Validation of proposed DSM-5 criteria for autism spectrum disorder.
The primary aim of the present study was to evaluate the validity of proposed DSM-5 criteria for autism spectrum disorder (ASD). We analyzed symptoms from 14,744 siblings (8,911 ASD and 5,863 non-ASD) included in a national registry, the Interactive Autism Network. Youth 2 through 18 years of age were included if at least one child in the family was diagnosed with ASD. Caregivers reported symptoms using the Social Responsiveness Scale and the Social Communication Questionnaire. The structure of autism symptoms was examined using latent variable models that included categories, dimensions, or hybrid models specifying categories and subdimensions. Diagnostic efficiency statistics evaluated the proposed DSM-5 algorithm in identifying ASD. A hybrid model that included both a category (ASD versus non-ASD) and two symptom dimensions (social communication/interaction and restricted/repetitive behaviors) was more parsimonious than all other models and replicated across measures and subsamples. Empirical classifications from this hybrid model closely mirrored clinical ASD diagnoses (90% overlap), implying a broad ASD category distinct from non-ASD. DSM-5 criteria had superior specificity relative to DSM-IV-TR criteria (0.97 versus 0.86); however sensitivity was lower (0.81 versus 0.95). Relaxing DSM-5 criteria by requiring one less symptom criterion increased sensitivity (0.93 versus 0.81), with minimal reduction in specificity (0.95 versus 0.97). Results supported the validity of proposed DSM-5 criteria for ASD as provided in Phase I Field Trials criteria. Increased specificity of DSM-5 relative to DSM-IV-TR may reduce false positive diagnoses, a particularly relevant consideration for low base rate clinical settings. Phase II testing of DSM-5 should consider a relaxed algorithm, without which as many as 12% of ASD-affected individuals, particularly females, will be missed. Relaxed DSM-5 criteria may improve identification of ASD, decreasing societal costs through appropriate early diagnosis and maximizing intervention resources.


Cavity cooling a single charged levitated nanosphere.
Optomechanical cavity cooling of levitated objects offers the possibility for laboratory investigation of the macroscopic quantum behavior of systems that are largely decoupled from their environment. However, experimental progress has been hindered by particle loss mechanisms, which have prevented levitation and cavity cooling in a vacuum. We overcome this problem with a new type of hybrid electro-optical trap formed from a Paul trap within a single-mode optical cavity. We demonstrate a factor of 100 cavity cooling of 400 nm diameter silica spheres trapped in vacuum. This paves the way for ground-state cooling in a smaller, higher finesse cavity, as we show that a novel feature of the hybrid trap is that the optomechanical cooling becomes actively driven by the Paul trap, even for singly charged nanospheres.


Postirradiation erythropoietic recovery in splenectomized mice.
The influence of splenectomy on erythropoietic recovery of lethally X-irradiated mice injected with different doses of syngeneic bone marrow was studied. Splenectomized animals showed less activation than unoperated controls in the lower dose range; however, by increasing the number of injected cells the response obtained was similar in both groups. Providing an adequate number of stem cells is administered to the splenectomized recipients, an enhanced erythropoietic activity of the graft may compensate for the absence of the spleen, which is an important organ in postirradiation recovery. Suggested explanations for this observation are lack of an haematopoietic inhibitory effect of the irradiated spleen, or changes in the environment provided by the spleenless host.


Teenage women's use of contraceptives in two populations.
Adolescent patterns of contraceptive use might be different in various populations and might have changed in the last 30 years. More appropriate use of contraception could prevent unplanned pregnancy. We interviewed 378 women in rural East Tennessee and 396 women in suburban-urban Baltimore, all of whom were aged 18 to 50 years, in a convenience sampling about their memories of sexual experiences and early contraceptive use. First sexual experiences began at a younger age in women from Tennessee than in women from Baltimore. Sexual experiences occurred at an earlier age during the last 15 years in Baltimore women. Contraceptive use at first sexual experience has approximately doubled in both locations, from 7% to 15% in Tennessee and from 42% to 75% in Baltimore during the last 15 years. The primary reason in Baltimore is increased use of condoms. Condom use in Tennessee is very low. These two populations are using contraception and condoms differently. Although the two populations are diverse, it could help physicians to learn about the particular contraceptive practices of their patient population to help their patients more appropriately with their contraceptive needs.


Form deprivation myopia in mature common marmosets (Callithrix jacchus).
Experimental manipulations of visual experience are known to affect the growth of the eye and the development of refractive state in a variety of species including human and nonhuman primates. For example, it is well established that visual form deprivation causes elongation of the eye and myopia. The effects of such manipulations have generally been examined in neonatal or juvenile animals. Whether adolescent common marmosets (a new world primate) are susceptible to form deprivation myopia was studied. Five adolescent marmosets were used in this study. Monocular form deprivation was induced by lid closure for 12 to 20 weeks, starting between 299 and 315 days of age. The effects of deprivation were assessed with keratometry, A-scan ultrasonography, and cycloplegic refractions. Both eyes (treated and fellow control) were measured before lid-closure, at the end of the deprivation period, and several times over the following 8 to 12 weeks. Adolescent marmosets are susceptible to visual form deprivation myopia. The experimental eyes showed significant axial elongation and myopia relative to the fellow control eyes. These changes were smaller, however, than those observed in younger eyes deprived for comparable periods. Like juvenile animals, the adolescent marmosets did not show recovery from myopia over the period monitored. The period for susceptibility to form deprivation myopia in the marmoset monkey extends beyond the early developmental period when ocular growth is rapid and emmetropization normally takes place. Visual form deprivation in adolescent marmosets with adult-sized eyes results in increased ocular growth and myopia. These data suggest that visual factors may influence the growth and refractive development of the human eye after puberty and may be involved in late-onset myopia.


Informed consent in pragmatic trials: results from a survey of trials published 2014-2019.
To describe reporting of informed consent in pragmatic trials, justifications for waivers of consent and reporting of alternative approaches to standard written consent. To identify factors associated with (1) not reporting and (2) not obtaining consent. Survey of primary trial reports, published 2014-2019, identified using an electronic search filter for pragmatic trials implemented in MEDLINE, and registered in ClinicalTrials.gov. Among 1988 trials, 132 (6.6%) did not include a statement about participant consent, 1691 (85.0%) reported consent had been obtained, 139 (7.0%) reported a waiver and 26 (1.3%) reported consent for one aspect (eg, data collection) but a waiver for another (eg, intervention). Of the 165 trials reporting a waiver, 76 (46.1%) provided a justification. Few (53, 2.9%) explicitly reported use of alternative approaches to consent. In multivariable logistic regression analyses, lower journal impact factor (p=0.001) and cluster randomisation (p<0.0001) were significantly associated with not reporting on consent, while trial recency, cluster randomisation, higher-income country settings, health services research and explicit labelling as pragmatic were significantly associated with not obtaining consent (all p<0.0001). Not obtaining consent seems to be increasing and is associated with the use of cluster randomisation and pragmatic aims, but neither cluster randomisation nor pragmatism are currently accepted justifications for waivers of consent. Rather than considering either standard written informed consent or waivers of consent, researchers and research ethics committees could consider alternative consent approaches that may facilitate the conduct of pragmatic trials while preserving patient autonomy and the public's trust in research.


An intrinsically compressible and stretchable all-in-one configured supercapacitor.
An all-in-one configured supercapacitor which is intrinsically compressible and stretchable is fabricated via one-step deposition of electrocapacitive nanocomposites onto two sides of a compressible and stretchable polyacrylic acid (PAA) hydrogel electrolyte. The supercapacitor affords a large degree of compression (up to 60% strain) without affecting the electrochemical performance. The exact same device also sustains stretching up to 150% strain, and meanwhile effectively powers an LED bulb.


Progesterone in Periplaneta americana and Neobellieria bullata adults from the procuticle phase until first progeny production.
A significant amount of progesterone-like immunoreactive material (150 ng/g) was measured by EIA in the procuticle phase of adult of both sexes of Periplaneta americana. This peak markedly decreased to 1-10 ng/g during sclerotization and was unlikely to be of dietary origin. In the case of 0-hr-old P. americana adults 96-98% of progesterone-like material was localized in the digestive tract and Malpighian tubules. In contrast, a relatively low level of progesterone-like immunoreactive material was measured in 0-hr-old Neobellieria bullata adults. Activity of 3beta-HSD/isomerase converting pregnenolone to progesterone was high (22-43 fmol/mg protein/20 min) in 0-hr-old P. americana adults and significantly fell during sclerotization. High progesterone levels (13-16 ng/g), measured by HPLC-RIA, coexist with high levels of 3beta-HSD/isomerase activity. Orally active human contraceptives (ethisterone, ethynodiol, ethynodiol diacetate, lynestrenol, mestranol, norgestrel, norethynodrel, tamoxifen citrate, and mifepristone) which act on mammalian steroid receptors had no significant effects on progeny production in either polytrophic or meroistic insect ovaries even at concentration of 5000 mg/kg.


Attachment, proliferation and collagen type I mRNA expression of human gingival fibroblasts on different biodegradable membranes.
The purpose of this study was to investigate adhesion, proliferation and type I collagen (COL I) mRNA expression of gingival fibroblasts on different membranes used in periodontal applications. Collagen (C), acellular dermal matrix (ADM) and polylactic acid; polyglycolic acid; lactide/glycolide copolymer (PLGA) biodegradable membranes were combined with gingival fibroblasts in culture and incubated for 48 h. Cell adhesion was examined with scanning electron and confocal microscopy. MTT assay was used to measure proliferation. COL I mRNA expression was assessed using quantitative-polymerase chain reaction (QPCR). The PLGA group exhibited the lowest cell survival on day 5 and 10, and lowest cell proliferation on days 5, 10 and 14. While cell proliferation was similar in C and ADM groups, the C membrane showed a slightly greater increase in viable cells to day 10. Confocal and scanning electron microscopy confirmed the results of proliferation and MTT assays. The highest COL I mRNA expression was noted in the PLGA membrane group when compared to the C (p < 0.01) and ADM (p < 0.05) membrane groups. These data revealed that adherence and proliferation of primary gingival fibroblasts on collagen-based C and ADM membranes is better than that seen with PLGA membranes, and thus may be preferable in the treatment of gingival recession defects.


