Divergent roles of IFNs in the sensitization to endotoxin shock by lactate dehydrogenase-elevating virus.
The effect of mouse infection with lactate dehydrogenase-elevating virus (LDV), a usually non-pathogenic virus, on concomitant bacterial endotoxin shock was analyzed, in terms of lethality and cytokine production. A strong enhancement of susceptibility to the shock was observed in mice acutely infected with this virus. It correlated with a sharp increase of tumor necrosis factor and leukemia inhibitory factor production and was controlled by the mouse genetic background. The viral infection led to an imbalance in the cytokine response to LPS, with an enhancement of pro-inflammatory cytokines, including IL-18 and IFN-gamma and a delayed secretion of anti-inflammatory IL-10 that could result in exacerbated macrophage activation. Enhanced IFN-gamma production was involved in the virus-induced susceptibility to shock. In sharp contrast with other viral infections, IFN-alpha/beta diminished IFN-gamma production and the resulting increased response to LPS in LDV-infected animals.


Comparison of methohexital and propofol for electroconvulsive therapy: effects on hemodynamic responses and seizure duration.
Electroconvulsive therapy can produce severe disturbances in the cardiovascular system, most commonly a transient period of hypertension. This study was designed to determine whether propofol, in comparison with methohexital, would attenuate this hypertensive response. Fifteen patients were studied during courses of six ECT administrations, each patient receiving propofol or methohexital on different occasions. Arterial pressure, heart rate, and cardiac rhythm were recorded. The induction doses were 1.08 +/- 0.03 mg.kg-1 of methohexital, and 1.60 +/- 0.04 mg.kg-1 of propofol. Systolic pressure, diastolic pressure, and heart rate were consistently lower following propofol than methohexital (P less than 0.005). The mean maximum increase over baseline systolic pressure was 2.1 +/- 2.9 mmHg with propofol, and 26.7 +/- 4.5 mmHg with methohexital (P less than 0.001). Cardiac rhythm abnormalities were infrequent, and their incidence did not differ significantly between the two induction agents (P greater than 0.3). The duration of seizures, as measured clinically, was reduced with propofol (17.9 +/- 2.5 s) in comparison with methohexital (30.9 +/- 2.8 s) (P less than 0.001). Recovery times were similar for the two agents. Since the role of seizure duration in the therapeutic efficacy of ECT remains controversial, propofol may be a useful induction agent for this procedure.


Proteomic analysis of proteins differentially expressed in preeclamptic trophoblasts.
To identify differential trophoblastic proteins associated with preeclampsia (PE) by proteomic analysis. We isolated and purified placental trophoblasts from normotensive pregnant women and patients with PE by a continuous Percoll gradient. The expression of proteins was determined by sliver staining after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Proteins of interest were identified using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The overall trophoblastic protein expression patterns in preeclamptic and corresponding normotensive placentas were quite similar except for some areas. Of 34 differentially expressed protein spots (p < 0.05 by paired t-test), seven differential proteins from nine spots were identified by MALDI-TOF-MS. The expression of the following proteins was repressed (p < 0.01): disulfide isomerase ER-60, peroxiredoxin 2, and Delta3,5-Delta2,4-dienoyl-CoA isomerase. Four proteins (protein disulfide isomerase precursor, endoplasmic reticulum resident protein, dihydrolipoyl dehydrogenase and TIM21-like protein) were found to be significantly upregulated in PE (p < 0.01). We identified several proteins with significant altered expression in PE using 2D-PAGE. This method is a powerful technique with which to search for not only quantitative but also qualitative changes in a biological process of interest.


Intermittent hypoxia differentially affects metabolic and oxidative stress responses in two species of cyprinid fish.
Oxygen fluctuations are common in freshwater habitats and aquaculture and can impact ecologically and economically important species of fish like cyprinids. To gain insight into the physiological responses to oxygen fluctuations in two common cyprinid species, we evaluated the impact of short-term intermittent hypoxia on oxidative stress and metabolic parameters (including levels of prooxidants and oxidative lesions, antioxidants, mitochondrial enzyme activities, mitochondrial swelling, markers of apoptosis, autophagy and cytotoxicity) in silver carp Hypophthalmichthys molitrix and gibel carp Carassius gibelio. During hypoxia, gibel carp showed higher baseline levels of antioxidants and less pronounced changes in oxidative and metabolic biomarkers in the tissues than silver carp. Reoxygenation led to a strong shift in metabolic and redox-related parameters and tissue damage, indicating high cost of post-hypoxic recovery in both species. Species-specific differences were more strongly associated with oxidative stress status, whereas metabolic indices and nitrosative stress parameters were more relevant to the response to hypoxia-reoxygenation. Overall, regulation of energy metabolism appears more critical than the regulation of antioxidants in the response to oxygen deprivation in the studied species. Further research is needed to establish whether prioritizing metabolic over redox regulation during hypoxia-reoxygenation stress is common in freshwater cyprinids.


No. 223-Content of a Complete Routine Second Trimester Obstetrical Ultrasound Examination and Report.
To review the benefits of and requirements for a complete second trimester ultrasound and the documentation needed. A complete second trimester ultrasound provides information about the number of fetuses, the gestational age, the location of the placenta, and fetal and maternal anatomy. In the production of this document, the American Institute of Ultrasound in Medicine's "Practice Guideline for the Performance of Obstetric Ultrasound Examinations," the American College of Obstetricians and Gynecologists' practice bulletin, "Ultrasound in Pregnancy," and the Royal College of Obstetricians and Gynaecologists' Working Party Report, "Ultrasound Screening" were reviewed. PubMed and the Cochrane Database were searched using the words "routine second trimester obstetrical ultrasound." The evidence was evaluated using the guidelines developed by the Canadian Task Force on Preventive Health Care. A routine complete second trimester ultrasound between 18 and 22 weeks and a complete ultrasound report will provide the best opportunity to diagnose fetal anomalies and to assist in the management of prenatal care. It will also reduce the number of ultrasound examinations done during the second trimester for completion of fetal anatomy survey. The costs are those involved with the performance of obstetrical ultrasound. This is a revision of previous guidelines; information from other consensus reviews from medical publications has been used. The Society of Obstetricians and Gynaecologists of Canada.


Apoptotic caspases suppress mtDNA-induced STING-mediated type I IFN production.
Activated caspases are a hallmark of apoptosis induced by the intrinsic pathway, but they are dispensable for cell death and the apoptotic clearance of cells in vivo. This has led to the suggestion that caspases are activated not just to kill but to prevent dying cells from triggering a host immune response. Here, we show that the caspase cascade suppresses type I interferon (IFN) production by cells undergoing Bak/Bax-mediated apoptosis. Bak and Bax trigger the release of mitochondrial DNA. This is recognized by the cGAS/STING-dependent DNA sensing pathway, which initiates IFN production. Activated caspases attenuate this response. Pharmacological caspase inhibition or genetic deletion of caspase-9, Apaf-1, or caspase-3/7 causes dying cells to secrete IFN-β. In vivo, this precipitates an elevation in IFN-β levels and consequent hematopoietic stem cell dysfunction, which is corrected by loss of Bak and Bax. Thus, the apoptotic caspase cascade functions to render mitochondrial apoptosis immunologically silent.


Thermoluminescent dosimetry of the selectseed 125I interstitial brachytherapy seed.
This work presents experimental dosimetry results for the new selectSeed 125I prostate seed design for use with the seedSelectron afterloading device, in accordance with the AAPM advisory that all new low energy interstitial brachytherapy seeds should undergo one Monte Carlo (MC) and at least one experimental dosimetry characterization. TLD dosimetry was performed using 120 cylindrical LiF TLD type-100 rods calibrated using a 6 MV photon beam. They were irradiated in solid water phantoms for the experimental determination of the seed dose rate constant, radial dose functions and anisotropy functions. MC simulations were performed for the determination of the TLDs relative energy response that was found position independent and equal to 1.40+/-0.03, and for the calculation of the ratio of dose in liquid water to dose in solid water that was found to be well described by Dliquidwater/Dsolidwater= 1.013*r+0.030 presenting only a minor dependence on polar angle. The selectSeed dose rate constant in liquid water was found equal to 0.938+/-0.065 cGy h(-1) U(-1), which agrees within experimental uncertainties with corresponding MC results of lambdaselect Seed=0.954+/-0.005 cGy h(-1) U(-1). The experimental radial dose and anisotropy function results were also found in good agreement with corresponding MC calculations.


Laboratory measurements of the recombination of PAH ions with electrons: implications for the PAH charge state in interstellar clouds.
Laboratory measurements of the recombination of polycyclic aromatic hydrocarbon (PAH) ions with electrons are presented. Experimental data have been obtained at room temperature for azulene (C10H8) and acenaphthene (C12H10) cations by the Flowing Afterglow with PhotoIons method. The results confirm that the recombination of PAH ions is fast although well below the geometrical limit. The set of our recent and present measurements reveal a definite trend of increasing rate with the number of carbon atoms of the PAH. This behaviour that needs further characterization is potentially of great interest for charge state models as recombination is a dominant mechanism of PAH ion destruction in the interstellar medium. The design of experiments to measure the recombination of larger PAHs and their temperature dependence is discussed.


Defective P2Y purinergic receptor function: A possible novel mechanism for impaired glucose transport.
Extracellular ATP is an ubiquitous mediator that regulates several cellular functions via specific P2 plasma membrane receptors (P2Rs), for which a role in modulating intracellular glucose metabolism has been recently suggested. We have investigated glucose uptake in response to P2Rs stimulation in fibroblasts from type 2 diabetic (T2D) patients and control subjects. P2Rs expression was evaluated by RT-PCR; intracellular calcium release by fluorometry; glucose transporter (GLUT1) translocation by immunoblotting and chemiluminescence; glucose uptake was measured with 2-deoxy-D-[1-(3)H]glucose (2-DOG) and ATP by luminometry. Cells from T2D patients, in contrast to those from healthy controls, showed no increase in glucose uptake after ATP stimulation; extracellular ATP caused, however, a similar GLUT1 recruitment to the plasma membrane in both groups. P2Rs expression did not differ between fibroblasts from diabetic and healthy subjects, but while plasma membrane depolarization, a P2X-mediated response was similar in both groups, no evident intracellular calcium increase was detectable in the cells from the former group. The calcium response in fibroblasts from diabetics was restored by co-incubation with apyrase or hexokinase, suggesting that P2YRs in those cells were normally expressed but chronically desensitised. In support to this finding, fibroblasts from T2D subjects secreted a two-fold larger amount of ATP compared to controls. Pre-treatment with apyrase or hexokinase also restored ATP stimulated glucose uptake in fibroblasts from diabetic subjects. These results suggest that extracellular ATP plays a role in the modulation of glucose transport via GLUT1, and that the P2Y-dependent GLUT1 activation is deficient in fibroblasts from T2D individuals. Our observations may point to additional therapeutic targets for improving glucose utilization in diabetes.


