Effects of platelet-derived growth factor on 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in cultured human fibroblasts.
Studies were conducted to delineate in detail the effects of platelet-derived growth factor (PDGF) upon 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity in cultured human fibroblasts. PDGF has a variety of coordinated effects on low density lipoprotein (LDL) and cholesterol metabolism and on DNA synthesis in these cells. We have previously shown that there is a critical time period, 10 to 20 hours after PDGF addition to quiescent cells, when mevalonate (MVA) is required for DNA synthesis to occur. The present studies demonstrate that PDGF produces a biphasic stimulation of reductase activity in cultured fibroblasts: a peak at 4 to 6 hours, followed by a decline, and then a second smaller peak at 24 hours, concurrent with DNA synthesis. The stimulation of both peaks was PDGF concentration-dependent, although quantitative differences were observed. Proportionally similar, LDL concentration-dependent reductions in both peaks of reductase activity were also seen. PDGF stimulated cholesterol synthesis from acetate in intact cells reaching peak values after 24 to 28 hours. The two peaks of reductase activity stimulated by PDGF neither coincide with the critical time period when mevalonate is needed for later DNA synthesis to occur, nor reflect the pattern of cholesterol synthesis within the cells.


Safety evaluation of the food enzyme with 4-α-d-{(1->4)-α-d-glucano}trehalose trehalohydrolase and (1->4)-α-d-glucan 1-α-d-glucosylmutase activities from the Gryllotalpicola ginsengisoli strain S34.
The food enzyme with 4-α-d-{(1->4)-α-d-glucano}trehalose trehalohydrolase ((4-α-d-[(1,4-α-d-glucano]trehalose glucanohydrolase (trehalose-producing)), EC 220.127.116.11) and (1->4)-α-d-glucan 1-α-d-glucosylmutase ((1,4)-α-d-glucan 1-α-d-glucosylmutase, EC 184.108.40.206) activities is produced with a non-genetically modified microorganism Gryllotalpicola ginsengisoli (formally Arthrobacter ramosus) by Hayashibara Co., Ltd. The applicant states that the food enzyme will not be placed on the open market, but will only be used in-house in starch processing for trehalose production. Since residual amounts of total organic solids are removed by the purification steps applied during the production of trehalose, toxicological studies were considered not necessary and no dietary exposure was calculated. Similarities of the amino acid sequences to those of known allergens were searched and no matches were found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood to occur is considered to be low. Based on the data provided, the Panel concluded that the food enzyme does not give rise to safety concerns under the intended conditions of use.


Serological reactivity to Borrelia burgdorferi sensu lato in dogs and horses from distinct areas in Romania.
Lyme disease is a perfect model of the complex relationship between host, vector, and the vector-borne bacteria. Both dogs and horses in Romania are exposed to infection. The aim of the present study was to assess the seroreactivity against Borrelia burgdorferi sensu lato in dogs and horses from different regions of Romania. 276 samples from dogs and 260 samples from horses located in different regions of Romania were analyzed by ELISA and IFA, respectively. The effect of several factors potentially affecting seroreactivity (location, age, gender, occupation, and vector exposition risk) was evaluated using Fisher's exact test (R 2.12.0). The overall prevalence of anti-Borrelia antibodies was 6.52% (18/276) in dogs, with a significantly higher positivity (46.15%, 6/13, p = 0.0005) recorded in a midcountry region. Seroreactivity was correlated with occupation, with working dogs being more exposed. The results may indicate that Lyme borreliosis foci are restricted to small areas, but further studies on Borrelia prevalence in tick populations are needed to confirm this hypothesis. In horses, a global seroprevalence of 11.92% (31/260) was observed. No correlations were found between positive results and age, sex, county, or occupation. This is the first serological survey on antibodies to B. burgdorferi sensu lato in Romanian dogs and horses.


Alleviation of the Byproducts Formation Enables Highly Efficient Biosynthesis of Rosmarinic Acid in Saccharomyces cerevisiae.
Rosmarinic acid as a polyphenolic compound has great values in the pharmaceutical, cosmetic, and food industries. To achieve efficient biosynthesis of rosmarinic acid, the major obstacles such as imbalanced metabolic flux among branching pathways and substrate promiscuity of pathway enzymes should be eliminated. Here, a rosmarinic acid producing Saccharomyces cerevisiae strain was constructed by introducing codon optimized d-lactate dehydrogenase gene mutant (OD-LDHY52A), 4-coumarate CoA ligase gene (OPc4CL2), and rosmarinic acid synthase gene (OMoRAS) into a previously constructed caffeic acid hyper-producer. To identify the metabolic bottleneck, the substrate specificity of OPc4CL2 and OMoRAS was figured out by bioconversion experiments and HPLC-MS/MS analysis. Subsequently, the byproducts formation was alleviated by removing prephenate dehydratase and tuning down the expression level of OPc4CL2. The final strain YRA113-15B produced 208 mg/L rosmarinic acid in a shake-flask culture (a 63-fold improvement over the initial strain), which was the highest rosmarinic acid titer by engineered microbial cells reported to date. This work provides a promising platform for fermentative production of rosmarinic acid and offers a strategy to overcome the intrapathway competition.


[Extra-lemniscal afferent projections of dorsal spinal cord nuclei to the ventrobasal nuclear complex structure in the contralateral optic thalamus].
It has been found that section of half the midbrain tegmentum in cats failed to prevent the afferent somatosensory projections from the foreleg to the ventrobasal nuclear complex of the contralateral thalamus. Specific evoked responses to the stimulation of the contralateral foreleg were recorded in this structure. These specific EP have the same latency as "lemniscal responses" (4-5 ms) and diminish the amplitude and duration of both components of the responses. Simultaneously, we have observed terminal axonal degeneration into the ventrobasal nuclear complex of the thalamus 5-7 days after the section of the contralateral midbrain tegmentum, using the electron microscopy method. All the results obtained indicate that the dorsal column nuclei have extra-lemniscal afferent connections with ventrolateral nuclear complex of the contralateral thalamus. These connections ascend in the back parts of the brainstem ipsilaterally to the corresponding pair of the dorsal column nuclei and rostrally to the midbrain on the contralateral side.


Diagnostic efficiency and quality indexes of several markers of renal function for detecting the loss of parenchyma in paediatric patients.
We analysed a large sample of children diagnosed with urinary tract malformations and/or infections and calculated diagnostic efficiency and quality indexes for five different functional markers, with the goal of testing which is the most sensitive for detecting a loss of renal parenchyma. Ours was a cross-sectional retrospective study in which the clinical histories of 179 paediatric patients (91 male and 88 female) were evaluated. In 102 of these patients (57%), a scintigraphy revealed loss of parenchyma. The most commonly observed morphological type of damage was renal scarring. All patients had undergone at least one desmopressin urine concentration test. We also analysed albumin/creatinine and N-acetyl-glucosaminidase (NAG)/creatinine ratios, glomerular filtration rate (GFR), and urine volume. By distributing patients according to normal/abnormal scintigraphy, we observed statistically significant differences between the two groups in maximum urine osmolality and GFR. Urine volume was elevated in 31.3% of cases (sensitivity: 37.9%; specificity: 81.8%) and 24% had a defect in renal concentrating ability (sensitivity: 30.4%; specificity: 84.8%). Urinary albumin excretion was high in 12.2% of patients, and 7.2% had a high NAG/creatinine ratio. GFR was low in only 5.7% of patients. These last two markers were the least sensitive but most specific for detecting a loss of renal parenchyma (100%). In our study, the most sensitive functional tests for detecting the loss of renal parenchyma were the two that take into account the ability of the kidney to manage water, i.e. urine volume and maximum urine osmolality. These two tests had specificity >80%. However, the maximum specificity was obtained by the NAG/creatinine ratio and GFR, which were, conversely, the least sensitive tests. A normal GFR does not necessarily show normal renal function.


Antituberculosis thiophenes define a requirement for Pks13 in mycolic acid biosynthesis.
We report a new class of thiophene (TP) compounds that kill Mycobacterium tuberculosis by the previously uncharacterized mechanism of Pks13 inhibition. An F79S mutation near the catalytic Ser55 site in Pks13 conferred TP resistance in M. tuberculosis. Overexpression of wild-type Pks13 resulted in TP resistance, and overexpression of the Pks13(F79S) mutant conferred high resistance. In vitro, TP inhibited fatty acyl-AMP loading onto Pks13. TP inhibited mycolic acid biosynthesis in wild-type M. tuberculosis, but it did so to a much lesser extent in TP-resistant M. tuberculosis. TP treatment was bactericidal and equivalent to treatment with the first-line drug isoniazid, but it was less likely to permit emergent resistance. Combined isoniazid and TP treatment resulted in sterilizing activity. Computational docking identified a possible TP-binding groove within the Pks13 acyl carrier protein domain. This study confirms that M. tuberculosis Pks13 is required for mycolic acid biosynthesis, validates it as a druggable target and demonstrates the therapeutic potential of simultaneously inhibiting multiple targets in the same biosynthetic pathway.


Detection and measurement of multivalent antigen in the presence of low affinity antigen-antibody complexes.
Multivalent antigen which is bound to antibody can be difficult to measure and detect. There are 2 reasons for this phenomenon: (a) The antibody can mask the antigen so that it cannot react with antibody in the in vitro test system and (b) Complexes of multivalent antigen and antibody can precipitate spontaneously. In this study solutions to these problems were developed for the special case in which the antigen-antibody bond is of low affinity. The masking effect of low affinity antibody could be overcome by using a very high affinity antibody (K = 10(10) M-1) in the test system. The precipitation problem could be minimized by using fresh or thawed fresh frozen serum. In cases where antigen detection, rather than quantitation, is required, analysis of precipitates may be preferable because antigen is often selectively concentrated in the redissolved precipitates over serum values.


Imprint of monsoonal patterns on the fish assemblage in coastal waters of south-east India: a case study.
The composition, species richness and diversity of a coastal fish assemblage from the Kalpakkam coast of south-east India are described along with temporal distribution patterns related to seasonal fluctuations in dissolved oxygen, salinity, pH, chlorophyll-a, phytoplankton and zooplankton species richness and density. A total of 244 fish species belonging to 21 orders, 87 families and 163 genera were recorded. The fish assemblage was dominated by reef-associated species, followed by demersal species. The majority of the species (63%) are widely distributed in the western Indo-Pacific as well as in the central Indo-Pacific. Jaccard's coefficient analysis showed three distinct seasonal patterns of fish occurrence: pre-monsoon (PrM), monsoon (M) and post-monsoon (PoM). The maximum number of species was during the PrM period, followed by the PoM and M periods. Species occurrence analysis showed Sardinella longiceps to be dominant during PrM and M periods, Leiognathus dussumieri during the M period and Secutor insidiator and Secutor ruconius during the M and PoM periods. Canonical correspondence analysis indicated that salinity and rainfall were the two most influential environmental factors strongly correlated with temporal variation in the fish assemblage. The physico-chemical conditions, in combination with factors such as greater food availability and shelter, might control the seasonal local distribution of the ichthyofauna in these Indian coastal waters.