[Hyperbaric oxygenation with low excess pressure implemented in the pressure chamber within the complex intensive care for combined craniocerebral trauma].
An attempt was made to apply the hyperbaric oxygenation (HBO) in victims with craniocerebral trauma. HBO sessions were carried out in a low excess pressure (0.2 ATA) with the simultaneous use of clonidin as antihinoxant and antioxidant (2 mkg/kg), which was done for the purpose of eliminating or decreasing the pronouncement degree of additional stimulation of the lipid-peroxidation processes by hyperbaric oxygen (such processes were already stimulated by combined trauma). Requirements are substantiated on the need in a differential approach to the indication of the HBO course even if it is implemented in the described (sparing) regime. If there is a negative body response to the HBO effect, which happened in 8 of 18 persons according to our observations, the HBO course must be interrupted.


[Hepatitis B in children: natural history and therapy].
Children with chronic hepatitis B, face life long disease and complications of cirrhosis and hepatocarcinoma. Naturally, it is estimated that half to two third of the children will clear the hepatitis Be antigen during childhood. Treatments aim to increase the HBe Ag to Ab seroconversion rate, which may also favour the loss of HBs antigen, ultimate goal. Interferon alpha was the first approved treatment for pediatric chronic hepatitis B, and was shown to increase the HBe ag loss from 11% in control group to 26% in treated patients (5 MU/square meter body surface area for six months) at one year, and 33% at 18 months. Side effects include mainly fever, flu like symptoms, and growth impairment during the treatment phase. Nucleotide analogues have now emerged as a promising alternative to treat chronic hepatitis B. The optimal dose for children is established to 3 mg/kg once daily up to 12 years old. Efficacy trials show complete virologic response in 23% of all treated patients after one year, as compared to 13% in the placebo group, and in 34% of patients with basal transaminases above two times upper limit of normal; versus 16% in controls. Lamivudine inhibits viral DNA which favours cellular immune response. Lamivudine resistance due to variant viruses is observed in 19% of children after one year. Other nucleotide analogues, such as entecavir and adefovir will soon be tested in children, and combination with Lamivudine may improve results. Finally, vaccine technology is being tested in adults, to induce a cellular immune response towards hepatitis B antigens, but no clinical benefit has so far been established.


Biomaterials, fibrosis, and the use of drug delivery systems in future antifibrotic strategies.
All biomaterials, when implanted into the body, elicit an inflammatory response that evolves into fibrovascular tissue formation on and around the material. As a result, material scientists and tissue engineers should be concerned about host response to tissue-engineered constructs that have a biomaterial component, because the host response to this component will interfere with device function and reduce the lifespan of tissue engineering devices in vivo. The fibrotic response to biomaterials is not unlike pathological fibrosis of the liver, lung, kidney, and peritoneum in many ways: i) the presence of mononuclear leukocytes are common in the local environment of both pathological fibrosis and biomaterial-induced fibrosis even though cells of mesenchymal origin are responsible for laying the majority of the extracellular matrix; ii) paracrine-signaling molecules, such as transforming growth factor beta;1, are essential mediators of fibrosis, whether it is pathological or biomaterial induced; and iii) injury and/or the presence of foreign materials (including bacterial components, toxins, or man-made objects) are essential initiators for the development of the fibrotic response. This review discusses mechanisms and research methodology related to pathological fibrosis that is of interest to researchers focused on biomaterials. Potential research models for the study of fibrosis from the fields of biomaterials and drug delivery are also discussed, which may be of interest to scientists working on the pathology of fibrotic disease.


Antihypertensive drugs and blood lipids.
Hypertension frequently coexists with hyperlipidaemia and it has been suggested that the potential benefits of blood pressure reduction may be compromised if lipid levels are not concurrently reduced. In addition, conventional first line antihypertensive drugs (thiazide diuretics and beta-blockers) produce adverse changes in blood lipids which are most apparent in the short-term but do not entirely disappear during chronic treatment. Of the alternative first-line antihypertensive agents, the calcium antagonist and ACE inhibitor drugs are lipid 'neutral' but only the alpha 1-blockers have been associated with favourable effects on the lipid profile.


Hepatoprotective potential of ethanolic extract of Pandanus odoratissimus root against paracetamol-induced hepatotoxicity in rats.
Pandanus odoratissimus (Pandanaceae) is popular in the indigenous system of medicines like Ayurveda, Siddha, Unani and Homoeopathy. In the traditional system of medicine various plant parts such as leaves, root, flowers, and oils are used as anthelmintic, tonic, stomachic, digestive and in the treatment of jaundice and various liver disorders. The aim was to investigate the hepatoprotective activity of ethanolic extract of the root of P. odoratissimus against paracetamol (PCM) induced hepatotoxicity in rats. Hepatotoxicity was induced in male Wistar rat by PCM (2 g/kg b.w. p.o. for 7 days). The ethanolic extract of P. odoratissimus root was administered at the dose level of 200 mg/kg and 400 mg/kg b.w. orally for 7 days and silymarin (100 mg/kg b.w. p.o.) as standard drug was administered once daily for a week. The hepatoprotective effect of ethanolic extract was evaluated by assessment of biochemical parameters such as serum glutamic oxaloacetic transaminase, serum glutamic-pyruvic transaminase, serum alkaline phosphatase, total and direct bilirubin and triglycerides. Histopathological study of rat liver was also done. Experimental findings revealed that the extract at dose level of 200 mg/kg and 400 mg/kg of b.w. showed dose dependant hepatoprotective effect against PCM induced hepatotoxicity by significantly restoring the levels of serum enzymes to normal that was comparable to that of silymarin, but the extract at dose level of 400 mg/kg was found to be more potent when compared to that of 200 mg/kg. Besides, the results obtained from histopathological study also support the study. From the results, it can be concluded that ethanolic extract of the root of P. odoratissimus afforded significant protection against PCM induced hepatotoxicity in rats.


Peritoneal Dialysis Patient Outcomes under the Medicare Expanded Dialysis Prospective Payment System.
Peritoneal dialysis (PD) use increased in the United States with the introduction of a new Medicare prospective payment system in January 2011 that likely reduced financial disincentives for facility use of this home therapy. The expansion of PD to a broader population and facilities having less PD experience may have implications for patient outcomes. We assessed the impact of PD expansion on PD discontinuation and patient mortality. A prospective cohort study was conducted of patients treated with PD at 90 days of ESKD. Patients were grouped by study start date relative to the Medicare payment reform: prereform (July 1, 2008 to December 31, 2009; n=10,585), interim (January 1, 2010 to December 31, 2010; n=7832), and reform period (January 1, 2011 to December 31, 2012; n=18,742). Patient characteristics and facility PD experience were compared at baseline (day 91 of ESKD). Patients were followed for 3 years for the major outcomes of PD discontinuation and mortality using Cox proportional hazards models. Patient characteristics, including age, sex, race, ethnicity, rurality, cause of ESKD, and comorbidity, were similar or showed small changes across the three study periods. There was an increasing tendency for patients on PD to be treated in facilities with less PD experience (from 34% during the prereform period being treated in facilities averaging <14 patients on PD per year to 44% in the reform period). Patients treated in facilities with less PD experience had a higher rate of PD discontinuation than patients treated in facilities with the most experience (hazard ratio [HR], 1.16; 95% confidence interval [95% CI], 1.10 to 1.23 for the first versus fifth quintile of PD experience). Nevertheless, the risk of PD discontinuation fell during the late interim period (HR, 0.88; 95% CI, 0.82 to 0.95) and most of the reform period (from HR, 0.85; 95% CI, 0.79 to 0.91 to HR, 0.94; 95% CI, 0.87 to 1.01). Mortality risk was stable across the three study periods. In the context of expanding PD use and declining facility PD experience, the risk of PD discontinuation fell, and there was no adverse effect on mortality. This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2019_09_12_CJN01610219.mp3.


Anterior tarsal tunnel syndrome secondary to missed talus fracture: a case report.
The anterior tarsal tunnel syndrome (ATTS) has first been described by Kopell and Thompson in 1963. The anterior tarsal tunnel is formed by the fascia lining the inferior extensor retinaculum and talus as well as the navicular bone. Many ATTS cases with various etiologies have been reported since the first description. We report here an ATSS case resulting from a fibro-osseous structure that occurred after a missed talus fracture. The ATTS diagnosis can be made with a comprehensive clinical neurological examination and electrophysiological study. The treatment is based on the underlying etiology, while surgery is the most common treatment providing successful outcomes in the long term.


Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B.
We have developed positional scanning synthetic combinatorial libraries to define the substrate specificity of carboxydipeptidases. The library Abz-GXXZXK(Dnp)-OH, where Abz is ortho-aminobenzoic acid, K(Dnp) is N(epsilon)-2,4-dinitrophenyl-lysine with free carboxyl group, the Z position was successively occupied with 1 of 19 amino acids (cysteine was omitted), and X represents randomly incorporated residues, was assayed initially with human cathepsin B, and arginine was defined as one of the best residues at the P(1) position. To examine the selectivity of S(1)('), S(2), and S(3) subsites, the sublibraries Abz-GXXRZK(Dnp)-OH, Abz-GXZRXK(Dnp)-OH, and Abz-GZXRXK(Dnp)-OH were then synthesized. The peptide Abz-GIVRAK(Dnp)-OH, which contains the most favorable residues in the P(3)-P(1)(') positions identified by screening of the libraries with cathepsin B, was hydrolyzed by this enzyme with k(cat)/K(m)=7288 mM(-1)s(-1). This peptide is the most efficient substrate described for cathepsin B to this point, and it is highly selective for the enzyme among the lysosomal cysteine proteases.