Cancer Center Website Rankings in the USA: Expanding Benchmarks and Standards for Effective Public Outreach and Education.
The 68 National Cancer Institute (NCI)-designated comprehensive and cancer centers have been tasked with leading the campaign in the fight against cancer, as well as providing education and outreach to the public. Therefore, it is important for these organizations to have an effective online presence to disseminate information and engage patients. The purpose of this study was to assess both the functionality and usability of cancer centers' websites. The 68 center web domains were evaluated using two separate but complementary approaches. First, a webcrawler was used to score each website on five dimensions: accessibility, content, marketing, technology, and usability. Rankings on each dimension and an average ranking were calculated for all 68 centers. Second, a three-reader system was used to determine a list of all functionalities present on the websites. Both webcrawler scores and functionality prevalence were compared across center type. No differences were observed in webcrawler scores between comprehensive and cancer centers. Mean scores on all dimensions ranged between 5.47 and 7.09. For the functionality assessment, 64 unique functions were determined and categorized into 12 domains, with the average center possessing less than 50 % of the functions. This census assessment of NCI centers' websites suggests the need for improvement to capitalize on new dissemination platforms available online. Progress in development of this technology can help achieve the goals of public education and outreach to a broad audience. This paper presents performance guidelines evaluated against best-demonstrated practice to facilitate social media use improvement.


Pharmacotherapy for recently evacuated military casualties.
In late 1990 during the massive Operation Desert Shield/Storm (ODS) deployment of United Nations troops, the VA/DoD Joint Contingency Plan was activated. Worst case scenarios projected tens of thousands of medical evacuees from the Persian Gulf and predicted that U.S. Military Communication Zone and CONUS bed capacity would quickly be saturated. As a result, a massive educational program was rapidly implemented to prepare 80 VA hospitals to receive ODS personnel within 1-3 weeks of their evacuation from battalion aid stations or other front-echelon medical facilities. This article was written as part of this educational effort. Noting that there were no published articles on pharmacotherapy for recently evacuated military casualties, the authors wrote the present manuscript for inclusion in the ODS Clinical Packet prepared by the VA's National Center for PTSD and circulated to VA mental health professionals. Since the published literature still lacks an article on this subject and since we believe that our comments on pharmacotherapy apply to survivors of most acute traumatic events, we present our observations and recommendations with the hope that they may be useful to military, VA, and civilian practitioners.


Microinjection of kynurenic acid in the rostral nucleus of the tractus solitarius disrupts spatiotemporal aspects of mechanically induced tracheobronchial cough.
The importance of neurons in the nucleus of the solitary tract (NTS) in the production of coughing was tested by microinjections of the nonspecific glutamate receptor antagonist kynurenic acid (kyn; 100 mM in artificial cerebrospinal fluid) in 15 adult spontaneously breathing anesthetized cats. Repetitive coughing was elicited by mechanical stimulation of the intrathoracic airway. Electromyograms (EMG) were recorded from inspiratory parasternal and expiratory transversus abdominis (ABD) muscles. Bilateral microinjections of kyn into the NTS rostral to obex [55 ± 4 nl total in 2 locations (n = 6) or 110 ± 4 nl total in 4 locations (n = 5)], primarily the ventrolateral subnucleus, reduced cough number and expiratory cough efforts (amplitudes of ABD EMG and maxima of esophageal pressure) compared with control. These microinjections also markedly prolonged the inspiratory phase, all cough-related EMG activation, and the total cough cycle duration as well as some other cough-related time intervals. In response to microinjections of kyn into the NTS rostral to the obex respiratory rate decreased, and there were increases in the durations of the inspiratory and postinspiratory phases and mean blood pressure. However, bilateral microinjections of kyn into the NTS caudal to obex as well as control vehicle microinjections in the NTS location rostral to obex had no effect on coughing or cardiorespiratory variables. These results are consistent with the existence of a critical component of the cough rhythmogenic circuit located in the rostral ventral and lateral NTS. Neuronal structures of the rostral NTS are significantly involved specifically in the regulation of cough magnitude and phase timing.NEW & NOTEWORTHY The nucleus of the solitary tract contains significant neuronal structures responsible for control of 1) cough excitability, 2) motor drive during cough, 3) cough phase timing, and 4) cough rhythmicity. Significant elimination of neurons in the solitary tract nucleus results in cough apraxia (incomplete and/or disordered cough pattern). The mechanism of the cough impairment is different from that for the concomitant changes in breathing.


Adaptation of a nematode parasite to living within the mammalian epithelium.
Trichuris muris is a large metazoan pathogen that has been proposed to live intracellularly within living host intestinal epithelial cells. We sought to determine how Trichuris bores its way through the mucosal epithelium and to elucidate the parasite strategies for taking advantage of this intracellular niche. Since the apical surface of the mucosal epithelium is stabilized by the actin cytoskeleton and cell junctions, it remains intact over the worm following its entry into cells. In contrast, non-stabilized lateral membranes of the host epithelial cells are ruptured and cells are killed to form an inert syncytial tunnel. The ventral surface of the nematode worm is studded by pores that overlie bacillary cells; these pores penetrate through the cuticle and are in direct contact with host cytoplasm. From scanning electron micrographs of isolated worms, we calculate that each adult contains approximately 50,000 bacillary cells. The apical surface of the bacillary cells is extensively folded into plicae 40 nm in diameter, thereby increasing the surface area many-fold. Bacillary cells lack organelles for enzyme synthesis and secretion and fail to export protons. However, by confocal light microscopy it was observed that fluorescent macromolecules in excess of 100,000 Da can penetrate into the pores. Taken together, we conclude that the bacillary cells are essential for living inside host epithelium and function predominantly in absorption of soluble molecules from the host mucosal cytoplasm, in essence behaving as an external gut epithelium that is protected from abrasion by the cuticle that surrounds the openings of the bacillary cells.


Using a PIM interface for improving computer medical image processing abilities.
Biomedical engineers in particular require fast and powerful data processing systems to process computerized tomography and magnetic resonance imaging scans and other medical imaging technologies. However, current computer data processing technologies are unable to satisfy such requirements. A promising approach to addressing these limitations is processing in memory (PIM). Unfortunately, several issues, such as the compatibility and interconnection of PIM with legacy systems, still remain. This paper proposes a standard memory bus-based PIM interface for medical image processing and a PIM platform. The proposed PIM interface can overcome problems of compatibility with legacy systems. We will adapt an embedded system based on a commercial application processor (AP) to a medical image system to verify the functions and the performance of the proposed PIM interface. Using the PIM platform, we apply the proposed PIM interface and the AP to execute an image processing program, measure the image processing times, and compare the results of the measurements. Experimental results show that while the functions of the proposed PIM interface are normal, the processing time of PIM is more than 81% faster than that of the AP. The experimental results prove that the proposed PIM interface is able to solve problems of compatibility with legacy systems. We foresee that not only the medical image processing field but also a number of academic fields and professional sectors will use PIM in their data-intensive applications.


Constrictor responses to noradrenaline, hemodynamic profile, and superoxide levels measured by hydroethidine oxidation in diabetic rats.
The aim of this study was to test the hypothesis that vascular dysfunction in neonatal streptozotocin (n-STZ)-induced diabetic rats could be associated with alterations in blood pressure, hemodynamic profile, and levels of superoxide anion. Diabetes was induced by STZ injection (160 mg/kg, i.p.) in neonate (2-d-old) Wistar rats. Using intravital microscopy the changes in mesenteric arteriolar diameters to vasoconstrictor agent noradrenaline (NA) and the levels of superoxide anion, measured by hydroethidine microfluorography, were determined in anaesthetized control and n-STZ rats. Blood pressure (BP) was determined in anaesthetized and unanaesthetized animals. Heart rate, shear rate, and blood flow velocity were also assessed. n-STZ rats showed, after 8 weeks of STZ injection, increased BP (unanaesthetized animals), hyperactivity to NA, and increased superoxide anion levels. However, heart rate, arteriolar shear rate, and blood flow velocity were unchanged in n-STZ. In conclusion, the results of the current study describe a significant increase in blood pressure, hyperactivity to NA-mediated vasoconstriction, and increased superoxide levels measured by hydroethidine oxidation. Taken together, our findings demonstrate that the compromised ability of mesenteric microvessels to respond properly in n-STZ diabetic rats is associated with several vascular alterations.


DNA-protein cross-links produced by various chemicals in cultured human lymphoma cells.
Chemicals such as cis-platinum, formaldehyde, chromate, copper, and certain arsenic compounds have been shown to produce DNA-protein cross-links in human in vitro cell systems at high doses, such as those in the cytotoxic range. Thus far there have only been a limited number of other chemicals evaluated for their ability to produce cross-links. The purpose of the work described here was to evaluate whether select industrial chemicals can form DNA-protein cross-links in human cells in vitro. We evaluated acetaldehyde, acrolein, diepoxybutane, paraformaldehyde, 2-furaldehyde, propionaldehyde, chloroacetaldehyde, sodium arsenite, and a deodorant tablet [Mega Blue; hazardous component listed as tris(hydroxymethyl)nitromethane]. Short- and long-term cytotoxicity was evaluated and used to select appropriate doses for in vitro testing. DNA-protein cross-linking was evaluated at no fewer than three doses and two cell lysate washing temperatures (45 and 65 degrees C) in Epstein-Barr virus (EBV) human Burkitt's lymphoma cells. The two washing temperatures were used to assess the heat stability of the DNA-protein cross-link, 2-Furaldehyde, acetaldehyde, and propionaldehyde produced statistically significant increases in DNA-protein cross-links at washing temperatures of 45 degrees C, but not 65 degrees C, and at or above concentrations of 5, 17.5, and 75 mM, respectively. Acrolein, diepoxybutane, paraformaldehyde, and Mega Blue produced statistically significant increases in DNA-protein cross-links washed at 45 and 65 degrees C at or above concentrations of 0.15 mM, 12.5 mM, 0.003%, and 0.1%, respectively. Sodium arsenite and chloroacetaldehyde did not produce significantly increased DNA-protein cross-links at either temperature nor at any dose tested. Excluding paraformaldehyde and 2-furaldehyde treatments, significant increases in DNA-protein cross-links were observed only at doses that resulted in complete cell death within 4 d following dosing. This work demonstrates that DNA-protein cross-links can be formed in vitro following exposure to a variety of industrial compounds and that most cross-links are formed at cytotoxic concentrations.


