did not remarkably impressed the release behaviours. The current study established the suitability of electrospraying method in the fabrication of the water-soluble drugs nanobeads/nanofibres; however, in vivo effectiveness of the prepared nanoformulations should be meticulously considered.


Pharmacokinetics, safety, and tolerability of ceftolozane/tazobactam in healthy Japanese, Chinese, and white subjects.
Ceftolozane/tazobactam, a novel antibacterial with potent activity against Gram-negative pathogens, was developed for treatment of complicated urinary tract infections, including pyelonephritis, and intra-abdominal infections. A phase 1 pharmacokinetic (PK) study of ceftolozane/tazobactam in healthy Japanese, Chinese, and white volunteers was conducted to assess the potential effect of ethnicity on PK. The PK of ceftolozane, tazobactam, and tazobactam metabolite M1 was compared after single 1.5- and 3-g intravenous doses of ceftolozane/tazobactam. Ten Japanese, nine Chinese, and ten white subjects were enrolled, and 27 completed all doses of study medication. Dose-normalized PK parameters for ceftolozane and tazobactam were similar among Japanese, Chinese, and white subjects (at 1.5-g and 3-g doses, ceftolozane area under the plasma concentration-time curve from zero to infinity [AUC0-∞ ] = 166.3, 165.9, and 185.5 h μg/mL, respectively, and 157.7, 158.5, and 181.2 h μg/mL, respectively; tazobactam AUC0-∞ = 48.5, 43.2, and 50.1 h μg/mL, respectively, and 47.3, 43.7, and 50.0 h μg/mL, respectively. The 90% CIs of their ratio estimates were within the range 0.80 to 1.25 with the exception of AUC0-∞ for ceftolozane after the 3-g dose (0.79). The cumulative amount of ceftolozane and tazobactam excreted in urine was similar among ethnic groups. For all groups, treatment-emergent adverse events (AEs) were mild; no deaths or serious AEs were reported. The PK of ceftolozane/tazobactam was approximately dose proportional (i.e. doubling the dose approximately doubles the exposure) and similar among the groups. No dosage adjustment is needed for ceftolozane/tazobactam in Japanese and Chinese patients.