Novel patient-derived xenograft and cell line models for therapeutic testing of pediatric liver cancer.
Pediatric liver cancer is a rare but serious disease whose incidence is rising, and for which the therapeutic options are limited. Development of more targeted, less toxic therapies is hindered by the lack of an experimental animal model that captures the heterogeneity and metastatic capability of these tumors. Here we established an orthotopic engraftment technique to model a series of patient-derived tumor xenograft (PDTX) from pediatric liver cancers of all major histologic subtypes: hepatoblastoma, hepatocellular cancer and hepatocellular malignant neoplasm. We utilized standard (immuno) staining methods for histological characterization, RNA sequencing for gene expression profiling and genome sequencing for identification of druggable targets. We also adapted stem cell culturing techniques to derive two new pediatric cancer cell lines from the xenografted mice. The patient-derived tumor xenografts recapitulated the histologic, genetic, and biological characteristics-including the metastatic behavior-of the corresponding primary tumors. Furthermore, the gene expression profiles of the two new liver cancer cell lines closely resemble those of the primary tumors. Targeted therapy of PDTX from an aggressive hepatocellular malignant neoplasm with the MEK1 inhibitor trametinib and pan-class I PI3 kinase inhibitor NVP-BKM120 resulted in significant growth inhibition, thus confirming this PDTX model as a valuable tool to study tumor biology and patient-specific therapeutic responses. The novel metastatic xenograft model and the isogenic xenograft-derived cell lines described in this study provide reliable tools for developing mutation- and patient-specific therapies for pediatric liver cancer. Pediatric liver cancer is a rare but serious disease and no experimental animal model currently captures the complexity and metastatic capability of these tumors. We have established a novel animal model using human tumor tissue that recapitulates the genetic and biological characteristics of this cancer. We demonstrate that our patient-derived animal model, as well as two new cell lines, are useful tools for experimental therapies.


Heat shock elicits production of sexual inducer in Volvox.
In the green alga Volvox carteri, heat shock had an unusual and adaptive effect mediated by induced production of a well-defined effector molecule. Females of this species normally reproduce asexually in the absence of a potent sexual inducer produced by mature sexual males, but they generated egg-bearing sexual daughters after a brief exposure to elevated temperatures. This response involved an "autoinduction" of sexuality, in which heat-shocked somatic cells made and released the sexual inducer, which then redirected development of the reproductive cells. Males, including a sterile mutant incapable of producing inducer in the usual manner, also produced the inducer in response to heat shock. The phenomenon probably is of significance in the wild, where Volvox reproduces asexually in temporary ponds in spring but becomes sexual and produces dormant, overwintering zygotes before the ponds dry up in the summer heat.


The effects of calcium on protein turnover in skeletal muscles of the rat.
Several experimental procedures were used to increase the intracellular concentration of Ca2+ and determine its effects on protein turnover in isolated extensor digitorum longus and soleus muscle. These methods included the use of ionophore A23187, caffeine, dibucaine, thymol and procaine, all agents known to induce the release of calcium by acting either on the sarcolemma and/or on the sarcoplasmic reticulum. Another approach involved varying the external concentration of Ca2+ in the media in which the muscles were incubated. The changes in muscle Ca2+ concentrations after exposure to the various calcium-releasing agents were in keeping with accepted modes of action of these agents on muscle membranes. The findings suggest that increasing the sarcoplasmic concentration of Ca2+ inhibits protein synthesis and enhances protein breakdown. These catabolic effects of Ca2+ are compared with the changes induced in muscle protein turnover after exposure to insulin or cyclic nucleotides, and in myopathic muscle and situations of work overload. Attention is also drawn to some of the difficulties involved in definitively implicating Ca2+ as a factor involved in the normal regulation of protein turnover.


Pilot study of automated bullet signature identification based on topography measurements and correlations.
A procedure for automated bullet signature identification is described based on topography measurements using confocal microscopy and correlation calculation. Automated search and retrieval systems are widely used for comparison of firearms evidence. In this study, 48 bullets fired from six different barrel manufacturers are classified into different groups based on the width class characteristic for each land engraved area of the bullets. Then the cross-correlation function is applied both for automatic selection of the effective correlation area, and for the extraction of a 2D bullet profile signature. Based on the cross-correlation maximum values, a list of top ranking candidates against a ballistics signature database of bullets fired from the same model firearm is developed. The correlation results show a 9.3% higher accuracy rate compared with a currently used commercial system based on optical reflection. This suggests that correlation results can be improved using the sequence of methods described here.


One Collision - Two Substituents: Gas-Phase Preparation of Xylenes under Single-Collision Conditions.
The fundamental reaction pathways to the simplest dialkylsubstituted aromatics - xylenes (C6H4(CH3)2) - in high-temperature combustion flames and in low-temperature extraterrestrial environments are still unknown, but critical to understand the chemistry and molecular mass growth processes in these extreme environments. Exploiting crossed molecular beam experiments augmented by state-of-the-art electronic structure and statistical calculations, this study uncovers a previously elusive, facile gas-phase synthesis of xylenes through an isomer-selective reaction of 1-propynyl (methylethynyl, CH3CC) with 2-methyl-1,3-butadiene (isoprene, C5H8). The reaction dynamics are driven by a barrierless addition of the radical to the diene moiety of 2-methyl-1,3-butadiene followed by extensive isomerization (hydrogen shifts, cyclization) prior to unimolecular decomposition accompanied by aromatization via atomic hydrogen loss. This overall exoergic reaction affords a preparation of xylenes not only in high-temperature environments such as in combustion flames and around circumstellar envelopes of carbon-rich Asymptotic Giant Branch (AGB) stars, but also in low-temperature cold molecular clouds (10 K) and in hydrocarbon-rich atmospheres of planets and their moons such as Triton and Titan. Our study established a hitherto unknown gas-phase route to xylenes and potentially more complex, disubstituted benzenes via a single collision event highlighting the significance of an alkyl-substituted ethynyl-mediated preparation of aromatic molecules in our Universe.


In vitro affinity maturation of a specificity-determining region-grafted humanized anticarcinoma antibody: isolation and characterization of minimally immunogenic high-affinity variants.
HuCC49V10 (V10), a humanized anticarcinoma monoclonal antibody (Ab) CC49, was generated by grafting only the specificity-determining regions (SDRs) of CC49 onto the variable light and variable heavy frameworks of the human Abs LEN and 21/28'CL, respectively. SDRs are those residues of the complementarity-determining regions that are most critical for antigen (Ag) binding. Compared with HuCC49, which was developed by conventional complementarity-determining region grafting, V10 has lower reactivity to the sera from patients who were previously given murine CC49 in clinical trials, although its Ag-binding affinity is 2-3-fold lower than that of HuCC49. To generate variants of V10 with higher Ag-binding affinity and lower sera reactivity, in vitro affinity maturation of V10 was carried out using phage display technique. A limited library of Fabs was generated by replacing some of the SDRs with all possible residues located at the corresponding positions in human Abs. The library was enriched, by several rounds of panning, in Fabs that have high affinity for the TAG-72 Ag. The clones encoding the best binders were expressed in insect cells as whole Abs that were purified and characterized. Competition radioimmunoassay and surface plasmon resonance measurements showed that two of the isolates, V14 and V15, have higher binding affinity than that of V10. In addition, the surface plasmon resonance analysis showed that the variants V14 and V15, compared with the parental V10, have lower reactivity to the anti-V region Abs using sera from patients who received murine CC49. The two isolates, V14 and V15, which show higher Ag-binding reactivity and lower sera reactivity than the parental V10 Ab, are potentially more useful clinical reagents. These results demonstrate that phage display can be used to isolate variants of an Ab that are potentially less immunogenic in patients than the parental Ab from which they are derived.


Suppression of Rayleigh-scattering-induced noise in OEOs.
Optoelectronic oscillators (OEOs) are hybrid RF-photonic devices that promise to be environmentally robust high-frequency RF sources with very low phase noise. Previously, we showed that Rayleigh-scattering-induced noise in optical fibers coupled with amplitude-to-phase noise conversion in photodetectors and amplifiers leads to fiber-length-dependent noise in OEOs. In this work, we report on two methods for the suppression of this fiber-length-dependent noise: altering the amplitude-dependent phase delay of the OEO loops and suppressing the Rayleigh-scattering-induced noise in optical fibers. We report a 20 dB reduction in the flicker phase noise of a 6 km OEO via these suppression techniques.


Iron metabolism and cell membranes. III. Iron-induced alterations in HeLa cells.
The morphologic characteristics of acute iron loading were studied in HeLa cells incubated in an iron-enriched Eagle's medium containing 500 mug/ml of iron. Chemical studies showed that ferritin synthesis was rapidly induced and the concentration of intracellular ferritin increased up to 72 hours. Closely coupled with an increase in HeLa cell ferritin was a marked decrease in the rate of cell multiplication. The significant ultrastructural findings of iron-induced HeLa cell injury are characterized by the appearance of both autophagic multivesicular and residual bodies over the first 72 hours of iron incubation. The prominence of multivesicular bodies was noted after only 4 hours' incubation, with iron and myelin figures first appearing after 6 hours. Thus, the partial arrest of cell multiplication was associated with an increase in cytoplasmic residual bodies containing iron and other debris. The distribution of intracellular ferritin within HeLa cells differs significantly from the distribution described previously in hepatic parenchymal cells. In HeLa cells, ferritin particles were confined to lysosomal vesicles and were not identified in cell sap, endoplasmic reticulum, or Golgi apparatus.


Metabolomics analysis and biomarker identification for brains of rats exposed subchronically to the mixtures of low-dose cadmium and chlorpyrifos.
Cadmium (Cd) and chlorpyrifos (CPF) are widespread harmful environmental pollutants with neurotoxicity to mammals. Although the exposure to Cd and CPF at the same time may pose a significant risk to human health, the subchronic combined neurotoxicity of these two chemicals at low levels in the brain is poorly understood. In this study, we treated rats with three doses (low, middle, and high) of Cd, CPF, or their mixture for 90 days. No obvious symptom was observed in the treated animals except those treated with high-dose CPF. Histological results showed that middle and high doses of the chemicals caused neuronal cell damage in brains. GC-MS-based metabonomics analysis revealed that energy and amino acid metabolism were disturbed in the brains of rats exposed to the two chemicals and their combinations even at low doses. We further identified the unique brain metabolite biomarkers for rats treated with Cd, CPF, or both. Two amino acids, tyrosine and l-leucine, were identified as the biomarkers for Cd and CPF treatment, respectively. In addition, a set of five unique biomarkers (1,2-propanediol-1-phosphate, d-gluconic acid, 9H-purine, serine, and 2-ketoisovaleric acid) was identified for the mixtures of Cd and CPF. Therefore, the metabolomics analysis is more sensitive than regular clinical observation and pathological examination for detecting the neurotoxicity of the individual and combined Cd and CPF at low levels. Overall, these results identified the unique biomarkers for Cd and CPF exposure, which provide new insights into the mechanism of their joint toxicity.