MiR-10a functions as a tumor suppressor in prostate cancer via targeting KDM4A.
Deregulation of microRNAs contributes to the abnormal cell growth which is frequently observed in cancer. In the current study, we detected the expression and regulatory relationship between miR-10a and Lysine-specific demethylase 4A (KDM4A) to reveal their function in prostate cancer (PCa) progression. We found that miR-10a levels were significantly decreased in PCa cell lines in comparison with the normal epithelial cell line RWPE-1. Downregulation of miR-10a levels was also observed in tumor tissues from PCa patients compared with the adjacent normal tissues. Enhanced expression of miR-10a inhibited cell proliferation and colony forming capability of PCa cells. In addition, quantitative real-time polymerase chain reaction and Western blot analysis showed a significant decrease of KDM4A in response to miR-10a elevation in PCa cells. Using dual luciferase assay, we confirmed that KDM4A was a target gene for miR-10a. Furthermore, Western blot analysis indicated that miR-10a overexpression inactivated YAP signaling and suppressed transcription of YAP target genes. Additionally, cell growth arrest and colony forming capacity inhibition induced by miR-10a overexpression could be reversed by YAP overexpression in PCa cells. More importantly, miR-10a mimics inhibited PC-3 tumor growth in nude mice accompanied with a remarkable reduction of KDM4A and YAP expression. In conclusion, our results uncovered a tumor suppressor role of miR-10a in PCa via negative regulation of KDM4A and its downstream Hippo-YAP pathway.


Inhibition of K88ab-mediated haemagglutination by polymyxin B nonapeptide.
The ability of the cyclic peptide polymyxin B nonapeptide (PMBN) to inhibit haemagglutination of erythrocytes by Escherichia coli bearing K88ab, K99 or F41 fimbriae was examined. The agent strongly inhibited K88ab-mediated haemagglutination, but had little or no effect on haemagglutination mediated by K99 or F41 fimbriae. Inhibition of K88ab-mediated haemagglutination did not result from release of fimbrial adhesins from the bacterial cell surface, nor from solubilization of K88ab receptors in erythrocytes. Since PMBN also prevented haemagglutination mediated by partially-purified K88ab fimbriae, the agent may directly obstruct access of fimbriae to their mammalian receptor binding sites.


Double-stranded RNA-independent dimerization of interferon-induced protein kinase PKR and inhibition of dimerization by the cellular P58IPK inhibitor.
The interferon (IFN)-induced, double-stranded RNA-activated protein kinase (PKR) mediates the antiviral and antiproliferative actions of IFN, in part, via its translational inhibitory properties. Previous studies have demonstrated that PKR forms dimers and that dimerization is likely to be required for activation and/or function. In the present study we used multiple approaches to examine the modulation of PKR dimerization. Deletion analysis with the lambda repressor fusion system identified a previously unrecognized site involved in PKR dimerization. This site comprised amino acids (aa) 244 to 296, which span part of the third basic region of PKR and the catalytic subdomains I and II. Using the yeast two-hybrid system and far-Western analysis, we verified the importance of this region for dimerization. Furthermore, coexpression of the 52-aa region alone inhibited the formation of full-length PKR dimers in the lambda repressor fusion and two-hybrid systems. Importantly, coexpression of aa 244 to 296 exerted a dominant-negative effect on wild-type kinase activity in a functional assay. Due to its role as a mediator of IFN-induced antiviral resistance, PKR is a target of viral and cellular inhibitors. Curiously, PKR aa 244 to 296 contain the binding site for a select group of specific inhibitors, including the cellular protein P58IPK. We demonstrated, utilizing both the yeast and lambda systems, that P58IPK, a member of the tetratricopeptide repeat protein family, can block kinase activity by preventing PKR dimerization. In contrast, a nonfunctional form of P58IPK lacking a TPR motif did not inhibit kinase activity or perturb PKR dimers. These results highlight a potential mechanism of PKR inhibition and define a novel class of PKR inhibitors. Finally, the data document the first known example of inhibition of protein kinase dimerization by a cellular protein inhibitor. On the basis of these results we propose a model for the regulation of PKR dimerization.


Recombinational instability of a chimeric plasmid in Saccharomyces cerevisiae.
Wild-type strains of Saccharomyces cerevisiae exhibit mitotic recombination between the chimeric plasmid TLC-1 and the endogenous 2mu circle that involves sequence homologies between the two plasmids that are not acted on by the 2mu circle site-specific recombination system. This generalized recombination can be detected because it separates the LEU2 and CAN1 markers of TLC-1 from each other through the formation of a plasmid containing only the S. cerevisiae LEU2 region and the 2mu circle. This derivative plasmid is maintained more stably during vegetative growth than TLC-1, and strains which carry it frequently lose the endogenous 2mu circle. Therefore, TLC-1 can provide a convenient selection for [cir0] cells. Formation of this new plasmid is greatly reduced, but not eliminated, in strains containing the rad52-1 mutation. This indicates that generalized mitotic recombination between plasmid sequences utilizes functions required for chromosomal recombination in S. cerevisiae.


Eccrine spiradenoma occurring in infancy mimicking mesenchymal tumor.
Eccrine spiradenoma (ES) rarely (less than 1%) occurs in infancy. These tumors differ from the conventional ES by the presence of superficial dermal nodules which display a less distinct two-cell pattern of immature adnexal epithelial cells and rarely ductule formation. These tumors may be mistaken for mesenchymal neoplasms involving the skin and subcutis of infants and young adults. Recognition of the histopathologic features and immunostains are required to make a definite diagnosis. We describe 2 cases of ES occurring in patients younger than one year. Detailed histopathologic and histochemical differential features of these tumors and mesenchymal neoplasms of the skin and subcutis commonly occurring in infants and young adults are discussed. The biologic behavior of infantile ES is benign, but complete excision is recommended to prevent recurrence. We speculate that these tumors may represent congenital hamartomatous growths.


Noisy spectra, long correlations, and intermittency in wave turbulence.
We study the k-space fluctuations of the wave action about its mean spectrum in the turbulence of dispersive waves. We use a minimal model based on the random phase approximation (RPA) and derive evolution equations for the arbitrary-order one-point moments of the wave intensity in the wave-number space. The first equation in this series is the familiar kinetic equation for the mean wave-action spectrum, whereas the second and higher equations describe the fluctuations about this mean spectrum. The fluctuations exhibit a nontrivial dynamics if some long coordinate-space correlations are present in the system, as it is the case in typical numerical and laboratory experiments. Without such long-range correlations, the fluctuations are trivially fixed at their Gaussian values and cannot evolve even if the wave field itself is non-Gaussian in the coordinate space. Unlike the previous approaches based on smooth initial k-space cumulants, the RPA model works even for extreme cases where the k-space fluctuations are absent or very large and intermittent. We show that any initial non-Gaussianity at small amplitudes propagates without change toward the high amplitudes at each fixed wave number. At each fixed amplitude, however, the probability distribution function becomes Gaussian at large time.


Characterization of the pharmacologic profile of a standardized soy extract in the ovariectomized rat model of menopause: effects on bone, uterus, and lipid profile.
This study was aimed to assess the effect of a standardized soy extract (SSE, Soyselect) in the ovariectomized rat model of menopause. Ovariectomized rats were treated for 6 weeks with the soy extract (50 or 100 mg/kg/day - PO), vehicle (distilled water), or 17beta-estradiol (0.5 mg/kg/day - PO). Tissue-specific estrogen agonist effects were examined using the endpoints bone mineral density, biochemical parameters of bone turnover, modulation of cytokines involved in the bone remodeling, uterine weight, uterine histology, uterine hormone receptor status, and serum lipid level. The SSE produced a bone-sparing effect associated with a slowing down in the increased bone turnover observed after ovariectomy (as indicated by measurements of serum osteocalcin levels and excretion ratio of deoxypyridinoline); changes in serum interleukin-6 levels observed after SSE suggested that this bone-sparing effect could be partly attributed to the modulation of osteoclastogenesis induced by interleukin-6. Remarkably, organ weight data and histopathologic analysis did not show any stimulatory activity of the SSE on the uterus. Immunohistochemical analysis showed a significant down-regulation of estrogen receptor-alpha (ERalpha) in uterine epithelium after 17beta-estradiol treatment, but not after treatment with the SSE; no significant differences among groups were observed in ER-alpha uterine stromal levels. After treatment with 17beta-estradiol, estrogen receptor-beta (ER-beta) expression was not modulated in the stroma or epithelium, whereas the SSE induced an up-regulation of ER-beta stromal expression. Collectively, these results suggest that the lack of stimulatory activity on the uterine epithelium using soy treatment could be due to a negligible stimulatory activity on estrogen receptor-alpha and/or to the enhanced expression observed in stromal ER-beta, the latter being considered as a negative modulator of ERalpha-mediated uterine proliferation. 17beta-estradiol, but not the SSE, down-regulated uterine epithelial progesterone receptor (PR), compared with ovariectomized rats. In the stromal compartment, progesterone receptor expression was fully up-regulated by 17beta-estradiol treatment and, to a lesser extent, by SSE treatment. The minor increase in lipid levels induced by ovariectomy was not affected by SSE administration. Finally, the lack of stimulatory activity on uterus was also confirmed in an immature female rat model. The results of this study demonstrated that the tested extract has an interesting profile of tissue-specific response, in that it is efficacious in preventing experimental osteoporosis without causing stimulation in uterus at doses that are effective in bone.