Three-Dimensional Bioprinting and Its Potential in the Field of Articular Cartilage Regeneration.
Three-dimensional (3D) bioprinting techniques can be used for the fabrication of personalized, regenerative constructs for tissue repair. The current article provides insight into the potential and opportunities of 3D bioprinting for the fabrication of cartilage regenerative constructs. Although 3D printing is already used in the orthopedic clinic, the shift toward 3D bioprinting has not yet occurred. We believe that this shift will provide an important step forward in the field of cartilage regeneration. Three-dimensional bioprinting techniques allow incorporation of cells and biological cues during the manufacturing process, to generate biologically active implants. The outer shape of the construct can be personalized based on clinical images of the patient's defect. Additionally, by printing with multiple bio-inks, osteochondral or zonally organized constructs can be generated. Relevant mechanical properties can be obtained by hybrid printing with thermoplastic polymers and hydrogels, as well as by the incorporation of electrospun meshes in hydrogels. Finally, bioprinting techniques contribute to the automation of the implant production process, reducing the infection risk. To prompt the shift from nonliving implants toward living 3D bioprinted cartilage constructs in the clinic, some challenges need to be addressed. The bio-inks and required cartilage construct architecture need to be further optimized. The bio-ink and printing process need to meet the sterility requirements for implantation. Finally, standards are essential to ensure a reproducible quality of the 3D printed constructs. Once these challenges are addressed, 3D bioprinted living articular cartilage implants may find their way into daily clinical practice.


A liquid crystal atmospheric turbulence simulator.
In the paper, a method to calculate the time evolution turbulence wavefronts based on the covariance method is theoretically presented in detail. According to it, the time-evolution wavefronts disturbed by atmospheric turbulence were experimentally generated by our LC atmospheric turbulence simulator (ATS) based on liquid crystal on silicon (LCOS) with high pixel density, and measured with a wavefront sensor. The advantage of such a LC ATS over a conventional one is that it is flexible with considering the weather parameters of wind speed and C(n) (2), and is relatively easy to control.


Triterpenes as potential dimerization inhibitors of HIV-1 protease.
HIV-1 protease is a homodimeric enzyme. A beta-sheet consisting of the four terminal segments provides the main driving force for dimerization of the per se inactive protomers. Several short peptides with sequences related to the terminal sequences of the protease are able to inhibit dimerization by blocking the 'interface' part of the monomers. From the structures of such inhibitory peptides a 'pharmacophore' could be derived. By using a prominent distance from this pharmacophore scaffold for a library search (Cambridge Structural Database), non-peptide inhibitors of HIV-1 protease with polycyclic triterpene structure could be found. The IC50 constants of these compounds are near 1 microM. One of the triterpenes, the ursolic acid (Ki = 3.4 microM), was further kinetically analysed (according to Zhang). The shape of the graph confirms the expected mechanism of dimerization inhibition.


Next Generation of the Java Image Science Toolkit (JIST): Visualization and Validation.
Modern medical imaging analyses often involve the concatenation of multiple steps, and neuroimaging analysis is no exception. The Java Image Science Toolkit (JIST) has provided a framework for both end users and engineers to synthesize processing modules into tailored, automatic multi-step processing pipelines ("layouts") and rapid prototyping of module development. Since its release, JIST has facilitated substantial neuroimaging research and fulfilled much of its intended goal. However, key weaknesses must be addressed for JIST to more fully realize its potential and become accessible to an even broader community base. Herein, we identify three core challenges facing traditional JIST (JIST-I) and present the solutions in the next generation JIST (JIST-II). First, in response to community demand, we have introduced seamless data visualization; users can now click 'show this data' through the program interfaces and avoid the need to locating files on the disk. Second, as JIST is an open-source community effort by-design; any developer may add modules to the distribution and extend existing functionality for release. However, the large number of developers and different use cases introduced instability into the overal JIST-I framework, causing users to freeze on different, incompatible versions of JIST-I, and the JIST community began to fracture. JIST-II addresses the problem of compilation instability by performing continuous integration checks nightly to ensure community implemented changes do not negatively impact overall JIST-II functionality. Third, JIST-II allows developers and users to ensure that functionality is preserved by running functionality checks nightly using the continuous integration framework. With JIST-II, users can submit layout test cases and quality control criteria through a new GUI. These test cases capture all runtime parameters and help to ensure that the module produces results within tolerance, despite changes in the underlying architecture. These three "next generation" improvements increase the fidelity of the JIST framework and enhance utility by allowing researchers to more seamlessly and robustly build, manage, and understand medical image analysis processing pipelines.


Poly(A) polymerase and poly(g) polymerase in wheat chloroplasts.
Extracts of wheat chloroplasts contain a poly(A) polymerase which can polymerize AMP residues from ATP onto an RNA primer. Whole extracts of wheat leaves also contain another poly(A) polymerase which is present in much larger amount and is probably derived from the nuclei. Both polymerases can utilize as primer poly(A), poly(C), transfer RNA, and ribosomal RNA, but only the chloroplast polymerase can utilize poly(U) and poly(G). Both enzymes have a specific requirement for ATP. Extracts of wheat chloroplasts contain, in addition to the poly(A) polymerase, a poly(G) polymerase which can polymerize GMP residues from GTP onto primers such as poly(G), poly(A), or ribosomal RNA. The poly(G) polymerase cannot utilize ATP but can slowly polymerize CMP from CTP. When the two chloroplast polymerases are present together in an in vitro incubation with ATP plus GTP and poly(A), the polymerization product is a mixed poly(A,G) tract.


Effect of nicotine on gastric mucosal blood flow and acid secretion.
The effects of an intravenous infusion of nicotine at a dose of 2 . 5, 5 . 0, 7 . 5, or 10 . 0 micrograms kg-1h-1 on pentagastrin-stimulated gastric mucosal blood flow and acid secretion were investigated in eight healthy male non-smokers. Each dose was tested in two males. Gastric neutral red clearance served as a measure of mucosal blood flow. Nicotine reduced volume secretion, acid secretion, and neutral red clearance in a dose dependent manner. In five healthy male smokers smoking of five cigarettes per two hours induced similar changes to the intravenous infusion of 5 micrograms kg-1h-1 nicotine. As volume secretion was inhibited more than neutral red clearance, it is concluded that nicotine increases blood supply to the gastric mucosa relatively to the reduced gastric secretion. Nicotine is either not associated with the development of peptic ulcers, or it exerts its ulcerogenic action via other mechanisms than change of acid secretion and gastric mucosal blood flow.


Ion-shedding zinc oxide nanoparticles induce microglial BV2 cell proliferation via the ERK and Akt signaling pathways.
Given the wide application of zinc oxide nanoparticles (ZnO NPs), the health hazards of these particles have attracted extensive worldwide attention. Many more studies on the biological interactions of ZnO NPs have been performed in recent years. In this study, we focused on the biological effects on BV2 microglial cells induced by ZnO NPs at non- or sub-toxic concentrations. We found that ZnO NPs at a concentration of 5 μg/ml could significantly activate cell proliferation, while ZnO NPs at other concentrations did not. We also found that ZnO NPs induced microglial cell activation in a time-dependent manner. Moreover, a potent increase in the ratio of cells in S phase at all ZnO NPs concentrations was observed in a cell cycle analysis. Using inductively coupled plasma mass spectrometry (ICP-MS) and immunocytochemistry techniques, we demonstrated that ZnO dissolution could occur in the culture medium and in the lysosomes of BV2 cells. ZnO NPs significantly induced the phosphorylation of ERK and Akt, which might be involved in promoting cell proliferation. In conclusion, our results demonstrated that ZnO NPs induced BV2 microglial cell proliferation probably via the release of zinc ions from ZnO, which could then activate the ERK and Akt signaling pathways.


Absence of linkage between chromosome 11p15 markers and manic-depressive illness in a Belgian pedigree.
The original finding of genetic linkage in an Old Order Amish pedigree has been contradicted by the results of several subsequent studies. Using the same genetic parameter values, diagnostic criteria, and 11p15 genetic markers as those used to study the initial Amish population, the authors performed a linkage study of a four-generation informative pedigree in Belgium. Recombinant DNA technology was used to analyze three markers for the chromosome 11p15 location: the genes for tyrosine hydroxylase (TH) and insulin (INS) and the c-Harvey-ras oncogene (HRAS). Diagnoses of the relatives of a proband with bipolar affective disorder were determined with the Schedule for Affective Disorders and Schizophrenia--Lifetime Version and based on the Research Diagnostic Criteria. Relatives were considered affected if they had bipolar disorder, unipolar disorder, or cyclothymia; a diagnostic hierarchy was developed to include unipolar disorder and cyclothymia in the linkage analysis. Pairwise analyses of the disease locus and each of the three polymorphisms excluded the possibility of close linkage between manic-depressive illness and the three chromosome 11p15 markers. Multipoint linkage analysis combining the information from all three genes also excluded linkage. The conflict between the original results from the Amish study and the many negative reports on chromosome 11 linkage of manic-depression has been interpreted as indicating genetic heterogeneity, but heterogeneity has not been documented for the 11p15 locus. Conversely, the linkage approach has major drawbacks, so other genetic strategies should also be considered.


Audit from preschool developmental surveillance of vision, hearing, and language referrals.
Referrals from preschool medical examinations were followed up for two years to assess attendance rate, waiting time for appointment, appropriateness of the referral, the diagnosis and management of the condition. Altogether 184 children were referrals for ophthalmology, 285 for audiology, and 195 for speech therapy. The median waiting time for an appointment was 46 days in ophthalmology, 175 days in audiology, and 83 days in speech therapy. The poorest attendance rate was identified in speech therapy (75%). Approximately 60% of examined children had a justified referral to ophthalmology and 20% had a clear defect. Over half the children in audiology (55%) had an altered impedance or hearing impairment. Of those with a hearing problem kept under review only half improved spontaneously. In speech therapy 80% of those assessed had a language problem. Many health problems were detected for which parents were unaware or did not use the service. Parental awareness alone will not uncover the sizable level of lingual and sensorial problems in inner city areas. This audit identified specific deficiencies in the provision of services and a number of organisational changes are suggested to improve their effectiveness.