Programmatic Mapping: Providing Evidence for High Impact HIV Prevention Programs for Female Sex Workers.
Programmatic mapping (PM) is a rapid and efficient mechanism to develop size estimates of key populations including female sex workers (FSWs) and geolocate them at physical locations in a systematic and scientific manner. At the macro level, this information forms the basis for allocating program resources, setting performance targets, and assess coverage. At a micro level, PM data provide specific information on hot spots, estimates of FSWs at those spots, and hot spot typology and days and times of operation, all of which provides targeted service delivery strategies. This information can provide a reliable platform to plan HIV prevention and treatment services to considerable scale and intensity. Above all, the entire PM process requires deep involvement of FSWs, which increases community ownership of the data and can lead to an increased uptake of services. Despite a few limitations, the approach is versatile and can be used in varied country contexts to generate important information about sex work and its dynamics. In this paper, we describe experiences and lessons learned from using evidence generated from PM of FSWs in multiple countries to develop HIV prevention programs at scale.


Reliability of a multi-segment foot model in a neutral cushioning shoe during treadmill walking.
Detailed kinematics of the foot has been frequently reported on in the literature, specifically using various multi-segment foot models. It is important to identify the reliability of a multi-segment foot model in a population of mixed genders and activity levels, while walking in commonly used footwear. The main objective of this study was to investigate the between-day reliability and within-session variability of the Oxford Foot Model (OFM) while walking in a neutral cushioning shoe. A 7-camera Vicon motion capture system was used along with 29 passive reflective markers, placed on the participant to examine the multi-segment foot kinematics of the left foot using the OFM. Windows were cut in New Balance 840 shoes following reports from a previous investigation to maintain shoe integrity during testing. Two walking sessions on separate days were collected for 12 healthy participants, with an average total of 22 gait cycles per day. ICCs ranged from 0.020 to 0.964 for between-day reliability, and within-session ICC values ranged from 0.268 to 0.985. Between-day ICC values of the relative measures (excursion and range of motion (ROM)) were higher than the absolute angle measures (angle at foot strike and peak angle). Largest differences were measured in the transverse plane, and the smallest differences in the sagittal plane. Bland-Altman plots revealed best agreement in the frontal and sagittal planes. SEM values ranged from 0.04 to 3.5 for the between-day reliability. Between-day reliability and within-session variability were comparable to previous studies for adults walking barefoot and shod. This research demonstrates that the OFM can produce reliable data when applied to the assessment of a shod foot.


Vaccine-associated immune-mediated hemolytic anemia in the dog.
Vaccination has been incriminated as a trigger of immune-mediated hemolytic anemia (IMHA) in dogs and in people, but evidence to support this association is lacking. In a controlled retrospective study, idiopathic IMHA was identified in 58 dogs over a 27-month period. When compared with a randomly selected control group of 70 dogs (presented for reasons other than IMHA) over the same period, the distribution of cases versus time since vaccination was different (P < .05). Fifteen of the dogs (26%) had been vaccinated within 1 month (mean, 13 days; median, 14 days; range, 1 to 27 days) of developing IMHA (P < .0001), whereas in the control group no marked increase in frequency of presentation was seen in the first month after vaccination. The dogs with IMHA were divided into 2 groups based on time since vaccination: the vaccine IMHA group included dogs vaccinated within 1 month of developing IMHA; the nonvaccine IMHA group included dogs that developed IMHA more than 1 month after vaccination. The recently vaccinated dogs with IMHA (vaccine IMHA group) had significantly lower platelet counts (P < .05) and a trend towards increased prevalence of intravascular hemolysis and autoagglutination when compared with the nonvaccine IMHA group. Similar mortality rates were seen in teh vaccine IMHA group (60%) and the nonvaccine IMHA group (44%), with the majority of fatalities (> 75%) occurring in the first 3 weeks after presentation. Persistent autoagglutination was a negative prognostic indicator for survival in both groups (P < .05). Presence of icterus and hyperbilirubinemia were negative prognostic indicators for survival in the nonvaccine IMHA group (P < .0001 and P < .01, respectively) but not in the vaccine IMHA group. In the recently vaccinated dogs, combination vaccines from various manufacturers against canine distemper, adenovirus type 2, leptospirosis, parainfluenza, and parvovirus (DHLPP) were involved in each case. Vaccines against rabies virus, Bordetella spp, coronavirus, and Lyme Borrelia were administrated concomitantly to some dogs. This study provides the first clinical evidence for a temporal relationship of vaccine-associated IMHA in the dog.


Influence of angioscopic vein graft preparation on development of neointimal hyperplasia in an organ culture model of human saphenous vein.
Angioscopy for in situ vein graft preparation has been criticized on the basis that the trauma of instrumentation may predispose to accelerated intimal hyperplasia, jeopardizing patency rates following infrainguinal revascularization. The aim of this study was to assess the effects of angioscopic preparation on endothelial integrity and smooth muscle cell (SMC) behavior in an established organ culture model of human saphenous vein (HSV). HSV was harvested from 12 patients during bypass surgery before and after angioscopic preparation. Endothelial integrity was evaluated by immunohistochemical staining with JC-70 and scanning electron microscopy (SEM); remaining segments of pre- and postangioscopy vein were maintained in culture for 14 days in medium supplemented with 30% fetal calf serum. Viability was confirmed by measurement of tissue adenosine triphosphate on day 14 and thickness of the neointima was measured by computerized image analysis of histologic sections. Monoclonal antibodies to proliferating cell nuclear antigen (PCNA) were used as an immunohistochemical marker for proliferating SMCs. There was a significant reduction in the percentage staining by JC-70 (71.3% versus 20.4%) in pre- versus postangioscopy vein (p = 0.002 by Wilcoxon's rank test; n = 12). This was supported by SEM images. Despite this, there were no significant differences between the pre- and postangioscopy HSVs after 14 days of culture with respect to neointimal thickness (61 versus 56 microns) and staining with PCNA (4.80 versus 4.08 nuclei per 10 microns), all according to Wilcoxon's rank test. Angioscopic vein graft preparation is associated with endothelial cell loss but does not induce additional neointimal hyperplasia in HSV in vitro. These results suggest that angioscopic manipulation does not alter SMC behavior.


[Sjögren's syndrome].
We carry out an updated review of Sjögren's syndrome on occasion of the presentation of three new clinical cases whose oral manifestations led the patients to first seek help. Each patient was subjected to clinical exploration, together with complementary evaluations, aimed at establishing an early and precise diagnosis of the syndrome. We consider it important to adopt a series of standardized diagnostic criteria and revise treatment to maintain the orodental conditions as favorable as possible.


The Presence of Ancient Core Genes Reveals Endogenization from Diverse Viral Ancestors in Parasitoid Wasps.
The Ichneumonoidea (Ichneumonidae and Braconidae) is an incredibly diverse superfamily of parasitoid wasps that includes species that produce virus-like entities in their reproductive tracts to promote successful parasitism of host insects. Research on these entities has traditionally focused upon two viral genera Bracovirus (in Braconidae) and Ichnovirus (in Ichneumonidae). These viruses are produced using genes known collectively as endogenous viral elements (EVEs) that represent historical, now heritable viral integration events in wasp genomes. Here, new genome sequence assemblies for 11 species and 6 publicly available genomes from the Ichneumonoidea were screened with the goal of identifying novel EVEs and characterizing the breadth of species in lineages with known EVEs. Exhaustive similarity searches combined with the identification of ancient core genes revealed sequences from both known and novel EVEs. One species harbored a novel, independently derived EVE related to a divergent large double-stranded DNA (dsDNA) virus that manipulates behavior in other hymenopteran species. Although bracovirus or ichnovirus EVEs were identified as expected in three species, the absence of ichnoviruses in several species suggests that they are independently derived and present in two younger, less widespread lineages than previously thought. Overall, this study presents a novel bioinformatic approach for EVE discovery in genomes and shows that three divergent virus families (nudiviruses, the ancestors of ichnoviruses, and Leptopilina boulardi Filamentous Virus-like viruses) are recurrently acquired as EVEs in parasitoid wasps. Virus acquisition in the parasitoid wasps is a common process that has occurred in many more than two lineages from a diverse range of arthropod-infecting dsDNA viruses.


Evaluation of High-Throughput SARS-CoV-2 Serological Assays in a Longitudinal Cohort of Patients with Mild COVID-19: Clinical Sensitivity, Specificity, and Association with Virus Neutralization Test.
The association between SARS-CoV-2 commercial serological assays and virus neutralization test (VNT) has been poorly explored in mild patients with COVID-19. 439 serum specimens were longitudinally collected from 76 healthcare workers with RT-PCR-confirmed COVID-19. The clinical sensitivity (determined weekly) of 9 commercial serological assays were evaluated. Clinical specificity was assessed using 69 pre-pandemic sera. Correlation, agreement, and concordance with the VNT were also assessed on a subset of 170 samples. Area under the ROC curve (AUC) was estimated at 2 neutralizing antibody titers. The Wantai Total Ab assay targeting the receptor binding domain (RBD) within the S protein presented the best sensitivity at different times during the course of disease. The clinical specificity was greater than 95% for all tests except for the Euroimmun IgA assay. The overall agreement with the presence of neutralizing antibodies ranged from 62.2% (95%CI; 56.0-68.1) for bioMérieux IgM to 91.2% (87.0-94.2) for Siemens. The lowest negative percent agreement (NPA) was found with the Wantai Total Ab assay (NPA 33% (21.1-48.3)). The NPA for other total Ab or IgG assays targeting the S or the RBD was 80.7% (66.7-89.7), 90.3% (78.1-96.1), and 96.8% (86.8-99.3) for Siemens, bioMérieux IgG, and DiaSorin, respectively. None of the commercial assays have sufficient performance to detect a neutralizing titer of 80 (AUC < 0.76). Although some assays show a better agreement with VNT than others, the present findings emphasize that commercialized serological tests, including those targeting the RBD, cannot substitute a VNT for the assessment of functional antibody response.