Bunching effect in single-molecule T4 lysozyme nonequilibrium conformational dynamics under enzymatic reactions.
The bunching effect, implying that conformational motion times tend to bunch in a finite and narrow time window, is observed and identified to be associated with substrate-enzyme complex formation in T4 lysozyme conformational dynamics under enzymatic reactions. Using single-molecule fluorescence spectroscopy, we have probed T4 lysozyme conformational motions under the hydrolysis reaction of polysaccharide of E. coli B cell walls by monitoring the fluorescence resonant energy transfer (FRET) between a donor-acceptor probe pair tethered to T4 lysozyme domains involving open-close hinge-bending motions. On the basis of the single-molecule spectroscopic results, molecular dynamics simulation, and a random walk model analysis, multiple intermediate states have been estimated in the evolution of T4 lysozyme enzymatic reaction active complex formation (Chen, Y.; Hu, D.; Vorpagel, E. R.; Lu, H. P. Probing single-molecule T4 lysozyme conformational dynamics by intramolecular fluorescence energy transfer. J. Phys. Chem. B 2003, 107, 7947-7956). In this Article, we report progress on the analysis of the reported experimental results, and we have identified the bunching effect of the substrate-enzyme active complex formation time in T4 lysozyme enzymatic reactions. We show that the bunching effect, a dynamic behavior observed for the catalytic hinge-bending conformational motions of T4 lysozyme, is a convoluted outcome of multiple consecutive Poisson rate processes that are defined by protein functional motions under substrate-enzyme interactions; i.e., convoluted multiple Poisson rate processes give rise to the bunching effect in the enzymatic reaction dynamics. We suggest that the bunching effect is likely common in protein conformational dynamics involved in conformation-gated protein functions.


Therapeutic Efficacy and Safety of Intense Pulsed Light for Meibomian Gland Dysfunction in Patients with Chronic Ocular Graft-Versus-Host Disease.
To evaluate the efficacy and safety of intense pulsed light combined with meibomian gland expression (IPL-MGX) for treating meibomian gland dysfunction (MGD) associated with chronic ocular graft-versus-host disease (oGVHD). This retrospective study included 18 patients (18 eyes) with Fitzpatrick skin type ≤ IV, who underwent 3 to 8 sessions of IPL-MGX. Dry eye symptomology, ocular surface parameters, and adverse events were evaluated. Of 18 eyes, 83.3% and 66.7% showed severe oGVHD and severe MGD, respectively. At 4 weeks after the final session, significant improvements in the OSDI (P < 0.001), SPEED (P = 0.001), meibum expressibility (P < 0.001), and meibum quality (P = 0.016) were observed. At 12 weeks after, the OSDI (P = 0.009), SPEED (P = 0.002), and meibum expressibility (P = 0.008) significantly improved. No adverse events owing to IPL were reported. IPL-MGX may improve the ocular symptoms, ameliorate meibomian gland secretion, and is considered as a safe treatment for MGD in oGVHD patients.


Single-neuron projectome of mouse prefrontal cortex.
Prefrontal cortex (PFC) is the cognitive center that integrates and regulates global brain activity. However, the whole-brain organization of PFC axon projections remains poorly understood. Using single-neuron reconstruction of 6,357 mouse PFC projection neurons, we identified 64 projectome-defined subtypes. Each of four previously known major cortico-cortical subnetworks was targeted by a distinct group of PFC subtypes defined by their first-order axon collaterals. Further analysis unraveled topographic rules of soma distribution within PFC, first-order collateral branch point-dependent target selection and terminal arbor distribution-dependent target subdivision. Furthermore, we obtained a high-precision hierarchical map within PFC and three distinct functionally related PFC modules, each enriched with internal recurrent connectivity. Finally, we showed that each transcriptome subtype corresponds to multiple projectome subtypes found in different PFC subregions. Thus, whole-brain single-neuron projectome analysis reveals organization principles of axon projections within and outside PFC and provides the essential basis for elucidating neuronal connectivity underlying diverse PFC functions.


Long-lasting improvement following tDCS treatment combined with a training for reading in children and adolescents with dyslexia.
Noninvasive brain stimulation transiently modulates reading ability in individuals with dyslexia by facilitating the underactive neural pathways in them. However, its long-term effects have not been determined. This study confirmed the ameliorative effects of multiple sessions of transcranial direct current stimulation (tDCS) combined with a training for reading on the reading abilities of children and adolescents with dyslexia and examined whether they are long-lasting. Twenty-six children and adolescents with dyslexia received 3 20-min sessions per week for 6 weeks (18 sessions) of left anodal/right cathodal tDCS, set to 1 mA, over the parieto-temporal regions, combined with training for reading. The participants were randomly assigned to receive active or sham treatment. Reading measures (text, high- and low-frequency words, non-words) were recorded before and immediately after the treatment and 1 and 6 months later. The long-term tolerability to tDCS was also evaluated. The active group received long-lasting benefits in reading. Specifically, the non-word reading efficiency index improved at every time point, as did the low-frequency word reading efficiency index at 1 and 6 months after the end of the treatment. No differences emerged in the sham group. No long-term adverse effects were documented. This study provides evidence of persistent improvements in reading in children and adolescents with dyslexia, constituting a new rehabilitative approach for the remediation of dyslexia.


Influence of Provider Communication on Women's Delivery Expectations and Birth Experience Appraisal: A Qualitative Study.
Although current research suggests that patient-provider prenatal communication and expectation-setting affects women's outcomes, more needs to be understood about the kinds of communication experiences that shape women's expectations, the nature of expectations that women hold, and how those expectations influence their appraisal of labor and delivery. The goal of this study is to draw connections between provider communication, birth experience expectations, and birth experience appraisals. Recently delivered mothers (n=36) were recruited at a mid-Atlantic community hospital. Using a grounded theory approach, interviews were systematically analyzed to uncover how participants perceived provider communication during their prenatal care, how participants described their expectations of the birth experience, and how expectations affected appraisals of the experience. Mothers recognize providers' use of patient-centered communication in messages of empowerment, emotional support, explanation, decision making, and elicitation. Findings posit that it is the inflexibility or flexibility of expectations that may determine mothers' appraisals of the birth experience. Mothers continue to rely on providers as partners in health care. Through patient-centered communication, providers can help mothers develop flexible expectations of the birth experience, which in turn can result in positive appraisals of delivery.


Impact of temperature on performance in two species of South African dwarf chameleon, Bradypodion pumilum and B. occidentale.
Temperature is an extrinsic factor that influences reptile behavior because of its impact on reptile physiology. Understanding the impact of temperature on performance traits is important as it may affect the ecology and fitness of ectothermic animals such as reptiles. Here, we examined the temperature dependence of performance in two species of South African dwarf chameleon (Bradypodion): one adapted to a semi-arid environment and one to a mesic environment. Ecologically relevant performance traits were tested at different temperatures to evaluate their thermal dependence, and temperature-performance breadths for 80% and 90% of each performance trait were calculated. Our results show distinct differences in the thermal dependence of speed- versus force-related performance traits. Moreover, our results show that the semi-arid species is better adapted to higher temperatures and as such has a better chance of coping with the predicted increases in environmental temperature. The mesic area-adapted species seems to be more sensitive to an increase in temperature and could therefore potentially be threatened by the predicted future climate change. However, further studies investigating the potential for acclimation in chameleons are needed to better understand how animals may respond to future climate change.


Induction of the human cathelicidin LL-37 as a novel treatment against bacterial infections.
As traditional antibiotics gradually become inefficient, there is a high demand for development of anti-infectives with a mechanism of action that is different from existing antibiotics. Current antibiotics target the pathogen directly, thereby contributing to the selection of multidrug-resistant bacterial strains. AMPs, such as the human cathelicidin LL-37, are small cationic peptides that are part of host defense. They eliminate microbes through diverse mechanisms, thereby contributing to resolution of infections and maintenance of epithelial barrier function. The multiplicity of these mechanisms of action might be a key to restrict the development of resistant bacterial strains. The discovery of LL-37-inducing components, such as butyrate and vitamin D(3), has opened new avenues to prevent or treat infections. Butyrate and vitamin D(3) are potent inducers of LL-37 but in addition, have many other effects on host immunity. Here, we summarize current data on the effects that LL-37 and its inducers display on the innate immune response and discuss the feasibility for development of these inducers as possible drugs to prevent or treat infections.


Effects of phenobarbital and diazepam on imipramine-induced changes in blood pressure, heart rate and rectal temperature of rats.
To investigate the safety of anticonvulsants in doses found equipotent in suppressing imipramine induced convulsions, the effects of diazepam (1.8 mg/kg) or phenobarbital (40 mg/kg) following a toxic dose of imipramine (50 mg/kg) on heart rate, blood pressure and body temperature were examined in male Wistar rats. Administration of imipramine alone resulted in significant decreases in blood pressure, heart rate and rectal temperature. Phenobarbital or diazepam alone failed to significantly affect any of these parameters apart from a slight reduction in rectal temperature seen with phenobarbital. Diazepam given after imipramine antagonized the imipramine-induced decrease in heart rate but increased the hypotensive and hypothermic effects. Phenobarbital failed to significantly affect the imipramine-induced changes in any of the physiological parameters studied. The present data suggests that phenobarbital may be preferable to diazepam in treatment of imipramine-induced convulsions.


Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells.
The aim of the present study was to investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI-1 cells. The transfection efficiency of fluorescein amidite (FAM)-small interfering RNA (siRNA) was determined using flow cytometry, and the protein expression levels of LPXN, phosphorylated (p)-c-Jun N-terminal kinase (JNK), p-p38 mitogen-activated protein kinase (p38 MAPK) and p-extracellular-signal-regulated kinase (ERK) were detected by western blot analysis. Proliferation was determined using the cell counting kit-8 reagent and cellular transmembrane invasion ability was determined using a Transwell chamber system. The gelatinase levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the cell culture supernatant were also analyzed by gelatin zymography. In SHI-1 cells, the optimal transfection conditions of siRNA were a cell density of 4×105 cells/ml and a ratio of siRNA/Lipofectamine® 2000 of 200 pmol/1 µl. The highest transfection efficiency of FAM-siRNA was 74.5%. In the present study, L2-siRNA was selected to effectively decrease the expression of LPXN. Following downregulation of LPXN expression by L2-siRNA, proliferation inhibition rates increased to 27.043±2.051 and cell transmembrane invasion rates decreased to 25.270±2.145 (P<0.05). The results of the western blot analysis and the gelatin zymography indicated that downregulation of LPXN expression increased the expression of p-p38 MAPK and p-JNK, and attenuated the secretion levels of MMP-2 and MMP-9. However, downregulation of LPXN expression had no effect on p-ERK expression in SHI-1 cells. The results of the present study indicated that downregulation of LPXN expression decreased the malignant proliferation and transmembrane invasion of SHI-1 cells by activating JNK and p38 MAPK, and inhibiting MMP-2 and MMP-9 secretion.