[The use of reflexotherapy in the initial forms of cerebrovascular insufficiency in persons who suffered as a consequence of the accident at the Chernobyl Atomic Electric Power Station].
A comprehensive evaluation has been done in 120 patients presenting with primary forms of cerebrovascular insufficiency (PFCVI), who had become victims of Chernobyl accident. PFCVI diagnosis was made on the basis of findings from the clinical investigation, rheoencephalography, electroencephalography, echoencephalography, iridodiagnosis, electropuncture diagnosis as recommended by Nakatani, variation pulsometry, study into the vegetative nervous system. Functional inadequacies were disclosed in the bodily regulatory-adaptive system, specifically, the system of regulation of cerebral circulation. Cerebral symptoms in PFCVI are a reflection, first of all, of the brain nonspecific systems dysfunction. All this was taken into account in working out schemes of treatment for the above-named category of patients, incorporating lasero- and acupuncture. Exposure to the treatments described resulted in a noticeable improvement in 78% of the patients, and improvement in the rest of the series. High efficiency of the proposed approach to the treatment of PFCVI patients having become victims of Chernobyl accident suggests its pathogenetic significance.


Advances in Skin Tissue Engineering and Regenerative Medicine.
There are an estimated 500,000 patients treated with full-thickness wounds in the United States every year. Fire-related burn injuries are among the most common and devastating types of wounds that require advanced clinical treatment. Autologous split-thickness skin grafting is the clinical gold standard for the treatment of large burn wounds. However, skin grafting has several limitations, particularly in large burn wounds, where there may be a limited area of non-wounded skin to use for grafting. Non-cellular dermal substitutes have been developed but have their own challenges; they are expensive to produce, may require immunosuppression depending on design and allogenic cell inclusion. There is a need for more advanced treatments for devastating burns and wounds. This manuscript provides a brief overview of some recent advances in wound care, including the use of advanced biomaterials, cell-based therapies for wound healing, biological skin substitutes, biological scaffolds, spray on skin and skin bioprinting. Finally, we provide insight into the future of wound care and technological areas that need to be addressed to support the development and incorporation of these technologies.


Temporal Trends and Hospital Variation in Mineralocorticoid Receptor Antagonist Use in Veterans Discharged With Heart Failure.
Despite concerns about mineralocorticoid receptor antagonist therapies (MRAs) underuse and misuse in patients with heart failure, temporal and institutional variations of MRA prescription have not been reported. We studied a national sample of veterans hospitalized for heart failure between 2003 and 2009 and left ventricular ejection fraction <40%. We identified ideal and non-ideal candidates for MRA therapy based on American College of Cardiology/American Heart Association guidelines. We measured temporal trends and hospital variation of MRA prescriptions within 90 days after discharge. We determined the median odds ratio (MOR), a measure of the relative odds of an MRA prescription for 2 individuals with similar characteristics discharged at 2 randomly selected hospitals. From 37 126 patients (n=131 hospitals), 9355 were ideal-MRA candidates, and 4056 were non-ideal candidates. Among ideal candidates, 36% received an MRA, but there was a decline in use (41% in 2003 to 31% in 2009, P<0.001). Of non-ideal candidates, 27% received an MRA with a decline in use (34% in 2003 to 22% in 2009, P<0.001). Hospital MRA prescription ranged from 0% to 71% for ideal candidates and 0% to 100% for non-ideal candidates. The median odds ratios of MRA prescription for ideal and non-ideal candidates were 1.44 and 1.36, respectively; a median odds ratio >1.2 indicates significant practice-level variation. There was decreasing MRA use between 2003 and 2009 with wide institutional variation in MRA prescription, which suggests opportunities for improvement to stimulate MRA use in ideal candidates while further reducing use in those with contraindications.


Characterization of expression of glycan ligands for Siglec-F in normal mouse lungs.
Sialic acid-binding immunoglobulin-like lectin (Siglec)-F, an inhibitory receptor on mouse eosinophils, preferentially recognizes the glycan ligand 6'-sulfated sialyl Lewis X, but little is known about the requirements for its lung expression. RT-PCR and immunohistochemistry were used to detect and localize the sulfotransferase keratin sulfate galactose 6-O sulfotransferase (KSGal6ST, also known as carbohydrate sulfotransferase 1; gene name, Chst1) that is putatively required for 6'-sulfated Sialyl Lewis X synthesis. RT-PCR detected the greatest constitutive expression of Chst1 in lung, liver, and spleen tissue. Immunohistochemistry localized the expression of KSGal6ST in lung tissue primarily to airway epithelium. Siglec-F-Ig fusion protein selectively bound in a similar pattern, and was unaffected in lung tissue treated with methanol or deficient in Type 2 α2,3 sialyltransferase (St3gal2), but was eliminated by proteinase K or sialidase, and was absent in tissue deficient in the Type 3 α2,3 sialyltransferase (St3gal3). Binding of the Siglec-F-Ig fusion protein was similar in pattern to, and completely blocked by, a plant lectin recognizing α2,3-linked sialic acid. Thus, α2,3-linked sialic acid-containing glycoprotein Siglec-F ligands and the enzymes required for their synthesis are constitutively expressed in murine lungs, especially by airway epithelium. St3gal3, but not St3gal2, is required for constitutive Siglec-F ligand synthesis. The survival of eosinophils entering the lung may be shortened by encountering these Siglec-F sialoside ligands.


Lambs fed protein or energy imbalanced diets forage in locations and on foods that rectify imbalances.
Ruminants eat a variety of foods from different locations in the environment. While water, cover, social interactions, and predators are all likely to influence choice of foraging location, differences in macronutrient content among forages may also cause ruminants to forage in different locations even during a meal. We hypothesized that lambs forage at locations containing foods that complement their basal diet and meet their nutritional needs. Based on this hypothesis, we predicted that lambs (n=12) fed a basal diet low in protein and high in energy would forage where a high-protein food (Food P) was located, and that lambs (n=12) fed a basal diet low in energy and high in protein would forage where a high-energy food (Food E) was located. Food P was a ground mixture of blood meal (50%), grape pomace (30%), and alfalfa (20%) that contained 47% crude protein (CP) and 2.211 Mcal/kg digestible energy (DE). Food E was a ground mixture of cornstarch (50%), grape pomace (30%), and rolled barley (20%) that contained 6% CP and 3.07 Mcal/kg DE. Food P provided 212 g CP/Mcal DE, whereas Food E provided 20 g CP/Mcal DE. Lambs growing at a moderate rate require 179 g CP and 3.95 Mcal DE. During Trial 1, we determined if lambs foraged to correct a nutrient imbalance, and if they preferred a variety of foods (Foods P and E) to only one food at a location (Food P or E). During Trial 2, we determined if nutrient-imbalanced lambs foraged in the location with the food that corrected the imbalance when the location of the foods changed daily. During Trial 3, lambs were offered familiar foods (Foods P and E) at the location furthest - and novel foods (wheat and soybean meal) at the location nearest - the shelter of their pen. During all three trials, lambs foraged most at the location with the food that contained the highest concentration of the macronutrient lacking in their basal diet, but they always ate some of both foods. Lambs did not feed exclusively at the location with a variety of foods (P and E). Rather, they fed at the location nearest the shelter that contained the macronutrient lacking in their diet. As availability of the food with the needed macronutrient declined in one location, lambs moved to the nearest location that had food with the needed macronutrient. When food that complemented their basal diet was moved to a different location, lambs foraged in the new location. Collectively, these results show that lambs challenged by imbalances in energy or protein selected foods and foraging locations that complemented the nutrient content of their macronutrient imbalanced basal diets.


Pre-dopa Deep Brain Stimulation: Is Early Deep Brain Stimulation Able to Modify the Natural Course of Parkinson's Disease?
Deep brain stimulation (DBS) is an established therapy for the management of Parkinson's disease (PD). However, DBS is indicated as the disease progresses and motor complications derived from pharmacological therapy arise. Here, we evaluate the potential of DBS prior to levodopa (L-Dopa) in improving quality of life (QoL), challenging the state of the art for DBS therapy. We present data on clinical manifestation, decision finding during early indication to DBS, and trajectories after DBS. We further discuss current paradigms for DBS and hypothesize on possible mechanisms. Six patients, between 50 and 67 years old, presenting at least 5 years of PD symptoms, and without L-Dopa therapy initiation, received subthalamic nucleus (STN) DBS implantation. In the six PD cases, indication for DBS was not driven by motor complications, as supported by current guidelines, but by relevant QoL impairment and patient's reluctance to initiate L-Dopa treatment. All patients treated with STN-DBS prior to L-Dopa presented improvement in motor and non-motor symptoms and significant QoL improvement. All patients reduced the intake of dopamine agonists, and five are currently free from L-Dopa medication, with no reported adverse events. We introduce a multicenter observational study to investigate whether early DBS treatment may affect the natural course of PD. Early application of DBS instead of L-Dopa administration could have a pathophysiological basis and be prompted by a significant incline on QoL through disease progression; however, the clinical value of this proposed paradigm shift should be addressed in clinical trials aimed at modulating the natural course of PD.


Interactions of 5-hydroxytryptamine and tricyclic antidepressants in the rat stomach strip and the guinea-pig ileum.
The interactions of the tricyclic antidepressants, amitriptyline and desipramine with 5-hydroxytryptamine (5-HT) were studied in the guinea-pig ileum and in the rat stomach strip. The tricyclics blocked the contractile response of the guinea-pig ileum and severely depressed the maximum [amitriptyline greater than desipramine (DMI)], however, they potentiated (DMI greater than amitriptyline) 5-HT-induced contractions of the RSS. The potentiation induced with DMI seems to be a combination of uptake inhibition and some post-receptor sensitization whilst amitriptyline-induced potentiation may be due to the former. Higher concentrations of the tricyclics contract the RSS. These contractions were absent in reserpinized RSS. Reserpinization reduced the potentiation after tricyclics and DMI was unable to enhance maximal concentrations of 5-HT, i.e. the post-receptor effect of DMI seems to have been abolished. It is concluded that whilst the tricyclics may be irreversible antagonists of 5-HT in the guinea-pig ileum, they potentiate the effect of 5-HT in the RSS by uptake inhibition, and, in the case of DMI, some additional post-receptor effect.