Cardiac manifestations of IgG4-related disease: a case series.
IgG4-related disease (IgG4-RD) is an autoimmune condition affecting almost every organ system, with an early inflammatory phase and later fibrotic consequences. Vascular manifestations, particularly, large-vessel involvement in IgG4-RD, are well described. However, important IgG4-related effects on medium-sized arteries and the pericardium are less well recognized. These less frequently reported cardiovascular effects of IgG4-RD include coronary artery stenosis, pericardial disease, cardiac masses, and valvular heart disease. This case series focuses on three patients that demonstrate the cardiovascular effects of IgG4-RD and the pitfalls and importance of early diagnosis. Cases 1 and 2 presented with cardiac manifestations prior to more typical organ systems being affected which led to a delay in diagnosis. Case 1 presented with an acute myocardial infarction secondary to IgG4-RD of the coronary arteries and Case 2 presented with pericarditis which progressed to pericardial constriction due to IgG4-RD. Case 3 already had a diagnosis of IgG4-RD from a prior renal biopsy which raised the index of suspicion that his pericardial disease and thoracic mass were also related to IgG4-RD. Cardiac manifestations of IgG4-RD remain under-recognized and include coronary artery and pericardial disease. These manifestations often precede more typical manifestations in other organ systems. Recognizing cardiac manifestations of IgG4-RD on cardiac imaging can raise clinical suspicion and act as a catalyst to ascertain a confirmatory diagnosis. Early diagnosis and treatment are crucial to prevent potentially fatal outcomes and irreversible fibrosis.


Anatomical changes in resection cavity during brain radiotherapy.
Brain tumors are in general treated with a maximal safe resection followed by radiotherapy of remaining tumor including the resection cavity (RC) and chemotherapy. Anatomical changes of the RC during radiotherapy can have impact on the coverage of the target volume. The aim of the current study was to quantify the potential changes of the RC and to identify risk factors for RC changes. Sixteen patients treated with pencil beam scanning proton therapy between October 2019 and April 2020 were retrospectively analyzed. The RC was delineated on pre-treatment computed tomography (CT) and magnetic resonance imaging, and weekly CT-scans during treatment. Isotropic expansions were applied to the pre-treatment RC (1-5 mm). The percentage of volume of the RC during treatment within the expanded pre-treatment volumes was quantified. Potential risk factors (volume of RC, time interval surgery-radiotherapy and relationship of RC to the ventricles) were evaluated using Spearman's rank correlation coefficient. The average variation in relative RC volume during treatment was 26.1% (SD 34.6%). An expansion of 4 mm was required to cover > 95% of the RC volume in > 90% of patients. There was a significant relationship between the absolute volume of the pre-treatment RC and the volume changes during treatment (Spearman's ρ = - 0.644; p = 0.007). RCs are dynamic after surgery. Potentially, an additional margin in brain cancer patients with an RC should be considered, to avoid insufficient target coverage. Future research on local recurrence patterns is recommended.


Allelic variants of the thiopurine S-methyltransferase deficiency in patients with ulcerative colitis and in healthy controls.
Thiopurine S-methyltransferase (TPMT) is a cytosolic enzyme that catalyzes the inactivation of mercaptopurine, azathioprine, and thioguanine. The genetic polymorphisms in the TPMT gene that regulate TPMT activity are inherited as an autosomal recessive trait and patients with genetically determined low levels of TPMT activity develop severe myelosuppression when treated with standard doses of the above-mentioned drugs. We have analyzed the frequencies of the allelic variants of the TPMT gene in a white European population of healthy blood donors from Spain and The Netherlands, and in a group of patients suffering from ulcerative colitis (UC) with a similar genetic background. Two hundred and thirteen unrelated healthy individuals (HC) and 146 UC patients were typed for the polymorphic sites at positions 460 (G-->A) and 719 (A-->G) of the TPMT gene using specific polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) methods. There were no significant differences between the allele frequencies observed in the group of UC patients and those of the control group (10% of cases were heterozygous carriers of a TPMT mutant allele). The most frequent mutant allele in both UC and HC groups was TPMT3A (A460-->G719) (60% of carriers). TPMT3B (A460-->A719) and TPMT3C (G460-->G719) alleles were more often found in our study than in previously reported studies, reflecting the different genetic backgrounds of the European populations analyzed. Genotyping methods provide a simple and reliable screening to identify patients with a high risk of developing severe bone marrow toxicity if treated with thiopurine drugs. In UC patients, TPMT genotype should be determined before the initiation of azathioprine therapy.


Medical Student Perceptions of Global Surgery at an Academic Institution: Identifying Gaps in Global Health Education.
Robust global health demands access to safe, affordable, timely surgical care for all. The long-term success of global surgery requires medical students to understand and engage with this emerging field. The authors characterized medical students' perceptions of surgical care relative to other fields within global health. An optional, anonymous survey was given to all Johns Hopkins medical students from February to March 2016 to assess perceptions of surgical care and its role in global health. Of 480 students, 365 (76%) completed the survey, with 150 (41%) reporting global health interests. One-third (34%) of responding students felt that surgical care is one of two fields with the greatest potential global health impact in the future, second to infectious disease (49%). A minority (28%) correctly identified that trauma results in more deaths worldwide than obstetric complications or HIV/AIDS, tuberculosis, and malaria combined. Relative to other examined fields, students perceived surgical care as the least preventive and cost-effective, and few students (3%) considered adequate surgical care the best indicator of a robust health care system. Students believed that practicing in a surgical field was least amenable to pursuing a global health career, citing several barriers. Medical students have several perceptions of global surgery that contradict current evidence and literature, which may have implications for their career choices. Opportunities to improve students' global health knowledge and awareness of global surgery career paths include updating curricula, fostering meaningful international academic opportunities, and creating centers of global surgery and global health consortia.


Cytogenetic and molecular study of the PRDX4 gene in a t(X;18)(p22;q23): a cautionary tale.
The PRDX4 gene located at Xp22 encodes for a member of the peroxiredoxin gene family. Genes within this family exhibit thioredoxin-dependent peroxidase activity and have been implicated in cellular functioning, including proliferation and differentiation. Recently, PRDX4 has been identified as a partner gene in a t(X;21) translocation in a patient with acute myeloid leukemia. To determine whether PRDX4 was involved in other translocations, leukemia cells from 15 patients with Xp22 abnormalities were screened for involvement of the gene using fluorescence in situ hybridization (FISH). One sample from a 41-year-old woman with acute lymphoblastic leukemia showed three signals when hybridized with the PRDX4 probe. Cytogenetic analysis of the sample had identified a t(X;18)(p22;q23). Assuming that the three signals indicated a break within the PRDX4 gene, we performed FISH experiments and successfully narrowed the breakpoint on chromosome 18 to a 50-kb region. Subsequent analysis using spectral karyotyping showed that the leukemic cells had undergone multiple rearrangements and that a third X chromosome was present, albeit rearranged. Additional FISH experiments revealed that the third PRDX4 signal was the result of a third copy of the gene. Analysis of the other rearrangements has helped to characterize the multiple abnormalities within the leukemic cells. The findings underscore the importance of using multiple techniques when analyzing complex chromosomal rearrangements in malignant cells.


[The structure of the aglycon of eremomycin--a new antibiotic of the polycyclic glycopeptide group].
Eremomycin is shown to be a new representative of the group of polycyclic glycopeptides. By the amino acid composition it is close to vancomycin but by the structure of triphenoxytriaminotricarboxylic acid it differs from vancomycin. Monodechlorovancomycinic acid was detected in eremomycin. On the basis of the data obtained in studies on the amino acid sequence and the molecule functional groups the structural formula of eremomycin aglycon was assigned. It is demonstrated that the chlorine-containing phenylserine fragment of monodechlorovancomycin acid is located in the N-end region of the aglycon peptide chain.


Advancing the science of symptom management.
Since the publication of the original Symptom Management Model (Larson et al. 1994), faculty and students at the University of California, San Francisco (UCSF) School of Nursing Centre for System Management have tested this model in research studies and expanded the model through collegial discussions and seminars. In this paper, we describe the evidence-based revised conceptual model, the three dimensions of the model, and the areas where further research is needed. The experience of symptoms, minor to severe, prompts millions of patients to visit their healthcare providers each year. Symptoms not only create distress, but also disrupt social functioning. The management of symptoms and their resulting outcomes often become the responsibility of the patient and his or her family members. Healthcare providers have difficulty developing symptom management strategies that can be applied across acute and home-care settings because few models of symptom management have been tested empirically. To date, the majority of research on symptoms was directed toward studying a single symptom, such as pain or fatigue, or toward evaluating associated symptoms, such as depression and sleep disturbance. While this approach has advanced our understanding of some symptoms, we offer a generic symptom management model to provide direction for selecting clinical interventions, informing research, and bridging an array of symptoms associated with a variety of diseases and conditions. Finally, a broadly-based symptom management model allows the integration of science from other fields.


Multiparametric monitoring of ischemia-reperfusion in rat kidney: effect of ischemic preconditioning.
Microelectrode technology is a promising tool for monitoring kidney ischemia and the changes induced by its therapeutic management. Ischemic preconditioning, that is, brief ischemic periods before sustained ischemia, has been shown to protect several organs, including the kidney, from ischemia-reperfusion injury. We tested whether the effect of preconditioning could be appraised by real-time measurement of parameters representative of tissue hypoxia. In a sample of pentobarbital-anesthetized and mechanically ventilated rats, we studied the effect of renal ischemic preconditioning (10-min ischemia and 10-min reflow interval) on subsequent ischemia-reperfusion (45 min and 60 min). Renal tissue electrical impedance, extracellular pH, and potassium concentration [K+] were measured continuously by implanted microelectrodes. Ischemia induced an early, rapid rise in extracellular potassium and impedance module, followed by a phase of slower increase, whereas pH decreased rapidly, reaching a plateau. Preconditioning treatment did not cause significant changes in interstitial pH and [K+] but increased ischemic tissue impedance. During reperfusion, the three variables recovered progressively; however, after a decline, electrical impedance showed a clear postischemic increase. This rise was suppressed by preconditioning. Real-time measurement of any of the three parameters showed capability for early detection of ischemia. In contrast with findings in myocardial tissue, preconditioning in the kidney did not increase potassium cell loss during ischemia or improve ischemic acidosis or tissue impedance. Electrical impedance increased for a second time during reperfusion, indicating the presence of a postischemic cellular edema; concealing this episode was the most noticeable effect of the preconditioning treatment.