Dense doping of indium to coral-like SnO2 nanostructures through a plasma-assisted strategy for sensitive and selective detection of chlorobenzene.
A plasma-assisted strategy for densely doping indium to SnO(2) nanostructures for gas-sensing applications is reported. The morphology, structure, and composition of the as-prepared nanostructures were characterized by field emission scanning electronic microscopy (FESEM), transmission electron microscopy (TEM), x-ray diffraction (XRD), and x-ray photoelectron spectrometry (XPS), respectively. The results show that the densities of hydroxyl and carboxyl groups of the coral-like SnO(2)/carbonaceous nanocomposites are remarkably improved by using a plasma treatment (PT), which enables them to adsorb a large quantity of indium ions and thereby enhance the doping. In gas-sensing measurements, it is found that the sensor is sensitive to chlorobenzene with a high response and short response and recovery times. Besides, the gas-sensing properties of the sensor based on the In-doped SnO(2) with PT are greatly improved compared with sensors based on In-doped SnO(2) without PT and pure SnO(2). The enhanced doping and the special coral-like structure are demonstrated as the mechanism of improvement. The kinetic processes of gas adsorption and desorption are also investigated. Furthermore, it is revealed that chlorobenzene can be clearly identified from some gas references by using principal component analysis, exhibiting a good selectivity. Our findings not only provide a promising building block for developing a sensitive and selective gas sensor for environmental monitoring, but also demonstrate a novel plasma-assisted strategy which could be potentially developed as a general method for dense doping of nanomaterials.


Successful treatment of concomitant metastatic prostate cancer and B-cell non-Hodgkin's lymphoma with R-EPOCH chemotherapy regimen and antiandrogen therapy.
A 70-year-old man presented with left loin pain without urinary symptoms. Initial investigations with CT showed enlarged para-aortic, mediastinal lymph nodes, right-side renal mass and enlarged prostate. A prostatic-specific antigen (PSA) was alarmingly high at 4750 μg/L (normal <4.0 μg/L). Further investigations included positron emission tomography (PET); both prostate-specific membrane antigen and 18-fluorodeoxyglucose as well as bone scan and bone marrow examination confirmed dual malignancies with B-cell non-Hodgkin's lymphoma (B-NHL) and wide spread metastatic prostatic adenocarcinoma (PA) to the skull, spine, pelvis, liver and lungs. The patient was treated with six cycles of rituximab, etoposide, prednisone, vincristine, cyclophosphamide and doxorubicin(R-EPOCH) containing regimen for B-NHL and goserelin hormonal therapy for PA. Restaging with PET scans thereafter showed complete remission of NHL with disappearance of his metastatic PA and normalisation of PSA levels. R-EPOCH regimen and antiandrogen therapy resulted in a good outcome and remission of both malignancies.


[Rare forms of hereditary endocrine neoplasia: co-existence of pituitary adenoma and pheochromocytoma/paraganglioma].
Functioning pituitary adenomas and pheochromocytomas/paragangliomas are rare in the general population. Pituitary adenomas occur in the familial setting in approximately 5% of cases, whereas pheochromocytomas/paragangliomas can be hereditary in 30-40% of cases. Hereditary syndromes associated with pituitary adenomas include multiple endocrine neoplasia types 1 and 4, familial isolated pituitary adenomas, and Carney complex. Hereditary syndromes associated with pheochromocytomas/paragangliomas and genes, mutations in which predispose to their development, are more numerous. The first clinical descriptions of the co-occurrence of pituitary adenoma and pheochromocytoma/paraganglioma in one patient date back to the mid 20th century, however delineating such a co-occurrence into a particular syndrome («3PAs» (pituitary adenoma, pheochromocytoma, paraganglioma)) was suggested only in 2015. To date, approximately 100 cases of such a co-occurrence have been described in the literature. Mutations in genes encoding subunits of succinate dehydrogenase complex II (SDHx) are revealed in the majority of cases, much less common are mutations in MAX, MEN1 and some other genes. This review summarizes the current information on the «3PAs» syndrome.


Movement of diuron and hexazinone in clay soil and infiltrated pond water.
Pre-emergence herbicide residues were detected in domestic wells sampled near Tracy, CA. This study sought to determine the source of contamination by comparing soil distribution of diuron [N'-(3,4-dichlorophenyl)-N,N-dimethylurea] and hexazinone [3-cyclohexyl-6-(dimethylamino)-1-methyl-1,3,5-triazine-2,4(1H,3H)-dione] in an agricultural field where the soil was a cracking clay to infiltration of residues in water captured by an adjacent holding pond. Diuron and hexazinone were applied in December to a 3-yr-old alfalfa (Medicago sativa L.) crop. Water content of soil taken after major rainfall but before irrigation at 106 d after application was elevated at the lowest depth sampled centered at 953 mm, indicating water was available for percolation. Herbicide residues (reporting limit 8 microg kg(-1)) were confined above the 152 mm soil depth, even after subsequent application of two border-check surface irrigations. The pattern of distribution and concentration of residues in the soil were similar to results obtained from the LEACHM model, suggesting that macropore flow was limited to a shallow depth of soil. Herbicide residues were measured in runoff water at the first irrigation at 20 microg L(-1) for diuron and 1 microg L(-1) for hexazinone. Runoff water captured in the pond rapidly infiltrated into the subsurface soil, causing a concomitant rise in ground water elevation near the pond. Herbicide residues were also detected in the sampled ground water. We concluded that the pond was the predominant source for movement to ground water. Since addition of a surfactant to the spray mixture did not reduce concentrations in runoff water, mitigation methods will focus on minimizing infiltration of water from the pond.


Psychological interventions to improve the quality of life in Indian lung cancer patients: A neglected area.
The incidence of lung cancer is very high and evidence suggests that patients experience imbalanced emotional capabilities due to less survival rate compared to other cancers. Direct and indirect psychological interventions are mandatory to improve the outcome of lung cancer treatment. Although such interventions are being practiced in developed nations, the effects of psychological interventions on the treatment outcome in the Indian context are lacking. Since there is a definite correlation between treatment outcome and psychological issues, it is high time that clinicians in developing countries including India adopt practices to enhance the quality of life of lung cancer patients.


Eye-Tracking-Based Measurement of Social Visual Engagement Compared With Expert Clinical Diagnosis of Autism.
In the US, children with signs of autism often experience more than 1 year of delay before diagnosis and often experience longer delays if they are from racially, ethnically, or economically disadvantaged backgrounds. Most diagnoses are also received without use of standardized diagnostic instruments. To aid in early autism diagnosis, eye-tracking measurement of social visual engagement has shown potential as a performance-based biomarker. To evaluate the performance of eye-tracking measurement of social visual engagement (index test) relative to expert clinical diagnosis in young children referred to specialty autism clinics. In this study of 16- to 30-month-old children enrolled at 6 US specialty centers from April 2018 through May 2019, staff blind to clinical diagnoses used automated devices to measure eye-tracking-based social visual engagement. Expert clinical diagnoses were made using best practice standardized protocols by specialists blind to index test results. This study was completed in a 1-day protocol for each participant. Primary outcome measures were test sensitivity and specificity relative to expert clinical diagnosis. Secondary outcome measures were test correlations with expert clinical assessments of social disability, verbal ability, and nonverbal cognitive ability. Eye-tracking measurement of social visual engagement was successful in 475 (95.2%) of the 499 enrolled children (mean [SD] age, 24.1 [4.4] months; 38 [8.0%] were Asian; 37 [7.8%], Black; 352 [74.1%], White; 44 [9.3%], other; and 68 [14.3%], Hispanic). By expert clinical diagnosis, 221 children (46.5%) had autism and 254 (53.5%) did not. In all children, measurement of social visual engagement had sensitivity of 71.0% (95% CI, 64.7% to 76.6%) and specificity of 80.7% (95% CI, 75.4% to 85.1%). In the subgroup of 335 children whose autism diagnosis was certain, sensitivity was 78.0% (95% CI, 70.7% to 83.9%) and specificity was 85.4% (95% CI, 79.5% to 89.8%). Eye-tracking test results correlated with expert clinical assessments of individual levels of social disability (r = -0.75 [95% CI, -0.79 to -0.71]), verbal ability (r = 0.65 [95% CI, 0.59 to 0.70]), and nonverbal cognitive ability (r = 0.65 [95% CI, 0.59 to 0.70]). In 16- to 30-month-old children referred to specialty clinics, eye-tracking-based measurement of social visual engagement was predictive of autism diagnoses by clinical experts. Further evaluation of this test's role in early diagnosis and assessment of autism in routine specialty clinic practice is warranted. ClinicalTrials.gov Identifier: NCT03469986.


Automated Continuous Evolution of Proteins in Vivo.
We present automated continuous evolution (ACE), a platform for the hands-free directed evolution of biomolecules. ACE pairs OrthoRep, a genetic system for continuous targeted mutagenesis of user-selected genes in vivo, with eVOLVER, a scalable and automated continuous culture device for precise, multiparameter regulation of growth conditions. By implementing real-time feedback-controlled tuning of selection stringency with eVOLVER, genes of interest encoded on OrthoRep autonomously traversed multimutation adaptive pathways to reach desired functions, including drug resistance and improved enzyme activity. The durability, scalability, and speed of biomolecular evolution with ACE should be broadly applicable to protein engineering as well as prospective studies on how selection parameters and schedules shape adaptation.


Application of nucleic acid hybridization to the hepatitis delta virus research.
Cloning and sequencing of the genome of hepatitis delta virus (HDV) represented a remarkable progress in the knowledge of HDV infection and made available cDNA and RNA probes which can be successfully used to identify HDV-RNA in biological specimens. We review here the methods currently used for detecting HDV nucleic acids and report the most recent advances of the molecular biology of HDV. Finally, we examine the number of compelling reasons for the diagnostic laboratory to pursue this technology. Testing for HDV-RNA has become essential for the screening and the follow-up of both chronic HDV carriers, undergoing antiviral therapy and patients with terminal disease, eligible for liver transplant.