Inhibition of nitric oxide synthesis improves detoxication in inflammatory liver dysfunction in vivo.
Inflammatory stimulation of the liver induces nitric oxide (NO) biosynthesis and suppression of detoxication. In this study the effect of NO biosynthesis on cytochrome P-450 (CYP) enzyme activity was investigated by comparing in vivo and in vitro assays. To establish liver inflammation, CD rats were injected with Corynebacterium parvum (C. parvum) suspension. After 5 days NO biosynthesis was highly induced as indicated by increased NO2- plus NO3- serum concentrations. At the same time the aminopyrine breath test (ABT), measuring CYP activity in vivo, was reduced to 42% and the in vitro assay of aminopyrine turnover was suppressed to 12% of NaCl- injected controls. When C. parvum-injected animals were treated with the NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA), CYP activities significantly improved with an ABT of 76% and an in vitro aminopyrine turnover of 47% of controls. Neither C. parvum injections nor L-NMMA treatment resulted in a significant change of CYP protein concentrations. These data indicate that suppression of xenobiotic metabolism can be attenuated by inhibition of NO biosynthesis during an ongoing process of inflammation.


Effects of Long-term Antiepileptic Therapy on Carotid Artery Intima- Media Thickness.
Common first-line antiepileptic drugs (AEDs) used in children are valproate, phenytoin, and levetiracetam. Many side effects for these AEDs are reported including obesity, atherosclerosis, and metabolic syndrome. The aim of our study was to evaluate changes in carotid artery intima-media thickness (CIMT) in epileptic children and to correlate with lipid profile of those who are on long-term antiepileptic therapy. This case-control study was done over 18 months in department of pediatrics. Sample size was 84 with equal number of cases and controls. Epileptic children between 1 and 18 years of age receiving monotherapy with valproate, phenytoin, or levetiracetam for at least 6 months were included in the study. Measurement of CIMT was done by B mode ultrasonography. Lipid profile was analyzed. Statistical analysis was performed using one-way analysis of variance (ANOVA) test and Pearson correlation. Among 42 cases of epilepsy, 30 were on valproate, 9 on phenytoin, and 3 on levetiracetam monotherapy. No significant difference was noted in body mass index (BMI) among children receiving AED compared with that of controls (P = 0.82). Mean value for CIMT was significantly higher among valproate (0.43 ± 0.04, P ≤ 0.001), phenytoin (0.44 ± 0.04, P ≤ 0.001), and levetiracetam group (0.43 ± 0.03, P = 0.01) compared to controls (0.39 ± 0.01). Significant correlation was noted between CIMT and total cholesterol (P = 0.034), triglyceride (P = 0.011), low-density lipoprotein (P = 0.008), and very low-density lipoprotein (P = 0.011). Children on long-term monotherapy with valproate, phenytoin, and levetiracetam have significantly abnormal CIMT. This might be associated with atherosclerotic changes, and these children may require close follow-up to prevent cardiovascular and cerebrovascular risks.


In vitro binding analysis of legacy-linear and new generation-cyclic perfluoro-alkyl substances on sex hormone binding globulin and albumin, suggests low impact on serum hormone kinetics of testosterone.
In humans, serum testosterone (T) is largely bound to the sex hormone binding globulin (SHBG) and human serum albumin (hSA), resulting in a 2-3 % of unbound or "free" active quote (FT). Endocrine-disrupting chemicals, including perfluoro-alkyl substances (PFAS), are recognized to interfere with the hormonal axes, but the possible impact on the FT quote has not been addressed so far. Here we investigated the possible competition of two acknowledged PFAS molecules on T binding to SHBG and hSA. In particular, perfluoro-octanoic acid (PFOA) and acetic acid, 2,2-difluoro-2-((2,2,4,5-tetrafluoro-5(trifluoromethoxy)-1,3-dioxolan-4-yl)oxy)-ammonium salt (1:1) (C6O4) were used as, respectively, legacy-linear and new-generation-cyclic PFASs. Human recombinant SHBG 30-234 domain (SHBG30-234), produced in HEK293-F cells, and delipidated recombinant hSA were used as in vitro protein models. Isothermal Titration Calorimetry (ITC) and tryptophan fluorescence quencing (TFQ) were used to evaluate the binding modes of T and PFAS to SHBG30-234 and hSA. ITC revealed the binding of T to SHBG30-234 with a Kd of 44 ± 2 nM whilst both PFOA and C6O4 showed no binding activity. Results were confirmed by TFQ, since only T modified the fluorescence profile of SHBG30-234. In hSA, TFQ confirmed the binding of T on FA6 site of the protein. A similar binding mode was observed for PFOA but not for C6O4, as further verified by displacement experiments with T. Although both PFASs were previously shown to bind hSA, only PFOA is predicted to possibly compete with T for the binding to hSA. However, on the base of the binding stoichiometry and affinity of PFOA for hSA, this appears unlikely at the blood concentrations of the chemical documented to date.


[Study on the transcriptional activation of MTS1 gene beta promoter].
To investigate the effect of MTS1 gene beta promoter transcriptional activation in T-cell acute lymphoblastic leukemia (T-ALL) cell lines and identify the fragment with transcriptional activation. Seven pGL3 recombinant plasmids with the same 3'-end transcriptional start site but the different 5'sequences were constructed by gene recombinant technique and transfected into Jurkat cell line which is biallelic deletion of MTS1 gene by transient transfection. Luciferase report gene was detected to observe beta promoter transcriptional activation. Seven pGL3 recombinant plasmids containing different fragments of beta promoter were obtained, all of them showed transcriptional activation in Jurkat cell line. Among them, the 0.38 kb fragment cut by SacII-SacI is fundamental in transcription. MTS1 gene beta promoter can be activated in Jurkat cell line.


Immediate screening method for predicting the necessity of massive transfusions in trauma patients: a retrospective single-center study.
Hemostatic resuscitation might improve the survival of severely injured trauma patients. Our objective was to establish a simplified screening system for determining the necessity of massive transfusions (MT) at an early stage in trauma cases. We retrospectively analyzed the cases of trauma patients who had been transported to our institution between November 2011 and October 2013. Patients who were younger than 18 years of age or who were confirmed to have suffered a cardiac arrest at the scene or on arrival were excluded. MT were defined as transfusions involving the delivery of ≥10 units of red blood cell concentrate within the first 24 h after arrival. A total of 259 trauma patients were included in this study (males: 178, 69%). Their mean age was 49 ± 20, and their median injury severity score was 14.4. Thirty-three (13%) of the patients required MT. The presence of a shock index of ≥1, a base excess of ≤ -3 mmol/L, or a positive focused assessment of sonography for trauma (FAST) result was found to exhibit sensitivity and specificity values of 0.97 and 0.81, respectively, for predicting the necessity of MT. Furthermore, this method displayed an area under the receiver operating characteristic curve of 0.934 (95% confidence interval, 0.891-0.978), which indicated that it was highly accurate. Our screening method based on the shock index, base excess, and FAST result is a simple and useful way of predicting the necessity of MT early after trauma.


Validation and use of microdialysis for determination of pharmacokinetic properties of the chemotherapeutic agent mitomycin C - an experimental study.
Mitomycin C is a chemotherapeutic agent used in the treatment of peritoneal surface malignancies, administered as hyperthermic intraperitoneal chemotherapy after cytoreductive surgery. Pharmacokinetic studies have been based on analyses of blood, urine and abdominal perfusate, but actual tissue concentrations of the drug have never been determined. Microdialysis is an established method for continuous monitoring of low-molecular substances in tissues, and in the present study microdialysis of mitomycin C was studied in vitro and in vivo. Using in vitro microdialysis, relative recovery was determined when varying drug concentration, temperature and perfusion flow rate. In vivo microdialysis was performed in rats to verify long-term stability of relative recovery in four compartments (vein, peritoneum, extraperitoneal space and hind leg muscle). Subsequently, intravenous and intraperitoneal bolus infusion experiments were performed and pharmacokinetic parameters were calculated. In vitro, compatibility of mitomycin C and microdialysis equipment was demonstrated, and relative recovery was stable over an adequate concentration range, moderately increased by raising medium temperature and increased when flow rate was reduced, all according to theory. In vivo, stable relative recovery was observed over seven hours. Mitomycin C exhibited fast and even distribution in rat tissues, and equal bioavailability was achieved by intravenous and intraperitoneal infusion. The half-life of mitomycin C calculated after intravenous infusion was 40 minutes. Mitomycin C concentration can be reliable monitored in vivo using microdialysis, suggesting that this technique can be used in pharmacokinetic studies of this drug during hyperthermic intraperitoneal chemotherapy.


Improved Precision in Surface-Enhanced Raman Scattering Quantification of Analyte through Dual-Modality Multisite Sensing.
Accurate quantification of analyte using surface-enhanced Raman spectroscopy (SERS) is a desired, yet unfulfilled, ability that could enable a plethora of diagnostic- and defense-related applications. The major hurdles to overcome to achieve this goal are expensive manufacturing for highly ordered and reproducible substrates and low reproducibility of substrates produced through low cost methods. A technology that can set industry standards for manufacturing/processing of SERS substrates is still yet to be achieved. A dual-modality multisite sensing approach was developed, that overcomes the limitations experienced when fabricating bottom-up, reproducible, sensitive, and low-cost SERS substrates. Electrochemistry was combined with SERS for dual-modality sensing to improve precision by adding redundancy and encoding features, thus increasing measurement robustness and predictability. This technique works by calibrating SERS response with respect to active surface area, a parameter known to be proportional to charge, which can be estimated via electrochemical measurements. The dual-modality multisite measurement demonstrates at least 2.8× improvement in assay precision compared to the traditional single-site Raman measurements. The technique yields overall improved precision of measurement and is not limited to any particular SERS substrate or geometry, and thus it can be adapted and incorporated readily in any SERS sensing assay.


Transient population dynamics of mosquitoes during sterile male releases: modelling mating behaviour and perturbations of life history parameters.
The release of genetically-modified or sterile male mosquitoes offers a promising form of mosquito-transmitted pathogen control, but the insights derived from our understanding of male mosquito behaviour have not fully been incorporated into the design of such genetic control or sterile-male release methods. The importance of aspects of male life history and mating behaviour for sterile-male release programmes were investigated by projecting a stage-structured matrix model over time. An elasticity analysis of transient dynamics during sterile-male releases was performed to provide insight on which vector control methods are likely to be most synergistic. The results suggest that high mating competitiveness and mortality costs of released males are required before the sterile-release method becomes ineffective. Additionally, if released males suffer a mortality cost, older males should be released due to their increased mating capacity. If released males are of a homogenous size and size-assortative mating occurs in nature, this can lead to an increase in the abundance of large females and reduce the efficacy of the population-suppression effort. At a high level of size-assortative mating, the disease transmission potential of the vector population increases due to male releases, arguing for the release of a heterogeneously-sized male population. The female population was most sensitive to perturbations of density-dependent components of larval mortality and female survivorship and fecundity. These findings suggest source reduction might be a particularly effective complement to mosquito control based on the sterile insect technique (SIT). In order for SIT to realize its potential as a key component of an integrated vector-management strategy to control mosquito-transmitted pathogens, programme design of sterile-male release programmes must account for the ecology, behaviour and life history of mosquitoes. The model used here takes a step in this direction and can easily be modified to investigate additional aspects of mosquito behaviour or species-specific ecology.