Social groups and polarization of aesthetic values from symmetry and complexity.
When deciding what images we prefer, our brain must weigh many aesthetic variables, such as symmetry and complexity. To date, aesthetic research has mainly focused on investigating one variable at a time. In this article, we use symmetry and complexity to study the problem of multi aesthetic-variable interactions. For symmetry and complexity, there are two simple interaction hypotheses. The independence hypothesis proposes that the evaluation of aesthetic variables is mutually independent. Meanwhile, Birkhoff's aesthetic-measure hypothesis predicts that people prefer images high in symmetry and low in complexity, and dislike the opposite. To test these hypotheses, we generated images that systematically varied in levels of symmetry and complexity. We then compared the subjects' preference maps to identify regions of likes and dislikes. Unlike the predictions from these hypotheses, we found that most, but not all subjects, formed two distinct natural clusters, termed "islands," in terms of likes and dislikes. We also found that people with more art exposure were less likely to belong to an island. If someone did belong to an island, their gender influenced which cluster they belonged to. We discuss alternate hypotheses, possible mechanisms for the occurrence of islands, and their possible social implications.


[Disability and lymphedema].
Axillary lymph-nodal dissection (ALD) for the surgical treatment of breast cancer plays an important role in consideration above all of the incidence of early and late complications (upper limb edema, pain at thoracic wall, impaired function of the limb), that sometimes can hide the real benefit for the patient. Arm lymphedema due to breast cancer treatment particularly represents a significantly disabling complication, from both physical and psychological points of view. It is useful, therefore, to inform patients about the necessity of a preventive treatment aimed at improving prognosis in relation to a possible future disability. Authors report some clinical experiences of work groups who studied psycho-physical damage due to lymphedema after breast cancer treatment. Finally, therapeutic and preventive procedures are pointed out with the aim of reducing the psycho-physical burden of disability correlated to lymphedema.


[Proteolytic enzymes of microorganisms].
Microbial proteolytic enzymes are able to hydrolyze various protein substrates. Data concerning the ability to degrade proteins amongst various groups of microorganisms, such as bacteria, fungi, actinomycetes and yeasts, are collected in this article. The physical and chemical properties of proteolytic enzyme preparations and methods of their separation and purification have been examined. The possible ways of these enzymes application in industry and medicine have been discussed. The paper is presented in Ukrainian.


Reactive oxygen species from the uncoupling of human cytochrome P450 1B1 may contribute to the carcinogenicity of dioxin-like polychlorinated biphenyls.
Polychlorinated biphenyls (PCBs) are classified by the International Agency for Research on Cancer as probable human carcinogens. A subset of PCBs are described as 'dioxin like' because of similarities to 2,3,7,8-tetrachlorodibenzo-p-dioxin. Dioxin-like PCBs have been shown to tightly bind the active site of cytochrome P450 (CYP) 1A isoforms, primarily CYP1A1, resulting in inhibition of CYP activity and the generation of reactive oxygen species (ROS) as a result of uncoupling of the catalytic cycle. Human CYP1B1 (hCYP1B1) is an extrahepatic CYP closely related to hCYP1A1 and is overexpressed in the lungs of smokers. Moreover, hCYP1B1 has been found to be overexpressed in cancers derived from a number of tissue types, as well as in pre-malignant prostate tumours, implicating overexpression of hCYP1B1 as a risk factor for extrahepatic carcinogenesis. It has been demonstrated previously that hCYP1B1 is inhibited by dioxin-like PCBs, but whether or not it is uncoupled has not been investigated. In the current study, the ability of three dioxin-like PCBs 3,3',4,4'-tetrachlorobiphenyl, 3,3',4,4',5-pentachlorobiphenyl and 3,3',4,4',5,5'-hexachlorobiphenyl (PCB169) to inhibit hCYP1B1 and stimulate the formation of ROS in V79MZ cells (which lack endogenous CYPs) expressing hCYP1B1 was demonstrated. Moreover, the generation of ROS was also associated with increases in parameters of oxidative stress related to genotoxicity (DNA oxidation and lipid peroxidation). For PCB169, these effects were time and concentration dependent. These data identify a novel mechanism of genotoxicity for dioxin-like PCBs, as well as providing further evidence that overexpression of hCYP1B1 is a risk factor for extrahepatic carcinogenesis.


Qualitative and quantitative MALDI imaging of the positron emission tomography ligands raclopride (a D2 dopamine antagonist) and SCH 23390 (a D1 dopamine antagonist) in rat brain tissue sections using a solvent-free dry matrix application method.
The distributions of positron emission tomography (PET) ligands in rat brain tissue sections were analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI). The detection of the PET ligands was possible following the use of a solvent-free dry MALDI matrix application method employing finely ground dry α-cyano-4-hydroxycinnamic acid (CHCA). The D2 dopamine receptor antagonist 3,5-dichloro-N-{[(2S)-1-ethylpyrrolidin-2-yl]methyl}-2-hydroxy-6-methoxybenzamide (raclopride) and the D1 dopamine receptor antagonist 7-chloro-3-methyl-1-phenyl-1,2,4,5-tetrahydro-3-benzazepin-8-ol (SCH 23390) were both detected at decreasing abundance at increasing period postdosing. Confirmation of the compound identifications and distributions was achieved by a combination of mass-to-charge ratio accurate mass, isotope distribution, and MS/MS fragmentation imaging directly from tissue sections (performed using MALDI TOF/TOF, MALDI q-TOF, and 12T MALDI-FT-ICR mass spectrometers). Quantitative data was obtained by comparing signal abundances from tissues to those obtained from quantitation control spots of the target compound applied to adjacent vehicle control tissue sections (analyzed during the same experiment). Following a single intravenous dose of raclopride (7.5 mg/kg), an average tissue concentration of approximately 60 nM was detected compared to 15 nM when the drug was dosed at 2 mg/kg, indicating a linear response between dose and detected abundance. SCH 23390 was established to have an average tissue concentration of approximately 15 μM following a single intravenous dose at 5 mg/kg. Both target compounds were also detected in kidney tissue sections when employing the same MSI methodology. This study illustrates that a MSI may well be readily applied to PET ligand research development when using a solvent-free dry matrix coating.


Exercise modulates polarization of TAMs and expression of related immune checkpoints in mice with lung cancer.
Purpose: Many studies have found that both endurance exercise (EX) and high-intensity interval training (HIIT) have a positive therapeutic effect on the treatment of lung cancer patients, but the specific mechanism is unclear. Therefore, we investigated whether EX and HIIT could delay the progression of lung cancer by affecting the infiltration of tumor-associated macrophages (TAMs) and restoring the tumor phagocytic activity of TAMs in lung cancer tissue. Methods: BALB/c mice were divided into 4 groups. The mice were given saline as the saline group (Saline), and the mice were given urethane as the lung cancer mice. The lung cancer mice were randomly divided into the control group (CON), EX group, and HIIT group. After exercise, the cancer tissues were collected for RT-PCR, immunofluorescence staining, and Wes automated western blotting system analysis. Results: Compared with the Saline group, the mRNA levels of TAMs M1 markers IL-6, TNF-α, iNOS, and M2 markers CD206, IL-10, and Arg-1 in the CON group were significantly increased (P<0.05). There was no significant difference in the percentage of F4/80 positive cells among the groups. Compared with the CON group, the percentage of CD86-positive cells in TAMs in the EX group was significantly decreased (P<0.05). From the protein expression level, compared with the CON group, the expression of SIRPα in the EX group was significantly increased (P<0.0001) and the expression of PD-L1 had a tendency to increase (P=0.06). Compared with the CON group, the expressions of IL-10, IL-12, CD47, and CD24 in the HIIT group were significantly increased (P<0.05). In addition, compared with the CON group, plasma IFN-γ in the EX group and HIIT group was significantly increased (P<0.05). Conclusion: Lung cancer tissue presents an inflammatory tumor microenvironment. The therapeutic effect of exercise on lung cancer is independent of the infiltration of TAMs in lung cancer tissue. In addition, endurance exercise can reduce the proportion of M1-type TAMs in lung cancer tissues, while HIIT antagonistically regulates M1 and M2 polarization of TAMs by increasing the levels of IL-10 and IL-12 in lung cancer tissues and circulating IFN-γ. Finally, endurance exercise and HIIT can modulate the expression of some immune checkpoints in lung cancer tissues.


[A different approach in the treatment of aneurysms of the ascending aorta (author's transl)].
A safe method in therapy of the ascending aortic aneurysm by partial resection and reconstruction combined with reinforcement by a nylon net is described. Seven patients were operated upon without hospital mortality. Early operation is recommended, when there is only slight aortic insufficiency. In such a situation it is possible to reinforce the ascending aorta without ECC, to stabilize the progress of widening the ascending aorta and the aortic root.


Synthesis and biological evaluation of 1,4-naphthoquinones and quinoline-5,8-diones as antimalarial and schistosomicidal agents.
Improving the solubility of polysubstituted 1,4-naphthoquinone derivatives was achieved by introducing nitrogen in two different positions of the naphthoquinone core, at C-5 and at C-8 of menadione through a two-step, straightforward synthesis based on the regioselective hetero-Diels-Alder reaction. The antimalarial and the antischistosomal activities of these polysubstituted aza-1,4-naphthoquinone derivatives were evaluated and led to the selection of distinct compounds for antimalarial versus antischistosomal action. The Ag(II)-assisted oxidative radical decarboxylation of the phenyl acetic acids using AgNO(3) and ammonium peroxodisulfate was modified to generate the 3-picolinyl-menadione with improved pharmacokinetic parameters, high antimalarial effects and capacity to inhibit the formation of β-hematin.