A Distinctive Urinary Metabolomic Fingerprint Is Linked With Endoscopic Postoperative Disease Recurrence in Crohn's Disease Patients.
Crohn's disease (CD) patients who undergo ileocolonic resection frequently have disease recurrence. The aim of this preliminary study was to identify urinary metabolomic profiles associated with disease recurrence in order to identify underlying mechanisms of recurrence and possible disease biomarkers. Biopsies from the neoterminal ileum were collected from CD patients (n = 38) after ileocolonic resection in order to assess mucosa-associated microbiota using 16S rRNA multitag pyrosequencing. Urine samples were collected, and metabolomic profiling was done using high-resolution nuclear magnetic resolution spectroscopy and a combined direct infusion liquid chromatography tandem mass spectrometry. The Rutgeerts scoring system was used to assess endoscopic postoperative recurrence of CD. There were 28 (73.7%) patients with endoscopic CD recurrence. CD patients who were in endoscopic remission had a higher abundance of Bacteroidetes and lower abundance of Fusobacteria and Proteobacteria in comparison with CD patients who had endoscopic recurrence. In addition, metabolomic profiling could also discriminate between these 2 groups of patients. Endoscopic recurrence was associated with increased concentration of urinary levoglucosan. Rutgeerts score was positively correlated with levoglucosan and propylene glycol levels. CD patients who present with endoscopic disease recurrence after surgery have a unique urinary metabolomic fingerprint that can differentiate them from CD patients who are in endoscopic remission after ileocolonic resection. In addition, mucosal-associated microbiota in CD patients with or without disease recurrence after surgery differs and correlates with some urinary metabolites.


Comparative localization of cystathionine beta synthases and cystathionine gamma lyase in canine, non-human primate and human retina.
Chronic exposure of the retina to light and high concentrations of polyunsaturated fatty acid in photoreceptor cells make this tissue susceptible to oxidative damage. As retinal degenerative diseases are associated with photoreceptor degeneration, the antioxidant activity of both hydrogen sulfide (H2S) and glutathione (GSH) may play an important role in ameliorating disease progression. H2S production is driven by cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS), the key enzymes that also drive transsulfuration pathway (TSP) necessary for GSH production. As it is currently unclear whether localized production of either H2S or GSH contributes to retinal homeostasis, we undertook a comparative analysis of CBS and CSE expression in canine, non-human primate (NHP) and human retinas to determine if these antioxidants could play a regulatory role in age-related or disease-associated retinal degeneration. Retinas from normal dogs, NHPs and humans were used for the study. Laser capture microdissection (LCM) was performed to isolate individual layers of the canine retina and analyze CBS and CSE gene expression by qRT-PCR. Immunohistochemistry and western blotting were performed for CBS and CSE labeling and protein expression in dog, NHP, and human retina, respectively. Using qRT-PCR, western blot, and immunohistochemistry (IHC), we showed that CBS and CSE are expressed in the canine, NHP, and human retina. IHC results from canine retina demonstrated increased expression levels of CBS but not CSE with post-developmental aging. IHC results also showed non-overlapping localization of both proteins with CBS presenting in rods, amacrine, horizontal, and nerve fiber cell layers while CSE was expressed by RPE, cones and Mϋller cells. Finally, we demonstrated that these enzymes localized to all three layers of canine, NHP and human retina: photoreceptors, outer plexiform layer (OPL) and notably in the ganglion cells layer/nerve fiber layer (GCL/NFL). QRT-PCR performed using RNA extracted from tissues isolated from these cell layers using laser capture microdissection (LCM) confirmed that each of CBS and CSE are expressed equally in these three layers. Together, these findings reveal that CSE and CBS are expressed in the retina, thereby supporting further studies to determine the role of H2S and these proteins in oxidative stress and apoptosis in retinal degenerative diseases.


Renal cancer staging: comparison of contrast-enhanced CT and gadolinium-enhanced fat-suppressed spin-echo and gradient-echo MR imaging.
Fifty-three consecutive patients with 61 solid or complex non-fat-containing renal masses compatible with renal cancer were examined with contrast-enhanced computed tomography (CT) and magnetic resonance (MR) imaging with pre- and postcontrast FLASH (fast low-angle shot) and fat-suppressed spin-echo sequences. CT and MR imaging were performed within a 1-month interval. CT and MR images were prospectively interpreted. Tumor detection and staging were determined in all patients. CT and MR imaging enabled detection of 54 and 58 of 61 renal tumors, respectively. CT and MR imaging showed 34 and 35 of 38 histologically proved renal tumors, respectively, in 31 patients. Tumor size on CT and MR images demonstrated good correlation and correlated well with the size of pathologic specimens of 34 of 38 resected tumors detected with CT and MR imaging (r = .99). Of the 31 tumors in 31 patients who underwent surgical resection, 24 were correctly staged with CT and 29 with MR imaging. CT and MR imaging both enabled correct staging of four of five additional tumors with biopsy proof of tumor stage. A moderate difference in staging was observed between CT and MR imaging (P = .05). CT showed 13 and MR imaging 15 of 15 tumor thrombi. CT and MR imaging both showed 11 of 11 cases of adenopathy. The results suggest that MR imaging is moderately better than CT for the detection and staging of renal cancer.


Strategy of Pseudomonas pseudoalcaligenes C70 for effective degradation of phenol and salicylate.
Phenol- and naphthalene-degrading indigenous Pseudomonas pseudoalcaligenes strain C70 has great potential for the bioremediation of polluted areas. It harbours two chromosomally located catechol meta pathways, one of which is structurally and phylogenetically very similar to the Pseudomonas sp. CF600 dmp operon and the other to the P. stutzeri AN10 nah lower operon. The key enzymes of the catechol meta pathway, catechol 2,3-dioxygenase (C23O) from strain C70, PheB and NahH, have an amino acid identity of 85%. The metabolic and regulatory phenotypes of the wild-type and the mutant strain C70ΔpheB lacking pheB were evaluated. qRT-PCR data showed that in C70, the expression of pheB- and nahH-encoded C23O was induced by phenol and salicylate, respectively. We demonstrate that strain C70 is more effective in the degradation of phenol and salicylate, especially at higher substrate concentrations, when these compounds are present as a mixture; i.e., when both pathways are expressed. Moreover, NahH is able to substitute for the deleted PheB in phenol degradation when salicylate is also present in the growth medium. The appearance of a yellow intermediate 2-hydroxymuconic semialdehyde was followed by the accumulation of catechol in salicylate-containing growth medium, and lower expression levels and specific activities of the C23O of the sal operon were detected. However, the excretion of the toxic intermediate catechol to the growth medium was avoided when the growth medium was supplemented with phenol, seemingly due to the contribution of the second meta pathway encoded by the phe genes.


Single-donor islet transplantation in type 1 diabetes: patient selection and special considerations.
Type 1 diabetes mellitus is an autoimmune disorder of the endocrine pancreas that currently affects millions of people in the United States. Although the disease can be managed with exogenous insulin administration, the ultimate cure for the condition lies in restoring a patient's ability to produce their own insulin. Islet cell allotransplantation provides a means of endogenous insulin production. Though far from perfected, islet transplants are now a proven treatment for type 1 diabetics. However, proper patient selection is critical for achieving optimal outcomes. Given the shortage of transplantable organs, selecting appropriate candidates for whom the procedure will be of greatest benefit is essential. Although many of those who receive islets do not retain insulin independence, grafts do play a significant role in preventing hypoglycemic episodes that can be quite detrimental to quality of life and potentially fatal. Additionally, islet transplant requires lifelong immunosuppression. Antibodies, both preformed and following islet infusion, may play important roles in graft outcomes. Finally, no procedure is without inherent risk and islet transfusions can have serious consequences for recipients' livers in the form of both vascular and metabolic complications. Therefore, patient-specific factors that should be taken into account before islet transplantation include aims of therapy, sensitization, and potential increased risk for hepatic and portal-venous sequelae.


A homologous mapping method for three-dimensional reconstruction of protein networks reveals disease-associated mutations.
One of the crucial steps toward understanding the associations among molecular interactions, pathways, and diseases in a cell is to investigate detailed atomic protein-protein interactions (PPIs) in the structural interactome. Despite the availability of large-scale methods for analyzing PPI networks, these methods often focused on PPI networks using genome-scale data and/or known experimental PPIs. However, these methods are unable to provide structurally resolved interaction residues and their conservations in PPI networks. Here, we reconstructed a human three-dimensional (3D) structural PPI network (hDiSNet) with the detailed atomic binding models and disease-associated mutations by enhancing our PPI families and 3D-domain interologs from 60,618 structural complexes and complete genome database with 6,352,363 protein sequences across 2274 species. hDiSNet is a scale-free network (γ = 2.05), which consists of 5177 proteins and 19,239 PPIs with 5843 mutations. These 19,239 structurally resolved PPIs not only expanded the number of PPIs compared to present structural PPI network, but also achieved higher agreement with gene ontology similarities and higher co-expression correlation than the ones of 181,868 experimental PPIs recorded in public databases. Among 5843 mutations, 1653 and 790 mutations involved in interacting domains and contacting residues, respectively, are highly related to diseases. Our hDiSNet can provide detailed atomic interactions of human disease and their associated proteins with mutations. Our results show that the disease-related mutations are often located at the contacting residues forming the hydrogen bonds or conserved in the PPI family. In addition, hDiSNet provides the insights of the FGFR (EGFR)-MAPK pathway for interpreting the mechanisms of breast cancer and ErbB signaling pathway in brain cancer. Our results demonstrate that hDiSNet can explore structural-based interactions insights for understanding the mechanisms of disease-associated proteins and their mutations. We believe that our method is useful to reconstruct structurally resolved PPI networks for interpreting structural genomics and disease associations.


Hepatitis associated with herpes viral infection in the tortoise (Testudo horsfieldii).
Herpesvirus infection in tortoises is largely characterized by the development of respiratory clinical signs. Usually lesions develop in the respiratory, oral pharyngeal, intestinal tract and are accompanied by cutaneous and ocular lesions. In chelonids affected by herpesvirus, systemic-type lesions in organs such as the liver and spleen are commonly observed. In this paper we describe a case of multifocal necrotic hepatitis associated with herpesviruses in an adult female land tortoise of the species Testudo horsfieldii. This article is the first description of a viral hepatitis in Testudo spp. with lesions compatible with herpesvirus infection, with no clinical signs or lesions in the respiratory system, oral cavity or other organs.