Postoperative results after laparoscopic approach for treatment of large hiatal hernias: is mesh always needed? Is the addition of an antireflux procedure necessary?
Laparoscopic approach has been suggested as the definitive treatment for large hiatal hernias. Reinforcement of the hiatoplasty and the need to perform antireflux surgery is still undergoing discussion. The purpose of this study was to evaluate the postoperative results, with special emphasis on the recurrence rate and reflux after surgery comparing the use or not of mesh reinforcement. This prospective study included 81 patients with a complete evaluation through a clinical questionnaire, barium sulfate radiologic evaluation, endoscopy, manometry, and 24-hour intraesophageal pH monitoring before and after a hiatoplasty with an antireflux procedure. Mesh reinforcement was used in 23 patients. Postoperative complications occurred in 11 patients (13.6%), without mortality. Recurrent hernia was observed in 10 patients without mesh reinforcement (12.3%), whereas those with mesh reinforcement showed no hiatal hernia recurrence (P = 0.33). Normal resting lower esophageal sphincter pressure was obtained after fundoplication in 87.2% of patients, and abnormal acid reflux was observed in 12.8% of patients after surgery. In conclusion, mesh reinforcement in patients with large Type IV could prevent recurrent hiatal hernias, and an antireflux procedure must be performed in order to avoid postoperative acid reflux.


Adipose tissue n-6 fatty acids and acute myocardial infarction in a population consuming a diet high in polyunsaturated fatty acids.
The Jewish population of Israel consumes a diet rich in n-6 polyunsaturated fatty acids (PUFAs), principally linoleic acid. The consequences of this diet for ischemic heart disease (IHD) remain unclear. We assessed the association of adipose tissue n-6 fatty acids, which are derived entirely from the diet, with acute myocardial infarction (AMI). A total of 180 cases and 492 IHD-free controls aged 25-64 were included in a population-based case-control study of Jerusalem residents hospitalized with a first AMI. Diet was assessed by the use of a food-frequency questionnaire and adipose tissue fatty acids by gas chromatography of biopsy samples taken from subcutaneous gluteal tissue. The data were analyzed by multivariate logistic regression. Dietary PUFAs (: 10.1% of energy) correlated (r = 0.43, P < 0.001) with adipose tissue linoleic acid, which constituted 25.6% of storage fatty acids. High intakes of linoleic acid were not associated with excess risk of AMI (age- and sex-adjusted odds ratio for the third versus the first tertile: 0.96; 95% CI: 0.62, 1.48; NS). In contrast, arachidonic acid, the long chain n-6 derivative of linoleic acid, was positively associated with AMI (age- and sex-adjusted odds ratio: 2.12; 95% CI: 1.33, 3.36; P = 0.004). With multivariate adjustment, there was no evidence for an adverse association of linoleic acid with AMI, whereas the risk associated with arachidonic acid persisted, albeit attenuated. A very high linoleic acid intake does not appear to confer increased risk of nonfatal AMI. Nonetheless, the increased risk associated with arachidonic acid, a finding that requires confirmation, tempers an inference that diets rich in n-6 fatty acids are safe vis-à-vis coronary health.


A coarse-grained model of DNA with explicit solvation by water and ions.
Solvation by water and ions has been shown to be vitally important for biological molecules, yet fully atomistic simulations of large biomolecules remain a challenge due to their high computational cost. The effect of solvation is the most pronounced in polyelectrolytes, of which DNA is a paradigmatic example. Coarse-grained (CG) representations have been developed to model the essential physics of the DNA molecule, yet almost without exception, these models replace the water and ions by implicit solvation in order to significantly reduce the computational expense. This work introduces the first coarse-grained model of DNA solvated explicitly with water and ions. To this end, we combined two established CG models; the recently developed mW-ion model [DeMille, R. C.; Molinero, V. J. Chem. Phys. 2009, 131, 034107], which reproduces the structure of aqueous ionic solutions without electrostatic interactions, was coupled to the three-sites-per-nucleotide (3SPN) CG model of DNA [Knotts, T. A., IV; et al. J. Chem. Phys. 2007, 126, 084901]. Using atomistic simulations of d(CGCGAATTCGCG)(2) as a reference, we optimized the coarse-grained interactions between DNA and solvent to reproduce the solvation structure of water and ions around CG DNA. The resulting coarse-grained model of DNA explicitly solvated by ions and water (mW/3SPN-DNA) exhibits base-pair specificity and ion-condensation effects and it is 2 orders of magnitude computationally more efficient than atomistic models. We describe the parametrization strategy and offer insight into how other CG models may be combined with a coarse-grained solvent model such as mW-ion.


Living with COVID-19: emotions and health during the pandemic.
COVID-19 has led and still leads all nations to adopt virus containment plans, considering possible strategies that help to live with the virus. The present study aimed to investigate the practical, psychological and emotional aspects of living with COVID-19. A total of 250 young adults (86% female), who lived in north-western Italy, took part in an online photovoice study, in respect of social distancing. They participated in a group discussion following the method SHOWeD. The researchers analyzed the photographs along with comments and transcripts from the SHOWeD group discussions using the triangulation process of visual and textual data. The collected data were analyzed on the basis of grounded theory, with the support of the NVivo 12 software. From the analysis of the emotions concerning the psychological health of participants, 4 categories were identified. The results reveal benefits and emotional aspects that have stimulated the increase of individual and community empowerment. COVID-19 has led and still leads all nations to adopt virus containment plans, considering possible strategies that help to live with the virus. From the analysis of the results it emerged how the online photovoice was a tool that allowed on the one hand the sharing of experiences and emotions related to living with COVID-19 and on the other hand to maintain social ties despite physical distance.


The New Stories/New Cultures after-school enrichment program: a direct cultural intervention.
This article describes the organization, curriculum, and outcomes for New Stories/New Cultures, an activity-based program for after-school enrichment in five schools in the low-income neighborhood near a major American university. The program encourages students (70% Hispanic-American, 30% African-American) to experience themselves as producers of culture, not just as consumers. Its methods include (a) creative team use of video equipment and other expressive media and (b) lessons about media literacy (i.e., making critical choices about images and activities depicted in popular culture and commercials). Outcome measures with the cohort of fifth and sixth graders support the programs occupation-based philosophy. They show that students are more likely to experience themselves as building skills when engaged in activities that are both challenging and enjoyable. The students reported greatest engagement and enjoyment in activities that were creative, team-based, and involving media production. These same activities were correlated with increased self-esteem. The term direct cultural intervention is used to describe the application of occupational principles and critical perspectives to provide a population with conceptual tools and skills for interpreting and successfully navigating the social world.


RECONSTRUCTION OF THE HUMAN HIPPOCAMPUS IN 3D FROM HISTOLOGY AND HIGH-RESOLUTION EX-VIVO MRI.
In this paper, we present methods for the reconstruction of 3D histological volumes of the human hippocampal formation from histology slices. Inter-slice alignment is guided by a graph-theoretic approach that minimizes the impact of badly distorted slices. The reconstruction is refined by iterative affine and deformable co-registration with a high-resolution MRI of the postmortem tissue sample. We present an evaluation of reconstruction accuracy that is based on measures of similarity between boundaries drawn on both histology and MRI. Our methodology is currently being applied to an MRI atlas of the human hippocampal formation, in which atlas anatomical labels are derived from segmentation of reconstructed histology.


Embryo-Engineered Nonhuman Primate Models: Progress and Gap to Translational Medicine.
Animal models of human diseases are vital in better understanding the mechanism of pathogenesis and essential for evaluating and validating potential therapeutic interventions. As close relatives of humans, nonhuman primates (NHPs) play an increasingly indispensable role in advancing translational medicine research. In this review, we summarized the progress of NHP models generated by embryo engineering, analyzed their unique advantages in mimicking clinical patients, and discussed the remaining gap between basic research of NHP models to translational medicine.


Seroprevalence of Toxoplasma gondii in water buffaloes and cats in Guangxi, China.
Toxoplasma gondii is a pathogen that poses a serious threat to human health and causes significant economic losses to the global livestock industry. The prevalence of toxoplasmosis infection has been reported to be high in humans and animals around the world, but the occurrence of the disease has not yet been reported in water buffaloes in Guangxi Zhuang Autonomous Region, southern China. To understand the overall seroprevalence of T. gondii infection in Guangxi, a total of 1041 water buffalo and 114 cat serum samples were examined using an indirect enzyme-linked immunosorbent assay (I-ELISA). Of the 1041 water buffaloes analyzed, an overall seroprevalence of 52.9% (551/1041) was obtained, with year, season, and city location being significant factors affecting the rate of T. gondii infection in water buffaloes (P < 0.001). The results also revealed a high seroprevalence of 57% (65/114) in cats. Given that buffalo milk and meat products are vital food sources, these findings suggest that toxoplasmosis in water buffaloes may be a public health threat. This study provides the first T. gondii seroprevalence data in Guangxi, which could contribute to the prevention and control of toxoplasmosis in the region.