An individual patient data meta-analysis on characteristics, treatments and outcomes of glioblastoma/ gliosarcoma patients with metastases outside of the central nervous system.
To determine the characteristics, treatments and outcomes of patients with glioblastoma multiforme (GBM) or gliosarcoma (GS) and metastases outside of the central nervous system (CNS). PubMed and Web of Science searches for peer-reviewed articles pertaining to GBM/ GS patients with metastatic dissemination were conducted using the keywords gliosarcoma, glioblastoma, GBM, metastasis, metastases and metastatic. Additionally, we performed hand search following the references from the selected papers. Cases with metastases to the CNS were excluded and evaluated in a separate study. 109 articles published between 1928 and 2013 were eligible. They reported on 150 patients. We observed a remarkable increase in the number of cases per decade over time. Median overall survival from diagnosis of metastasis (OSM+) was 6.0 ± 0.8 months and median overall survival from initial diagnosis (OSID) 13 ± 2.4 months. On univariate analyses, gender, age, the histological subtype, the time interval between initial diagnosis and diagnosis of metastasis and pulmonary involvement did not influence OSM+. We did not observe any substantial treatment progress. A comparison of the present cohort with 84 GBM/ GS patients with exclusive CNS dissemination suggests that metastases outside the CNS are related to a slightly more favorable outcome. The occurrence of extra-CNS metastasis from GBM/ GS is associated with a dismal prognosis, however it seems to compare slightly favorable to CNS dissemination. Crucial treatment progress has not been achieved over recent decades. A central registry should be considered to consecutively gain more information about the ideal therapeutic approach.


Evaluation of strategies for use of acellular pertussis vaccine in adolescents and adults: a cost-benefit analysis.
Pertussis is increasingly recognized as a source of infection in adults who then commonly infect young children. Immunity to illness caused by Bordetella pertussis is not long-lived, so optimal control of pertussis may require booster immunizations. In a cost-benefit analysis, we evaluated the benefits of 7 independent strategies for administering a pertussis booster, in the form of a diphtheria-tetanus-acellular pertussis vaccine, to adolescents and adults. Break-even vaccine costs for each strategy were calculated by dividing costs preventable by vaccine by the number of persons eligible for vaccination. Of these strategies, the most economical would be to immunize adolescents 10-19 years of age, which would prevent 0.7-1.8 million pertussis cases and save 0.6 dollars-1.6 billion dollars over a decade. Although justified by our analysis, routine adult booster vaccinations every decade would be more expensive and more difficult to implement. A recommendation for booster vaccinations every 10 years requires more information about duration of immunity, program costs, compliance, and nonmedical costs associated with pertussis.


A Cephalosporin Prochelator Inhibits New Delhi Metallo-β-lactamase 1 without Removing Zinc.
Antibacterial drug resistance is a rapidly growing clinical threat, partially due to expression of β-lactamase enzymes, which confer resistance to bacteria by hydrolyzing and inactivating β-lactam antibiotics. The increasing prevalence of metallo-β-lactamases poses a unique challenge, as currently available β-lactamase inhibitors target the active site of serine β-lactamases but are ineffective against the zinc-containing active sites of metallo-β-lactamases. There is an urgent need for metallo-β-lactamase inhibitors and antibiotics that circumvent resistance mediated by metallo-β-lactamases in order to extend the utility of existing β-lactam antibiotics for treating infection. Here we investigated the antibacterial chelator-releasing prodrug PcephPT (2-((((6R,7R)-2-carboxy-8-oxo-7-(2-phenylacetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)thio) pyridine 1-oxide) as an inhibitor of New Delhi metallo-β-lactamase 1 (NDM-1). PcephPT is an experimental compound that we have previously shown inhibits growth of β-lactamase-expressing E. coli using a mechanism that is dependent on both copper availability and β-lactamase expression. Here, we found that PcephPT, in addition to being a copper-dependent antibacterial compound, inhibits hydrolysis activity of purified NDM-1with an IC50 of 7.6 μM without removing zinc from the active site and restores activity of the carbapenem antibiotic meropenem against NDM-1-producing E. coli. This work demonstrates that targeting a metal-binding pharmacophore to β-lactamase-producing bacteria is a promising strategy for inhibition of both bacterial growth and metallo-β-lactamases.


Comparing microvascular outcomes at a large integrated health maintenance organization with flagship centers in the United States.
To determine if patients undergoing microvascular reconstructive head and neck surgery (MRHNS) at a large integrated health maintenance organization can expect outcomes similar to some of the best or flagship centers in the United States. Outcomes (flap loss, mortality, length of stay), eligibility (recent consecutive US center experience), high-experience (100 cases), high-volume (26 cases per year), and flagship criteria were prospectively defined. A systematic MEDLINE search identified 17 eligible reports. Independent, blinded medical reviewers identified 5 centers (29%) as flagship centers. The first 116 consecutive patients (average, 39 cases per year) who underwent MRHNS on this service. All 5 flagship centers are major academic health centers ranked in the top 18 "best head and neck hospitals" in the United States. Flap loss (1.7% vs 4.4% for flagship centers; range, 0.9%-8.8%) and mortality (2.6% vs 2.8% for flagship centers; range, 0.5%-6.3%) rates were not significantly different. Although lengths of stay in flagship centers were similar to each other and the literature (mean, 21.4 days; range, 20.1-22.5 days), our length of stay was significantly shorter (8.8 days, P<.001). For high-experience and high-volume centers, patients undergoing MRHNS at a large integrated health maintenance organization can expect morbidity and mortality outcomes similar to flagship centers in the United States, with shorter hospitalizations.


Mild form of oculocutaneous albinism type 1: phenotypic analysis of compound heterozygous patients with the R402Q variant of the TYR gene.
Oculocutaneous albinism type 1 (OCA1) is due to TYR mutations. c.1205G>A/p.Arg402Gln (R402Q) is a thermosensitive variant of the TYR gene that has been reported to be responsible for mild forms of OCA1. The aim of our study was to define the phenotype associated with this variant. In our retrospective series, among 268 patients diagnosed with OCA1, 122 (45.5%) harboured one pathogenic variant of TYR, and the R402Q variant ensured to be in trans by segregation analysis in 69 patients (25.7%), constituting the 'R402Q-OCA1' group. 146 patients harboured two pathogenic variants of the TYR gene other than R402Q. Clinical records were available for 119 of them, constituting the 'Classical-OCA1' group. Most R402Q-OCA1 patients presented with white or yellow-white hair at birth (71.43%), blond hair later (46.97%), a light phototype but with residual pigmentation (69.64%), and blue eyes (76.56%). Their pigmentation was significantly higher than in the classical-OCA1 group. All patients from the R402Q-OCA1 group presented with ocular features of albinism. However the prevalence of photophobia (78.13%) and iris transillumination (83.87%) and the severity scores of iris transillumination, retinal hypopigmentation and foveal hypoplasia were lower in the R402Q-OCA1 group. Visual acuity was higher in the R402Q-OCA1 group (0.38±0.21 logarithm of the minimum angle of resolution vs 0.76±0.24). Investigations concerning a possible additive effect of the c.575C>A/p.Ser192 (S192Y) variant of TYR in cis with R402Q, suggested by others, showed no significant impact on the phenotype. The R402Q variant leads to variable but generally mild forms of albinism whose less typical presentation may lead to underdiagnosis.


[Characteristics of thyroid para-follicular cells in the offspring of female rats with chronic experimental liver injury at different stages of postnatal ontogenesis].
Structural and functional features of thyroid gland parafol-licular cell (PFC) population were studied the in 48 animals born to females with chronic experimental liver injury induced by injection of D-galactosamine hydrochloride. Using Grimelius silver nitrate impregnation method, PFC numbers were calculated and their subtypes according to their granular content and degranulation degree determined. Compared with the animals of the control group, the increase of PFC number per unit area together with the number of degranulated cells and the cells with a predominant location of granules on vascular pole was found. At the same time, the increase in the number of cells with moderate and significant degranulation was detected. The changes observed may be indicative of increased functional activity of PFC population in the experimental animals


Individual sample conditioning in flow analysis. Determination of N-total in plant materials.
A flow-batch system allowing in-line individual sample matrix matching is proposed for analysis of sample lots with high variability in acidity. The feasibility of the approach is demonstrated in the spectrophotometric determination of total nitrogen in Kjeldahl digests, using a column with a slightly soluble reagent (AgCl). The solutions are sequentially injected by means of an 8-port selecting valve and processed in a mixing chamber that is also used as a monitoring unit. The system yields reproducible results (r.s.d. usually < 2.5%) and the sampling rate is 14 samples/h. The analytical curve is linear within 1.00 and 6.00% N (dry basis), and the regression coefficient is > 0.999 (n = 6). Results are in agreement with certified values of standard reference materials and with results obtained by conductometry.


Prior history of head trauma in Parkinson's disease.
The purpose of this study was to evaluate the relationship between head trauma and Parkinson's disease (PD). A questionnaire was completed by 97 PD patients (mean age 68.6 years, duration of disease 6.4 years) and 64 spouses acting as controls (mean age 63 years). Thirty-one PD patients reported head trauma before onset of PD (mean 32.3 years earlier), whereas 11 controls reported head injury before completing the survey (mean 26 years earlier) (p less than 0.05). Twenty PD patients and five controls reported head injury associated with alteration or loss of consciousness (p less than 0.05). Injury occurred a mean of 37.7 years before onset of PD and 37.2 years before survey completion in the two groups, respectively. In both of these comparisons we adjusted the frequencies to eliminate sex differences between groups. Comparison of new frequencies showed differences that were not significant. However, a trend toward significance was observed when examining head trauma with alteration of consciousness. Other investigators have reported similar results indicating that previous head trauma may be a risk factor in the etiology of PD. However, recall bias might have an effect on the outcome of these studies and this is discussed. The relationship between head trauma and PD is still tenuous.


The First Year Inventory: a longitudinal follow-up of 12-month-old to 3-year-old children.
The First Year Inventory is a parent-report measure designed to identify 12-month-old infants at risk for autism spectrum disorder. First Year Inventory taps behaviors that indicate risk in the developmental domains of sensory-regulatory and social-communication functioning. This longitudinal study is a follow-up of 699 children at 3 years of age from a community sample whose parents completed the First Year Inventory when their children were 12 months old. Parents of all 699 children completed the Social Responsiveness Scale-Preschool version and the Developmental Concerns Questionnaire to determine age 3 developmental outcomes. In addition, children deemed at risk for autism spectrum disorder based on liberal cut points on the First Year Inventory, Social Responsiveness Scale-Preschool, and/or Developmental Concerns Questionnaire were invited for in-person diagnostic evaluations. We found 9 children who had a confirmed diagnosis of autism spectrum disorder from the sample of 699. Receiver operating characteristic analyses determined that a two-domain cutoff score yielded optimal classification of children: 31% of those meeting algorithm cutoffs had autism spectrum disorder and 85% had a developmental disability or concern by age 3. These results suggest that the First Year Inventory is a promising tool for identifying 12-month-old infants who are at risk for an eventual diagnosis of autism spectrum disorder.