Lipoprotein (a), an independent cardiovascular risk marker.
Epidemiological and genetic studies have identified elevated levels of lipoprotein (a) ((Lp(a)) as a causal and independent risk factor for cardiovascular diseases (CVD). The Lp(a)-induced increased risk of CVD may be mediated by both its proatherogenic and prothrombotic mechanisms. Several guidelines recommend screening of Lp(a) level; however, there are few treatment options for the management of patients with elevated Lp(a). Several new medications for Lp(a) are under development. PCSK9 inhibitors, apolipoprotein (a)-antisense, and apolipoprotein(B-100)-antisense mipomersen have shown promising results. Lp(a) reduction will continue to be an active area of investigation.


Effectiveness of influenza vaccination: a survey within the Italian Ministry of Health personnel.
An initiative of active prevention, to promote an influenza vaccination programme, has been carried out in the Ministry of Health of Rome, Directorate General of Health Prevention: during October 2004 influenza vaccine has been actively offered to all the employees; 46 out of the 129 workers (about 36% of the total) accepted to be vaccinated. Afterward, the propensity to get influenza of the vaccinated has been compared with that of a control group, operating in the same setting, but not vaccinated. Taking into account that the groups in question is far too small, the statistical results showed that influenza vaccination of healthy working adults did not reduce the rates of influenza-like-illness (ILI), in addition, the difference between average duration of disease in unvaccinated persons was not statistically significant.


Effect of Cold Application on Pain and Bruising in Patients With Subcutaneous Injection of Low-Molecular-Weight Heparin: A Meta-Analysis.
To evaluate the effect of cold application on pain and bruising after the subcutaneous injection of low-molecular-weight heparin, 8 electronic databases were searched for randomized controlled trials and quasiexperimental studies from the inception of the databases to June 2019. Review Manager 5.3 software was used for the heterogeneity test and meta-analysis. A total of 8 studies including 694 participants were analyzed. The cold application group assessed with the Verbal Descriptor Scale pain assessment tool showed significant reductions in pain intensity immediately after injection. Compared to the control group, the cold application group showed a reduction in the occurrence of bruises at 12 hours, 24 hours, and 48 hours after injection. There was no significant difference in the area of bruising in the cold application group at 48 hours after injection, but the area of bruising at 72 hours after injection was significantly reduced. These results show that cold application can reduce the incidence of pain and bruising after subcutaneous injection of low-molecular-weight heparin and reduce the area of bruising 72 hours after injection. Additional studies with larger sample sizes are needed to confirm these findings.


The role of spatial heterogeneity in the evolution of local and global infections of viruses.
Viruses have two modes spread in a host body, one is to release infectious particles from infected cells (global infection) and the other is to infect directly from an infected cell to an adjacent cell (local infection). Since the mode of spread affects the evolution of life history traits, such as virulence, it is important to reveal what level of global and local infection is selected. Previous studies of the evolution of global and local infection have paid little attention to its dependency on the measures of spatial configuration. Here we show the evolutionarily stable proportion of global and local infection, and how it depends on the distribution of target cells. Using an epidemic model on a regular lattice, we consider the infection dynamics by pair approximation and check the evolutionarily stable strategy. We also conduct the Monte-Carlo simulation to observe evolutionary dynamics. We show that a higher local infection is selected as target cells become clustered. Surprisingly, the selected strategy depends not only on the degree of clustering but also the abundance of target cells per se.


[Duodenal duplication: pathologic anatomy and embryogenesis. Case report in an adult].
Duodenal duplication is a rare congenital entity and less than 100 cases have thus far been reported in the literature. This was first described by Sanger in 1880. By definition, they are located in or adjacent to the wall part of the gastrointestinal tract, have smooth muscle in 2 layers and are lined by alimentary tract mucosa. With the case report of a 18 year-old patient with pancreatitis, we expose modern imaging procedure and surgical management. Although the exact etiology of enteric duplications is not known, the two main hypothesis are dysembryogenesis and dysorganogenesis.


The Potential Role of Spa Therapy in Managing Frailty in Rheumatic Patients: A Scoping Review.
Frailty is not limited to the elderly, as patients with rheumatic diseases can also experience this condition. The present scoping review aimed to investigate the possibility of using the health resort setting as an alternative location for managing rheumatic patients with frailty. The research resulted in finding several in vitro, in vivo, and clinical studies, resulting in evidence supporting the effectiveness of spa treatments in reducing pain, improving function, and managing comorbidity in rheumatic diseases. Additionally, spa treatments were demonstrated to modulate the MAPK/ERK pathway and the NF-kB pathway's activation and to reduce proinflammatory molecules' secretion in rheumatic diseases, thus suggesting their potential effective role in the regulation of inflammaging in frailty. Moreover, the health resort setting may offer potential resources to reduce risk factors, such as drug consumption, inactivity, and disease severity, and may serve as a setting for developing prevention protocols for frailty. Future research should explore innovative approaches, such as exercise training and early diagnostics, for the overall management of frailty in rheumatic patients in the spa setting.


BODIPY-conjugated neuropeptide Y ligands: new fluorescent tools to tag Y1, Y2, Y4 and Y5 receptor subtypes.
N-terminal labelled fluorescent BODIPY-NPY peptide analogues were tested in Y1, Y2, Y4 and Y5 receptor-binding assays performed in rat brain membrane preparations and HEK293 cells expressing the rat Y1, Y2, Y4 and Y5 receptors. BODIPY TMR/FL-[Leu31, Pro34]NPY/PYY were able to compete for specific [125][Leu31, Pro34]PYY-binding sites with an affinity similar to that observed for the native peptide at the Y1 (Ki=1-6 nM), Y2 (Ki>1000 nM), Y4 (Ki=10 nM) and Y5 (Ki=1-4 nM) receptor subtypes. BODIPY FL-PYY(3-36) was able to compete for specific Y2 (Ki=10 nM) and Y5 (Ki=30 nM) binding sites, but had almost no affinity in Y1 and Y4 assays. BODIPY FL-hPP was able to compete with high affinity (Ki; 1 and 15 nM) only in Y4 and Y5 receptor-binding assays. BODIPY TMR-[cPP(1-7), NPY(19-23), Ala31, Aib32, Gln34]hPP and BODIPY TMR-[hPP(1-17), Ala31, Aib32]NPY were potent competitors only on specific Y5-binding sites (Ki=0.1-0.6 nM). As expected, these fluorescent peptides inhibited forskolin-induced cAMP accumulation, demonstrating that they retained their agonist properties. When tested in confocal microscopy imaging, fluorescent Y1 and Y5 agonists internalized in a time-dependent manner in Y1 and Y5 transfected cells, respectively. These results demonstrate that BODIPY-conjugated NPY analogues retain their selectivity, affinity and agonist properties for the Y1, Y2, Y4 and Y5 receptor subtypes, respectively. Thus, they represent novel tools to study and visualize NPY receptors in living cells.


Effects of topical travoprost and unoprostone on optic nerve head circulation in normal rabbits.
To evaluate the effect, and the duration of the effect, of topically administrated travoprost and unoprostone on optic nerve head (ONH) circulation in Dutch rabbits. First, travoprost (0.004% solution) or unoprostone (0.12% solution) was unilaterally instilled once, or once daily (travoprost) or twice daily (unoprostone) for 7 days in Dutch rabbits. The ONH tissue blood velocity (NB(ONH)) was measured using the laser speckle method at 30 and 60 min after a single instillation of travoprost or unoprostone, and before and at 1, 6, and 12 hr (travoprost or unoprostone) and 24 hr (travoprost only) after the last instillation of the aforementioned 7-day instillation regimen. Second, similar experiments were conducted with indomethacin (5 mg/kg) pretreatment. Both travoprost and unoprostone significantly increased NB(ONH) only in the treated eyes after a single instillation (p = 0.011 to 0.038); this effect was abolished by indomethacin pretreatment. In the 7-day instillation regimen, NB(ONH) was increased by 11%, 40%, 17%, 16%, and 12% only in the treated eyes just before and at 1, 6, 12, and 24 hr after the final instillation of travoprost, respectively, and increased by 10%, 25%, 13%, and 14% only in the treated eyes just before and at 1, 6, 12 hr after the final instillation of the unoprostone, respectively. Topical travoprost or unoprostone significantly increased the ONH blood velocity with a single instillation and the effect persisted for 24 hr after a 7-day instillation. The effects of these drugs against retinal and ONH circulation are probably associated with the production of endogenous prostaglandins.


Understanding the genesis of psychotic disorder: issues in the prediction and prophylaxis of those at 'ultra-high risk'.
Recent research is starting to identify individuals at an increased risk of developing a psychotic disorder. This review seeks to identify the 'state of the art' with respect to the clinical identification and treatment of individuals at 'ultra-high risk'. The research and clinical literature are reviewed with respect to a wide range of predictors relevant across development. The review draws on evidence from childhood and adolescence to suggest that a range of biological, cognitive, personality, and social features are predictive of, but often not specific to, psychosis within the context of the diathesis-stress model. Much evidence supports the view that environmental stressors act in combination with vulnerability factors to increase risk of transition, often during late adolescence and early adulthood. Recent clinical research has used both retrospective and prospective means of identifying individuals at ultra-high risk and has made substantial gains in predicting transition. The clinical implications of their identification are discussed in the context of current pharmacological and psychosocial treatment studies. However, significant unresolved clinical and ethical issues remain with both types of study.


The PD-1- and LAG-3-targeting bispecific molecule tebotelimab in solid tumors and hematologic cancers: a phase 1 trial.
Tebotelimab, a bispecific PD-1×LAG-3 DART molecule that blocks both PD-1 and LAG-3, was investigated for clinical safety and activity in a phase 1 dose-escalation and cohort-expansion clinical trial in patients with solid tumors or hematologic malignancies and disease progression on previous treatment. Primary endpoints were safety and maximum tolerated dose of tebotelimab when administered as a single agent (n = 269) or in combination with the anti-HER2 antibody margetuximab (n = 84). Secondary endpoints included anti-tumor activity. In patients with advanced cancer treated with tebotelimab monotherapy, 68% (184/269) experienced treatment-related adverse events (TRAEs; 22% were grade ≥3). No maximum tolerated dose was defined; the recommended phase 2 dose (RP2D) was 600 mg once every 2 weeks. There were tumor decreases in 34% (59/172) of response-evaluable patients in the dose-escalation cohorts, with objective responses in multiple solid tumor types, including PD-1-refractory disease, and in LAG-3+ non-Hodgkin lymphomas, including CAR-T refractory disease. To enhance potential anti-tumor responses, we tested margetuximab plus tebotelimab. In patients with HER2+ tumors treated with tebotelimab plus margetuximab, 74% (62/84) had TRAEs (17% were grade ≥3). The RP2D was 600 mg once every 3 weeks. The confirmed objective response rate in these patients was 19% (14/72), including responses in patients typically not responsive to anti-HER2/anti-PD-1 combination therapy. ClinicalTrials.gov identifier: NCT03219268 .