Probiotic effects on immunity and microbiome in HIV-1 discordant patients.
Some HIV-1 infected patients are unable to completely recover normal CD4+ T-cell (CD4+) counts after achieving HIV-1 suppression with combined Antiretroviral Therapy (cART), hence being classified as immuno-discordant. The human microbiome plays a crucial role in maintaining immune homeostasis and is a potential target towards immune reconstitution. RECOVER (NCT03542786) was a double-blind placebo-controlled clinical trial designed to evaluate if the novel probiotic i3.1 (AB-Biotics, Sant Cugat del Vallès, Spain) was able to improve immune reconstitution in HIV-1 infected immuno-discordant patients with stable cART and CD4+ counts <500 cells/mm3. The mixture consisted of two strains of L. plantarum and one of P. acidilactici, given with or without a fiber-based prebiotic. 71 patients were randomized 1:2:2 to Placebo, Probiotic or probiotic + prebiotic (Synbiotic), and were followed over 6 months + 3-month washout period, in which changes on systemic immune status and gut microbiome were evaluated. Primary endpoints were safety and tolerability of the investigational product. Secondary endpoints were changes on CD4+ and CD8+ T-cell (CD8+) counts, inflammation markers and faecal microbiome structure, defined by alpha diversity (Gene Richness), beta diversity (Bray-Curtis) and functional profile. Comparisons across/within groups were performed using standard/paired Wilcoxon test, respectively. Adverse event (AE) incidence was similar among groups (53%, 33%, and 55% in the Placebo, Probiotic and Synbiotic groups, respectively, the most common being grade 1 digestive AEs: flatulence, bloating and diarrhoea. Two grade 3 AEs were reported, all in the Synbiotic group: abdominal distension (possibly related) and malignant lung neoplasm (unrelated), and 1 grade 4 AE in the Placebo: hepatocarcinoma (unrelated). Synbiotic exposure was associated with a higher increase in CD4+/CD8+ T-cell (CD4/CD8) ratio at 6 months vs baseline (median=0.76(IQR=0.51) vs 0.72(0. 45), median change= 0.04(IQR=0.19), p = 0.03). At month 9, the Synbiotic group had a significant increase in CD4/CD8 ratio (0.827(0.55) vs 0.825(0.53), median change = 0.04(IQR=0.15), p= 0.02) relative to baseline, and higher CD4+ counts (447 (157) vs. 342(73) counts/ml, p = 0.03), and lower sCD14 values (2.16(0.67) vs 3.18(0.8), p = 0.008) than Placebo. No effect in immune parameters was observed in the Probiotic arm. None of the two interventions modified microbial gene richness (alpha diversity). However, intervention as categorical variable was associated with slight but significant effect on Bray-Curtis distance variance (Adonis R2 = 0.02, p = 0.005). Additionally, at month 6, Synbiotic intervention was associated with lower pathway abundances vs Placebo of Assimilatory Sulphate Reduction (8.79·10-6 (1.25·10-5) vs. 1.61·10-5 (2.77·10-5), p = 0.03) and biosynthesis of methionine (2.3·10-5 (3.17·10-5) vs. 4·10-5 (5.66·10-5), p = 0.03) and cysteine (1.83·10-5 (2.56·10-5) vs. 3.3·10-5 (4.62·10-5), p = 0.03). At month 6, probiotic detection in faeces was associated with significant decreases in C Reactive Protein (CRP) vs baseline (11.1(22) vs. 19.2(66), median change= -2.7 (13.2) ug/ml, p = 0.04) and lower IL-6 values (0.58(1.13) vs. 1.17(1.59) ug/ml, p = 0.02) when compared with samples with no detectable probiotic. No detection of the probiotic was associated with higher CD4/CD8 ratio at month 6 vs baseline (0.718(0.57) vs. 0.58(0.4), median change = 0.4(0.2), p = 0.02). After washout, probiotic non-detection was also associated with a significant increase in CD4+ counts (457(153) vs. 416(142), median change = 45(75), counts/ml, p = 0.005) and CD4/CD8 ratio (0.67(0.5) vs 0.59(0.49), median change = 0.04 (0.18), p = 0.02). A synbiotic intervention with L. plantarum and P. acidilactici was safe and led to small increases in CD4/CD8 ratio and minor reductions in sCD14 of uncertain clinical significance. A probiotic with the same composition was also safe but did not achieve any impact on immune parameters or faecal microbiome composition.


Factors associated with safe completion of Fédération Equestre Internationale eventing cross-country (2008-2018).
Equestrian eventing involves competing in three phases: dressage, jumping, and cross-country. Competitors are ranked by number of penalties accrued-with those who have fewer penalties ranked higher. Completing the cross-country phase with zero obstacle penalties is commonly referred to as 'running clear'. Understanding factors associated with running clear can help athletes plan strategically for success, while also helping governing bodies to refine qualification criteria for elite levels. This study was carried out to identify factors associated with running clear in the cross-country phase of Fédération Equestre Internationale (FEI) eventing. Retrospective cohort study of 107 348 horse starts worldwide in all FEI competitions between January 2008 and December 2018. Multivariable logistic regression models constructed stepwise using a bi-directional process. Two study cohorts were assessed: a complete cohort that met all inclusion requirements and separately, a cohort that included only horses starting at the level above their previous start. Sixteen factors were associated with running clear. Factors associated with increased likelihood of doing so included lower event level, lower dressage score earlier in the event, fewer recent FEI event starts, and more clear runs in their previous three FEI events. For horses that had stepped up an event level, 14 of these factors were still associated with running clear. Data available covered only FEI events, no national federation competitions were available for inclusion in horse histories. No prior veterinary information or data on training were available. This study provides a framework that allows stakeholders to potentially better understand the appropriate level of competition for any particular horse/rider combination, given the combination's recent history. This could provide an additional direct benefit in terms of safety by reducing the likelihood of a combination falling during cross-country.


Which indoor residual spraying insecticide best complements standard pyrethroid long-lasting insecticidal nets for improved control of pyrethroid resistant malaria vectors?
Where resources are available, non-pyrethroid IRS can be deployed to complement standard pyrethroid LLINs with the aim of achieving improved vector control and managing insecticide resistance. The impact of the combination may however depend on the type of IRS insecticide deployed. Studies comparing combinations of pyrethroid LLINs with different types of non-pyrethroid IRS products will be necessary for decision making. The efficacy of combining a standard pyrethroid LLIN (DuraNet®) with IRS insecticides from three chemical classes (bendiocarb, chlorfenapyr and pirimiphos-methyl CS) was evaluated in an experimental hut trial against wild pyrethroid-resistant Anopheles gambiae s.l. in Cové, Benin. The combinations were also compared to each intervention alone. WHO cylinder and CDC bottle bioassays were performed to assess susceptibility of the local An. gambiae s.l. vector population at the Cové hut site to insecticides used in the combinations. Susceptibility bioassays revealed that the vector population at Cové, was resistant to pyrethroids (<20% mortality) but susceptible to carbamates, chlorfenapyr and organophosphates (≥98% mortality). Mortality of wild free-flying pyrethroid resistant An. gambiae s.l. entering the hut with the untreated net control (4%) did not differ significantly from DuraNet® alone (8%, p = 0.169). Pirimiphos-methyl CS IRS induced the highest mortality both on its own (85%) and in combination with DuraNet® (81%). Mortality with the DuraNet® + chlorfenapyr IRS combination was significantly higher than each intervention alone (46% vs. 33% and 8%, p<0.05) demonstrating an additive effect. The DuraNet® + bendiocarb IRS combination induced significantly lower mortality compared to the other combinations (32%, p<0.05). Blood-feeding inhibition was very low with the IRS treatments alone (3-5%) but increased significantly when they were combined with DuraNet® (61% - 71%, p<0.05). Blood-feeding rates in the combinations were similar to the net alone. Adding bendiocarb IRS to DuraNet® induced significantly lower levels of mosquito feeding compared to adding chlorfenapyr IRS (28% vs. 37%, p = 0.015). Adding non-pyrethroid IRS to standard pyrethroid-only LLINs against a pyrethroid-resistant vector population which is susceptible to the IRS insecticide, can provide higher levels of vector mosquito control compared to the pyrethroid net alone or IRS alone. Adding pirimiphos-methyl CS IRS may provide substantial improvements in vector control while adding chlorfenapyr IRS can demonstrate an additive effect relative to both interventions alone. Adding bendiocarb IRS may show limited enhancements in vector control owing to its short residual effect.


Measuring masses of large biomolecules and bioparticles using mass spectrometric techniques.
Large biomolecules and bioparticles play a vital role in biology, chemistry, biomedical science and physics. Mass is a critical parameter for the characterization of large biomolecules and bioparticles. To achieve mass analysis, choosing a suitable ion source is the first step and the instruments for detecting ions, mass analyzers and detectors should also be considered. Abundant mass spectrometric techniques have been proposed to determine the masses of large biomolecules and bioparticles and these techniques can be divided into two categories. The first category measures the mass (or size) of intact particles, including single particle quadrupole ion trap mass spectrometry, cell mass spectrometry, charge detection mass spectrometry and differential mobility mass analysis; the second category aims to measure the mass and tandem mass of biomolecular ions, including quadrupole ion trap mass spectrometry, time-of-flight mass spectrometry, quadrupole orthogonal time-of-flight mass spectrometry and orbitrap mass spectrometry. Moreover, algorithms for the mass and stoichiometry assignment of electrospray mass spectra are developed to obtain accurate structure information and subunit combinations.


Monocyclic phenolic compounds stabilize human insulin and suppress its amorphous aggregation: In vitro and in vivo study.
Insulin is a small protein with 51 residues that mediates glucose uptake, and an interesting model for studying protein misfolding and aggregation. The aggregated forms of insulin undergo loss of activity and can provoke unwanted immune responses. Use of small molecules is considered to be an affordable method to counteract this aggregation process and stabilize insulin. In this study, aggregated forms of human recombinant insulin have been produced following exposure to high temperature. Aggregation process was followed over time by checking absorbance with spectrophotometry in presence and absence of various concentrations of small phenolic compounds including eugenol and epinephrine. Effects of these compounds on the structure and function of incubated insulin were evaluated by spectrofluorimetry, melting temperature (Tm) measurement and insulin tolerance test on Wistar rats. Formation of heat-induced insulin aggregation can be effectively inhibited by 1 mM eugenol and epinephrine and both compounds were found to preserve insulin activity to a considerable extent. In conclusion, simple aromatic compounds could be tailored to act as potent anti-aggregation compounds for insulin.


A comparison of the behavior of C57BL/6 and C57BL/10 mice.
Selection of an appropriate animal model is a crucial first step in many research programs. The C57BL/6 (B6) mouse is the most widely used inbred mouse strain in biomedical research; this is particularly so in behavioral studies. However, there are several C57BL substrains, all derived from common ancestors. C57BL/10 (B10) mice are superficially almost identical to B6 mice in appearance and behavior and widely used in inflammation and immunology research, yet rarely in behavioral studies. The present study assessed the comparability of behavioral results from these two strains, to determine whether they could be used interchangeably in future behavioral experiments. The results showed that the behavior of B6 mice clearly differed from that of B10 mice: in tests of cognition, species-typical behaviors, and motor coordination the B6 strain performed better. Consequently, B6 mice will probably remain the preferred choice for behavioral studies. Interpretation of results derived from the B10 strain should take into account its particular behavioral characteristics.