Peer Communication in Online Mental Health Forums for Young People: Directional and Nondirectional Support.
The Internet has the potential to help young people by reducing the stigma associated with mental health and enabling young people to access services and professionals which they may not otherwise access. Online support can empower young people, help them develop new online friendships, share personal experiences, communicate with others who understand, provide information and emotional support, and most importantly help them feel less alone and normalize their experiences in the world. The aim of the research was to gain an understanding of how young people use an online forum for emotional and mental health issues. Specifically, the project examined what young people discuss and how they seek support on the forum (objective 1). Furthermore, it looked at how the young service users responded to posts to gain an understanding of how young people provided each other with peer-to-peer support (objective 2). Kooth is an online counseling service for young people aged 11-25 years and experiencing emotional and mental health problems. It is based in the United Kingdom and provides support that is anonymous, confidential, and free at the point of delivery. Kooth provided the researchers with all the online forum posts between a 2-year period, which resulted in a dataset of 622 initial posts and 3657 initial posts with responses. Thematic analysis was employed to elicit key themes from the dataset. The findings support the literature that online forums provide young people with both informational and emotional support around a wide array of topics. The findings from this large dataset also reveal that this informational or emotional support can be viewed as directive or nondirective. The nondirective approach refers to when young people provide others with support by sharing their own experiences. These posts do not include explicit advice to act in a particular way, but the sharing process is hoped to be of use to the poster. The directive approach, in contrast, involves individuals making an explicit suggestion of what they believe the poster should do. This study adds to the research exploring what young people discuss within online forums and provides insights into how these communications take place. Furthermore, it highlights the challenge that organizations may encounter in mediating support that is multidimensional in nature (informational-emotional, directive-nondirective).


IIb-RAD-sequencing coupled with random forest classification indicates regional population structuring and sex-specific differentiation in salmon lice (Lepeophtheirus salmonis).
The aquaculture industry has been dealing with salmon lice problems forming serious threats to salmonid farming. Several treatment approaches have been used to control the parasite. Treatment effectiveness must be optimized, and the systematic genetic differences between subpopulations must be studied to monitor louse species and enhance targeted control measures. We have used IIb-RAD sequencing in tandem with a random forest classification algorithm to detect the regional genetic structure of the Norwegian salmon lice and identify important markers for sex differentiation of this species. We identified 19,428 single nucleotide polymorphisms (SNPs) from 95 individuals of salmon lice. These SNPs, however, were not able to distinguish the differential structure of lice populations. Using the random forest algorithm, we selected 91 SNPs important for geographical classification and 14 SNPs important for sex classification. The geographically important SNP data substantially improved the genetic understanding of the population structure and classified regional demographic clusters along the Norwegian coast. We also uncovered SNP markers that could help determine the sex of the salmon louse. A large portion of the SNPs identified to be under directional selection was also ranked highly important by random forest. According to our findings, there is a regional population structure of salmon lice associated with the geographical location along the Norwegian coastline.


Drosophila SIN3 isoforms interact with distinct proteins and have unique biological functions.
The SIN3 corepressor serves as a scaffold for the assembly of histone deacetylase (HDAC) complexes. SIN3 and its associated HDAC have been shown to have critical roles in both development and the regulation of cell cycle progression. Although multiple SIN3 isoforms have been reported in simple to complex eukaryotic organisms, the mechanisms by which such isoforms regulate specific biological processes are still largely uncharacterized. To gain insight into how SIN3 isoform-specific function contributes to the growth and development of a metazoan organism, we have affinity-purified two SIN3 isoform-specific complexes, SIN3 187 and 220, from Drosophila S2 cells and embryos. We have identified a number of proteins common to the complexes, including the HDAC RPD3, as well as orthologs of several proteins known to have roles in regulating cell proliferation in other organisms. We additionally identified factors, including the histone demethylase little imaginal discs and histone-interacting protein p55, that exhibited a preferential interaction with the largest SIN3 isoform. Our experiments indicate that the isoforms are associated with distinct HDAC activity and are recruited to unique and shared sites along polytene chromosome arms. Furthermore, although expression of SIN3 220 can substitute for genetic loss of other isoforms, expression of SIN3 187 does not support Drosophila viability. Together our findings suggest that SIN3 isoforms serve distinct roles in transcriptional regulation by partnering with different histone-modifying enzymes.


Selection and analysis of an optimized anti-VEGF antibody: crystal structure of an affinity-matured Fab in complex with antigen.
The Fab portion of a humanized antibody (Fab-12; IgG form known as rhuMAb VEGF) to vascular endothelial growth factor (VEGF) has been affinity-matured through complementarity-determining region (CDR) mutation, followed by affinity selection using monovalent phage display. After stringent binding selections at 37 degrees C, with dissociation (off-rate) selection periods of several days, high affinity variants were isolated from CDR-H1, H2, and H3 libraries. Mutations were combined to obtain cumulatively tighter-binding variants. The final variant identified here, Y0317, contained six mutations from the parental antibody. In vitro cell-based assays show that four mutations yielded an improvement of about 100-fold in potency for inhibition of VEGF-dependent cell proliferation by this variant, consistent with the equilibrium binding constant determined from kinetics experiments at 37 degrees C. Using X-ray crystallography, we determined a high-resolution structure of the complex between VEGF and the affinity-matured Fab fragment. The overall features of the binding interface seen previously with wild-type are preserved, and many contact residues are maintained in precise alignment in the superimposed structures. However, locally, we see evidence for improved contacts between antibody and antigen, and two mutations result in increased van der Waals contact and improved hydrogen bonding. Site-directed mutants confirm that the most favorable improvements as judged by examination of the complex structure, in fact, have the greatest impact on free energy of binding. In general, the final antibody has improved affinity for several VEGF variants as compared with the parental antibody; however, some contact residues on VEGF differ in their contribution to the energetics of Fab binding. The results show that small changes even in a large protein-protein binding interface can have significant effects on the energetics of interaction.


Paternal familial twinning: hypothesis and genetic/medical implications.
The phenomenon of paternally dependent familial twinning has been known in human and animal genetics since the 1920s, but still remains without any theoretical explanation and is indeed a neglected field of inquiry. Over the last two decades investigations in reproduction biology have discovered the significant role of multiple paternally dependent errors in fertilization including androgenic triploidy and moles. We suggest the hypothesis that the fathers of twins in the relevant families carry gene variants that increase the probability of dispermy, diplospermy and male pronucleus heterochrony as well as involvement of two male pronuclei in the fertilization of two female meiotic products. Any resulting twins would be an exceptional intermediate between MZ and DZ twins - and might properly be described as "sesquizygotic" (SZ). Paternal familial twinning may also go together with infertility due to triploidy, moles and chimerism. The hypothesis: (i) places the curiosities of paternally derived twinning within the framework of current knowledge of reproductive genetics and verifiable phenomena; (ii) predicts the existence of families in which twinning is associated with reproductive abnormalities; (iii) predicts an occurrence in relevant families of the third and intermediate category of SZ twins. Families with paternal twinning may thus provide the natural selective system for the search of unusual cases of primary chimeras, the frequency of which is still unknown.


Vascular complications after percutaneous nephrolithotomy: 10 years of experience.
To provide a contemporary look at vascular complications after percutaneous nephrolithotomy (PNL) with access performed solely by a urologist using fluoroscopic guidance. A retrospective review of 2792 patients who had 3338 PNLs at Indiana University Health Methodist Hospital and Mayo Clinic Rochester was performed. Patients who experienced significant bleeding requiring diagnostic renal angiography and superselective embolization (SSE) were reviewed and compared with the overall database. There were 15 patients (16 renal units) requiring renal angiography and SSE (0.48%). Mean time from PNL to bleeding was 7 days (range, 1-15 days) and to SSE was 9.6 days (range, 2-18 days). Mean drop in hemoglobin was 5.3 g/dL (range, 2-9 g/dL). Transfusion was needed in 9 patients (60%). There were no differences between the vascular complications group and the uneventful PNL group in mean age (55.06 vs 52.2 years; P = .519), UTI history (40% vs 38%; P = .92), mean operative time (125.8 vs 102.47 minutes; P = .192), the need for multiple access (18.75% vs 18%; P = .939), and access location. The vascular complications group had a lower stone burden than the uneventful PNL group (stones > 2 cm; 43.7% vs 74.03%; P = .014). The incidence of vascular complications in this contemporary series is one of the lowest reported to date. At our centers, vascular bleeding complications appear to be a random and rare event after PNL as we were unable to identify any specific risk factors. Early SSE avoided the need for blood transfusion in many patients.


The Cole relaxation frequency as a parameter to identify cancer in breast tissue.
To correlate the Cole relaxation frequencies obtained from measurements of the electrical properties of breast tissue to the presence or absence of cancer. Four-lead impedance measurements were obtained on ex vivo specimens extracted during surgery from 187 volunteer patients. Data were acquired with a commercial Solartron impedance bridge employing 4-lead Ag-AgCl or blackened platinum (BPt) electrodes at frequencies logarithmically spaced from 1 Hz to 3.2 × 10(7) Hz utilizing 6-10 frequencies per decade. The Cole frequencies obtained from these measurements were correlated with the tissue health status (cancer or noncancer) obtained from histological analysis of the specimens. Analysis of the impedance measurements showed that the Cole relaxation frequencies correlated to the presence or absence of cancer in the examined tissue with a sensitivity up to 100% (95% CI, 99%-100%) and a specificity up to 85% (95% CI, 79%-91%) based on the ROC curve of the data with the Cole frequency as the classifier. The results show that the Cole frequency alone is a viable classifier for malignant breast anomalies. Results of the current work are consistent with recent bioimpedance measurements on single cell and cell suspension breast cell lines.