Hemispheric asymmetry in resting motor threshold in major depression.
Previous studies of repetitive transcranial magnetic stimulation suggest a hemispheric imbalance in patients with major depression. These studies report an antidepressant effect by activation of the left prefrontal cortex or inhibition of the right prefrontal cortex. The aim of this study is to investigate interhemispheric differences in cortical excitability in a large sample of patients with major depression. The resting motor threshold (RMT) was measured on 91 patients with treatment-resistant major depression. We controlled for current medication use, gender, age, handedness, and study site. There was no significant difference between the left RMT (55.96 [10.356]) and the right (57.74 [11.359]) (P = 0.131, Wilcoxon matched-pairs test). A multivariate analysis found no significant association between depression scores and right or left RMT. After adjusting for important cofactors, benzodiazepine use was found to be a significant predictor of left RMT (P = 0.017, linear regression) and right RMT (P = 0.007, linear regression). Our results do not support the existence of an interhemispheric imbalance of cortical excitability in depressed patients. Benzodiazepine use was found to raise both the left and right RMT.


Integrated proteomic and metabolomic analyses reveal significant changes in chloroplasts and mitochondria of pepper (Capsicum annuum L.) during Sclerotium rolfsii infection.
Infection by Sclerotium rolfsii will cause serious disease and lead to significant economic losses in chili pepper. In this study, the response of pepper during S. rolfsii infection was explored by electron microscopy, physiological determination and integrated proteome and metabolome analyses. Our results showed that the stomata of pepper stems were important portals for S. rolfsii infection. The plant cell morphology was significantly changed at the time of the fungal hyphae just contacting (T1) or surrounding (T2) the pepper. The chlorophyll, carotenoid, and MDA contents and the activities of POD, SOD, and CAT were markedly upregulated at T1 and T2. Approximately 4129 proteins and 823 metabolites were clearly identified in proteome and metabolome analyses, respectively. A change in 396 proteins and 54 metabolites in pepper stem tissues was observed at T1 compared with 438 proteins and 53 metabolites at T2. The proteins and metabolites related to photosynthesis and antioxidant systems in chloroplasts and mitochondria were disproportionally affected by S. rolfsii infection, impacting carbohydrate and amino acid metabolism. This study provided new insights into the response mechanism in pepper stems during S. rolfsii infection, which can guide future work on fungal disease resistance breeding in pepper.


New advances in insulin treatment of diabetes: overcoming barriers.
To discuss the optimal role of insulin in the treatment of patients with diabetes mellitus. We review the complications of diabetes, highlight the attempts to improve control of plasma glucose, and summarize current recommendations for use of insulin in clinical practice. With the strict new guidelines for the diagnosis of diabetes issued by the American Diabetes Association in July 1997--a plasma glucose level of 126 mg/dL (7 mmol/L) rather than 140 mg/dL (7.8 mmol/L)--an additional 8 million persons will be diagnosed this year, and diabetes and its complications will be at the forefront of public health concerns. Strong evidence indicates that with reduction of plasma glucose levels and tight control of glycohemoglobin, the rate of complications can be considerably decreased. Although insulin replacement therapy is well known to be both the best and the most cost-effective way to control glucose levels in patients with type 1 diabetes, studies have no shown that those with type 2 diabetes can likewise benefit from appropriate insulin therapy. Other investigations have indicated that coronary events are as likely to occur in patients with recently discovered impaired glucose tolerance as in patients with known diabetes (at a 2-hour postprandial glucose threshold of 96 mg/dL [5.3 mmol/L]). Such finding suggest that there may be no such thing as "borderline diabetes" and give impetus to the search for improved types of insulin to treat all patients with diabetes. A recent candidate is the new insulin analogue lispro, which was developed by recombinant DNA techniques, with its design influenced by the structural analogy to another endogenous hormone, insulin-like growth factor I. Lispro is a fast-acting, rapidly dissipating insulin formulation. This profile allows lispro to be given less than 15 minutes before a meal, yet with little risk of postprandial hypoglycemia because its high mealtime peak is followed by rapid disappearance from the bloodstream. Lispro has been shown to improve postprandial control of plasma glucose and to decrease the occurrence of hypoglycemia episodes in comparison with regular insulin. Because of its stability and pharmacokinetic properties, lispro can also be used in insulin pump therapy.


MFS transporter from Botrytis cinerea provides tolerance to glucosinolate-breakdown products and is required for pathogenicity.
Glucosinolates accumulate mainly in cruciferous plants and their hydrolysis-derived products play important roles in plant resistance against pathogens. The pathogen Botrytis cinerea has variable sensitivity to glucosinolates, but the mechanisms by which it responds to them are mostly unknown. Exposure of B. cinerea to glucosinolate-breakdown products induces expression of the Major Facilitator Superfamily transporter, mfsG, which functions in fungitoxic compound efflux. Inoculation of B. cinerea on wild-type Arabidopsis thaliana plants induces mfsG expression to higher levels than on glucosinolate-deficient A. thaliana mutants. A B. cinerea strain lacking functional mfsG transporter is deficient in efflux ability. It accumulates more isothiocyanates (ITCs) and is therefore more sensitive to this compound in vitro; it is also less virulent to glucosinolates-containing plants. Moreover, mfsG mediates ITC efflux in Saccharomyces cerevisiae cells, thereby conferring tolerance to ITCs in the yeast. These findings suggest that mfsG transporter is a virulence factor that increases tolerance to glucosinolates.


The fine structure of the ventral intersegmental abdominal muscles of the insect Rhodnius prolixus during the molting cycle. II. Muscle changes in preparation for molting.
The development of the ventral intersegmental abdominal muscles of Rhodnius prolixus is triggered by feeding. The early muscle (1 day after feeding) contains essentially nonstriated fibrils. However, in cross-sections, areas indicating early I bands, Z lines, and A bands can be recognized. Interdigitating thick and thin myofilaments do not assemble into a precise lattice until sometime between 4 and 5 days after feeding. As development continues, the number of fibrils increases, the region corresponding to the Z line increases in density, and the fibrils contain more recognizable striations. The newly formed fibrils broaden as myofilaments are added peripherally. At all stages throughout development, the ratio of thin to thick myofilaments is always 6:1. The formation of fibrils in the abdominal muscles of Rhodnius is different from that in chick embryo skeletal muscle. The major differences are that at all stages in Rhodnius there are (1) a constant ratio of thin to thick myofilaments, and (2) detectable Z-line material. Other findings in Rhodnius suggest (1) that fusion of mononucleated cells with the multinucleated muscle cell occurs, (2) that microtubules develop in the tendon cell concomitantly with development of myofibrils in the associated muscle cell, and (3) that filaments 55A in diameter aggregate into microtubules.


Histologic evaluation of periodontal implants in a biologically "closed" model.
To study the biologic response, three commercial calcium phosphate implant materials (Calcitite, Periograf, Synthograft) were implanted in cuspid root "windows" in four beagle dogs. No implant material was placed in the fourth cuspid window which served as a control. Following mucoperiosteal flap elevation, windows were chiseled in bone to the root surface which allowed implant particles to contact bone, fibrous connective tissue, cementum and dentin without exposure to the oral environment. Animals were sacrificed after 1, 3, 5 and 6 months. Histologically, all implant materials were well tolerated. At 1 month, implant particles were surrounded by fibrous tissue. Fibrous tissues filled the control defect. At 3 months, implant sites exhibited partial bony repair with connective tissue surrounding implant particles. A ring of osteoid surrounded Synthograft particles. Control defect repair was complete. At 5 and 6 months, implant sites exhibited advanced, though incomplete, bony repair. New bone encompassed the Synthograft particles. It was concluded: Control sites healed most rapidly. Calcitite and Periograft were well tolerated space occupiers. Synthograft was a nidus for bone deposition.


Combating liver cancer through GO-targeted biomaterials.
Hydroxycamptothecin (HCPT) is a topoisomerase I inhibitor, and it has been widely used clinically in the treatment of primary liver cancer, gastric cancer, and other tumors. The clinical application of HCPT is limited by its water solubility, and it has certain toxicity to patients with tumor. Therefore, the effective tumor site accumulation of HCPT is necessary. This work studied the inhibitory effect of HCPT on the proliferation and migration of human liver cancer cells (HepG-2) and used carboxymethyl chitosan (CMC) and hyaluronic acid (HA) to modify graphene oxide (GO) as nano-carrier materials, which load HCPT to achieve a drug delivery system for liver tumors with good biocompatibility and high drug loading. HCPT can significantly inhibit proliferation and migration of HepG-2, enhance the release of reactive oxygen species, reduce mitochondrial membrane potential, and induce apoptosis. The GO-CMC-HA/HCPT drug delivery system enabled HepG-2 to uptake more HCPT, thereby inhibiting its proliferation and improving the efficacy of HCPTin vivoandin vitro. This study explored a potential therapy strategy by preparing a GO-based tumor-targeted drug delivery system.


Effect of expansion of extracellular fluid volume on renal phosphate handling.
To examine the specific effect of extracellular fluid (ECF) volume expansion on phosphate excretion studies were performed in thyroparathyroidectomized dogs receiving saline solution intravenously. The natriuresis resulting from ECF volume expansion was consistently accompanied by an increase in phosphate excretion. The possible role of increased filtered load of phosphate was eliminated in experiments in which the filtered load of phosphate was reduced by acute reduction in the glomerular filtration rate. Despite considerable reductions in filtered phosphate, ECF volume expansion resulted in a consistent increase in phosphate excretion. Furthermore, the possible contribution of alteration in blood composition was investigated in experiments in which saline was infused during thoracic inferior vena cava constriction. In these experiments saline infusion failed to increase sodium or phosphate excretion. Cessation of saline infusion and release of caval constriction resulted in a prompt natriuresis and increased phosphate excretion. It is concluded from these studies that extracellular fluid volume expansion results in an increased phosphate excretion in the parathyroidectomized dog. This effect is the specific consequence of ECF volume expansion and is not due to increase in the filtered load of phosphate or alterations in blood composition.