Multicomponent analysis of the pancreatic adenocarcinoma progression model using a pancreatic intraepithelial neoplasia tissue microarray.
A multistep model for pancreatic adenocarcinoma has been proposed recently. In this model, well-defined, noninvasive ductal lesions are recognized as precursors of invasive cancer and have been classified under the nomenclature of pancreatic intraepithelial neoplasia, or PanIN. Increasing evidence suggests that PanINs represent true neoplasms of the pancreatic ductal epithelium, accumulating histologic and genetic abnormalities in their progression toward invasive cancer. We have constructed a tissue microarray containing 55 PanIN lesions of all histologic grades in order to perform a multicomponent analysis of the pancreatic adenocarcinoma progression model. The protein products of 14 genes encompassing a variety of functional classes, such as tumor suppressor genes (p53, Smad4/Dpc4), oncogenes (beta-catenin), cell cycle antigens (p16, cyclin D1), proliferation antigens (Ki-67, topoisomerase II alpha), and epithelial apomucins (MUC1, MUC2, MUC5), as well as "novel" genes described as differentially up-regulated in invasive pancreas cancer by global microarray expression analysis (mesothelin, prostate stem cell antigen, fascin, and 14-3-3varsigma), were analyzed by immunohistochemistry on the PanIN tissue microarray. Comparison of the results from the current study with previously published data performed on routine histologic sections of PanINs demonstrates that tissue microarrays are a valid platform for molecular analysis not only of invasive cancers but of precursor lesions as well. In addition, this study demonstrates that molecular abnormalities in PanINs are not random but can usually be stratified into "early" changes (e.g., expression of MUC5 and prostate stem antigen, or loss of p16), "intermediate" changes (e.g., expression of cyclin D1), and "late" changes (e.g., expression of p53, proliferation antigens, MUC1, mesothelin, and 14-3-3varsigma, or loss of Smad4/Dpc4). Understanding the molecular pathogenesis of precursor lesions of invasive pancreatic adenocarcinomas using a high-throughput tissue microarray-based approach is a valuable adjunct to designing rational strategies for early detection of this lethal neoplasm.


Morphology and composition of InAs/GaAs quantum dots.
Surface compositional maps of self-organized InAs/GaAs quantum dots were obtained with laterally resolved photoemission spectroscopy. We found a surface In concentration of about 0.85 at the center of the islands which decreases to 0.75 on the wetting layer. Comparison with concentration values found in the core of similar dots suggests a strong In segregation on the topmost surface layers of the dots and on the surrounding wetting layer. Furthermore, the morphological properties of the dots such as size and density have been measured with plan-view transmission electron microscopy and low energy electron microscopy.


How Cells Can Control Their Size by Pumping Ions.
The ability of all cells to set and regulate their size is a fundamental aspect of cellular physiology. It has been known for sometime but not widely so, that size stability in animal cells is dependent upon the operation of the sodium pump, through the so-called pump-leak mechanism (Tosteson and Hoffman, 1960). Impermeant molecules in cells establish an unstable osmotic condition, the Donnan effect, which is counteracted by the operation of the sodium pump, creating an asymmetry in the distribution of Na+ and K+ staving off water inundation. In this paper, which is in part a tutorial, I show how to model quantitatively the ion and water fluxes in a cell that determine the cell volume and membrane potential. The movement of water and ions is constrained by both osmotic and charge balance, and is driven by ion and voltage gradients and active ion transport. Transforming these constraints and forces into a set of coupled differential equations allows us to model how the ion distributions, volume and voltage change with time. I introduce an analytical solution to these equations that clarifies the influence of ion conductances, pump rates and water permeability in this multidimensional system. I show that the number of impermeant ions (x) and their average charge have a powerful influence on the distribution of ions and voltage in a cell. Moreover, I demonstrate that in a cell where the operation of active ion transport eliminates an osmotic gradient, the size of the cell is directly proportional to x. In addition, I use graphics to reveal how the physico-chemical constraints and chemical forces interact with one another in apportioning ions inside the cell. The form of model used here is applicable to all membrane systems, including mitochondria and bacteria, and I show how pumps other than the sodium pump can be used to stabilize cells. Cell biologists may think of electrophysiology as the exclusive domain of neuroscience, however the electrical effects of ion fluxes need to become an intimate part of cell biology if we are to understand a fundamental process like cell size regulation.


Blood lipid profiles and peripheral blood mononuclear cell cholesterol metabolism gene expression in patients with and without methotrexate treatment.
Methotrexate (MTX) is the most commonly prescribed disease-modifying antirheumatic drug (DMARD) in rheumatoid arthritis. ATP-binding cassette transporter-A1 (ABCA1) and 27-Hydroxylase (HY27) are known antiatherogenic proteins that promote cellular cholesterol efflux. In THP-1 macrophages, MTX can promote the reversal of cholesterol transport, limit foam cell formation and also reverse COX-2 inhibitor-mediated downregulation of ABCA1. Despite its antiatherogenic potential in vitro, the impact of clinical use of low-dose MTX on cholesterol metabolism in humans is unknown. Objective of the study was to examine whether clinical MTX use is associated with altered blood lipids and/or ABCA1/HY27 expressions. In all, 100 rheumatoid arthritis subjects were recruited from a medical center in central Taiwan. Plasma lipid profiles and peripheral blood mononuclear cell HY27 and ABCA1 expressions were compared between subjects taking MTX (MTX+) and other disease-modifying antirheumatic drugs (DMARDs) (MTX-). Dietary intake was assessed by a registered dietician. Though no difference observed in the blood lipids between MTX+ and MTX- subjects, the expressions of ABCA1 and HY27 were significantly elevated in MTX+ subjects (n = 67) compared to MTX- subjects (n = 32, p < 0.05). ABCA expression correlated with MTX doses (r = 0.205, p = 0.042), and MTX+ subjects are more likely to have increased HY27 compared to MTX- subjects (OR = 2.5, p = 0.038). Prevalence of dyslipidemia and overweight, and dietary fat/cholesterol intake were lower than that of the age-matched population. Although no differences were observed in the blood lipids, the potential impacts of MTX on cholesterol metabolism should not be overlooked and the atheroprotective effects from MTX induced HY27 and ABCA1 expressions may still be present in those persons with pre-existing dyslipidemia. We demonstrated novel findings on the increased gene expressions of atheroprotective protein HY27 and ABCA1 in human peripheral blood mononuclear cells (PBMCs) with clinical use of low-dose MTX. Whether MTX induced HY27 and ABCA1 expressions can protect against cardiovascular disease in patients with chronic inflammation through the facilitation of cholesterol export remains to be established. Further studies on the impacts of low-dose MTX on hypercholesterolemic patients are underway.


Evaluation of the antioxidant activity of Coreopsis tinctoria Nuff. and optimisation of isolation by response surface methodology.
The aim of the present study was to determine the antioxidant activity of Coreopsis tinctoria flowering tops (CTFT). Studies were conducted to obtain suitable extraction conditions for chlorogenic acid, quercetin, luteolin, apigenin and kaempferol, which were identified and quantified by HPLC. Response surface methodology was employed to optimise the ultrasound-assisted extraction conditions including extraction time, ethanol concentration and liquid-solid ratio. The antioxidant activity of the extracts was analysed using various antioxidant models, such as DPPH, ABTS and hydroxyl radical scavenging assay. CTFT extracted for 15.0 min with ethanol at a concentration of 60.4% and with liquidsolid ratio 27.5:1 possessed a considerable amounts of total flavonoids and polyphenols (18.9%). This extract showed higher scavenging activity of ABTS and hydroxyl radical activity than rutin, however not in the DPPH test. We may assume that CTFT possess antioxidant and free radical scavenging potentials.


Future trends of mesothelioma mortality in Japan based on a risk function.
Mesothelioma is a malignancy with poor prognosis. It is chiefly caused by asbestos exposure and its symptoms can occur about 30-50 yr after the initial exposure. This study aims to predict the future trends in mesothelioma mortality in Japan using a method that is an alternative to the age-cohort model. Our approach is based on a risk function that links mesothelioma mortality combined with data pertaining to the population, size of the labor force, and quantity of asbestos imports. We projected the number of deaths occurring in individuals aged 50-89 for yr 2003-2050 using risk functions. Our results have indicated that mesothelioma mortality among Japanese people aged 50-89 yr will continue to increase until 2027 and reach a maximum of 66,327 deaths in the years 2003-2050. Our estimate has also suggested that the number of mesothelioma deaths could be significantly reduced if there were adequate compliance with the administrative level guidelines for occupational asbestos exposure.


Contribution made by parabiosis to the understanding of energy balance regulation.
Parabiosis is a chronic preparation that allows exchange of whole blood between two animals. It has been used extensively to test for involvement of circulating factors in feedback regulation of physiological systems. The total blood volume of each animal exchanges approximately ten times each day, therefore, factors that are rapidly cleared from the circulation do not reach equilibrium across the parabiotic union whereas those with a long half-life achieve a uniform concentration and bioactivity in both members of a pair. Involvement of a circulating factor in the regulation of energy balance was first demonstrated when one member of a pair of parabiosed rats became hyperphagic and obese following bilateral lesioning of the ventromedial hypothalamus. The non-lesioned partner stopped eating, lost a large amount of weight and appeared to be responding to a circulating "satiety" factor released by the obese rat. These results were confirmed using different techniques to induce obesity in one member of a pair. Studies with phenotypically similar ob/ob obese and db/db diabetic mice indicated that the obese mouse lacked a circulating signal that regulated energy balance, whereas the diabetic mouse appeared insensitive to such a signal. Positional cloning studies identified leptin as the circulating factor and subsequent parabiosis studies confirmed leptin's ability to exchange effectively between parabionts. These studies also suggest the presence of additional unidentified factors that influence body composition. This article is part of a Special Issue entitled: Animal Models of Disease.


Bacteremia after tooth extractions studied with the aid of prereduced anaerobically sterilized culture media.
Both prereduced molten agar and broth and aerobic molten agar and broth were inoculated with blood samples collected from patients with periodontitis, but in otherwise good health, both before and after extraction of two or more teeth. Postoperative blood samples from 23 of 25 patients sampled yielded anaerobic and facultative species. Colony counts from nine samples yielded from less than 1 to over 100 colonies per ml of blood. Organisms detected were species belonging to the genera Bacteroides, Fusobacterium, Peptostreptococcus, Leptotrichia, Propionibacterium, Peptococcus, Veillonella, plus Streptococcus mitis, S. salivarius, vibrio forms, and strains resembling S. mutans. The data indicate that prereduced anaerobically sterilized culture medium with polyanethol sulfonate is effective for detecting anaerobic species in bacteremia and that anaerobic species can be prevalent in bacteremias immediately after tooth extraction in patients with periodontitis.


Stains on the caring mantle. Doctors in aboriginal Australia have a history.
Doctors new to Aboriginal Australia are not infrequently surprised that their choice to work with the needy and underprivileged is not cause for unconditional positive regard. The naïveté of this position reflects the assumption that the nature of medical work somehow separates doctors from other less caring Europeans who have intruded into Aboriginal Australia. In this paper the ambivalence with which medical professionals are viewed by Aborigines is explored through an examination of particular historical instances drawn from northern Australia, particularly the Kimberley region of Western Australia. It may be that a willingness to interrogate preconceived notions of privileged access will be in the best interests of the profession, and of doctors choosing to work in Aboriginal Australia.


Elucidation and characterization of oligonucleotide-accessible sites on HIV-2 leader region RNA.
The retroviruses, including the human pathogens HIV-1 and HIV-2, are diploid inasmuch as they encapsidate two copies of their RNA genome. Prior to or during encapsidation, two copies of full-length genomic RNA recognize and stably bind each other in a process called dimerization. RNA structures within the viral genome promote dimerization in both HIV-1 and HIV-2 and are located in the 5'-untranslated leader region. Inhibition of dimerization by mutation of these RNA signals has been demonstrated to drastically reduce viral infectivity and replication kinetics and, thus, represents a potential target for antiretroviral therapy. In this study, we identified sites in HIV-2 leader region RNA that are functionally accessible to hybridization with oligonucleotides (ODNs) by reverse transcription with random ODN libraries (RT-ROL). We then tested specific ODNs directed against these regions for their efficacy in inhibiting RNA dimerization in vitro. We determined that of several hybridization-competent ODNs, only two were very effective in inhibiting RNA dimerization. Both of these ODNs were complementary to viral RNA at the primer binding site (PBS). These results identify regions with high accessibility to ODN binding on HIV-2 RNA and help to map the region(s) essential for dimerization within the viral RNA.


Cognitive test performance predicts change in functional status at the population level: the MYHAT Project.
In the community at large, many older adults with minimal cognitive and functional impairment remain stable or improve over time, unlike patients in clinical research settings, who typically progress to dementia. Within a prospective population-based study, we identified neuropsychological tests predicting improvement or worsening over 1 year in cognitively driven everyday functioning as measured by Clinical Dementia Rating (CDR). Participants were 1682 adults aged 65+ and dementia-free at baseline. CDR change was modeled as a function of baseline test scores, adjusting for demographics. Among those with baseline CDR = 0.5, 29.8% improved to CDR = 0; they had significantly better baseline scores on most tests. In a stepwise multiple logistic regression model, tests which remained independently associated with subsequent CDR improvement were Category Fluency, a modified Token Test, and the sum of learning trials on Object Memory Evaluation. In contrast, only 7.1% with baseline CDR = 0 worsened to CDR = 0.5. They had significantly lower baseline scores on most tests. In multiple regression analyses, only the Mini-Mental State Examination, delayed memory for visual reproduction, and recall susceptible to proactive interference, were independently associated with CDR worsening. At the population level, changes in both directions are observable in functional status, with different neuropsychological measures predicting the direction of change.


Conformations of allyl amine: theory vs experiment.
The relative stabilities of the five conformers of allyl amine, a medium-size aliphatic molecule, were estimated by applying ab initio quantum mechanical methods at several levels of theory. The second-order Møller-Plesset perturbation method (MP2), quadratic configuration interaction including single and double excitations (QCISD), coupled-cluster with single and double excitations (CCSD) and CCSD plus perturbative triple excitations [CCSD(T)] were applied. The Dunning correlation consistent basis sets (through aug-cc-pVQZ and cc-pV5Z) were employed. The MP2 energies relative to the energy of the cis-trans conformer reported here appear to approach the basis set limit. The predicted allyl amine conformer energies approaching the Hartree-Fock basis set limit are 158 cm-1 (cis-gauche), -5 cm-1 (gauche-trans), and -146 cm-1 (gauche-gauche). The same three relative energies near the MP2 basis set limit are 135, 103, and 50 cm-1, respectively. The analogous energies deduced from experiment are 173 +/- 12, 92 +/- 8, and 122 +/- 5 cm-1. The theoretical results obtained in the present study suggest that satisfactory predictions of the conformer energetics of allyl amine may be achieved only by theoretical methods that incorporate consideration of correlation effects in conjunction with large basis sets. Evaluation of the zero-point vibrational energy corrections is critical, due to the very small classical energy differences between the five conformers of allyl amine. Agreement between theory and experiment for the gauche-gauche conformational energy remains problematical.


Photon subtracted squeezed states generated with periodically poled KTiOPO(4).
We present generation of photon-subtracted squeezed states at 860 nm, from nearly pure, continuous-wave squeezed vacua generated with a periodically-poled KTiOPO(4) crystal as a nonlinear medium of a subthreshold optical parametric oscillator. We observe various kinds of photon-subtracted squeezed states, including non-Gaussian states similar to the single-photon state and superposition states of coherent states, simply by changing the pump power. Nonclassicality of the generated states clearly shows up as its negative region around the origin of the phase-space distributions, i.e., the Wigner functions. We obtain the value, -0.083 at the origin of the Wigner function, which is largest ever observed without any correction for experimental imperfections.


[Visualization of laryngopharynx during swallowing of negative contrast media (air) with cine mode 64-row MDCT].
Video fluoroscopic examination of swallowing generally needs a contrast media such as a barium sulfate. Since the examination is usually performed in patients with dysphasia, there is a risk of aspiration. We tried to visualize the laryngopharynx during swallowing of negative contrast media (air) with 64-row multi-detector computed tomography (64-MDCT). Cine mode 64-MDCT was performed to visualize the laryngopharynx in 4 healthy volunteers during swallowing of negative contrast media (air). The data were converted to three-dimensional (3D) images of 2 conditions (air and bone) and sagittal images of the soft tissue condition at a workstation. These images were sent to a personal computer and modeled to 3D cine images with Digital Imaging and Communication in Medicine (DICOM) Viewer and Quick Time Player. 3D cine images demonstrated movements of the epiglottis, vallecula, piriform sinus, tongue, pharyngeal wall, hyoid bone and thyroid cartilage.


Suppression of monosodium urate crystal-induced acute inflammation by diets enriched with gamma-linolenic acid and eicosapentaenoic acid.
A subcutaneous air pouch formed in Sprague-Dawley rats was used to study the effect of diets enriched in gamma-linolenic acid (GLA) (in plant seed oil) and eicosapentaenoic acid (EPA) (in fish oil) on acute inflammation induced by monosodium urate crystals. The GLA-enriched diet suppressed significantly the cellular phase of inflammation (polymorphonuclear leukocyte accumulation, crystal phagocytosis, and lysosomal enzyme activity), but it had little effect on the fluid phase (exudate volume and protein concentration). In contrast, the EPA-enriched diet suppressed the fluid phase but not the cellular phase of inflammation. The findings indicate that the fluid and cellular phases of acute inflammation can be controlled independently. A combined diet of fish oil and plant seed oil (EPA-enriched and GLA-enriched) reduced both the cellular and fluid phases of inflammation. Thus, dietary provision of alternative substrates for oxidative metabolism (other than arachidonic acid) modifies monosodium urate crystal-induced acute inflammation.


Environmental and Human Health Impacts of Spreading Oil and Gas Wastewater on Roads.
Thirteen states in the United States allow the spreading of O&G wastewaters on roads for deicing or dust suppression. In this study, the potential environmental and human health impacts of this practice are evaluated. Analyses of O&G wastewaters spread on roads in the northeastern, U.S. show that these wastewaters have salt, radioactivity, and organic contaminant concentrations often many times above drinking water standards. Bioassays also indicated that these wastewaters contain organic micropollutants that affected signaling pathways consistent with xenobiotic metabolism and caused toxicity to aquatic organisms like Daphnia magna. The potential toxicity of these wastewaters is a concern as lab experiments demonstrated that nearly all of the metals from these wastewaters leach from roads after rain events, likely reaching ground and surface water. Release of a known carcinogen (e.g., radium) from roads treated with O&G wastewaters has been largely ignored. In Pennsylvania from 2008 to 2014, spreading O&G wastewater on roads released over 4 times more radium to the environment (320 millicuries) than O&G wastewater treatment facilities and 200 times more radium than spill events. Currently, state-by-state regulations do not require radium analyses prior to treating roads with O&G wastewaters. Methods for reducing the potential impacts of spreading O&G wastewaters on roads are discussed.


The relative benefits of tamoxifen in older women with T1 early-stage breast cancer treated with breast-conserving surgery and radiation therapy.
Small, hormone receptor-positive breast carcinomas in older women are associated with low local recurrence rates. The relative benefits of adjuvant hormonal therapy remain unclear in elderly women with small, node-negative breast cancer after breast-conserving surgery and adjuvant radiation therapy. From our institutional data base, 224 patients ≥65 years of age with T1N0M0 breast cancer treated with BCS+RT were identified. Of these, 102 patients (45.5%) received tamoxifen (TAM) and 122 patients (54.5%) did not (no-TAM). The median follow-up time was 62.6 months. The 10-year local relapse-free survival (LRFS) was 98% in both the TAM and no-TAM cohorts (p = 0.58); the 10-year DMFS was 83% TAM vs. 89% no-TAM (p = 0.91). There was no difference in 10-year contralateral breast relapse or overall survival (OS) between the two cohorts. In univariate and multivariate analysis, use of TAM was not associated with LRFS, distant metastases-free survival (DMFS), OS, or a reduction in contralateral breast cancers when compared with the no-TAM cohort. In this large cohort of T1N0 elderly breast cancer patients treated with CS+RT, the use of TAM did not appear to decrease ipsilateral breast relapse, contralateral breast relapse, distant metastasis, or OS.


[Intra-arterial digital subtraction angiography (i.a. DSA) in the preoperative diagnosis of abdominal masses].
The diagnostic value of i.a. DSA, compared with conventional arteriography (n = 48), was proven in patients (n = 116) with abdominal tumours including intrahepatic masses. Arteries supplying the tumour, anomalous vessels and pathological malignant vascular abnormalities were visualised reliably in i.a. DSA despite reduced spatial resolution. An improved definition of hypervascular tumours was possible during the early arterial phase and of hypovascular or avascular masses during the late parenchymal phase of i.a. DSA. The dynamic evaluation of contrast flow is simpler and more reliable using the continuous mode in i.a. DSA than with the static imaging of a conventional film sequence. The advantages of i.a. DSA in preoperative vascular mapping and in detailed diagnosis of abdominal tumours and masses result in replacing conventional arteriography with very few exceptions.